The increased occurrence of Salmonella occurrence in local indigenous vegetables and poultry meat can be a potential health hazards. This study is aimed to detect the prevalence of twenty different virulence factors among Salmonella enterica strains isolated from poultry and local indigenous vegetables in Malaysia via an optimized, rapid and specific multiplex PCR assay. The assay encompasses a total of 19 Salmonella pathogenicity islands genes and a quorum sensing gene (sdiA) in three multiplex reaction sets. A total of 114 Salmonella enterica isolates belonging to 38 different serovars were tested. Each isolate in under this study was found to possess up to 70% of the virulence genes tested and exhibited variable pathogenicity gene patterns. Reproducibility of the multiplex PCR assay was found to be 100% and the detection limit of the optimized multiplex PCR was tested with lowest detectable concentration of DNA 0.8 pg microl(-1). This study demonstrated various Salmonella pathogenicity island virulence gene patterns even within the same serovar. This sets of multiplex PCR system provide a fast and reliable typing approach based on Salmonella pathogenicity islands, thus enabling an effective monitoring of emerging pathogenic Salmonella strains as an additional tool in Salmonella surveillance studies.
Forty-eight strains of Salmonella enterica subsp. enterica serovar Agona and 33 strains of Salmonella enterica subsp. enterica serovar Weltevreden were characterized by random amplified polymorphic DNA (RAPD) fingerprinting using 3 different arbitrary primer, Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR) and antimicrobial susceptibility testing. By using RAPD, 81 strains (44 strains of S. Agona and 33 strains of S. Weltevreden) can be clustered into 14 groups and 6 single isolates whereas ERIC-PCR produced 7 clusters and 3 single isolates. Thirteen antimicrobial agents were used and all the isolates were resistant to erythromycin and showed Multiple Antimicrobial Resistance indexes, ranging from 0.08 to 0.62. Poultry still remain as the common reservoir for multi-drug-resistant Salmonella. On the other hand, vegetables contaminated with S. Weltevreden showed a gain in antimicrobial resistance. Besides that, consistent antibiograms were observed from S. Weltevreden isolated at Kajang wet market on 2000/08/02.
After an intracellular calcium influx, fusion of the sperm plasma membrane and outer acrosomal membrane (the acrosome reaction) precedes mammalian fertilization in vivo. This study describes the isolation of outer acrosomal membrane from ram spermatozoa and the subsequent characterization of calcium-binding proteins. Pooled ejaculates were diluted, cooled slowly and washed. Incubation with Hyamine 1622 (benzethenium chloride) and subsequent slow centrifugation gently dislodged and concentrated acrosomal membranes, the fragments of which were isolated on a two-step discontinuous sucrose gradient. The acrosomal membrane material stained with Giemsa, whereas spermatozoa from the gradient pellet stained intensely only in the equatorial segment. The acrosomal fraction showed a limited number of polypeptides by SDS-PAGE. Incubation with 45Ca2+ revealed two radioactive bands at 34 and 39 kDa. Extraction in the presence of EGTA implied that these proteins are not peripheral proteins associated with the membrane only in the presence of calcium ions, but are integral membrane proteins. Polyclonal antisera raised to the two bands showed specific binding to the anterior acrosomal region and demonstrated the intracellular location of the proteins. Sequence data of protein A revealed 83% homology with calnexin homologue precursor and 70% homology with annexin XI. Protein B showed 68% homology with protein SP-10 precursor and 64-72% homology with various annexins. However, crossreactivity with a range of commercial annexin antibodies and a specific antibody to a synthetic motif encompassing the annexin calcium-binding site was not demonstrable. It is concluded that the isolated proteins are unlikely to be annexins, but are possibly novel calcium-binding protein
Testes from nine male Malin x Santa-Ines rams with an average body weight of 43.1+/-3.53 kg, were used to study the effects of palm kernel cake (PKC) based diet on spermatogenic cells and to assess copper (Cu) levels in liver, testis and plasma in sheep. Animals were divided into three groups and randomly assigned three dietary treatments using restricted randomization of body weight in completely randomized design. The dietary treatments were 60% palm kernel cake plus 40% oil palm frond (PKC), 60% palm kernel cake plus 40% oil palm frond supplemented with 23 mg/kg dry matter of molybdenum as ammonium molybdate [(NH(4))(6)Mo(7)O(24).4H(2)O] and 600 mg/kg dry matter of sulphate as sodium sulphate [Na(2)SO(4)] (PKC-MS) and 60% concentrate of corn-soybean mix+40% oil palm frond (Control), the concentrate was mixed in a ratio of 79% corn, 20% soybean meal and 1% standard mineral mix. The results obtained showed that the number of spermatogonia, spermatocytes, spermatids and Leydig cells were not significantly different among the three treatment groups. However, spermatozoa, Sertoli cells and degenerated cells showed significant changes, which, may be probably due to the Cu content in PKC. Liver and testis Cu levels in the rams under PKC diet was found to be significantly higher (P<0.05) than rams in Control and PKC-MS diets. Plasma Cu levels showed a significant increase (P<0.05) at the end of the experiment as compared to at the beginning of the experiment for PKC and Control. In conclusion, spermatogenesis is normal in rams fed the diet without PKC and PKC supplemented with Mo and S. However spermatogenesis was altered in the PKC based diet probably due to the toxic effects of Cu and the significant changes in organs and plasma. Thus, Mo and S play a major role in reducing the accumulation of Cu in organs.
Introduction: Pheromones are chemicals produced by an animal that affects the behavior of another animal or the same species. Information conveyed includes location, presence of food or threat, sexual attraction, courtship, and dam-pup interactions. Pheromones are used widely in laboratory mice facilities to synchronize estrus and simultaneous breeding for logistic purposes. Female mice housed together in the absence of the male exhibit the Lee-Boot effect of lengthened diestrus or ovarian inactive period of up to several weeks. Whitten effect is described when a large number of female mice housed together in the absence of the male and having diestrus, will enter estrous 48 to 72 hours later upon exposure to male odors or male mouse urine soaked-bedding. Objective: The aim of this study is to determine the time taken for the Whitten effect to occur based on changes in vaginal cell characteristics, vulva appearance and behavior in grouped female mice. Methodology: Ten female mice were acclimatized to the animal facility for 3 estrus cycles or 12 days. Phases of the estrus cycle were evaluated by visual observation to assess changes to the vulva and vaginal cytology. Male urine soaked-bedding were exposed to females for 3 days and the time taken for the Whitten effect to occur was determined based on changes in vaginal cell characteristics, vulva appearance and observation of behavior. Result: The Cochran's Q test was used to observe the changes from diestrus to proestrus and later estrus. The results showed a significant difference (p<0.05) in the number of mice that successfully enter the proestrus and estrus phases over a four time point, χ 2 (2) = 18.857. Conclusion: The Whitten effect occurs after 72 hours of exposure to male urine soaked-bedding based on vulva appearance, vaginal cell characteristics and behavior in grouped female mice.
Asian Conference on Biomedical Research and Laboratory Medicine was organized by three associations: Malaysian Association of Clinical Biochemists (MACB), Association of Scientific Officers Ministry of Health (ASOMH), and Malaysian Biomedical Science Association (MyBIOMED). The conference was held on 24th-25th August 2021 and a workshop on 26th August 2021 through Webex Webinar Platform with the theme of “Revolutionising laboratory medicine through research and innovation”. This conference aimed to bring together leading academic, scientists, laboratory medical scientists, pathologists, researchers, and research scholars to exchange and share their experiences and research results about all aspects of laboratory medicine and health sciences. There were a total of almost 500 participants registered and over 100 abstracts submitted for this conference.
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