Volume 32(1); 2024

Optimization of the extracellular secretion of black goat rumen metagenome-derived KG42 xylanase by Bacillus subtilis

Ji-Eun Kim, Jin-Sung Lee, Donghwan Lee, Hoyoon Choi, Keun-Sung Kim

APJMBB 32(1): 1-14

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.01

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Xylanase (E.C. 3.2.1.8) is the enzyme that breaks down β-1,4 xylan by cleaving β-1,4 glycosidic linkages. Production of xylanases is important for various industrial applications. Here, we aimed to determine the optimal incubation conditions for expression and secretion of KG42 xylanase in Bacillus subtilis using response surface methodology based on Box-Behnken design in preparation for industrial applications. Among nine broth media tested in this study, Power Broth was chosen as a basal medium. In addition to the basal medium, the four other independent variables of extra carbon sources (glucose, lactose, mannose, fructose, and sucrose), extra nitrogen sources (beef extract, yeast extract, tryptone, urea, NaNO₃, and (NH₄)₂SO₄), isopropyl β-d-1-thiogalactopyranoside concentrations, and induction times were individually tested using one factor at a time in an optimization experiment. Next, a Box-Behnken design-based response surface methodology approach was used to identify and validate the optimized incubation conditions with the four variables in batch culture. The statistically optimized incubation conditions obtained from this study yielded a maximum of approximately 3- to 4-fold increases in the expression and secretion of KG42 xylanase by B. subtilis in comparison with unoptimized medium and incubation conditions.

Morphological, cytotoxic and apoptotic effects of different concentrations of nisin ZP on MG63 cells in a 3-dimensional culture system

Alyaa R. Alkhateeb, Sharaniza Ab-Rahim, Muhammad F. Azmi, Aisha M. Din, Effat Omar, Gabriele Ruth Anisah Frömming

APJMBB 32(1): 15-23

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.02

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Osteosarcoma (OS) is a primary bone cancer that commonly affects young individuals. Despite treatment, it is known to be highly resistant to therapy. Nisin, a bacteriocin released by Lactococcus lactis has been shown to exhibit an apoptotic effect on cancer cells. To better represent in vivo cancer cell constructs, 3-dimensional (3D) culture systems have been used in vitro. This study aims to compare the effects of nisin on OS cell culture in both monolayered and 3D gas-permeable VECELL^® G-Plate. Osteosarcoma cells (MG63) were cultured on 3D culture (G-Plate) and normal culture plates. Morphological assessments were carried out through light and scanning electron microscopic examinations. The results showed that the use of a 3D system is more reliable in the development of 3D osteosarcoma models compared to the 2-dimensional (2D) culture system. Specifically, the 3D culture demonstrated the formation of MG63 spheroids, indicating the reliability of the hanging drop methods to produce spheroidal cell morphology. Additionally, the cell viability assay showed a higher IC₅₀ value for 2D groups compared to the 3D groups, with values of 102.56 µg/mL and 33.96 µg/mL, respectively. Finally, a dose-dependent apoptosis was noticed for the apoptotic assay, further demonstrating the effectiveness of the 3D culture system. The effectiveness of nisin ZP as an anticancer agent was observed in both 2D and 3D culture systems. However, the 3D culture system required a higher dose of nisin ZP to demonstrate its potential, indicating that this system could be a promising approach in OS intervention studies.

The miR-221/222 regulates the ID1 gene expression through PTEN, c-JUN and ARF4 mediators to control cell proliferation

Panus Yingjamsiri and Saowakon Paca-uccaralertkun

APJMBB 32(1): 24-31

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.03

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Objective: To explore the effect of miR-221/222 on cell proliferation and regulation of inhibitor of differentiation1 (ID1) expression concerning the value of miR-221/22 in cancer diagnosis, prognosis, or therapeutic use. Methods: Embryonic cell line (HEK293), breast cancer cell line (MCF7), and lymphoblast cell line (U937) were employed to investigate cell proliferation in the presence or absence of miR-221/222 using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The relation of miR-221/222 and expression of the ID1 gene was monitored via luciferase activity and real-time PCR. Quantitative PCR was used to detect the effect of miR-221/222 on the expression of cellular proteins, c-JUN, ARF4, and PTEN. Results: The miR-221/222 significantly increased the expression level of the ID1 gene in MCF7 and U937 cell lines but downregulation of the ID1 gene was revealed for transfected HEK293 cells. Level of the ID1 mRNA showed that miR-221/222 regulated the ID1 gene expression at the transcriptional level. Moreover, miR-221/222 enhanced the cellular proteins, ARF4 and c-JUN, and expression in MCF7 and U937, while inhibiting them in HEK293. These findings indicated that the effect of miR-221/222 on ARF4 and c-JUN expression in HEK293 may be mediated by different pathways from MCF7 and U937. Conclusions: Increasing levels of miR-221/222 expression are correlated with cell proliferation and the regulation of ID1 expression. ID1 regulation is a complex process and miR-221/222 may regulate its expression via c-JUN and ARF4. The value of miR-221/22 in cancer diagnosis, prognosis, or therapeutic use may be cancer type dependent.

Multi-locus phylogenies revealed a new record of Entoloma species (Basidiomycota, Agaricales) responsible for gastrointestinal poisoning

Sittiporn Parnmen, Nattakarn Nooron, Ratana Tacharoenmuang, Sujitra Sikaphan, Dutsadee Polputpisatkul, Chutimon Uttawichai, Khwanruan Naksuwankul, Onanong Pringsulaka, Sutheewan Binchai, Achariya Rangsiruji

APJMBB 32(1): 32-43

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.04

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Objective: The frequency of mushroom poisoning has increased worldwide. In Thailand, reports on mycetism between 2013 and 2021 included 18 cases of Entoloma poisoning. Incorrect identification of toxic Entoloma species occurs during wild mushroom foraging, and their consumption results in gastrointestinal irritation. Morphological distinction of mushroom leftovers is difficult; however, we aimed to characterize 40 clinical samples to the genus level using macroscopic and microscopic analyses. Furthermore, species identification of the poisonous Entoloma was performed based on multi-locus phylogenetic analyses of the nuclear internal transcribed spacer region, nuclear large subunit ribosomal DNA, and mitochondrial small subunit ribosomal DNA. Phylogenetic trees reconstructed using maximum likelihood and Bayesian methods demonstrated that forty samples of mushroom remnants formed eight robust clades with six identified species. The findings confirmed the taxonomic identity of E. griseolazulinum as a new record of species causing gastrointestinal poisoning. The use of molecular annotation provided a foundation for the development of accurate methods for identification of toxic mushrooms.

Unlocking the potential of chitosan nanoparticle as a carrier for systemic acquired resistance (SAR)-inducing recombinant protein: A preliminary study

Nur Balqis Zamri, Nur Sabrina Wahid, Norliza Abu Bakar, Mohd Zulfadli Sohaime, Noor Azlina Masdor, Nor Suzaida Mohd Nor

APJMBB 32(1): 44-56

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.05

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Objective: Dieback disease stemmed from a phytopathogenic bacteria named Erwinia mallotivora is disastrous to papaya plant. Disease management action plan occurred stagnantly across the continent, until the discovery of proteinaceous hairpin which may catalyse systemic-acquired resistance (SAR) as plants’ defence mechanism. However, hairpin utilization is seriously flawed by its instability and limited bioavailability in plant. In lieu of this matter, nanobiotechnology approach through the encapsulation of hairpin within chitosan nanoparticles may be a mediator towards enhancement of sustained delivery as well as synergistic effect during foliar application. In this preliminary work, hrpN, a hairpin functional as SAR elicitor from Erwinia mallotivora, was well-expressed and purified in Escherichia coli system at molecular weight and concentration; 30 kDa and 1 µg/µL, respectively. Optimization of ionic gelation simultaneously revealed that optimal chitosan (CS) to sodium tripolyphosphate (TPP) volume ratio was found to be 2.4:1 ratio, yielding nano-sized particles with mean hydrodynamic diameter 66.27 ± 1.77 nm, and homogenously distributed with polydispersity index 0.189 ± 0.027. Subsequently, encapsulation of purified hrpN within CNP was formulated at various hrpN concentration. Following that, encapsulation of 0.04 mg/mL hrpN within optimized CNP produced hrpN-encapsulated chitosan nanoparticles (CNP-hrpN) with small size (106.34 ± 2.053 nm), stable and well-dispersed (0.188 ± 0.011), as well as possessing excellent encapsulation efficiency (81.84 ± 3.43%). The outcome from current work portrayed the potential of chitosan nanoparticle to carry biomolecules with desired properties.

Identification of single nucleotide polymorphisms (SNPs) in selected rice phosphate transporter (OsPHT) genes

Chui Yao Teh , Rattanak Sambath Lee, Kamariah Hasan, Clement Kiing Fook Wong

APJMBB 32(1): 57-64

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.06

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Phosphorus (P) is one of the fundamental elements for plant growth and development. Due to the scarcity of viable P in the soil for plants, P deficiency was often the culprit that restrained plant’s wellbeing. Plasma membrane phosphate transporters (PHT) are a group of proteins responsible for phosphate (Pi) uptake from soil and further allocation to plant organs and tissues. The PHT can be further categorized into constitutively expressed low-affinity Pi transporter or high-affinity Pi transporter that are induced upon Pi starvation. Significant variability in P use efficiency has been observed among different rice varieties. Genotypic differences such as single nucleotide polymorphisms (SNPs) could be responsible for the variation observed aside from the well-studied phenotypic responses. Nevertheless, the occurrence of the SNPs in OsPHT genes remain unexplored. Therefore, the objective of this study was to analyse and profile the SNPs in five selected high affinity OsPHT genes which are responsible for P uptake under P deficiency. The SNPs mining was conducted using Rice SNP-Seek Database against 3024 rice varieties with Oryza sativa japonica cultivar Nipponbare as the reference sequence. Results showed that a total of zero, seven, three, one and ten non-synonymous SNPs was identified in OsPHT1;2, OsPHT1;3, OsPHT1;6, OsPHT1;9 and OsPHT1;10, respectively. A base substitution of C to A at position 16028497 of chromosome 10 of OsPHT1;3 was found to change tyrosine to a stop codon. This could result in a truncated protein which has only 213 amino acids as compared 526 amino acids in the complete protein. The large number of non-synonymous SNPs in OsPHT1;10 could explain the redundant function of this gene in the translocation and uptake of P in rice. In short, the identified SNPs especially the non-synonymous SNPs could potentially disrupt the biosynthesis of phosphate in rice which requires further investigation.

Optimation of the cell density, biomass production, lipid, and carbohydrate content of Nannochloropsis oculata with fluorescein and pH manipulation

Jody Ashrib Satriayudistira, Eko Agus Suyono, Arief Budiman

APJMBB 32(1): 65-75

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.07

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Nannochloropsis oculata is a potential marine microalga. However, the mass production of N. oculata encounters several challenges, such as the high costs of resources, contamination, and light absorption optimization. These problems can be solved by pH manipulation and fluorescein induction on culture media. Various studies have shown that pH values manipulation can cause fluctuations in cell density, dry biomass, and carbohydrate and lipid production of microalgae. On the other hand, fluorescein can act as a molecular antenna that improves light absorption. This research is novel because of its focus on its unique attempt to utilize pH manipulation combined with fluorescein induction to enhance the growth and metabolite production of N. oculata. The implications of this research may provide cost-effective, viable, and sustainable development of the algal industry in general. In this research, cultivation was carried out for seven days with four levels of pH (7 – 10) combined with three variations of fluorescein addition (0, 0.15, and 0.3 mL). Every treatment combination had three replications. Cell density and biomass as growth analysis parameters were calculated during the cultivation using the Haemocytometer and gravimetric methods. Bligh & Dyer and phenol sulfate methods were used to analyze lipid and carbohydrate content. Based on the result, the best treatment combination, pH 9 with 0.15 mL of fluorescein, increases cell density and carbohydrate content of N. oculata by 31.45 and 4.4%, albeit not statistically significant.

Antimicrobial susceptibility of bacterial clinical specimens isolated from Al-Sader Teaching Hospital in Basra-Iraq

Ahmed Mshari Thari, Khairallah A. S. Mohammed, Najwa M. J. Abu-Mejdad

APJMBB 32(1): 76-84

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.08

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Antibiotic resistance is a global health concern that requires multiple efforts to establish proper preventive and curative programs. This study aimed to assess the antibiotic susceptibility profiles of commonly isolated bacteria at Al-Sader Teaching Hospital, Basra-Iraq.  A total of 234 clinical specimens were collected from urine, skin infections, and blood. The bacterial isolates were initially identified using standard microbiological methods, and the identification of the four most common isolates was confirmed by PCR technique using species-specific primers (malB for Escherichia coli, rpoB for Klebsiella pneumonia, OprL for Pseudomonas aeruginosa, and mecA for methicillin resistant Staphylococcus aureus).  The identified bacteria were subjected to disc diffusion and VITEK2 system to test the antibiotic susceptibility. E. coli was the most prevalent in urine (51.25%), followed by S. aureus (15%), while S. aureus (31.25%) and P. aeruginosa (30.20%) were more prevalent in skin infections. S. aureus (31.57%) and E. coli (26.31%) were predominant in the blood samples. The Antibiotic susceptibility pattern of Gram-negative isolates revealed high resistance to Cefoxitin (90%), Aztreonam (87%), Ceftriaxone (87%), Piperacillin/tazobactam (85%), and Amoxicillin/clavulanic acid (85%). S. aureus showed high resistance to Cefoxitin (93%), Oxacillin (89.58), and Methicillin (91%), whereas, the highest sensitivity was recorded for Colistin (100%), Rifampin (93%), Amikacin (87%), and Meropenem (76%). A high level of multidrug-resistance strains was detected among Gram negative bacteria (76%) and S. aureus (56%). The current study provides valuable insights into the distribution of pathogenic bacteria and their antibiotic resistance patterns in different clinical samples, contributing to improved treatment strategies and infection control measures.

Molecular identification of endophytic bacterium DBA2 isolated from the leaf of binahong (Anredera cordifolia (Ten.) Steenis) and its antagonistic activity against bacteria associated with dental caries

Etin Diah Permanasari , Muhammad Ibadurrohman, Susilo Susilo

APJMBB 32(1): 85-92

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.09

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Binahong plants (Anredera cordifolia (Ten.) Steenis) are known to contain secondary metabolites which have important biological properties. The current study focused on the endophytic bacteria which lives in the leaves of A. cordifolia (Ten.) Steenis. The aim of this study is to isolate and identify the endophytic bacteria which can produce antibacterial metabolites from the leaves of A. cordifolia (Ten.) Steenis. Two isolates, DBA1 and DBA2 were isolated and purified from the leaves of A. cordifolia. These isolates were subjected to the screening for their antagonistic activity against the bacteria associated with dental caries, which are Streptococcus mutans and Lactobacillus acidophilus using the disk-diffusion method. The strain of DBA2 exhibited the largest diameter of inhibition zone against both S. mutans (31,17 mm) and L. acidophilus (35,57 mm). While DBA1 exhibited the diameter of inhibition zone of 23,47 mm and 25,87 mm against S. mutans and L. acidophilus, as respectively. The strain of DBA2 was then subjected for molecular identification. The genomic DNA of DBA2 was extracted with the Geno Plus™ Genomic DNA Extraction Miniprep System and molecular identification was performed by PCR amplification and sequencing of the 16S rRNA gene. The amplicons were then purified and sequenced, before the 16S rRNA gene sequences were analysed by a Basic Local Alignment Search Tool (BLAST) search against the National Centre Biotechnology Information (NCBI) database. The endophytic bacterial strain DBA2 from the leaves of A. cordifolia was identified to be closely related to Bacillus sp., and the top match from the database search revealed a similarity value of 100% with the reference Bacillus sp. strain x20. Future studies are required to analyse the bioactive compounds of strain DBA2, which can be considered as a potential source for the new antibacterial drugs for the dental caries treatment.

PSCA gene expression in bladder, colorectal and prostate cancer patients from Basrah governorate southern of Iraq

Anwar Noori Ayoob, Adnan Issa Al-Badran, Rafid Adil Abood

APJMBB 32(1): 93-100

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.10

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Cancer causes death in all countries of the world, with approximately (9.6) million deaths in 2018. Developing countries have high cancer rates. Furthermore, there are risk factors that lead to cancer, including economic and political instability and bad lifestyles. This is the first study of PSCA gene expression with BC, CRC and PC in Iraq, especially in Basrah governorate. This paper involved one hundred and one blood samples being collected from bladder, colorectal and prostate cancer. On the other hand, one hundred and one blood samples with no cancer were collected as a control group. Two ml of peripheral blood was drawn for RNA extraction, then total RNA was reverse‑transcribed to cDNA, and PSCA mRNA was measured using qPCR. The results showed PSCA gene was over expressed in bladder cancer (BC) with an expression level at ± 8.63 for patients and controls estimated with ±4.16. The gene expression was a 2-fold change in patients compared to healthy control. While the PSCA gene was over-expressed in colorectal cancer (CRC) with an expression level ± 8.16 for patients and controls estimated at ±3.30. That means the gene expression was a 2.47-fold change in patients compared to healthy control. The PSCA gene was over expressed in prostate cancer (PC) with an expression level ± 9.47 for patients and controls estimated at ±4.22. The gene expression was a 2.24-fold change in patients compared to healthy control. In the present study, PSCA gene was expressed at a significantly higher level in the BC, CRC and PC patients compared in the controls group.

Exploring actinobacteria isolated from coral originated from Tulamben Bali in inhibiting multidrug resistance bacteria

Fajar Hidayaturohman, Aninditia Sabdaningsih , Diah Ayuningrum

APJMBB 32(1): 101-115

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.11

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Increasing bacterial resistance has reduced the effectiveness of most clinical antibiotics. The limited option of antibiotics to treat Multi-Drug Resistance pathogen infections indicates that there is an urgency to explore new antibiotic candidates. Coral is recognized to produce bioactive compounds that have the potential to develop new medicinal materials with antibacterial properties. The use of marine invertebrates for compound production will reduce populations in nature, therefore associated microbes are preferred due to their environmental friendliness. Actinobacteria is one of the coral-associated microbes capable of producing active compounds of secondary metabolites as antibacterials. This study aimed to identify coral and obtain isolates of Actinobacteria associated with coral potential as an antibacterial against MDR bacteria. This research uses an explorative method. The method used is coral identification based on the morphology and shape of the sclerite. Screening of antibacterial activity was carried out using the agar plug method, microscopic observation using Gram staining and molecular identification of potential isolates by PCR. The results showed that the coral genera obtained were Siphonogorgia sp., Menella sp., Echinomuricea sp., Dendronephthya sp., and Epizoanthus sp. The results of antibacterial activity indicated that 19 isolates inhibited the growth of MDR pathogen bacteria. However, only the potential isolates were identified. The microscopic observations showed isolate III.ISP2.10^-2.3 and III.ISP2.10^-2.6 have Gram-positive coccus and bacillus-shaped, respectively. Potential isolates III.ISP2.10^-2.3 (Micrococcus yunnanensis) were found to have antibacterial activity against Acinetobacter baumanii, Pseudomonas aeruginosa, and Bacillus subtilis, while isolate III.ISP2.10^-2.6 (Brevibacterium pigmentatum) had the greatest activity against B. subtilis.

Isolation and characterisation of potential probiotic yeast strains from local fermented foods: Gastrointestinal tolerance and antimicrobial activity assessment

Amirul Syafiq Murad, Nur ‘Ain Zuhairi, Mohd Amir Shahlan Mohd-Aspar, Mohd Akmal Azhar

APJMBB 32(1): 116-125

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.1.12

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Probiotic microorganisms, offering health benefits when consumed in sufficient quantities, are gaining recognition for their potential role in promoting wellness. This study focuses on isolating and characterising potential probiotic yeast strains sourced from fermented food products. This research evaluates the gastrointestinal tolerance and antimicrobial activity of isolated yeast strains, with the potential application in probiotic supplements and functional foods. Yeast strains were isolated from fermented food sources and identified using morphological analysis, PCR, gene sequencing, and genetic identification. Gastrointestinal tolerance was assessed through simulated gastric fluid (SGF) exposure, and antimicrobial activity was tested against foodborne pathogens. Six yeast strains (Diutina mesorugosa, Pichia kudriavzevii, Candida mesorugosa, Candida sp) were identified. They exhibited varying resistance to low pH in SGF, suggesting survivability in the stomach. Some strains selectively inhibited specific Gram-negative pathogens like Pseudomonas aeruginosa and Salmonella sp. These findings suggest the isolated yeast strains may serve as probiotics, promoting digestive health and food safety. They are potentially used in probiotic supplements and functional foods, promising improved overall well-being.

Volume 32(2); 2024

Bacterial nanocellulose (BNC) biosynthesis by Komagataeibacter hansenii RM-03 using agricultural waste as substrates and BNC-silver nanocomposite preparation

Aini Darwina Daud, Nor’Aini Abdul Rahman, Hooi Ling Foo and Rosfarizan Mohamad

APJMBB 32(2): 1-14

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.01

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Bacterial nanocellulose (BNC) is a remarkable biopolymer synthesised by bacterium, exhibiting exceptional properties. However, conventional Hestrin-Schramm (HS) medium, particularly the carbon source, poses challenges of high costs and low productivity. This study explores BNC biosynthesis on a modified HS medium, employing agricultural wastes (sugarcane molasses, banana peel, and pineapple peel) as carbon sources, and compares the overall yield of BNC produced. Sugarcane molasses proved to be the most effective, yielding the highest BNC concentration (8.19 g/L) after 7 days, followed by pineapple peel (2.16 g/L) and banana peel (2.11 g/L). Extensive research was conducted to enhance properties of BNC by an environmentally friendly approach, incorporating silver nanoparticles (AgNP) utilising Momordica charantia fruit extract, resulting in a BNC-Ag nanocomposite. The synthesis involved mixing 1 mM silver nitrate (AgNO₃) with 15 mL of M. charantia fruit extract to reduce Ag ions to AgNP, which was confirmed by UV-vis spectroscopy with an absorbance peak between 400 and 410 nm. Characterisation using FESEM and TEM on the synthesized BNC showed minimal impact on BNC fiber diameter from waste-derived carbon sources. XRD indicated slight variations in crystallinity index, with the highest (85%) in TSM-derived BNC. FTIR analysis revealed similar chemical profiles across all BNC, indicating cellulose formation. The BNC-Ag nanocomposite exhibited potent antibacterial activity against multi-drug resistant strains (Pseudomonas aeruginosa, Salmonella typhi, Bacillus subtilis, Staphylococcus aureus) through disc diffusion method with inhibition zones up to 16.8 mm. Overall, the findings from this study contribute to the development of environmentally sustainable for the production of functional BNC materials with enhanced properties for diverse applications.

Comparison of CO₂ absorption via terrestrial plants and microalgae: A review

Tia Erfianti, Istini Nurafifah, Brilian Ryan Sadewo, Budi Setiadi Daryono, Eko Agus Suyono and Arief Budiman

APJMBB 32(2): 15-26

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.02

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Currently, global warming and climate change continue to increase along with CO₂ gas emissions. This has an impact on the survival of organisms, including humans. Therefore, efforts to reduce CO₂ emissions have been conducted by various methods, such as chemical, physical and biological methods, one of the most efficient methods to absorb CO₂ gas is to use microalgae. Microalgae are photosynthetic organisms capable of absorbing CO₂. Microalgae can also be converted into valuable products such as biofuels, biofertilizers, food, feed, medicines, and cosmetics through an integrated biorefinery concept. In the future, CO₂ mitigation using microalgae will be massively studied, considering the many benefits obtained from the utilization of microalgae to reduce CO₂ emissions in the world. Through the concept of biorefinery, microalgae can be processed into various derivative products that are useful for humans in the food, feed, health, industrial, medicine, and cosmetic sectors. This review will compare the effectiveness of CO₂ absorption through terrestrial plants, microalgae, and microalgae-bacterial consortia to the possibility of its application and challenges.

Tetraspanin CD9 peptides for membrane disruptive on P. aeruginosa

Khairiyah Murad, Sharaniza Ab-Rahim and Hassanain Al-Talib

APJMBB 32(2): 27-30

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.03

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The rapid development of multidrug-resistant strains and nosocomial P. aeruginosa infections pose a growing threat in the clinical setting. Tetraspanin CD9 peptides has demonstrated the antimicrobial activity against various gram-positive and -negative bacteria. Recently, CD9 peptides have shown anti-adhesion properties against P. aeruginosa isolates. In this study, we investigated the effect of CD9 peptides on the membrane of P. aeruginosa by transmission electron microscopy (TEM). The result shows the CD9 peptides cause disruption of the membrane of P. aeruginosa. In addition to the established antibacterial properties, this work reveals that tetraspanin CD9 peptides, a membrane-disrupting drug, could possibly be used as an additional treatment approach against P. aeruginosa. Future research should incorporate confocal microscopy to locate the bacterial matrix components and distinguish between living and dead P. aeruginosa upon the treatment with CD9 peptides.

Copper surface acts as good surface for biofilm attachment

Srinivasa Sundara Rajan R

APJMBB 32(2): 31-37

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.04

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The drinking water distribution system is a huge network of pipes that travel miles before reaching the household. The age of the pipe and the material affect the water quality. The water monitoring and testing parameters are limited to water quality and not pipe health. The pipe material highly impacts the biofilm formation.  Copper is commonly used for its anti-microbial property, but also aids for the attachment of bacteria onto the surface. This study was aimed at analyzing the formation of biofilm on copper in a drinking water. Copper coupons were immersed in a drinking water system for a period of six months and analyzed for biofilm formation. Microbiological enumeration were done by Heterotopic plate count and found an average of 35 CFU/ml in water and 50 CFU/ml on the copper coupon. The average copper content in water was 0.01 mg/ml. SEM analysis revealed the formation of bacterial growth and found that cocci shaped bacteria were predominating on the surface of the coupons. The copper coupon serves as a surface for the attachment of bacteria and aids in the formation of biofilm.

Morphological analysis, agronomic characteristics, species identification, and phylogenetic analysis of Pseuderanthemum sp. using the rbcLgene

Thieu Van Duong, Do Van Mai, Pham Thanh Trong, Tran Van Be Nam and Mai Van Hung

APJMBB 32(2): 38-44

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.05

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The study aimed to evaluate the agronomic characteristics of eleven samples of Pseuderanthemum sp. variety based on the results of the morphological survey on stem height, root length, leaves, combining molecular biology methods to determine their genetic relationships by building a phylogenetic tree based on the plastid gene rbcL sequence. Initial results of an evaluation of the genetic diversity of eleven Pseuderanthemum sp. samples showed that there were differences in stem length, leaf width to root length. Based on the analysis results from the pedigree tree, the tree samples with similar survival indexes and genetically close to each other were classified into two main groups. Based on molecular data, eleven varieties of Pseuderanthemum sp. in the study have been identified, all belonging to the species Pseuderanthemum sp. have a high similarity coefficient above 98% and 100% coverage. Although the geographical location of the samples is variable, it is necessary to investigate other gene sequences related to morphological and biochemical characteristics, such as bioactive compounds in leaves and fruits to have more accurate conclusions about this species.

Microplastics abundance in sea cucumber Holothuria scabra from Pulau Malawali, Sabah, Malaysia

Sarah Syazwani Shukhairi, Nurzafirah Mazlan , Nur Nashrah Abd Rahman, Muhammad Nor Afdall Nazahuddin, Amir Syazwan Shawel, Audrey Daning Tuzan, Nurul Ain Jais, Vijay Subbiah Kumar, Safaa Najah Saud and Atna Permana

APJMBB 32(2): 45-56

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.06

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The growing amount of plastic waste in the environment is a result of the increased use of plastics in numerous areas of our daily lives. These plastics are broken down into microplastics (MPs), which are 5 mm or smaller in size. Sea cucumbers are consumed as an exotic delicacy worldwide, however, it is suspected that they could ingest the microplastics due to their nature as filter feeder. In this study, we have identified the MPs presence in the intestines of sea cucumber Holothuria scabra. A total of 30 sea cucumber H. scabra were collected during low tide at a muddy sandy area at Pulau Malawali using bare hands. The gastrointestinal tracts of H. scabra were removed and digested in NaOH. MPs were identified using light microscope and categorized into shapes and colours. Micro-Fourier Transform-Infrared Spectroscopy (µFTIR) analysis was carried out to identify the types of polymers. The majority colours of MPs were black (69.84%) and blue (18.65%). Fibres (98.38%) made up the majority of MPs followed by fragments (1.37%) and films (0.25%). Ethylene Vinyl Acetate (EVA), Melamine-Formaldehyde (MF) and Rayon were among the polymers identified. In conclusion, MPs were found in the intestines of the sea cucumber H. scabra and this could be from the ingestion of polluted sea bed. The importance of studying the presence of these MPs in sea cucumber allows us to assess the impact of plastic pollution on marine animals and ecosystems and understand the potential risks to both sea cucumber and human as consumers.

Tocotrienol decreases β-amyloid mediated toxicity in Caenorhabditis elegans model of Alzheimer's disease

Yuen Chee Wah, Mardani Abdul Halim , Vikneswaran Murugaiyah, Nazalan Najimudin, Ghows Azzam

APJMBB 32(2): 57-65

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.07

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Alzheimer's disease (AD) is a neurological condition that arises from the buildup of senile plaques, which consist of β-amyloid peptide (Aβ), in the brain. The disorder hampers cognitive functions and deteriorates gradually as time passes. The objective of this investigation is to evaluate the efficacy of tocotrienols as a therapeutic intervention for Alzheimer's disease. The study employed a genetically modified Caenorhabditis elegans model that displays paralysis as a result of the activation of the human beta-amyloid Aβ42 gene. This investigation aimed to explore the potential of blended tocotrienols in alleviating paralysis symptoms. Four experiments were conducted to observe the rescuing effect of tocotrienols: paralysis assay, measurement of ROS, immune-dot blot assay, and aggregation assay.  The composition of the mixed tocotrienols consisted of 12.1% α-, 2.7% β-, 18.6% γ-, and 8.1% δ-tocotrienols. Treatment with mixed tocotrienols at concentrations of 0.5 %, 0.75 %, and 1 % noticeably prolonged the onset of Aβ-induced paralysis in the genetically modified nematode. Additionally, tocotrienols exhibited antioxidant properties against Aβ-generated oxidative stress, with 0.1% to 1% of mixed tocotrienols demonstrating significantly reduced ROS in the worms. Furthermore, the combination also exhibited strong inhibitory effects on the aggregation of Aβ, demonstrating a potent activity with an IC50 value of 600 ng/ml.

Withania somnifera showed neuroprotective effect and increased longevity in Drosophila Alzheimer’s disease model

Mardani Abdul Halim , Izzah Madihah Rosli, Siti Shafika Muhamad Jaafar, Ooi Hui Min, Leong Pui Wei, Nurlina Rosli, Shaharum Shamsuddin, Nazalan Najimudin, Ghows Azzam

APJMBB 32(2): 66-70

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.08

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Alzheimer’s disease is a unique neurodegenerative condition in humans that is characterized by amyloid beta accumulation in the brain parenchyma. Withania somnifera, commonly known as Ashwagandha, is a plant that has been used for centuries in Indian Ayurvedic medicine to treat various health conditions. The active compound in Ashwagandha has shown to be beneficial in treating several neurodegenerative diseases, including Alzheimer's disease (AD). This study used a Drosophila melanogaster AD model to investigate the effect of Ashwagandha in reducing amyloid beta toxicity and promoting longevity. The findings showed that 20 mg/mL of Ashwagandha root powder effectively rescued the "rough eye phenotype" in AD Drosophila and increased longevity in both AD model and wild-type Drosophila. Overall, the results suggest that Ashwagandha may have potent therapeutic potential for treating AD and maintaining cellular well-being.

Genomic surveillance and sequencing of SARS-CoV-2 in Malaysia

Norazimah Tajudin, Seok Mui Wang, Darshan Chandra Kumar, Mariam Mohamad, Siti Farah Alwani Mohd Nawi

APJMBB 32(2): 71-83

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.09

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This manuscript offers an in-depth review of the genomic surveillance of SARS-CoV-2 variants in Malaysia, emphasizing the integral role of this surveillance in understanding the virus's evolution and informing public health responses. Leveraging platforms like GISAID, Nextstrain, and the Pangolin classification system, researchers in Malaysia and their global counterparts share genome sequences and clinical data of SARS-CoV-2. These tools, particularly Nextstrain for real-time tracking and visualization of viral evolution, and Pangolin for lineage classification have advanced understanding of significant mutations, such as D614G and N501Y, and their impact on virus transmissibility and pathogenicity. The study of the virus's emergence in Malaysia offers vital insights into its evolutionary trajectories, aiding in effective pandemic management. Malaysia's Genomic Surveillance Program, aligned with national immunization efforts, plays a key role in identifying and controlling COVID-19 spread. This program integrates molecular, epidemiological, and clinical data that helps the health authorities in making a decision that leads to public health intervention and policymaking. This review details the significance of genomic surveillance in Malaysia, emphasizing its impact on understanding viral evolution, monitoring variants, informing public health strategies and responses, and preparing for future infectious disease challenges effectively.

Review and bibliometric analysis of AI-driven advancements in healthcare

Yi Jie Wang, Wei Chong Choo, Keng Yap Ng

APJMBB 32(2): 84-97

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.10

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Purpose: This research intends to use literature review and bibliometric analysis methods to visually review the development status and important historical milestones of Artificial Intelligence, as well as the basic research, key topics, and future potential research hot spots of AI in the healthcare field. Methodology: Conduct in-depth analysis of AI in healthcare through bibliometrics methods such as publication activity analysis, co-occurrence analysis, and co-authorship analysis. Findings: This study outlines the development time trajectory of AI technology and its application in healthcare. Research shows that "algorithm", "machine learning", "deep learning", "controlled study", "major clinical study" and "healthcare delivery" as well as "decision support systems" are key topics for research. Gender-related research and ethical issues are areas of future focus. Research implications: The practical significance is that it can clarify and optimize the key directions of AI to improve the quality of medical decision-making, improve diagnostic accuracy and guide market investment. The originality is reflected in the comprehensive analysis of the development trajectory of AI in the medical and health field. Through a unique perspective and systematic approach, it provides an important reference for research trends and future directions in the field.

Nitrofurazone biodegradation kinetics by batch fermentation of Aspergillus tamarii

Nur Aisyah Mohamad, Muhammad Zafri Zamri, Muhammad Naziz Saat, Zaidah Zainal Ariffin

APJMBB 32(2): 98-109

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.11

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Nitrofurazone (NFZ) compound contains a 5-nitrofuran ring structure that has been widely used as feed additives in animal husbandry. Due to the benzene ring structure, the residues are highly toxic to humans and animals. A kinetics study of NFZ biodegradation in batch fermentation with Aspergillus tamarii KX610719.1 was conducted. The main objectives were to determine the kinetic parameters of fungal growth, glucose consumption, protein production, and biodegradation of NFZ using fungal biomass. Kinetic parameters were determined using Polymath 6.0 software, and regression analysis was done using linear and non-linear methods. After 168 hours of batch fermentation, the maximum specific growth rate (µ_max), and maximum cell concentration (X_max) for cultivation without NFZ were 0.062 h^-1_, and 0.529 g L^-1, respectively. The maximum specific growth rate (µ_max), and maximum cell concentration (X_max) for cultivation with NFZ were 0.092 h^-1_, and 0.327 g L^-1, respectively. For glucose consumption, kinetic parameters of Yield of biomass over the substrate (Y_XS) and cell maintenance (m_S) were estimated at 0.139 g g^-1 and 0.239 h^-1, respectively. Based on the Luedeking Piret model, the estimated growth-associated (α) and non-growth-associated (β) constants were 1.142×10^-2 g g^-1 and 5.680×10^-5 h^-1, respectively. The rate constant (k₁) of NFZ biodegradation was estimated at 2.696×10^-2 h^-1 following the first-order model where the rate constant of NFZ removal is dependent on the NFZ concentration. The application of A. tamarii batch fermentation in the removal of NFZ compound was sufficient with a total percentage removal of 85.9 % or 0.430 g L^-1 recorded.

Investigation of biomimetic HAp formation on graphite

Reem Saadi Khaleel, Mustafa Shakir Hashim , Murthada Kutheir Abbas

APJMBB 32(2): 110-116

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.12

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According to reports, coating implants with hydroxyapatite (HAp) promotes bone repair and combines the biological effectiveness of the material with the mechanical properties of standard metallic implants. In this contribution, bioactive nanographite particles were synthesized using a novel technique to stimulate HAp deposition using a biomimetic method. The rapid breakdown ionization (RBA) technique was used to synthesize graphite nanoparticles. The electrophoretic deposition (EPD) technique was utilized to deposit the nanoparticle on titanium substrates. Scanning electron microscope (SEM) images showed the creation of nanoparticles with a size of around 65nm. An X-ray diffraction (XRD) test confirmed the polycrystalline structure of graphite with a dominant peak (002). UV-VIS absorption and FTIR spectra confirmed the production of graphite powder. To test the bioactivity of the graphite layer, it was immersed in simulated body fluid (SBF) for 30 days. The formation of a HAp layer on graphite is depicted by an XRD pattern, and SEM images illustrate nanoclusters of this layer.

Effect of storage condition on the viability of sago effluents as carbon source in fermentation medium for bioethanol production

Mohamad Zulhisyam Rashid, Dyg Salwani Awg Adeni , Muhammad Norhelmi Ahmad

APJMBB 32(2): 117-126

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.13

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In Sarawak, Malaysia, approximately 237 tons/day of sago effluent is commonly discharged into nearby river due to the sago starch extraction process. Due to the high concentration of polymeric compounds, particularly starch, in sago wastewater, which petrifies easily, this condition severely pollutes the environment in the affected area. This study was conducted to determine the viability of using sago effluent as a carbon source and fermentation medium for bioethanol production which indirectly help to minimize the environmental impact as well as the economics of the sago industry. The sago effluent obtained from the local sago mill was analysed for starch content and pH profile while stored at room and cold (4°C) temperature facility. Enzymatic hydrolysis was conducted to convert the residual starch into glucose as carbon source for bioethanol fermentation. Fresh sago effluent can be stored for up to 5 days in cold temperature where the starch content remains constant. The highest starch concentration in sago effluent was 61.33 g/L, in which 50.57 g/L glucose was obtained through the enzymatic hydrolysis process. Hence 82.5% of the starch to glucose conversion yield is revealed. Then, the sago effluent hydrolysate which acts as a carbon source as well as a fermentation medium able to generate 23.14 g/L of bioethanol, displays a 91% theoretical yield of glucose to ethanol. In conclusion, the utilization of sago wastewater as feasible alternative to cheap and locally available and sustainable source of raw materials to produce bioethanol.

Exploring the antibacterial potential of tunicate-associated bacteria (Ascidiacea) at the shipwreck site of Menjangan Kecil Waters, Karimunjawa

Aen Hendrawati, Diah Ayuningrum , Aninditia Sabdaningsih, Rosa Amalia

APJMBB 32(2): 127-136

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.14

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Menjangan Kecil Island is situated south of Karimunjawa Island and boasts a rich diversity of coral reefs and associated marine life, including tunicates. Tunicates, also known as sea squirts, produce secondary metabolites to defend against predators. This study aims to explore the antibacterial potential of tunicate-associated bacteria from shipwrecks in Menjangan Kecil Waters. The research adopts an exploratory descriptive approach. Seventy-two bacterial isolates were recovered from five tunicate specimens. Fifteen isolates exhibited antibacterial activity against Vibrio harveyi, with two isolates, KJ1.3.02.05 and KJ1.4.02.02, displaying the highest activity. Molecular identification via 16S rRNA gene amplification revealed that KJ1.3.02.05 was Sinomicrobium oceani (99.64% similarity), and KJ1.4.02.02 was Bacillus haynesii (98.64% similarity).

Expression of Heat Shock Protein 70 in thyroid gland tumors

Haider A. Hassan , Saad Hasan Mohammed Ali, Athraa Y. Al-hijazi

APJMBB 32(2): 137-143

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.15

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Heat shock protein 70 (HSP70) is a crucial protein with vital biological tasks in cell continuation of life. The variation of HSP70 activation occurs as a consequence of stress that includes temperature states, toxicity, poisoning with heavy metals, and tumor-related conditions. One of the master jobs of the HSP family is the suppression of caspase-mediated apoptosis signals. A high level of the expression of HSP70 is accountable for tumorigenesis and resistance against chemotherapeutic drugs. For this reason, the detection of HSP70 may help to diagnose cancerous diseases. From the other side, targeting this chaperone might help in treatment by maintaining late caspase-dependent events. This study was conducted to detect the presence and the location of HSP70 in Iraqi thyroid tumor tissue specimens (25 samples), in addition to 10 samples of normal thyroid tissue. Using the immunohistochemical study (paraffin method), the protein was detected in 100% of follicular carcinoma or follicular adenoma (benign) in addition to 77.7 % of papillary thyroid carcinoma while, in normal thyroid tissue, the presence of protein was in 10 % of cases. Regarding protein location in the cells, it appeared in the nuclei and the cytoplasm of follicular carcinoma cases in comparison with just in the cytoplasm of other sections.

Range extension of Malaysian Earth Tiger Tarantula Omothymus schioedtei in Kuala Pilah, Negeri Sembilan, Malaysia.

Aida Syafinaz Mokhtar , Norhidayu Sahimin, Nurliyana Mohamad, Nurhamimah Zainal Abidin, Muhammad Aidil Ibrahim, Muadz Ahmad Mazian, Amirul Adli Abd Aziz

APJMBB 32(2): 144-147

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.16

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With little information on this mygalomorph species apart from its morphological characteristics, we report the first sighting of Omothymus schioedtei in Universiti Teknologi MARA (UiTM) campus located in Kuala Pilah, Negeri Sembilan, marked the southernmost extended range in Peninsular Malaysia for this species. The species identity of this previously unknown theraphosidae was confirmed by DNA barcoding of the mitochondrial marker cytochrome oxidase subunit I (COI). It is anticipated that the COI DNA barcoding technique used for the specimen viewed in this study will be able to provide rapid and accurate identification of spiders.

Potential and challenges of DNA analysis in Natural History Collections

Fatima Bachir, Hajar Darif, Oumnia Himmi, Mohammed Fekhaoui

APJMBB 32(2): 148-159

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.17

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Natural History Collections (NHC) serve as repositories of a wide range of specimens that are valuable resources for genetic studies. These specimens, housed in natural history museums, herbaria, or research institutes, can be accessed physically or, increasingly, online, due to current digitization efforts. They provide insights into past populations unaffected by recent anthropogenic and climatic changes, as well as those that have become extinct. However, the DNA extracted from NHC specimens is often degraded and susceptible to contamination, posing challenges for genetic analysis. PCR-based approaches can be used to amplify short mitochondrial DNA sequences, allowing for rapid species identification. However, studying nuclear loci using PCR can be costly and time-consuming due to the need for multiple amplifications. The advent of High-throughput sequencing (HTS) has brought a revolution in NHC-DNA research enabling the sequencing of degraded DNA at a more affordable cost. Different methods, such as whole genome sequencing, sequence capture, and restriction digest, have been employed in HTS studies. Whole genome sequencing provides nuclear and mitochondrial sequences but requires a large amount of high-quality DNA. Reduced genome representation methods, such as sequence capture and restriction digest, help reduce sequencing costs but have technical challenges and limitations. Despite the advantages of HTS, NHC-DNA studies face constraints such as incomplete records and the potential for harm to specimens during DNA extraction. NHC specimens should be preserved for future research, and scientists must carefully balance the benefits of genetic analysis with the conservation of these irreplaceable resources.

Phytochemicals from ginger with anti-viral property as a COVID-19, Mpro inhibitor – An In-Silico study

Sharvari Kulkarni Punde, Pramodkumar P Gupta, Mala M Parab, Sharvari U Devane, Divija M Bhivate, Girija A Bhalkar, Supriya R Chavan, Pallavi S Patil

APJMBB 32(2): 160-168

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.18

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The prompt and globally widespread of a novel Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has created an immediate need to invent new and novel management in the treatment of COVID-19. Thus, this study aims to study the phytochemicals found in ginger that may exhibit anti-viral properties against COVID-19 main protease (Mpro). Here we have screened the compounds having anti-viral properties from ginger deposited in the IMPATT database, screened with ADMET (SWISS-ADME) properties, followed by the molecular docking using the Autodock-Pyrx tool with main protease of SARs-COV-2. The optimum dock complex was studied in MD simulation using Desmond for the dynamic behavior over a period of 100 ns. From the 96 compounds identified in the IMPATT database, followed by ADMET and molecular docking-based screening the compound CID_5282110, Cinnamyl acetate was finally evaluated for the dynamic simulation and reported with optimum outcome. This compound could be further considered for in-vivo and in-vitro-based studies.

Glucose production from wheat straw by cellulase activity produced by local isolate of Aspergillus terreus AJ3 under solid-state fermentation

Ali J. R. AL-Sa'ady, Aida H. Ibrahim, Zainab W. Abdulameer, Sumaya Saady, Mohanad S. Al-Fayyadh, Dunya M. Ahmed

APJMBB 32(2): 169-183

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.19

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Glucose is considered as one of the most important monosaccharides, consisting of six carbon atoms. Glucose can be bound with other sugars or with other glucose to form complex compounds or polysaccharides such as cellulose. Therefore, it is possible to biodegrade cellulose to produce glucose using the cellulase enzyme produced by microorganisms. One of the significant filamentous fungal isolates like Aspergillus terreus can be used for this purpose. Aspergillus terreus AJ3 was activated via culturing on potato dextrose agar media, then the optimum conditions were determined for cellulase and glucose production by using this isolate. The better parameters after investigation were wheat straw, corn step liquor as nitrogen source, moisten at ratio 1:1 (v:w) with mineral salts solution at pH 6.0, and were incubated at 30°C for 6 days. The cellulase purification date demonstrated that, following precipitation by ammonium sulfate (0-75%), gel filtration (Sephadex G-150) was an effective procedure for enzyme purification, with specific activity of around 1433.25 U/mg, yield of approximately 49% and 2.45 as purification fold. The findings of enzyme characterization demonstrated that the molecular weight of cellulase was 26 kDa, and the best pH for cellulase activity was 4.5 and the pH stability was ranged from 4.0-8.5. Additionally, the better temperature for cellulase activity was 40°C, while the thermal constantly of enzyme was ranged from 20-50°C. The Thin Layer Chromatography outcomes for glucose detection showed that the wheat straw and cellulose were hydrolyzed to glucose, depended on retention factor (Rf) values of the standard glucose and the test samples (0.36).

Isolation and characterisation of Pseudomonas aeruginosa bacteriophage isolated from Batu Pahat, Johor, Malaysia

Nur Izzatul Iman Hairil Azmi, Yue-Min Lim, Yap Wei Boon, Muttiah Barathan, Kumutha Malar Vellasamy, Vanitha Mariappan

APJMBB 32(2): 184-192

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.2.20

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The World Health Organization has classified Pseudomonas aeruginosa as a 'Priority One and Critical Pathogen' for which research and design of new antibiotics are urgently needed due to its high rate of antimicrobial resistance. Phage therapy, which uses bacteriophages (phages), has been proposed as an antibacterial agent and shows potential for combating this issue. This study aimed to isolate and characterise bacteriophages from different environmental samples that act specifically against P. aeruginosa. The phages were tested to determine their ability to lyse P. aeruginosa using a spot test. Transmission electron microscopy (TEM) was employed to determine the structure, size and phage family, while specificity and sensitivity tests were conducted using six different bacterial species and 20 clinical multi-drug resistant P. aeruginosa isolates, respectively. Phage PA1 was isolated from Batu Pahat, Johor and using a spot test, PA1 could form clear plaques against P. aeruginosa. PA1 was present in a high titer of 1.06 (± 32.2) x 10¹⁰ PFUs/ml. Based on TEM analysis, PA1 was classified as a member of the Myoviridae family. Host-range analysis displayed that PA1 had 100% specificity towards P. aeruginosa and only 45% sensitivity towards different P. aeruginosa clinical isolates. Phage PA1 demonstrated lysis of P. aeruginosa but exhibited a narrow host range, presenting a challenge for phage therapy. A promising approach to overcome this limitation involves using phage cocktails containing multiple strains of phages to broaden the host range and enhance the overall efficacy of phage therapy.

Volume 32 (2) Supplementary Issue

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Volume 32(3); 2024

Identification of V59L and A953G genotypes distribution in AQP7 and their association with glycerol in overweight/obese Malay patients

Mardhiah Masri, Ruzi Hamimi Razali, Thuhairah Abdul Rahman, Fathimah Mohamad, Xin Wee Chen, Aletza Mohd Ismail, Rohana Abdul Ghani, Rose Adzrianee Adnan, Arjoanna Farra Azizi

APJMBB 32(3): 1-10

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.01

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One significant public health issue contributing to cardiovascular risk factors is obesity. Previous evidence suggested that abnormal glycerol metabolism and aquaporin 7 (AQP7) dysfunction in promoting glycerol influx and efflux from the adipose tissue are among the mechanisms involved in obesity. This pilot study aims to identify the genotype distribution of polymorphisms in the AQP7 gene (AQP7) and to find their association with plasma glycerol. A cross-sectional study was undertaken at Hospital Universiti Teknologi MARA (HUiTM) Sungai Buloh, Selangor, Malaysia, on 56 normal and 44 overweight/obese participants. Anthropometry data was collected from all participants. Blood samples were taken by venipuncture to measure plasma glycerol and subsequently, the genotypes of two SNPs in AQP7 (V59L rs4008659 and A953G rs2989924) were determined for both groups. The genotype distribution and allele frequencies of both SNPs in the AQP7 were established, and their association with plasma glycerol was estimated by logistic regression. Participants in the overweight/obese group had higher plasma glycerol (median = 0.78 mg/dL, Q1-Q3=0.47-1.42) than the normal group.  V59L and A953G genotypes distribution between normal and overweight/obese groups showed no significant difference. Logistic regression analysis showed that participants with the A953G (rs2989924) TC genotype had a 71% decreased risk of developing abnormal plasma glycerol when factors such as age, gender, and waist-to-hip ratio (WHR) were controlled. No similar association was seen in the participants with the V59L (rs4008659) genotypes. This study highlighted the potential role of the A953G (rs2989924) TC genotype in reducing the risk of having impaired glycerol metabolism, reflecting its likely protective nature against obesity.

Association of vitamins and minerals with COVID-19: A mini review

Khaldoon Jasim Mohammed, Ahmed Falah Imran, Salam Kitab Rubat

APJMBB 32(3): 11-20

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.02

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of coronavirus disease 2019 (COVID-19). SARS-CoV-2 has the potential to cause multiorgan failure (MOF), which is lethal. This review aims to evaluate the levels of some vitamins (vitamins C, A, D, and E) and minerals (zinc, iron, and selenium) in individuals infected with coronavirus. All studies reviewed indicate a deficiency in the minerals and vitamins mentioned above. We conclude that micronutrients play a significant role in the immune system and their deficiencies have a significant effect on the immune system. Although most antioxidants are believed to be present in diets, it has been suggested that taking more antioxidants will boost immunity. Although the immune system is constantly active, specific immunity is activated more when pathogens are present. The metabolites of vitamin A, D, and other nutrients control the expression of genes in immune cells.

Immunomodulatory activity of mahanimbine from Murraya koenigii (Curry Leaf) on lipopolysaccharide-induced RAW 264.7 macrophages

Nor Malia Abd Warif , Nur Vaizura Mohamad, Vanitha Mariappan

APJMBB 32(3): 21-27

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.03

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One of the kinds of carbazole alkaloids isolated from Murraya koenigii leaves is mahanimbine. Although this substance has the potential to be an immunotherapeutic agent such as immunostimulatory, antibacterial, antiviral and anticancer, however, its immunomodulatory effects have not yet been completely characterised. Mahanimbine's immunomodulatory effects on RAW 264.7 macrophages induced by lipopolysaccharide (LPS) were investigated in this research. Mahanimbine was applied to the RAW 264.7 macrophage cells for 24 hours, and the cell viability was assessed using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) test. Using an ELISA detection assay, the production of IL-1β and TNF-α by stimulated RAW 264.7 cells was quantified. The substance had no cytotoxicity or effect on cell viability at any of the measured concentrations. In addition, mahanimbine did not demonstrate any changes in nitrite oxide production (14.06-15.40 μM) and IL-1β (251.0–238 pg/ml). However, the compound significantly enhanced the TNF-α production (344.4-426.0 pg/ml). The findings provide evidence of the immunomodulatory potential of mahanimbine by regulating pro-inflammatory cytokines.

Antimicrobial activity of mangrove bark tannins and copper ions on marine bacteria

Sharifah Radziah Mat Nor, Darah Ibrahim, Mohd Jain Kassim, Siti Aisyah Daud, Nor Hawani Salikin, Syarifah Ab Rashid

APJMBB 32(3): 28-38

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.04

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A novel sealant or antifouling agent is desperately needed since numerous antifouling biocides, including tributyltin and diuron, have been banned due to their hazardous effects on the marine environment. Mixed-tannin extracted from Rhizophora apiculata was selected as the natural source to be combined with copper ions to form a copper-tannate (Cu-T) complex. The Cu-T complex was tested for antimicrobial properties via disc diffusion assay against a series of identified marine fouling bacteria. The zone of inhibition obtained from the effect of Cu-T on test bacteria ranged from 10.3 mm-21.0 mm. Meanwhile, the range for minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) varied from 0.25 mg/mL-1.00 mg/mL and 0.50 mg/mL-2.00 mg/mL, respectively. The Cu-T complex affected the growth of eight marine fouling bacteria, with bacteriostatic effect on Bacillus aquimaris IBRL FB13 and Vibrio alginolyticus IBRL FB6. Time kill assay which was performed at the extract concentrations of 1/2MIC, MIC and 2MIC revealed that the antibacterial activity of Cu-T complex was concentration-dependent. From the photomicrographic observation, the Cu-T complex deteriorated the cell walls of the Gram-positive and Gram-negative bacteria. This study highlights the potential of the Cu-T complex as a sealant for mitigating biofouling formation on aquaculture equipment.

Enhancing the performance of Methylorubrum extorquens AAZ-1 by using CRISPR/Cas9 for the degradation of hydrocarbon compounds

Anwar A. Maki, Asaad M. R. Al-Taee , Zeenah Weheed Atwan

APJMBB 32(3): 39-48

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.05

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An efficient expression method was used to create a genetic modification strategy for methylotrophic bacteria to improve the capacity of bioremediating bacteria to absorb oil in contaminated environments. Using the plasmid-treated CRISPR-Cas9 system, Methylorubrum extorquens AAZ-1(OR226417.1) was treated, and qPCR analysis revealed that the expression of the MxaF gene (encoding methanol dehydrogenase enzyme) rose six-fold. To increase the biodegradation effectiveness of hydrocarbons (n-alkane and PAH), CRISPR-Cas9 technology was selected for the investigation and use in the laboratory. The biodegradation rate of n-alkane in the modified M. extorquens AAZ-1 rose from 61.14% to 74.35% during a seven-day incubation period when compared to the control. The proportion of polycyclic aromatic hydrocarbons in the altered M. extorquens AAZ-1 rose from 65.69% to 78.23%. To the best of the authors’ knowledge, this work is the first time that a CRISPR-Cas9 system has been employed to improve the efficiency of these bacteria in the biodegradation of hydrocarbon molecules.

Iraqi patients with a single-nucleotide polymorphism of interleukin-10 -1082G/A and interleukin-6 -174G/C susceptibility to asthma

Rana Talib Mohsen, Al-Taee Haneen Z, Wafaa Hussien Habeeb, Anmar Kamil Alalwani

APJMBB 32(3): 49-55

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.06

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The effects of genetic variations in the IL-10 -1082G/A gene and IL-6 -174G/C gene, as well as the genotypes and alleles linked to the prevalence of asthma disease, were investigated using a molecular and immunological study. Between October 2018 and the end of July 2020, 40 healthy individuals (20 females and 20 males) served as a control group for the study, which involved 50 asthmatic patients (31 females and 19 males) at the Allergy Centre, Al-Anbar Teaching Hospital, in Al-Anbar City. The study used the Single Specific Primer-Polymerase Chain Reaction (SSP-PCR) technique to show that the single nucleotide polymorphisms IL-10 -1082G/A and IL-6 -174G/C had a considerably high prevalence rate (P<0.05) among asthma case and that there was an association between the polymorphism and the asthma risk. The findings indicate that asthma patients had considerably higher (P<0.05) IL-10 A alleles and heterozygous GA genotypes (1082G/A) compared to the control group. Genetic variations affecting IL-10 production and the genotypes affecting IL-10 serum levels are associated with the occurrence of asthma and are attributed to the IL-10 -1082G/A promoter gene polymorphism. There was a strong correlation between cytokine levels, of disease development, and the genotypes of the AA and AG genes, indicating that IL-10 -1082A/G predisposition to asthma may be influenced by the gene promoter polymorphism. Asthma development and immunological markers (IL-10) are substantially correlated. One theory links allergic rhinitis to both the development of asthma and its risk. Inducing long-term immunological and clinical tolerance in patients was a good use of HDM immunotherapy. The current study's findings indicate a substantial difference between the asthma patients and the control group in terms of gene type and allele frequency of the IL-6 -174G/C polymorphism. The patients exhibited a higher prevalence of the G allele and the GG homozygous genotype than the control group. Therefore, it was shown that those with GG genotypes had a 2-fold increased probability of having asthma, indicating that patients were more prone to the condition.

From natural marine bacteria and fungi to culturable ones: What affects their bioactivity?

Endah Puspitasari, Kartika Senjarini , I Dewa Ayu Ratna Dewanti, Antje Labes

APJMBB 32(3): 56-66

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.07

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Marine bacteria and fungi have been known as sources of drug candidates. Novel natural products discovery derived from marine bacteria and fungi has been boosted in the last decades. This review discusses the challenges of culturing marine bacteria and fungi taken from their natural habitat into laboratory-scale culturable ones. Since, not all bacteria and fungi, when they are taken from their natural habitat and grown in a laboratory setting, are culturable. This review will also discuss possible solutions to overcome those challenges. Targeted bioactivity relies on the marine bacterial and fungal growth, especially due to the capability on producing the intended natural products. From this review, some important factors that must be taken into consideration to have optimal bacterial and fungal growth are genes and their variation in a species, media composition, and growth condition.

Carbon - ¹²C ion radiation-induced cell death and molecular mechanisms in malignant cells: Evidence from in vivo and in vitro

Mriganka Mandal

APJMBB 32(3): 67-87

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.08

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Conventional radiotherapy has been revolutionized for several decades. Recently, the delivery of high linear energy transfer (LET) radiation such as heavy carbon ion therapy has gained more attention. Among all low-energy therapies such as photon, X-ray, and gamma radiation therapy, ¹²C heavy carbon ion therapy is the best therapy for cancer treatment due to its high relative biological effectiveness (RBE). It has been studied that many tumors show resistance to chemotherapy and low energy transfer radiation. The unique nature of high LET shows unique Bragg peaks where very little radiation are applied on the tumor cells and better results can be achieved. It effectively kills the tumor cells without affecting the normal cells. There are some limitations to conventional radiotherapy. Radiotherapy with the low LET group may not work properly. Carbon ion radiation (CIR) induces malignant cell death in different modes, such as apoptosis, senescence, autophagy, immunogenic cell death and necrosis. Heavy carbon ion therapy enhances cell death by altering different molecular signalling pathways such as p53, AKT, MEK, caspase and STAT–3. Investigation of the major molecular cell death by carbon ion irradiation is very essential to understand the modalities of heavy carbon ion induced cell death. In this context, the investigation of different signaling cascades may open new horizons in cancer treatment. The therapeutic efficacy and application of heavy carbon ion beams along with some molecular inhibitors, radio sensitizers, and gene therapy would be helpful for future cancer treatment. Therefore, a more thorough investigation in this field is needed.  The current review article highlights the recent advances in cancer therapy research based on Carbon ¹²C Ion radiation-induced cell death.

Detection of fim H and UDP virulence-associated genes in patients with MDR Klebsiella pneumoniae isolates from Baghdad hospitals

Zahraa Redha Shamsee, Aida Hussain Ibrahim

APJMBB 32(3): 88-97

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.09

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Klebsiella pneumoniae is a gram-negative, encapsulated, and lactose-fermenting bacterium related to hospital-acquired infections that can cause various diseases. The current study aimed to detect the presence of the virulence-associated genes fim H and UDP among multiple drug-resistant K. pneumoniae isolates obtained from different sources using polymerase chain reaction (PCR) to determine their association with antibiotic resistance. Ninety K. pneumoniae isolates were obtained from 210 different clinical samples collected from various sources (urine, sputum, wound swabs, blood, and burns) from patients of both genders in Baghdad hospitals between March 2022 and July 2022. Bacterial identification was based on selective and differential media, followed by seven biochemical tests, the Vitek-2 system, and molecular identification tests were performed using 16S rRNA. The antibiotic susceptibility of isolates to 13 antibiotics was assessed by the disk diffusion method on Mueller–Hinton agar. The results showed that 100% of the isolates were resistant to vancomycin. Otherwise, colistin had the lowest resistance rate (25.55%). The virulence genes fim H and UDP (uge) were detected by multiplex polymerase chain reaction (PCR) in multidrug-resistant (MDR) isolates. The most prevalent gene was fim H (88%) followed by UDP (80%). The virulence genes were highly prevalent among MDR K. pneumoniae isolates. The resistance rate to colistin was 25.55%, making it the most effective antibiotic used in the current study. Vancomycin, on the other hand, had a high resistance value. Multidrug resistant isolates were found to be associated with the presence of virulence genes.

Effect of probiotics, prebiotics, synbiotics on survival, growth, and immune response of white shrimp (Litopenaeus vannamei) infected with Vibrio parahaemolyticus

Deshinta Arie Widyany, Slamet Budi Prayitno, Desrina

APJMBB 32(3): 98-112

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.10

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Vibrio parahaemolyticus is one of the causative agents of vibriosis which causes mortality and significant economic losses in shrimp aquaculture. Application of probiotics, prebiotics, synbiotics is an environmentally friendly strategy to control vibriosis and limit the use of antibiotics. This study aimed to evaluate the impact of probiotic, prebiotic and synbiotic diets on survival, growth and immune response of white shrimp. The experiments consisted of 5 treatments and 5 replications. (K-) Without the addition of probiotics, prebiotics, and synbiotics and no challenge test; (K+) Without the addition of probiotics, prebiotics, and synbiotics and challenge test; (P1) Addition of probiotics and challenge test; (P2) Addition of prebiotics and challenge test; (P3) Addition of synbiotics and challenge test. The parameters measured were SGR, FCR, THC, phagocytosis activity, lysozyme activity and SR. The results showed that SGR and FCR with probiotic (4.70±0.29%; 1.53±0.10) and synbiotic (4.60±0.15%; 1.64±0.14) diets were significantly different than K+ and no significant differences between them. The post-challenge results showed that the survival with probiotic (61.33±8.69%), prebiotic (42.67±7.60%) and synbiotic (57.33±10.11%) diets was significantly different than K+ (21.33±11.93%). No significant differences between probiotic and synbiotic diets on survival. Synbiotic diets showed the best immune response post-challenge test with THC (1.59±0.26x10⁶ cells/mL), phagocytosis activity (42.80±6.91%), and lysozyme activity (0.92±0.11 cm) and significantly different than K+. The study concluded that probiotic or synbiotic diets had the best effect on SGR, FCR and survival of white shrimp post V. parahaemolyticus infection. Meanwhile, synbiotic diets showed the best immune response post V. parahaemolyticus infection.

Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W

Siti Hajar Yusof, Adam Thean Chor Leow, Raja Noor Zaliha Raja Abd Rahman, Mohamad Syazwan Ngalimat, Si Jie Lim, and Suriana Sabri

APJMBB 32(3): 113-127

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.11

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Given its thermoalkaliphilic properties, T1 lipase holds significant potential for diverse industrial applications. However, traditional expression methods in Escherichia coli, specifically the plasmid-based system, present challenges of exerting metabolic burden on host cells and elevated costs due to antibiotic usage. This study addresses these issues by pioneering the expression of recombinant T1 lipase in a sucrose-utilizing E. coli strain W, using molasses as an economical carbon source. The gene cassette (KIKO plasmid), containing the T1 lipase gene regulated by tac and trc promoters, was integrated into the E. coli genome via the λ Red recombinase system. T1 lipase was optimally expressed in shake flasks at 16°C and a 3% molasses concentration in M9 medium with 0.8 mM IPTG as inducer, yielding 0.44 U/mL activity in the genome-based system compared to 0.94 U/mL in the plasmid-based system. This study not only underscores the potential of employing sucrose-utilizing E. coli strain for industrial recombinant protein production but also highlights the need for further optimization of genome-based expression systems. It offers an alternative to reduce costs and enhance sustainability in the stable production of industrially relevant enzymes like T1 lipase, without the need for antibiotic supplementation, and has broader implications for leveraging inexpensive carbon sources like molasses in biotechnological applications.

Surface functionalization of oil palm empty fruit bunch (OPEFB)-derived cellulose as a carboxyl platform for metal cations and dyes removal from aquatic media

Noerhidajat Sjahro, Robiah Yunus, Luqman Chuah Abdullah, Suraya Abdul Rashid, Ahmad Jaril Asis, Dina Kania, Alsultan Karim

APJMBB 32(3): 128-146

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.12

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The abundant oil palm empty fruit bunch (OPEFB) as by-product of palm oil milling processes exhibits a potential as an alternative cellulose feedstock for bio-adsorbent. This study aimed to produce a highly carboxylated bio-adsorbent for direct industrial application from OPEFB-based cellulose via mercerization and followed by esterification with succinic anhydride (SA) to enhance its adsorptive capability towards hazardous heavy metal and dyes ions. The modification using SA provides the carbon backbone platform for carboxyl group attachment for the contaminants. The results showed that the carboxylated cellulose had a high carboxyl content (4.39 mmol/g). Carboxylated cellulose had a higher binding capacity for adsorbates, with removal rates of 94.7%, 97.85%, 40.9%, and 90.15% for dye, Pb2+, Cu2+, and Cd2+ cations, respectively, at pH 6, 4 hours reaction time, and at room temperature. In comparison, unmodified cellulose removed only 47%, 23.1%, 2.9%, and 7.5% for dye, Pb2+, Cu2+, and Cd2+ cations, respectively. The adsorption kinetics study revealed that the adsorption process followed the pseudo-second-order model. The adsorption isotherm of these two metal cations follows the model of Langmuir very well, while Cu2+ follows the Freundlich model. Our method produces bio-adsorbents with high carboxyl content and adsorption rate in a short reaction time using OPEFB as a green precursor material that is easily scalable for industrial use.

Evaluation of the antibacterial activity of pineapple (Ananas comosus) (L.) Merr. industrial waste against common fish and shellfish pathogens

Honielyn Taka, Fiona L. Pedroso, Casiano H. Choresca, Jr., Christopher Marlowe A. Caipang, Fernand F. Fagutao

APJMBB 32(3): 147-156

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.13

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Disposal of industrial pineapple (Ananas comosus) wastes is a pressing environmental issue due to pollution risks when accumulated in large quantities. These wastes are susceptible to microbial spoilage, posing serious environmental and health concerns. Therefore, exploring their conversion into valuable products is crucial for effective waste management. In this study, we assessed the antibiotic properties of pineapple processing wastes against prevalent fish pathogens in aquaculture. The wastes underwent various drying methods: sun-drying (SD), oven-drying (OD), and mechanical dehydration (DH). Ethanol extraction was used to isolate bioactive compounds, which were then tested for antibacterial activity at a concentration of 1000 mg/ml using the Agar Well Diffusion technique and Zone of Inhibition (ZOI) assay. Minimum Inhibitory Concentration (MIC) values were determined across six concentrations: 31.25 mg/ml, 62.50 mg/ml, 125 mg/ml, 250 mg/ml, 500 mg/ml, and 1000 mg/ml. Results indicated that all extracts from different drying processes effectively inhibited all tested aquaculture pathogens. DH extracts showed the highest antibacterial activity against Vibrio harveyi and Vibrio parahaemolyticus, with ZOI of 24.67 + 1.25 mm and 21.67 + 0.47 mm, respectively, and a consistent MIC of 250 mg/ml across all pathogens. SD extracts displayed a MIC of 125 mg/ml against Streptococcus agalactiae, while OD extracts showed a MIC of 1000 mg/ml against Edwardsiella tarda, Aeromonas veronii, and S. agalactiae, and 250 mg/ml against both Vibrio species. Comparative analysis with oxytetracycline did not reveal significant differences. These findings suggest that pineapple waste extracts have potential as natural antibacterial agents against common aquaculture pathogens, offering an eco-friendly alternative to commercial antibiotics.

Effects of silver nanoparticle on Morinda officinalis How

Mai Van Hung, Nguyen Hoang Vu, Phuong Thi Mai Trang, Tran Thi Minh

APJMBB 32(3): 157-165

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.14

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Nanosilver is a form of silver metal particles, extremely small nano-sized particles, about 1 to 100 nm. Nanosilver solution can be used to treat the soil before sowing or planting trees to control pathogenic microbes and improve plant growth and development. Nevertheless, the study on the effects of silver nanoparticles remains limited. Therefore, the aim of the current study was to examine the effects of nanosilver concentration on the growth and development of Morinda officinalis How grown in Ba Che district, Quang Ninh province. The silver nanoparticle solution (2 to 10 ppm) mixed with the extract of sour star fruit was sprayed on the seedlings of M. officinalis How and their growth and development were measured. The results showed that the use of nano-silver solution affected the growth and development of Morinda officinalis How. The study successfully prepared a 500 ppm nanosilver solution by a chemical reduction method with reducing agent extracted from the sour star fruit. It has the dual effect of both promoting growth and development as well as producing the best tuber quality for Morinda officinalis How grown in Ba Che, Quang Ninh at 6 ppm.

The upregulation of protein disulfide isomerase (PDI) and its potential usage in drug-targeted therapy for breast cancer

Lay-Harn Gam, Chu-Ai Lim, Saad Mardi Muhammad, Boon-Yin Khoo, Manjit Singh

APJMBB 32(3): 166-181

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.15

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Breast cancer is the leading cause of cancer death among women worldwide. As of the end of 2020, there were 2.3 million women diagnosed with breast cancer and 685 000 deaths globally.  Chemotherapy is a common treatment for breast cancer although it was known to be associated with many side effects.  It is believed that such treatment can be improved by drug targeted therapy. Recently we have carried out a preliminary study on proteomics analysis of 25 pair of surgically removed breast cancerous tissues and normal tissues from patients.  Differentially proteins expression between the types of tissues was done by 2D-gel electrophoresis separation followed by protein profiles mapping. The identity of the targeted protein spots was analysed by LC/MS/MS and protein database search. The data was then confirmed by Western blots. Subsequently, immunocytostaining experiments were carried out to determine the cellular location of the targeted proteins. A few proteins were found significantly (p <0.05) upregulated > 2 folds in breast cancerous tissues compared to breast normal tissues.  Two of the up-regulated proteins, namely HSP60 and PDI were upregulated in stage 2, stage 3, T2, T3, N2, and N3 breast cancers. The immunocytostaining revealed the extracellular location of these proteins, while the strong immunoreactivity of PDI with its anti-PDI antibody marked it as a usefulness target for breast cancer therapy.

Indigenous copper and dye resistant bacteria: Enterobacter cloacae Suk1 and Serratia nematodiphila Suk13 isolated from Sukolilo River, Surabaya Province, Indonesia

Wahyu Irawati , Reinhard Pinontoan, Triwibowo Yuwono, Valentine Lindarto, Candra Yulius Tahya

APJMBB 32(3): 182-190

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.16

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Bioremediation using indigenous copper-resistant bacteria has been successfully used in reducing copper concentrations. However, little information is available concerning the resistance of bacteria to copper and dyes. This study, therefore, was aimed at 1) isolating and characterizing multi-resistant bacteria, 2) measuring the copper biosorption and accumulation ability, and 3) measuring the growth and decolorization ability of various dyes. Dye-multi-resistant bacteria were isolated from Sukolilo River, Indonesia. Copper resistance was determined by measuring the Minimum Inhibitory Concentration (MIC). The biosorption and accumulation abilities were measured using an atomic absorption spectrophotometer. The twelve dyes used in the test were methylene blue, malachite green, congo red, mordant orange, reactive black, direct yellow, basic fuchsine, reactive orange, dispersion orange, remazol red, wantex yellow, and wantex red. The decolorization activity was analyzed by spectrophotometry at a wavelength of 300-900 nm. The results showed that nine isolates of copper-resistant bacteria demonstrated MIC of 3-9 mM CuSO₄. Enterobacter cloacae Suk1 and Serratia nematodiphila Suk13 have been demonstrated to possess multi-resistance to CuSO₄, and the twelve dyes, except Enterobacter cloacae Suk1 which did not grow on malachite green and basic fuchsine. Enterobacter cloacae Suk1 was able to decolorize 89.42% of methylene blue and 83.61% of congo red in a medium supplemented with 500 ppm of each dye. Enterobacter cloacae Suk1 and Serratia nematodiphila Suk13 also accumulated copper of up to 2.61 mg and 2.48 mg/g dry weight of cell, respectively, and removed copper of up to 94.64% and 90.52% in a medium containing 5 mM CuSO₄, respectively. 

Development of selectable markers for mitochondrial transformation in yeast

Mei San Ho, Wai Keat Toh, Shu Ting Chang, Kiao Huio Yap, Pek Chin Loh, Parameswari Namasivayam, Hann Ling Wong

APJMBB 32(3): 191-205

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.17

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Mitochondria, present in most eukaryotic organisms, are crucial for energy production and essential for cellular functions. Sequencing of the complete mitochondrial genome of Saccharomyces cerevisiae in 1998 has paved the way for mtDNA gene editing, enabling the study of mitochondrial function and potential gene therapies for mitochondrial diseases. Effective selectable markers are crucial for addressing heteroplasmic mtDNA issues after mitochondrial transformation. Antibiotic resistance (Ab^R) marker genes aadA1, cat, and hph confer resistance to streptomycin, chloramphenicol, and hygromycin B, respectively. This study aimed to explore the feasibility of employing these Ab^R markers for selecting transformed yeast cells. Additionally, the usefulness of these Ab^R genes as selectable markers for yeast mitochondrial transformation was assessed by fusing a mitochondrial targeting signal (MTS) to the N-terminus of these genes using overlapping PCR.  The minimal inhibitory concentration (MIC) of yeast transformants expressing various Ab^R genes, with or without MTS fusion, was determined using the agar dilution method. Yeast transformants expressing aadA1, cat, and hph, with or without MTS fusion, displayed resistance to streptomycin (>10 mg/mL), chloramphenicol (up to 6 mg/mL), and hygromycin B (up to 4 mg/mL), respectively. MICs were similar between Ab^R and MTS-tagged Ab^R yeast transformants. To assess mitochondrial targeting, GFP was fused to the C-terminus of cat and MTS-cat gene constructs. Fluorescence microscopy confirmed MTS-tagged CAT-GFP localization to yeast mitochondria, while CAT-GFP showed cytoplasmic localization. The fluorescence microscopy results were confirmed by Western blotting. This study demonstrated that yeast transformants expressing aadA1 exhibit a significant level of streptomycin resistance (>10 mg/mL), suggesting that aadA1-mediated streptomycin resistance has the potential to serve as a selectable marker for mitochondrial transformation in yeast.

Production of plantain powder using enzyme-liquefied feed through spray drying operation

Lee Sin Chang, Khu Lee Liew, Liew Phing Pui

APJMBB 32(3): 206-214

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.18

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Plantain is a tropical fruit that is widely consumed due to its exotic flavor and nutritional value. However, the fresh plantain is very perishable, in which it has a short shelf life i.e., 10 days when stored at room temperature. Hence, the conversion of the fresh fruit into a value-added product such as powder can prevent post-harvest loss. To do this, the plantain fruit was undergoing enzymatic treatment to produce a low-viscosity feed before the production of powder using the spray drying method. The enzymatic treatment with Celluclast ® at 2.0% (v/v) for 2 hours at 40°C produced feed with the lowest viscosity level (101.27cp) with the highest total soluble solid value (32.30°Brix). Then, 30% maltodextrin was added as a carrier agent for spray drying at different inlet temperatures of 160–200°C. From the results, the inlet temperature at 180°C was the optimized temperature with the yield of 26%. The properties of produced plantain powder had a water activity at 0.17aw, moisture content at 4.4%, water solubility index at 89.83%, water absorption index at 45.41%, color measurement 83.86, 1.05 and 11.16 for L*, a* and b*, respectively, hygroscopicity at 0.07%, proximate composition of protein at 0.029%, fat at 4.391%, crude fiber at 0.025% and ash at 9.610%. In a conclusion, spray-dried plantain powder is able to minimize the disposal of waste from plantain plantations and broaden the application of plantain powder to be applied into various innovative food products.

Cissus hastata (Semperai) adverse anti-inflammatory response via cytokines expression with moderate cytotoxicity in vitro and in vivo towards haemorrhoid

Sharaniza Ab-Rahim, Wee Ai Sze, Wan Nor I’zzah WMZ, Jesmine Khan, Nora Asyikin Ramli, Nur Sufiah Zulkifli, Mudiana Muhamad

APJMBB 32(3): 215-223

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.19

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Cissus hastata (CH) or Semperai, is widely utilised by locals in Malaysia as antitussive, expectorant and flatulence reliever. However, scientific evidence on the pharmaco-active property of CH remains elusive, although previous studies reported the gastro-protective action of another cissus species, Cissus quadrangularis among haemorrhoids patients. Hence this study aims to investigate the anti-inflammatory action of CH towards cytokine expression. The in vitro analysis involved Caco-2 cells via cytotoxicity assay of CH crude extract and ELISA of TNF-α and IL-6 expression levels. The in vivo analysis involved haemorrhoid-induced animal model (Wistar rats). Result of Cell WST-8 assay for CH showed high range of IC₅₀ values (>30 μg/mL), indicating moderate to high cytotoxicity. The ELISA revealed increased expression of both TNF-α and IL-6 in CH post-treated Caco-2 cells, suggestive of non-inhibitory response against the cytokines. Nevertheless, better appearance of tissue morphology was observed in the haemorrhoid-Wistar rat group that received 200 mg/kg of CH, indicated by the absence of necrotic cells and presence of inflammatory cells. In conclusion, the in vitro results provide preliminary evidence on the intricate biological action of CH at the cellular level while the in vivo result clearly disclosed the beneficial outcome of CH as treatment for haemorrhoid.

Harnessing of low cost agro-industrial waste substrates for the optimum production of citric acid: A review

Geeta Rawat , Vidhi Kothari, Neha Giri, Parul Sharma

APJMBB 32(3): 224-235

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.20

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Citric acid, an essential organic acid, holds significant importance across various industries due to its multifaceted applications in sectors such as food, detergents, pharmaceuticals, and cosmetic. A broad range of microbes have potential to produces citric acid in optimum concentrations, most importantly includes Aspergillus niger, Aspergillus flavus, Candida tropicalis, saccharomyces cerevisiae, and Yarrowia lipolytica. This review explores the biosynthesis of citric acid through innovative utilization of low-cost waste substrates and presenting an eco-friendly approach towards its synthesis. Subsequently, the classical and conventional production processes of citric acid are usually energy-intensive procedures, time consuming and utilizing expensive raw materials. In contrast, the utilization of low-cost waste substrates emerges as a sustainable alternative, promising reduced production costs and harmless for environmental. Various waste substrates, ranging from agricultural residues to industrial by-products, are evaluated for their potential in citric acid production. Since, their abundance, affordability and untreated or unutilized high nutrient value it gains a high research attention. This review emphasizes on a paradigm shift towards utilizing and exploring low-cost waste substrates and from where it can be collected.

Functionality of XVE-inducible system and toxicity assessment of the bacterial PezT toxin in microalga, Messastrum gracile SE-MC4

Sik Ze Gan, Chew Chieng Yeo, Thye San Cha

APJMBB 32(3): 236-247

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.21

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This study investigated the effects of the heterologous expression of the pezT toxin gene from the bacterium Streptococcus pneumoniae in the green microalga, Messastrum gracile SE-MC4. The microalgae was co-transformed with the pMDC150_35S activator vector (containing the XVE expression cassette) and the pMDC221_pezTGFP responder vector (containing pezT-GFP fusion genes) using Agrobacterium-mediated transformation. The stable transgenic line, M. gracile SE-MC4 (pezT-GFP) was confirmed through antibiotic selection and PCR validation. Upon induction with 17-β-estradiol, GFP signals were observed in the transgenic M. gracile SE-MC4 (pezT-GFP) cells, indicating successful transformation and expression of the transgene cassettes. Furthermore, the functionality and action of the PezT toxin was demonstrated with a significant reduction of 52% in cell viability of transgenic cells, compared to wild-type cells. This finding suggests the potential use of bacterial toxins as a novel approach for harvesting microalgal cellular contents for various microalgae-based biotechnological applications. It also can be a containment system to prevent the accidental release of transgenic microalgae and used for transgenic microalgae selection, as an alternative to antibiotic selection currently widely used.

Impact of ZnO nanoparticles on MagA gene expression in Klebsiella pneumoniae isolated from diabetic foot ulcers

Mustafa Muhammed Jadooa, Haider Turky Mousa Al-Mousawi, Nadhim Mushtaq Hashim Al-bderee

APJMBB 32(3): 248-256

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.22

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Diabetes mellitus (DM) is an accumulation of glucose in the blood. Diabetic mellitus is divided into gestational diabetes, type 1 and type 2. Complications of diabetes mellitus include diabetic foot ulcers. The most important complication of diabetic foot ulcers is bacterial infection, which leads to gangrene when various types of antibiotics fail to prevent bacterial infections. To explore the effect of synthetic zinc oxide nanoparticles (ZnONPs) on the expression of the MagA gene in Klebsiella pneumoniae isolated from diabetic foot ulcer cases. ZnONPs were biosynthesized by Aspergillus niger and characterized using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), X-ray diffraction (XRD), and UV-visible spectroscopy. The inhibitory effect of ZnONPs on K. pneumoniae isolates was examined by determining the minimum inhibitory concentration (MIC) of the ZnONPs. Besides, the values of the MagA gene before and after exposure to ZnONPs would be examined by RT-qPCR, with 16SrRNA gene as the reference gene. According to SEM and XRD results, the ZnONPs are in the form of nanocrystals and have small spherical crystals with an average size of 13.5 nm. The highest optical density of the synthesized nanoparticles was obtained at 390 nm. The biosynthesized ZnONPs exhibit a strong inhibition zone against K. pneumoniae corresponding to 125 μg/mL. The fold of gene expression ranged from 0.042 to 0.118. The research reveals that ZnONPs exhibit strong antibacterial potential and affect gene expression in K. pneumoniae strains. The results of the present study indicate that ZnONPs can indeed be used in the treatment of infectious bacteria.

Optimization of non-effervescent riboflavin gastroretentive floating tablets using mixture design

Umme Tabassum Arobi Katha, Muhammad Sofwan Sapiyan, Fatini Nasuha Ahmad Puad, Nik Sofea Aliya Nik Aris, Mohd Akmal bin Azhar

APJMBB 32(3): 257-266

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.23

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Gastroretentive Floating Drug Delivery Systems (GRFDDS) are long-acting oral dosage forms that float on gastric juice and remain in the stomach for an elongated period gradually delivering drug substances to the upper part of the gastrointestinal system. This study aims to develop and enhance the bioavailability and stomach retention of non-effervescent riboflavin floating tablets by using a variety of polymers. In this investigation, both pre-compression evaluation and post-compression of all the tablet materials were performed according to USP specifications. In vitro, buoyancy analyses were carried out to achieve minimum floating lag time and maximum floating duration. The tablet employed direct compression methods using HPMC K17, Carbopol 940p, and polypropylene foam powder. In vitro, buoyancy studies were performed to achieve minimum floating lag time and maximum floating duration. Tablets were evaluated for physicochemical properties according to USP specifications. An optimized tablet with a floating lag time of 0.77 minutes and a floating time of 48.74 minutes was developed using Design of Experiments (DoE). The results indicated that the optimized formulation, designated as Y, performed the best. It consists of 0.45% polypropylene foam powder, 0.15% HPMC K17, and 0% Carbopol 940p. The developed non-effervescent riboflavin floating tablets have the potential to improve the bioavailability and therapeutic efficacy of riboflavin by enhancing its gastric residence time.

Screening of microbial population in Sabah tea kombucha pellicle for its potential as prebiotic and probiotic supplement

Nurul Farhana Nasir, Nurul Elyani Mohamad, Chee Wei Yew, Suryani Saallah, Noorjahan Banu Alitheen

APJMBB 32(3): 267-274

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.24

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This research aimed to determine and analyze the microbial population in kombucha pellicles derived from Sabah black tea, specifically focusing on bacteria and yeast, to gain insights into their abundance, diversity, and potential as prebiotic and probiotic supplements. Despite the growing interest in kombucha in Malaysia, the specific microbial composition of the pellicle from locally sourced Sabah black teas remains underexplored. Understanding this composition could reveal its potential as a sustainable source of health-promoting microbes. It is hypothesized that Sabah tea kombucha pellicle harbours a beneficial microbial population that can be utilized as a low-cost prebiotic and probiotic supplements. The kombucha pellicle was prepared using 10 g of Sabah black tea, 1 L of sterile water containing 10% sugar (w/v), and a 10% kombucha symbiotic culture of bacteria and yeast (SCOBY). The process included boiling black tea with sugar, adding SCOBY culture, and allowing fermentation for 30 days to obtain cellulosic pellicles. After fermentation, the pellicle was separated, homogenized, and stored for further use. Then, kombucha pellicle genomic DNA was extracted and subjected to 16S and ITS metagenomic analysis to identify the bacteria and fungi population. The 16S and ITS metagenomic results showed that Sabah tea kombucha pellicle contains a potentially beneficial microbial population, mainly Komagataeibacter, Zygosaccharomyces and Starmerella, that may serve as a sustainable probiotic. This current study provides promising evidence for using Sabah tea kombucha pellicle as a low-cost prebiotic and probiotic supplement. This will indirectly help advertise and commercialize Sabah tea as one of the local products in Sabah.

A novel method for the degradation of human blood clot by immobilised bromelain using multi-walled carbon nanotube and polyphenol oxidase

Ali J. R. Al-Sa'ady, Zainab W. Abdulameer, Sumaya Saady, Mohammed J. Al-Obaidi , Mohanad S. Al-Fayyadh, Haider A. Hassan, Kefah H. Ismael, Ahmad Y. Hanoon, Iftikhar A. Alqaissy

APJMBB 32(3): 275-288

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.3.25

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Pathological blood clot in blood vessels, which often leads to cardiovascular diseases, are one of the most common causes of death in humans. Therefore, enzymatic therapy to degrade blood clots is vital. To achieve this goal, bromelain was immobilized and used for the biodegradation of blood clots. Bromelain was extracted from the pineapple fruit pulp (Ananas comosus) and purified by ion exchange chromatography after precipitation with ammonium sulphate (0-80 %), resulting in a yield of 70%, purification fold of 1.42, and a specific activity of 1175 U/mg. Bromelain was covalently immobilized on functionalized multi-walled carbon nanotubes (MWCNT), with an enzyme loading of 71.35%. The results of the characterization of free and immobilized bromelain demonstrated that the optimum pH for free and immobilized bromelain activity was 7.0, while the pH range of stability was from 5.0 to 8.5 and 4.0 to 9.0, respectively. The optimum temperature for free and immobilized bromelain activity was 45ºC, whereas the stability was 15 to 50°C and 15 to 55°C, respectively. The immobilized bromelain activity was decreased after the fifth reuse, and the storage period of the free and immobilized bromelain was decreased after 6 and 123 days, respectively. Casein was the best substrate-free bromelain, and fibrin was the best substrate for immobilized bromelain. The results of the purification of polyphenol oxidases (PPO) from potatoes by ion exchange chromatography gave a yield of about 54 %, a purification fold of 1.27, and a specific activity of 2804 U/mg. The current study showed that the immobilized bromelain can significantly biodegrade human blood clots in vitro, while the PPO enzyme has no significant effect on blood clots.

Volume 32(4); 2024

Molecular identification and phylogenetic analysis of coral and megabenthic species from Gede Island, Central Java, Indonesia

Yuyun Khoirun Nisak, Diah Ayuningrum , Pujiono Wahyu Purnomo

APJMBB 32(4): 1-9

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.01

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Coral reef ecosystems are home to many organisms, including bivalves, gastropods, crinoids, and demersal fish, making biodiversity assessment crucial for effective conservation and management. This study aims to: i) apply DNA barcoding to identify coral and megabenthic species in coral reef ecosystems, and ii) construct phylogenetic tree to trace their evolutionary relationships. Using quantitative descriptive methods and purposive sampling, specimens were collected in July 2023. The study yielded one coral (R2), one hydrozoan (R3), and one gastropod (GP-1) sample were successfully barcoded. DNA barcoding through Cytochrome Oxydase I (COI) gene amplification revealed that sample R2 had a 98.70% similarity with Acropora hyacinthus, sample R3 had a 98.92% similarity with Zanclea galli, and sample GP-1 had a 91.19% similarity with Erronea errones. Phylogenetic analysis positioned R2 in the same clade as Acropora hyacinthus and related genera such as Montipora, Astreopora, and Alveopora, within the Acroporidae family. R3 clustered with Zanclea galli in the hydrozoan group, alongside Z. sesillis, Z. implexa, and Z. sango. GP-1 grouped with Erronea errones in the gastropod genus Erronea, showing relationships with other genera such as Blasicrura, Cribrarula, Bistolida, and Eclogavena within the subfamily Erroneinae. Species inventories like this are essential for the conservation and management of coral reefs, which are key habitats for fish and other marine life.

Determination of some mycotoxins from poultry feed in Baghdad city

Aida Hussan Ibrahim, Dunya Dhafer Taher, Oday Sattar Abbas, Dalia Abdul-Kareem Abdul-Shaheed

APJMBB 32(4): 10-15

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.02

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The aims of this study were to determine some mycotoxins and to isolate the fungi from chicken feed samples from broiler and layer farms in Baghdad city. The isolation rate (percentage) of fungi from poultry feed samples was Aspergillus niger 17.7%, Aspergillus flavus 14.7%, Aspergillus fumigatus 12.4%, Aspergillus ochraceus 10.7%, Penicillium spp. 9.3%, Fusarium spp. 9.3%, Rhizopus spp. 8.4%, Mucar spp. 8.4%, Absidia spp. 7.6% and Chrysonilia sitophila 1.5%, while the most common yeast genera were Candida spp. 44.8%, Candida albicans 20.7%, Cryptococcus neoformans 17.3%, Geotricum candidum 12% and Trichosorom spp. 5.2%. The mycotoxins ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone (ZON) were detected, and the percentages of these mycotoxins were 43.1%, 27.2%, 21.1% and 8.6%, respectively. This study indicates that the presence of mold that produce mycotoxins in poultry feed increases the risk of mycotoxins in feed, meat, eggs, etc. of animals and therefore humans, and we need to raise awareness of the Ministry of Agriculture and Health to prevent these measures to reduce the mycotoxin levels in poultry products. 

In silico sequential mutagenesis of the Carbohydrate Binding Module family 32 (CBM32) enhances ligand binding affinity

Nazmi Harith-Fadzilah, Rosli Md Illias, Nadiawati Alias

APJMBB 32(4): 16-28

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.03

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Alginate lyase is a promising target for genetic modification for its degrading biofilm, contributing to bacterial proliferation and antimicrobial resistance. Apart from the main enzyme, the carbohydrate binding module (CBM) component can also be modified to enhance alginate lyase’s activity. This study aimed to perform sequential in silico mutagenesis, molecular docking of selected amino acid residues of Vibrio splendidus CBM32 and performed molecular dynamics (MD) simulations of the mutated structure to validate its ligand-binding efficacy. Seven residues were selected for mutagenesis based on the predicted bonds that formed between the CBM32 and the glucuronic acid ligand (LGU9). Four of seven sequential residue substitutions increased the ligand binding affinity cumulatively from -5.4 Kcal/mol to -6.9 Kcal/mol. The mutated CBM32 had similar MolProbity scores to the original V. splendidus CBM32 structure. From the post-MD simulation analysis, the mutated CBM32 had higher structural stability in a solvent system, a greater number of hydrogen bonds formed with ligand but a lower solvent-accessible surface area than the original structure. The sequential mutagenesis process significantly increased the ligand binding affinity of CBM32 while incurring a minimal change in the overall CBM32 structure. The information on these substituted residues would be beneficial for designing subsequent in vitro mutagenesis and enzymatic assays.

Environmental enrichment preconditioning prevents chronic social defeat stress-induced memory impairment and hippocampal neurodegeneration

Nik Nasihah Nik Ramli , Nur Adzreen Shamsul Adzmi, Siti Nur Atiqah Zulaikah Nasarudin, Mohamad Hisham Hashim, Maisarah Abdul Mutalib, Muhammad Najib Mohamad Alwi, Aswir Abd Rashed, Rajesh Ramasamy

APJMBB 32(4): 29-38

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.04

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The World Health Organization anticipated major depressive disorder to become the primary contributor to the global burden of disease by 2023. Chronic psychological stress directly impacts synaptic plasticity and the formation of stress-associated memories, contributing to a predisposition to depression. Conversely, research suggests that environmental enrichment, promoting physical activity, social interaction, and sensory stimulation, positively influences neurocognitive functions. However, there is a paucity of studies examining the effects of environmental enrichment on learning and memory in chronic social defeat stress (CSDS) models. Male C57BL/6N mice were (n=5) housed in enriched environments for 30 days before undergoing daily social defeat stress by ICR strain aggressor mice over 10 days. Depressive-like behavior was assessed using the forced swim test (FST) and tail suspension test (TST). Memory and learning functions were evaluated through the Y-maze and novel object recognition (NOR) tests. Brain tissues were subjected to Hematoxylin and Eosin staining to examine histological alterations in the hippocampal region. Environmental enrichment significantly alleviated depressive-like behavior in mice subjected to CSDS. Dysfunctions in spatial working memory and recognition memory as well as hippocampal degeneration induced by CSDS were notably improved in mice exposed to environmental enrichment as compared to those without exposure. This study highlights the significance of environmental enrichment preconditioning in ameliorating memory and learning impairments induced by CSDS.

Assessing the accuracy of cystatin C for estimating glomerular filtration rate in patients with kidney disease

Ayad Abdull-Khaleq Ismael, Abdulla Adill Raoof, Yildiz Hasan Tahseen

APJMBB 32(4): 39-45

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.05

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The diagnosis, staging, and prognosis of chronic kidney disease (CKD) depend on the measurement of the glomerular filtration rate (GFR). A low GFR is associated with an increased risk of cardiovascular disease and early mortality. Current guidelines for CKD recommend using cystatin C as a supplementary test to ensure accurate estimation of eGFR. In the present study, we aimed to compare the efficacy of cystatin C-based eGFR with serum creatinine-based eGFR before and after contrast-enhanced intravenous urography (IVU). The present study is a retrospective observational study conducted in 51 patients aged 10 to 70 years. Of these patients, 21 were female and 30 were male, all of whom had kidney diseases and attended the hospital from January 2021 to January 2022. In this study, we evaluated the serum levels of urea, creatinine, and cystatin C before and after administering iodine solution to each participant. The study found no significant changes in blood urea and serum creatinine levels before and after administering the iodine solution. However, there was a significant variation in the levels of cystatin C. The levels of eGFR-dependent creatinine did not differ significantly, but there were significant changes in eGFR-dependent cystatin levels. Our findings suggest that cystatin C is a more accurate method of monitoring kidney function compared to creatinine levels. Additionally, cystatin C appears to be a more reliable marker of renal failure than serum creatinine and urea. 

Cytogenetic profile of leukemia cases in northern region of Malaysia - A single centre retrospective study

Nur Haida Natasha Shamsuddin, Abdul Rahman Azhari, Muhamad Amir Azizan, Hanis Nabilah Mohd Nazman, Zainul Abeden, Fadly Ahid, Narazah Mohd Yusoff & Asmida Isa

APJMBB 32(4): 46-54

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.06

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Leukemia is a heterogeneous disease in terms of cytogenetics, with four primary subtypes: acute lymphoid leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), and chronic lymphoid leukemia (CLL). As cytogenetic heterogeneity increases, the disease prognosis worsens, highlighting the significance of cytogenetic profile in disease diagnosis. In this study, we conducted cytogenetic profiling of 105 leukemia cases referred to the Clinical Diagnostic Laboratory (CDL) at the Advanced Medical and Dental Institute (AMDI) in northern Malaysia between 2006 and 2021. Of these cases, 50.47% were ALL, 37.14% were AML, and 12.38% were CML. Most of the patients, approximately 57.15%, were cytogenetically normal, while the rest, 42.85%, were cytogenetically abnormal. Overall, the most common cytogenetically abnormal karyotypes detected in patients were chromosomal translocation (20.95%), followed by complex karyotypes (13.33%), and chromosomal addition (4.76%). The majority of ALL patients were under 14 years old, whereas most AML and CML patients were older than 14. The correlation between the ages and the karyotype abnormalities in ALL, AML, and CML showed a negative moderate correlation (r=-0.501, p=0.312). In conclusion, cytogenetic profiling provides valuable insights into the disease's underlying mechanism, which may help strategize the treatment of leukemia patients.

Evaluation of the cytotoxic activity of essential oils from Malaysian herbal plants against A375 and A431 skin cancer cell lines

Bassam Mohamed Jawahar, Raha Ahmad Raus, Munira Shahbuddin, Mohd Rushdi Abu Bakar, Nur Najieha Binti Mohd Pauzi, Dayang Fredalina Basri, Irna Elina Ridzwan

APJMBB 32(4): 55-62

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.07

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Skin cancer incidences is rising due to prolonged harmful exposure to UV rays. Current therapies can adversely affect healthy cells. Conventional chemotherapy administered orally and intravenously poses risks of off targets, leading to severe and debilitating side effects. This study explored the anticancer potential of essential oils (EOs) extracted by hydrodistillation from seven local plant leaves (Piper betle, Persicaria minor, Cosmos caudatus, Citrus aurantifolia, Citrus hystrix, Citrus microcarpa, and Piper sarmentosum) on melanoma (A375), non-melanoma (A431), and fibroblasts skin cells (HFF1). Cytotoxicity assessment using MTT assay showed that all EOs tested had inhibitory effects of all EOs towards both cancer cell lines. Out of all seven plant EOs tested, P. sarmentosum EO displayed the most potent cytotoxicity at 80 ± 0.27 μg/mL with the extraction yield of 0.63%. Morphological changes in cellular size were observed after 4 h of EO treatment, leading to cell death coherently with declination of viability. This research signifies Malaysian plant’s role for topical application, as demonstrated by viability and morphology study.

Random amplified polymorphic DNA-based polymerase chain reaction is an effective tool to examine the genotoxic effects of some food colors

Doaa Adil Qasim, Inam Jasim Lafta

APJMBB 32(4): 63-73

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.08

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A large number of natural or synthetic dyes have been removed from both national and international lists of permitted food colors because of their mutagenic or carcinogenic activity. Therefore, this study aimed to use the Random Amplified Polymorphic DNA-Based Polymerase Chain Reaction (RAPD-PCR) assay as a feasible method to evaluate the ability of some food colors as genotoxin-induced DNA damage and mutations. Lactiplantibacillus plantarum was used as a bioindicator to determine the genotoxic effects by RAPD-PCR using M13 primer after treatment with some synthetic dyes currently used as food color additives, including Sunset Yellow, Carmoisine, and Tartrazine. Besides qualitative analysis, the bioinformatic GelJ software was used for cluster analysis to compare DNA fingerprints before and after treatment. The bacteria treated with the food colors showed the presence of polymorphism represented by DNA changes in the RAPD patterns, including variation in bands intensity, disappearance of normal bands, and appearance of new bands compared with the non-treated control. The GelJ program confirmed the presence of genetic variations between the bacteria treated with different concentrations of the food dyes and the bacteria without treatment. The RAPD approach can be applied for the detection of DNA damage and mutations induced by genotoxic compounds. Furthermore, L. plantarum and M13 are suitable as in vitro screening tools for detecting of potential genotoxicity of numerous compounds.

Epigenetic regulation of bovine embryo development: An update

Varsini, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu, Manoj Kumar Jena

APJMBB 32(4): 74-87

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.09

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Epigenetics is the study of heritable modifications in gene expression brought on by environmental influences including chemicals, stress, and diet that do not modify the DNA sequence. Traits including growth, reproduction, and disease resistance in farm animals are significantly influenced by epigenetic processes such as DNA methylation, histone modification, and non-coding RNA control. These mechanisms impact development rates, fat deposition, and milk production, according to studies. Long-term alterations are induced by environmental stresses including heat and malnutrition. Gaining the knowledge of epigenetic regulation can improve the efficiency and sustainability of livestock production by allowing producers to choose animals with desirable features without the modification of genes. Developing ethical and sustainable livestock management techniques that satisfy global demands while preserving the productivity and health of animals will require further research. This review article discusses about the effect of epigenetic regulation on well-being and productivity attribute of livestock.

Advances in employing omics and bioinformatics to reveal the potential of halophilic bacteria for biodegrading proteinaceous high-salt food waste

Ivan Kai Jie Lim, Chun Shiong Chong, Esther Zhe Hui Chong, Ming Quan Lam, Kah Yaw Ee

APJMBB 32(4): 88-96

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.10

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The waste biomass generated by the food processing industry poses significant concerns for environmental health. This mini review sheds light on the emerging challenge of proteinaceous high-salt food waste (PHFW) due to its high salinity. It aims to explore the potential of halophilic bacteria and their robust enzymes in addressing PHFW through advance approaches, including omics and bioinformatics. The integration of multi-omics and bioinformatics approaches could reveal the uncharted potential within the genomes of these halophilic bacteria, particularly their proteolytic capabilities. Furthermore, employing salt-tolerant proteases offers promising biotechnological applications for valorizing PHFW into valuable resources, such as bioactive peptides. The integration of advanced technologies not only elucidates the versatility of the halophilic proteases but also facilitates the development of functional food products from the subsequent hydrolyzed peptides. Though, transitioning these findings from laboratory settings to large-scale industrial applications presents various formidable challenges. In essence, the advancement in unlocking the potential of halophilic bacteria for biodegrading PHFW is emphasized as a potential bioeconomic solution.

Antibacterial activity of endophytic fungi Caulerpa racemosa against Vibrio parahaemolyticus in Vaname Shrimp (Litopenaeus vannamei)

Elsa Purwati, Rosa Amalia, Seto Windarto, Sri Rejeki, Dion Saputra

APJMBB 32(4): 97-107

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.11

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Vaname shrimp is an essential commodity in brackish water aquaculture and has high economic value. Infectious diseases are caused by bacteria of the genus Vibrio, one of which is Vibrio parahaemolyticus, and hinder the development of shrimp farming. The increasing bacterial resistance in aquaculture aggravates the situation. To overcome this problem, research into natural materials with new antibacterial activity is needed. Macroalgae are one of the natural materials suitable as a habitat for microorganisms, as they contain various organic substances that serve as nutrients for the growth of bacterial colonies or other microbes such as fungi. Therefore, this study aims to examine the antibacterial activity of endophytic fungi in symbiosis with Caulerpa racemosa against V. parahaemolyticus. C. racemosa from Karimun Jawa Island was isolated in PDA and incubated for seven days. After purification, five isolates showed antibacterial activity using the agar overlay method, and four isolates: C.1.BM, C.4.B, C.7.TM, and C.8.T, showed the best antibacterial activity. The result is that isolates C.1.BM and C.8.T belong to the species Aspergillus fumigatus, isolate C.4.B to the species Aspergillus oryzae, and isolate C.7.TM to the species Trichoderma reesei. The results of this study indicate the potential of the endophytic fungi C. racemosa to be used to prevent disease problems in aquaculture.

Optimization of culture condition of potent strain Phormidium retzii for the optimal production of stigmasterol- an acetylcholinesterase inhibitor using Design of Experiments software

Rincy Yesudas, Vinoth Kumar Thirumalairaj, Geetharamani Durairaj, Amrutha Chacko, Lakshmanasenthil Shanmugaasokan, Suja Gunasekaran

APJMBB 32(4): 108-116

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.12

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Despite the availability of synthetic acetylcholinesterase (AChE) inhibitors for Alzheimer’s Disease (AD), they often possess side effects raising urgent need for natural alternatives providing AChE inhibition with minimal adverse effects. Research has increasingly focused on bioactive products from microalgae, with Phormidium retzii showing promise as an AChE inhibitor, offering novel AD treatment. The research aimed to optimize culture conditions to produce stigmasterol by Phormidium retzii using Design of Experiments (DoE) software. P. retzii was cultivated at pilot scale, with biomass collected, shade-dried, subjected to extraction for AChE inhibition testing until the death phase. The extract containing stigmasterol from 11^th day showed maximum inhibition (81.03±0.23%). Other extracts showed minimal or no inhibition, and the culture reached death phase on 17^th day. Applying N-1 technique, key components of BG11 media for stigmasterol production were defined as sodium nitrate, magnesium sulphate, sodium carbonate and dipotassium hydrogen phosphate. Varying these components in different ratios yielded maximal AChE inhibition (79.43±0.81%) in media containing K₂HPO₄ (0.04 g/L), NaNO₃ (1.5 g/L), MgSO₄ (0.075 g/L) and Na₂CO₃ (0.06 g/L). The optimum light intensity was 2000 lux. Results were input into software, resulting eight experimental runs with ±1 values of each component. AChE inhibitions were determined for all runs and fed into the software. Optimum culture conditions for optimal stigmasterol production were determined as K₂HPO₄ (0.06 g/L), NaNO₃ (2 g/L), MgSO₄ (0.05 g/L) and Na₂CO₃ (0.04 g/L) at pH 8 with 2000 lux. Under these conditions, on 11^th day, dried biomass was 1.22 g with AChE inhibition of 84.36±0.02%.

The effect of lead exposure in preeclampsia: Analysis of superoxide dismutase, homocysteine and MTHFR gene polymorphism

Besari Adi Pramono, Sultana MH Faradz, Suhartono, Hardhono Susanto, Banundari Rachmawati, Noor Pramono

APJMBB 32(4): 117-126

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.13

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Some of the rural areas in Central Java still have the highest maternal mortality rate. Preeclampsia is the leading cause of maternal death in Indonesia. One of the factors contributing to the oxidative stress associated with preeclampsia is lead exposure. Various studies have clearly shown that superoxide dismutase deficiency (SOD) and increased homocysteine (Hcy) levels are consistently associated with the incidence of preeclampsia. Several studies also show the role of MTHFR A1298C and C677T gene polymorphisms in the occurrence of preeclampsia. This study investigated the association between lead exposure, blood SOD and Hcy levels, and MTHFR A1298C and C677T gene polymorphisms in Preeclampsia. This analytical observational case-control study was conducted in 70 cases of preeclampsia and 70 controls. Blood SOD and Hcy levels were measured using Enzyme Linked Immunosorbent Assay (ELISA). Lead level was determined by atomic absorption spectrometry (AAS). MTHFR A1298C and C677T gene polymorphism was genotyped using polymerase chain reaction-restriction fragmentlLength polymorphism (PCR-RFLP). The data obtained were analyzed using the Mann-Whitney U test, chi-square test, independent samples T-test and multivariate linear regression analysis. There was a significant difference (p=0.001) between the lead level in preeclampsia group (mean: 44.50±10.72) compared to the control (mean: 32.78±14.40). The homocysteine level in the preeclampsia cases (mean: 9.06±4.03) also differed significantly (p=0.004) from that of the control group (mean: 7.18±3.09). Increased levels of blood lead and homocystein levels are associated with the incidence of preeclampsia. We found no significant difference in SOD levels, MTHFR A1298C and C677T gene polymorphism between preeclampsia and control group.

Bioinformatic analysis of TFR2, HJV, and HFE nonsense mutations in hereditary hemochromatosis: Insights into the pathogenesis of diabetic mellitus and heart failure

Zahraa Yassen Mohammed, Safa Abbas Khudhair, Tamadher Abbas Rafaa, Ahmed AbdulJabbar Suleiman

APJMBB 32(4): 127-138

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.14

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Sequence variations in TFR2, HJV, and HFE proteins are causative contributors to a wide range of disorders of iron metabolism, which could result in hereditary hemochromatosis, leading to heart failure and diabetes. Known nonsense variants were retrieved from ClinVar, identifying common variants in diabetes mellitus and heart failure, followed by protein-protein interaction analysis through STRING and Cytoscape for hub genes identification, and then structural modeling through AlphaFold and mutation incorporation through DESMOND Maestro to assess their potential impact on protein function. These mutations have not been previously bioinformatically examined, and thus this research presents the foundational studies for further investigations. The modeling and comparative superimposition analysis of wild and mutant proteins showed that nonsense mutations in TFR2 result in truncated proteins that disrupt crucial domains for iron uptake and transport. Similarly, HJV nonsense mutants, including R63* and C321*, truncate the protein and prevent hepcidin regulation. Nonsense mutations in HFE disrupt conserved regions within MHC class I-like antigen recognition and immunoglobulin C1-set domains, potentially leading to abnormal iron buildup. These findings enhance our understanding of the pathogenic mechanisms of genes with these mutations and highlight the importance of genotyping for predicting the severity of iron overload disorders, aiding effective clinical interventions. HFE mutations primarily affect the MHC class I-like antigen recognition domain, while HJV mutations disrupt hepcidin expression and BMP signaling, contributing to juvenile hemochromatosis. Additionally, previously unreported truncating mutations, such as Q12* and R105*, significantly impair function, exacerbating iron overload and disease progression.

Effect of nitrogen ((NH₄)₂SO₄) starvation on growth, lipids, and characterization of fatty acid methyl esther (FAME) in Euglena sp.: Semi-mass cultivation

Eko Agus Suyono, Samia Nashitatuz Zahra, Tariq Akhdan Purnama, Ismia Wulandari, A. Najib Dhiaurahman, Dedy Kurnianto, Khusnul Qonita Maghfiroh, Tia Erfianti, Renata Adaranyssa Egistha Putri, Ria Amelia

APJMBB 32(4): 139-150

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.15

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Microalgae, such as Euglena sp., are microorganisms that can be utilized as biofuel sources. Before mass cultivation, Euglena sp. must adapt through semi-mass cultivation to tolerate various conditions, including environmental changes and contaminants. Nitrogen starvation has been shown to increase lipid production in Euglena sp.; however, the effects of culturing with ((NH₄)₂SO₄) concentrations of 500 mg/L and 250 mg/L have not yet been explored. This research aimed to determine the effects of ((NH₄)₂SO₄) starvation on growth rate and biomass, lipid content, and fatty acid profile of Euglena sp. during semi-mass cultivation (50 L). In this study, nitrogen starvation increased the total unsaturated fatty acids, lipid production, and productivity. However, it reduced cell growth, biomass production, and productivity. The highest lipid productivity was observed at 500 mg/L ((NH₄)₂SO₄), reaching 0.011±0.0014 mg/mL and 0.217±0.014 mg/mL for lipid production on day 15. Meanwhile, the highest biomass production occurred at 1000 mg/L ((NH₄)₂SO₄), reaching 0.156±0.010 mg/mL on day 15 and a biomass productivity of 0.0067 mg/mL. Methyl palmitoleate (C16:1) was the dominant FAME in all treatments, reaching 18.38% at 500 mg/L ((NH₄)₂SO₄). Thus, it can be concluded that the 500 mg/L ((NH₄)₂SO₄) treatment is more efficient in increasing FAME, lipid production, and productivity for biofuel production.

Molecular typing of multidrug resistant Klebsiella pneumoniae recovered from Iraqi burned patients

Mustafa Mohammed Touma, Bassam K. Kudhair, Karrar S. Zayed, Inam J. Lafta

APJMBB 32(4): 151-159

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.16

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Klebsiella pneumoniae causes lethal nosocomial infections, mostly affecting patients with severe burns. More than 80% of its isolates have shown resistance to routinely used antibiotics in parallel with increased infection rates. The study aimed to determine the molecular typing and genetic relatedness of K. pneumoniae. Therefore, 20 multidrug resistant (MDR) K. pneumoniae already isolated from infected burned wounds in two major hospitals of Al-Kut city east Iraq were subjected to genotyping analysis. The random amplified polymorphic DNA (RAPD)-based polymerase chain reaction (PCR) technique was used along with three oligonucleotide primers (P13, OPX-04, and OPY-01). The amplicons’ patterns of the electrophoresis-gel were analyzed by the GelJ software. Results revealed various patterns of DNA bands. A genetic similarity was seen within isolates from some locations. This genetic relatedness was captured by dendrogram analysis of the generated RAPD profiles. However, a genetic diversity among K. pneumoniae clinical isolates was also detected suggesting their different origins as well as ongoing changes of the bacterial genome. Furthermore, this could propose the circulation of many strains simultaneously within the hospitals. Therefore, it is important to consider this genetic heterogeneity when developing control measures for nosocomial K. pneumoniae infections. In conclusion, the current study highlights the dissemination of various MDR K. pneumoniae strains in the burn wards of two major hospitals in Al-Kut city, Iraq. Similar studies need to be performed in other Iraqi hospitals to establish a data base used in infection control systems, to predict, and manage the spread/outbreaks correlated with certain genotypes of resistant strains.

Bacteria and viruses in gene therapy: An evidence-based review

Firyal Ayyal Musa, Dunia Kamal Salim, Dhamyaa Obaid Shalgam, Khaldoon Jasim Mohammed

APJMBB 32(4): 160-171

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.17

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Gene therapy means introducing genetic information into a cell to treat or prevent disease. It can replace defective genes, suppress harmful genes, or enhance cellular functions; therefore, it is considered promising for various diseases such as genetic disorders, cancer, and viral infections. This review assembled evidence related to bacterial (Salmonella typhi, Escherichia coli, Listeria monocytogenes, and Lactococcus lactis) and viral (Retroviruses, Herpes simplex viruses, Lentiviruses, and Adenoviruses) vector-mediated gene therapeutics along with their efficacy, safety, and possible uses in gene therapy. The results demonstrated bacterial vectors can transfer their genes, especially in cancer treatment. Research has shown that live Salmonella strains can preferentially home into tumors and suppress their growth. E. coli has been modified to enhance the ability to transfer genetic material and minimize toxic impacts. Listeria monocytogenes bacterium has been considered for cancer treatment through immunotherapy, while Lactococcus lactis has the potential for use in inflammatory diseases because of its probiotic qualities. Surprisingly, viral vectors continue to dominate the field of gene therapy because they are effective in transferring genes. Both retroviruses and lentiviruses have been employed due to their capacity to integrate ad hoc within the host cell genome and maintain gene expression over long periods. Human herpes simplex viruses exhibit significant packaging capacity and neurotropism, while adenoviruses are utilized effectively in various cancer treatment applications.

Preparation and evaluation of antioxidant activity of Spirulina platensis extraction: Potential application in cancer prevention and treatment

Azhaar Asker Hamadi, Ali Noory Fajer

APJMBB 32(4): 172-181

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.18

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Spirulina platensis, a type of blue-green algae, is renowned for its bioactive compounds that act as antioxidants. These compounds have been found to mitigate or prevent oxidative stress caused by free radicals, which are implicated in the development of cancer. In this study, we prepared aqueous, hexane, and alcohol extracts of Spirulina platensis and assessed their antioxidant activity using the DPPH method. Additionally, gas chromatography-mass spectrometry (GC-MS) was applied to recognize the chemical components of the Spirulina extractions. The results indicated that the alcoholic extraction showed higher antioxidant activity (49.9%) compared to the aqueous (34.2%) and hexane (11.1%) extractions. The study's findings suggest that extracts of Spirulina platensis contain a significant amount of bioactive compounds, which may contribute to its strong anticancer and antioxidant properties. These findings suggest the potential for developing new anticancer agents derived from blue-green algae.

Acacia arabica-enriched zinc oxide biopolymer nanocomposites: A dual-purpose strategy for combatting MCF-7 human breast cancer cells

Sarojini Savarimuthu, Dinesh Karthik Arunachalam, Mary Nancy Flora Rayappan, Geetha Kannapan, Palani Subramani

APJMBB 32(4): 182-194

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.19

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In the global effort to combat breast cancer, this study investigates the potential of Zinc Oxide Biopolymer Nanocomposites (ZnO-BNCs) as a multimodal treatment and diagnostic option for breast cancer, specifically targeting the MCF-7 cell line. ZnO-BNCs were synthesized using environmentally friendly Acacia Arabica leaf extract, known for its arabinoxylan biopolymer, which enhances the biocompatibility and stability of the nanocomposites. By combining the anticancer properties of ZnO nanoparticles with the biopolymer matrix, ZnO-BNCs demonstrated improved therapeutic efficacy and targeted delivery. Comprehensive characterization techniques, including surface morphology analysis, particle size determination, stability assessment, and elemental composition analysis, were employed to assess the properties of the ZnO-BNCs. The Box-Behnken design was used to optimize the synthesis process, showing promising results. The cytotoxicity evaluation using the MTT assay revealed a significant reduction in MCF-7 cell viability, with a 13-fold decrease at the highest concentration (500 μg/mL) compared to the control. These findings suggest that ZnO-BNCs hold great potential as a functional biomaterial for future biomedical applications, particularly in cancer treatment and diagnosis.

Hormonal and metabolic profiles and their association with BMI in Iraqi women with Polycystic Ovary Syndrome

Reem Husam Al-Tabra

APJMBB 32(4): 195-201

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.20

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Polycystic ovary syndrome (PCOS) is a problem with hormones that occurs during the reproductive years. If you have PCOS, you may not have your period very often. Or you may have periods that last many days. You may also have too much of a hormone called androgen in your body. This study included a sample of 35 Iraqi women who had been diagnosed with polycystic ovary syndrome (PCOS), and a control group of 35 healthy individuals. Anthropometric measurements, such as body mass index (BMI) and waist-to-hip ratio (WHR), were documented. Hormonal and metabolic profiles were examined in blood samples from all participants. The statistical analysis system (SPSS) version 28 was used to calculate mean ± SD and p-value. LH, LH/FSH ratio and BMI levels were significantly higher in obese PCOS patients compared to control subjects. In addition, the study showed an LH/FSH ratio level of 0.5774±0.9221 for the control groups and 1.0061±0.310 for the POCS groups. The insulin level was 7.874±0.6978 for the control groups and 27.07±6.354 for the POCS groups. The estrogen level was 58.62 ± 27.32 for the control groups and 78.20 ± 32.4 for the POCS group. All the results of LH/FSH ratio, progesterone, prolactin, testosterone, estrogen, insulin, and Zn were significantly higher in obese PCOS compared to the control group. Although adropin contributes to better health regulation compared to women with PCOS, its impact was less pronounced in women with PCOS.

Functional analysis of the antisense long noncoding RNA ANO1-AS2 and the ANO1 gene in Iraqi men with idiopathic infertility

Shimaa Jumaa Abood, Maan Hasan Salih, Mohammad Mutlag Salih

APJMBB 32(4): 202-209

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.21

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The leading causes of male infertility remain unclear. Men with average semen analysis results are diagnosed with idiopathic infertility. The ANO1-AS2 gene is a long noncoding RNA located close to the anoctamin1 (ANO1) gene. ANO1 regulates calcium-activated chloride channels, which regulate many cellular processes. In this study, we aimed to evaluate the relationships between ANO1-AS2 and ANO1 expression in men with idiopathic infertility. ANO1-AS2 and ANO1 expression was evaluated in sperm samples from 50 men with idiopathic infertility and 50 fertile men. Quantitative real-time polymerase chain reaction was used to analyse the sperm samples. The results showed that ANO1 expression was significantly higher than ANO1-AS2 expression (p = 0.0001). Moreover, there was a positive correlation between ANO1 mRNA levels and sperm motility (p < 0.001; r = 0.7189). Conversely, a significant negative correlation was found between ANO1-AS2 levels and sperm motility (p < 0.003; r = -0.4068). ANO1 mRNA levels were also negatively correlated with abnormal sperm morphology (p < 0.0137; r = -0.3464), whereas ANO1-AS2 levels were positively correlated with abnormal sperm morphology (p < 0.0127; r = 0.350). The observed inverse relationship between ANO1-AS2 and ANO1 expression indicated that ANO1-AS2 may play a role in ANO1 downregulation. However, the expression of both genes was not related to the cause of idiopathic infertility in men in average semen analysis. Further studies are needed to determine the potential correlation between male infertility and ANO1-AS2 in other ethnic groups and types of infertility.

Enhancing carbohydrate binding module family 40 (CBM40) via in silico sequential mutagenesis approach

Nazmi Harith-Fadzilah, Nadiawati Alias

APJMBB 32(4): 210-221

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.22

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The Carbohydrate-Binding Module family 40 (CBM40) is a component of bacterial sialidase that exhibits a high affinity and selectivity for sialic acids. These CBM40s can be independently isolated and modified for use as drug delivery agents against tumours exhibiting hypersialation. Modifying specific amino acid sequences can enhance the sialic acid binding ability of CBM40. This study employed an in silico approach to construct, simulate and evaluate Vibrio cholerae CBM40 mutants via interaction of these mutants with a sialoside ligand via molecular docking and molecular dynamics simulation. Several rounds of mutagenesis were proposed to sequentially improve the CBM40 ligand binding energy. There were discprepancies between the molecular docking and simulation data from the first round of mutagenesis. The CBM40 residue substitution from Arg74 to Trp74 and from Gly196 to Gln196 (R1-Gly-Gln196) had an equal molecular docking binding energy increase. However, the former mutant exhibited weaker binding than non-modified CBM40, while the latter displayed stronger ligand binding. Two mutants identified in the second round of mutagenesis exhibited improved ligand binding energy from the molecular docking analysis. However, both mutants displayed weaker free binding energy than the R1-Gly-Gln196. Molecular dynamics simulation post-analysis suggested R1-Gly-Gln196 formed a more stable interaction with sialoside compared to other CBM40s. The analyses also revealed that mutation of residues can directly or indirectly introduce steric clashes which cause interaction instability and thus reduces ligand binding energy. This study highlighted the significance of validating molecular docking results with simulation analysis, thereby improving the reliability of in silico protein-ligand interaction predictions.

The bioactive benefits of Chenopodium quinoa crude protein: From antibacterial to anticancer properties

Amit Sen, Nalini Tomer, Hemlata Srivastava, Manas Mathur, Sahaya Shibu B., Sarmad Moin

APJMBB 32(4): 222-230

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.23

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This study explores the multifaceted attributes of quinoa crude protein, shedding light on its potential applications in various fields. Quinoa is a crop with great potential because of its high nutritious contents and is the subject of growing interest due to its potential health benefits. This research aims to uncover the properties and applications of quinoa crude protein, including its protein content, antimicrobial potential, protease activity, and anticancer potential. Extracted crude protein from Quinoa seed was estimated using the Folin Lowry method, and SDS-PAGE to identify distinct protein profiles. The antimicrobial activity of the crude protein was assessed through Minimum Inhibitory Concentration (MIC). Protease activity was measured, and the cytotoxicity of the crude protein was determined on various cell lines. The study found that quinoa crude protein contains 0.354 mg/gm of protein, though this may vary based on quinoa variety and growth conditions. SDS-PAGE analysis identified four prominent proteins with minimal disruption during extraction. The protein exhibited strong antimicrobial potential, with low MIC values against specific bacterial strains. The quinoa crude hydrolysate displayed significant protease activity, indicating potential industrial applications. The cytotoxicity of the protein was dose-dependent, with a notable impact on cancer cells. Quinoa crude protein holds promise for diverse applications. Its variable protein content, antimicrobial potential, protease activity, and cytotoxic effects on cancer cells all suggest its potential in fields ranging from nutrition and health to biotechnology and industry. 

Acidified water extraction increases carpaine yield from Carica papaya L. Leaf

Sim Tze Tak, Nor Hisam Zamakshshari, Mohamad Fhaizal Mohamad Bukhori, Eswaran Madiahlagan, Rafeah Wahi, Lawrence Anchah

APJMBB 32(4): 231-244

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.24

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Carica papaya leaves have been traditionally used in various medicinal applications. The major alkaloid, carpaine, is widely studied for its anti-thrombocytopenic activity. However, extraction of carpaine from Carica papaya leaves remains challenging, and the existing methods may not yield high amount of carpaine due to the presence of various interfering compounds. High carpaine yields are essential for studying on its anti-thrombocytopenic and immunomodulatory effects, assessing its efficacy and safety as a potential malaria drug candidate. This study aims to extract and isolate carpaine from Carica papaya leaves using alcohol extraction method and acidified water extraction method performed on dried powdered leaves and compare their percentage yield of carpaine. The percentage yield of carpaine isolated from acidified water extraction was 12.01%, while the yield for alcohol extraction was 2.18%. Acidic condition enhances carpaine's water solubility through protonation, converting carpaine in its free base form to salt form which improves extraction efficiency. Heat and stirring further optimize the acidified water extraction process. Characterization of isolated carpaine was performed with various analytical techniques to confirm the structure of carpaine isolated.

microRNA-122 gene expression in Iraqi patients with hepatocellular carcinoma

Rana Talib Mohsen, Al-Taee Haneen Z, Wafaa Hussien Habeeb, Riyadh A. Abd-Alazeez, Sally Hassan, Shahlaa Talib Mohsen

APJMBB 32(4): 245-250

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4.25

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Hepatocellular carcinoma (HCC) is one of the most prevalent tumors worldwide. It is estimated that persistent infections with hepatitis B (HBV) or hepatitis C (HCV) viruses account for 80% of HCC cases. Both chronic HBV and HCV infections have the potential to modify hepatocyte function and may do so through comparable pathways that impact the development of HCC. Significant progress has been made in understanding of the molecular biology of HBV and HCV and the cellular signal transduction pathways affected by these illnesses. Several cancer types exhibit aberrant expression and deregulation of microRNA-122, suggesting these molecules might function as prognostic and diagnostic biomarkers globally. For detection of HCC, non-invasive biomarkers are limited in their application. In order to enhance the survival rates of individuals with HCC, early identification is crucial. Quantitative RT-PCR was used to evaluate 64 serum samples. The levels of microRNA-122 in the serum of HCC patients were significantly lower than in the control group further contrasted the expression of these microRNAs in serum samples from healthy controls and patients with untreated early-stage HCC. MicroRNA-122 expression was considerably reduced in HCC sera, with a P value of less than 0.0001. Interestingly, microRNA-122 expression levels were marginally increased in individuals who received therapy but were lacking in those diagnosed early and untreated. In the first phase of hepatitis samples, microRNA-122 expression levels were considerably lower than in standard samples, with a P value (<0.0001). This suggests the potential use of microRNA-122 as a biomarker to track treatment success.

Volume 32(4) (Special Issue); 2024

Biodegradation of low-density polyethylene (LDPE) by Aspergillus niger and Aspergillus oryzae 

Mohamad Syafiq Zulkafli, Fahim Rithwan, Dayang Norulfairuz Abang Zaidel, Solleh Ramli, Siti Zulaiha Hanapi, Nur Izyan Wan Azelee, Tong Woei Yenn, Hesham A. El Enshasy, Daniel Joe Dailin

APJMBB 32(4) (Special): 1-10

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).01

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Plastic waste, particularly polythene, is a major environmental concern, with bioremediation emerging as a sustainable solution. This study evaluated the potential of Aspergillus niger and Aspergillus oryzae in degrading low-density polyethylene (LDPE). Fungal species were initially cultivated on potato dextrose agar (PDA) plates at 36°C for 5 days in the dark. Subsequently, they were transferred to a minimal salt medium supplemented with LDPE strips. The degradation efficiency was evaluated by measuring the weight loss of the LDPE samples and conducting scanning electron microscopy (SEM) analysis. The results revealed weight loss ranging from 1.50 to 10.22%, depending on the fungi and incubation period, whereas control samples without fungal exposure showed no change. Scanning electron microscopy (SEM) images confirmed fungal colonisation and physical degradation of the LDPE surface, displaying folds, grooves, and cracks. Results from this study showed that both fungi were able to break down the plastic, with A. oryzae showing more significant degradation. These findings highlight the potential of fungi as a biological solution to address the global plastic pollution issue.

In vitro modulatory effect of nanoencapsulated tocotrienol-rich fraction, ascorbyl tetraisopalmitate, and carotenes on atopic dermatitis-like model of human epidermal keratinocytes 

Yee-Lin Gan, Chin-Ping Tan, Yoke-Kqueen Cheah, Hidayah Ariffina, Helmi Wasoh, Oi-Ming Lai

APJMBB 32(4) (Special): 11-26

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).02

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The aim of the study was to investigate the regulatory effects of mixed antioxidants, including tocotrienol-rich fraction, ascorbyl tetraisopalmitate, and carotenes, on inflammatory response and epidermal integrity of normal human epidermal keratinocytes (NHEK) stimulated with cytokines to mimic atopic dermatitis (AD). Nano encapsulated mixed antioxidants (NMA) were prepared by encapsulating tocotrienol-rich fraction, ascorbyl tetraisopalmitate and carotenes into nanoemulsion. The interleukin (IL)-4 (50 ng/mL), interferon (IFN)-γ (50 ng/mL), and tumor necrosis factor (TNF)-α (20 ng/mL) individually stimulated NHEK cells were treated with NMA (2.1-2100 ng/mL). Then, cytokines, chemokines and structural proteins expressions were determined. Cell monolayer integrity was assessed using dispase-based keratinocyte dissociation and wound-healing assays.The key findings of the study showed that NMA treatment (2100 ng/mL) resulted in a complete abolishment of upmodulation thymic stromal lymphopoietin (TSLP) expression induced by IL-4, IFN-γ, and TNF-α. Upregulated expression of thymus and activation regulated chemokine (TARC) following IL-4 stimulation was attenuated by NMA (2.1-2100 ng/mL). The increment of interferon gamma-induced protein 10 (IP-10) and monocyte chemoattractant protein-1 (MCP-1) expressions resulting from IL-4, IFN-γ, and TNF-α stimulations was significantly suppressed by NMA treatment. The downregulated filaggrin expression provoked by IL-4 and TNF-α stimulations was abrogated with NMA treatment (2.1-2100 ng/mL). Treatment with NMA significantly improved the production of cornified envelope proteins, decreased the dissociation scores and accelerated wound healing of IL-4, IFN-γ, and TNF-α stimulated NHEK.

Sequence analysis and structural modelling of major capsid protein L1 of avian papillomavirus from African Grey Parrot

Zacharia Kadiayeno Egbunu, Yong Zi Yap, Nurulhuda Najihah, Abdul Razak Mariatulqabtiah

APJMBB 32(4) (Special): 27-37

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).03

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Papillomaviruses are non-enveloped, and icosahedral in structure with a double-stranded circular DNA. They are responsible for inducing regressing papillomas (warts) on mucosal or keratinized epithelia across a diverse range of species including mammals, reptiles, birds and fish. Unlike human and bovine papillomaviruses, avian papillomaviruses (AvPV) received little attention in terms of sequence analysis and protein structure repository. This may be due to the less severity of morbidity and mortality compared to papillomavirus disease manifestations in human. The African Grey Parrot (Psittacus erithacus) is the first avian species to have a complete AvPV genome sequenced (PePV). Nonetheless, sequence analysis of its genes is limited with no three-dimensional structure reported in Protein Data Bank. Therefore, the aims of this study are to analyse the sequence of major capsid protein L1 of PePV, to assess its physicochemical properties, to generate its secondary and three-dimensional structures and to elucidate the quality of the generated L1 structural models. The PePV L1 capsid protein was analysed using online bioinformatics tools namely NCBI GenBank, PaVE, MUSCLE, ProtParam, PSIPRED and SOPMA.  SWISS-MODEL, RaptorX and C-I-TASSER were implemented for high-quality structural modelling prior to comparison using PyMOL and Molprobity. Results demonstrated that the PePV L1 capsid protein was slightly acidic and thermally-stable. Additionally, the PePV’s host specificity is closed related to Fringilla coelebs papillomavirus (FcPV1) and Serinus canaria papillomavirus (ScPV1), both of which infect birds from the Austravales clade. Structure predictions reveal slight structural difference and similarities albeit the SWIS-MODEL and C-I-TASSER showed relatively high-quality models which were considered as the basis for structural comparison and reliability. Further research on avian major capsid protein L1 of PePV is anticipated to improve the current knowledge on AvPV’s structure-function relationship thus control the viral transmission in endangered birds.

Enhancing polyhydroxalkanoate (PHA) production from phenol through fermentation strategies: A review

Izzati Sabri, Mohd Zulkhairi Mohd Yusoff, Nor Azlan Nor Muhammad, Li Sim Ho, Norhayati Ramli

APJMBB 32(4) (Special): 38-50

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).04

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Microbial conversion offers a promising solution to two environmental challenges, phenol and plastic pollutions, via the transformation of phenol into bioplastics, specifically polyhydroxyalkanoate (PHA). Synthetic plastics are widely used across various sectors; however, their non-biodegradable nature and extensive daily use significantly contribute to environmental deterioration. Similarly, phenol, an important industrial material, is often released into the environment through inadequately treated effluents. Phenol is toxic even at low concentrations and may lead to severe environmental and health problems if not properly managed. Microorganisms not only degrade phenol into non-harmful compounds, facilitating its removal from the environment, but they also accumulate intracellular PHA, providing a biodegradable alternative to synthetic plastics. However, phenol’s toxicity at high concentrations can inhibit this process, leading to cell death. This review explores various fermentation strategies aimed at enhancing PHA production while addressing phenol toxicity. These strategies include the use of mixed microbial community (MMC), acclimatization to increasing phenol concentrations, feast-and-famine strategies, co-substrate supplementation, and substrate feeding strategies. An integrated approach would be more effective in overcoming phenol toxicity, leading to complete phenol degradation and improved PHA accumulation. However, these strategies must be tailored to the capabilities of microorganisms in adapting to and utilizing phenol as feedstock. Overall, these fermentation strategies have the potential to improve the management of plastic waste and phenol-contaminated wastewater, contributing to a more sustainable future.

Chemical oxygen demand reduction in wastewater by locally isolated Priestia sp. BA01 strain

Gao Jianfeng, Murni Halim, Mohd Shamzi Mohamed, Rosfarizan Mohamad

APJMBB 32(4) (Special): 51-64

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).05

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Chemical oxygen demand (COD) refers to the oxidant content consumed by treating water samples with a certain strong oxidant under certain conditions, which is expressed in mg/L. This index is widely used in the world to reflect the degree of organic pollution in water. Using heterotrophic bacteria such as Bacillus sp. to reduce COD value in wastewater has become a common technology. The isolation and study of efficient and stable strains has great significance. In this study, a bacterial strain with COD reduction ability was isolated from the shallow soil of UPM campus. The physiological and biochemical study, and 16srRNA gene identification were carried out. The cultivation and application conditions were optimized. It was confirmed that the isolated BA01 strain belongs to Priestia sp. and the sequence has been submitted to NCBI GenBank. The GenBank number is PQ002608. For the cultivation conditions optimization, OFAT and orthogonal test methods were used. The best conditions are at 8 h, 35 ℃, 250 rpm and pH 7.0, which the biomass reached at 2.32×10¹⁰ CFU/ml. The pH value and rotation rate are significant. For application conditions optimization, OFAT and RSM were used. The best application conditions are at 35.22℃, pH 7.09, 215.45 rpm. Under these conditions, the COD reduction rate is 63.32%, 189.96mg/L. In addition, pH value is significant whereas the pairwise relationships are insignificant. Orthogonal experiments and RSM both aid in the systematic analysis of experimental results. Orthogonal experiments significantly reduce workload, but they have certain limitations in terms of interactions between factors. RSM can capture interactions and nonlinear relationships between factors, but it needs a greater number of tests than orthogonal experiments.

Emergence of extended spectrum beta-lactamases and carbapenemases producing Klebsiella spp. isolated from Hospital Sultan Abdul Aziz Shah

Vakgesri Muniandy, Sharleen Livina Isaac, Syafinaz Amin Nordin, Adelene Ai-Lian Song, Amalia Mohd Hashim, Wan Nur Ismah Wan Ahmad Kamil

APJMBB 32(4) (Special): 65-77

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).06

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The emergence of extended spectrum β-lactamases (ESBLs) and carbapenemases producing Klebsiella spp. in hospital settings is alarming as it leads to resistance to carbapenems, the last resort of antibiotics. Therefore, the objective of this study is to determine the antibiogram profile and identify the resistance genes present in Klebsiella spp. clinical isolates, encompassing specimens such as urine, sputum, blood and pus. The presence of β-lactamase genes; blaTEM-1 and blaCTX-M-1 and carbapenemases genes; blaNDM-1 and blaVIM- 1 were detected by polymerase chain reaction (PCR) analysis. A total of 85 isolates were collected from ill patients at Hospital Sultan Abdul Aziz Shah (HSAAS) between January 2022 and March 2023. The resistance profile of these isolates was analysed using the Kirby-Bauer disk diffusion method against various classes of antibiotics. The results revealed that all 85 Klebsiella spp. tested were resistant to ampicillin and ciprofloxacin. Furthermore, 78% of isolates were resistant to cephalosporins, suggesting that these strains were producing ESBLs. In terms of carbapenem resistance, the isolates were more resistant towards meropenem (80%) than imipenem (60%). These 85 clinical isolates were also resistant to kanamycin (52%). The presence of blaTEM-1 and blaCTX-M-1 genes were detected in the tested isolates, namely strain number 15,18,27,64 and 65. Moreover, the carbapenem-non- susceptible isolates were detected to have blaNDM-1 and blaVIM-1 genes. These findings highlighted a substantial proportion of isolates as ESBLs and carbapenemase producers. In conclusion, this study emphasised the urgent need for enhanced surveillance programs to combat the escalating threats of multidrug resistance Klebsiella spp. in clinical settings. Additionally, routine molecular screening for resistance genes along with rapid diagnostics for detecting these genes, should be implemented to enable doctors to prescribe the correct antibiotic.

Cost-effective drying methods for edible bird’s nest hydrolysate:
Impact on nitrite levels and sialic acid content

Bee-Hui Yeo, Mithula A/P Anamaly, Oi-Ming Lai

APJMBB 32(4) (Special): 78-93

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).07

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To produce powdered edible bird’s nest hydrolysate (EBNH), a significant amount of water must be removed after hydrolysis to improve the product’s stability, extend shelf life, and reduce transportation and storage costs. This study investigates the application of cool-air and hot-air drying methods, with a particular focus on strategies suited for small and medium-sized enterprises (SMEs). Cool-air drying was performed at 17 °C using a fan and air conditioner, while hot-air drying was conducted at 55 °C and 65 °C using a dehydrator. The study evaluated drying time, as well as the quality, moisture content, nitrite levels, total sialic acid content, and antioxidant activity of EBNH produced by each method. The quality, cost, and feasibility of these methods were compared with freeze-drying and spray-drying techniques from previous studies. Results showed that higher temperatures and lower relative humidity improved drying efficiency, with hot-air drying at 65 °C requiring the shortest drying time. Chemical analysis revealed significant differences in nitrite and total sialic acid content: cold-air-dried samples had higher nitrite levels, while hot-air drying at 65 °C produced the highest total sialic acid content. Cost and feasibility assessments demonstrated that hot airdrying at 65 °C was the most efficient and cost-effective method. This method is especially advantageous for SMEs due to its balance of efficiency, affordability, and ease of implementation, making it ideal for smaller-scale operations.

Complete genome sequence of local Malaysian Fowl adenovirus serotype 8b UPM T221 strain

Bahiyah Azli, Nur Farhana Salim, Mohd Hair-Bejo, Norfitriah Mohamed Sohaimi, Nor Asilah Wati Abdul Hamid, Nurulfiza Mat Isa

APJMBB 32(4) (Special): 94-113

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).08

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Recently, Fowl adenovirus serotype 8b (FAdV-8b) infection has emerged as a serious threat to Malaysia’s poultry industry, acting as a causal agent of Inclusion Body Hepatitis (IBH) with a mortality rate of 10-30% among chickens. In this study, we isolated an FAdV strain from the liver of an IBH-positive dead commercial broiler chicken in Tawau, Sabah, and subjected it to pathogenicity analysis and Whole genome sequencing (WGS). Upon inoculation of UPM T221 isolate into Specific pathogen-free chicken embryonated eggs, slow mortality pattern of 6- to 12-days post-infection (dpi) was recorded, with nil gross lesions of both chorioallantoic membrane (CAM) and liver observed during harvesting. Upon subjection to WGS, the genome of UPM T221 was found to be 44722 bp in length with 58.1% GC content, 37 coding sequence (CDS), identifying the isolate as a strain from FAdV-8b of the FAdV-E. Interestingly, the left ORF regions and central genes showed a higher potential for significant genetic divergence with Single nucleotide polymorphism (SNPs) and InDels gaps. These variants are displayed in the within the structural capsid of UPM T221, such as hexon and penton, as well as in virion replication processes such as DNA polymerase and pTP. These findings of less virulence, non-pathogenic UPM T221 profile proposed the strain as a suitable live attenuated candidate vaccine, also contributing to the current understanding of the genetic diversity of FAdV for developing autogenous vaccines or diagnostic materials especially against local strain infections.

Evaluation of Misai Kucing (Orthosiphon stamineus) extract on diabetic cell line

Adieya Atyrrah Adnan, Mohd Ezuan Khayat, Murni Halim, Helmi Wasoh, Zulfazli M. Sobri

APJMBB 32(4) (Special): 114-126

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).09

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Orthosiphon stamineus, locally known as Misai Kucing, is a traditional medicinal plant used to treat various ailments, including diabetes. However, limited scientific research has confirmed its anti-diabetic potential. This study aimed to evaluate the effects of O. stamineus extracts on diabetic cell lines. The plant was dried, ground, and extracted using four different solvents: water, ethanol, ethyl acetate, and hexane. The crude yield from these extractions was highest in water, followed by ethanol, ethyl acetate, and hexane. In vitro toxicity assays were conducted on the 3T3-L1 pre-adipocyte cell line to assess the viability of cells treated with water and ethanolic extracts. The extracts were tested at various concentrations to determine their toxicity. Concentrations below 1.25 x 10^-1 mg/mL for both extracts were non-toxic, with more than 50% viable cells observed. However, at higher concentrations (1.25 x 10^-1 mg/mL), the water extract showed toxicity. Non-toxic concentrations (ranging from 0 to 0.06 mg/mL) were used to measure glucose uptake in the 3T3-L1 diabetic cell line by incubating the cells with a fluorescent D-glucose analog, 2-NBDG. The results showed a reduction in 2-NBDG uptake in cells treated with the extracts compared to untreated cells, indicating the potential to influence glucose metabolism. This suggests that water and ethanolic extracts of O. stamineus could play a role in managing diabetes by improving glucose regulation, supporting its traditional use as an anti-diabetic remedy and highlighting its medicinal potential.

Optimisation of casein adsorption onto alkaline-treated rice husk using response surface methodology for sustainable protein recovery

Angelina Phoebe Vincent, Mohd Badrin Hanizam Abdul Rahim, Mohd Yunus Abdul Shukor, Mohd Ezuan Khayat

APJMBB 32(4) (Special): 127-134

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).10

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With rising environmental concerns and a need for sustainable resource management, protein recovery from dairy wastewater has gained significant attention. This study investigates the use of alkaline-treated rice husk as an adsorbent for casein recovery, offering a low-cost and eco-friendly alternative for waste valorization. Using response surface methodology (RSM) and a Box-Behnken Design (BBD), the effects of contact time, adsorbent dosage, and pH on adsorption capacity were systematically evaluated. Optimal conditions were determined as 30 minutes of contact time, 4 mg/mL adsorbent dosage, and pH 9.93, achieving a predicted adsorption capacity of 111.547 mg/g. Validation experiments confirmed the model’s accuracy, yielding an actual adsorption capacity of 110.0 mg/g, with no significant difference from the predicted values (p > 0.05). These findings demonstrate the potential of alkaline-treated rice husk for efficient protein recovery, contributing to sustainable nutrient recycling and environmental protection. This optimized adsorption process holds promise for industrial applications in waste management within the dairy sector, supporting resource conservation and sustainable practices. However, the study is limited by the lack of long-term stability testing of the adsorbent and its performance in real industrial wastewater, which should be addressed in the future.

Equilibrium, kinetics, and thermodynamic studies on the biosorption of Reactive Red 120 dye utilizing the biomass of Enterobacter sp. MMO5

Salihu Yahuza, Motharasan Manogaran, Nur Adeela Yasid, Ahmad Razi Othman, Mohd Yunus Abd Shukor

APJMBB 32(4) (Special): 135-163

Article DOI: https://doi.org/10.35118/apjmbb.2024.032.4(Special).11

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Biosorption using sustainable biomass such as bacteria is highly desirable due to their large surface area. Enterobacter sp. MM05 was the organism of choice for the Reactive red-120 dye biosorption. The dye sorption optimization was done using Response Surface Methodology (RSM) and One-Factor-At-Time (OFAT). Upon RSM optimization, 50 mg/L, 150 rpm, 60 min, 7.0, and 45^oC were the optimum results for the concentration, agitation, time, pH, and temperature, respectively. The biosorbent was characterized using Fourier Transform Infrared (FTIR) and Scanning Electron Microscopy (SEM) analyses. Biosorption isotherms, kinetics, and thermodynamic parameters were studied using nonlinear regression. Compared to the traditional One-factor-at-a-time (OFAT) method, the adsorption rate was about 6% higher after optimization using response surface methodology via Central Composite Design (CCD). The pseudo-second-order kinetics reaction fitted the dye biosorption with the lowest AICc and highest adjR² values. Langmuir, Freundlich, Henry, BET, Sips, Toth, Fritz-Schlunder IV, and Fritz-Schlunder V were the mathematical isotherm models with the best fit. Except for Henry, all the isotherm models tested on the RR-120 dye provided significant fitting results. Freundlich isotherm was the best after statistical analysis, having the lowest AICc value of -51.54. The thermodynamic parameters were computed using non-linear regression based on the vant Hoff plot. The enthalpy change (ΔH°) value was 52.91 kJ/mol, indicating that the reaction was endothermic. The adsorption process was spontaneous and thermodynamically feasible, as denoted by the negative values of Gibbs free energy (ΔG°) calculated at various temperatures. An increase in the degree of randomness at the solid/liquid interface was indicated by the positive entropy change (ΔS°), which is likely due to the structural changes in the bacterial biomass and Reactive Red 120 dye upon binding. This study demonstrated the potential of the bacterial biomass as a good biosorbent for dye biosorption, particularly Reactive Red 120, and offers a promising alternative for the bioremediation of textile dyes.