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The present study examined 434 field samples including serum (n = 273), swabs from natural orifices (n = 52) and postmortem tissue samples (n = 109) from both suspected and asymptomatic swine from Andhra Pradesh, Karnataka, Kerala, Pondicherry, Tamil Nadu, and Telangana states in southern India. All the samples were processed for molecular screening of PCV3 by specific PCR assay. Overall molecular positivity rate of PCV3 was found to be 0.7% in southern India with one sample positive from each state of Tamil Nadu, Kerala and Telangana. All the three PCR positive PCV3 samples are detected from reproductive failures and were processed and propagated in PK15 cell line for virus isolation. Out of 3 samples processed, one (INDKL9PK76) PCV3 isolate could be obtained in this study and it was confirmed by specific PCR at third and fifth passage levels. Sequencing of PCV3 positive PCR amplicon (INDKL9PK76) revealed 1004 nucleotides and BLAST analysis confirmed partial sequence of the PCV3 genome. The aligned contig sequence was submitted to GenBank under the accession number of MW627201. PCV3 sequence in this study revealed 99% homology with PCV3 isolates from Europe and China. Phylogentic analysis of the PCV3 isolate-INDKL9PK76 sequence along with established PCV3 genotypes revealed clustering within PCV3 genotypes. Characterization of PCV3 (INDKL9PK76) isolate based on deduced amino acid composition of PCV3-capsid protein revealed "A" (alanine) and "R" (arginine) at 24th and 27th residues respectively confirming the incidence of PCV3a genotype. This study evidences PCV3 associated reproductive failure in domestic pigs for the first time in southern India.

The objective of this experiment was to study and compare the effects of dietary supplementation of organic and inorganic zinc (Zn) on growth performance, nutrient utilisation and gene expression pattern of glucose transporter protein in peripheral blood mononuclear cells (PBMC) in Malabari kids. Fifteen, 3–4-month-old goat kids were divided into three groups uniformly by using completely randomised design (CRD). Group G1 was fed on basal diet as per NRC requirement, and G2 and G3 were fed on basal diet + 40 ppm Zn as inorganic zinc sulphate (ZnSO4) and 40 ppm Zn as organic Zn methionine, respectively, for a period of 91 days. Supplementation of inorganic and organic Zn had no significant effect on dry matter (DM) intake. The digestibility of crude protein (CP), ether extract (EE), neutral detergent fibre (NDF), hemicellulose and cellulose was significantly more in the organic Zn–supplemented group. The average daily gain and feed:gain ratio were significantly (p

This study used 56 aborted and stillborn fetuses from organized swine farms in Tamil Nadu and Kerala, southern states of India. All samples were screened by using a PCR assay that targets the NS1 gene for PPV. Furthermore, the PCR positive samples were subjected to amplification of the VP2 gene of PPV1 with designed primers and sequenced for further study. The PCR screening of 56 samples found that 14.3% (n = 8) were positive for PPV genome. According to VP2 gene-based PCR for PPV1, 897 bp specific amplicons were detected in all eight of the samples. Two of the eight positive samples (L17 and T5) were sequenced and annotated randomly. The BLAST analysis of contig sequence INDTNCHN-T5 revealed 100% sequence homology with Chinese PPV1genome, whereas sequence from INDTNCHN-L17 revealed 99.43% sequence homology with Spain, Chinese, and German. PPV1 sequences and both the sequences INDTNCHN-T5 and INDTNCHN-L17 were submitted to the GenBank under the accession numbers MW822566 and MW822567 respectively. A phylogenetic analysis of the sequences in this study revealed specific grouping along with PPV1 strains in cluster E. Amino acid analysis of both isolated sequences in addition to the reference sequence from PPV1 showed variations in position 215 (I to T) in both the isolates, variation at position 228 (Q to E) in T5 isolate and variations at position 59 (L to M) and 314 (K to E) in L17 isolate. This study represents the first report of PPV1 cluster E in Tamil Nadu, southern India.

Newcastle disease virus (NDV) can infect approximately 250 avian species and causes highly contagious Newcastle disease (ND) in domestic poultry, leading to huge economic losses. There are three different pathotypes of NDV, i.e., lentogenic, mesogenic, and velogenic. Wild resident (wild) and migratory birds are natural reservoirs of NDV and are believed to play a key role in transmitting the virus to domestic poultry. The present study was conducted to determine the prevalence of NDV in wild and migratory birds in the state of Haryana, India, during two migratory seasons (2018-19 and 2019-20). In total 1379 samples (1368 choanal swabs and 11 tissue samples) were collected from live (in/i= 1368) or dead birds (in/i= 4) belonging to 53 different avian species. These samples belonged to apparently healthy (in/i= 1338), sick (in/i= 30), and dead (in/i= 4) birds. All samples were tested for NDV by real-time reverse transcription-PCR using M gene specific primers and probe. Of the 1379 samples, 23 samples from wild birds [iColumba livia domestica/i(in/i= 12, 52.17%),iPavo cristatus/i(in/i= 9, 39.13%), andiPsittaciformes/i(in/i= 2, 8.69%)] were found positive for NDV. Only one of the 23 samples (fromiP. cristatus/i) was positive for F gene, indicating it to be a mesogenic/velogenic strain. These results indicate that both lentogenic and velogenic strains of NDV are circulating in wild birds in Haryana and that further studies are needed to characterize NDV strains from wild/migratory birds and domestic poultry to determine the extent of virus transmission among these populations. This study considers the disease transmission risk from domestic pigeons and parrots to commercial poultry and vice versa, and the results emphasize the need for strict biosecurity strategies to protect commercial poultry in the region.Prevalencia del virus de la enfermedad de Newcastle en aves silvestres y migratorias en Haryana, India. El virus de la enfermedad de Newcastle (NDV) puede infectar aproximadamente a 250 especies de aves y causa la enfermedad de Newcastle (ND) altamente contagiosa en la avicultura comercial, lo que genera enormes pérdidas económicas. Hay tres patotipos diferentes del virus de Newcastle, que incluyen, lentogénico, mesogénico y velogénico. Las aves silvestres residentes (silvestres) y migratorias son reservorios naturales del virus de Newcastle y se cree que desempeñan un papel clave en la transmisión del virus a las aves domésticas comerciales. El presente estudio se realizó para determinar la prevalencia del virus de Newcastle en aves silvestres y migratorias en el estado de Haryana, India, durante dos temporadas migratorias (2018-19 y 2019-20). En total, se recolectaron 1379 muestras (1368 hisopos coanales y 11 muestras de tejido) de aves vivas (in/i= 1368) o muertas (in/i= 4) pertenecientes a 53 especies de aves diferentes. Estas muestras pertenecían a aves aparentemente sanas (in/i= 1338), enfermas (in/i= 30) y muertas (in/i= 4). Todas las muestras se analizaron para detectar al virus de Newcastle mediante transcripción reversa y PCR en tiempo real utilizando iniciadores y una sonda específicos del gene M. De las 1379 muestras, 23 muestras de aves silvestres [iColumba livia domestica/i(in/i= 12, 52.17 %),iPavo cristatus/i(in/i= 9, 39.13 %) yiPsittaciformes/i(in/i= 2, 8.69 %)] resultaron positivas para el virus de Newcastle. Solo una de las 23 muestras (deiP. cristatus/i) fue positiva para el gene F, lo que indica que se trata de una cepa mesogénica/velogénica. Estos resultados indican que tanto las cepas lentogénicas como las velogénicas del virus de Newcastle están circulando en las aves silvestres de Haryana y que se necesitan más estudios para caracterizar las cepas del virus de Newcastle de las aves silvestres/migratorias y de las aves domésticas para determinar el alcance de la transmisión del virus entre estas poblaciones. Este estudio considera el riesgo de transmisión de la enfermedad de las palomasdomésticas y loros a las aves comerciales y viceversa, y los resultados enfatizan la necesidad de estrategias estrictas de bioseguridad para proteger las aves comerciales en la región.

Infectious bursal disease (IBD) is considered as menace as it affects poultry industry globally causing immunosuppression, high mortality and heavy economic loss. Outbreaks of IBD were reported in many states of India including Kerala. VP1 gene acts as an important factor in the process of virus encapsidation and its involvement in viral virulence and viral replication indicates its importance in infectious bursal disease virus (IBDV). The present study was conducted to carry out the molecular characterization of VP1 gene of virulent IBDV in Kerala. A total of 42 samples were processed for the detection and analysis of VP1 gene of IBDV. Out of 42 samples, 21 samples were positive for VP1 gene of IBD. The phylogenetic analysis of the partial VP1 gene sequences reveals the clustering of IBDV isolates into very virulent IBDV (vvIBDV) and non-virulent IBDV (vIBDV). Eighteen isolates (11 isolates from vaccinated flock and 7 from non-vaccinated flocks) clustered with very virulent strains. Three isolates (2 isolates were from vaccinated flock and 1 from non-vaccinated flock) clustered with non-virulent IBDV strains, showing more evolutionarily similarity to south Indian strain VCN14/ABT/MVC/India. It is observed that vvIBDV isolates from this study have common ancestor with the south Indian strain PY12 but showed 9–10% divergence from this strains. The amino acid analysis of these 21 isolates revealed that 17 isolates possessed the characteristic vvIBDV TDN amino acid triplet, while the three isolates had non-vIBDV NEG amino acid triplet at 145/146/147 position. The remaining isolate 1/CVASP/IBDV/VP1 shows unique PDN triplet instead of TDN. Two vvIBDV isolates (15/CVASP/IBDV/VP1 and 18/CVASP/IBDV/VP1) showed 100% nucleotide and amino acid similarity with intermediate plus vaccine strain. Four vvIBDV isolates showed neutral amino acid substitution K251R which was earlier reported in Indian strains but first time in south Indian isolates. The most common unique amino acid substitution observed in our study was neutral E269D amino acid substitution in 12 isolates, neutral amino acid substitution T329S in five isolates, neutral T174N and non-polar to polar amino acid substitution A178T in isolate 10/CVASP/IBDV/VP1, non-polar to polar amino acid substitution P360R in isolate 17/CVASP/IBDV/VP1 and non-polar to polar amino acid substitution P188S in isolate 1/CVASP/IBDV/VP1. These novel mutations in our study reveal the role of genetic drift in the evolution of vvIBDV strains. The isolate 2/CVASP/IBDV/VP1 from non-vaccinated flock shows VP1 gene of non-vIBDV, but possessing VP2 of vvIBDV type indicates this is evolved by genetic shift of segments A and B. This is the first genetic characterization study of field VP1 gene of IBDV isolates in Kerala, India.

Bluetongue virus (BTV) is the ‘type’ species of the genus Orbivirus within the family Reoviridae. The BTV genome is composed of ten linear segments of double-stranded RNA (dsRNA), each of which codes for at least one of ten distinct viral proteins. Phylogenetic analysis based on VP2, VP3, VP5, VP7 and NS3 gene has been advocated by different researchers around the world. However, very little information about the phylogenetic analysis based on the NS1 gene of BTV-16 isolates is available. In this study, a partial sequence of segment 5 (Seg 5) from the BTV-16 Hisar isolate was sequenced. Sequence analysis of the Seg 5 of the BTV-16 Hisar isolate revealed highest nucleotide sequence identity of 98.9% (BTV-21), -96.4% (BTV-9) with sequences of NS1 genes of Indian isolates and 97.8% (BTV-3),-93.4%(BTV-1) isolates from other countries. The lowest sequence identity detected was 91.2% between sequences of NS1 gene of JQ924824 of BTV 16 Indian isolate and JX861502 BTV-1 of France. The study also assessed the comparative identity from the deduced amino acid sequences and found maximum homology (100% identity) with Indian isolates and 95.2 to 98.8% identity with Western isolates with only three amino acid difference. Phylogenetic analysis of the NS1 gene of the BTV-16 Hisar isolate with others around the world has revealed two major clusters which includes the Indian (Eastern) linage and Western linage separately. The BTV-16 Hisar isolate was found to be closely related with the BTV-9 and 21 Indian isolates based on its NS1 gene. Phylogenetic analysis of segment 5 is highly conserved among the different serotypes however, NS1 of BTV-3 of USA clustered with the Eastern lineage indicating introduction of Western BTV strains and a possible reassortment between Eastern and Western field strains in India. Key words: Bluetongue virus, genome, NS1, Orbivirus, phylogeneticanalysis, segment-5, serotype.

Ticks and tick-borne diseases (TTBDs) are considered major causes of economic loss in the livestock sector which incur an annual control cost estimated at US$ 498.7 million in India. Among these diseases, babesiosis, theileriosis and anaplasmosis are listed among the top ten livestock diseases in India and cause significant mortality and morbidity among cattle. However, molecular characterization of bovine Babesia and Anaplasma species are scant; thus, the aim of this study is to perform molecular characterization of field isolates of Babesia spp. and Anaplasma spp. infecting bovines in Kerala, South India. Blood smears and whole blood samples were collected from a total of 199 apparently healthy adult female cattle in Kerala. Based on microscopy, Babesia spp., Theileria orientalis and Anaplasma spp. organisms were detected in 9 (4.5%), 40 (20%) and 6 (3%) samples, respectively. Genus-specific polymerase chain reactions for amplification of 18S rRNA of Babesia spp. and 16S rRNA of Anaplasma spp. revealed positive results with 18 (9%) and 14 (7%) samples. The phylogenetic analysis of 18S rRNA gene sequences of Babesia spp. confirmed the existence of two different populations of Babesia spp. circulating in the blood of infected cattle viz., Babesia bigemina and a Babesia sp. genetically related to Babesia ovata. Further phylogenetic analysis using rap-1a sequences of isolates of B. bigemina revealed higher levels of genetic heterogeneity. However, the field isolates of B. bigemina displayed only slight heterogeneity when the rap-1c gene was examined. Polymerase chain reaction followed by sequencing and phylogenetic analysis of 16S rRNA gene of Anaplasma spp. revealed the existence of Anaplasma marginale, Anaplasma bovis and Anaplasma platys in bovines in South India. Based on msp4 gene sequences, all the field isolates of A. marginale from Kerala were clustered in a single clade with others isolated from around the world. To our knowledge, this study forms the first report on occurrence of B. ovata-like parasites and A. platys in cattle from India.

Infectious bursal disease (IBD) is considered as menace as it affects poultry industry globally causing immunosuppression, high mortality and heavy economic loss. Outbreaks of IBD were reported in many states of India including Kerala. VP1 gene acts as an important factor in the process of virus encapsidation and its involvement in viral virulence and viral replication indicates its importance in infectious bursal disease virus (IBDV). The present study was conducted to carry out the molecular characterization of VP1 gene of virulent IBDV in Kerala. A total of 42 samples were processed for the detection and analysis of VP1 gene of IBDV. Out of 42 samples, 21 samples were positive for VP1 gene of IBD. The phylogenetic analysis of the partial VP1 gene sequences reveals the clustering of IBDV isolates into very virulent IBDV (vvIBDV) and non-virulent IBDV (vIBDV). Eighteen isolates (11 isolates from vaccinated flock and 7 from non-vaccinated flocks) clustered with very virulent strains. Three isolates (2 isolates were from vaccinated flock and 1 from non-vaccinated flock) clustered with non-virulent IBDV strains, showing more evolutionarily similarity to south Indian strain VCN14/ABT/MVC/India. It is observed that vvIBDV isolates from this study have common ancestor with the south Indian strain PY12 but showed 9-10% divergence from this strains. The amino acid analysis of these 21 isolates revealed that 17 isolates possessed the characteristic vvIBDV TDN amino acid triplet, while the three isolates had non-vIBDV NEG amino acid triplet at 145/146/147 position. The remaining isolate 1/CVASP/IBDV/VP1 shows unique PDN triplet instead of TDN. Two vvIBDV isolates (15/CVASP/IBDV/VP1 and 18/CVASP/IBDV/VP1) showed 100% nucleotide and amino acid similarity with intermediate plus vaccine strain. Four vvIBDV isolates showed neutral amino acid substitution K251R which was earlier reported in Indian strains but first time in south Indian isolates. The most common unique amino acid substitution observed in our study was neutral E269D amino acid substitution in 12 isolates, neutral amino acid substitution T329S in five isolates, neutral T174N and non-polar to polar amino acid substitution A178T in isolate 10/CVASP/IBDV/VP1, non-polar to polar amino acid substitution P360R in isolate 17/CVASP/IBDV/VP1 and non-polar to polar amino acid substitution P188S in isolate 1/CVASP/IBDV/VP1. These novel mutations in our study reveal the role of genetic drift in the evolution of vvIBDV strains. The isolate 2/CVASP/IBDV/VP1 from non-vaccinated flock shows VP1 gene of non-vIBDV, but possessing VP2 of vvIBDV type indicates this is evolved by genetic shift of segments A and B. This is the first genetic characterization study of field VP1 gene of IBDV isolates in Kerala, India.

Group A rotaviruses (GAR) are an important cause of diarrhoea in infants and newborn animals especially pigs. In this paper, we report the detection, G and P typing and phylogenetic analysis of GAR of pigs in Kerala. A total of 100 fecal samples from diarrhoeic piglets were collected from organized farms in Wayanad, Ernakulam, Thrissur, and Palakkad districts of Kerala. The samples were tested for the presence of GAR employing reverse transcriptase polymerase chain reaction (RT-PCR) targeting VP6 gene. Positive samples were tested by G and P genotyping primers and representative amplicons were sequenced. Of the 100 samples, 12 were positive for GAR. The G and P types detected were G2, G4, G5, G6, G9, P[6] and P[19]. An untypable P type (P21-5 like) was also detected. In some of the samples more than one G type was detected. The nucleotide sequences of G2, G4 and G5 types were similar to those seen in pigs and that of G6 was similar to bovine sequences. G9, P[6] and P[19] sequences showed similarity to human rotavirus sequences. The findings of this study provide the first information on the G and P genotypes of GAR of pigs in Kerala. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13337-020-00621-y) contains supplementary material, which is available to authorized users.

Porcine reproductive and respiratory syndrome (PRRS) caused by an arterivirus is characterised by reproductive disorders in sows, and post-weaning pneumonia and growth reduction in piglets. Though the virus has been detected in Kerala, no systematic study has been carried out to ascertain its genotype and molecular epidemiology. In the present study, 7 PRRS virus (PRRSV) positive samples collected from incidences of PRRS in Kerala during 2017–2019 were subjected to ORF5, ORF7 and Nsp2 gene based reverse transcription polymerase chain reaction and the specific amplicons generated were sequenced. On BLAST analysis it was revealed that all the sequences were of genotype 2 (North American genotype). Phylogenetic analysis of ORF5 sequences, grouped them under subgenotype 4 with close clustering with other isolates from Kerala, Mizoram and Assam. Nsp2 gene sequence based phylogenetic analysis grouped the isolates under subgenotype 3 with similarities to isolates from Mizoram. Phylogenetic analysis based on ORF7, clustered the isolates under study with PRRSV isolates from Mizoram and Meghalaya. In Nsp2 sequences, a 30 amino acid discontinuous deletion was observed. On analysis of amino acid sequences of ORF5 of Kerala isolates and those from India, it was seen that the Kerala isolates showed closer similarity to PRRSV isolates from Assam than to the other Indian isolates. The study reveals that PRRSV strains prevalent in Kerala share close relationship with other PRRSV isolates in India. This may be due to spread of the virus from these regions to Kerala due to animal movement. Concerted efforts should be undertaken to check unauthorized animal movement to control spread of this economically important disease. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13337-020-00634-7) contains supplementary material, which is available to authorized users.

The molecular characterization of three Newcastle disease viruses (NDV) isolated from backyard chickens in the state of Haryana, India, was undertaken. Two genotype II strains and one genotype XIIIc class II isolate with genome sizes of 15,186 and 15,192 nucleotides (nt), respectively, were identified.

Recurrent infectious bursal disease (IBD) outbreaks were reported in different regions of Kerala, India. This paper reports the comparative genetic analysis of the hypervariable region of the VP2 gene of IBD virus isolates from the field outbreaks in Kerala. In phylogenetic analysis, the obtained field isolates fall into genogroup 1 and 3. In genogroup 3, all vvIBDV isolates shared a common ancestor with other south Indian isolates but isolates 9/CVASP/IBDV, 10/CVASP/IBDV, 12/CVASP/IBDV, 14/CVASP/IBDV and 17/CVASP/IBDV are most recently evolved and are diverged from the south Indian isolates. The amino acid sequence of 22 isolates was analysed, out of which 18 had conserved amino acids which were characteristic of vvIBDV. All the vvIBDV isolates obtained in the study had phenylalanine and valine at the position 240 and 294, respectively, similar to recently evolved Indian IBDV isolate (MDI14). But we observed T269A and S299N mutations in the isolate 6/CVASP/IBDV, and it is the first report of such mutations at these positions in India IBDV isolates. The isolate 11/CVASP/IBDV had a unique mutation of V225A which is not yet reported in IBDV isolates. Two isolates (15/CVASP/IBDV and 18/CVASP/IBDV) were 100% amino acid similar to intermediate plus vaccine strain. The isolates 8/CVASP/IBDV/VP2 and 19/CVASP/IBDV had amino acids unique for the intermediate vaccine with mutations observed at H253Q and V256I in 19/CVASP/IBDV, T270A and novel mutation N279Y in isolate 8/CVASP/IBDV. These two isolates had non-virulent classical heptapeptide sequence 'SWSARGS'; nevertheless, they produce field outbreaks of IBD. This is the first report of genetic characterisation of IBDV in Kerala, India.

Classical swine fever (CSF) is an economically important disease of pigs caused by CSF virus (CSFV) belonging to the genus Pestivirus within the family Flaviviridae. The disease is endemic in many countries including India. A comprehensive study was carried out to assess the type of CSFV circulating in the South Indian state of Kerala. During the period 2013–2014, clinical samples were collected from 19 suspected CSF outbreaks of domestic pigs in different districts of Kerala. The samples were tested using nested reverse transcription PCR (RT-PCR) targeting the E2 gene and RT-PCR for 5′UTR of the virus. Partial 5′ UTR and E2 gene regions of six CSFV isolates were sequenced. Phylogenetic analysis revealed that all the CSFV isolates belonged to subgroup 2.2. The isolates showed close resemblance to the other CSFV isolates circulating in India. It was also observed that the CSFV viruses from Kannur district were distinct from those circulating in the other districts as evidenced by their divergence from other Kerala isolates in the phylogenetic tree. Close relationship was seen to the CSFV isolates from South East Asian countries.

Classical swine fever (CSF) is one of the most devastating epizootic diseases of pigs, causing high morbidity and mortality worldwide. The diversity of clinical signs and similarity in disease manifestations to other diseases make CSF difficult to diagnose with certainty. The disease is further complicated by the presence of a number of different strains belonging to three phylogenetic groups. Advanced diagnostic techniques allow detection of antigens or antibodies in clinical samples, leading to implementation of proper and effective control programs. Polymerase chain reaction (PCR)-based methods, including portable real-time PCR, provide diagnosis in a few hours with precision and accuracy, even at the point of care. The disease is controlled by following a stamping out policy in countries where vaccination is not practiced, whereas immunization with live attenuated vaccines containing the ‘C’ strain is effectively used to control the disease in endemic countries. To overcome the problem of differentiation of infected from vaccinated animals, different types of marker vaccines, with variable degrees of efficacy, along with companion diagnostic assays have been developed and may be useful in controlling and even eradicating the disease in the foreseeable future. The present review aims to provide an overview and status of CSF as a whole with special reference to swine husbandry in India.

Porcine parvovirus (PPV) is a DNA virus of the genus Parvovirus of family Parvoviridae. It is the causative agent of many disease problems in pigs such as maternal reproductive failure, stillbirth, mummification, embryonic or fetal death, infertility, abortion and neonatal death. A study was conducted to assess the incidence of the virus in pigs in Kerala State in South India. A total of 38 samples were collected from domestic and wild pigs from different districts of the State. Polymerase chain reaction targeting a 265 bp fragment of the NS1 gene of the virus was carried out. Of the samples tested, 2 (5.26 %) were found to be positive for PPV virus genome, one of which was from a wild pig. One of the positive samples was sequenced and the nucleotide sequence obtained was compared with other sequences of PPV from India and abroad. The results revealed that the sequence had very close similarity to PPV sequences previously reported from India and to that of Chinese isolates. This is the first report of the existence of PPV in domestic and wild pigs in Kerala, India. The study highlights the need to test for the presence of PPV in addition to other infectious agents in diagnosis of cases of reproductive disorders in pigs.

Bovine herpesvirus -1 (BHV-1) is the etiological agent of many clinical syndromes in cattle which causes huge economic losses to the animal husbandry sector annually. Since the first report of its presence in India in 1976, the disease is considered to be endemic in the country. In the present study, a case of keratoconjunctivitis in a cow was investigated to find out the underlying cause of the condition. The clinical material (ocular swab) was tested by BHV-1 glycoprotein D gene specific PCR using in house designed primers and found to be positive by the presence of a 212 bp DNA product in agarose gel electrophoresis. The virus was isolated in MDBK cell line in the third passage and the serum from the animal, was positive for antibodies against BHV-1 by ELISA. A 575 bp segment of the glycoprotein C gene of the isolate was amplified by PCR, cloned and sequenced. On phylogenetic analysis, it was seen that the sequence matched with published BHV-1.1 sequences from USA and Uruguay whereas it was divergent from Brazilian BHV-1.1 isolates. This study highlights the isolation, rapid and sensitive detection of BHV-1 virus from clinical cases and its subtyping by nucleotide sequencing and subsequent phylogenetic analysis which gives invaluable information about the molecular epidemiology of BHV-1 subtypes prevalent in the country.

MarekÂ’s disease (MD) is a highly contagious oncogenic and neuropathic disease of chickens responsible for great economic losses to the poultry industry worldwide. An investigation was done to assess the status of MD in Kerala by serological tests viz., agar gel immunodiffusion (AGID) test, enzyme linked immunosorbant assay (ELISA) and indirect fluorescent antibody (IFA) test. Among the three tests employed for detection of antibodies to MD virus (MDV), out of 1030 sera screened, 157 samples (15.2 per cent) were positive by ELISA, whereas 62 samples were positive by AGID, which represents 6 per cent. The IFA tests detected 95 positive samples from 1030 samples (9.2 per cent). The results were analysed statistically using CochranÂ’s Q and FischerÂ’s exact test and it was found that ELISA was more sensitive than the other two tests compared.

The results presented here record the first confirmation of bluetongue virus (BTV) antibody in sheep and goats in Kerala State. A total of 1,010 sera collected from the 14 districts within the state were screened for the presence of group-specific BTV antibodies by dot enzyme-linked immunosorbent assay (dot ELISA). Positive samples were obtained from 12 of the 14 districts. The overall BTV antibody prevalence was 5.1 +/- 1.9% (at 95% confidence level) although the prevalence levels were consistently higher in organised farms than in the field population. Comparative tests carried out using the dot ELISA and competitive ELISA (C ELISA) showed a good agreement for all the positive sera. The dot ELISA was simple to perform, economic and rapid, and is therefore ideally suited for routine screening for BTV antibody at the farm level.

Rotaviral enteritis has emerged as a major cause of morbidity and mortality in piglets during their post-natal life. The present study was carried out to examine high-mobility group box 1 (HMGB1) protein, acute phase response and oxidative stress indices in the serum of suckling piglets suffering from enteritis with or without association of porcine group A rotavirus infection. The present investigation utilized 23 clinical cases with signs of acute enteritis and 12 more healthy piglets of a similar age group as control animals. Out of 23 enteritis cases, 12 cases were found to be positive for porcine group A rotavirus infection as confirmed by reverse transcription-polymerase chain reaction (RT-PCR) using specific primers for group A rotavirus, and the rest were found negative. The acute enteritis cases in piglets were associated with an elevated level of HMGB1 protein and serum haptoglobin and ceruloplasmin suggestive of an acute phase response. Among the oxidative stress indices, the concentrations of malondialdehyde (MDA) and nitric oxide (NO) in serum were significantly increased. A pronounced drop of total antioxidant capacity and the activity of antioxidant enzymes such as catalase and superoxide dismutase in the serum of piglets suffering from acute enteritis compared to healthy ones were also noticed. The alterations in HMGB1 protein, acute phase response and oxidative stress indices were more pronounced in cases with the involvement of porcine rotavirus as compared to rotavirus-negative cases. It is concluded that HMGB1 protein, markers of oxidative stress and acute phase proteins might play an important role in the aetiopathogenesis of porcine diarrhoea caused by rotavirus and might be true markers in diagnosing the conditions leading to the extension of the prompt and effective therapeutic care.

In this study, sera from 65 breeding and 19 training bulls from Uttar Pradesh State in north India were tested for bovine herpesvirus 1 (BHV-1) antibodies by enzyme linked immunosorbent assay (ELISA) and virus neutralization test (VNT). The VNT test could detect 56 (86.15%) and 9 (47.37%) of the samples from breeding and training bulls as positive for BHV-1 antibodies whereas in ELISA 63 (96.92%) and 10 (52.63%) were found positive, respectively. Semen samples from the breeding bulls were simultaneously tested by the Taqman based real time PCR (qPCR). Of the 65 samples screened, only 40 (61.54%) were found to contain BHV-1 DNA indicating that all the seropositive bulls are not shedding the virus in semen. When the RT-PCR positive samples were subjected to virus isolation on Madin-Darby bovine kidney (MDBK) cells, no virus isolates could be obtained. The advantages of concomitant testing of serum and semen of breeding bulls and measures for control of BHV-1 infections in bull farms are discussed.

The disease condition attributed to have been caused by Theileria orientalis is generally benign. However, it is also thought that the parasite, at least some strains of it, can cause fatal disease. The present communication deals with the clinical signs, postmortem lesions and diagnosis of a fatal disease due to T. orientalis which caused mortality in crossbred adult bovines of South India. High body temperature, lacrimation, nasal discharge, swollen lymph nodes and haemoglobinuria were the symptoms observed. The postmortem lesions observed were punched out ulcers in abomasum, enlargement of spleen, massive pulmonary oedema, frothy exudates in trachea, epicardial and endocardial haemorrhage and haemorrhagic duodenitis. Peripheral blood smear examination revealed rod shaped Theileria sp. organisms. Polymerase chain reaction that amplify the T. orientalis specific P(32/33) gene, followed by cloning and sequencing, revealed maximum homology with Narathiwat (Thailand) and Jingole -1 (Indonesia) isolates which were positioned as isolate type 7 of T. orientalis.