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1.
Prev Vet Med ; 212: 105843, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36682258

RESUMO

The oral lesions associated with foot-and-mouth disease (FMD) negatively affect animal behavior, which can adversely impact animal production and welfare. Physical form of a therapeutic diet (TD) can improve the feed intake and wellbeing during the acute phase of FMD. Accordingly, we tested the effect of two physical forms of a previously developed TD on the behavior of calves experimentally infected with FMD virus (FMDV). Crossbred Holstein Friesian male calves of 10-12 months (n = 12) were experimentally infected with a virulent strain of FMDV and were offered a TD enriched with 19% CP and 2.9 Mcal ME/kg for 11 days post-FMDV infection. One group received the TD in mash form (TDM) while the other (n = 6/group) received it in cooked form (TDC). A group of four calves served as uninfected control and were fed TDM. The time spent by the calves on certain behaviours was recorded in a pre-set form from 06:00-18:00 h for 10 days from day 2-11 post-FMDV infection. The data was divided into two sessions. Session 1 (06:00-13:00 h) represented after the offering of TD, while session 2 (13:01-18:00 h) represented the data after offering green fodder. Based on exploratory data analysis, data recorded from day 2-7 post-FMDV infection was included in the final analysis. Linear mixed model was used by fitting treatment, day and their interaction as fixed effects while calf as random effect. Orthogonal contrast was applied by comparing the infected TDM with other two groups. The results revealed that the cooked form of TD improved the ingestion time, resting time, sleeping time and licking time from day 2-7 post-FMDV infection as compared with the infected TDM group. Ingestive behaviour was better in the infected TDC than that of TDM group (p < 0.01). The sleeping time was significantly high in the infected groups as compared to the uninfected TDM group (p < 0.01) till day 6 post-FMDV infection. Daily activities such as licking, standing and resting differed significantly between the infected TDM and TDC groups in session 1, but not in session 2. Urination and defecation did not differ significantly between the infected TDM and TDC groups. It was concluded that cooked form of TD remediated the effects of infection with FMDV as evidenced by improvement in the behaviour of the calves.


Assuntos
Doenças dos Bovinos , Vírus da Febre Aftosa , Febre Aftosa , Animais , Bovinos , Masculino , Comportamento Alimentar , Dieta/veterinária
2.
Gene ; 810: 146085, 2022 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-34843879

RESUMO

Sheeppox virus (SPPV) is responsible for a significant economic loss to sheep husbandry in enzootic regions of Africa, the Middle East, and Asia including the Indian subcontinent. In this study, we present the complete genome sequence of SPPV vaccine strain SPPV-Srin38/00 from India determined by next-generation sequencing (NGS) using Illumina technology. The attenuated Srinagar vaccine strain of SPPV (SPPV-Srin38/00) was developed by serial passaging the virus initially in lamb testes (LT) cells followed by Vero cell line. The SPPV-Srin38/00 virus has a genome size of 150, 103 bp, which encodes for 147 functional putative genes and consists of a central coding region flanked by two identical 2353 bp inverted terminal repeats (ITRs). Comparative phylogenetic analysis based on complete genome sequences of Capripoxviruses formed three distinct groups each for SPPV, GTPV, and LSDV with clustering of SPPV-Srin38/00 strain with SPPV-A strain. Nine ORFs of SPPV-Srin38/00 namely SPPV-Srin_002/SPPV-Srin_155, SPPV-Srin_004/SPPV-Srin_153, SPPV-Srin_009, SPPV-Srin_013, SPPV-Srin_026, SPPV-Srin_132, and SPPV-Srin_136 were found to be fragmented as compared to LSDV, whereas only one ORF (such as SPPV-Srin_136) was found to be fragmented as compared to GTPV. SPPV genomes, including the SPPV-Srin38/00 strain, shared 99.78-99.98% intraspecies nucleotide identity, indicating that SPPV strains have extremely low genetic diversity. The strain shared 96.80-97.08% and 97.11-97.61% nt identity with GTPV and LSDV strains, respectively. Its ORFs 016, 021, 022, 130 and 138 are the least identical ORFs among three species of the genus Capripoxvirus with 72.5-93% aa identity to GTPV and LSDV strains and may be potentially used for differentiation of CaPV species. This study may contribute to a better understanding of the epidemiology and evolution of capripoxviruses as well as the development of specific detection methods, better expression vectors, and vaccines with improved safety and efficacy.


Assuntos
Capripoxvirus/genética , Animais , Capripoxvirus/classificação , Chlorocebus aethiops , Tamanho do Genoma , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , Ovinos , Doenças dos Ovinos/virologia , Células Vero , Sequenciamento Completo do Genoma
3.
Appl Microbiol Biotechnol ; 106(1): 273-286, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34889988

RESUMO

The development of a negative marker vaccine against the foot-and-mouth disease virus (FMDV) will enhance the capabilities to differentiate vaccinated from infected animals and move forward in the progressive control pathway for the control of FMD. Here, we report the development of mutant FMDV of Asia1 with partial deletion of non-structural proteins 3A and 3B and characterization of their infectivity and protection response in the guinea pig model. The deleted FMDV Asia1/IND/63/1972 mutants, pAsiaΔ3A and pAsiaΔ3A3B1 were constructed from the full-length infectious clone pAsiaWT, the viable virus was rescued, and the genetic stability of the mutants was confirmed by 20 monolayer passages in BHK21 cells. The mutant Asia1 viruses showed comparable growth pattern and infectivity with that of AsiaWT in the cell culture. However, the AsiaΔ3A3B1 virus showed smaller plaque and lower virus titer with reduced infectivity in the suckling mice. In guinea pigs, the AsiaΔ3A3B1 virus failed to induce the disease, whereas the AsiaΔ3A virus induced typical secondary lesions of FMD. Vaccination with inactivated Asia1 mutant viruses induced neutralizing antibody response that was significantly lower than that of the parent virus on day 28 post-vaccination (dpv) in guinea pigs (P < 0.05). Furthermore, challenging the vaccinated guinea pigs with the homologous vaccine strain of FMDV Asia1 conferred complete protection. It is concluded that the mutant AsiaΔ3A3B1 virus has the potential to replace the wild-type virus for use as a negative marker vaccine after assessing the vaccine worth attributes in suspension cell and protective efficacy study in cattle.Key points• Deletion mutant viruses of FMDV Asia1, developed by PCR-mediated mutagenesis of NSP 3A and 3B1, were genetically stable.• The growth kinetics and antigenic relatedness of the mutant viruses were comparable with that of the wild-type virus.• Vaccination of guinea pigs with the deletion mutant viruses conferred complete protection upon challenge with the homologous virus.


Assuntos
Vírus da Febre Aftosa , Febre Aftosa , Vacinas Virais , Animais , Anticorpos Neutralizantes , Bovinos , Febre Aftosa/prevenção & controle , Vírus da Febre Aftosa/genética , Cobaias , Camundongos , Sorogrupo , Vacinas Virais/genética
4.
Vet World ; 11(8): 1043-1046, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30250361

RESUMO

Tribolium castaneum is a small and low maintenance beetle that has emerged as a most suitable insect model for studying developmental biology and functional genetic analysis. Diverse population genetic studies have been conducted using Tribolium as the principal model to establish basic facts and principles of inbreeding experiments and response to the selection and other quantitative genetics fundamentals. The advanced molecular genetic studies presently focused on the use of Tribolium as a typical invertebrate model for higher diploid eukaryotes. After a whole genome sequencing of Tribolium, many areas of functional genomics were unraveled, which enabled the use of it in many technical approaches of genomics. The present text reviews the use of Tribolium in techniques such as RNAi, transgenic studies, immune priming, immunohistochemistry, in situ hybridization, gene sequencing for characterization of microRNAs, and gene editing using engineered endonuclease. In contrast to Drosophila, the T. castaneum holds a robust systemic RNAi response, which makes it an excellent model for comparative functional genetic studies.

5.
Vet World ; 11(4): 535-541, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29805222

RESUMO

Copy number variation (CNV) is a phenomenon in which sections of the genome, ranging from one kilo base pair (Kb) to several million base pairs (Mb), are repeated and the number of repeats vary between the individuals in a population. It is an important source of genetic variation in an individual which is now being utilized rather than single nucleotide polymorphisms (SNPs), as it covers the more genomic region. CNVs alter the gene expression and change the phenotype of an individual due to deletion and duplication of genes in the copy number variation regions (CNVRs). Earlier, researchers extensively utilized SNPs as the main source of genetic variation. But now, the focus is on identification of CNVs associated with complex traits. With the recent advances and reduction in the cost of sequencing, arrays are developed for genotyping which cover the maximum number of SNPs at a time that can be used for detection of CNVRs and underlying quantitative trait loci (QTL) for the complex traits to accelerate genetic improvement. CNV studies are also being carried out to understand the evolutionary mechanism in the domestication of livestock and their adaptation to the different environmental conditions. The main aim of the study is to review the available data on CNV and its role in genetic variation among the livestock.

6.
Biologicals ; 49: 39-45, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28754514

RESUMO

In the present investigation, the thermostability of a live attenuated buffalopox vaccine prepared with an indigenous baffalopox virus isolate (BPXV Vij/96) and freeze-dried under conventional lyophilizing conditions is described. Three different stabilizer combinations like LS (lactalbumin hydralysate + sucrose), LHT (lactalbumin hydralysate + Trehalose dihydrate) and TAA (Trehalose dihydrate + l- Alanine + l-Histidine) were used to prepare the vaccine. The study indicated that the LS stabilizer was found to be the stabilizer of choice followed by LHT and TAA for buffalopox vaccine at all temperatures studied. The presence of stabilizers has beneficial influence in preserving the keeping quality of the vaccine. Further, among the diluents used to reconstitute the freeze-dried buffalopox vaccine, double distilled water, 0.85% normal saline solution and phosphate buffer saline were the choice of diluents in that order. However, 1M MgSO4 did not perform well at higher temperatures. Investigation suggests for using LS as a stabilizer for freeze-drying and any of the three diluents except 1MgSO4 for reconstitution of buffalopox vaccine.


Assuntos
Excipientes/química , Vaccinia virus/química , Vacinas Virais/química , Animais , Chlorocebus aethiops , Liofilização , Células Vero
7.
Mol Biol Rep ; 44(3): 281-287, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28551733

RESUMO

The purpose of this study is to develop a novel Reverse Transcriptase Loop-mediated isothermal amplification (RT-LAMP) based assay for in vitro profiling of heat shock protein 70 (Hsp70) in bovine peripheral blood mononuclear cell (PBMC) culture model utilizing the absorbance level of magnesium pyrophosphate-a by-product of LAMP reaction. A set of bovine Hsp70 specific RT-LAMP primers were designed to detect the differential absorbance level of magnesium pyrophosphate by-product which signifies the degree of Hsp70 amplification from cDNA of thermally induced cultured cells at different recovery periods. The study revealed significant (P < 0.05) correlation between absorbance level and the fold change of Hsp70 transcripts at different kinetic intervals of heat stress recovery in bovine PBMC cell culture models. RT-LAMP based absorbance assay can be used as an indicator to measure the degree of bovine Hsp70 transcripts produced during thermal stress and can be used as an alternative to the traditional Real time PCR assay. Developed RT-LAMP assay can be used as a cost-effective method for profiling of bovine HSP70 gene.


Assuntos
Bovinos/metabolismo , Proteínas de Choque Térmico HSP70/análise , Leucócitos Mononucleares/metabolismo , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Células Cultivadas , Expressão Gênica , Proteínas de Choque Térmico HSP70/genética , Técnicas de Amplificação de Ácido Nucleico/economia , RNA Mensageiro
8.
Int J Biometeorol ; 61(5): 931-941, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27853874

RESUMO

Thermotolerance depends mainly on the health and immune status of the animals. The variation in the immune status of the animals may alter the level of tolerance of animals exposed to heat or cold stress. The present study was conducted to investigate the expression profile of two important nucleotide binding and oligomerization domain receptors (NLRs) (NOD1 and NOD2) and their central signalling molecule RIP2 gene during in vitro thermal-stressed bovine peripheral blood mononuclear cells (PBMCs) of native (Sahiwal) and crossbred (Sahiwal X HF) cattle. We also examined the differential expression profile of certain acute inflammatory cytokines in in vitro thermal-stressed PBMC culture among native and its crossbred counterparts. Results revealed that the expression profile of NOD1/2 positively correlates with the thermal stress, signalling molecule and cytokines. Present findings also highlighted that the expression patterns during thermal stress were comparatively superior among indigenous compared to crossbred cattle which may add references regarding the better immune adaptability of Zebu cattle.


Assuntos
Citocinas/genética , Resposta ao Choque Térmico/genética , Leucócitos Mononucleares/metabolismo , Proteína Adaptadora de Sinalização NOD1/genética , Proteína Adaptadora de Sinalização NOD2/genética , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/genética , Animais , Bovinos , Células Cultivadas , Resposta ao Choque Frio/genética , Feminino , Expressão Gênica , RNA Mensageiro/metabolismo , Especificidade da Espécie
10.
J Therm Biol ; 59: 18-25, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27264883

RESUMO

In a tropical country like India, thermal stress is one of the major factors which significantly affects the productivity of dairy cattle. The present study was aimed to identify the effect of heat and cold stress on cell viability, mitogen stimulation indices, nitric oxide production and HSP70 expression in Sahiwal and Holstein crossbred (Frieswal) population in India. The results indicated that the Sahiwal breed can better withstand the effect of heat and cold stress significantly (P<0.05) when compared to the crossbred cattle due to the higher survivability of the Peripheral Blood Mononuclear Cells (PBMCs) and Phytohemagglutinin (PHA-P) mitogen based stimulation indices. The study also revealed the significant differences (P<0.05) in the level of nitric oxide (µM) production amongst the pre and post thermal stressed samples of Sahiwal and Frieswal crossbred samples. Further, the expression of HSP70 was significantly (P<0.05) higher in Sahiwal compared to Frieswal immediately after heat/cold shock to 6h of recovery as indirect ELISA analysis showed gradual rise in the Hsp70 protein concentration (ng/ml) immediately after heat and cold stress (0h) and reached the peak at 6h of recovery. Western blot and immune fluorescent assay results were also corroborated with the findings of indirect ELISA. In Sahiwal cattle the mRNA expression of HSP70 and its protein concentration were higher (P<0.05) during peak summer (44°C) and winter (10°C) as compared to Frieswal cattle. This investigation supports the earlier information on the higher adaptability of indigenous cattle breeds to hot and humid conditions compared to the crossbreds of temperate cattle breeds.


Assuntos
Bovinos/psicologia , Resposta ao Choque Frio , Proteínas de Choque Térmico HSP70/genética , Resposta ao Choque Térmico , Hibridização Genética , Óxido Nítrico/metabolismo , Criação de Animais Domésticos , Animais , Bovinos/genética , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP70/metabolismo , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Estações do Ano
11.
Mol Cell Probes ; 30(3): 174-7, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26872529

RESUMO

A Loop-mediated isothermal amplification (LAMP) assay targeting the highly conserved DNA polymerase gene of capripox virus genome was developed and evaluated for rapid detection of sheep pox and goat pox viruses. The optimized LAMP assay is found specific and sensitive for amplification of target DNA with a diagnostic sensitivity and specificity of 96.6% and 100% respectively compared to quantitative PCR. The detection rate of LAMP, PCR and Q-PCR assays is found to be 81.5%, 67% and 83% respectively. This LAMP assay has the potential for rapid clinical diagnosis and surveillance of sheep pox and goat pox in field diagnostic laboratories.


Assuntos
Capripoxvirus/genética , Capripoxvirus/isolamento & purificação , Doenças das Cabras/virologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções por Poxviridae/veterinária , Doenças dos Ovinos/virologia , Animais , Eletroforese em Gel de Ágar , Cabras/virologia , Infecções por Poxviridae/virologia , Sensibilidade e Especificidade , Ovinos/virologia
12.
Mol Cell Probes ; 29(3): 193-5, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25828693

RESUMO

A loop-mediated isothermal amplification (LAMP) assay targeting DNA Pol gene was optimized and evaluated for the rapid detection of orf virus in clinical samples. The LAMP assay was found to be specific and sensitive. The detection rate of LAMP (89.3%) was better than PCR (67.9%) and comparable to real-time PCR (91.1%) in clinical samples by gel electrophoresis and visual detection methods. This LAMP assay is simple and does not rely upon any special equipment and could be employed in clinical diagnosis and epidemiological survey of orf infection.


Assuntos
Doenças das Cabras/virologia , Técnicas de Diagnóstico Molecular/veterinária , Técnicas de Amplificação de Ácido Nucleico/métodos , Vírus do Orf/genética , Infecções por Poxviridae/virologia , Doenças dos Ovinos/virologia , Animais , DNA Polimerase Dirigida por DNA/genética , Ectima Contagioso , Cabras , Técnicas de Diagnóstico Molecular/métodos , Ovinos
13.
Vet World ; 8(3): 295-300, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27047088

RESUMO

BACKGROUND/AIM: Recent studies have shown that interleukin-15 (IL-15)is a critical factor for the development and proliferation of CD8(+) memory T cells. The aim of present study is to study the role bovine IL-15 (bIL-15)in activation pathway of bovine CD8(+) T cells if any, which will be useful in designing the adjuvant to increase the duration of immunity of the vaccine preparations. MATERIALS AND METHODS: Coding region of bIL-15 (489) was amplified from cDNA of lipopolysaccharide-induced bovine peripheral blood mononuclear cells (PBMCs) using gene specific primers and cloned into pcDNA3.1(+). Mature length of bIL-15 was amplified using gene specific primers and cloned into pET32a for expression studies. Expressed fusion protein was purified using Ni-Nitrilotriacetic acid agarose affinity chromatography and analyzed by SDS-Polyacryamide gel electrophoresis (PAGE) and western blotting. Biological activity of purified protein was analyzed by quantitative Polymerase Chain Reaction (qPCR) for an increase in levels of Bcl2, STAT3 and STAT5a using cDNA synthesized from RNA of PBMCs induced with different concentrations of purified bIL-15. Role of IL-15 in inducing memory CD8(+) T cells was analyzed by qPCR for increase in the level of Carnitine Palmitoyl Transferase 1a (CPT1a) using cDNA synthesized from RNA of PBMCs induced with different concentrations of purified bIL-15. RESULTS: Bovine IL-15 was amplified and analyzed by agarose gel electrophoresis, which showed a specific product of ~490bp, mature sequence was amplified using full-length as a template to get a product of ~350bp. The protein was expressed, purified and analyzed by SDS-PAGE and Western blotting, which showed a specific product of 32kDa. Biological activity of purified bIL-15 fusion protein showed an increase in levels of Bcl2, STAT3 and STAT5a with 5 fold, 9 fold, and 10 fold increases as analyzed by qPCR, respectively. Role of IL-15 in inducing memory T cells showed an increase in expression level of CPT1a at 2.5 fold increase as compared to control cells. CONCLUSION: Bovine IL-15 has been successfully cloned and expressed in our work, and the biological activity shows that the purified fusion protein is biologically active. As there is an increase in levels of CPT1a an enzyme critical for survival of memory T cells, IL-15 can be used for increase in the memory response, which can be used as an adjuvant with viral vaccines for increasing the immunity.

14.
Biologicals ; 42(3): 169-75, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24657207

RESUMO

In this study, thermostability of a Vero cell attenuated live camelpox vaccine under conventional lyophilization conditions has been evaluated. Three stabilizers were used separately for freeze-drying the vaccine and the stability of the vaccine, both in freeze-dried and reconstituted forms at different temperatures was assessed. The study revealed that the camelpox vaccine lyophilized with TAA stabilizer found superior with a shelf life of 44 months, 227 days, 22 days and 20 days at 4, 25, 37 and 45 °C, respectively followed by LS stabilizer. In terms of half-life, TAA stabilizer proved better followed by LS and BUGS stabilizers at all temperatures except at 25 °C in which LS found relatively superior. Among the four diluents viz. 1x PBS (phosphate buffered saline, pH 7.4), 0.85% NaCl, distilled water and 1 M MgSO4, PBS was a better diluent followed by 0.85% NaCl. Both the diluents maintained the infectivity titer more than the minimum effective dose (3 log10TCID50 with a maximum titre of 6.53 log10TCID50 in both the diluents) for 60 h at 37 and 45 °C. However, 1 M MgSO4 found less suitable for camelpox vaccine dilution. The study indicates that the TAA and 1× PBS are the choice of stabilizer and diluent, respectively for camelpox vaccine.


Assuntos
Orthopoxvirus/imunologia , Vacinas Virais/imunologia , Animais , Linhagem Celular , Chlorocebus aethiops , Liofilização , Meia-Vida , Células Vero
15.
J Virol Methods ; 201: 44-50, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24552953

RESUMO

A rapid and sensitive TaqMan based real-time duplex PCR (drt-PCR) assay for simultaneous detection, differentiation and quantitation of Capripoxvirus (CaPV) and Orf virus (ORFV) DNA, was optimized targeting the highly conserved DNA polymerase genes of these virus genomes. Two pairs of oligonucleotide primers and two hybridization probes labeled with Cy5/BHQ1 and Hex/BHQ1 for CaPV and ORFV, respectively, were used in the drt-PCR assay. The assay was found to be specific only to targeted viruses and did not react with buffalopox virus (BPXV), camelpox virus (CMLV) (Orthopoxviruses) and cDNA of Peste des petits ruminants virus and bluetongue virus, the other common viruses of sheep and goats. The detection limit of the assay was 20 copies for each of the standard plasmid and 35fg of viral genomic DNA for CaPV and ORFV, respectively, in a single and mixed virus population. Both intra-(0.49-4.6% and 0.7-3.7%) and inter-(0.6-2.35% and 0.27-2.1%) assay variations of drt-PCR for CaPV and ORFV DNA were within the acceptable limits, implying high reproducibility and repeatability of the assay. Further, the diagnostic specificity and the sensitivity of the assay was assessed using known virus isolates of sheeppox virus (SPPV), goatpox virus (GTPV) and ORFV and the clinical specimens from sheep and goats. The developed drt-PCR assay was able to detect, differentiate, quantify simultaneously and also to identity mixed infections of CaPV and ORFV in sheep and goats.


Assuntos
Capripoxvirus/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Vírus do Orf/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Medicina Veterinária/métodos , Animais , Capripoxvirus/genética , Primers do DNA/genética , DNA Polimerase Dirigida por DNA/genética , Ectima Contagioso/diagnóstico , Ectima Contagioso/virologia , Cabras , Sondas de Oligonucleotídeos/genética , Vírus do Orf/genética , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/veterinária , Infecções por Poxviridae/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Ovinos , Proteínas Virais/genética
16.
Virusdisease ; 25(1): 85-90, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24426314

RESUMO

The present study describes the prevalence of Peste-des-petits-ruminant virus (PPRV) antibodies in cattle, buffaloes, sheep and goats carried out during the period 2011 using the serum samples randomly collected from different villages of five states of India. A total of 1,498 serum samples [n = 605 (cattle); n = 432 (buffaloes); n = 173 (sheep); n = 288 (goats)] were collected from 52 districts in five states (Andhra Pradesh, Gujarat, Jammu and Kashmir, Maharashtra and Rajasthan) of India and were screened for PPRV-specific antibodies by using PPR monoclonal antibody-based competitive ELISA kit. Analysis of 1,498 samples indicates that an overall seroprevalence of 21.83 % with 11.07 % in cattle, 16.20 % in buffaloes, 45.66 % in sheep and 38.54 % in goats. This report presents the results of PPRV-specific antibodies in situations where the subclinical, inapparent or nonlethal or recovery of infection was suspected in cattle, buffaloes, sheep and goats. The presence of PPRV antibodies demonstrate that bovines are exposed to PPRV infection and it implies the importance of cattle and buffaloes as subclinical hosts for the virus besides widespread presence of the disease in sheep and goats. Further, the study showed that the prevalence of PPRV antibodies in apparently healthy livestock under natural situation, 21.83 % of the animals were protected from PPRV re-infection. This inturn help in the implementation of disease control strategies such as vaccination in that particular geographical area.

17.
J Virol Methods ; 195: 1-8, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24134940

RESUMO

A multiplex polymerase chain reaction (mPCR) was developed and evaluated for detection of pox viral infections simultaneously using clinical samples from sheep and goats. Specific primers for three pox viruses of sheep and goats including sheeppox virus (SPPV), goatpox virus (GTPV) and orf virus (ORFV) were designed targeting conserved sequences of the DNA binding phosphoprotein (I3L) coding gene of Capripoxvirus (CaPV) and the DNA polymerase (E9L) gene of parapoxvirus for identification of these viruses. The mPCR assay was found to be sensitive for detecting as low as 350 pg of viral genomic DNA or 10(2) copies of standard plasmid of individual targets; and 10(3) copies of plasmid in a mixture of two or three viruses. The assay was specific for detecting one or more of the viruses in various combinations from clinical specimens. Two hundred and thirty five (n=235) clinical samples from sheep and goats received from different geographical regions of the country for diagnosis of pox infection were evaluated by developed uniplex and mPCR assays. The assay had improved diagnostic sensitivity and specificity over to in-use laboratory diagnostic methods and can be useful for clinical differential diagnosis of these infections in sheep and goats.


Assuntos
Capripoxvirus/isolamento & purificação , Ectima Contagioso/diagnóstico , Doenças das Cabras/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Vírus do Orf/isolamento & purificação , Infecções por Poxviridae/veterinária , Doenças dos Ovinos/diagnóstico , Animais , Capripoxvirus/classificação , Capripoxvirus/genética , Primers do DNA/genética , Ectima Contagioso/virologia , Doenças das Cabras/virologia , Cabras , Técnicas de Diagnóstico Molecular/métodos , Vírus do Orf/classificação , Vírus do Orf/genética , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/virologia , Sensibilidade e Especificidade , Ovinos , Doenças dos Ovinos/virologia , Medicina Veterinária/métodos , Proteínas Virais/genética
18.
Virusdisease ; 25(3): 350-7, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25674603

RESUMO

Virulent isolate of peste des petits ruminants virus (PPRV) of Indian origin (PPRV Jhansi 2003) initially adapted in Vero cells was further propagated in thermo-adapted (Ta) Vero cells grown at 40 °C for attaining thermo-adaption and attenuation of virus for development of Ta vaccine against PPR in goats and sheep. The virus was attenuated up to 50 passages in Ta Vero cells, at which, the virus was found sterile, innocuous in mice and guinea pigs and safe in seronegative goats and sheep. The developed vaccine was tested for its immunogenicity in goats and sheep by subcutaneous inoculation of 100 TCID50 (0.1 field dose), 10(3) TCID50 (one field dose) and 10(5) TCID50 (100 field doses) of the attenuated virus along with controls as per OIE described protocols for PPR vaccine testing and were assessed for PPRV-specific antibodies 7-28 days post vaccination (dpv) by PPR competitive ELISA and serum neutralization tests. The PPRV antibodies were detected in all immunized goats and sheep and goats were protective when challenged with virulent PPRV at 28th dpv along with controls for potency testing of the vaccine. The attenuated vaccine did not induce any adverse reaction at high dose (10(5) TCID50) in goats and sheep and provided complete protection even at low dose (10(2) TCID50) in goats when challenged with virulent virus. There was no shedding and horizontal transmission of the attenuated virus to in-contact controls. The results indicate that the developed PPR Ta attenuated virus is innocuous, safe, immunogenic and potent or efficacious vaccine candidate alternative to the existing vaccines for the protection of goats and sheep against PPR in the tropical countries like India.

19.
Virusdisease ; 25(3): 408-11, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25674614

RESUMO

The present study was undertaken to investigate the possible involvement of cattle in the epidemiology of peste des petits ruminants (PPR) as subclinical carriers. Cattle were exposed experimentally to PPR virus (PPRV) infection or placed in contact with PPR infected goats. Clinical samples including heparinized/EDTA blood, plasma, peripheral blood monocyte cells (PBMCs) and clotted blood (for serum) were collected periodically from 21 days post infection (dpi) to 397 dpi (21, 45, 50, 57, 65, 95, 111, 119, 148, 190, 203 and 397 dpi) and tested for PPRV antigen, nucleic acid and antibody. Exposed cattle seroconverted and maintained PPRV specific haemagglutinin antibodies and detectable PPRV antigen/nucleic acid in blood, plasma and PBMCs from 21 to 397 dpi. PPRV was recovered from blood and PBMC collected from experimental animals at 21 dpi, initially in B95a cells and then adapted to Vero cells. The study indicated that PPRV can infect cattle subclinically and PPRV antigen/nucleic acid persist in cattle for at least 397 days.

20.
Indian J Virol ; 24(3): 373-9, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24426300

RESUMO

Peste des petitis ruminants (PPR) is an economically important endemic viral disease of sheep and goats in India, where several different homologous PPR vaccine candidates have been developed. We evaluated the serological response to two vaccine strains, Arasur/87 and Sungri/96, in South Indian cross-bred and native sheep and goats reared under organized and unorganized settings. Animals seronegative (percent inhibition or PI <40) by competitive enzyme-linked immunosorbent assay (c-ELISA) were immunized with either of the vaccine strains or placebo. Sera collected on 21, 60 and 90 days post-vaccination were subjected to c-ELISA and serum neutralization test (SNT). Seropositivity (PI >40), seroconversion (fourfold increase in SNT titres) and seroprotection (SNT titre of ≥8 deemed to be protective) ranged from 66.7 to 84.0 %, 56.0 to 69.2 %, and 60.0 to 76.0 %, respectively. However, no significant difference was observed between responses to the two vaccine strains. These results support the premise that the two vaccine strains are equally efficacious.

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