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Fulltext Identification, characterisation and phylogenetic analysis of commensal bacteria isolated from human breast milk in Malaysia

Zubaida Hassan, Shuhaimi Mustafa, Raha Abdul Rahim, Nurulfiza Mat Isa

Pertanika Journal of Science & Technology, 2016;24(2):351-370.
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Human breast milk microbiota is essential for infant immune system development, maturation and
protection against infection. However, there is scarce information on the fluid’s microbiological
composition from Malaysia. The objective of the study was to isolate, identify and characterise commensal
bacterial population present in human breast milk from Malaysia. One hundred bacteria were isolated
from the human breast milk of healthy lactating women (n=30). After preliminary screening, 20 isolates
were characterised using both phenotypic and molecular techniques. The results indicated that most
frequently identified bacteria in this study were E. faecalis and S. hominis. These organisms alongside E.
cloacae were all metabolised D-Maltose, Sucrose, D-Turanose, α-D-Glucose, D-Fructose, D-Mannose,
D-Galactose, D-sorbitol and D-Mannitol and were able to grow at pH 5 and 6, 1% sodium lactate, 1%,
2% and 8% NaCl. BLAST showed over 99% similarity to those deposited in Genbank. Phylogeneticrelatedness
was depicted using neighbour-joining method and had two clades with 100% bootstrap. These
findings provided insight into the nature, characteristics and also phylogenetic-relatedness of bacteria
present in human milk from Malaysia. Isolation and identification of commensal bacteria from human
milk are considered the first step for future studies on the benefit of these organisms towards human health.
Fulltext Determining the efficacy of CRISPR-Cas9 mechanisms on the uppP Gene of Klebsiella pneumonia using Lysozyme assay

Mohd Nasharudin Razak, Raha Abdul Rahim, Abdul Rahman Omar, Zunita Zakaria, Nurulfiza Mat Isa

Malaysian Journal of Medicine and Health Sciences, 2020;16(4):226-233.
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Introduction: Multidrug resistance bacteria is alarming worldwide. A lot of research were done and are ongoing to search for the best, convenient and economically affordable ways to fight them. With the latest genome editing tool; Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology, this research was performed to develop a novel strategy to genetically modify the genome and inhibit the growth of Klebsiella pneumoniae (UPM ESBLKP1), an Extended Spectrum Beta Lactamases (ESBL) organism. Methods: A CRISPR-Cas9 vector was construct- ed together with guide RNAs designed specifically for the targeted uppP gene, a gene responsible for bacterial cell growth and protection. Results: The growth and cell wall integrity of the modified Klebsiella pneumoniae (UPM ESBLKP1) were significantly inhibited and reduced, respectively. Interestingly, wild type Klebsiella pneumoniae showed a normal growth curve while modified strains showed a faster doubling rate when supplemented with Luria-Bertani media. In contrast, slower growth rate of modified strain was observed in the M9 minimal media. This explained the higher doubling rate of mutants on nutrient rich medium earlier is being related to gene recovery. They grew slowly in the minimal media as they were adapting to a new environment while recovering the uppP gene and surviving, proving the success of its gene modification. Conclusion: The developed CRISPR-gRNA system was able to modify the targeted Klebsiella pneumoniae gene hence providing an opportunity to develop a new drug for Klebsiella pneumoniae infection as an alternative to antibiotics.
Fulltext Vaccines and vaccination against infectious bursal disease of chickens: prospects and challenges

Liew, Pit Sze, Nurulfiza Mat Isa, Omar Abdul Rahman, Aini Ideris, Mohd HAIRBEJO

The Pertanika Journal of Scholarly Research Reviews, 2016;2(2):23-39.
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Infectious bursal disease (IBD), also known as the Gumboro disease, has been a great
concern for poultry industry worldwide. The first outbreak of IBD due to very virulent (vv) IBD virus
(IBDV) infection in Malaysia was reported in 1991. The major economic impact of the disease is high
mortality and poor performance. The virus causes immunosuppression where if the infected chicken
recovered from the acute disease, they become more susceptible to infections of other pathogens and
fail to respond to vaccines. Therefore, prevention is important and vaccination has become the
principal control measure of IBDV infection in chickens. The conventional attenuated live and killed
vaccines are the most commonly used vaccines. With the advancement of knowledge and technology,
new generation of genetically-engineered vaccines like viral vector and immune complex vaccines
have been commercialised. Moreover, hatchery vaccination is becoming a common practise, in
addition to farm vaccination. Currently, the disease is considerably under controlled with the
introduction of vaccination. However, occasional field outbreaks are still commonly reported. The
demand for vaccines that could suit the field situation continues to exist. The endemicity of disease,
presence of challenge in the farm and maternally derived antibody in chicks are affecting the choice
vaccine as well as the vaccine development and vaccination strategies. In this review, advances made
in various vaccines that have been commercialised or under development, and challenges that they
face, are outlined. Furthermore, how the emergence of vvIBDV affect the progress of vaccine
development and influence its vaccination strategy are discussed.
Fulltext Efficacy, humoral, and cell-mediated immune response of inactivated fowl adenovirus 8b propagated in chicken embryo liver cells using bioreactor in broiler chickens

Ugwu CC, Hair-Bejo M, Nurulfiza MI, Omar AR, Ideris A

Vet World, 2022 Nov;15(11):2681-2692.
PMID: 36590109 DOI: 10.14202/vetworld.2022.2681-2692

BACKGROUND AND AIM: Fowl adenovirus (FAdV) 8b causes inclusion body hepatitis, resulting in major economic losses globally among chickens. The objectives were to inactivate FAdV 8b isolate propagated in chicken embryo liver (CEL) cells using a stirred tank bioreactor (UPM08136P5B1) and determine the humoral and cell-mediated immune response, efficacy, and virus shedding in broiler chickens.
MATERIALS AND METHODS: The FAdV 8b isolate UPM08136P5B1 was inactivated using binary ethyleneimine, adjuvanted with Montanide 71VG, inoculated into day-old broiler chickens in a booster group (BG) and non-booster group (NBG), and challenged with a pathogenic FAdV 8b strain. Clinical signs, gross lesions, body weight (BW), liver: body weight ratio, FAdV antibody titer using enzyme-linked immunosorbent assay, and histopathological changes were recorded. The CD3+, CD4+, and CD8+ T-lymphocyte profiles of the liver, spleen, and thymus using flow cytometry, and viral load in liver and cloacal shedding using quantitative polymerase chain reaction were evaluated.
RESULTS: Chickens in the challenged control group (CCG) exhibited mild clinical signs, gross lesions, and histopathological changes, which were absent in the inoculated groups, and had lower BW and higher liver BW ratio than chickens in the unchallenged control group (UCG); BG and NBG on 35- and 42-days post-inoculation (DPI). Chickens in NBG and BG had higher antibodies than UCG on 7, 21, 35, and 42 DPI. The challenged BG and NBG produced higher antibodies than the CCG on 35 DPI. T-lymphocytes were higher among the inoculated groups than UCG in the liver, spleen, and thymus. Inoculated challenged groups recorded higher CD3+, CD4+, and CD8+ T-lymphocytes on 35 and 42 DPI than CCG. The challenged control group had a significantly higher viral load in the liver than challenged that in BG on 35 DPI and BG and NBG on 42 DPI. The challenged control group had significantly higher challenge FAdV shedding than challenged inoculated groups on 35 and NBG on 42 DPI.
CONCLUSION: UPM08136P5B1 was successfully inactivated and mixed with Montanide 71VG. The inactivated vaccine candidate that induced humoral and cellular immunity was effective, reduced FAdV load in the liver, and shedding in the cloaca, and could be useful against FAdV 8b infections in chickens.