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Foot-and-mouth disease Virus (FMDV) serotype Asia1 is prevalent in the Indian subcontinent, with only G-III and G-VIII reported in India until 2020. However, in 2019, a novel genetic group within serotype Asia1, designated as G-IX, emerged in Bangladesh, followed by its detection in India in 2020. This report presents analyses of the complete coding region sequences of the G-IX lineage isolates. The length of the open reading frame (ORF) of the two G-IX isolates was 6990 nucleotides without any deletion or insertion. The G-IX isolates showed the highest sequence similarity with an isolate of G-III at the ORF, L, P2, and P3 regions, and with an isolate of G-VIII at the P1 region. Phylogenetic analysis based on the capsid region (P1) supports the hypothesis that G-VIII and G-IX originated from a common ancestor, as speculated earlier. Further, VP1 region-based phylogenetic analyses revealed the re-emergence of G-VIII after a gap of 3 years. One isolate of G-VIII collected during 2023 revealed a codon insertion in the G-H loop of VP1. The vaccine matching studies support the suitability of the currently used Indian vaccine strain IND63/1972 to contain outbreaks due to viruses belonging to G-IX.

In the present study, tissue samples (tongue, esophagus and heart) were investigated from dromedary camels of India for identification and characterization of Sarcocystis spp. using histopathology, PCR and gene sequencing. Genomic DNA extracted from these tissue samples was used for PCR amplification of the cytochrome c oxidase subunit I gene (cox1) of Sarcocystis spp. and the partial sequence of small subunit ribosomal RNA (18S rRNA) gene of the S. cameli. The PCR products were purified, sequenced and analyzed using bioinformatics tools. Based on phylogenetic analysis of the cox1 gene, the sequences of the present study clustered with those of S. cameli, hosted by dromedary camels of Iraq and a close association was observed with S. masoni hosted by dogs and alpacas of China. Until now, there are no 18S rRNA sequences of S. cameli available in GenBank and this is the first study recording 18S rRNA sequences of S. cameli which were grouped with S. masoni from alpaca of China and guanaco and llama of Argentina in phylogenetic analysis. These findings could be useful for further studies on the characterization through molecular epidemiology, genetic diversity and host specificity of S. cameli., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Serotype identification occupies the central part of foot and mouth disease (FMD) diagnosis workflow and vaccination decision tree. In this study, a reverse transcription-multiplex PCR (RT-mPCR) strategy wherein three assays with unique combinations of serotype specific primers targeting the VP1 region was developed to differentiate FMD virus serotypes O, A and Asia 1 based on differential size of the PCR amplicons on agarose gel. Their diagnostic performance relative to the mPCR assay in use in India was evaluated on 169 clinical samples and 210 cell culture grown virus isolates. The relative diagnostic sensitivity was found to be 99.69%, 98.78% and 99.08% for primer combinations 1, 2 and 3, respectively. These assays proved their worth by detecting serotype in three FMD suspected specimens that went undiagnosed in the existing mPCR and also by identifying multiple serotypes in the same sample. Their detection limits varied from log 10 2 to log 10 4 viral RNA dilution and from 100 to 0.1 TCID 50 virus depending on the serotype. The validated novel mPCR assays show promise to be included in the routine diagnostic tool-box to augment the efficiency of diagnosis of FMD virus serotypes that display extreme genetic diversity and a tendency of transboundary dispersal., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Foot and mouth disease (FMD) has engendered large scale socioeconomic crises on numerous occasions owing to its extreme contagiousness, transboundary nature, complicated epidemiology, negative impact on productivity, trade embargo, and need for intensive surveillance and expensive control measures. Emerging FMD virus variants have been predicted to have originated and spread from endemic Pool 2, native to South Asia, to other parts of the globe. In this study, 26 Indian serotype A isolates sampled between the year 2015 and 2022 were sequenced for the VP1 region. BLAST and maximum likelihood phylogeny suggest emergence of a novel genetic group within genotype 18, named here as 'A/ASIA/G-18/2019' lineage, that is restricted so far only to India and its eastern neighbour, Bangladesh. The lineage subsequent to its first appearance in 2019 seems to have displaced all other prevalent strains, in support of the phenomenon of 'genotype/lineage turnover'. It has diversified into two distinct sub-clusters, reflecting a phase of active evolution. The rate of evolution of the VP1 region for the Indian serotype A dataset was estimated to be 6.747 × 10 -3 substitutions/site/year. India is implementing a vaccination centric FMD control programme. The novel lineage showed good antigenic match with the proposed vaccine candidate A IND 27/2011 when tested in virus neutralization test, while the existing vaccine strain A IND 40/2000 showed homology with only 31% of the isolates. Therefore, in order to combat this challenge of antigenic divergence, A IND 27/2011 could be the preferred strain in the Indian vaccine formulations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financialinterestsor personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

In India, widespread foot-and-mouth disease (FMD) outbreaks occurred in 2021. The objective of this study was to identify genetic lineages and evaluate the antigenic relationships of FMD virus (FMDV) isolates gathered from outbreaks reported between 2019 and 2022. Our study shows that the lineages O/ME-SA/Ind2001e and the O/ME-SA/Cluster-2018 were both responsible for the FMD outbreaks on an epidemic scale during 2021. This observation is in contrast to earlier findings that suggested epidemic-scale FMD outbreaks in India are often connected to a single genetic lineage. Additionally, we report here the identification of the O/ME-SA/PanAsia-2/ANT10 sub-lineage in India for the first time, which was connected to two intermittent outbreaks in Jammu and Kashmir. The current study demonstrates that the O/ME-SA/ind2001e lineage has a strong presence outside of the Indian subcontinent. Furthermore, the O/ME-SA/Cluster-2018 was observed to have a wider geographic distribution than previously, and like the O/ME-SA/Ind2001d and O/ME-SA/Ind2001e lineages in the past, it may eventually spread outside of its geographic niche. For O/ME-SA/Ind2001e and O/ME-SA/Cluster-2018, the predicted substitution rate for the VP1 region was 6.737 × 10 -3 and 8.257 × 10 -3 nt substitutions per site per year, respectively. The time of the most recent common ancestor of the O/ME-SA/Ind2001e and O/ME-SA/Cluster-2018 strains suggests that the viruses possibly emerged during 2003-2011 and 2009-2017, respectively. Recent sightings of the O/ME-SA/PanAsia2/ANT10 virus in India and the O/ME-SA/Ind2001e virus in Pakistan point to possible cross-border transit of the viruses. The results of a two-dimensional viral neutralization test revealed that all of the field isolates were antigenically matched to the currently used Indian vaccine strain O INDR2/1975. These results suggest that the serotype O vaccine strain can protect against outbreaks brought on by all three circulating lineages.

Foot-and-mouth disease (FMD) is a major disease of livestock in India and causes huge economic losses. The formal FMD control program started in 2003-04 in selected districts and was gradually expanded. The present study provides a descriptive review of the FMD outbreaks, prevalent serotypes, and genetic and antigenic features of the FMD virus (FMDV) that circulated in the country between 2011 and 2020. FMD outbreaks were regularly reported in cloven-hoofed domestic livestock and wildlife, with three serotypes including O, A, and Asia1. During the study period, a total of 2226 FMD outbreaks were documented and serotypes confirmed. FMDV serotype O dominated the outbreak scenario, accounting for about 92% of all outbreaks, followed by Asia1 (5% of all outbreaks) and A (3% of all outbreaks). Two major epidemics of FMD on an unprecedented scale during the years 2013 and 2018 by serotype O were recorded. The spatial distribution of FMD was characterized by a larger number of outbreaks in the southern region of the country. In an annual-scale analysis, 2020 was the year with the lowest outbreaks, and 2013 was the year with the highest. The month-scale analysis showed that outbreaks were reported throughout the year, with the highest numbers between October and March. The emergence of three major lineages (O/ME-SA/Ind2001d, O/ME-SA/Ind2001e, and O/ME-SA/Ind2018) of serotype O was observed during the period. In the cases of serotype A and Asia1, the appearance of at least one novel lineage/genetic group, including A/G-18/non-deletion/2019 and Asia1/Group-IX, was documented. While serotype A showed the advent of antigenic variants, serotypes O and Asia1 did not show any antigenic diversity. It was noticed during the course of an outbreak that animal movement contributes significantly to disease transmission. Except for 2018, when numerous FMD outbreaks were recorded, the number of annual outbreaks reported after 2016 has been lower than in the first half of the decade, probably due to mass vaccination and COVID-19 pandemic-linked movement restrictions. Even during outbreaks, disease symptoms in ruminant populations, including cattle, were found to be less severe. Regular six-monthly immunization certainly has a positive impact on the reduction of disease burden and should be followed without fail and delay, along with intensive disease surveillance., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

The focus on the bioactive effects of plants concerns mainly ruminal microflora for the modulation of rumen fermentation with very little emphasis placed on their consequences on health parameters, including antioxidative and immune stimulating effects. The aim of the present study was to examine the effects of supplementing phytogenic feed additives composed of a mixture of eucalyptus ( Eucalyptus citriodora) and poplar ( Populus deltoides ) leaf-meal (EPLM) on immune response, antioxidant status, metabolic profile, enteric methane production, growth performance, and nutrient utilization in buffalo ( Bubalus bubalis ) calves. In vitro studies with graded doses of EPLM extract revealed a reduction in total gas and methane production with an increased proportion of propionate without affecting feed degradability. In the in vivo experimentation, eighteen female buffalo calves (10-14 months old, avg. body weight 131.68 ± 7.50 kg) were divided into three groups (CONT, EPLM-1, and EPLM-2) of six each in a completely randomized design. Treatment groups were supplemented with a blend (1:1) of dry grounded eucalyptus ( Eucalyptus citriodora ) and poplar ( Populus deltoids ) leaves (50 g, EPLM-1; 150 g, EPLM-2). Feed intake and growth rate of buffalo calves fed on different feeding regimens did not differ ( p > 0.05). Haemato-biochemical parameters reveal no variations ( p > 0.05) among groups, irrespective of period of collection, except the concentration of blood urea, which was decreased ( p < 0.05) in both treatment groups as compared to the control. The levels of reduced glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD) increased ( p < 0.05) in EPLM supplemented animals as compared to CONT. The extent of lipid peroxidation (LPO) was reduced ( p < 0.05) with increased level of EPLM in the diet. The total thiol group (T-SH) was also increased with the supplementation of leaves in the diet. The mean absolute values for skin thickness following the intra-dermal injection of PHA-P were increased ( p < 0.05) in all the supplemented animals relative to CONT. However, there was no significant difference among the calves fed graded levels of these feed additives in the diet. The Ab titer against Pasteurella multocida vaccine was higher ( p < 0.05) on both day 45 and 90 in both treatments, irrespective of the level of additive supplemented. The enteric methane production was reduced in EPLM supplemented buffaloes; however, digestibility of all the nutrients remained comparable ( p > 0.05) among the animals. It may be concluded that blends of eucalyptus ( Eucalyptus citriodora ) and poplar ( Populus deltoides ) leaf-meal (50 g/h/d) containing 3.19 g, 2.30 g, and 0.71 g of total phenolics, tannin phenolics, and condensed tannins, respectively, can be used as the phytogenic feed additive for improving antioxidant status and immunity of buffalo calves, and mitigating enteric methane production without affecting performance and nutrient utilization.

Foot-and-mouth disease (FMD) is endemic in India with a preponderance of outbreaks caused by FMD virus (FMDV) serotype O. Out of the 11 global topotypes of serotype O, only ME-SA topotype has been reported in the country so far. Lineage O/ME-SA/Ind2001 and O/ME-SA/PanAsia are documented as the most dominant ones in terms of the number of outbreaks caused by them. To understand the distribution of topotype/lineages in India and their antigenic behaviour during the year 2014-2018, a total of 286 FMDV serotype O viral isolates were sequence determined at the VP1 region, and 109 isolates were characterized antigenically. All the isolates grouped in the ME-SA topotype, being distributed in lineage O/ME-SA/Ind2001 (within sub-lineages O/ME-SA/Ind2001d and O/ME-SA/Ind2001e), and a new group designated here as O/ME-SA/2018 cluster. The sub-lineage O/ME-SA/Ind2001e reported for the first time in India during the year 2015, replaced sub-lineage O/ME-SA/Ind2001d gradually, which was dominating since 2008. During the years 2014-2018, the sub-lineage O/ME-SA/Ind2001e was found to be the most predominant one whose mean evolutionary rate was observed to be faster than that of the sub-lineage O/ME-SA/Ind2001d. The codon sites 45 and 85 of VP1 were found to be under diversifying selection in a large proportion of trees. The common ancestor predicted for sub-lineages O/ME-SA/Ind2001e and O/ME-SA/2018 dates back to 2012 and 2016, respectively. The sustenance and spread of the new O/ME-SA/2018 cluster need to be assessed by continued surveillance. The Indian vaccine strain O/INDR2/1975 was found to provide adequate antigenic coverage to the emerging and prevalent serotype O lineages. The trait association tests showed frequent virus exchange among different states, which could be an important confounder in the region-specific assessment of effectiveness of FMD control programme., (© 2020 Wiley-VCH GmbH.)

Background: RT-qPCR technique is the current world-wide method used for the early detection of SARS-CoV2 RNA in the suspected clinical samples. Viral RNA extraction is the key pre-analytical step for SARS-CoV2 detection which often achieved using commercial RNA-extraction kits. However, due to the COVID-19 pandemic, bulk production and the supply chains for the commercial RNA-extraction kit have been seriously compromised. The shortage of commercial RNA-extraction kit is even more acute in developing country. Furthermore, use of one-off design RNA-columns can generate plastic wastes that have an environmental pollution effect., Methods and Results: To address these issues, in this study, we used warm alkaline solution containing Triton X-100 for the complete removal of the residual SARS-CoV2 RNA from the used RNA-binding silica column. Columns regenerated using the alkaline solution have the viral RNA purification capability that is comparable to the fresh silica columns. We also demonstrated that RNA-binding silica columns can be regenerated and reused for a minimum of five-times., Conclusions: Therefore, the use of the RNA-column regeneration method may benefits several SARS-CoV2 diagnostic laboratories throughout the world by cutting down the requirement of commercial RNA-purification column., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Several feed additives have been used in recent past to reduce enteric methane production in ruminants. But most of them also inhibit feed digestibility and rumen fermentation, thereby lowering animal performance. Phytogenic feed additives are gaining importance owing to their safety in regard to human health issues. The present study examined the effect of dietary supplementation of phytogenic feed additive containing a blend of poplar (Populus deltoides) and eucalyptus (Eucalyptus citriodora) leaves on feed utilization, milk production, methane emissions and health status of Murrah buffaloes (Bubalus bubalis)., Results: The daily milk yield, 6% fat corrected milk yield, and fat-protein corrected milk yield were increased (P < 0.05) in phytogenic composite feed additive (PCFA)-supplemented buffaloes. A decrease (37.3%) in methane concentration in exhaled air of supplemented buffaloes was evident. The digestibility coefficient of dry matter, organic matter and neutral detergent fibre was increased (P < 0.05) in PCFA-fed buffaloes without affecting feed intake. Total digestible nutrient content of the ration fed to buffaloes of the PCFA group was significantly (P < 0.05) increased. The buffaloes fed PCFA exhibited an enhanced cell-mediated and humoral immune response., Conclusion: A discernible positive impact was evident on overall performances and health status along with lowered methane production of buffaloes fed (15 g kg -1 dry matter intake) a blend of phytogenic feed additive composed of leaves of poplar (P. deltoides) and eucalyptus (E. citriodora). © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Camelpox is a wide-spread infectious viral disease of camelids. An outbreak of camelpox was reported in 15 adult male dromedary camels aged between 10 to 16 years of an organized herd in winter season. The infected camels showed clinical signs of fever, anorexia, lachrymation, pendulous lips, excessive salivation and pock lesions on the skin of head, neck, mouth, lips, extremities, thigh, abdomen, scrotum and inguinal region. Mortalities were recorded in three infected camels after 10-12 days of infection and showed systemic pox lesions characterized by vesicles, papules, ulcerations and raised pock lesions in the mucous membranes of the mouth, tongue, tracheal mucosa, lung, abomasum and liver. Histopathology study revealed characteristic pox lesions with intracytoplasmic eosinophilic inclusion bodies in tongue. Lung showed lesion of interstitial pneumonia ( n = 2) and bronchointerstitial pneumonia ( n = 1). Liver showed infiltration of mononuclear cells around central veins and degenerative changes of hepatocytes. The abomasum and intestine showed ulcerations, marked capillary congestion and areas of lymphocyte infiltration in mucosa and submucosa. The presence of camelpox virus (CMLV) was confirmed in viral DNA isolated from formalin fixed paraffin embedded (FFPE) tissues of tongue, lung, abomasum, liver, heart and intestine of infected camels by C18L gene PCR. The sequencing of viral DNAs showed phylogenetic relatedness with other CMLV isolates from India and other countries. Thus, our study confirmed the rare severe form of systemic camelpox outbreak in adult male dromedary camels hence future attention should be given for studies on virulence, strain identification and molecular epidemiology of CMLV for planning of effective preventive and control strategies., Competing Interests: The authors declare no conflict of interest., (© 2021 The Author(s).)

In the present study, sudden mortalities were reported due to pneumonia in four neonatal camels (5 to 10 days old) of an organized dromedary camel farm. The clinical manifestations in affected camels were weakness, mild to high fever, not suckling, respiratory distress, and sudden death. On the basis of gross and histopathological lesions, the pneumonia was classified into bronchopneumonia (n = 2), bronchointerstitial pneumonia (n = 1), and interstitial pneumonia (n = 1). In bacterial culture, Klebsiella pneumoniae was isolated from lung of all the four cases. These bacteria were also confirmed by PCR from DNA isolated from culture as well as lung tissue. The sequencing of PCR products from four isolates of K. pneumoniae revealed homology with other pathogenic K. pneumoniae isolates from India and other countries. These findings confirmed the role of K. pneumoniae as an important etiological agent for causing pneumonia with sudden mortalities in suckling neonatal dromedary camels.

Hepatic encephalopathy (HE) is a debilitating neuropsychiatric complication associated with acute and chronic liver failure. It is characterized by diverse symptoms with variable severity that includes cognitive and motor deficits. The aim of the study is to assess metabolic alterations in the brain and liver using nuclear magnetic resonance (NMR) spectroscopy and subsequent multivariate analyses to characterize metabolic signatures associated with HE. HE was developed by bile duct ligation (BDL) that resulted in hepatic dysfunctions and cirrhosis as shown by liver function tests. Metabolic profiles from control and BDL rats indicated increased levels of lactate, branched-chain amino acids (BCAAs), glutamate, and choline in the liver, whereas levels of glucose, phenylalanine, and pyridoxine were decreased. In brain, the levels of lactate, acetate, succinate, citrate, and malate were increased, while glucose, creatine, isoleucine, leucine, and proline levels were decreased. Furthermore, neurotransmitters such as glutamate and GABA were increased, whereas choline and myo-inositol were decreased. The alterations in neurotransmitter levels resulted in cognitive and motor defects in BDL rats. A significant correlation was found among alterations in NAA/choline, choline/creatine, and NAA/creatine with behavioral deficits. Thus, the data suggests impairment in metabolic pathways such as the tricarboxylic acid (TCA) cycle, glycolysis, and ketogenesis in the liver and brain of animals with HE. The study highlights that metabolic signatures could be potential markers to monitor HE progression and to assess therapeutic interventions.

The study evaluated the potential of blends of eucalyptus oil and aqueous extract of mulethi (root of Glycyrrhiza glabra ) to reduce rate of ruminal ammonia production without affecting feed digestion to improve nitrogen utilization efficiency and performance of Murrah buffalo ( Bubalus bubalis ). Based on preliminary independent studies with graded doses of eucalyptus oil and mulethi root aqueous extract in modulating in vitro rumen fermentation, four blends of feed additive comprising graded doses (5, 10, 15, and 25 μL) of eucalyptus oil and a fixed quantity (15 μL) of aqueous extract of mulethi roots were prepared and examined for their effects on in vitro rumen fermentation and on methane and gas production in 100-mL calibrated glass syringes by standard IVGP protocol. Rumen liquor was collected from four rumen fistulated Murrah buffaloes fed a total mixed ration. Out of four blends, blend-1 comprising 5 μL of eucalyptus oil and 15 μL of aqueous extract (233.6 g/L DW) of mulethi per 40 mL of in vitro medium was found to reduce ammonia production significantly ( p < 0.001) without affecting feed digestibility. An equivalent dose of blend-1 (10.5 mL of eucalyptus oil and 7.35 g of mulethi root powder/h/day) fed to four rumen fistulated buffaloes for 24 days resulted in 50% reduction ( p < 0.05) in rumen ammonia level with no inhibition in feed fermentation or short-chain fatty acid production. The total bacterial population including Ruminococcus albus, Fibrobacter succinogenes, Butyrivibrio fibrisolvens , and Megasphaera elsdenii as well as anaerobic fungi and methanogenic archaea remained unaffected ( p > 0.05). Twelve buffalo calves (avg. BW 137.5 ± 9.2 kg, 8-12 months old) divided into two groups of six each and fed a total mixed ration (concentrate: roughage; 60:40) with or without supplementation of blend-1 for about 3 months demonstrated 14% increase ( p < 0.05) in average daily gain in BW with a trend ( p < 0.10) in improvement of feed or protein utilization efficiency (1.4 vs. 1.1 g crude protein/g average daily gain; 21.4% increase). Thus, supplementation of eucalyptus oil-mulethi root blend could reduce ruminal ammonia production and improve feed utilization efficiency in ruminants., (Copyright © 2020 Chanu, Paul, Dey and Dahiya.)

Ruminant livestock production processes are the major sources of methane production in agriculture sector triggering global environmental pollution. Above 90% of world buffalo population present in Asian countries, India ranks first and contributes significantly to the environmental pollution by enteric methane emissions. In this study, we examined the effect of dietary composite feed additive supplementation on ruminal methane production, nutrient utilization, milk production and immune status of buffaloes (Bubalus bubalis). Eighteen lactating Murrah (Bubalus bubalis) buffaloes at early stage of lactation were divided into two groups of nine animals and fed a composite feed additive [consisted of (%, w/w) dried and ground leaves of Cordia dichotoma and Holoptelea integrifolia, 31.4 each; garlic oil, 0.6; sodium nitrate, 3.1; magnesium sulphate, 8.4; mustard oil, 12.6 and cottonseed oil, 12.5] which contained an ideal combinations of methane inhibitors, alternate hydrogen sinks and rumen stimulating agents to treatment (CFA) group animals along with basal feed of chaffed green sorghum (Sorghum bicolor) fodder, chaffed wheat straw and concentrate mixture for maintenance and milk production. The results showed a decrease (44.6%) in methane concentration in exhaled air of CFA group buffaloes with increase (p < 0.05) in digestibility of feed in comparison to control (CON). Total digestible nutrient (TDN) content of the ration fed to buffaloes of CFA group was significantly (p < 0.05) increased. The daily milk yield, 6% fat corrected milk (FCM) yield and immune response were also increased (p < 0.05) in CFA group. The study suggests that the supplementation of composite feed additive was effective to reduce enteric methane emissions and improvement in production performance and immune status of buffaloes.

Sources of supplemental minerals in the diet of animals are of important significance. Bio-availability of organic sources is believed to be more in the body as compared to regularly used inorganic sources and hence environment-friendly due to reduced mineral excretion, which in turn reduces their requirements in the diet as well. Twenty-four male Murrah buffalo (Bubalus bubalis) calves (about 18-20 months of age and 318.54 ± 8.85 kg body weight) were divided randomly into four groups of six animals each. In the control group (C, InOrg100) zinc (Zn), copper (Cu), and manganese (Mn) were supplemented through an inorganic source, while in treatment groups, organic source at the rate of 50, 75, or 100% (in groups T1 (Org50), T2 (Org75), and T3 (Org100), respectively) was fed at level as supplemented in the control group. Feeding was continued for a period of 180 days with blood sampling at day 0 followed by a regular interval of 45 days. Plasma samples were analyzed for trace elements Cu, Mn, Zn, and iron (Fe), total antioxidant status, ceruloplasmin, and superoxide dismutase (SOD) with cell-mediated and humoral immune response. Plasma levels of different trace minerals like Fe, Mn, and Cu remained unaffected with two sources and different levels of organic minerals, except the level of Zn, which showed higher (P < 0.05) levels in the group Org100 compared to others, and remained indicative of higher bio-availability through the organic source. The concentration of plasma total antioxidants indicated no adverse effect on the reduction of supplemental levels up to half of these minerals. Also, the level of plasma SOD was high (P < 0.05) at each level of the organic source as compared to the 100% level of the inorganic source. Immune response in respect of cell-mediated as well as humoral immunity did not show any reduction in different groups. The study indicated beneficial impacts of the organic source in the form of superior plasma Zn level as well as SOD concentrations. In addition, no negative effect on most of the studied parameters was observed after reducing supplemental trace minerals to half indicating higher bio-availability of organic trace minerals.

Camelpox is an important viral disease of camels, which may produce mild skin lesions or severe systemic infections. Camelpox virus (CMLV) isolates retrieved from an incidence of camelpox in camels at Bikaner, India were characterized on the basis of genotype and pathotype. Histopathological examination of the CMLV scab revealed intracytoplasmic-eosinophilic inclusion bodies. The phylogenetic analysis of all eight CMLV isolates for C18L gene nucleotide sequence revealed its clustering with its strains M-96 from Kazakhstan and CMS from Iran. The study will help to understand the transmission chain, pathobiology, and epidemiology of circulating CMLV strains. The full genome sequencing of some of the exemplary samples of CMLV is recommended in order to plan and implement a suitable control strategy., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Camelpox caused by a Camelpox virus (CMLV) is a very important host specific viral disease of camel. It is highly contagious in nature and causes serious impact on health even mortality of camels and economic losses to the camel owners. It manifests itself either in the local/mild or generalized/severe form. Various outbreaks of different pathogenicity have been reported from camel dwelling areas of the world. CMLV has been characterized in embryonated chicken eggs with the production of characteristic pock lesions and in various cell lines with the capacity to induce giant cells. Being of Poxviridae family, CMLV employs various strategies to impede host immune system and facilitates its own pathogenesis. Both live and attenuated vaccine has been found effective against CMLV infection. The present review gives a comprehensive overview of camelpox disease with respect to its transmission, epidemiology, virion characteristics, viral life cycle, host interaction and its immune modulation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The present study investigated the pathological features of tuberculosis (TB) caused by Mycobacterium bovis and its diagnosis in naturally infected dromedary camels from an organized farm in India. During the period of the 5-year study, a total of 18 (19.56 %) camels out of 92 examined showed gross lesions compatible with TB at post-mortem. The clinical signs and pathological lesions in these camels were studied, and the efficacy of different diagnostic tests was also assessed. On the basis of occurrence and distribution of gross TB lesions, the infected camels revealed two different lesional patterns as pulmonary (n = 15) and disseminated (n = 3) form. The histopathology of affected organs revealed typical granulomatous lesions wherein the giant cells and acid-fast bacilli were occasionally observed in pulmonary form whereas they frequently observed in disseminated form. The single intradermal tuberculin test (SIDT) detected TB in 10 (55.55 %) whereas the Ziehl-Neelsen (ZN) stain and IS6110 PCR from tissue lesions detected 13 (72.22 %) and 18 (100 %) of the infected camels, respectively. The study suggests that pulmonary form of the TB is more common in camels indicating respiratory route as the major source of exposure in camel herds. Moreover, very low sensitivity of SIDT was observed which highlights the difficulty for confirmation of TB in live camels.

The present study describes the PCR amplification of GM-CSF-inhibitory factor (GIF) and Uracil DNA glycosylase (UDG) encoding genes of pseudocowpoxvirus (PCPV) from the Indian Dromedaries (Camelus dromedarius) infected with contagious ecthyma using the primers based on the corresponding gene sequences of human PCPV and reindeer PCPV, respectively. The length of GIF gene of PCPV obtained from camel is 795 bp and due to the addition of one cytosine residue at position 374 and one adenine residue at position 516, the open reading frame (ORF) got altered, resulting in the production of truncated polypeptide. The ORF of UDG encoding gene of camel PCPV is 696 bp encoding a polypeptide of 26.0 kDa. Comparison of amino acid sequence homologies of GIF and UDG of camel PCPV revealed that the camel PCPV is closer to ORFV and PCPV (reference stains of both human and reindeer), respectively., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The Toll-like receptor 2 (TLR2) gene of old world camels (Camelus dromedarius and Camelus bactrianus) was cloned and sequenced. The TLR2 gene of the dromedary camel had the highest nucleotide and amino acid identity with pig, i.e., 66.8% and 59.6%, respectively. Similarly, the TLR2 gene of the Bactrian camel also had the highest nucleotide and amino acid identity with pig, i.e., 85.7% and 81.4%, respectively. Dromedary and Bactrian camels shared 77.9% nucleotide and 73.6% amino acid identity with each other. Interestingly, the amidation motif is present in camel (Dromedary and Bactrian) TLR2 only, and the TIR domain is absent in Dromedary camel TLR2. This is the first report of the TLR2 gene sequence of Dromedary and Bactrian camels.

The dsRNA binding protein (RBP) encoding gene of parapoxviruses (PPVs) from the Dromedary camels, inhabitating different geographical region of Rajasthan, India were amplified by polymerase chain reaction using the primers of pseudocowpoxvirus (PCPV) from Finnish reindeer and cloned into pGEM-T for sequence analysis. Analysis of RBP encoding gene revealed that PPV DNA from Bikaner shared 98.3% and 76.6% sequence identity at the amino acid level, with Pali and Udaipur PPV DNA, respectively. Reference strains of Bovine papular stomatitis virus (BPSV) and PCPV (reindeer PCPV and human PCPV) shared 52.8% and 86.9% amino acid identity with RBP gene of camel PPVs from Bikaner, respectively. But different strains of orf virus (ORFV) from different geographical areas of the world shared 69.5-71.7% amino acid identity with RBP gene of camel PPVs from Bikaner. These findings indicate that the camel PPVs described are closely related to bovine PPV (PCPV) in comparison to caprine and ovine PPV (ORFV).

The antiviral potential of small interfering RNAs (siRNAs) targeting rabies virus (RV) polymerase (L) and nucleoprotein (N) genes delivered through lentiviral vector was investigated. For in vitro evaluation, siRNAs expressing BHK-21 cell lines (BHK-L and BHK-N) were developed using transduction with Lenti-L and Lenti-N lentiviruses encoding siRNAs against RV-L and N genes, respectively. When these cell lines were challenged in vitro with RV Pasteur virus-11 (PV-11) strain, there was reduction in number of RV-specific foci and target gene transcripts indicating inhibitory effect on RV multiplication. For in vivo evaluation, mice were treated intracerebrally with lentiviruses and challenged with 20 LD50 of RV challenge virus standard-11 (CVS-11) strain by intramuscular route in masseter muscle. Five out of eight mice treated with Lenti-N survived indicating 62.5 % protection. The control and Lenti-L-treated mice died within 7-10 days indicating lethal nature of challenge virus and no protection. These results demonstrated that siRNA targeting RV-N could not only inhibit RV multiplication, but also conferred protection in mice against lethal RV challenge. These findings have implication on therapeutic use of siRNA targeting RV-N against RV infection.

The haemagglutinin (HA) encoding gene and genes encoding for immunomodulatory proteins i.e., schlafen-like protein, epidermal growth factor and golgi anti apoptotic protein of camelpoxvirus (CMLV) obtained from Indian dromedarian camels were cloned and characterized. In this study, the size of the HA encoding gene obtained from the Indian CMLV is 941 bp which is only partial. Sequence analysis of schlafen-like protein gene revealed that CMLV obtained from India shared 99.6% identity with CMLV-Iran and CMLV-Kazakhstan strains both at nucleotide and amino acid level. The size of epidermal growth factor (EGF) gene of Indian CMLV obtained in this study was 418 bp, which was due to the addition of one cytosine residue position 132 of EGF gene of Indian CMLV. Sequence analysis revealed that the Golgi anti-apoptotic protein (GAAP) of Indian CMLV shared 99.5% sequence identity both at the nucleotide and amino acid level with CMLV-Kazakhstan. Based on the nucleotide and amino acid sequence identities and phylogenetic analyses of these genes, it is found that CMLV-India is forming a cluster with Kazakhstan and Iranian CMLV isolates., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Cellular interleukin-10 (IL-10) gene from the peripheral blood mononuclear cells of the healthy Dromedary camel (Camelus dromedarius) and viral IL-10 (vIL-10) from the skin scabs of the Dromedary camels infected with contagious ecthyma (a parapoxviral infection in the camels) were amplified by polymerase chain reaction, cloned and characterized. Sequence analysis revealed that the open reading frame (ORF) of dromedarian camel IL-10 is 537 bp in length, encoding 178 amino acid polypeptide while open reading frame of vIL-10 from camel is 561 bp, encoding 187 amino acid polypeptide. The Dromedary camel IL-10 exhibited 62.6% and 68.5% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from camel. Sequence analysis also revealed that the Dromedary camel IL-10 shared 99.4% and 98.3% identity at the nucleotide and amino acid level, respectively, with the Bactrian camel (Camelus bactrianus). But vIL-10 from camel shared 84.7% and 83.4% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from reindeer (Rangifer tarandus), which is a ruminant species belonging to the order Artiodactyla. The present study was conducted to evaluate the evolutionary origin of the camel parapoxvirus with parapoxviruses of cattle and sheep and the resultant sequence analysis revealed that camel parapoxvirus is closely related to cattle parapoxvirus than sheep parapoxvirus (Orf virus)., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A replicon-based DNA vaccine encoding VP2 gene of canine parvovirus (CPV) was developed by cloning CPV-VP2 gene into a replicon-based DNA vaccine vector (pAlpha). The characteristics of a replicon-based DNA vaccine like, self-amplification of transcripts and induction of apoptosis were analyzed in transfected mammalian cells. When the pAlpha-CPV-VP2 was injected intradermal as DNA-launched replicon-based DNA vaccine in dogs, it induced CPV-specific humoral and cell mediated immune responses. The virus neutralization antibody and lymphocyte proliferative responses were higher than conventional CPV DNA vaccine and commercial CPV vaccine. These results indicated that DNA-launched replicon-based CPV DNA vaccine was effective in inducing both CPV-specific humoral and cellular immune responses and can be considered as effective alternative to conventional CPV DNA vaccine and commercial CPV vaccine., (Crown Copyright © 2012. Published by Elsevier India Pvt Ltd. All rights reserved.)

To evaluate antiviral potential of adenoviral vector-delivered small interfering RNA (siRNA) against rabies, recombinant, replication-defective adenoviral vectors (rAdV) encoding siRNAs targeting rabies virus (RV) polymerase (L) and nucleoprotein (N) genes were developed. The siRNAs were delivered as small hairpin RNAs (shRNAs) through these vectors. Treatment of BHK-21 cells with rAdV expressing siRNA targeting L gene (rAdV-L) and N gene (rAdV-N) (100 MOI) and their subsequent infection with RV (0.001 MOI, RV PV-11), reduced RV fluorescent foci by 48.2% (mean±SEM; 48.17±0.6540, N=6) and 41.8% (mean±SEM; 41.83±0.3073, N=6), respectively, with respect to that of BHK-21 cells treated with rAdV expressing negative control siRNA (rAdV-Neg) indicating inhibition of multiplication of RV in BHK-21 cells in response to adenoviral vector mediated siRNA delivery. Also, the similar treatment of BHK-21 cells with rAdV-L and rAdV-N and similar subsequent infection of them with RV resulted in reduction in RV mRNA transcript levels for their respective targets (RV L gene for rAdV-L and N gene for rAdV-N). mRNA transcript level for RV L gene was reduced by 17.88-fold (mean±SEM; 17.88±0.06638, N=6) in cells treated with rAdV-L and that for RV N gene was reduced by 5.7-fold (mean±SEM; 5.7±0.04472, N=6), in cells treated with rAdV-N, in comparison with that in cells treated with rAdV-Neg, as analyzed by using real-time PCR. These in vitro studies showed that between these two, adenoviral vector mediated delivery of siRNA targeting RV L gene was comparatively more effective in inhibiting RV multiplication in BHK-21 cells than that of siRNA targeting RV N gene (p<0.0001). Localized treatment (intramuscular injection in masseter muscle) of mice with 10(7) plaque forming units of either rAdV-L or rAdV-N and subsequent lethal RV infection (15-20LD(50) of CVS-11) at the same site, through the same route, although resulted in 50% protection (3 out of 6 mice survived) against lethal rabies, the survival patterns for groups of mice treated with either rAdV-L or rAdV-N and that treated with rAdV-Neg did not differ significantly (p=0.5234). These results indicated that adenoviral vector mediated siRNA delivery, in vitro in BHK-21 cells inhibited RV multiplication in vitro in BHK-21 cells; siRNA targeting RV L gene used in this study was comparatively more efficient in doing this than that targeting RV N gene used in this study; in vivo in mice inhibited RV multiplication in mice and imparted partial protection against lethal rabies and so it may have a potential to be used as an alternative antiviral approach against rabies, although further study is required to establish its efficacy for this purpose., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

To investigate the potential of RNA interference (RNAi) as antiviral agent against rabies, two small interfering RNAs (siRNAs) targeting rabies virus (RABV) nucleoprotein (N) and polymerase (L) genes were designed and evaluated. Both siRNAs knockdown or silenced the target RABV genes as evaluated in a plasmid based transient expression model. For efficient delivery, adenoviruses expressing the siRNAs were constructed and antiviral potential of the delivered siRNAs was investigated in BHK-21 cells. When cells treated with adenoviruses expressing siRNAs were challenged with RABV, there was 88.35±2.4% and 41.52±9.3% reduction in RABV multiplication in infected cells with siRNAs targeting RABV-N and L genes, respectively. Relative quantification of RABV transcripts using real-time PCR revealed knockdown of both RABV-N and L gene transcripts, however, significant reduction was observed only with adenovirus expressing siRNA against RABV-N. When mice treated intracerebrally with adenoviruses expressing siRNAs were challenged peripherally with lethal RABV by the intramuscular route in masseter muscle, there was 66.6% and 33.3% protection with adenoviruses expressing siRNAs against RABV-N and L genes, respectively. These results demonstrated that adenovirus expressing siRNA against RABV-N efficiently inhibited the RABV multiplication both, in vitro and in vivo and conferred significant protection against lethal RABV challenge. This supported the hypothesis that RNAi, based on siRNA targeting RABV-N gene can prevent RABV infection and holds the potential of RNAi as an approach to prevent rabies infection., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Topoisomerase gene of pseudocowposvirus from Indian dromedarian camel was amplified by PCR using the primers of PCPV from Finnish reindeer and cloned into pGEM-T for sequence analysis. Analysis of amino acid identity revealed that Indian PCPV of camel shared 95.9-96.8 with PCPV of reindeer, 96.2-96.5 with ORFV and 87.5 with BPSV., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The complete genome of a lapinized classical swine fever virus (CSFV) vaccine strain was amplified into nine overlapping fragments by RT-PCR, and nucleotide sequences were determined. Complete genome sequence alignment and phylogenetic analysis indicated 92.6-98.6% identities at the nucleotide level with other reported CSFV strains and could be grouped into subgroup 1.1 along with other attenuated strains of CSFV. The 5'-UTR demonstrated >97.0% nucleotide similarity with most of vaccine CSFV strains from China. Further, its 3'-UTR sequence indicated a length similar to all the CSFV strains from China with >98.0% nucleotide similarity, although high length heterogeneity of 3'-UTR was reported among different CSFV strains. There was 12 nt (TTTTCTTTTTTT) insertion in 3'-UTR similar to other reported attenuated vaccine strains. However, secondary structure of 3'-UTR indicated that Indian CSFV strain requires further passage to obtain a 3'-UTR structure similar to most of the attenuated strains.

A Sindbis virus replicon-based DNA vaccine containing VP2 gene of canine parvovirus (CPV) was delivered by Escherichia coli to elicit immune responses. The orally immunized dogs developed CPV-specific serum IgG and virus neutralizing antibody responses. The cellular immune responses analyzed using lymphocyte proliferation test and flow cytometry indicated CPV-specific sensitization of both CD3+CD4+ and CD3+CD8+ lymphocytes. This study demonstrated that the oral CPV DNA vaccine delivered by E. coli can be considered as a promising approach for vaccination of dogs against CPV.

A self-replicating RNA vaccine encoding rabies virus glycoprotein gene was developed utilizing sindbis virus RNA replicon. The in vitro transcribed RNA (Sin-Rab-G RNA) was transfected in mammalian cells and analysed for self-replication and expression of rabies glycoprotein. To generate immune responses against rabies, mice were immunized with 10microg of Sin-Rab-G RNA and immune responses developed were compared with mice immunized with rabies DNA vaccine and commercial cell culture vaccine (Rabipur). The self-replicating rabies RNA vaccine generated cellular and humoral IgG responses similar to rabies DNA vaccine. On challenge with rabies virus CVS strain, rabies RNA vaccine conferred protection similar to rabies DNA vaccine. These results demonstrated that replicon-based self-replicating rabies RNA vaccine with 10microg dose was effective in inducing immune responses and protection similar to rabies DNA vaccine.

A Sindbis virus (SINV) replicon-based DNA vaccine encoding Rabies virus (RABV) glycoprotein G developed previously (Saxena et al., Vaccine 26, 6592, 2008) was used for immunization of dogs against rabies. The intradermal injection of DNA vaccine into external ear generated protective level of virus neutralizing antibodies. The cellular immune response was specific to RABV, in particular by an increase in CD3+CD4+ and CD3+CD8+ lymphocytes. This study has demonstrated that the SINV replicon-based DNA vaccine encoding RABV G is capable of inducing the protective level of specific immune response in dogs.

A sindbis virus replicon-based DNA vaccine encoding rabies virus glycoprotein (G) was developed by subcloning rabies G gene into a sindbis virus replicon-based vaccine vector (pAlpha). The self-amplification of RNA transcripts and translation efficiency of rabies G was analyzed in pAlpha-Rab-G-transfected mammalian cells using RT-PCR, SDS-PAGE and Western blot analysis. The transfected cells also showed induction of apoptosis which is an important event in the enhancement of immune responses. Further, immune responses induced with replicon-based rabies DNA vaccine (pAlpha-Rab-G) was compared with conventional rabies DNA vaccine and commercial cell culture vaccine (Rabipur) in intramuscularly injected mice. The mice immunized with replicon-based rabies DNA vaccine induced humoral and cell mediated immune responses better than conventional rabies DNA vaccine however, comparable to Rabipur vaccine. On challenge with rabies virus CVS strain, replicon-based rabies DNA vaccine conferred complete protection similar to Rabipur. These results demonstrate that replicon-based rabies DNA vaccine is effective in inducing both humoral and cellular immune responses and can be considered as effective vaccine against rabies.

Adverse alterations in lipid profile suggesting higher atherogenicity were observed following 12 weeks treatment with atenolol in patients of hypertension. No significant alterations in lipid profile were observed with labetalol therapy.