Search

Foot-and-mouth disease (FMD) is endemic in India and causes severe economic loss. Status of FMD in the country for five fiscal years is presented. Outbreaks were more in number in 2007-2008 than 2010-2011. Three serotypes of FMD virus (O, A and Asia1) are prevalent. Serotype O was responsible for 80% of the confirmed outbreaks/cases, whereas Asia1 and A caused 12% and 8%, respectively. Geographical region-wise assessment indicated varying prevalence rate in different regions viz; 43% in Eastern region, 31.5% in Southern region, 11.6% in North-eastern region, 5% Central region, 4.4% Western region and 4% in Northern region. Highest number of outbreaks/cases was recorded in the month of September and lowest in June. Emergence and re-emergence of different genotypes/lineages within the serotypes were evident in real-time investigation carried out from time to time. Continues antigenic divergence in serotype A resulted in change in the vaccine strain in 2009. As on date, all genetic diversity within the serotypes is well tolerated by the vaccine strains. Unrestricted animal movements in the country play a major role in the spread of FMD. [ABSTRACT FROM AUTHOR]

Abstract: Thirteen BVDV isolates collected in four geographic regions of India between 2000 and 2002 were typed in 5′-UTR. To confirm results of genetic typing, selected viruses were also analysed in the Npro region. Phylogenetic analysis revealed that all Indian BVDV isolates belong to BVDV-1b (Osloss-like group). Despite a long distance between the farms from which the viruses were isolated there was no correlation between the origin of viral isolates and their position in a phylogenetic tree. Higher genetic similarity of Indian BVDV isolates was observed most probably due to the uncontrolled movement of cattle as well as the uncontrolled use of semen from bulls for breeding of local and farm cattle in different states of India. [Copyright &y& Elsevier]

Outbreaks of highly pathogenic avian influenza (HPAI) H5N1 virus were reported for the first time in India during February 2006. Herein, we have sequenced and analyzed the PB2 genes of five influenza virus isolates obtained from three affected states (Gujarat, Madhya Pradesh and Maharashtra) in India during the outbreaks. In the phylogenetic analysis, the Indian isolates were grouped in the mixed-migratory bird sub-lineage of the Eurasian lineage. From the phylogenetic tree, it is evident that viruses were probably introduced to India from China via Europe because they share a direct ancestral relationship with the Indian isolates. The virus might have spread through migratory waterfowls that survived the HPAI H5N1 infection. These viruses were able to replicate in cultured cells of avian and mammalian hosts and posses lysine at position 627 of the PB2 protein, indicating that they might be able to cross the host barrier to infect mammals. [ABSTRACT FROM AUTHOR]

Summary: A bidirectional axial‐flow impulse turbine installed in an oscillating water column raises the question of damping at different wave conditions. This article reports unsteady performances of such a turbine in an airflow test rig. The rig has a piston‐cylinder arrangement, and the piston, when reciprocating, generates the oscillatory airflow. At a very low wave height (piston stroke length, <0.4 m), the turbine speed is very low. Time series analysis of pressure drop and speed are reported for higher stroke lengths, which shows the rate of increase and decrease in velocity of the turbine. Furthermore, the turbine pressure coefficient and the damping characteristics of the test rig chamber were estimated. Finally, the chamber damping verifies that the turbine‐chamber coupling effect is reciprocal. [ABSTRACT FROM AUTHOR]

This study compared the therapeutic efficacy of levofloxacin, ornidazole and alpha tocopherol combination and prostaglandin F2a (PGF2a) in longstanding cases of endometritis and evaluated their impact on Interleukin-6 (IL-6) and Interleukin-10 (IL-10) transcript level in peripheral blood leukocytes. Eighteen endometritic crossbred Jersey cows were randomly allotted to three groups (six in each) viz. Group I (levofloxacin combo treatment I/U), group II (PGF2a treatment I/M), group III (no treatment, control), and group IV (six non-endometritic healthy cyclic) was taken for comparison study. The clinical efficacy was assessed by haematological study (TLC: Total leukocyte count; DC: Differential count), polymorphonuclear leukocytes (PMN) count in uterine cytology and relative mRNA expression of IL-6 and IL-10 in peripheral blood leukocytes before and after treatment with respect to conception rate following single and second inseminations. Group I and II registered significant increase in TLC and neutrophil count. PMN cytology was increased two and three fold in group I and II, respectively. The IL-6 transcript level was increased by 2.5 and 4.6 fold while that of IL-10 increased by 3.7 and 5.2 fold in group I and II, respectively. Conception rate across group I to IV following single insemination was found to be 66.67%, 50%, 16.67%, and 83.33% and their corresponding values following second insemination were 66.67%, 83.33%, 16.67%, and 83.33%, respectively. Thus, the administration of levofloxacin combo and PGF2a might have better conception rate following first and second insemination, respectively. Our study also reveals that PGF2a could register better clearance of bacteria through stronger PMN cell and cytokine activity in post-treatment period. [ABSTRACT FROM AUTHOR]

Summary: Foot‐and‐mouth disease (FMD) is, arguably, the animal disease with the most devastating global economic impact owing in part, to the severe trade restrictions imposed upon affected countries and regions. South Asia is one of the regions where widespread lineages of the FMDV virus (FMDV) have emerged. Here, we performed an integrative phylogenetic analysis of all FMDV serotypes (A, O and Asia‐1) circulating in southern Asia, including viral sequences collected until 2013. Our results describe the occurrence of FMD caused by different serotypes and lineages, focusing in the cycles where a specific lineage predominates within a region for a protracted period and then are rapidly or progressively replaced by an emergent or re‐emergent strain that is introduced from an adjacent region. Transmission between the two main regions in southern Asia (the Indian subcontinent and the region comprised by Afghanistan, Iran and Pakistan) has been limited. Results of time divergence estimation of lineages that currently circulate in this region indicate that the most recent common ancestor of endemic lineages are: 1992 [1989–1995] for lineage O/PanAsia; 1997 [1995–1999] for PanAsia2; 2001 [1998–2004] for O/Ind2001; 2001 [2000–2002] for A/Iran‐05; 1990 [1988–1991] for A/G‐18 (G‐VII); 2003 [2000–2006] for Asia‐1 Sindh08 and 2002 [1999–2004] for Asia‐1 G‐VIII. We estimated the mean of the overall substitution rate of the VP1 coding region (substitution/site/year) for serotype O (5.95 × 10−3), serotype A (1.19 × 10−2) and serotype Asia‐1 (3.08 × 10−3). The potential factors driving the lineage turnover are discussed. Our results provide insights into the ecological and evolutionary factors driving the emergence of FMDV. [ABSTRACT FROM AUTHOR]

The present paper describes the investigation of foot-and-mouth disease (FMD) outbreak in a private pig farm at Kotty in Kollam district of Kerala during October 2013. During the clinical phase, severe vesicular lesions on snout and skin around the coronary bands were observed in pigs. A total of 48 serum samples and 12 clinical samples (ruptured snout epithelia) were collected. All serum samples were subjected to indirect 3AB nonstructural protein (NSP) ELISA and liquid phase blocking (LPB) ELISA. In 3AB NSP ELISA, all serum samples were found positive for NSP antibodies indicating infection. In LPB ELISA, 42 of 48 (87.5%) pigs were found to have protective log10 antibody titre of ≥1.8 against FMD virus serotypes O, A and Asia 1. All the clinical materials were found positive for serotype O in antigen detection ELISA as well as in multiplex reverse transcription-polymerase chain reaction (mRT-PCR). In VP1 region-based phylogenetic analysis, the serotype O isolates causing the outbreak were found to conglomerate within Ind2001 lineage. Pigs infected with FMD may pose rigorous threat to other susceptible domestic livestock as they exhale enormous quantity of virus. As a consequence, they should be included under prophylactic vaccination and surveillance programmes ongoing in the country. [ABSTRACT FROM AUTHOR]

Summary: Foot‐and‐mouth disease (FMD) is an important transboundary disease with substantial economic impacts. Although between‐herd transmission of the disease has been well studied, studies focusing on within‐herd transmission using farm‐level outbreak data are rare. The aim of this study was to estimate parameters associated with within‐herd transmission, host physiological factors and FMD virus (FMDV) persistence using data collected from an outbreak that occurred at a large, organized dairy farm in India. Of 1,836 regularly vaccinated, adult dairy cattle, 222 had clinical signs of FMD over a 39‐day period. Assuming homogenous mixing, a frequency‐dependent compartmental model of disease transmission was built. The transmission coefficient and basic reproductive number were estimated to be between 16.2–18.4 and 67–88, respectively. Non‐pregnant animals were more likely to manifest clinical signs of FMD as compared to pregnant cattle. Based on oropharyngeal fluid (probang) sampling and FMDV‐specific RT‐PCR, four of 36 longitudinally sampled animals (14%) were persistently infected carriers 10.5 months post‐outbreak. There was no statistical difference between subclinical and clinically infected animals in the duration of the carrier state. However, prevalence of NSP‐ELISA antibodies differed significantly between subclinical and clinically infected animals 12 months after the outbreak with 83% seroprevalence amongst clinically infected cattle compared to 69% of subclinical animals. This study further elucidates within‐herd FMD transmission dynamics during the acute‐phase and characterizes duration of FMDV persistence and seroprevalence of FMD under natural conditions in an endemic setting. [ABSTRACT FROM AUTHOR]

Summary: The goal of this study was to characterize the properties and duration of the foot‐and‐mouth disease (FMD) carrier state and associated serological responses subsequent to vaccination and naturally occurring infection at two farms in northern India. Despite previous vaccination of cattle in these herds, clinical signs of FMD occurred in October 2013 within a subset of animals at the farms containing juvenile‐yearling heifers and steers (Farm A) and adult dairy cattle (Farm B). Subsequent to the outbreak, FMD virus (FMDV) asymptomatic carriers were identified in both herds by seroreactivity to FMDV non‐structural proteins and detection of FMDV genomic RNA in oropharyngeal fluid. Carriers’ seroreactivity and FMDV genome detection status were subsequently monitored monthly for 23 months. The mean extinction time of the carrier state was 13.1 ± 0.2 months, with extinction having occurred significantly faster amongst adult dairy cattle at Farm B compared to younger animals at Farm A. The rate of decrease in the proportion of carrier animals was calculated to be 0.07 per month. Seroprevalence against FMDV non‐structural proteins decreased over the course of the study period, but was found to increase transiently following repeated vaccinations. These data provide novel insights into viral and host factors associated with the FMDV carrier state under natural conditions. The findings reported herein may be relevant to field veterinarians and governmental regulatory entities engaged in FMD response and control measures. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) is a highly contagious and globally significant viral disease principally of cloven-hoofed livestock species. The present study describes the results of molecular epidemiologic investigation of FMD in pigs across various states of India between 2008 and 2014. During this period, a total of 37 clinical epithelial samples (vesicle/foot/snout epithelium) of FMD-suspected pigs were tested in a serotype differentiating antigen detection ELISA and samples found negative were further subjected to multiplex reverse transcription-polymerase chain reaction (mRT-PCR). A total of 29 (78.37%) samples were found positive for serotype O in antigen detection ELISA and 8 ELISA-negative samples were subsequently found positive for serotype O in mRT-PCR. The VP1 region-based phylogenetic analysis demonstrated the involvement of O/ME-SA/Ind2001 lineage in the outbreaks. The pig isolates clustered with the contemporary virus isolates collected from bovine indicating a close genetic relationship and therefore signifying inter-species transmission during the outbreaks. [ABSTRACT FROM AUTHOR]

The yak and mithun husbandry in India is confronted with several challenges including the prevalence of foot-and-mouth disease (FMD). The present study was initiated to investigate FMD outbreaks in semi-domesticated mithun and yak population in various parts of India. A total of 64 clinical samples (vesicle/tongue/foot epithelium/fluid) from mithun and 6 from yak were collected from suspected FMD outbreaks during 2008-2013. Supernatants of the homogenized clinical samples were tested in a serotype discriminating antigen detection ELISA and ELISA-negative samples were further subjected to multiplex reverse transcription-polymerase chain reaction (mRT-PCR). A total of 45 mithun samples and only 1 yak sample were found positive for serotype O in antigen detection ELISA. A total of 12 ELISA-negative samples from mithun and 4 from yak were later on found positive for serotype O in mRT-PCR. The phylogenetic analysis based on VP1 genome indicated the involvement of both O/ME-SA/PanAsia and O/ME-SA/Ind2001 lineages of serotype O in those outbreaks. These viruses were genetically similar to those contemporary virus isolates responsible for FMD in domestic livestock indicating a situation of virus sharing among different species of domestic and semi-domestic animals. Thus, mithuns and yaks should be synchronously considered and targeted along with cattle for the effective control of the disease in the country. [ABSTRACT FROM AUTHOR]

A systematic vaccination programme is ongoing in India to control the three prevailing serotypes (A, O, Asia1) of foot-and-mouth disease ( FMD) virus. Under the programme, more than 120 million bovine (term bovine applicable to both cattle and buffalo in this study) population of 221 of the 666 districts in the country are being bi-annually vaccinated with trivalent vaccine since 2010. Although clinical disease has reduced in these districts because of the systematic vaccinations, an abrupt increase in the number of FMD cases was recorded in 2013. Hence, a longitudinal field study was conducted in the year 2014 to estimate the serological herd immunity level in bovines, the impact of systematic vaccinations and field efficacy of the vaccines used. Serum samples ( n = 115 963) collected from 295 districts of the 18 states of the country were analysed to estimate antibody titres against structural proteins of the three serotypes. The efficacy of the vaccine was demonstrated in the control group (group-D) where animals of the group were identified by ear tags for the purpose of repeated sampling after vaccination. Progressive building of the herd immunity in the field after systematic vaccination was demonstrated. The mean antibody titre against the serotypes O, A and Asia1 was estimated as log10 1.93 (95% CI 1.92-1.93), 2.02 (2.02-2.02) and 2.02 (2.02-2.02), respectively, in the states covered under the control programme. However, in other states herd immunity was significantly low [mean titre log10 1.68 (95% CI 1.67-1.69), 1.77 (1.76-1.78) and 1.85 (1.84-1.86) against the three serotypes]. Inverse relationship between the herd immunity and FMD incidences was observed the states following different vaccination practices. The study helped in demarcation of FMD risk zones in the country with low herd immunity. Estimation of herd immunity kinetics in the field helped in refining the vaccination schedule under the control programme. [ABSTRACT FROM AUTHOR]

This study was conducted to assess the impact of Foot-and-mouth Disease ( FMD) outbreak in cattle and buffaloes on farming community in Kolar district, Karnataka state, India. Primary data were collected using pre-tested schedule from 178 sample farms using multistage random cluster sample technique. The results revealed that 78% of surveyed villages were affected with FMD. The FMD incidence risk was high across the herd sizes, whereas the mortality risk was high in small herds. In indigenous cattle, the highest loss due to FMD was distress sale (208 USD) followed by other losses, whereas, in Crossbred cattle, the highest loss was mortality loss (515 USD) followed by distress sale (490 USD), milk yield loss (327 USD), treatment cost (38 USD) and extra labour engagement expenses for nursing of FMD-affected bovines (30 USD). In local and upgraded buffaloes, the mean total loss per affected animal was 440 USD and 513 USD, respectively. A very high variability in the loss per animal was observed across the type of losses in the Crossbred cattle, and it may be due to differences in age of the FMD-infected animal, value of the animal, milking stage, lactation levels, herd sizes and labour engagement levels, etc. In local and upgraded buffaloes, the mean total loss per animal was 639 USD and 1008 USD, respectively. The sensitivity analysis for 5% change in price revealed that the mean total loss per animal was positively correlated with price. Further, the social impact elicitation revealed that majority of the livestock owners perceived FMD had caused permanent asset loss, which in turn increased psychological stress of the family. The estimated losses and social impact due to FMD signify the importance of the intervention to control the disease and thus socio-economic gain to the farmer and society at large. [ABSTRACT FROM AUTHOR]

Small ruminants (sheep and goats) are susceptible to foot-and-mouth disease (FMD), while studies with due emphasis on their role in the disease epidemiology have been meagre. The present study summarizes the results of clinico-molecular investigation of FMD in sheep and goats across several states of India during 2008–2014, where a total of 51 clinical epithelial tissue samples (vesicle/tongue/gum/foot epithelium) from sheep and 78 from goats were found positive for FMD virus (FMDV) serotype O in serotyping ELISA and multiplex reverse transcription-PCR. The VP1 region-based phylogenetic analysis demonstrated the involvement of O/ME-SA/Ind2001 lineage of serotype O virus in the outbreaks. The field viruses recovered from both small and large ruminant population during the same time period showed a close genetic relationship suggesting frequent inter-species transmission of virus. Since the disease often remains clinically camouflaged in small ruminants, the animals silently infected with FMD may pose potential threats to the in-contact livestock. Regular vaccination combined with surveillance and monitoring of protective antibody status in these species is therefore crucial to the effective control of FMD in the country. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease viruses are often restricted to specific geographical regions and spread to new areas may lead to significant epidemics. Phylogenetic analysis of sequences of the VP1 genome region of recent outbreak viruses from Libya and Saudi Arabia has revealed a lineage, O-Ind-2001, normally found in the Indian subcontinent. This paper describes the characterization of field viruses collected from these cases and provides information about a new real-time RT- PCR assay that can be used to detect viruses from this lineage and discriminate them from other endemic FMD viruses that are co-circulating in North Africa and western Eurasia. [ABSTRACT FROM AUTHOR]

Summary: Porcine sapelovirus (PSV) A belongs to the genus Sapelovirus, family Picornaviridae. PSV infections in pigs have been reported from European countries, United States, Japan, China, Korea and Brazil. The virus has been isolated/detected from faeces of healthy pigs as well as those affected with diarrhoea, respiratory signs, encephalitis, skin lesions and fertility disorders. This study was planned to investigate whether PSV is prevalent among pigs in India and to characterize PSV encountered in the study population. The study revealed that five of 70 (7.14%) faecal samples were found positive for PSV using RT‐PCR. Three viruses were successfully isolated from faecal samples using IB‐RS‐2 cell line. Complete genome sequencing and analysis of one Indian PSV isolate revealed highest homology (88%) with V13 strain from England. Phylogenetic analysis of the complete polyprotein nucleotide sequences of 14 strains of PSV classified the viruses into four distinct clades. This first report from India adds to our knowledge on genetic diversity of PSV detected so far among pigs in different countries. A large‐scale surveillance of the virus is required to understand its genomic diversity and economic impact. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) virus serotype O is the most common cause of FMD outbreaks in India and three of the six lineages that have been described are most frequently detected, namely Ind2001, PanAsia and PanAsia 2. We report the full capsid sequence of 21 serotype O viruses isolated from India between 2002 and 2012. All these viruses belong to the Middle East–South Asia (ME–SA) topotype. The serological cross-reactivity of a bovine post-vaccination serum pool raised against the current Indian vaccine strain, O/IND/R2/75,was tested by virus neutralisation test with the 23 Indian field isolates, revealing a good match between the vaccine and the field isolates. The cross reactivity of the O/IND/R2/75 vaccine with 19 field isolates from other countries (mainly from Asia and Africa) revealed a good match to 79% of the viruses indicating that the vaccine strain is broadly cross-reactive and could be used to control FMD in other countries. Comparison of the capsid sequences of the serologically non-matching isolates with the vaccine strain sequence identified substitutions in neutralising antigenic sites 1 and 2, which could explain the observed serological differences. [ABSTRACT FROM AUTHOR]

A total of 41 clinical samples (vesicle/tongue/foot/nasal epithelium) from Indian gaur, deer, spotted deer, nilgai, chowsinga, bison, black buck, elephant, sambar deer were collected in 50% phosphate buffered saline/glycerol medium (pH-7.5) during suspected FMD outbreaks. Supernatants of homogenized clinical samples were used in a serotype differentiating antigen detection ELISA and samples found negative were further subjected to multiplex PCR (mPCR). A total of 3/11 (27.2%) samples from Indian gaur, 2/7 (28.5%) chital deer, 5/5 (100%) nilgai, 2/2 (100%) black buck were found positive for serotype O in antigen detection ELISA. A total of 3 ELISA-negative samples from spotted deer, 2 from bison and 2 from sambar deer were found positive for serotype O in mPCR. The VP1 region-based phylogenetic analysis indicated the involvement of both O/ME-SA/Ind2001 and PanAsia lineage of serotype O in the outbreaks. The wildlife species infected with FMD may pose further threat to the surrounding domestic livestock. [ABSTRACT FROM AUTHOR]

Foot and mouth disease (FMD) is an economically important disease and a whole-virus inactivated trivalent virus vaccine is the mainstay for controlling the disease in India. The protective humoral immune response to FMD vaccination is a complex, but, tightly regulated process mediated by the interplay of interleukins (IL). Based on the specific role of IL6 and 21 in adaptive immune response, we hypothesized that inactivated trivalent FMD vaccine would stimulate IL6 and 21 expression in the circulating lymphocytes. The expressions of IL6 and 21 were assayed on 0, 28, 60, 90, and 120 d post-vaccination (DPV) by quantitative PCR (qPCR) with simultaneous assessment of FMDV antibody titer by liquid phase blocking ELISA. The results revealed that the peak expression of IL6 and 21 was on DPV 28 which correlated well with the FMDV antibody titer and plummeted to the prevaccination titer level by 60 DPV. As IL21 is the final effector of antibody production as compared to IL6, we investigated the expression of IL21 in calves that had protective titer (>1.8) with the unprotected group (<1.8). Expression of IL21 on 28 DPV was numerically higher in the protected than that of the unprotected group of calves. [ABSTRACT FROM AUTHOR]

Polymorphism of bovine leukocyte antigen (BoLA) DRB3 gene is being intensively investigated for potential association with economically important diseases of cattle. Accordingly, we investigated the association of DRB3 Exon 2 polymorphism as evidenced by the variation in the binding pockets with variability in immune response to inactivated trivalent (O, A and Asia1) foot and mouth disease virus (FMDV) vaccine in a closed population of crossbred cattle. Antibody titer of ⩾1.8 was set as the cut off value to distinguish the protected (P1.8) and unprotected (⩾1.8) animals. Eleven different alleles of over 3% frequency were detected in the population. We found that DRB3 alleles 8 0201, 8 0801 and 8 1501 always ranked high for protective immune response whereas alleles 8 0701, 8 1103 and 8 1101 consistently ranked low for unprotected immune response for all the three serotypes. Rank correlation of DRB3 alleles among the three serotypes was positive, high in magnitude and statistically significant (P < 0.05). Logistic regression analysis revealed that odds of protection from the vaccine were highest for all the three serotypes if allele 8 1501 was present and strengthened the results of allele ranking. Predicted amino acid substitution in the peptide binding pockets revealed that all the important sites had high Wu-Kabat index. Similarly, specific residues in pockets were crucial for immune response to FMD vaccine. There were specific substitutions in un-protected alleles such as absence of acidic amino acids substituted by basic amino acid at β71, presence of non-polar cysteine or basic histidine at b30 and presence of polar tyrosine at β37. From the observations, we hypothesize that the substitutions lead to unique conformational changes in the protein products of the studied alleles that would associate with the protective or unprotective antibody response to FMDV vaccine. The knowledge has potential implications in future selection programs if integrated with the complete BoLA haplotype details and production traits of the herd. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the endometrial expression of pro-inflammatory cytokines (IL1β, IL6, IL8 and TNFα) along with TLR4 and CD14 in normal and endometritic buffaloes. The genitalia were collected in the abattoir and divided into three groups as normal (gr. I = 12), clinical endometritis (CE, gr. II = 12) based on positive color reaction to white side test of uterine discharge and sub-clinical endometritis (SCE, gr. III = 12) based on endometrial cytology (presence of ⩾5% PMNs) and histopathology. The equal numbers of genitalia were grouped into follicular and luteal stage in each group. Endometrial tissue scrapings were used for total RNA extraction and cDNA was transcribed and amplified by Real time PCR. The results showed several fold higher expression of all cytokine transcripts in CE (gr. II), whereas significant up-regulation of CD14 (1 to 2-fold), IL6 (15 to 36-fold), IL8 (8 to 14-fold) and TNFa (10 to 11-fold) mRNA was observed in SCE. This indicates that the evaluation of expression patterns of certain cytokines gene holds promise to diagnose the severity and degree of uterine inflammation. [ABSTRACT FROM AUTHOR]

India is endemic for foot-and-mouth disease (FMD), and goats constitute the second largest susceptible population of domestic livestock. FMD surveillance and control strategies in the country largely ignore small ruminants, known to be critical in the epidemiology of the disease. Here, serological investigations were carried out to generate estimates of antibody prevalence in goats of Orissa state to both non-structural (NSP-Ab) and structural proteins (SP-Ab) of FMD. The apparent overall NSP-Ab and SP-Ab seroprevalences were 38% and 20.7%, respectively, which signifies a very high level of FMD virus circulation in the goat population despite the lack of clinical signs in this species. The apparent prevalence of NSP-Ab and SP-Ab was positively correlated in the sampling areas. Interestingly, the values found for NSP-Ab prevalence were almost consistently higher than those found for SP-Ab prevalence. This could have been attributable to either issues related to sensitivity and specificity of the test systems employed or differences in the post-infection kinetics of NSP- and SP-Ab. The pattern that emerged from SP-Ab analysis indicated goats being infected with all three prevalent serotypes (O, A and Asia 1) and reinforces the concept that non-vaccinated goats can be exploited as tracer animals for detecting serotypes involved in outbreaks. The results underscore the requirement to bring caprine species under comprehensive surveillance and vaccination campaigns to check silent amplification, excretion and transmission of the virus. [ABSTRACT FROM AUTHOR]

Background & objectives: With the emergence of a new reassortant influenza A H1N1 virus that caused the 2009 pandemic it was felt necessary that pigs should be closely monitored for early detection of any influenza virus infection. Therefore, we investigated disease outbreaks with clinical history suggestive for swine influenza reported to our laboratory by owners of affected pig farms in Uttar Pradesh. Methods: Detection of swine influenza A virus (SIV) was attempted by isolation in embryonated chicken eggs. Presence of virus was detected by haemagglutination (HA) test and RT-PCR for amplification of different gene segments, cloning and sequencing. BLAST analysis of sequence data, phylogenetic analysis and mutation analysis based on HA, NA and matrix genes was done. Results: SIV could be isolated from one farm and all eight gene segments amplified by RT-PCR. BLAST analysis of partial nucleotide sequences and phylogenetic analysis using nucleotide sequence of HA (601 nt), NA (671 nt) and M (1031 nt) genes indicated close genetic relationship of the Indian swine isolate (A/ Sw/UP-India-IVRI01/2009) with human pandemic 2009 (H1N1). The HA gene showed close relationship with the viruses of "North American Swine" lineage, whereas the NA and M genes clustered with the viruses of "Eurasian Swine" lineage, indicating a novel HA-NA reassortant. The remaining of 5 genes (NP, PA, PB1, PB2 and NS) belonged to "North American Swine" lineage. Interpretation & conclusions: This is perhaps the first report describing swine influenza among Indian pigs caused by an influenza A H1N1 virus sharing close homology with the human pandemic (H1N1) 2009 virus. Further reassortment with circulating influenza viruses must be closely monitored. [ABSTRACT FROM AUTHOR]

We investigated the molecular epidemiology of foot-and-mouth disease virus (FMDV) serotype Asia 1, which caused outbreaks of disease in Asia during 2003-2007. Since 2004, the region affected by outbreaks of this serotype has increased from disease-endemic countries in southern Asia (Afghanistan, India, Iran, Nepal, Pakistan) northward to encompass Kyrgyzstan, Tajikistan, Uzbekistan, several regions of the People's Republic of China, Mongolia, Eastern Russia, and North Korea. Phylogenetic analysis of complete virus capsid protein 1 (VP1) gene sequences demonstrated that the FMDV isolates responsible for these outbreaks belonged to 6 groups within the Asia 1 serotype. Some contemporary strains were genetically closely related to isolates collected historically from the region as far back as 25 years ago. Our analyses also indicated that some viruses have spread large distances between countries in Asia within a short time. [ABSTRACT FROM AUTHOR]

Abstract: A total of 1246 faecal and tissue samples collected/received from 119 farms located in various states of India were processed for isolation of avian influenza viruses (AIV) during 2003–2004 as part of a program to monitor AIV infection in Indian poultry population. Avian influenza virus was isolated for the first time in India from poultry farms with history of drop in egg production, respiratory illness and increased mortality in Haryana state. A total of 29 H9N2 AIV isolates were obtained from the states of Punjab, Haryana, Uttar Pradesh, Gujarat, and Orissa and Union Territory Delhi. Subtyping was done by HI, RT-PCR and neuraminidase inhibition assay. Pathotyping of six representative isolates by intravenous pathogenicity index (0.0/3.0) in 6–8 weeks old chicken, trypsin dependency in cell culture and HA cleavage site analysis (335RSSR*GLF341) confirmed that these isolates are low pathogenic. Nucleotide sequence analysis of the HA gene showed that the Indian isolates are very closely related (95.0–99.6%) and shared a homology of 92–96% with H9N2 isolates from Germany and Asian regions other than that of mainland China. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) which indicates a preference to binding of α (2–6) sialic acid receptors. Two of the six isolates had 7 glycosylation sites in the HA1 cleaved protein and the remaining four had 5 sites. Phylogenetic analysis showed that they share a common ancestor Qa/HK/G1/97 isolate which had contributed internal genes of H5N1 virus circulating in Vietnam. Further characterization of Indian H9N2 isolates is required to understand their nature and evolution. [Copyright &y& Elsevier]

Abstract: The aim of this study was to develop a competitive inhibition ELISA (CI-ELISA) for detection of antibodies to bovine viral diarrhea virus (BVDV) using the helicase domain of NS3 (non-structural) protein and monoclonal antibody (MAb) against it and to estimate its sensitivity and specificity using two commercial ELISA kits as independent references. The 45-kDa helicase domain of NS3 protein of BVDV was expressed in Escherichia coli and 18MAbs were developed against it. MAb-11G8 was selected for use in CI-ELISA on the basis of maximum inhibition (90%) obtained with BVDV type 1 infected calf serum. Based on the distribution of percent inhibition of known negative sera (n =166), a cut-off value was set at 40% inhibition. In testing 914 field serum samples of cattle (810) and buffaloes (104), the CI-ELISA showed a relative specificity of 95.75% and 97.38% and sensitivity of 96% and 94.43% with Ingenesa kit and Institut Pourquier kit, respectively. This study proved that the use of helicase domain of NS3 (45-kDa) is equally good as the whole NS3 protein (80-kDa) used in commercial kits for detection of BVDV antibodies in cattle and buffaloes. [Copyright &y& Elsevier]

H9N2 avian influenza viruses are endemic in domestic poultry in Asia and are grouped into three major sublineages represented by their prototype strains A/Duck/Hong Kong/Y280/97 (Y280-like), A/Quail/Hong Kong/G1/97 (G1-like) and A/Chicken/Korea/38349-p96323/96 (Korean-like). To understand the genetic relationship of Indian viruses, we determined the partial nucleotide sequence of five H9N2 avian influenza viruses isolated from chicken in India during 2003–2004 and compared them with H9N2 sequences available in GenBank. Deduced amino acid sequence analysis revealed that four isolates shared an R–S–S–R/G motif at the cleavage site of HA, representing low pathogenicity in chickens, while one virus harbors an R–S–N–R/G motif at the same position. All the viruses maintained the human-like motif 226Lysine (H3 numbering) at the HA receptor binding site. Phylogenetic analysis showed that 50% of the genes (HA, NA, NP and M) were similar to G1-like viruses, whereas the remaining genes of the Indian isolates formed a separate, not yet defined, sublineage in the Eurasian lineage. Our finding provides evidence of a novel reassortant H9N2 genotype of G1-like viruses circulating in India. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to develop a more specific and sensitive competitive inhibition ELISA (CI-ELISA) than the currently used indirect ELISA for detection of antibodies to bovine immunodeficiency virus (BIV) in cattle and buffaloes. Murine monoclonal antibodies (MAbs) were generated against a recombinant capsid (CA) protein of bovine immunodeficiency virus. Of the 13 anti-CA MAbs developed, MAb-9G10 was selected for CI-ELISA based on the maximum inhibition (98%) obtained with reference BIV antibody positive serum. Based on the distribution of percent inhibition of known negative sera (n =50), a cut-off value was set at 40% inhibition. The MAb-based CI-ELISA showed much higher agreement (concordance: 95.4%) than the indirect ELISA (concordance: 77.8%) with Western blot. Out of 672 sera of cattle and buffaloes tested by CI-ELISA from four states of India, 22% (113/516) of cattle and 19% (30/156) of buffalo were sero-positive for BIV with an overall seroprevalence of 21% (143/672) in India. [Copyright &y& Elsevier]

Abstract: One-step SYBR Green I-based real-time RT-PCR assay for rapid detection as well as quantitation of Japanese encephalitis virus (JEV) in acute-phase patient CSF samples by targeting the NS3 gene was developed. The assay developed in this study was found to be more sensitive as compared to conventional RT-PCR. The specificity of the reported assay system was established through melting curve analysis as well as by cross-reactivity studies with related members of Flavivirus. The applicability of Real-time PCR assay for clinical diagnosis was validated with 32 suspected acute-phase CSF samples of Gorakhpur epidemic, India, 2005. The improved sensitivity of real-time RT-PCR was reflected by picking up 10 additional samples with low copy number of template in comparison to conventional RT-PCR. The quantitation of the viral load in acute-phase CSF samples was done using a standard curve obtained by plotting cycle threshold (C t) values versus copy numbers of the RNA template. This is the first report on the application of real-time RT-PCR for detection as well as quantitation of JEV from patient CSF samples. These findings demonstrate the potential clinical application of the reported assay as a sensitive diagnostic test for rapid and real-time detection and quantitation of JEV in acute-phase clinical samples. [Copyright &y& Elsevier]

Abstract: The development of a one-step SYBR Green I-based real-time RT-PCR assay is reported for detection and quantification of Chikungunya virus (CHIKV) in acute-phase patient serum samples by targeting the E1 structural gene. A linear relationship was obtained between the virus concentration and cycle threshold (C t) value over a range of 107–0.1PFU/ml. The reported assay was found to be 10-fold more sensitive compared to conventional RT-PCR with a detection limit of 0.1PFU/ml. The feasibility of this reported assay system for clinical diagnosis was validated with 51 suspected acute-phase serum samples of the recent CHIKV epidemic in southern India, 2006. The comparative evaluation with acute-phase patient serum samples revealed the higher sensitivity of real-time RT-PCR assay by picking up six additional samples with low copy number of template. None of the healthy serum samples analyzed in this study showed amplification. The quantification of the viral load in the acute-phase serum samples was also determined employing the standard curve, which varies from 0.1 to 107 PFU/ml. These findings demonstrated that the reported assay has the potential usefulness for clinical diagnosis due to simultaneous detection and quantification of Chikungunya virus in acute-phase patient serum samples. [Copyright &y& Elsevier]

Abstract: India is one of the countries with the highest prevalence of human rabies throughout the world. Dogs are primarily responsible for rabies transmission. Among them, stray dogs play a major role in that country. Parenteral vaccination programmes are insufficient to eliminate rabies partly due to difficulties in establishing satisfactory immunisation coverage in the dog population in view of the high proportion of stray dogs. Oral vaccination may be a useful adjunct to parenteral vaccination by increasing dog vaccination coverage. Safety, immunogenicity and efficacy of Rabidog® SAG2 bait were evaluated in Indian stray dogs in captivity. Safety of SAG2 was demonstrated by the absence of adverse clinical sign, salivary excretion and absence of replication of the vaccine strain in brain and salivary glands of 21 vaccinated dogs, even when immunodepressed. Efficacy was shown 109 days post-vaccination after challenge with a highly virulent street rabies virus which killed all five controls whereas all nine vaccinated dogs survived, despite the fact that only five out of nine had seroconverted before challenge. [Copyright &y& Elsevier]

Complete 1D gene sequences of 13 Indian foot-and-mouth disease virus (FMDV) type C field isolates and a vaccine strain (C-Bombay/64) were determined. All the field isolates showed a greater genetic homogeneity (95–100%) among themselves and were 19.7–21.2% divergent from the vaccine strain. In the phylogenetic analysis, the Indian field isolates formed a separate lineage (lineage VII) different from the previously identified six lineages (lineage I–VI) in type C FMDV [J. Virol. 66 (1992) 3557]. The vaccine strain was grouped with European lineage (lineage II). Comparison of the deduced amino acid sequences of antigenic sites A and C of field isolates showed no significant variation from the vaccine strain. One-way serological relationship determined in ELISA showed antigenic closeness of the field isolates with C-Bombay/64. [Copyright &y& Elsevier]

Abstract: In India, systematic genotype replacement has been observed for serotype A foot-and-mouth disease virus. After a decade of co-circulation of genotypes VI and VII, genotype VII emerged as the single dominant genotype since 2001. To derive possible explanations for such epochal evolution dynamics, in vitro intergenotype growth competition experiments involving both co- and superinfection regimes were conducted. Coinfection of BHK-21 cells demonstrated abrupt loss in the genotype VI viral load with commensurate increase in the load of genotype VII as measured by the genotype differentiating ELISA, RT-PCR and real-time RT-PCR. The superinfection dynamics was shaped by temporal spacing of infection, where the invading genotype VII took more number of passages than coinfection to eventually overtake the resident genotype VI. It was speculated that such superior replicative fitness of genotype VII could have been a possible factor for the ultimate dominance of genotype VII in nature. [Copyright &y& Elsevier]