The interplay of cytokines in bovine tropical theileriosis: a mini review.

Studying cytokine profiling in Theleria annulata infection enhances our understanding of how the immune response unfolds, the intricate interactions between the host and the parasite, the strategies employed by the parasite to evade the immune system, and potential avenues for developing treatments. The generation of pro-inflammatory cytokines plays a pivotal role in the immune response against T. annulata infection. Elevated concentrations of these cytokines potentially contribute to the manifestation of clinical symptoms associated with the disease, such as fever, anemia, exophthalmia, and weight loss. The production of anti-inflammatory cytokines potentially serves as a regulatory mechanism for the immune response, preventing the development of severe disease. Nevertheless, in animals afflicted by T. annulata infection, there is often a notable decrease in the levels of these cytokines, suggesting that they may not be as effective in mitigating the disease as they are in uninfected animals. This knowledge can be harnessed to develop improved diagnostic methods, treatments, and vaccines for tropical theileriosis. The objective of this current mini review is to achieve the same goal by consolidating the available knowledge of cytokine interactions in Bovine Tropical Theileriosis (BTT).

Phylogenetics of Sarcocystis fusiformis isolates based on 18S rRNA and cox 1 genes.

Sarcocystosis is a significant meat borne coccidian disease with immense zoonotic potential. Sarcocystis fusiformis is the most prevalent Sarcocystis spp. affecting buffaloes across the globe. Most of the molecular characterization works on S. fusiformis are from Egypt and there is no record of such work from India. In the present study, 21 isolates of S. fusiformis from Northern India were characterized for 18S rRNA (MF595821-MF595841) and cox 1 (MF423105-MF423119 and MH899162-MH899167) genes. S. fusiformis was seen as a monophyletic sister group to S. cafferi on the phylogenetic tree comprising of different Sarcocystis spp. Both genes placed S. fusiformis close to those Sarcocystis spp. which have felids as definitive hosts in comparison to those with canids as definitive host. A total of 15 and 7 haplotypes were noticed for both the genes, respectively. The studied Indian isolates showed 99.1-100.0% and 99.2-100.0% nucleotide homologies within themselves for both the respective gene loci. Over all, cox 1 gene was found to be better in delineating the evolutionary phylogenetics in comparison to 18S rRNA gene. The findings are important from evolutionary point of view.

Epidemiology, haematology and molecular characterization of haemoprotozoon and rickettsial organisms causing infections in cattle of Jammu region, North India.

BACKGROUND: The present study was aimed at establishing the prevalence, epidemiology and molecular characterization of major haemoprotozoons (Babesia and Theileria) and rickettsia (Anaplasma) of cattle in Jammu region (North India) using microscopy and Polymerase Chain Reaction (PCR). Hematology, microscopy and PCR based prevalence studies were undertaken with 278 whole blood samples from cattle. Molecular prevalence studies were followed by genetic characterization of the isolates of Babesia, Anaplasma and Theileria spp. based on 18S rRNA, 16S rRNA and Tams1 gene, respectively. The data related to metrology and epidemiological variables like temperature, rainfall, season, age and type of livestock rearing was analyzed and correlated with occurrence of disease by statistical methods. RESULTS: The prevalence based on microscopy was 12.9% (36/278) whereas PCR recorded 30.22% (84/278) animals positive for haemoparasitic infections. All the samples found positive by microscopy were also recorded positive by PCR. Thus the study revealed prevalence of Babesia bigemina, Anaplasma marginale and Theileria annulata to be 9.7, 16.5 and 0.7% respectively. The metrological and epidemiological variables made inroads for the propagation of vector ticks and occurrence of infection. Haematological alterations predominantly related to decrease in haemoglobin, red blood cell count and packed cell volume were evident in diseased animals and collaterally affected the productivity. Further the genetic characterization of Babesia bigemina. (MN566925.1, MN567603, MN566924.1), Anaplasma marginale. (MH733242.1, MN567602.1) and Theileria annulata (MT113479) provided a representative data of the isolates circulating in the region and their proximity with available sequences across the world. CONCLUSIONS: Despite holding much significance to the animal sector, comprehensive disease mapping has yet not been undertaken in several parts of India. The present study provides a blue print of disease mapping, epidemiological correlations and genomic diversity of Babesia bigemina, Anaplasma marginale and Theileria annulata circulating in the region.

The phenotypic and haemato-biochemical appraisal of tropical theileriosis in newborn calves.

Tropical theileriosis is one of the major causes of newborn calves mortality. Observation of clinical manifestations is important while making the presumptive/tentative diagnosis of tropical theileriosis in newborn calves. The phenotypic and haemato-biochemical appraisals of tropical theileriosis could be of great help to make a holistic therapeutic plan for diseased newborn calves. Therefore, the present study aimed to evaluate the haemato-biochemical and phenotypic diagnostic markers of tropical theileriosis in newborn calves. A total of 43 newborn calves naturally infected with Theileria annulata and 16 age-matched healthy calves were enrolled. The percentage distribution of clinical markers was generalized lymph nodes enlargement (100%), pyrexia (97.67%), respiratory distress (95.34%), tick infestation (90.69%), anorexia (88.37%), pica (81.39%), pallor mucous membrane (67.44%), hyperlacrimation (58.13%) and exophthalmia (30.22%). Haemograms including TEC, Hb and HCT were found to be significantly (P ≤ 0.001) lowered in diseased calves. Remarkable alterations in the leukogram panels were not observed. Serum glucose, total protein, albumin and globulin concentrations of calves with theileriosis were significantly (P ≤ 0.001) lower than healthy ones, whereas triglycerides and total cholesterol levels of diseased calves were significantly (P ≤ 0.001) higher. Significantly (P ≤ 0.001) elevated activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) enzymes were observed in diseased calves. An evaluation of clinical phenotypes could be helpful to initiate quick treatment of diseased calves in field conditions and save the lives of sick calves of economically poor farmers. Altered haemato-biochemical panels to be appraised by veterinary clinicians while making a therapeutic plan of tropical theileriosis.

Role of cytokines in the clinical manifestation of exophthalmia in newborn calves with tropical theileriosis.

The present study aimed to evaluate the pathology of the exophthalmia and the host-immune response in naturally Theileria annulata-infected calves. The newborn calves detected positive for theileriosis were grouped into calves with theileriosis and absence of exophthalmia (n = 30), and calves with theileriosis and the presence of exophthalmia (n = 13). Sixteen healthy calves, free from any haemoprotozoal infection, were kept as healthy controls. A significantly (P ≤ .001) higher circulating levels of tumour necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were estimated in diseased calves with and without exophthalmia as compared to healthy controls. Contrarily, significantly (P ≤ .01) lower interferon-γ (IFN-γ) level was estimated in diseased calves. The diseased calves with exophthalmia revealed significantly higher levels of TNF-α (P ≤ .001) and IL-10 (P ≤ .006) as compared to the diseased calves without exophthalmia. The diseased calves were not found to have an elevated intraocular pressure; rather they had significantly (P ≤ .001) lower intraocular pressure compared to the healthy controls. An elevated systemic TNF-α level might be attributed to the exophthalmia in calves with tropical theileriosis. The elevated circulatory IL-10 and reduced IFN-γ levels could be one of the strategies of Theileria annulata to escape the host immunity.

Phylogenetic characterization of Setaria equina and its association with other filarids.

Molecular characterization studies on Setaria equina are limited. The present study aimed to characterize S. equina at the cytochrome c oxidase gene and to examine its phylogenetic relationships with other filarid species. Sequence analysis showed 100% nucleotide homology with an S. equina sequence from Italy (AJ544873). However, both sequences exhibited 7 nucleotide substitutions from a S. equina donkey isolate from Egypt (MK541847). Overall, S. equina formed a monophyletic sister group to Setaria tundra. All Setaria spp. examined formed a separate group on the phylogenetic tree that was related to corresponding Onchocerca spp. and Dirofilaria spp. clades. Human filarid worms-Brugia spp. and Wuchereria spp. grouped in a separate clade alongside Theilezia spp. Dipetalonema spp.-formed a separate group at the top of the tree.

Comparison of different PCR protocols and respective primer sets from pool of TAMS 1 gene for diagnosis of calf theileriosis from semi arid India.

The polymorphic nature of Theileria annulata merozoite surface antigen (TAMS 1) attributes to limitation in PCR based detection of various T. annulata genotypes present in different geographical domains across the globe. Multiple reports of failure of detection of tropical theileriosis using classical N516/517 primer set in the studied area were noticed. Hence, three single PCR protocols using N516/517, TAMS F/R and NTA F/R primer sets encoding different portions of TAMS 1 gene and two nested protocols, using combinations of these three primers, were compared to find out the most suitable primer set for diagnosis of calf theileriosis in studied area. The studied area constitutes the semi-arid theileriosis endemic area of Northern India. The various PCR protocols were tested on 75 clinically confirmed cases of calf theileriosis. Alongside, 25 confirmed theileriosis negative blood samples and DNA of other haemoprotozoa were also tested for specificity of these primer sets. Results revealed that the primer set NTA F/R to be more suitable in detecting the circulating T. annulata genotypes in the studied area in comparison to the classical N516/517 primer set. None of the primers gave false positive amplification with negative samples and/or DNA of other haemoprotozoa.

First report of molecular characterization and phylogenetic analysis of Sarcocystis tenella from India.

Sarcocystis tenella is a common tissue coccidian parasite of sheep. It is reported worldwide with high prevalence rate ranging from 9 to 100%. However, there are very limited reports of this parasite from the Indian context and those reports are totally based on the morphology alone. When it comes to molecular characterization, such studies are absent from India. The present communication reports the first characterization study of S. tenella from India. 18S rRNA ribosomal gene and mitochondrial cytochrome c oxidase subunit I (cox1) genes were used for molecular characterization and phylogenetic analysis alongside standard histopathology of sarcocysts. Five Indian isolates were characterized for each gene, and respective sequences were submitted in the NCBI. Two haplotypes were noticed, both for the 18S rRNA and cox1 gene showing 99.8-100.0% and 99.7-100.0% nucleotide homologies within themselves, respectively. When compared with other sequences of S. tenella across the globe, the present isolates showed 93.3-99.9% nucleotide homology based on 18S rRNA gene and 95.2-99.8% nucleotide homology based on cox1 gene, respectively. In both the 18S and cox1 phylogenetic trees, respective sequences of S. tenella were placed with monophyletic cluster which was sister to a cluster comprising of sequences of S. gracilis and S. alces.

Variation in cardiac markers and electrocardiographic alterations in young calves naturally infected with bovine tropical theileriosis.

The present study was designed to assess the deleterious effects of bovine tropical theileriosis on the cardiovascular system and the consequent myocardial involvement in young calves. Myocardial effects in parasitic diseases are often neglected. Hemolytic anemia, associated secondary hypoxia, and vasculitis are cardinal features of bovine theileriosis. In the present study, electrocardiogram (ECG) alongside serum cardiac troponin I (cTnI) and creatinine phosphokinase-myocardial band (CPK-MB) concentrations were analyzed in infected, treated, and control groups of young calves. Non-significant alterations were noticed in ECG. However, certain signs like sinus tachycardia, first-degree AV block, atrial premature complex, left atrial hypertrophy, and right atrial hypertrophy were found on consistent basis in infected calves. A significant increase in the serum concentration levels of cTnI and CPK-MB was noticed in infected calves followed by significant fall in both these biomarkers post treatment. cTnI and CPK-MB can definitely be used as myocardial markers in theileriosis-affected animals.

Standardization and validation of simple PCR, duplex PCR and RAPD in comparison to blood smear examination for diagnosing bovine tropical theileriosis.

Bovine Tropical Theileriosis (BTT) is an important vector-borne protozoan disease that imposing serious constraints on the health and productivity of domestic cattle. It is matter of common fact that following recovery from primary infection, cattle become persistent carriers and act as reservoirs of infection thereby, playing a critical role in disease epidemiology. The present study describes the comparative diagnostic efficiency of simplex PCR, duplex PCR and RAPD assays for detection of Theileria annulata in cattle. An optimized simple PCR and duplex PCR assay were established using TAMS F/R as primer sets encoding for 721 bp amplicon alongside a RAPD with arbitrary primer coding for 963 bp product of T. annulata. The simple PCR and duplex PCR detected pathogen with almost same level of sensitivity, irrespective of whether its DNA was amplified in isolation or together with DNA of another pathogen without nonspecific amplifications. RAPD failed to give comparable results and suffered from limitations of sensitivity as well as specificity. The developed assays may be seen as a good tool for epidemiological studies aiming at assessing the burden of chronic infections and improving control of the associated diseases in endemic regions.

First report of molecular characterization and phylogenetic analysis of RoTat 1.2 VSG of Trypanosoma evansi from equine isolate.

Rotat 1.2 variant surface glycoprotein (VSG) is considered to be an important VSG expressed in most of the isolates of Trypanosoma evansi. This makes the molecule an important candidate for both molecular- and serological-based detection of surra. There are ample reports of existence of this gene in isolates from cattle, buffalo, and camel across the world. Of late, there are reports of its absence from a fewer isolates of T. evansi of murine and wildlife origin. Search of literature revealed no reports from horses. The present communication presents the first report of molecular cloning and characterization of Rotat 1.2 VSG from horse isolate of T. evansi from semi-arid region of India. Alongside, the gene was compared with various other isolates across the world. Interestingly, the isolate was found to be closer to camel isolates from Egypt than the other known isolates from India and Kenya.

Serodiagnosis of Toxoplasma gondii infection in bovines from Kerala, India using a recombinant surface antigen 1 ELISA.

Data on the prevalence of toxoplasmosis in farm animals from India is scanty. Though a few reports exist on prevalence of toxoplasmosis in small ruminants, information on toxoplasmosis in large ruminants is virtually nonexistent from India. An antibody detection recombinant ELISA specific for Toxoplasma gondii was laboratory standardized using recombinant surface antigen 1 (SAG1) protein. A 958-bp truncated sequence coding for tachyzoite stage specific SAG1 protein was amplified and expressed in Escherichia coli BL21(DE3) cells. A high-level expression of the histidine-tagged thioredoxin fusion protein was obtained after 8 h of incubation. The recombinant protein was affinity purified by Ni-NTA agarose chromatography and characterized by SDS-PAGE and Western blot. Subsequently, the diagnostic potential of the recombinant protein was assessed with 258 cattle sera samples from field by a laboratory standardized recSAG1 ELISA. Sera from 71.8% of the cattle showed sero positivity for T. gondii specific IgG. The sensitivity and specificity of the recSAG1 ELISA were 84.38% and 87.88%, respectively in comparison to indirect fluorescent antibody test (IFAT). This is the first report on sensitive serodetection of Toxoplasma infection in bovines from India.

In vitro and in vivo efficacy of extracts of leaves of Eucalyptus globulus on ovine gastrointestinal nematodes.

The rapid development of anthelminthic resistance has limited the success of traditional control programmes in several countries, thereby forcing the researchers to search for alternatives. In vitro anthelmintic activities of crude aqueous and hydro-alcoholic extracts of the leaves of Eucalyptus globulus were investigated against the egg and larvae of naturally infected sheep. In the phytochemical analyses, tannins and flavonoids were the main metabolites identified in the extract. The aqueous extract of E. globulus was also investigated for in vivo anthelmintic activity in naturally infected sheep. The various blood parameters, coupled with effects on marker enzymes and antioxidant status, were evaluated during the trial period. Methanolic extract showed better ED50 (3.756 mg/ml) and ED99 (33.809 mg/ml) values in comparison with aqueous extract (ED50 = 1.502 and ED99 = 7.10 mg/ml) in the egg hatch assay. Inverse was true in larval development and larval paralysis tests. The aqueous extract's ED50 = 19.994 and ED99 = 108.931 mg/ml values in the larval development test and ED50 = 19.994 and ED99 = 108.931 mg/ml in the larval paralysis test were more potent than those of its methanolic counterpart with ED50 = 15.595 and ED99 = 94.493 mg/ml and ED50 = 15.595 and ED99 = 94.493 mg/ml, respectively. A significant amount of 66% faecal egg count reduction was observed in in vivo trail using the aqueous extract on day 21 post treatment, although in initial stages it showed 58.0 and 80% effectiveness on days 7 and 14 post treatment. Though the FCER reduction was somewhat lower in terms of comparison with albendazole, nevertheless, significant and prolong reduction was noticed. No deleterious ill effect was found in any of the haematological and biochemical parameters suggesting that the plant could be safer for use in sheep. Though significant changes were observed in SGPT, RBCs, Hb and RDWc levels, other parameters showed nonsignificant variations within the normal range in the stipulated time of of herbal trial period. Based on the results of the present study, it could be very well concluded that leaves of E. globulus possess good level of anthelminthic efficacy; further research is thereby warranted before recommending it for use in nematode control programme in ovines.

A duplex PCR-based assay for simultaneous detection of Trypanosoma evansi and Theileria annulata infections in water buffaloes.

Trypanosomosis and bovine tropical theileriosis are important vector-borne protozoan diseases imposing some of the serious constraints on the health and productivity of domestic cattle in tropical and subtropical regions of the world. Following recovery from primary infection of both these conditions, animals become persistent carriers and act as reservoirs of infection thereby playing a critical role in disease epidemiology. The present study describes development and evaluation of duplex polymerase chain reaction (PCR) assays for simultaneous detection of Trypanosoma evansi and Theileria annulata in buffaloes. Following in silico screening for candidate target genes representing each of the pathogens, an optimized duplex PCR assay was established using TBR F/R and TAMS F/R as primer sets encoding for products of 164 and 721 bp for T. evansi and T. annulata, respectively. The results were compared and correlated with conventional Giemsa-stained thin blood smear examination and the single PCR assay. The duplex PCR detected each pathogen with the same level of sensitivity, irrespective of whether its DNA was amplified in isolation or together with DNA of another pathogen. Moreover, single and duplex PCRs were able to detect each species with equal sensitivity in serially diluted DNA representing mixtures of T. evansi and T. annulata, and no evidence of nonspecific amplification from nontarget species was observed. The developed assay may be seen as a good tool for epidemiological studies aiming at assessing the burden of dual infections and improving control of the associated diseases in endemic regions.

First molecular evidence of the transplacental transmission of Theileria annulata.

Bovine tropical theileriosis (BTT) is a serious hindrance in the cattle upgradation programme using the exotic germplasm. There is a wide range of variations in the pathobiology alongside clinical symptoms of the animals suffering from BTT. The present paper communicates the first report about the transplacental transmission of T. annulata in a cross bred 2-day old calf. T. sergenti, T. lestoquardi and T. equi are known to undergo transplacental transmission, but baring a single citation in literature, there are no records about the transplacental transmission of T. annulata.

Detection of antibodies to Toxoplasma gondii in domesticated ruminants by recombinant truncated SAG2 enzyme-linked immunosorbent assay.

An antibody detection recombinant enzyme-linked immunosorbent assay (ELISA) specific for Toxoplasma gondii was laboratory standardized using recombinant truncated surface antigen 2 (SAG2) protein of T. gondii. A 483-bp sequence coding for truncated tachyzoite stage-specific SAG2 protein was amplified and ligated in pPROExHT-b expression vector to transform Escherichia coli DH5α cells. A high-level expression of the histidine-tagged fusion protein was obtained after 8 h of incubation. The recombinant protein was affinity purified using Ni-NTA agarose column and characterized by SDS-PAGE and Western blot analysis. Subsequently, the diagnostic potential of the recombinant protein was assessed with 168 field sera samples from sheep, goats and cattle. Among the small ruminants, 50% (n = 60) sheep sera samples and 41.26% (n = 63) goat samples were detected positive for T. gondii-specific antibodies. As far as seroprevalence of toxoplasmosis in cattle is concerned, 64.44% (n = 45) of sera samples assayed were found to be positive. When compared to indirect fluorescent antibody test (IFAT), the sensitivity of the recombinant truncated SAG2 antigen-based ELISA (rec-SAG2-ELISA) ranged from 81.25 to 87.10% while the specificity was 85.71 to 91.43% with substantial agreement between the tests.

Prevalence of haemoparasites infection in diseased bovine and canine of Bhatinda district of Punjab, India.

Infections caused by tick-borne haemoparasites pose a significant global threat to both human and animal health. Within this category, various haemoparasites species belonging to genera like Anaplasma sp., Babesia sp., Ehrlichia sp., Hepatozoon sp., and Theileria sp., are particularly concerning due to their ability to cause diseases in a wide range of hosts, including mammals, birds, and reptiles. The present cross-sectional study involving 580 animals provides annual insights into the prevalence of major haemoparasites infections in the Bathinda region of Punjab. The observed trends indicate that haemoparasites infections were most common in cattle, followed by buffalo and canines. Risk factor analysis revealed that crossbreed cattle were more susceptible to infection, with a prevalence of 35.73% (95% CI 4.28-45.17). Amongst the cattle, adults exhibited a higher vulnerability to haemoparasites infections, with a prevalence of 35.89% (95% CI 5.50-33.64). Conversely, companion animals showed the opposite pattern, with a prevalence of 18.18% (95% CI 9.11-169.27). Furthermore, female dogs had a higher risk of haemoparasites infection, with a prevalence of 16.28% (95% CI 8.36-218.7). In light of these findings, it is imperative to emphasize early diagnosis, prompt antiprotozoals drug treatment, and effective control of tick vectors for the successful recovery of animals afflicted by haemoparasites infections.

Comprehensive genetic diversity and molecular evolutionary analysis of Theileria annulata isolates based on TAMS 1 gene.

Molecular epidemiological studies related to the phylogenetic characterization of Theileria annulata are important in delineating the evolutionary history of the parasite. In the current study, the Theileria annulata (T. annulata) merozoite surface antigen 1 (TAMS 1) gene from 14 bovine isolates of T. annulata originating from semi-arid zone of northern India were amplified and sequenced. TAMS 1 gene sequences (n= 337) reported from 16 countries were subsequently analyzed for haplotype network along with genetic diversity. A total of five haplotypes out of the 14 sequenced isolates and 92 haplotypes out of 337 worldwide sequences are documented in this study. Phylogenetic and molecular evolutionary analyses based on TAMS 1 gene sequences showed that T. annulata is dissipated across different countries and numerous strains are closely linked, even though they belong to different geographical locations. The nucleotide homology between 14 isolates from northern India varied between 91.3 and 100%, whereas it was between 31.5 and 100% when sequences across the globe were compared. Haplotype 14 was recognized as most widely distributed haplotype, with 46 isolates circulating in 10 countries. Globally, negligible genetic distance (FST˂0.15) and very high gene flow (Nm˃1) was found in the five populations of the world (South Asia, East Asia, West Asia, Europe and Africa), supporting the absence of clearly defined subgroups in the phylogenetic analysis. Significant negative values of neutrality tests; Tajima's D (D) and Fu and Li's F (F) provided evidence for recent population expansion through positive selection of advantageous variations.

Phylogenetic characterization and analysis of Sarcocystis buffalonis.

Despite its frequent presence in buffaloes, Sarcocystis buffalonis remains as one of the most under studied parasite. In the present study, isolates of S. buffalonis from,Mathura, Uttar Pradesh India were characterized for 18S rRNA (MF595842-MF595844), cox 1 (MG792800-MG792802), 28S rRNA (MH793418-MH793420) and ITS 1 (MH793421-MH793423) genes. Analysis revealed multiple haplotypes for each individual gene viz., 18S rRNA (three haplotypes), cox 1 (two haplotypes), 28S rRNA (two haplotypes) and ITS 1 (single haplotype). The studied Indian sequences showed variable homologies for individual gene loci viz., 18S rRNA (99.3-99.9%); cox 1 (99.8-100.0%); 28S rRNA (99.9-100.0%) and ITS 1 (100.0%) The phylogenetic association between S. buffalonis and closely related Sarcocystis spp. infecting buffaloes and cattle was delineated. All these gene loci placed S. buffalonis close to S. hirsuta. The study has generated A vital phylogenetic data about this erstwhile neglected parasite.

An endpoint visualization loop-mediated isothermal amplification (LAMP) for detecting bubaline theileriosis.

Background: Tropical theileriosis is a significant disease affecting the health and production levels of buffaloes in India. It is caused by an apicomplexan-Theileria annulata. The timely and accurate detection of infection is vital for implementing a mass vaccination or control programme in a given area under outbreak. Most of the literature concerned with diagnosis of theileriosis revolves around cattle, and practically, there are very limited assays available for detecting bubaline theileriosis. Loop-mediated isothermal amplification (LAMP) assay certainly amplifies the targeted deoxyribosenucleic acid (DNA) with a comparatively higher efficacy, rapidity and sensitivity. Alongside, minimal use of sophisticated instruments in performing LAMP assay is certainly an add on. The present study describes the application of LAMP assay in diagnosing tropical theileriosis in buffaloes alongside, its comparison with polymerase chain reaction (PCR) and blood microscopical examination. Results: No cross-reaction was seen with DNA of other haemoprotozoan. LAMP was compared with blood microscopy and PCR. LAMP detected infection in 27 out of 100 buffaloes, while blood microscopy and PCR detected disease in 16 and 24 buffaloes, respectively. Conclusion: The sensitivity, specificity and kappa value prediction of LAMP were found to be much higher than the PCR and blood microscopy. The present communication reports the first use of LAMP in detecting theileriosis in buffaloes in the world.