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Porcine DNA authentication in commercial products is critical due to improper or non-certified Halal logo on its packaging. This study detected the presence of porcine DNA in particularly processed meat products sold in Sarawak. A total of 75 samples were collected, consisting of poultry, beef, and seafood products. DNA was isolated and amplified in a polymerase chain reaction that targeted cytochrome B (cytb) and mitochondrial D-loop. PCR products were analysed via gel electrophoresis and viewed through gel documentation. The positive result was observed in 5 samples: 3 from poultry and 2 from beef. The DNA band were detected at an amplicon size of 174 base pairs for poultry products whereas 100 base pairs for beef products. Porcine DNA was absent in seafood products. Positive samples were validated through DNA sequencing. Nucleotide sequences from DNA sequencing were compared with the database using the Basic Local Alignment Search Tool (BLAST). BLAST analysis indicated percentages ranging from 96.83% to 100%, demonstrating high similarity with porcine DNA; one of the samples was labelled with a foreign Halal logo. In summary, this study provides the groundwork for further inspection of the Halal status in commercially processed meat products available in Sarawak.

This case report describes a 49-year-old immunocompromised woman with tenosynovitis of the left middle finger caused by Group B Streptococcus (GBS). She claimed that a fishbone picked over her left middle finger. An orthopaedic surgeon operated for incision and drainage of pus discharge, wound debridement of the left middle finger and A1 and A2 pulley release. Treatment was initiated with parenteral cefepime three times per day given the growth of mixed Enterobacter species on the culture media and continued with oral cefuroxime twice daily upon discharge for one week. Unfortunately, during the orthopaedic clinic follow-up, the wound was unclean with a slough and skin necrotic patch. Therefore, Ray’s amputation of the left middle finger proceeded. This case contributes to further investigation of the GBS tenosynovitis due to the rise in GBS invasive infections and shows the importance of early diagnosing and initiating treatment with antibiotics that are effective against this pathogen.

To explore Tuberculosis (TB) health promotion and education in universities, and to provide a feasible reference method and specific implementation measures for improving health education among university students. We collected a total of 811 respondents, using a uniform questionnaire online survey questionnaire network. The completed questionnaires were double-entered using EpiData3.1, the database was established, and the analysis was performed by Excel2016 and SPSS 22 software. The total awareness rate of the 8 core information on tuberculosis prevention and control was 74.2 %. Among them, “the state provides free anti-tuberculosis drugs and major tests for infectious tuberculosis patients” with the lowest awareness rate of 49.2 %; followed by the awareness rate of “should care about tuberculosis patients and should not discriminate against tuberculosis patients”, for 63.8 %. The channels for college students to acquire knowledge about tuberculosis prevention were “newspapers and magazines”, accounted for 50.3 percent; “broadcasting, television and video” accounted for 52.4 %; “wall advertising, bulletin boards, slogans” accounted for 44.6 %; “School Health Education” accounted for 38.5 % of “school propaganda columns or publicity panels” accounted for 34.9 %. The favourite way for college students to promote was “watching TV” 40.3 %.

Suhaili Shamsi,1 Ahmad Ashraful Hadi Abdul Ghafor,2 Nur Hazwani Norjoshukrudin,1 Ida May Jen Ng,1 Siti Nur Sharmila Abdullah,1 Seri Narti Edayu Sarchio,2 Faizah Md Yasin,3,4 Shafinaz Abd Gani,1 Mohd Nasir Mohd Desa2 1Laboratory of Animal Biochemistry and Biotechnology, Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia; 2Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia; 3Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia; 4Institute of Nanoscience and Nanotechnology (ION2), Universiti Putra Malaysia, Serdang, Selangor, 43400, MalaysiaCorrespondence: Suhaili Shamsi, Laboratory of Animal Biochemistry and Biotechnology, Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia, Tel +603-9769 7964, Fax +603-9769 7590, Email sh_suhaili@upm.edu.myBackground: The impetuous usage of antibiotics has led to the perpetual rise of methicillin-resistant Staphylococcus aureus (MRSA), which has garnered the interest of potential drug alternatives, including nanomaterials.Purpose: The present study investigates the stability, toxicity, and antibacterial potential of gallic acid-loaded graphene oxide (GAGO) on several MRSA strains.Methods: The stability of a synthesized and characterized GAGO was monitored in different physiological media. The toxicity profile of GAGO was evaluated in 3T3 murine fibroblast cells and the embryonic zebrafish model. The antibacterial activity of GAGO against MRSA, methicillin-susceptible S. aureus (MSSA), and community-acquired MRSA; with or without Panton-valentine leucocidin gene (MRSA-pvl+ and MRSA-pvl-) was investigated through disk diffusion, CFU counting method, time-kill experiment, and high-resolution transmission electron microscopy (HRTEM) observation.Results: A stable GAGO nanocomposite has shown an improved toxicity profile in 3T3 murine fibroblast cells and zebrafish embryos, besides exhibiting normal ROS levels than graphene oxide (GO) and GA (gallic acid). The nanocomposite inhibited the growth of all bacterial strains employed. The effectiveness of the GAGO nanocomposite was comparable to cefoxitin (CFX), at ≥ 150 μg/mL in MRSA and MSSA. GAGO exhibited a significantly delayed response towards MRSA-pvl+ and MRSA-pvl-, with increased inhibition following 8 to 24 h of exposure, while comparable activity to native GA was only achieved at 24 h. Meanwhile, for MRSA and MSSA, GAGO had a comparable activity with native GA and GO as early as 2 h of exposure. HRTEM observation further reveals that GAGO-exposed cells were membrane compromised.Conclusion: In summary, the present study indicates the antibacterial potential of GAGO against MRSA strains, but further study is warranted to understand the mechanism of action of GAGO and its resistance in MRSA strains.Keywords: graphene oxide, gallic acid, nanomaterial, MRSA, toxicity, multidrug resistance

This study authenticated fish feed sources and determined lard adulteration using dataset pre-processing, principal component analysis (PCA), discriminant analysis (DA) and partial least square regression (PLSR) on 19 triacylglycerols (TAGs) and 16 thermal properties (TPs). At cumulative variability (90.625%) and Keiser-Meyer Olkin (KMO) value (0.811), the PCA identified 10 TAGs and 3 TPs with strong factor loading. The dioleoyl-1-palmitoyl glycerol (POO), dipalmitoyl-3-oleoyl glycerol (PPO) and dipalmitoyl-1-linoleoyl (PPL) characterized fish feeds containing palm oil while dilinoleoyl-1-oleoyl glycerol (OLL), dilinoleoyl-1-palmitoyl (PLL), dioleoyl-3-linoleoyl glycerol (OOL), initial cooling temperature (ICT), palmitoyl-oleoyl-linoleoyl glycerol (POL), palmitoyl-stearoyl-oleoyl glycerol (PSO) and final heating temperature (FHT) characterized lard-containing fish feeds. The DA had successfully classified the fish feed sources and selected the PPL, POL, PPO, OOL, ICT, PLL, FHT, POO and OLL as the most influential biomarkers for the authentication purpose. The Z-test result (p>0.05) indicated that the PLSR could determine the percentage of lard adulteration in fish feed. Hence, incorporating multivariate and instrumental analyses could authenticate the fish feed sources.

Pilus has been recently associated with pneumococcal pathogenesis in humans. The information regarding piliated isolates in Malaysia is scarce, especially in the less developed states on the east coast of Peninsular Malaysia. Therefore, we studied the characteristics of pneumococci, including the piliated isolates, in relation to antimicrobial susceptibility, serotypes, and genotypes at a major tertiary hospital on the east coast of Peninsular Malaysia. A total of 100 clinical isolates collected between September 2017 and December 2019 were subjected to serotyping, antimicrobial susceptibility test, and detection of pneumococcal virulence and pilus genes. Multilocus sequence typing (MLST) and phylogenetic analysis were performed only for piliated strains. The most frequent serotypes were 14 (17%), 6A/B (16%), 23F (12%), 19A (11%), and 19F (11%). The majority of isolates were resistant to erythromycin (42%), tetracycline (37%), and trimethoprim-sulfamethoxazole (24%). Piliated isolates occurred in a proportion of 19%; 47.3% of them were multidrug-resistant (MDR) and a majority had serotype 19F. This study showed ST236 was the most predominant sequence type (ST) among piliated isolates, which was related to PMEN clone Taiwan19F-14 (CC271). In the phylogenetic analysis, the piliated isolates were grouped into three major clades supported with 100% bootstrap values. Most piliated isolates belonged to internationally disseminated clones of S. pneumoniae, but pneumococcal conjugate vaccines (PCVs) have the potential to control them.

Low DNA concentration recovered from highly processed products such as gelatin and gelatin-based products renders difficulty in detecting porcine contamination using conventional PCR techniques. We documented here a porcine-specific loop-mediated isothermal amplification (LAMP) to identify porcine traces in gelatin products. The porcine-specific primers were designed according to mitochondrial DNA of Cytochrome b gene sequence. Here we used two different reaction mixtures for LAMP assay (GENIE and MYRM) against the same DNA samples extracted from gelatin products and porcine-specific primers to detect the presence of porcine DNA. The porcine-specific primers were shown to be specific only to Sus scrofa against 14 DNA of other meat species. The analytical sensitivity of the LAMP assay for porcine DNA detection is 1 pg/µL using both GENIE (within 30 m) and MYRM (within 60 m) reaction mixtures. Analysis against 32 samples of gelatin products showed that five samples were found to contain porcine DNA; two samples out of six gelatin powder samples and three gelatin capsule samples out of nine. Out of these five positive samples, three were not labeled containing porcine gelatin. Overall, LAMP assay in this study showed an excellent specificity, sensitivity and rapidity in detection of porcine DNA in gelatin products. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-020-04932-2).

This study investigated antibacterial activities, chemical composition and extract efficacy of Carica papaya peel (CPPE) var. Sekaki/ Hong Kong. Nine green solvents were used to extract the Carica papaya peel, and the extracts were subjected to antibacterial tests and assays against 14 bacteria. The most potent extract was then subjected to phenolic and flavonoid assays, gas chromatography-mass spectrometry (GC/MS) analysis, and efficacy study on food model systems. All CPPEs showed antibacterial activities, and pentane extract had moderate to clear inhibitions against all 14 bacteria. Ethanol extract of Carica papaya peel (ECPPE) inhibited C. perfringens, L. monocytogenes, B. subtilis, V. parahaemolyticus, and V. vulnificus with a minimum inhibitory concentration (MIC) of 1.563 mg/ml; therefore, the ECPPE was selected as the most potent extract. The total phenolic (TPC) and flavonoid contents (TFC) of the CPPEs ranged between 6.20 to 58.75 mg GAE/g DW and 1.35–29.09 mg QE/g DW, respectively. Palmitic acid, linoleic acid, β-sitosterol and stigmasterol in ECPPE may be potential antibacterial compounds that render antibacterial activities. The ECPPE could effectively inhibit the tested bacteria in low carbohydrate and high protein, fat, and fibre food model systems, which may suit the investigation of ECPPE efficacy in meat, poultry and dairy products, high fat-content products such as mayonnaise, butter and margarine and high fibre food such as legumes, lettuce and barley.

The aim of this study was to study the genotype of a hospital collection of Group B Streptococcus (GBS) from invasive and non-invasive sites. Fifty-one pre-characterised human of GBS were re-identified and further analysed by multilocus sequence typing (MLST) in relation to previously published serotypes. Fifteen sequence types (ST) were found with ST1 being the most predominant. ST1 was also associated with majority of the invasive isolates. The genotypic distribution patterns of GBS in this study were largely in agreement with previous reports from other countries indicating the tendency of certain genotypes to prevail in human infection settings.

Allah has instructed all humankind to eat permissible healthy food according to His guidance as narrated in Al-Quran. Following Maqasid as-Shari’ah, it is a Darruriyyah to ensure the safety of food as an act of protecting life. Therefore, the application of antibacterial agent on food aims to improve the consumers’ health and control the herd immunity. Plant seed is a newly emerged alternative as a source of antibacterial agents. Commonly discarded, the utilisation of plant seed will not only reduce environmental waste issue but also benefited economically to many industries. This article was aimed to review the current literature on antibacterial test and toxicity of various plant seed extracts that potentially contributed to the efficacy of the seed. This review presents a comprehensive analysis of the antibacterial activity assay of the plants' seeds. It covers endpoint and descriptive methods, plant seed toxicity and application of the seed extract in food industries. This review was conducted using the various science-based platform such as Science Direct, Google Scholar, Emerald etc. The keywords ‘antibacterial’, ‘antibacterial seed’, ‘seed extract’ and ‘toxicity seed’ was used in this search.

This study authenticated fish feed sources and determined lard adulteration using dataset pre-processing, principal component analysis (PCA), discriminant analysis (DA) and partial least square regression (PLSR) on 19 triacylglycerols (TAGs) and 16 thermal properties (TPs). At cumulative variability (90.625%) and Keiser-Meyer Olkin (KMO) value (0.811), the PCA identified strong factor loading variables, i.e., OLL, PLL, OOL, POL, PPL, POO, PPO, PSO, ICT and FHT in PC1 and LLLn, OOO and CT2 in PC2. These variables were significantly (p < 0.05) contributing to lard-palm-oil (L-PO) clusters: (1) POO, PPO and PPL (high loading) and OLL, PLL, OOL, ICT, POL, PSO and FHT (low loading) in 0:100 and 25:75 L-PO clusters; (2) CT2, OOO and LLLn (high loading) in 50:50 L-PO cluster; and (3) OLL, PLL, OOL, ICT, POL, PSO and FHT (high loading) and POO, PPO and PPL (low loading) in 72:25 and 100:0 L-PO clusters. Training, validation and testing datasets had 100%, 84.44% and 100% correct-classification, respectively at p < 0.0001 of Wilks' lambda and p < 0.0001 Fisher distance. The DA selected PLL, OOL, POL, PPL, PSO, ICT and FHT as the significantly authenticating biomarkers (p < 0.05). With determination coefficient (R²) (0.9693), mean square error (MSE) (38.382) and root mean square error (RMSE) (6.195), the PLSR's variable importance in the projection (VIP) identified the most influential biomarkers, i.e., PPL, POL, PPO, OOL, ICT, PLL, FHT, POO and OLL. The Z-test result (p > 0.05) indicated that the PLSR could determine the lard adulteration percentage in fish feed.

Background Pneumococcal pneumonia is the leading cause of under-five mortality globally. The surveillance of pneumococcal serotypes is therefore vital for informing pneumococcal vaccination policy and programmes. Pneumococcal conjugate vaccines (PCVs) have been available as an option in the private healthcare setting and beginning December 2020, PCV10 was incorporated as part of routine national immunisation programme (NIP) in Malaysia. We searched existing literature on pneumococcal serotype distribution across Malaysia to provide an overall view of this distribution before the implementation of PCV10. Methods Online databases (PubMed, Ovid MEDLINE and Scopus), reference lists of articles identified, and grey literature (Malaysian Ministry of Health website, WHO website) were systematically searched for relevant literature on pneumococcal serotype distribution across Malaysia up to 10th November 2020. No lower date limit was set to maximise the number of target reports returned. Results of serotypes were split by age categories, including ≤5 years, > 5 years and unreported for those that did not specify. Results The search returned 18 relevant results, with a total of 2040 isolates. The most common serotypes across all disease types were 19F (n = 313, 15.3% [95%CI: 13.8–17.0]), 23F (n = 166, 8.1% [95%CI: 7.0–9.4]), 14 (n = 166, 8.1% [95%CI: 7.0–9.4]), 6B (n = 163, 8.0% [95%CI: 6.9–9.2]) and 19A (n = 138, 6.8% [95%CI: 5.8–7.9]). Conclusion Four of the most common serotypes across all isolate sources in Malaysia are covered by PCV10, while PCV13 provides greater serotype coverage in comparison to PCV10. There is still a need for surveillance studies, particularly those investigating serotypes in children under 5 years of age, to monitor vaccine effectiveness and pneumococcal population dynamic following implementation of PCV10 into routine immunisation.

Background: Streptococcus pyogenes has a variety of virulence factors and the predominant invasive strains differ according to specific emm types and geographical orientation. Although emm typing is commonly used as the gold standard method for the molecular characterization, multilocus sequence typing (MLST) has become an important tool for comparing the genetic profiles globally. This study aimed to screen selected virulence genes from invasive and non-invasive clinical samples and to characterize the molecular epidemiology by emm typing and MLST methods. Methods: A total of 42 S. pyogenes isolates from invasive and non-invasive samples collected from 2014 to 2015 from two different tertiary hospitals were investigated for the distribution of virulence factors and their molecular epidemiology by emm and multilocus sequence typing methods. Detection of five virulence genes (speA , speB , speJ , ssa and sdaB) was performed using multiplex polymerase chain reaction (PCR) using the standard primers and established protocol. Results: Multiplex PCR analysis revealed that sdaB/speF (78.6%) and speB (61.9%) were the predominant virulence genes. Regardless of the type of invasiveness, diverse distribution of emm types/subtypes was noted which comprised of 27 different emm types/subtypes. The predominant emm types/subtypes were emm63 and emm18 with each gene accounted for 11.8% whereas 12% for each gene was noted for emm28, emm97.4 and emm91. The MLST revealed that the main sequence type (ST) in invasive samples was ST402 (17.7%) while ST473 and ST318 (12% for each ST) were the major types in non-invasive samples. Out of 18 virulotypes, Virulotype A (five genes, 55.6%) and Virulotype B (two genes, 27.8%) were the major virulotypes found in this study. Phylogenetic analysis indicated the presence of seven different clusters of S. pyogenes. Interestingly, Cluster VI showed that selected emm/ST types such as emm71/ST318 (n=2), emm70.1/ST318 (n=1), emm44/ST31 (n=1) and emm18/ST442 (n=1) have clustered within a common group (Virulotype A) for both hospitals studied. Conclusion: The present study showed that group A streptococci (GAS) are genetically diverse and possess virulence genes regardless of their invasiveness. Majority of the GAS exhibited no restricted pattern of virulotypes except for a few distinct clusters. Therefore, it can be concluded that virulotyping is partially useful for characterizing a heterogeneous population of GAS in hospitals.

Analysis of halal food requires that the laboratories conductive the tests adhere to international guidelines and standards. Common worldwide guidelines and standards for laboratory include the ‘International Organization for Standardization 17025’ (ISO 17025), the ‘Good Manufacturing Practice’ (GMP), and ‘Good Laboratory Practice’ (GLP). In halal analysis, the laboratory shall comply with ISO 17025, GMP, and ‘Good Hygiene Practice’ (GHP) as stated in the ‘Manual Procedure for Malaysia Halal Certification’ (MPPHM). This article discusses the basic requirements for laboratory practises, specifically for halal analysis. The study compares these international guidelines with ‘Malaysia Halal Standards’ to demonstrate that these international standards are combined with Islamic practices to produce valid test results using globally recognized best-practices. This promotes confidence in the halal laboratory’s work both nationally and internationally, and will thus improve international trade. Keywords: Halal analysis, ISO 17025, GMP, GLP, laboratory operation.

Highlights • A short-amplicon based (69 bp) SP-PCR assay was used for feline meat detection. • No cross-species amplification from total twenty four (24) other species of land, aquatic and plant origins. • The 69 bp target was stable under autoclaving and boiling condition of the commercial products. • The visualized RFLP profiles were clear and stable. And the assay was proved to be feasible in screening commercial products. • Screening result of total 378 sample across Malaysia were negative for feline meat detection, Determination of feline meat in food products is an important issue for social, health, economic and religious concern. Hence this paper documented the application of species specific polymerase chain reaction-restriction fragment length polymorphism (SP-PCR-RFLP) assay targeting a short-fragments (69 bp) of mitochondrial cytochrome b (cytb) gene to screen feline meat in commercial meat products using lab-on-a-chip. The SP-PCR assay proved its specificity theoretically and experimentally while testing with different common animal, aquatic and plant species of DNA. The feline specific (69 bp, 43- and 26-bp) characteristic molecular DNA pattern was observed by SP-PCR and RFLP analysis. For assay performance, it was tested in three different types of commercial dummy meat products such as frankfurters, nuggets and meatballs and digested with AluI-restriction enzyme. The highest sensitivity of the assay using lab-on-a-chip was as low as 0.1 pg or 0.01% (w/w) in commercial dummy meat products. We have also applied this assay to screen three important commercial meat products of six different brand from six supermarket chains located at three different states of Malaysia. Thus total 378 samples were tested to validate the specificity, sensitivity, stability of the assay and utilization of it for commercial meat product screening.

High-quality DNA extracts are imperative for downstream applications in molecular identification. Most processed food products undergo heat treatments causing DNA degradation, which hampers application of DNA-based techniques for food authentication. Moreover, the presence of inhibitors in processed food products is also problematic, as inhibitors can impede the process of obtaining high qualities and quantities of DNA. Various approaches in DNA extraction and factors in structure and texture of various food matrices affecting DNA extraction are explained in this review.

This study attempted to identify and determine distribution of Leptospira spp. in environmental samples using 16S rRNA and rpoβ genes amplification. The samples were collected from high risk areas in Selangor, Malaysia. A total of 105 environmental samples consisting of soil and water were subjected to direct DNA extraction and PCR reaction. PCR products were analysed using gel electrophoresis and subjected to sequence analysis. Thirteen out of 105 (12.38%) samples were amplified for 16S rRNA with an expected amplicon size of 330 bp, while 50 out of 105 (47.62%) samples showed amplification using rpoβ primers, but were not of expected size. Of the 13 16S rRNA amplified samples, only 5 were identified as Leptospira in the gene sequence analysis and clustered under uncultured group via phylogenetic tree. This study showed the DNA-based approach using PCR and sequence analysis is able to detect the presence of Leptospira, although environmental samples may contain diverse microbial populations that may complicate the detection. Overall, the study suggested the importance of surveillance for Leptospira from environmental samples.

This review aims to elucidate the different mechanisms of blood brain barrier (BBB) disruption that may occur due to invasion by different types of bacteria, as well as to show the bacteria-host interactions that assist the bacterial pathogen in invading the brain. For example, platelet-activating factor receptor (PAFR) is responsible for brain invasion during the adhesion of pneumococci to brain endothelial cells, which might lead to brain invasion. Additionally, the major adhesin of the pneumococcal pilus-1, RrgA is able to bind the BBB endothelial receptors: polymeric immunoglobulin receptor (pIgR) and platelet endothelial cell adhesion molecule (PECAM-1), thus leading to invasion of the brain. Moreover, Streptococcus pneumoniae choline binding protein A (CbpA) targets the common carboxy-terminal domain of the laminin receptor (LR) establishing initial contact with brain endothelium that might result in BBB invasion. Furthermore, BBB disruption may occur by S. pneumoniae penetration through increasing in pro-inflammatory markers and endothelial permeability. In contrast, adhesion, invasion, and translocation through or between endothelial cells can be done by S. pneumoniae without any disruption to the vascular endothelium, upon BBB penetration. Internalins (InlA and InlB) of Listeria monocytogenes interact with its cellular receptors E-cadherin and mesenchymal-epithelial transition (MET) to facilitate invading the brain. L. monocytogenes species activate NF-κB in endothelial cells, encouraging the expression of P- and E-selectin, intercellular adhesion molecule 1 (ICAM-1), and Vascular cell adhesion protein 1 (VCAM-1), as well as IL-6 and IL-8 and monocyte chemoattractant protein-1 (MCP-1), all these markers assist in BBB disruption. Bacillus anthracis species interrupt both adherens junctions (AJs) and tight junctions (TJs), leading to BBB disruption. Brain microvascular endothelial cells (BMECs) permeability and BBB disruption are induced via interendothelial junction proteins reduction as well as up-regulation of IL-1α, IL-1β, IL-6, TNF-α, MCP-1, macrophage inflammatory proteins-1 alpha (MIP1α) markers in Staphylococcus aureus species. Streptococcus agalactiae or Group B Streptococcus toxins (GBS) enhance IL-8 and ICAM-1 as well as nitric oxide (NO) production from endothelial cells via the expression of inducible nitric oxide synthase (iNOS) enhancement, resulting in BBB disruption. While Gram-negative bacteria, Haemophilus influenza OmpP2 is able to target the common carboxy-terminal domain of LR to start initial interaction with brain endothelium, then invade the brain. H. influenza type b (HiB), can induce BBB permeability through TJ disruption. LR and PAFR binding sites have been recognized as common routes of CNS entrance by Neisseria meningitidis. N. meningitidis species also initiate binding to BMECs and induces AJs deformation, as well as inducing specific cleavage of the TJ component occludin through the release of host MMP-8. Escherichia coli bind to BMECs through LR, resulting in IL-6 and IL-8 release and iNOS production, as well as resulting in disassembly of TJs between endothelial cells, facilitating BBB disruption. Therefore, obtaining knowledge of BBB disruption by different types of bacterial species will provide a picture of how the bacteria enter the central nervous system (CNS) which might support the discovery of therapeutic strategies for each bacteria to control and manage infection.

Objective The high prevalence of leptospirosis in humans is of great public health concern, particularly in tropical and subtropical regions. This study aimed to determine the seroprevalence of leptospiral antibodies and distribution of serovars, and to assess the usefulness of enzyme-linked immunosorbent assay (ELISA) as a screening method for leptospiral antibodies in a high-risk healthy community. Methods Cross-sectional study of 231 market workers and food handlers in wet markets and food premises from two localities in central Malaysia. Respondents' background information was obtained using a questionnaire. Serum samples were tested for leptospiral antibodies using ELISA and microscopic agglutination test (MAT). Results Seroprevalence of leptospirosis among healthy workers was 46.3%. Detection of seropositivity was higher by MAT (46%) than ELISA (15%). We observed high seropositivity among local workers (49%), food handlers (49.5%), females (60.8%) and those aged 34 years and older (46.3%). Local strain LEP175 was the predominant serovar, followed by WHO strain Patoc. Conclusion Overall seroprevalence among healthy food handlers and market workers was high in this study. The workplace places susceptible individuals at risk of leptospirosis.

The oils obtained from three Indonesian avocado (Persea americana) cultivars namely Merah bundar, Ijo bundar and Ijo panjang, were compared to the oils obtained from imported Fuerte and Shepard avocado varieties in terms of lipid characteristics and tocopherol content. The oils of all avocado varieties existed in a semisolid form, except that of the Fuerte variety. Free fatty acids and peroxide levels in the oils obtained from locally grown avocado were lower than the levels in the oils obtained from imported avocados, implying that local avocado oils have a better oxidative stability. Interestingly, the tocopherol (alpha, beta, gamma and delta) content of local avocado oils was also much higher than that of imported avocado oils. Meanwhile, differences in the FTIR spectrum of avocado oils were found at frequencies of 1034 and 968 cm−1. There were also some differences in the fatty acid and triacylglycerol composition of avocado oils. Due to these differences, the physicochemical characteristics and the solidification and thermal profiles of the oils obtained from local avocado cultivars were completely different from those of the imported avocado varieties (Fuerte and Shepard).

Beef, buffalo and pork are the major meat of economic, religious and health concern. Current methods to authenticate these materials in food chain are based on mainly single gene targets which are susceptible to break down by food processing treatments. We, for the first time, described here a double gene targeting short-amplicon length multiplex polymerase chain reaction assay for discriminating bovine, buffalo and porcine materials in a single assay platform. The advantage of the assay is evidenced in terms of fidelity, cost and time since it is highly unlikely that two different targets would be missing even in a decomposed specimen. Detection of multiple targets in a single assay definitely saves analytical cost and time. Mitochondrial cytochrome b (cytb) and ND5 genes were targeted and six different targets (length: 90–146 bp), two for each of cow (120 and 106bp), buffalo (90 and 138bp) and pig (73 and 146bp), were amplified from raw, boiled, autoclaved and microwaved cooked meat under pure and mixed matrices. The detection limit was 0.02 ng DNA under pure states and 0.1% meat in binary mixtures and meatball products. Screening of Malaysian meatball products revealed all beef products were buffalo positive in which 35% were totally replaced. In contrast, all pork products were found uncontaminated from beef and buffalo.

Summary: Background/purpose: The purpose of this study is to characterize GBS isolates that were collected from three major hospitals in a densely populated area of Klang Valley for their demographics, serotypes, antibiotic susceptibility patterns and genetic background. Methods: Sixty GBS isolates from sterile and non-sterile samples in three major hospitals in the Klang Valley area of Malaysia were collected by convenience sampling from 2012 until March 2014. These isolates were studied for their antimicrobial susceptibilities, serotypes and genotypes. Patients’ demographic data and clinical information were collected from lab request forms. Results: Diabetes mellitus was the only underlying condition (7 patients, 23.3%); the remaining samples were from patients who were immunocompromised due to medications. Fifty-nine (98%) isolates were sensitive to penicillin, while 78.3% and 88.3% of the isolates were sensitive to erythromycin and clindamycin, respectively. Serotype Ia was the most common serotype (n = 27, 45%), followed by serotype III (n = 10, 16.7%), V (n = 9, 15%), VI (n = 8, 13.3%), VIII (n = 2, 3.3%) and VII (n = 1, 1.7%). Random Amplified Polymorphic DNA (RAPD) typing showed a diverse genetic pedigree for all isolates, including four major groups that clustered according to geographical location. Conclusion: This preliminary study determines the prevalence of limited common serotypes and antimicrobial resistance in distinct GBS isolates. Nonetheless, the RAPD clustering pattern suggests a close genetic lineage of the GBS isolates based on their isolation sites and location of hospitals. Keywords: Group B Streptococcus, Resistant, Serotype

Fish oil is a common source of fat in fish feed production. However, there is a tendency to substitute fish oil with other fats such as lard to reduce production costs. Thus, an efficient method for lard detection is highly needed for fish feed’s authenticity. In this study, sn-2 fatty acids (sn-2 FAs) and fatty acid (FA) compositions were incorporated with chemometric techniques namely Principal Component Analysis (PCA), Orthogonal Partial Least Square-Discriminant Analysis (OPLS-DA), and Orthogonal Partial Least Square-Regression (OPLS-R) to identify lard adulteration in the fish feeds. The inclusion of sn-2 FAs into PCA model 2 exhibited a preferable variation pattern relative to PCA model 1. The PCA identified C14:0, C18:0, C18:2, C18:3, C20:0 sn-2 C16:0, sn-2 C18:0, sn-2 C18:1, and sn-2 C18:2 were the most significant FAs to discriminate the fish feeds. The inclusion of sn-2 FA composition improved the OPLS-DA model 2 performance by providing more significant class discrimination between lard-adulterated, and non-adulterated fish feeds as compared to OPLS-DA model 1. The OPLS-DA model 2 identified C18:0, C18:2, C18:3, and sn-2 C16:0 FAs as markers of lard adulteration with an increment in the value of the coefficient of determination (R2) and decrement in the Root Mean Square Error of Estimation (RMSEE) and Root Mean Standard of Cross-Validation (RMSECV) values. The Support Vector Machine (SVM), Random Forest (RF), Multilayer Perceptron-Artificial Neural Network (MLP-ANN), and internal and external validations corroborated the OPLS-DA model 2 and OPLS-R model 2 performances. Therefore, the incorporation of sn-2 FA and FA compositions coupled with the chemometric techniques had improved the detection and quantification of lard adulteration in fish feeds.

The present study was carried out to compare the antimicrobial activities of methanol leaf extracts of Bauhinia purpurea, Dicranopterislinearis, Melastomamalabathricum and Muntingiacalabura portrayed by different antimicrobial assays against methicillin-susceptible and methicillin- resistant Staphylococcus aureus strains. Antimicrobial activities of the methanol leaf extracts were preliminarily screened by disc diffusion. Minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth microdilution and colorimetric assay (resazurin). Based on disc diffusion method, S. aureusATCC®700699™ (MRSA) elucidated higher susceptibility pattern against all plant extracts compared to S. aureusATCC®25923™ (MSSA). Taking results from all employed assays into consideration, M. calaburamethanol leaf extract comparably elicited the highest antimicrobial activity than the other methanol leaf extracts against both microorganisms. The MIC values were determined by colorimetric assay (resazurin) due to pigmentations of the methanol leaf extracts that obscured visual growth turbidity inspection. Complication in colour changes observation in colorimetric assay to determine MBC was overcome by employing the conventional plating method. This study suggested that all antimicrobial assays should be carried out concurrently so as the data obtained can be comparatively analysed for a better outcome as each antimicrobial assay has its own shortfall.

Pneumococci are a common cause of severe infections, such as otitis media, pneumonia, meningitis and bacteremia. Pili are detected in a small proportion of pneumococcal population, but these structures have recently been associated with bacterial virulence in humans. Therefore, the epidemiological relationships between pneumococcal pili, serotype and antimicrobial resistance are of interest. This study aims to discuss the virulence contribution of the Streptococcus pneumoniae pili and the epidemiological relationships among the pilus genes, antimicrobial resistance trends, regional serotypes and genotypic variations. Previous reports have characterized the pneumococcal pilus islet as a clonal feature in the pneumococcal serotypes that are covered by the pneumococcal conjugate vaccine (PCV), including serotypes 19A, 19F, 23F and 7F. Many of the pneumococcal molecular epidemiology network (PMEN) clones are piliated isolates that are also strongly associated with a high frequency of multidrug resistance. Most of these piliated pneumococcal isolates belong to a few clonal complexes (CC), such as CC320, CC199, CC271, CC191 and CC156. Additional molecular epidemiology and genomic studies, particularly whole genome sequence analysis (WGS), are needed to develop an in-depth understanding of the piliated pneumococcal isolates.

The present study focuses on the antimicrobial activity and safety aspect of lactobacilli strains isolated from the anogenital region. Sixty-two lactobacilli strains were isolated from 52 healthy, non-menstruating women from a local health clinic in Puchong, Selangor, Malaysia and confirmed by 16S rDNA sequencing. The samples were collected by sterile cotton swabs and directly streaked onto De Man, Rogosa, Sharpe agar prior to transportation to the lab on ice. A selective preliminary screening against several Candida albicans and Candida glabrata strains identified 5 promising lactobacilli strains for the subsequent assays; L. delbreuckii 45E, L. fermentum 28E, L. mucosae 28C, L. reuteri 29A and L. reuteri 29B. A follow-up antibacterial assay identified L. delbreuckii 45E, L. reuteri 29A and L. reuteri 29B as the lactobacilli strains with the highest inhibitory activity. Subsequent assays including haemolytic activity, susceptibility to antibiotics, quantification of D/ l -lactic acid and H2O2 production as well as bile salt hydrolase (BSH activity) indicated that L. reuteri 29B was a promising probiotic candidate. An acute toxicity study in vivo involving the administration of 1 × 108, 1 × 109 and 1 × 1010 cfu/mouse/day demonstrated that L. reuteri 29B did not exert any adverse effects towards the mice which were evident through the absence of pathological changes in the histological examination and blood test. There was also a lack of bacterial translocation as confirmed by Random Amplified Polymorphic DNA (RAPD). The present study highlights the possibility of utilising L. reuteri 29B as a probiotic. It also demonstrates the possibility of isolating potential probiotics from the anogenital region.

Streptococcus agalactiae is one of the most important pathogens infecting tilapia worldwide. In this study, we determined the serotype, sequence type (ST), virulence gene profile and pathogenicity of S. agalactiae isolated from cultured fish. A collection of 256 S. agalactiae that were previously isolated from outbreaks of streptococcosis in red hybrid tilapia in Peninsular Malaysia were studied. The genotype and ST of the isolates were determined by molecular serotyping of the capsular polysaccharide (cps) gene clusters, and multi-locus sequence typing (MLST). The virulence gene profiles were constructed using m-PCR for 14 standard virulence genes with published primers. Two serotypes, with different associated STs, were identified in this study: serotypes Ia ST7 and III ST283. Serotype Ia ST7 lacks lmb, scpB, pavA, fbsB, cyl, bca, cspA and bac genes, which were present in serotype III ST283. The LD50–240h of S. agalactiae in red hybrid tilapia for serotypes Ia ST7 and III ST283 were 8.7 × 103 CFU/mL and 6.3 × 103 CFU/mL, respectively. The histopathological findings were consistent with meningitis, necrosis of hepatocytes, coagulative tubular necrosis and hypocellular of the spleen. Histopathological scores revealed significant differences (p

Introduction: Streptococcus pneumoniae remains as a major causative agent for morbidity and mortality cases in the world. The pathogenicity of this bacterium is attributed to many virulence factors. These factors, though not all had been extensively investigated at the molecular level in various mutant studies. However, of these studies, none of them were using local Malaysian isolate. Materials and Methods: Therefore, to better understand the growth pattern of local mutants a growth curve study and optochin disk susceptibility test were conducted. Several virulence genes (i.e hyl, ugl, ccpA, nanA, and ply) were targeted for this local mutant construction. Pneumococcal mutants were constructed by employing genome-wide deletion technique using high throughput polymerase chain reaction (PCR). Linear knockout construct was established by fusing the kanamycin resistance protein encoding gene, aphA-3 with 1 Kb homologous sequence up and down stream of the respective targeted genes. The gene was knocked out by allelic replacement mechanism. Results: Glycosaminoglycan (GAG) gene cluster mutants (∆hyl- and ∆ugl-) took two days compared to 3 others to grow on brain heart infusion (BHI)-kanamycin agar plate following transformation. This indicated a variance in the mutant growth pattern. Therefore, a growth curve study was conducted to validate the previous observation. We report different time span of log phase in growth curve pattern of mutants relative to the wild type strain. Meanwhile, comparison of zone of inhibition using optochin disk showed non-identical inhibition diameter (≥ 14mm) corresponding to the wild type (14.3mm). Conclusion: The difference in growth pattern was attributed to nutritional role of certain gene.