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Infectious Bronchitis (IB) is an economically important avian disease that considerably threatens the global poultry industry. This is partly, as a result of its negative consequences on egg production, weight gain as well as mortality rate.The disease is caused by a constantly evolving avian infectious bronchitis virus whose isolates are classified into several serotypes and genotypes that demonstrate little or no cross protection. In order to curb the menace of the disease therefore, broad based vaccines are urgently needed. The aim of this study was to develop a recombinant DNA vaccine candidate for improved protection of avian infectious bronchitis in poultry. Using bioinformatics and molecular cloning procedures, sets of monovalent and bivalent DNA vaccine constructs were developed based on the S1 glycoprotein from classical and variants IBV strains namely, M41 and CR88 respectively. The candidate vaccine was then encapsulated with a chitosan and saponin formulated nanoparticle for enhanced immunogenicity and protective capacity. RT-PCR assay and IFAT were used to confirm the transcriptional and translational expression of the encoded proteins respectively, while ELISA and Flow-cytometry were used to evaluate the immunogenicity of the candidate vaccine following immunization of various SPF chicken groups (A-F). Furthermore, histopathological changes and virus shedding were determined by quantitative realtime PCR assay and lesion scoring procedure respectively following challenge of various subgroups with respective wild-type IBV viruses. Results obtained from this study showed that, groups vaccinated with a bivalent DNA vaccine construct (pBudCR88-S1/M41-S1) had a significant increase in anti-IBV antibodies, CD3+ and CD8+ T-cells responses as compared to non-vaccinated groups. Likewise, the bivalent vaccine candidate significantly decreased the oropharyngeal and cloacal virus shedding (p < 0.05) compared to non-vaccinated control. Chickens immunized with the bi

Infectious Bronchitis (IB) is an economically important avian disease that considerably threatens the global poultry industry. This is partly, as a result of its negative consequences on egg production, weight gain as well as mortality rate.The disease is caused by a constantly evolving avian infectious bronchitis virus whose isolates are classified into several serotypes and genotypes that demonstrate little or no cross protection. In order to curb the menace of the disease therefore, broad based vaccines are urgently needed. The aim of this study was to develop a recombinant DNA vaccine candidate for improved protection of avian infectious bronchitis in poultry. Using bioinformatics and molecular cloning procedures, sets of monovalent and bivalent DNA vaccine constructs were developed based on the S1 glycoprotein from classical and variants IBV strains namely, M41 and CR88 respectively. The candidate vaccine was then encapsulated with a chitosan and saponin formulated nanoparticle for enhanced immunogenicity and protective capacity. RT-PCR assay and IFAT were used to confirm the transcriptional and translational expression of the encoded proteins respectively, while ELISA and Flow-cytometry were used to evaluate the immunogenicity of the candidate vaccine following immunization of various SPF chicken groups (A-F). Furthermore, histopathological changes and virus shedding were determined by quantitative realtime PCR assay and lesion scoring procedure respectively following challenge of various subgroups with respective wild-type IBV viruses. Results obtained from this study showed that, groups vaccinated with a bivalent DNA vaccine construct (pBudCR88-S1/M41-S1) had a significant increase in anti-IBV antibodies, CD3+ and CD8+ T-cells responses as compared to non-vaccinated groups. Likewise, the bivalent vaccine candidate significantly decreased the oropharyngeal and cloacal virus shedding (p < 0.05) compared to non-vaccinated control. Chickens immunized with the bi

Pneumonic pasteurellosis is an economically important infectious disease in the small ruminant industry which causes sudden death and loss for farmers. Nonetheless, this disease is still a common sight in sheep and goats in Malaysia, probably due to the unpopular usage of pasteurellosis vaccine or inappropriate vaccination practices. The aim of this study was designed to classify the severity of pneumonia via the establishment of auscultation scoring method and to quantify the acute phase proteins and heat shock proteins responses from vaccinated and non-vaccinated goats. Goat farms, consist of vaccinated and non-vaccinated farms, were selected in this study: where 15 clinically normal healthy goats and 9 pneumonic goats were selected from vaccinated farms whereas 15 clinically normal healthy goats and 31 pneumonic goats from non-vaccinated farms were selected for this study. Crackle lung sounds were not detected in both vaccinated and non-vaccinated normal goats. However, vaccinated pneumonic goats showed mild crackle lung sound while non-vaccinated pneumonic goats exhibited moderate crackle lung sound. There were significant increases (p < 0.05) in acute phase proteins and heat shock proteins concentrations for the non-vaccinated pneumonic goats group. In this study, conclusion can be made that the vaccinated goats exhibited very mild clinical responses of pneumonia and non-significant biomarker responses compared to the non-vaccinated goats. Thus, vaccination is an effective preventive measure to control pneumonic pasteurellosis and acute phase proteins and heat shock proteins can be considered as future biomarkers in screening and rapid diagnostic method for this particular disease.

Pneumonic pasteurellosis is an economically important infectious disease in the small ruminant industry which causes sudden death and loss for farmers. Nonetheless, this disease is still a common sight in sheep and goats in Malaysia, probably due to the unpopular usage of pasteurellosis vaccine or inappropriate vaccination practices. The aim of this study was designed to classify the severity of pneumonia via the establishment of auscultation scoring method and to quantify the acute phase proteins and heat shock proteins responses from vaccinated and non-vaccinated goats. Goat farms, consist of vaccinated and non-vaccinated farms, were selected in this study: where 15 clinically normal healthy goats and 9 pneumonic goats were selected from vaccinated farms whereas 15 clinically normal healthy goats and 31 pneumonic goats from non-vaccinated farms were selected for this study. Crackle lung sounds were not detected in both vaccinated and non-vaccinated normal goats. However, vaccinated pneumonic goats showed mild crackle lung sound while non-vaccinated pneumonic goats exhibited moderate crackle lung sound. There were significant increases (p < 0.05) in acute phase proteins and heat shock proteins concentrations for the non-vaccinated pneumonic goats group. In this study, conclusion can be made that the vaccinated goats exhibited very mild clinical responses of pneumonia and non-significant biomarker responses compared to the non-vaccinated goats. Thus, vaccination is an effective preventive measure to control pneumonic pasteurellosis and acute phase proteins and heat shock proteins can be considered as future biomarkers in screening and rapid diagnostic method for this particular disease.

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.

Pasteurella multocida was first shown to be the causative agent of fowl cholera by Louis Pasteur in 1881. Since then, this Gram-negative 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.

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.

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-1β was significantly higher (p<0.001) in the B107 of P. multocida type B: 2 and C109 of LPS compared to the control group and the other treatment groups. The concentration of IL-6 was significantly higher (p<0.001) in the B103 cfu of P. multocida type B: 2 and C109 cfu of LPS compared to the control group and the other treatment groups. Interleukin-1β and interleukin-6 evaluated in this study were predominantly associated with HS in varying doses of 101, 103, 105, 107 and 109 in the whole cell and its LPS respectively. Therefore, the target of this study was to investigate on pro-inflammatory response in mice infected with graded doses of Pasteurella multocid

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.

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