Molecular detection of respiratory avian mycoplasmosis associated bacterial and viral concurrent infections in the poultry flocks.

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.

Understanding Poor Milling Behavior of Voriconazole from Crystal Structure and Intermolecular Interactions.

The study investigated the milling behavior of voriconazole (VRZ) subjected to particle size reduction using air jet mill at differential air pressures of 5, 6, 7, and 8 bar for five cycles at each pressure. The crystal structure of VRZ was probed for understanding the fracture behavior from crystal packing and intermolecular interactions using molecular modeling tools of attachment energy (Eatt), density functional theory, and energy framework analysis. Upon milling for different cycles, VRZ showed that size reduction from (D90) 20 to 9 µm and 100% particles could not be milled to sizes below 9 µm, with the increase in either the milling intensity or cycle. The milled samples retained the original crystal lattice as evident from consistent melting endotherm (Tm = 130.75 °C); heat of fusion (ΔHf = 96.52 J/g) values; and the plate-shaped morphology. The powder X-ray diffraction pattern of milled samples consistently showed characteristic peaks of stable form B of VRZ. The crystallographic plane (001) was found to be the most prominent slip and the cleavage plane due to least Eatt and weak noncovalent interactions (6.996 kJ/mol) between 3'H and 4'F functional groups of the neighboring planes. The predicted indentation hardness value of 228.67 MPa further indicated toward the plastic nature of VRZ crystals. Corroborating outcomes from the different molecular modeling tools for VRZ, cleavage along the plane (001) was determined to be energetically favorable, whereas cleavage of isotropic 2D molecular sheets was energetically unfavorable. As milling proceeds and crystal reduces in size, contact surface area and overall interaction energy decrease contributing to plastic behavior of the crystal. It was concluded that crystal plasticity and isotropic 2D molecular sheets along with the orientation of particles to the direction of stress and attrition energy during air jet milling are contributing factors for nonuniform size reduction of VRZ particles.

Insights on Mycoplasma gallisepticum and Mycoplasma synoviae infection in poultry: a systematic review.

Avian mycoplasmosis mainly caused by Mycoplasma gallisepticum and M. synoviae is an economically important disease of poultry industry. It causes huge economic losses in terms of decrease in weight gain, feed conversion efficiency, egg production, hatchability; increase in embryo mortality, carcass condemnation, prophylaxis and treatment cost in broiler, layer and breeder flocks. The disease is caused by four major pathogenic mycoplasmas viz., M. gallisepticum (MG), M. synoviae (MS), M. meleagradis (MM) and M. iowae (MI). The MG and MS are World Organization for Animal Health listed respiratory pathogens. MG causes chronic respiratory disease in chicken and infectious sinusitis in turkey; however, MS causes synovitis and airsacculitis in birds. The infection is transmitted both horizontally and vertically. Prevention and control measures of avian mycoplasmosis mainly comprises of biosecurity, treatment and vaccination. For vaccination of birds, inactivated bacterins, live attenuated and/or recombinant live poxvirus vaccines are commercially available against MG and MS infection. The present systematic review summarizes the different epidemiological studies carried out on MG and MS infection in poultry in different geographical locations of India and abroad over the last decade (2010-2020), economic impact, diagnosis and prevention and control.

Coxiella burnetii in cattle and their human contacts in a gaushala (cattle shelter) from India and its partial com1 gene sequence-based phylogenetic analysis.

Q fever caused by Coxiella burnetii is an important zoonosis and has great public health significance. A total of 905 clinical samples from 387 cattle [serum (n = 387); vaginal swabs (n = 387); milk (n = 131)] and 59 serum samples from humans were collected from gaushala (cattle shelter) and screened for anti-C. burnetii IgG antibodies in the sera using an indirect-ELISA kit. Further, the samples were tested for C. burnetii DNA employing TaqMan real-time and conventional PCR assays targeting the com1 gene. In ELISA, 9.56% and 6.78% of animal and human sera samples were positive for anti-C. burnetii antibodies, respectively. Upon pathogen detection, 3.87% sera, 1.81% vaginal swabs, and 6.87% milk samples from cattle tested positive in TaqMan real-time PCR and 1.55% sera, 0.52% vaginal swabs, and 3.05% milk samples were found positive in conventional PCR. In humans, one serum sample was positive in both the PCR assays. The PCR positive samples (n = 12) were partially sequenced and the phylogenetic tree was constructed using com1 gene sequences (n = 42) from a different host and geographical areas. The study highlights infection of cattle and their human contacts in gaushala and identifies relationships between strains identified in the gaushala and those in other parts of the globe.

Prevalence, molecular characterization, and antimicrobial resistance profile of Clostridium perfringens from India: A scoping review.

Clostridium perfringens is one of the most important foodborne pathogens that causes histotoxic diseases and intestinal infections in both humans and animals. The present scoping review has been designed to analyze the literature published during 2000-2021 from India on the prevalence, molecular characterization, and antimicrobial resistance profile of C. perfringens isolates recovered from humans, animals, animal-based foods, and associated environmental samples. The peer-reviewed articles retrieved from four electronic databases (Google Scholar, PubMed, Science Direct, and Web of Science) were assessed using PRISMA-ScR guidelines. A total of 32 studies from India were selected on the basis of their relevance and inclusion criteria. The overall prevalence of C. perfringens among domestic animals having history of clinical symptoms and among healthy animals was found to be 65.8% (508/772) and 42.8% (493/1152), respectively. The pathogen was also detected in clinically affected wild animals (75%), healthy wild animals (35.4%), and captive birds (24.5%). The detection of C. perfringens among poultry having necrotic enteritis and among healthy birds was found to be 66.8% (321/480) and 25.6% (80/312), respectively. The detection of pathogen among animal-based foods (i.e., meat, milk, and fish and their products) and environmental samples depicted a prevalence of 20.8% (325/1562) and 30.2% (23/76), respectively. However, the prevalence of C. perfringens among humans having history of diarrhea and among healthy humans was found to be 25% (70/280) and 23.2% (36/155), respectively. The genotyping of C. perfringens isolates revealed that toxin type A was found to be the most prevalent genotype. Along with the alpha toxin gene (cpa), beta (cpb), epsilon (etx), iota (itx), enterotoxin (cpe), beta-2 toxin (cpb2), and NetB (netB) toxins were also detected in different combinations. Antimicrobial resistance profile of C. perfringens isolates recovered from different sources demonstrated that the highest resistance was detected against sulphonamides (76.8%) and tetracycline (41.3%) by phenotypic and genotypic detection methods, respectively. Comprehensive scientific studies covering different geographical areas at the human-animal-environment interface are crucial to generalize the real magnitude of C. perfringens-associated problem in India and for establishing a reliable database.

Antimicrobial efficacy of Cecropin A (1-7)- Melittin and Lactoferricin (17-30) against multi-drug resistant Salmonella Enteritidis.

The present study evaluated intracellular antibacterial efficacy of two short-chain cationic antimicrobial peptides (AMPs) namely, Cecropin A (1-7)-Melittin and lactoferricin (17-30) against three field strains of multi-drug resistant Salmonella Enteritidis. Initially, antimicrobial ability of both the AMPs was evaluated for their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against multi-drug resistant S. Enteritidis strains. Besides, the AMPs were evaluated for its in vitro stability (high-end temperatures, proteases, physiological concentrations of cationic salts and pH) and safety (haemolytic assay in sheep erythrocytes; cytotoxicity assay in murine macrophage RAW 264.7 cell line and human epithelioma HEp-2 cell line and beneficial gut lactobacilli). Later, a time-kill assay was performed to assess the intracellular antibacterial activity of Cecropin A (1-7)-Melittin and lactoferricin (17-30) against multi-drug resistant S. Enteritidis in RAW 264.7 and HEp-2 cells. The observed MBC values of Cecropin A (1-7)-Melittin and lactoferricin (17-30) against multi-drug resistant S. Enteritidis (128 µM; 256 µM) were generally twice or four-fold greater than the MIC values (64 µM). Further, both the AMPs were found variably stable after exposure at high-end temperatures (70 °C and 90 °C), protease treatment (trypsin, proteinase K, lysozyme), higher concentration of physiological salts (150 mM NaCl and 2 mM MgCl2) and hydrogen ion concentrations (pH 4.0 to 8.0). Both the AMPs were found non-haemolytic on sheep erythrocytes, revealed minimal cytotoxicity in RAW 264.7 and HEp-2 cells, and was tested safe against beneficial gut lactobacilli (L. acidophilus and L. rhamnosus). Intracellular bacteriostatic effect with both cationic AMPs against multi-drug resistant S. Enteritidis was evident in RAW 264.7 cells; however, in both the cell lines, the significant bactericidal effect was not observed (P > 0.05) with both cationic AMPs understudy against multi-drug resistant S. Enteritidis. Based on the results of the present study, both the cationic AMPs understudy may not be useful for the intracellular elimination of multi-drug resistant S. Enteritidis; hence, further studies such as conjugation of these AMPs with either cell-penetrating peptides (CPP) and/or nanoparticles (NPs) are warranted.

Molecular Understanding and Implication of Structural Integrity in the Deformation Behavior of Binary Drug-Drug Eutectic Systems.

In eutectic, a lamellar microstructure offers better tableting than that of the nonreacted physical mixture. However, bulk deformation remains elusive in two binary eutectics. We hypothesized that the binary eutectic of a drug with different components, having different H-bonding dimensionalities and crystal structure, shall allow the understanding of the structural integrity in the bulk deformation behavior. The shearing molecular solid (FXT Q) shared a common composition with the viscoelastic crystal (ASP I) and brittle (PCM I), forming EM-1 (ϕ1 = 41.27:58.73% w/w) and EM-2 (ϕ2 = 41.10:58.90% w/w), respectively. The excess thermodynamic functions were contributed by high energy microstructures (nonbonding interactions) along incoherent phase boundaries (visualized under CLSM). The energy dispersive analysis enabled the recognition of the relative distribution of higher atoms over the heterogeneous surface. EM-1 (FXT Q-ASP I) demonstrated higher compressibility, tensile strength, and compactibility (CTC profile) compared to those of EM-2 (FXT Q-PCM I) over a range of applied compaction pressures. The lower true yield strength (σ0(EM-1) = 138.66 MPa) of EM-1 as compared to that of EM-2 (σ0(EM-2) = 166.66 MPa) suggested a better deformation performance and incipient plasticity quantified from the "out-of-die" Heckel analysis. From Ryshkewitch analysis, the tensile strength at zero porosity (τ01 = 3.83 MPa) was predicted to be higher for EM-1 than EM-2 (τ02 = 2.54 MPa). The higher bonding strength of EM-1 was contributed to the additional influence of true density and isotropic van der Waals interactions of ASP I (0D). In contrast, EM-2 demonstrated lower compressibility and compactibility, having herringbone molecular packing of PCM I (1D) with a common shearing component (FXT Q (1D)). This study confirmed that the intrinsic deformational and chemical nature of the second component defined the compressibility and compactibility tendency to a greater extent in the tableting performance of conglomerates of crystalline solid solution.

Revealing the Role of Structural Features in Bulk Mechanical Performance of Ternary Molecular Solids of Isoniazid.

Mechanical performance in ternary (3n) molecular solids has been rarely studied, and hence it is an interesting topic of investigation in the direct compression method of tableting. The structural features of 3n-eutectic (3n-Eu: INZ-ADP-NIC) and 3n-cocrystal (3n-Co: INZ:SUC:NIC) were explored to understand the bonding area-bonding strength (BA-BS) interplay. Higher compressibility and lower values of the Heckel parameter of 3n-Co as compared to 3n-Eu suggested its better deformation behavior, with BA being the predominant factor. The higher tensile strength and Walker analysis indicated a higher compressibility coefficient ( W) and lower pressing modulus ( L) for 3n-Eu, which was consistent with its better tabletability over 3n-Co. The higher compressibility and plastic energy, and higher value of L of 3n-Co, were attributed to the facile propagation (⟨-1' 0' 5'⟩) of the shearing molecular slip (-1 0 5) when subjected to the external mechanical stress. Thus, the overall higher tableting performance of 3n-Eu over 3n-Co was found due to the predominant BS and limited contribution of BA. The latter was the dominant factor in 3n-Co. Cohesive interactions, like the 3D mechanically interlocked structure of conglomerates of 3n-Eu, contributed toward the higher BS. Moreover, the prediction of better tabletability solely based on crystallographic feature slip planes (0D/1D/2D H-bonded layer (h k l) ⊥ vdW interactions) is warranted in pharmaceutical molecular solids. Eutectics with varying microstructural variants ( nLα + nLß + nLγ) may open up the opportunity to manipulate the physicomechanical performance.

Molecular characterization and antimicrobial resistance profile of Clostridium perfringens type A isolates from humans, animals, fish and their environment.

The study was aimed to characterize, and determine antibiogram of C. perfringens type A isolated from the feces of human and animal diarrhoeal cases, as well as healthy animals, meat of pigs and goats, gills and intestine of fish and samples from fish pond. A total of 460 samples, including human diarrhoeal cases (n = 130); diarrhoeal cases of pig (n = 52) and goat (n = 50); fecal samples from healthy pig (n = 50) and goat (n = 50); meat samples viz. pork meat (n = 52); goat meat (n = 50) and fish including their environmental sources (n = 26) were used for isolation and identification of C. perfringens type A. All the biochemically confirmed isolates were positive for species-specific 16S rRNA and cpa genes by PCR assays. Toxinotyping of C. perfringens type A isolates showed that overall prevalence of C. perfringens type A with only cpa+ gene was 43.2%; with cpa+ and cpb2+ genes was 45.4%; with cpa+ and cpe+ genes was 4.9%; however, with cpa+, cpb2+and cpe+ genes was 6.6%. Antimicrobial susceptibility testing revealed that 83.7% of isolates were resistant to three or more antibiotics.

Molecular and serological screening of coxiellosis in bovines with reproductive disorders in dairy farms.

Coxiella burnetii infection is an emerging/re-emerging public health problem affecting several countries worldwide. In India, the disease is mainly underdiagnosed, creating hindrances in its effective control. This study investigated the occurrence of C. burnetii among apparently healthy cattle and cattle with a history of reproductive disorders by both PCR and indirect-ELISA. A total of 731 clinical samples (serum: 531, and vaginal swabs as well as blood: 100 each) from 531 cattle were screened for coxiellosis. The serum, blood, and vaginal swabs each collected from 100 cattle with a history of reproductive disorders were screened using Com1-PCR, Trans-PCR, and indirect-ELISA. Conversely, serum samples obtained from apparently healthy cattle were exclusively screened using indirect ELISA. None of the samples tested could detect C. burnetii in PCR assays, while 13.37 % of serum samples were found to be seropositive in i-ELISA. Seropositivity noted among clinically healthy and those suffering from reproductive disorders were 12.76 % and 16 %, respectively, exhibiting a non-significant difference observed between these two categories. The obtained results suggested that the occurrence of coxiellosis did not differ significantly between clinically healthy animals and those with reproductive disorders; hence, in farms affected with C. burnetii infection, screening healthy and symptomatic animals is crucial to implement appropriate preventive measures.

Surveillance and response strategies for zoonotic diseases: a comprehensive review.

Out of all emerging infectious diseases, approximately 75% are of zoonotic origin, with their source often traced back to animals. The emergence of zoonoses is driven by a complex interplay between anthropogenic, genetic, ecological, socioeconomic, and climatic factors. This intricate web of influences poses significant challenges for the prediction and prevention of zoonotic outbreaks. Effective coordination and collaboration among the animal, human, and environmental health sectors are essential for proactively addressing major zoonotic diseases. Despite advancements in surveillance and diagnostic practices, the emergence of zoonoses continues to be a pressing global concern. Therefore, prioritizing zoonotic disease surveillance is of paramount importance as part of a comprehensive disease prevention and containment strategy. Furthermore, evaluating existing surveillance systems provides insights into the challenges faced, which can be mitigated through implementation of One Health principles involving relevant stakeholders. To initiate multisectoral partnerships, it is crucial to identify the priorities and core themes of surveillance systems with equitable inputs from various sectors. Strengthening surveillance, promoting data sharing, enhancing laboratory testing capabilities, and fostering joint outbreak responses in both the human and animal health sectors will establish the necessary infrastructure to effectively prevent, predict, detect, and respond to emerging health threats, thereby reinforcing global health security. This review assesses existing surveillance approaches by offering an overview of global agencies engaged in monitoring zoonoses and outlines the essential components required at the human-animal-environment interface for designing comprehensive surveillance networks. Additionally, it discusses the key steps necessary for executing effective zoonotic disease surveillance through a One Health approach, while highlighting the key challenges encountered in establishing such a robust surveillance system.

Rapid and specific detection of Mycoplasma gallisepticum and Mycoplasma synoviae infection in poultry using single and duplex PCR assays.

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.

Comparative evaluation of indirect-ELISA and DOT blot assay for serodetection of Mycoplasma gallisepticum and Mycoplasma synoviae antibodies in poultry.

Avian mycoplasmosis, mainly caused by Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) is an economically important disease of the poultry industry. The present study was aimed to develop whole cell based indirect-ELISA (i-ELISA) and DOT blot assay (DOT-ELISA) as rapid, sensitive, specific and economical sero-detection tests for MG and MS. A total of 306 blood samples were collected from birds slaughtered at local meat shops of different districts of Haryana, India to detect MG and MS antibodies. Sonicated antigens prepared from freshly grown culture of MG and MS were used to develop i-ELISA and DOT blot assay. In i-ELISA, 50.32% and 61.76% serum samples were found to be positive for MG and MS antibodies, respectively. However in DOT blot assay, 41.83% and 53.92% serum samples were found positive for MG and MS antibodies, respectively. The relative diagnostic sensitivity and specificity of DOT-ELISA were measured considering i-ELISA as a reference test. The relative diagnostic sensitivity of the DOT blot assay was found to be 69.48% and 82.01%; whereas relative diagnostic specificity was 86.18% and 91.45% for the detection of MG and MS antibodies, respectively. The developed serological assays may be used as rapid and economical diagnostic tools for large scale screening of poultry sera for MG and MS antibodies.

Current perspectives on the occurrence of Q fever: highlighting the need for systematic surveillance for a neglected zoonotic disease in Indian subcontinent.

Coxiellosis or Q fever is an important global occupational zoonotic disease caused by one of the most contagious bacterial pathogens - Coxiella burnetii, which ranks one among the 13 global priority zoonoses. The detection of C. burnetii infection is exhibiting an increasing trend in high-risk personnel around the globe. It has increasingly been detected from foods of animal origin (including bulk milk, eggs, and meat) as well as tick vectors in many parts of the world. Coxiellosis is reported to be an important public health threat causing spontaneous abortions in humans and potential reproductive failure, which would result in production losses among livestock. Further, comprehensive coverage of the reports and trends of Q fever in developing countries, where this infection is supposed to be widely prevalent appears scarce. Also, the pathogen remains grossly neglected and underreported. Moreover, policymakers and funding agencies do not view it as a priority problem, especially in the Indian subcontinent, including Sri Lanka, Bhutan, Pakistan, Nepal, Bangladesh and Maldives. Here, we review the occurrence and epidemiology of the disease in a global context with special emphasis on its status in the Indian subcontinent.

Molecular Investigation of the Status of Ticks on Infected Cattle for Coxiella burnetii in India.

PURPOSE: In Indian subcontinent, the epidemiological studies on the status of ticks in the transmission of Coxiella burnetii have not been explored comprehensively. The objective of the present study was to investigate the status of ticks for C. burnetii among coxiellosis positive cattle. METHODS: The present study was carried out in three locations of the northern states of India. A total of 1648 tick samples were collected from the tick infested cattle (n = 146) that were tested positive for coxiellosis by indirect serum-ELISA assay and/or the trans-PCR assay. The tick samples were screened using the trans-PCR assay targeting species-specific IS1111 transposase gene of C. burnetii. The sequencing of PCR products was planned to differentiate C. burnetii and Coxiella-like bacteria (CLB). RESULTS: The collected ticks were identified as Rhipicephalus microplus (n = 1049), Hyalomma anatolicum (n = 416), and Hyalomma spp. (n = 183). On molecular investigation, none of the collected tick samples were found to be positive for the IS1111 gene. CONCLUSION: The findings of the present study ruled out the involvement of ticks in circulation of the pathogen within the cattle population that were screened. However, extensive epidemiological studies are needed to conclusively rule out or establish the role of ticks as a competent vector for C. burnetii transmission in cattle and other hosts.

Apparent prevalence and risk factors of coxiellosis (Q fever) among dairy herds in India.

Coxiella burnetii is a highly infectious zoonotic pathogen infecting wide range of mammals, including humans. In the present study, a total of 711 blood samples from bovines [cattle (n = 543) and buffaloes (n = 168)] from eight farms at different geographical locations in India were screened for C. burnetii targeting the IS1111 and the com1 genes. The anti-C. burnetii antibodies in serum samples were detected using indirect-ELISA kits. Also, a total of 21 parameters pertaining to animal health and farm management were identified to assess their role as possible risk factors for coxiellosis among the targeted farms. The apparent prevalence (positive for PCR and/or ELISA) for coxiellosis was reported to be 24.5% in cattle and 8.9% in buffaloes. In cattle, the detection rate of C. burnetii employing the IS1111 gene (8.5%) was found to be significantly higher (p<0.05) as compared to the com1 (6.5%) gene. The seropositivity by ELISA was higher among cattle (17.7%) than in buffaloes (8.3%). Further, on univariable analysis of risk factors, species (cattle) (OR:3.31; 95%CI:1.88-5.82), inadequate floor spacing (OR:1.64; 95%CI:1.10-2.43), mastitis (OR:2.35, 95%CI:1.45-3.81) and reproductive disorders (OR:2.54; 95%CI:1.67-3.85) were significantly (p<0.05) having high odds for coxiellosis. The multivariable logistic regression analysis of the animal level risk factors revealed that species and age were found to be significantly associated with coxiellosis. However, since the number of screened farms is limited; further research is needed with a higher number of animals to confirm the farm level odds ratio of risk factors. Quarantine and biosecurity measures including farm hygiene operations were observed to be inadequate and also the lack of awareness about coxiellosis among the farm workers. In absence of vaccination program for coxiellosis in India, robust surveillance, farm biosecurity measures and the awareness for the disease among risk groups can play an important role in the disease prevention and subsequent transmission of the pathogen.

Comparison of two new in-house Latex Agglutination Tests (LATs), based on the DnaK and Com1 synthetic peptides of Coxiella burnetii, with a commercial indirect-ELISA, for sero-screening of coxiellosis in bovines.

The in-house developed DnaK and Com1 synthetic peptide-based Latex Agglutination Tests (LATs) were comparatively evaluated with commercial indirect-ELISA kit, to provide a rapid, economical and onsite field applicable test for seroscreening of coxiellosis in bovines.

Development of the Com1 synthetic peptide-based Latex Agglutination Test (LAT) and its comparative evaluation with commercial indirect-ELISA for sero-screening of coxiellosis in cattle.

A novel Com1 synthetic peptide-based latex agglutination test (LAT) was developed and evaluated against commercial ELISA kit for sero-screening of coxiellosis in cattle. The developed test is economical, has field applicability and can serve as an important rapid tool for sero-screening of coxiellosis in cattle.

Loop-mediated isothermal amplification assay for detection of Coxiella burnetii targeting the com1 gene.

We developed the com1 gene based loop-mediated isothermal amplification (LAMP) assay for the detection of Coxiella burnetii and validated it by screening DNA isolated from serum samples collected from animals and humans. The detection of Coxiella by LAMP assay was comparable with the com1 based-PCR.

Pathology and polymerase chain reaction detection of ovine progressive pneumonia (maedi) cases in slaughtered sheep in India.

AIM: The small ruminant lentiviruses are known to cause maedi-visna (MV) and caprine arthritis - encephalitis in sheep and goats, typically affecting joints, udder, lungs, and the central nervous system. The diagnosis usually involves serology, clinical signs, immunohistochemistry, and polymerase chain reaction (PCR). In the present study, the histopathologically positive pneumonia cases of MV were confirmed by PCR in lung tissue probably for the first time in India. MATERIALS AND METHODS: A total of 888 lungs of adult sheep, aged between 2 and 5 years, were screened during slaughter, of which 121 were found to have pneumonic lesions. The tissues from each pneumonic lung including associated lymph nodes were collected in 10% neutral buffered formalin for histopathology. The frozen tissues of the same were also collected and stored at -20°C for PCR confirmation. RESULTS: Three of 121 cases of pneumonic lungs of sheep revealed gross and histopathological lesions suggestive of maedi or ovine progressive pneumonia infection. These 3 cases were further confirmed by PCR technique that amplified 291-base pair DNA in the long terminal repeat sequence of MV provirus. CONCLUSION: This study suggests the low occurrence of MV virus (MVV) infection in India in naturally affected sheep based on pathomorphological lesions and using the molecular tool of PCR detection of the virus in tissues. Further, a combination of pathomorphology or/and PCR testing might be optimal for detecting the animals infected with MVV.