Previous studies have identified differential expression of immune-mediated genes related to infl... more Previous studies have identified differential expression of immune-mediated genes related to inflammatory response in chickens with different genetic susceptibility to IBDV infections. However, the mechanisms of genetic resistance against IBD are not known. RNA sequencing through NGS technologies provide an excellent platform to study differentially expressed genes of known or unknown functions to better define mechanisms of host resistance. Therefore, this study aimed at investigating susceptibility of different inbred chicken lines toward very virulent IBDV through transcriptomic analysis. This analysis allows for quantification and comparison of gene expression. Bioinformatics analysis of this data will allow function annotation of differentially expressed genes, indicating possible roles in the response to infection. Gene of interest, virus load detection, indel, SNP and CNV between different lines and breeds will be validated using qPCR. This study is expected to provide inform...
In Malaysia, nest of Aerodramus fuciphagus (white-nest swiftlet) and Aerodramus maximus (black-ne... more In Malaysia, nest of Aerodramus fuciphagus (white-nest swiftlet) and Aerodramus maximus (black-nest swiftlet) are harvested for commercial purposes as one of most valuable animal product. Swiftlet taxonomy has been controversial due to numerous undefined parameters. Morphological differences between swiftlet species from different habitats remain unclear. This study found that A. fuciphagus from natural habitat are generally larger in size compared to man-made habitat and A. maximus are larger compared to A. fuciphagus. We postulated the different in body size is due to dietary behavior of the swiftlets.
Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known ... more Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known as B00/81) and UPM190 (also known as UPM04/190) isolated from local IBD outbreaks in 2000 and 2004, respectively, were separately passaged for 12 consecutive times in 11-day-old specific pathogen free (SPF) chicken embryonated eggs (CEE) via the chorioallantoic membrane (CAM) route. The CEE passage 8 (EP8) isolates were passaged once in BGM-70 cell line yielding UPM0081EP8BGMP1 and UPM190EP8BGMP1, while the EP12 isolates were passaged 15 times in BGM-70 cell line yielding UPM0081EP12BGMP15 and UPM190EP12BGMP15 using T25 tissue culture flask. These isolates were all propagated once in bioreactor using cytodex 1 as microcarrier at 3 g per liter (3 g/L) yielding UPM0081EP8BGMP1BP1, UPM190EP8BGMP1BP1, UPM0081EP12BGMP15BP1, and UPM190EP12BGMP15BP1 isolates. The viruses were harvested at 3 days after inoculation, following the appearance of cytopathic effects (CPE) characterized by detachment ...
Background DNA vaccines offer several advantages over conventional vaccines in the development of... more Background DNA vaccines offer several advantages over conventional vaccines in the development of effective vaccines against avian influenza virus (AIV). However, one of the limitations of the DNA vaccine in poultry is that it induces poor immune responses. In this study, chicken interleukin (IL) -15 and IL-18 were used as genetic adjuvants to improve the immune responses induced from the H5 DNA vaccination in chickens. The immunogenicity of the recombinant plasmid DNA was analyzed based on the antibody production, T cell responses and cytokine production, following inoculation in 1-day-old (Trial 1) and 14-day-old (Trial 2) specific-pathogen-free chickens. Hence, the purpose of the present study was to explore the role of chicken IL-15 and IL-18 as adjuvants following the vaccination of chickens with the H5 DNA vaccine. Results The overall HI antibody titer in chickens immunized with pDis/H5 + pDis/IL-15 was higher compared to chickens immunized with pDis/H5 (p < 0.05). The find...
A NDV isolate designated as IBS002 was isolated from the commercial broiler farm in Malaysia. The... more A NDV isolate designated as IBS002 was isolated from the commercial broiler farm in Malaysia. The virus was characterized as a virulent strain based on the multiple basic amino acid motif of the F cleavage site (112)RRRKGF(117) and the length of C-terminus extension of the HN gene. Besides that, IBS002 also was classified as a velogenic NDV with MDT of 51.2 hours and ICPI of 1.76. The genetic distance analysis based on the full-length of F and HN genes showed that both the velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had higher genetic variations with the genotype II LaSota vaccine; in comparison to the recombinant genotype VII vaccine This study also evaluated the protection efficacy of the recombinant genotype VII NDV inactivated vaccine when added to an existing commercial vaccination program against challenged with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The result indicated that both LaSota a...
Genotype VII Newcastle disease viruses are associated with huge economic losses in the global pou... more Genotype VII Newcastle disease viruses are associated with huge economic losses in the global poultry industry. Despite the intensive applications of vaccines, disease outbreaks caused by those viruses continue to occur frequently even among the vaccinated poultry farms. An important factor in the suboptimal protective efficacy of the current vaccines is the genetic mismatch between the prevalent strains and the vaccine strains. Therefore, in the present study, an effective and stable genotype-matched live attenuated Newcastle disease virus (NDV) vaccine was developed using reverse genetics, based on a recently isolated virulent naturally recombinant NDV IBS025/13 Malaysian strain. First of all, the sequence encoding the fusion protein (F) cleavage site of the virus was modified in silico from virulent polybasic (RRQKRF) to avirulent monobasic (GRQGRL) motif. The entire modified sequence was then chemically synthesized and inserted into pOLTV5 transcription vector for virus rescue. ...
Vaccination is an essential component in controlling infectious bursal disease (IBD), however, th... more Vaccination is an essential component in controlling infectious bursal disease (IBD), however, there is a lack of information on the genetic characteristics of a recent infectious bursal disease virus (IBDV) that was isolated from IBD vaccinated commercial flocks in Malaysia. The present study investigated 11 IBDV isolates that were isolated from commercial poultry farms. The isolates were detected using reverse transcription-polymerase chain reaction (RT-PCR) targeting the hypervariable region (HVR) of VP2. Based on the HVR sequences, five isolates (IBS536/2017, IBS624/2017, UPM766/2018, UPM1056/2018, and UPM1432/2019) were selected for whole-genome sequencing using the MiSeq platform. The nucleotide and amino acid (aa) sequences were compared with the previously characterized IBDV strains. Deduced aa sequences of VP2HVR revealed seven isolates with 94–99% aa identity to very virulent strains (genogroup 3), two isolates with 97–100% aa identity to variant strains (genogroup 2), and...
Newcastle disease (ND) is one of the most important avian diseases with considerable threat to th... more Newcastle disease (ND) is one of the most important avian diseases with considerable threat to the productivity of poultry all over the world. The disease is associated with severe respiratory, gastrointestinal, and neurological lesions in chicken leading to high mortality and several other production related losses. The aetiology of the disease is an avian paramyxovirus type-1 or Newcastle disease virus (NDV), whose isolates are serologically grouped into a single serotype but genetically classified into a total of 19 genotypes, owing to the continuous emergence and evolution of the virus. In Nigeria, molecular characterization of NDV is generally very scanty and majorly focuses on the amplification of the partial F gene for genotype assignment. However, with the introduction of the most objective NDV genotyping criteria which utilize complete fusion protein coding sequences in phylogenetic taxonomy, the enormous genetic diversity of the virus in Nigeria became very conspicuous. In...
Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (... more Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 10 TCID/mL and log10 TCID/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogen...
Fowl adenovirus (FAdV) is distributed worldwide and caused economic losses in poultry industry. T... more Fowl adenovirus (FAdV) is distributed worldwide and caused economic losses in poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate of Malaysia in specific pathogen free (SPF) chicken embryonated eggs (CEE) and its infectivity in commercial broiler chickens. SPF CEE was inoculated with 0.1mL FAdV inoculum via chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20) with virus titer of 10TCID/mL was inoculated (0.5mL) into day old commercial broiler chickens either via oral or intraperitoneal route. The study demonstrated that 100% embryonic mortality was recorded from passage 2 (E2) to E20 with delayed pattern at passage 17 onwards. The lesions were confined in the liver and CAM. Substitutions of amino acids in L1 loop of hexon at position 49 and 66, and in knob of fiber gene at position 318 and 322 were recorded in the E20 isolate. The isolate is belong to serotype 8b and no...
Salmonella enterica subsp. enterica serovar Typhimurium is one of several well-categorized Salmon... more Salmonella enterica subsp. enterica serovar Typhimurium is one of several well-categorized Salmonella serotypes recognized globally. Here, we report the whole-genome sequence of S Typhimurium strain UPM 260, isolated from a broiler chicken.
Previous studies have identified differential expression of immune-mediated genes related to infl... more Previous studies have identified differential expression of immune-mediated genes related to inflammatory response in chickens with different genetic susceptibility to IBDV infections. However, the mechanisms of genetic resistance against IBD are not known. RNA sequencing through NGS technologies provide an excellent platform to study differentially expressed genes of known or unknown functions to better define mechanisms of host resistance. Therefore, this study aimed at investigating susceptibility of different inbred chicken lines toward very virulent IBDV through transcriptomic analysis. This analysis allows for quantification and comparison of gene expression. Bioinformatics analysis of this data will allow function annotation of differentially expressed genes, indicating possible roles in the response to infection. Gene of interest, virus load detection, indel, SNP and CNV between different lines and breeds will be validated using qPCR. This study is expected to provide inform...
In Malaysia, nest of Aerodramus fuciphagus (white-nest swiftlet) and Aerodramus maximus (black-ne... more In Malaysia, nest of Aerodramus fuciphagus (white-nest swiftlet) and Aerodramus maximus (black-nest swiftlet) are harvested for commercial purposes as one of most valuable animal product. Swiftlet taxonomy has been controversial due to numerous undefined parameters. Morphological differences between swiftlet species from different habitats remain unclear. This study found that A. fuciphagus from natural habitat are generally larger in size compared to man-made habitat and A. maximus are larger compared to A. fuciphagus. We postulated the different in body size is due to dietary behavior of the swiftlets.
Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known ... more Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known as B00/81) and UPM190 (also known as UPM04/190) isolated from local IBD outbreaks in 2000 and 2004, respectively, were separately passaged for 12 consecutive times in 11-day-old specific pathogen free (SPF) chicken embryonated eggs (CEE) via the chorioallantoic membrane (CAM) route. The CEE passage 8 (EP8) isolates were passaged once in BGM-70 cell line yielding UPM0081EP8BGMP1 and UPM190EP8BGMP1, while the EP12 isolates were passaged 15 times in BGM-70 cell line yielding UPM0081EP12BGMP15 and UPM190EP12BGMP15 using T25 tissue culture flask. These isolates were all propagated once in bioreactor using cytodex 1 as microcarrier at 3 g per liter (3 g/L) yielding UPM0081EP8BGMP1BP1, UPM190EP8BGMP1BP1, UPM0081EP12BGMP15BP1, and UPM190EP12BGMP15BP1 isolates. The viruses were harvested at 3 days after inoculation, following the appearance of cytopathic effects (CPE) characterized by detachment ...
Background DNA vaccines offer several advantages over conventional vaccines in the development of... more Background DNA vaccines offer several advantages over conventional vaccines in the development of effective vaccines against avian influenza virus (AIV). However, one of the limitations of the DNA vaccine in poultry is that it induces poor immune responses. In this study, chicken interleukin (IL) -15 and IL-18 were used as genetic adjuvants to improve the immune responses induced from the H5 DNA vaccination in chickens. The immunogenicity of the recombinant plasmid DNA was analyzed based on the antibody production, T cell responses and cytokine production, following inoculation in 1-day-old (Trial 1) and 14-day-old (Trial 2) specific-pathogen-free chickens. Hence, the purpose of the present study was to explore the role of chicken IL-15 and IL-18 as adjuvants following the vaccination of chickens with the H5 DNA vaccine. Results The overall HI antibody titer in chickens immunized with pDis/H5 + pDis/IL-15 was higher compared to chickens immunized with pDis/H5 (p < 0.05). The find...
A NDV isolate designated as IBS002 was isolated from the commercial broiler farm in Malaysia. The... more A NDV isolate designated as IBS002 was isolated from the commercial broiler farm in Malaysia. The virus was characterized as a virulent strain based on the multiple basic amino acid motif of the F cleavage site (112)RRRKGF(117) and the length of C-terminus extension of the HN gene. Besides that, IBS002 also was classified as a velogenic NDV with MDT of 51.2 hours and ICPI of 1.76. The genetic distance analysis based on the full-length of F and HN genes showed that both the velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had higher genetic variations with the genotype II LaSota vaccine; in comparison to the recombinant genotype VII vaccine This study also evaluated the protection efficacy of the recombinant genotype VII NDV inactivated vaccine when added to an existing commercial vaccination program against challenged with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The result indicated that both LaSota a...
Genotype VII Newcastle disease viruses are associated with huge economic losses in the global pou... more Genotype VII Newcastle disease viruses are associated with huge economic losses in the global poultry industry. Despite the intensive applications of vaccines, disease outbreaks caused by those viruses continue to occur frequently even among the vaccinated poultry farms. An important factor in the suboptimal protective efficacy of the current vaccines is the genetic mismatch between the prevalent strains and the vaccine strains. Therefore, in the present study, an effective and stable genotype-matched live attenuated Newcastle disease virus (NDV) vaccine was developed using reverse genetics, based on a recently isolated virulent naturally recombinant NDV IBS025/13 Malaysian strain. First of all, the sequence encoding the fusion protein (F) cleavage site of the virus was modified in silico from virulent polybasic (RRQKRF) to avirulent monobasic (GRQGRL) motif. The entire modified sequence was then chemically synthesized and inserted into pOLTV5 transcription vector for virus rescue. ...
Vaccination is an essential component in controlling infectious bursal disease (IBD), however, th... more Vaccination is an essential component in controlling infectious bursal disease (IBD), however, there is a lack of information on the genetic characteristics of a recent infectious bursal disease virus (IBDV) that was isolated from IBD vaccinated commercial flocks in Malaysia. The present study investigated 11 IBDV isolates that were isolated from commercial poultry farms. The isolates were detected using reverse transcription-polymerase chain reaction (RT-PCR) targeting the hypervariable region (HVR) of VP2. Based on the HVR sequences, five isolates (IBS536/2017, IBS624/2017, UPM766/2018, UPM1056/2018, and UPM1432/2019) were selected for whole-genome sequencing using the MiSeq platform. The nucleotide and amino acid (aa) sequences were compared with the previously characterized IBDV strains. Deduced aa sequences of VP2HVR revealed seven isolates with 94–99% aa identity to very virulent strains (genogroup 3), two isolates with 97–100% aa identity to variant strains (genogroup 2), and...
Newcastle disease (ND) is one of the most important avian diseases with considerable threat to th... more Newcastle disease (ND) is one of the most important avian diseases with considerable threat to the productivity of poultry all over the world. The disease is associated with severe respiratory, gastrointestinal, and neurological lesions in chicken leading to high mortality and several other production related losses. The aetiology of the disease is an avian paramyxovirus type-1 or Newcastle disease virus (NDV), whose isolates are serologically grouped into a single serotype but genetically classified into a total of 19 genotypes, owing to the continuous emergence and evolution of the virus. In Nigeria, molecular characterization of NDV is generally very scanty and majorly focuses on the amplification of the partial F gene for genotype assignment. However, with the introduction of the most objective NDV genotyping criteria which utilize complete fusion protein coding sequences in phylogenetic taxonomy, the enormous genetic diversity of the virus in Nigeria became very conspicuous. In...
Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (... more Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 10 TCID/mL and log10 TCID/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogen...
Fowl adenovirus (FAdV) is distributed worldwide and caused economic losses in poultry industry. T... more Fowl adenovirus (FAdV) is distributed worldwide and caused economic losses in poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate of Malaysia in specific pathogen free (SPF) chicken embryonated eggs (CEE) and its infectivity in commercial broiler chickens. SPF CEE was inoculated with 0.1mL FAdV inoculum via chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20) with virus titer of 10TCID/mL was inoculated (0.5mL) into day old commercial broiler chickens either via oral or intraperitoneal route. The study demonstrated that 100% embryonic mortality was recorded from passage 2 (E2) to E20 with delayed pattern at passage 17 onwards. The lesions were confined in the liver and CAM. Substitutions of amino acids in L1 loop of hexon at position 49 and 66, and in knob of fiber gene at position 318 and 322 were recorded in the E20 isolate. The isolate is belong to serotype 8b and no...
Salmonella enterica subsp. enterica serovar Typhimurium is one of several well-categorized Salmon... more Salmonella enterica subsp. enterica serovar Typhimurium is one of several well-categorized Salmonella serotypes recognized globally. Here, we report the whole-genome sequence of S Typhimurium strain UPM 260, isolated from a broiler chicken.
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