Unsupervised machine learning, QSAR modelling and web tool development for streamlining the lead identification process of antimalarial flavonoids.

Identification of lead compounds with the traditional laboratory approach is expensive and time-consuming. Nowadays, in silico techniques have emerged as a promising approach for lead identification. In this study, we aim to develop robust and predictive 2D-QSAR models to identify lead flavonoids by predicting the IC50 against Plasmodium falciparum. We applied machine learning algorithms (Principal component analysis followed by K-means clustering) and Pearson correlation analysis to select 9 molecular descriptors (MDs) for model building. We selected and validated the three best QSAR models after execution of multiple linear regression (MLR) 100 times with different combinations of MDs. The developed models have fulfilled the five principles for QSAR models as specified by the Organization for Economic Co-operation and Development. The outcome of the study is a reliable and sustainable in silico method of IC50 (Mean ± SD) prediction that will positively impact the antimalarial drug development process by reducing the money and time required to identify potential antimalarial lead compounds from the class of flavonoids. We also developed a web tool (JazQSAR, https://etflin.com/news/4) to offer an easily accessible platform for the developed QSAR models.

Development of a novel one-step triplex PCR assay for the simultaneous detection of porcine circovirus type 2, porcine parvovirus and classical swine fever virus in a single tube.

Co-infection of multiple pathogens complicates diagnosis, treatment and preventive measures based on clinical signs. Therefore, reliable diagnostic tool for timely reporting of suspected diseases is very much essential. A novel one-step triplex PCR assay was developed and evaluated for simultaneous detection of three important viruses namely porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classical swine fever virus (CSFV) involved in reproductive problems in pigs. Each of the three pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses. The multiplex PCR assay was found to be sensitive in detecting at least 300 pg of viral genomic DNA or RNA from a mixture of three viruses in a reaction. No amplification was obtained from other common viruses or pathogens, such as porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), porcine group A rotavirus (PoRVA), Escherichia coli and Staphylococcus aureus thereby indicating that the developed multiplex PCR has high specificity. Because of the sensitivity and specificity, the developed multiplex PCR assay will be a useful tool for clinical diagnosis of mixed infections of DNA and RNA viruses in pigs.

Development of a rapid loop-mediated isothermal amplification (LAMP) assay for visual detection of porcine parvovirus (PPV) and its application.

Porcine parvovirus (PPV) infection is one of the most important causes of reproductive failure in pigs impacting the piggery industry globally with huge economic losses. A cost-effective, simple, rapid, specific, and sensitive method is critical for monitoring PPV infection on pig farms. The main aim of the present study was to develop and evaluate a loop-mediated isothermal amplification (LAMP) assay for rapid visual detection of porcine parvovirus (PPV) in pigs. A set of six LAMP primers including two outer primers, two inner primers, and two loop primers were designed utilizing the conserved region of capsid protein VP2 gene sequences of PPV and was applied for detection of PPV from porcine samples. Time and temperature conditions for amplification of PPV genes were optimized to be 30 min at 63 °C. The developed assay was ten-fold more sensitive than conventional PCR with analytical sensitivity of 20 pg and 200 pg, respectively. This is the first report of detection of PPV by LAMP assay from India. The assay did not cross-react with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), or classical swine fever virus (CSFV). The LAMP assay was assembled into a LAMP assay kit of 20 reactions and was validated in different laboratories in India. The newly developed LAMP assay was proved to be a specific, sensitive, rapid, and simple method for visual detection of PPV which does not require even costly equipments for performing the test. It complements and extends previous methods for PPV detection and provides an alternative approach for detection of PPV.

Virulence associated gene profiling and antimicrobial resistance pattern of Streptococcus suis isolated from clinically healthy pigs from North East India.

This study revealed the prevalence of Streptococcus suis in 20·39% clinically healthy pigs from North East India. All these isolates were screened for the presence of virulence- associated genes such as suilysin (sly), muramidase released protein (mrp), extracellular protein factor (epf) and arginine deiminase (arcA). Of these 62 isolates, 29 isolates carried mrp gene, 17 isolates carried sly gene, 57 isolates carried arcA gene, whereas all isolates were negative for epf gene. The most prevalent genotype was mrp- sly- epf- arcA+ (45·16%) followed by genotypes mrp+ sly- epf- arcA+ (27·41%), mrp+ sly+ epf- arcA+ (19·35%) and mrp- sly+ epf- arcA- (8·06%). High frequency of resistance was observed for antimicrobials such as tetracycline (93·54%), clindamycin (91·93%), co-trimoxazole (88·70%) and erythromycin (85·48%). Antimicrobial resistance patterns of the S. suis isolates revealed 16 resistance groups (R1 to R16), where 93·54% isolates showed multi-drug resistance (≥3 antimicrobial agents). It has also been observed that 57 (91·93%) isolates were resistant to at least four antimicrobials. The most predominant resistance pattern observed was CD-COT-E-TE, which accounted for 38·70% of the isolates. The occurrence of relatively high levels of resistance of S. suis to some antimicrobials (e.g., macrolides, tetracyclines, and sulphonamides) as observed in this study may represent a human health concern. In addition, a relatively higher percentage of S. suis isolated from clinically healthy pigs indicates a carrier status with risk of dissemination to other pigs in the herd as well as to humans.

Development and application of a triplex-PCR assay for rapid detection of methicillin-resistant Staphylococcus aureus from pigs.

A triplex-PCR assay was developed and evaluated for rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102  CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex-PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false-positive amplification was observed, indicating the high specificity of the developed triplex-PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin-sensitive S. aureus, coagulase-negative methicillin-resistant staphylococci and coagulase-negative methicillin-sensitive staphylococci.

Haemato-biochemical alterations and oxidative stress associated with naturally occurring porcine circovirus2 infection in pigs.

Porcine circovirus2 (PCV2) infection in pigs is one of the major causes of economic loss to the farmers in terms of low production, slow growth and increase post-weaning mortality rate. The effect of PCV2 infection on haemogram, serum biochemical profile and oxidant/anti-oxidant status is not well established in pigs. In the present study, haemogram, serum biochemical profile and oxidant/anti-oxidant status were assessed in pigs confirmed positive for PCV2 infections as evidenced by commercially available enzyme-linked immunosorbent assay kit (n = 151) and polymerase chain reaction (PCR) (n = 42) among a total of 306 number of pigs included in the study. Non-infected healthy pigs (n = 6) served as healthy control. The total erythrocyte count (TEC), haemoglobin (Hb), packed cell volume (PCV), total leukocyte count (TLC), differential leukocyte count (DLC) and thrombocyte count were measured. The levels of total protein, albumin, globulin, total bilirubin, direct bilirubin, blood urea nitrogen (BUN), creatinine and glucose and enzymes viz. alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP) were measured. Oxidative stress indicators such as plasma malondialdehyde (MDA) and total anti-oxidant activity (TAOA) were measured using commercially available kits. The mean values of TLC, lymphocytes and thrombocyte count were significantly (P < 0.05) low in PCV2-infected pigs. The levels of globulin, AST, GGT, BUN and creatinine were significantly increased (P < 0.05) whereas levels of albumin and glucose significantly (P < 0.05) decreased in PCV2-infected pigs. The significant increase (P < 0.05) in MDA level and significant decrease (P < 0.05) in TAOA level were noticed in PCV2-infected animals as compared with healthy control. The present study supports immunosuppression, possible multiple organ damage and oxidative stress associated with naturally occurring PCV2 infection in pigs. Timely vaccination and managemental practices can reduce PCV2 infection in farms. In spite of many research studies, there is still paucity of detailed systemic study on haemato-biochemical alteration and oxidative stress associated with PCV2 infection.

The effect of temperature, composition and alcohols on the microstructures of catanionic mixtures of sodium dodecylsulfate and cetyltrimethylammonium bromide in water.

The influence of mixing protocol, composition, temperature, ageing and added alcohols on the characteristics of the microstructures of sodium dodecylsulfate (SDS) + cetyltrimethylammonium bromide (CTAB) mixtures has been investigated in this paper. In this catanionic mixture (1 weight% total surfactant content) temperature induced microstructural transition occurs, which is (i) a micelle-to-vesicle transition (MVT) if αSDS (mole fraction of SDS) = 0.7, 0.8 or 0.9 and (ii) a vesicle-to-micelle transition (VMT) if αSDS = 0.1, 0.2 or 0.3. In the mixture of αSDS = 0.7, specific conductivity and dynamic light scattering measurements also support the occurrence of MVT. Transition electron microscopy and small angle neutron scattering measurements were also made to assess the characteristics of the microstructures. Alcohols added to the mixture of αSDS = 0.7 reduced the size of the vesicle, while only monohydric alcohols suppressed the temperature induced transition indicating that the number and location of -OH groups of the alcohols have a dramatic modulating influence on the structural transition occurring in catanionic mixtures. The influence of the alcohols is explained in terms of changes produced in the dielectric constant and hydrophobicity of the medium.

Molecular characteristics of Clostridium difficile isolates from human and animals in the North Eastern region of India.

A total of 1034 samples were collected from different sources and C. difficile was isolated from 18 (9.04%) of 199 human, 9 (4.89%) of 184 cattle, 29 (12.44%) of 233 pig, and from 23 (13.94%) of 165 poultry samples. Variations were observed on the rate of isolation according to age and clinical conditions (diarrhoea). None of the samples from cow, sheep, goat, local chicken, and wild animals yielded any C. difficile. Out of those isolates, 8, 2, 19 and 6 isolates from human, cattle, pig and poultry, respectively were toxigenic. The toxigenic isolates carried both tcdA, and tcdB (A+B+) and most of the human and the pig isolates were also positive for binary toxin genes (cdtA and cdtB). The A+B+ isolates belonged to three different toxinotypes (0, VI and XXXIII). Human and pig A+B+ isolates belonged to three (045, 126 and ACD 019) and four (046, 087, 126 and ACD 011) different ribotypes, respectively and the ribotypes of two cattle isolates were 014 and ACD 010. Six A+B+ avian isolates belonged to six different ribotypes (014, 087, SLO 134, SLO 160, ACD 012, ACD 014). The non-toxigenic isolates from human, cattle, pig and poultry were grouped into 7, 4, 4 and 7 different ribotypes, respectively. PFGE analysis could not differentiate similar ribotypes/toxinotypes of toxigenic isolates. All the toxigenic isolates showed cytopathic effect on Vero and Hela cell monolayers at 1:100 dilutions of cell-free culture supernatants within 18-20 h of inoculation.

SCC mec typing and antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) from pigs of Northeast India.

Staphylococcus aureus is one of the most important pathogens of both humans and animal. Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that causes serious infections both in hospitals and communities due to its multidrug resistance tendency. This study was undertaken to characterize the MRSA isolates from pigs and to determine the antimicrobial resistance of these isolates. Forty nine MRSA strains (one strain per positive pig) isolated from pigs of Northeast India were characterized by SCCmec typing and antimicrobial resistance. The overall prevalence of MRSA was 7.02 % with the highest prevalence recorded in pigs aged 1-3 months (P = 0.001) and in nasal samples (P = 0.005). Two SCC mec types (type III and V) were found in Indian pigs with predominance of type V. All isolates were resistant to penicillin. Seventeen resistance groups were observed where 87.75 % isolates showed multidrug resistance (showed resistance to three or more classes of antimicrobials). The most predominant resistance pattern observed was Oxytetracycline + Penicillin + Sulfadiazine + Tetracycline accounting 12.24 % of the isolates. The present study contributes to the understanding of characteristics and antimicrobial resistance of porcine MRSA isolates which in turn will help in devising strategy for the control of this pathogen. Findings of the study also throw light on multidrug resistance MRSA and emphasize the need for judicious use of antimicrobials in animal practice.

Isolation and characterization of Clostridium difficile from pet dogs in Assam, India.

One hundred and seventeen faecal samples from pet dogs (pup = 21 and adult = 96) brought for treatment to a veterinary clinic were examined for Clostridium difficile. A total of 16 (13.67%) samples were positive. Nine (56.25%) isolates were obtained from 17 adult dogs undergoing antibiotic treatment and this was significantly higher (p < 0.01) as compared to isolates from dogs without antibiotic treatment. Ten isolates (62.5%) were toxigenic (all toxinotype 0) and six were non-toxigenic. None of the isolates were positive for binary toxin genes. PCR ribotyping revealed three different ribotypes (012, 014 and 046) among A(+)B(+) isolates and five different ribotypes (010, SLO 131, and ACD 001 to ACD 003) among A(-)B(-) isolates. The PFGE analysis of toxigenic isolates revealed three different pulsotypes corresponding to the PCR ribotypes.

Development of a novel multiplex PCR assay for rapid detection of virulence associated genes of Pasteurella multocida from pigs.

UNLABELLED: As the pathogenicity of Pasteurella multocida is associated with various virulence factors (VFs), the aim of the study was to develop a novel multiplex PCR (m-PCR) assay for the rapid detection of important virulence associated genes (VAGs) of P. multocida isolates from pigs. The target recognized VFs used in the study were diverse adhesins (ptfA and pfhA), toxins (toxA), siderophores (tonB and hgbA), sialidases (nanB, nanH) and outer membrane proteins (ompA, ompH, oma87 and plpB). The primers for the genes encoding these VFs were designed by primer3 software (http://bioinfo.ut.ee/primer3-0.4.0/) using gene sequences available in Genbank. The detection limit of the developed assay was 10(2)  CFU ml(-1) . The m-PCR did not produce any nonspecific amplification products when tested against Bordetella bronchiseptica which also commonly infects pigs. We applied m-PCR to the field samples, and the results obtained were the same as the single PCR results. The developed assay would be very useful for veterinary diagnostic laboratories and for others interested in the rapid virulence profiling of porcine P. multocida isolates circulating in the piggeries. SIGNIFICANCE AND IMPACT OF THE STUDY: The study reports the development and evaluation of a novel multiplex PCR assay for the rapid detection of 11 important VAGs of Pasteurella multocida isolates from pigs. Rapid and simultaneous detection of recognized VFs of the organism are essential to know the virulo-types of P. multocida isolates circulating in the piggeries. The developed novel assay will be very useful for the rapid detection of VAGs of P. multocida isolates from pigs.

Predominance of genotype 1.1 and emergence of genotype 2.2 classical swine fever viruses in north-eastern region of India.

Classical swine fever (CSF) is a highly contagious and the most important disease of pigs worldwide.CSF is enzootic in pig herds in India and continues to cause huge economic losses to pig farmers. Nearly 40% of the total pig population of India is present in the north-eastern (NE) states where pig husbandry plays an important role in the socio-economic development. Pigs reared in the backyards are the only source of livelihood for a majority of poor tribal population in the region. Hardly any CSF vaccination is currently being undertaken in the unorganized pig farming in the NE region due to economic reasons and vaccine unavailability. A thorough understanding of the current epidemiological status of CSF is essential for the effective control of the disease in the NE region. Hence, we carried out molecular characterization of CSFV isolates from field outbreaks during 2011-2012 in the entire north-eastern region of India to establish the genetic groups of prevalent CSF viruses in the region. A total of 17 CSFV isolates obtained from different parts of the NE region were characterized by comparing the sequences of three partial genomic regions of the virus, that is 150 nt of 5' UTR, 190 nt of E2 and 409 nt of NS5B. Of the 17 CSFV isolates, 15 isolates belonged to 1.1 (88.2%) and two isolates (11.8%) belonged to 2.2 subgenogroup. The genogroup 2.2 CSFV were associated with outbreaks in Arunachal Pradesh that shares international borders with Bhutan, Myanmar and China. Genogroup 2.2 CSFV isolated in the present study shared high level of sequence similarity with 2.2 viruses form China, raising the possibility of virus incursion from this region. In summary, we found a continued predominance of 1.1 subgroup and an emergence of 2.2 subgroup CSFV in NE region of India.

Detection of Shiga toxin-producing Escherichia coli and enteropathogenic Escherichia coli in faecal samples of healthy mithun (Bos frontalis) by multiplex polymerase chain reaction.

Faecal samples obtained from 190 healthy mithuns were examined for the presence of Escherichia coli. Total one-hundred and five E. coli isolates were obtained from these samples, which belonged to 55 different serogroups. These isolates were subjected to multiplex polymerase chain reaction (m-PCR) for detection of stx1, stx2, eaeA and hlyA genes. Twenty-three (21.90%) E. coli isolates belonging to 14 serogroups revealed the presence of at least one virulence gene when examined by m-PCR. Nineteen percent and 2.85% of the mithuns were found to carry Shiga toxin-producing E. coli (STEC) and enteropathogenic E. coli, respectively. stx1 and stx2 genes were found to be prevalent in 7 (6.67%) and 18 (17.14%) of the isolates respectively, whereas eaeA and hlyA genes were found to be carried by three (2.85% each) isolates. Interestingly, none of the STEC isolates belonged to serogroup O157.

Detection of heat-stable and heat-labile enterotoxin genes of Escherichia coli in diarrhoeic faecal samples of mithun (Bos frontalis) calves by polymerase chain reaction.

AIMS: To find out the prevalence of different serogroups of Escherichia coli (E. coli) and to detect heat-stable (ST) and heat-labile (LT) enterotoxin genes of enterotoxigenic E. coli (ETEC) from the faeces of mithun calves with diarrhoea. METHODS AND RESULTS: Faecal samples obtained from 65 diarrhoeic mithun calves of under 2 months of age were examined for E. coli using polymerase chain reaction (PCR). Fifty-four E. coli isolates were obtained from those samples, which belonged to 38 different serogroups. Out of 54 isolates tested by PCR, two isolates (3.70%) belonging to serogroups O26 and O55 were found to possess gene that code for ST enterotoxin and one isolate (1.85%) belonging to serogroup O125 was found to carry LT enterotoxin gene. CONCLUSIONS: Escherichia coli isolates from diarrhoeic mithun calves were found to possess ST and LT enterotoxin genes, which are designated as ETEC, and these isolates can be detected through PCR using specific primers. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the isolation of ETEC possessing ST and LT enterotoxin genes for the first time and ETEC could be a cause of diarrhoea in mithun calves leading to calf mortality.

Seroprevalence of Toxoplasma gondii antibodies in free-ranging mithuns (Bos frontalis) from India.

Studies conducted on free-ranging mithuns of Nagaland, India revealed that the overall seroprevalence of antibodies to Toxoplasma gondii in mithun was 42% (95% CI = 33-51) when detected by modified direct agglutination test. Highest (prevalence = 57%, 95% CI = 43-71) seroprevalence was found in mithuns above 3 years of age and the lowest (prevalence = 18%, 95% CI =4-32) in mithuns of 1-2 years old. No statistically significant difference was observed between male (prevalence = 40%, 95% CI = 26-54) and female (prevalence = 44%, 95% CI = 31-57) seroprevalences. The study also revealed that the maximum number (57%) of animals with the highest titre (1 : 3200) were above 3 years of age. This is the first serological survey for Toxoplasma gondii antibodies in free-ranging mithuns from India.

Bluetongue infection rate in mithun (Bos frontalis) in the north-eastern upland region of India.

Studies conducted on 106 mithun at the National Research Centre on Mithun and 66 free-ranging mithun in Nagaland, India, revealed an infection rate with bluetongue virus of 86%, using a commercially available competitive enzyme-linked immunosorbent assay. Animals were grouped according to their age: 36 of 1 to 2 years of age, 50 of 2 to 4 years of age and 86 aged 4 years and over. The highest infection rate (98%) was found in mithun > 4 years old and the lowest (58%) in those 1 to 2 years old. No statistically significant difference was observed between infection rates of males (89%) and females (85%). The infection rate was higher (95%) in free-ranging mithun than in mithun kept under a semi-intensive system (80%). This is the first report of serological evidence of antibodies to bluetongue virus in mithun. The possible role of vectors in the epidemiology of bluetongue virus infection in mithun is discussed briefly.

Serological evidence of Neospora caninum infection in mithun (Bos frontalis) from India.

Studies conducted on mithuns maintained at National Research Centre on mithun, Indian Council of Agricultural Research (ICAR), Nagaland, India and mithuns found in free-ranging condition of Nagaland revealed that the overall prevalence of antibodies to Neospora caninum in mithun was 10% (95% CI=5-15) when detected by a commercially available competitive enzyme-linked immunosorbent assay test. Highest (prevalence rate=16, 95% CI=8-24) seroprevalence was found in mithuns above 3 years of age and lowest (prevalence rate=2, 95% CI=0-6) in mithuns of 2-12 months old. No statistically significant difference was observed between male (prevalence rate=7, 95% CI=0-14) and female (prevalence rate=12, 95% CI=6-18) seroprevalences. The seroprevalence was found to be higher (prevalence rate=20, 95% CI=9-31) in mithuns found in free-ranging condition in comparison to mithuns kept in semi-intensive system (prevalence rate=5, 95% CI=1-9). This is probably the first report on serological evidence of N. caninum infection in mithun. The possible role of sylvatic fauna in the epidemiology of N. caninum infection mithun is also discussed in brief.

Serological evidence of coronavirus infection in mithuns (Bos frontalis) from India.

Studies conducted on mithuns at the National Research Centre on Mithun, and mithuns found in free-ranging conditions in Nagaland, India, revealed that the overall prevalence of antibodies to coronavirus was 80% (95% confidence interval [CI] 77-83) when detected by a commercially available enzyme-linked immunosorbent assay. The highest seroprevalence (92%; 95% CI 89-95) was found in mithuns above three years of age, and the lowest (53%; 95% CI 36-70) in mithuns of 7 to 12 months old (P = 0.00005). The study also revealed a higher degree of positivity (++++) in mithuns above three years old than in those below one year. No statistically significant difference in seroprevalence was observed between male (80%; 95% CI 75-85) and female mithuns (80%; 95% CI 76-84). The seroprevalence was found to be higher (92%; 95% CI 89-95) in mithuns reared in semi-intensive systems compared with those kept under free-ranging conditions (56%; 95% CI 49-63; P = 0.003). This is probably the first serological evidence of coronavirus infection in mithuns. The possible role of adults in the epidemiology of coronavirus infection in mithuns is also discussed.

Prevalence of Cryptosporidium parvum in mithuns (Bos frontalis) from India.

Systematic examination of faecal samples from mithuns maintained at the National Research Centre on Mithun, Indian Council of Agricultural Research (ICAR), Nagaland, India and mithuns found in free-ranging condition of Nagaland, by using a commercially available enzyme-linked immunosorbent assay kit, revealed that the overall prevalence of Cryptosporidium parvum in mithun was 56% (95% CI=48-64). Highest (prevalence rate=81, 95% CI=67-95) prevalence was found in mithuns of 1-6 months of age and lowest (prevalence rate=42, 95% CI=31-53) in mithuns above 2 years of age. The prevalence was found to be higher (94%) in diarrhoeic animals in comparison to non-diarrhoeic group (51%). No statistically significant difference was observed between male (prevalence rate=61, 95% CI=48-74) and female (prevalence rate=53, 95% CI=43-63) prevalences. The prevalence was found to be higher (prevalence rate=64, 95% CI=55-73) in mithuns kept in semi-intensive system than mithuns found in free-ranging condition (prevalence rate=40, 95% CI=27-53). The risk factor that may play a pivotal role in the epidemiology of cryptosporidiosis in mithun is also discussed in brief. The zoonotic implication of the disease is also discussed. This is probably the first report on prevalence of Cryptosporidium parvum in mithun.

Seroprevalence of Toxoplasma gondii antibodies in captive mithuns (Bos frontalis) from India.

One hundred and four mithuns from India were tested for serum antibodies to Toxoplasma gondii by modified direct agglutination test. The overall prevalence of T. gondii antibodies in mithun was 28% (95% CI=19-37). Highest (prevalence rate=46, 95% CI=30-62) seroprevalence of T. gondii antibodies was found in mithuns above 3 years of age and lowest (prevalence rate=14, 95% CI=1-27) in mithuns of 6 months-1 year old. No statistically significant difference was observed between male (prevalence rate=23, 95% CI=10-36) and female (prevalence rate=31, 95% CI=20-42) seroprevalences (P=0.53). Significant difference was observed in the seroprevalence of T. gondii antibodies in mithuns of different strains with the highest (48%) seroprevalence recorded in Arunachal and lowest (14%) in Mizoram strain. The seroprevalence was higher in farm located at lower altitude (305 m a.s.l.) than the farm located at higher altitude (2134 m a.s.l.). This is the first serologic survey for T. gondii antibodies performed on mithuns from India.