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Respiratory tract infections are of serious concern to the poultry industry. The present study was aimed to delineate the extent of respiratory avian mycoplasmosis associated bacterial and viral concurrent infections in the poultry flocks. A total of 146 poultry flocks of Haryana and Rajasthan, India, suspected for chronic respiratory disease (CRD) were screened for avian mycoplasmas, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and avian pathogenic Escherichia coli (APEC) by conventional polymerase chain reaction (PCR) assays. A total of 49.31% (72/146) flocks were found positive for Mycoplasma infection. Of the Mycoplasma-positive flocks, 80.55% (58/72) represented pathogenic avian mycoplasmas (MG and/or MS), while 19.44% (14/72) flocks were positive for commensal avian mycoplasmas (other than MG and MS). A correlation was deduced between avian mycoplasmosis and bacterial and/or viral co-infections. The results revealed that 17.24% (10/58) flocks had only avian mycoplasmosis infection. However, in the remaining flocks, the avian mycoplasmosis was associated either with APEC infection [17.24% (10/58)], IBV infection [43.10% (25/58)], or both APEC and IBV infections [22.41% (13/58)], respectively. Further epidemiological studies on respiratory avian mycoplasmosis associated concurrent infections with other pathogens are recommended to assess circulating strains, risk factors, and economic losses.

Newcastle disease virus (NDV) can infect approximately 250 avian species and causes highly contagious Newcastle disease (ND) in domestic poultry, leading to huge economic losses. There are three different pathotypes of NDV, i.e., lentogenic, mesogenic, and velogenic. Wild resident (wild) and migratory birds are natural reservoirs of NDV and are believed to play a key role in transmitting the virus to domestic poultry. The present study was conducted to determine the prevalence of NDV in wild and migratory birds in the state of Haryana, India, during two migratory seasons (2018-19 and 2019-20). In total 1379 samples (1368 choanal swabs and 11 tissue samples) were collected from live ( n = 1368) or dead birds ( n = 4) belonging to 53 different avian species. These samples belonged to apparently healthy ( n = 1338), sick ( n = 30), and dead ( n = 4) birds. All samples were tested for NDV by real-time reverse transcription-PCR using M gene specific primers and probe. Of the 1379 samples, 23 samples from wild birds [ Columba livia domestica ( n = 12, 52.17%), Pavo cristatus ( n = 9, 39.13%), and Psittaciformes ( n = 2, 8.69%)] were found positive for NDV. Only one of the 23 samples (from P. cristatus ) was positive for F gene, indicating it to be a mesogenic/velogenic strain. These results indicate that both lentogenic and velogenic strains of NDV are circulating in wild birds in Haryana and that further studies are needed to characterize NDV strains from wild/migratory birds and domestic poultry to determine the extent of virus transmission among these populations. This study considers the disease transmission risk from domestic pigeons and parrots to commercial poultry and vice versa, and the results emphasize the need for strict biosecurity strategies to protect commercial poultry in the region.

Background: Newcastle disease (ND) is an economically important viral disease affecting the poultry industry. In Kerala, a state in South India, incidences of ND in commercial and backyard poultry have been reported. But a systematic statewide study on the prevalence of the disease has not been carried out., Objectives: A cross-sectional survey was performed to detect the presence of Newcastle disease virus (NDV) in suspect cases and among apparently healthy commercial flocks and backyard poultry, in the state and to identify risk factors for NDV infection., Methods: Real-time reverse transcription-PCR (RT-PCR) was used to detect the M gene of NDV in choanal swabs and tissue samples collected from live and dead birds, respectively and the results were statistically analysed., Results: The predominant clinical signs of the examined birds included mild respiratory signs, huddling together and greenish diarrhoea. Nervous signs in the form of torticollis were noticed in birds in some of the affected flocks. On necropsy, many birds had haemorrhages in the proventriculus and caecal tonsils which were suggestive of ND. Of the 2079 samples tested, 167 (8.0%) were positive for the NDV M-gene by RT-PCR. Among 893 samples collected from diseased flocks, 129 (14.5%), were positive for M gene with pairwise relative risk (RR) of 15.6 as compared to apparently healthy flocks where 6 out of 650 (0.9%) samples were positive. All positive samples were from poultry; none of the ducks, pigeons, turkey and wild birds were positive. Commercial broilers were at higher risk of infection than commercial layers (RR: 4.5) and backyard poultry (RR: 4.9). Similarly, birds reared under intensive housing conditions were at a higher risk of being infected as compared to those reared under semi-intensive (RR: 6.7) or backyard housing (RR: 2.1). Multivariable analysis indicated that significantly higher risk of infection exists during migratory season and during ND outbreaks occurring nearby. Further, lower risk was observed with flock vaccination and backyard or semi-intensive housing when compared to intensive housing. When the M gene positive samples were tested by RT-PCR to determine whether the detected NDV were mesogenic/velogenic, 7 (4.2%) were positive., Conclusions: In Kerala, NDV is endemic in poultry with birds reared commercially under intensive rearing systems being affected the most. The outcome of this study also provides a link between epidemiologic knowledge and the development of successful disease control measures. Statistical analysis suggests that wild bird migration season and presence of migratory birds influences the prevalence of the virus in the State. Further studies are needed to genotype and sub-genotype the detected viruses and to generate baseline data on the prevalence of NDV strains, design better detection strategies, and determine patterns of NDV transmission across domestic poultry and wild bird populations in Kerala., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Newcastle disease (ND), caused by Newcastle disease virus (NDV), is a contagious disease that affects a variety of domestic and wild avian species. Though ND is vaccine-preventable, it is a persistent threat to poultry industry across the globe. The disease represents a leading cause of morbidity and mortality in chickens. To better understand the epidemiology of NDV among commercial and backyard chickens of Odisha, where chicken farming is being prioritized to assist with poverty alleviation, a cross-sectional study was conducted in two distinct seasons during 2018. Choanal swabs (n = 1361) from live birds (commercial layers, broilers, and backyard chicken) and tracheal tissues from dead birds (n = 10) were collected and tested by real-time reverse transcription polymerase chain reaction (RT-PCR) for the presence of matrix (M) and fusion (F) genes of NDV. Risk factors at the flock and individual bird levels (health status, ND vaccination status, geographical zone, management system, and housing) were assessed using multivariable logistic regression analyses. Of the 1371 samples tested, 160 were positive for M gene amplification indicating an overall apparent prevalence of 11.7% (95% CI 10.1-13.5%). Circulation of virulent NDV strains was also evident with apparent prevalence of 8.1% (13/160; 95% CI: 4.8-13.4%). In addition, commercial birds had significantly higher odds (75%) of being infected with NDV as compared to backyard poultry (p = 0.01). This study helps fill a knowledge gap in the prevalence and distribution of NDV in apparently healthy birds in eastern India, and provides a framework for future longitudinal research of NDV risk and mitigation in targeted geographies-a step forward for effective control of ND in Odisha., Competing Interests: The authors have declared that no competing interests exist.

Brucella abortus vaccines play a central role in bovine brucellosis control with tremendous success worldwide for decades. The study was aimed to evaluate the efficacy of reduced dose (5.0 × 10 9 cfu) of S19 vaccine in adult cattle and its shedding in the milk of vaccinated cattle using molecular techniques. The OIE recommended tests (RBPT, SAT, and iELISA) for brucellosis screening in cattle were used. Seronegative cattle (n = 90) of different age groups (young, old heifers & milking cows, n = 30 each) were selected for the vaccine trials. Antibody titers were recorded at 7th, 21st, 30th, 60th, 90th and 120th days post-vaccination (DPV) to monitor the immune responses following vaccination and at 150th, 180th, 210th and 240th DPB following booster-dose to an intraocular group. The humoral immune responses observed by RBPT and ELISA, proved that antibody titers persisted in s/c group compared to the i/o group in all categories. The IFN-γ stimulation (CMI) due to reduced dose vaccination was noticed early as 30th in all groups and declined after 90th DPV, with higher IFN-γ stimulation among the s/c group. The Bcsp31 and IS711 targeted PCR detected the presence of Brucella DNA in milk samples (n = 120) from the vaccinated cows (n = 30) and confirmed by qPCR (TaqMan assay) at 30th, 60th, 90th and 120th DPV. A Significant number, 70% (7/10) was detected in s/c by qPCR. BCSP31 sequence was deposited at NCBI GenBank (accession no. MK881173-6). PCR and qPCR techniques could provide a reliable diagnosis of brucellosis from milk. The intraocular route remains the safer route for vaccinating adult cattle than subcutaneous., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

Avian mycoplasmosis, mainly caused by Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), is an economically important disease of poultry industry. The present study was aimed to develop duplex PCR as a rapid, specific and economical method for accurate detection of MG and MS in poultry and its comparison with single (monoplex) MG/MS PCR. During present investigation, a total of 146 poultry flocks having clinical history of respiratory disease were screened. Pooled tissue samples (trachea, lungs and air sacs) from 4-5 birds of each flock were collected during necropsy at disease investigation laboratories, Hisar, Haryana, India. The single and duplex PCR assays were standardized using primers of intergenic spacer region (IGSR; 16S-23S rRNA) for MG and hemagglutinin vlhA gene for MS, with expected amplicon size of 812 bp and 1200 bp products, respectively. In single PCR, 6.85%, 2.74% and 2.74% tissue samples were found positive for MG, MS and both MG and MS, respectively. However, duplex PCR showed, 7.53%, 2.74% and 1.37% positivity for MG, MS and both MG and MS, respectively. Taking the results of monoplex PCR as a gold standard, sensitivity and specificity of the developed duplex PCR was found to be 94.44% and 100%, respectively. Moreover, Cohen's kappa statistic (k = 0.97) measured a 'perfect' agreement between monoplex and duplex PCR assays. The positive and negative predictive values of duplex PCR was found to be 1.0 and 0.9922, respectively at 95% confidence interval (CI), as compared to monoplex PCR. The simultaneous use of two genes in a duplex PCR was more rapid and economical than two separate single PCR reactions., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Newcastle disease virus (NDV) causes Newcastle disease (ND) in poultry. The ND is a highly contagious disease, which is endemic in several countries despite regular vaccination with live or killed vaccines. Studies on NDV in India are mostly targeted toward its detection and characterization from disease outbreaks. A surveillance study was undertaken to determine NDV prevalence throughout the state of Haryana from March 2018 to March 2020 using a stratified sampling scheme. The state was divided into three different zones and a total of 4,001 choanal swab samples were collected from backyard poultry, commercial broilers, and layers. These samples were tested for the M gene of NDV using real-time RT-PCR. Of the 4,001 samples tested, 392 were positive (9.8% apparent prevalence; 95% CI: 8.9-10.8%) for the M gene. Of these 392 M gene positive samples, 35 (8.9%; 95% CI: 6.4-12.3%) were found to be positive based on F gene real-time RT-PCR. Circulation of NDV in commercial and backyard poultry highlights the importance of surveillance studies even in apparently healthy flocks. The information generated in this study should contribute to better understanding of NDV epidemiology in India and may help formulate appropriate disease control strategies for commercial and backyard birds., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Joshi, Chaudhary, Bansal, Singh, Maan, Mahajan, Ravishankar, Sahoo, Mor, Radzio-Basu, Herzog, Kapur, Goel, Jindal and Goyal.)

Brucellosis caused by facultative intracellular bacteria, Brucella, remains a global threat to both animal and human health. In this study we aimed to identify potential risk factors of bovine brucellosis and to assess the knowledge, attitudes, and practices (KAPs) of livestock keepers in Hisar, India. A standardized questionnaire was used to collate information regarding potential risk factors of bovine brucellosis and livestock owners' KAPs. A total of 127 livestock keepers were involved. Serum samples from their animals (n = 635) were tested for the presence of antibodies against Brucella by Rose Bengal Plate Test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). Out of these, 78 (61.4%) of the herds had at least one seropositive animal, and 302 (47.6%) of the cattle were seropositive. Univariate and multivariate analysis revealed significant associations between intensive farm type (OR = 4.6; 95% CI, 1.6-16.7; P = 0.009), hygienic disposal of aborted fetuses (OR = 0.3; 95% CI, 0.08-0.9; P = 0.04) and herd seropositivity for brucellosis. The majority, 96 (75.6%) of the respondents, were males aged 18-50, and 82 (64.6%) owned a small-backyard farm. Only 51 (40.2%) of the participants knew about brucellosis; out of them, 54.9% (28/51) could not identify clinical signs of brucellosis. Six (11.8%) participants indicated abortion as the most noticeable clinical sign, and 45.1% indicated that consumption of raw milk is associated with high risk of contracting brucellosis. A large proportion of respondents confirmed that milk from their animals was regularly consumed (86.6%) and sold (59.8%) to other people. These results suggest that bovine brucellosis is endemic in Haryana, where Brucella-contaminated milk is likely being regularly sold. Brucellosis control efforts in Haryana should include education programs to raise awareness of the disease and means to control it in cattle and to prevent zoonotic transmission., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The present study describes the development of a novel affordable and rapid visual dot-blot assay using synthetic multiple antigenic peptides (MAP) for simultaneous detection of antibodies to infectious bronchitis virus (IBV) and Newcastle disease virus (NDV). Antibody detection efficiencies of MAP peptides namely, NP1 MAP (Nucleoprotein IBV) and HN MAP (Haemagglutinin-neuraminidase NDV) were studied in solid-phase indirect peptide ELISA. In comparison with the commercial kit, the NP1 MAP showed 89.20% diagnostic sensitivity (DSn) and 85.90% diagnostic specificity (DSp) at 19.45% ROC cut-off. Similarly, HN MAP was evaluated and showed 89.70% DSn and 92.90% DSp at 19.90 % ROC cut-off. The peptides after evaluating their ELISA performance were further used to device a flow-through dot-blot assay (FT-DBA) for simultaneous detection of IBV and NDV antibodies. The kappa value for IBV by FT-DBA in comparison to commercial ELISA was 0.64 whereas for NDV, FT-DBA gave a kappa value of 0.68 in comparison to commercial ELISA indicating substantial agreement between the assays. In essence, the divergent MAP based diagnostic design could provide an alternative for antibody detection of IBV and NDV. Further, the FT-DBA approach could be used for low cost, rapid and pen-side detection of IBV and NDV antibodies simultaneously., (Copyright © 2021 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)

The objective of the study was to develop a bio-safe synthetic peptide ELISA for the detection of antibodies against the infectious bronchitis virus (IBV) using a novel multiple antigenic peptide approach (MAP). After initial ELISA optimization, diagnostic sensitivity (DSn) and specificity (DSp) for the linear peptides were determined using receiver operator curve (ROC) analysis. The peptide IBVP1 showed 90.44% DSn and 88.64% DSp at ROC cut off 22.8% while IBVP2 showed 88.24% DSn and 85.23% DSp at ROC cut off 23.05%. The multimerization of linear peptides to MAP design resulted in the improvement of the diagnostic efficiency up to 94.85% DSn and 92.05% DSp for IBVM1 with 19.95% cut off. A similar improvement in the performance was also observed with 92.65% DSn and 90.91% DSp for IBVM2 at 20.72% cut off. All the peptides were tested for diagnostic specificity and did not show the cross-reactivity with Newcastle disease virus and infectious bursal disease virus positive serum samples. In addition, repeatability testing for all linear and multimeric peptide showed that the coefficient of variation for intra-assay was within the expected limits, ranging from 2.4 to 10.4% and inter-assay coefficient of variation was ranging from 5.56 to 14.3%. In a nutshell, the present study used predicted B cell epitope, the synthetic peptide in linear and multimeric design for IBV antibody detection. The study also highlights peptide antigen with modified scaffold design could be a safe alternative to whole virion-based ELISA for IBV antibody detection., Competing Interests: Conflict of interestOn behalf of all authors, the corresponding author states that there is no conflict of interest., (© King Abdulaziz City for Science and Technology 2020.)

The molecular characterization of three Newcastle disease viruses (NDV) isolated from backyard chickens in the state of Haryana, India, was undertaken. Two genotype II strains and one genotype XIIIc class II isolate with genome sizes of 15,186 and 15,192 nucleotides (nt), respectively, were identified., (Copyright © 2019 Maan et al.)

Zebrafish is emerging as a promising model for the study of human cancers. Several xenograft models of zebrafish have been developed, particularly in larval stages (<48 h post fertilization) when the immune system of fish is not developed. However, xenografting in adult zebrafish requires laborious and transient methods of immune suppression (γ- irradiation or dexamethasone) that limits engraftment and survival of the tumor or fail to recapitulate specific characteristics of malignancies. Thus, the availability of a simple protocol to successfully engraft adult zebrafish, remains a challenge. The current study addresses this limitation and describes a robust method of xenografting in adult zebrafish. We describe a protocol that involves pre-conditioning of Casper, a pigmentation mutant of zebrafish with busulfan that led to a higher rate of engraftment of hepatocellular carcinoma and acute myeloid leukemia cells. To further ascertain the homing characteristics of the injected cancer cells, we transplanted adult zebrafish by two routes of administration and then studied their compartmentalization. This model presents a valuable alternative to rodents to study the biology of these cancers and also a cost-effective platform for evaluation of potential anti-cancer agents., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Aim: The present study was conducted with the following aims: (i) To screen the Salmonella spp. isolates recovered from suspected cases of fowl typhoid for carriage of Class 1 integrons and analyze their association with antimicrobial resistance and (ii) to carry out molecular characterization and phylogenetic analysis of Class 1 integron-integrase ( intI1 ) gene., Materials and Methods: A total of 43 Salmonella isolates were subjected to polymerase chain reaction (PCR) assay to determine the presence of Class1 intI1 . Differences between different serotypes in relation to their carriage of integrons and the differences between strains containing or not containing an integron and being resistant to different antimicrobials were analyzed by Fisher exact test using STATA™ (StataCorp, College Station, TX). Phylogenetic analysis was carried out using MEGA6 software., Results: Out of 43 isolates, 40 (93.02%) were found positive for Class 1 integrons. 35/40 (87.5%) intI1 -positive isolates were multidrug resistance (MDR) (resistant to ≥4 antibiotics), which support the hypothesis of an association between the presence of Class 1 integrons and emerging MDR in Salmonella . There was no significant difference among isolates resistant to different antimicrobials in Class 1 integron carrying isolates and the Class 1 integron negative isolates (p<0.05). Further, there was no significant difference among different serotypes in respect of their carriage of Class 1 integrons., Conclusion: It can be concluded that the high prevalence of Class 1 integrons indicates a high potential of Salmonella isolates for horizontal transmission of antimicrobial genes, especially among Gram-negative organisms.

Bluetongue (BT) and peste-des-petits-ruminants (PPR) are major transboundary diseases of small ruminant, which are endemic in India. Testing of bluetongue virus (BTV) and peste-des-petits-ruminants virus (PPRV) from recent outbreaks (2015-2016) in different regions of Haryana State of India revealed that 27.5% of the samples showed the presence of dual infection of BTV and PPRV. Analysis of Seg-2 of BTV (the serotype-determining protein) showed the presence of BTV-12w in several isolates. However, analysis of N gene fragment amplicons showed that viruses belong to lineage IV were most closely related to a pathogenic strain of PPRV from Delhi. This is the first report of co-circulation of PPRV lineage IV and bluetongue virus serotype 12 in the state., (© 2017 Blackwell Verlag GmbH.)

Swinepox virus (SWPV), a member of the genus Suipoxvirus causes generalized pock-like lesions on the body of domestic and wild pigs. Although outbreak has been reported in India since 1987, virus isolation and genetic characterization remained elusive. In September 2013, an outbreak of acute skin infection occurred in piglets in a commercial piggery unit at Rohtak district in Haryana, India. The presence of SWPV in scab samples collected from piglets succumbed to infection was confirmed by virus isolation, PCR amplification of SWPV-specific gene segments and nucleotide sequencing. Phylogenetic analysis of host-range genes of the SWPV revealed that the Indian isolate is genetically closely related to reference isolate SWPV/pig/U.S.A/1999/Nebraska. To the best of our knowledge this is the first report on isolation and genetic characterization of SWPV from pigs in India., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Aim: The present study was investigated to ascertain the epidemiological status of fowl typhoid (FT) in broilers in some parts of Haryana during January 2011 to December 2013., Materials and Methods: To elucidate the epidemiological status of FT in broiler chickens for the 3 years (2011-2013) and to study the prevalence of various Salmonella serovars in poultry on the basis of culture characteristics, biochemical features, serotyping, and their antibiogram profile from some parts of Haryana (India)., Results: A total of 309 outbreaks of FT were recorded in chickens during this period. Overall percent morbidity, mortality, case-fatality rate (CFR) in broiler chicks due to FT during this period was 9.45, 6.77, and 71.55. The yearly observations were divided into quarters A (January-March), B (April-June), C (July-September) and D (October-December). Maximum number of outbreaks - 106 (34.3%) was recorded in quarter D followed by quarters B - 84 (27.3%), C - 64 (20.7%), and A - 55 (17.7%). Salmonella isolates (253) were recovered from disease outbreaks in broilers from different parts of Haryana. Typical morphology and colony characters on MacConkeys Lactose Agar and Brilliant Green agar, biochemical reactions, serotyping along with antibiogram profiles were able to group these isolates into 3 groups namely Salmonella Gallinarum (183), Salmonella Enteritidis (41) and Salmonella Typhimurium (29). The antibiogram pattern of 183 isolates of S. Gallinarum revealed that most of the isolates were sensitive to gentamicin (76%) followed by amikacin (72%), kanamycin (71%)., Conclusion: FT is prevalent in commercial broiler flocks in different parts of Haryana and is responsible for considerably high morbidity and mortality in affected flocks. Isolation of S. Gallinarum (9, 12:183) from FT cases suggest it to be the primary pathogen, however, isolation of S. Typhimurium and S. Enteritidis from these cases is a major concern. The detection of S. Enteritidis and S. Typhimurium from FT cases assumes significance from public health point of view.

Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum causes fowl typhoid (FT), which results in huge economic losses to poultry farmers in India. We report the draft genome sequence of Salmonella biovar Gallinarum strain VTCCBAA614, isolated from a chicken in an FT affected broiler flock., (Copyright © 2015 Vaid et al.)

Present study was undertaken to investigate the cause of deaths of peafowls in Haryana State. In total, 145 birds were sick and 28 birds were reported dead during July to September 2012. Some of the sick birds were showing signs of shaking of heads, torticollis and paresis. Blood and cloacal swab samples from sick birds along with brain and intestinal tissues from dead birds were collected for further investigation. Although post-mortem examination showed no typical lesions of Newcastle disease virus (NDV) yet raised HI tires against NDV in some serum samples and clinical signs indicated the presence of NDV. One of the brain tissues (NDV/IND2012/01) from the field case was processed and adapted to Vero cell line for virus isolation. The fusion (F) gene based nested RT-PCR (RT-nPCR) confirmed the presence of NDV in all field samples and cell culture isolate. Sequencing of the partial F gene amplicons (216 bp) using the PCR primers as sequencing primers confirmed the PCR results. The deduced amino acid sequences of partial F gene were found to have the amino acid motif (111)GRRQKR/F(117) in the fusion protein cleavage site (FPCS). This amino acid motif is indicative of the velogenic nature of these NDVs. Phylogenetic studies have shown that the virus belonged to class II genotype VII very closely related to virus isolates originated from outbreaks in Western Europe, Israel, Indonesia, Taiwan and India. Phylogenetic grouping of the virus and sequence of FPCS is indicative of pathogenic potential of virus strain circulating in peacocks in Haryana.

Adenoviruses have been isolated from both clinically healthy and diseased birds worldwide. The pathogenic role of most of the FAdVs is still questionable. They can quickly take on the role of opportunistic pathogens when additional factors, particularly concurrent infections, adversely affect the health of the avian host. Immnosuppressing agents especially chicken infectious anemia and infectious bursal disease viruses are known to enhance the pathogenicity of FAdVs upon coinfection. The aim of the present study was to screen for the involvement of FAdV in poultry flocks affected with respiratory disease complex by RT-PCR. The samples were also screened by RT-PCR/PCR for other respiratory pathogens. Thirty two commercial poultry flocks with the history of respiratory disease complex from various parts of India. FAdV nucleic acid could be detected in tissue samples of 13 out of 34 farms investigated. Out of 13 FAdV positive farms, FAdV and CIAV were alone detected in 4/13 (31%) whereas, in other farms more than two respiratory pathogens were detected together. CIAV was detected in all the farms (34/34) investigated. Eosinophilic intranuclear inclusion bodies were noticed in FAdV infected laryngeal and tracheal epithelium under light microscopy. The findings of the study assert that FAdV can play the role of primary respiratory pathogen in immunocompromised birds and also in the presence of other respiratory pathogens.

Augmenter of Liver Regeneration (ALR) is a sulfhydryl oxidase carrying out fundamental functions facilitating protein disulfide bond formation. In mammals, it also functions as a hepatotrophic growth factor that specifically stimulates hepatocyte proliferation and promotes liver regeneration after liver damage or partial hepatectomy. Whether ALR also plays a role during vertebrate hepatogenesis is unknown. In this work, we investigated the function of alr in liver organogenesis in zebrafish model. We showed that alr is expressed in liver throughout hepatogenesis. Knockdown of alr through morpholino antisense oligonucleotide (MO) leads to suppression of liver outgrowth while overexpression of alr promotes liver growth. The small-liver phenotype in alr morphants results from a reduction of hepatocyte proliferation without affecting apoptosis. When expressed in cultured cells, zebrafish Alr exists as dimer and is localized in mitochondria as well as cytosol but not in nucleus or secreted outside of the cell. Similar to mammalian ALR, zebrafish Alr is a flavin-linked sulfhydryl oxidase and mutation of the conserved cysteine in the CxxC motif abolishes its enzymatic activity. Interestingly, overexpression of either wild type Alr or enzyme-inactive Alr(C131S) mutant promoted liver growth and rescued the liver growth defect of alr morphants. Nevertheless, alr(C131S) is less efficacious in both functions. Meantime, high doses of alr MOs lead to widespread developmental defects and early embryonic death in an alr sequence-dependent manner. These results suggest that alr promotes zebrafish liver outgrowth using mechanisms that are dependent as well as independent of its sulfhydryl oxidase activity. This is the first demonstration of a developmental role of alr in vertebrate. It exemplifies that a low-level sulfhydryl oxidase activity of Alr is essential for embryonic development and cellular survival. The dose-dependent and partial suppression of alr expression through MO-mediated knockdown allows the identification of its late developmental role in vertebrate liver organogenesis.

Brucella melitensis Rev 1 organisms were salt-extracted and the cell surface proteins (BCSPs) were found to be mainly 39-42 kDa (group 2 porin proteins) in addition to 31.6, 32.5, 58.5 and 14.7 kDa proteins. DEAE-Sephadex anion-exchange column chromatography of BCSPs yielded fraction 1, which contained one major protein (39.8-42.0 kDa) and a minor protein (31.6 kDa). All these proteins were found to be immunogenic by Western blotting. Fraction 1 along with monophosphoryl lipid A and trehalose dicorynomycolate adjuvants as well as BCSPs alone induced significant (p < or = 0.05) protection in BALB/c mice. Both these immunizing agents produced almost equivalent protection to live B. melitensis Rev 1 vaccine at 15 and 30 days post challenge. Lymphocyte stimulation test as well as delayed-type hypersensitivity reaction revealed that both these preparations induced cell-mediated immune response. These preparations also induced humoral immune response as indicated by indirect ELISA. Neither of the immune responses was significantly less (p < or = 0.05) than that with live B. melitensis Rev 1 vaccine, except that their duration was short.

A study was conducted on 60 broiler chicks of the effect of activated charcoal (200 ppm) on the toxicity of 0.5 ppm aflatoxin B1 (AFB1) in feed fed from day 1 to day 42. Activated charcoal was found to be moderately effective in reducing the harmful effects of AFB1 as assessed by growth response and various biochemical parameters. The feeding of activated charcoal along with AFB1 reduced the inhibitory effect of AFB1 on bodyweights and feed intake. There was also a significant improvement in the serum aspartate aminotransferase, alkaline phosphatase, total proteins, calcium and phosphorus levels. However, no significant improvement was observed in cholesterol levels.

A comparative study of the standard tube agglutination test (SAT), Rose Bengal plate agglutination test and counter immuno-electrophoresis (CIEP) was made on 647 sera from naturally aborting ewes, orchitic, in-contact and apparently healthy sheep with no history of vaccination against brucellosis. No individual test could detect all the 13 known positive reactors (the foetuses of which yielded Brucella melitensis) but by combination of two tests all 13 were positive. The SAT detected more reactors during the early stage of infection while CIEP performed better in later stages of infection. All these tests may be carried out in a field laboratory at very low cost.

An accurate, sensitive, and specific liquid-chromatographic method is described for measuring piperacillin in plasma and urine. Plasma samples deproteinized with two volumes of acetonitrile containing 1.2 mg of the internal standard, p-nitrobenzene sulfonamide, per liter are centrifuged. The clear supernate is evaporated under nitrogen, and the residue is reconstituted in 50 microL of the mobile phase (32/68 by vol acetonitrile/water, adjusted to pH 2.5 with 85% phosphoric acid), of which 10 microL is injected onto a reversed-phase (C-18) column. Urine samples are diluted 10-fold with distilled water, an equal volume of acetonitrile containing 3 mg of the internal standard per liter is added, and 20 microL is chromatographed. Stability studies indicate that storage conditions are critical for both plasma and urine. Piperacillin in plasma is stable at -70 degrees C for at least six weeks, but 100% of it is degraded during the same time at -20 degrees C. Piperacillin in urine is also stable at -70 degrees C for six weeks, but 20% is degraded during six weeks at -20 degrees C.

Present study was undertaken to determine the association of brucellosis with abortions occurring naturally in sheep at an organized local sheep breeding farm. A total of 15 strains of Brucella melitensis biovar I were isolated from the abortion material. Serologically the aborted ewes were positive for brucellosis by one or more tests. During acute infection (abortion), standard tube agglutination test (SAT) detected more positive reactors (70.7%) while counter immunoelectrophoresis (CIE) detected more positive reactors (33.9%) in chronic infection (in-contact and apparently healthy sheep). Personnel handling the abortion material at the farm were found positive clinically as well as serologically for brucellosis. These observations suggest the zoonotic importance of brucellosis.