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Next generation sequencing technologies has revolutionized genomic research by producing a large volume of sequence data and lowest per base cost compared to the traditional sanger method. Although this technology offers many advantages, gap occurrences are commonly found in draft assemblies. The same problem was observed with Rat Cytomegalovirus (RCMV) ALL-03 (Malaysia strain), where a complete genome sequence could not produce the complete genome due to the presence of gaps in the draft genome. This restrains our ability to take full advantage of genome data. This study aimed to identify the sequence data present in the gap regions and close these gaps in order to produce a complete genome sequence for RCMV ALL-03. Twenty sets of specific primers were designed between two adjacent contigs and PCR was carried out to obtain the appropriate sequences in respective gap regions. Sanger sequencing was employed in the PCR product to get the gap sequences. Out of the five identified gaps in the RCMV ALL-03 genome sequence, only three were confirmed to be true gaps, while the other two were due to sequence repeats. In conclusion, all the gaps were closed successfully and complete genome sequence of RCMV ALL-03 can now be explored in further studies.

Pasteurella multocida was first shown to be the causative agent of fowl cholera by Louis Pasteur in 1881. Since then, this Gramnegative bacterium has been identified as the causative agent of many other economically important diseases in a wide range of hosts. Haemorrhagic Septicaemia (HS) is a disease, caused by Pasteurella multocida serotypes B2 and E2 that causes acute, highly fatal septicaemia with high morbidity and mortality in cattle and more susceptible in buffaloes. In this review, we aim to determine the host cell and antibody response towards Pasteurella multicoda B2 infection causing Haemorrhagic septicaemia.

Cytomegalovirus (CMV) is one of the most commonly studied members of the Herpesviridae family. So far several strains have been identified but human CMV (HCMV) remains the most important due to its ability to infect humans and also cross the placenta causing neonatal infection and mortality. However, there are several limitations in the study of this strain which includes availability and use of model animals. Although strains such as Rhesus CMV (RhCMV), Guinea pig CMV (GPCMV) and Rat CMV (RCMV ALL-03) have been shown to cross the placenta and exhibit similar pathogenesis to HCMV, only RCMV ALL-03 can be conveniently used without much limitations as guinea pigs are resistant to antiviral drugs, while rhesus monkeys are seropositive, considered costly and have ethical use restrictions. We therefore propose the use of RCMV ALL-03 as a model for the study of CMV infection.

The gram-negative bacterium Pasteurella multocida serotype B: 2 is an important veterinary and opportunistic human pathogen that causes deplorable welfare problems and colossal economic production losses in bovine and other animal species throughout the world. Pasteurella multocida type B: 2 and its LPS are associated with Haemorrhagic Septicaemia (HS) in mice as animal model. Sixty healthy Balb c mice were placed in twelve plastic cages each one containing five mice. The mice were divided into three major groups (A, B and C). Group A is the control group (n = 10) and these were inoculated with 0.4 mL of PBS pH 7.4 orally. The treatment groups (B; n = 25 and C; n = 25) were inoculated with P. multocida type B: 2 and lipopolysaccharide respectively. The mice in group B and C were further divided into five subgroups. The subgroups were designated based on the graded doses as B101, B103, B105, B107 and B109 for Pasteurella multocida and C101, C103, C105, C107 and C109 for LPS respectively. The mice were observed for 120 h post-inoculation. The concentration of IL-1I² was significantly higher (p

Oral infections of mice as animal model with Pasteurella multocida type B: 2 and its lipopolysaccharides is liable to cause detrimental alterations in the pituitary glands, which results in disruption of reproductive hormonal levels as well as organ functions in the male and female animals; thereby impeding the fertility of at risk animal population. Therefore, this study was designed to evaluate the histopathological modifications in the pituitary glands, reproductive organs and the associated hormonal levels in male and female mice inoculated orally with P. multocida type B: 2 and its lipopolysaccharides. Forty eight mice were divided into three groups consisting of sixteen mice each (8 male and 8 female in separate cages). Group 1 served as the control group and were inoculated orally with 0.4 mL Phosphate Buffer Saline (PBS), group 2 were inoculated orally with 0.4 mL of 109 Colony Forming Unit (CFU) of P. multocida type B: 2 while group 3 were inoculated orally with 0.4 mL of LPS from 109 of colony P. multocida type B: 2. There were significant differences (p

Haemorrhagic Septicaemia (HS) is a fatal systemic disease of bovines typified by an acute, highly fatal septicaemic disease with high morbidity and mortality. In this study, sixty five adult ICR mice were used for the study. In the first phase, 5 mice each were inoculated with 109 cfu of P. multocida type B:2 through the intraperitoneal route. After development of classical signs of HS, the mice were euthanised and dropped into a tank of river waterfor 24, 48 and 72 h. Contaminated water from the tank were collected at the three different times (24, 48 and 72 h) and was used for intraperitoneal, oral and aerosol inoculation of 15 mice each with a dose of 1 mL-1 109 cfu (intraperitoneal and aerosol) and 0.4 mL-1 109cfu (oral). The control group (n = 5) was given phosphate buffered saline. All mice in the three groups and control were culled after 72 h post inoculation and the vital organs (liver, spleen, lung, kidney, heart, brain) were collected in 10% buffered formalin, procesed, sectioned and stained with H&E. Degeneration and necrosis, hemorrhage and infiltration of inflammatory cells were the most observed lesions in all the organs, while thrombosis and oedema were least observed. The lesion severity was moderate to severe using contaminated water at 72 h post contamination and mild to moderate when inoculated with water 24 and 48 h post contamination. Mice inoculated through the oral and intraperitoneal routes recorded more severe lesion compared with those inoculated through the aerosol route. However, using contaminated water at 24 h, mice inoculated through the aerosol route had the highest distribution of inflammatory cells in the lungs, which declined following inoculation with contaminated water at 48 and 72 h, respectively.This study has shown that contamination of water bodiesby carcasses of HS infected animals is a likely source of infection to heathy animals in the wild and that development of lesions severity is time dependant and significantly higher using contaminated water at 72 h than water contaminated for 24 and 48 h. This further indicates the danger of leaving decomposing carcasses in water bodies for prolonged periods.

This study was conducted in order to determine the prevalence and bacteriological assessment of subclinical mastitis and antimicrobial resistance of bacterial isolates from dairy cows in different farms around Selangor, Malaysia. A total of 120 milk samples from 3 different farms were randomly collected and tested for subclinical mastitis using California Mastitis Test (CMT), as well as for bacterial culture for isolation, identification and antimicrobial resistance. The most prevalent bacteria was Staphylococcus sp. (55%), followed by Bacillus sp., (21%) and Corynebacterium sp., (7%), Yersinia sp. and Neisseria sp. both showed 5% prevalence, other species with prevalence below 5% are Acinetobacter sp., Actinobacillus sp., Vibrio sp., Pseudomonas sp., E.coli, Klebsiella sp. and Chromobacter sp. Selected Staphylococcus sp. showed a mean antimicrobial resistance of 73.3% to Ampicillin, 26.7% to Penicillin, Methicillin and Compound Sulphonamide each, 20% to Oxacillin, Amoxycillin and Cefuroxime, 13.3% to Polymyxin B, Erythromycin, Ceftriaxone and Azithromycin and 6.7% to Streptomycin, Clindamycin, Lincomycin and Tetracycline each. This study indicates the need for urgent and effective control measures to tackle the increase in prevalence of subclinical mastitis and their antimicrobial resistance in the study area.