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Molecular and pathological studies on natural cases of bovine theileriosis.

ABSTRACT: The present communication is a part of study conducted on 32 cases of bovine lymphadenopathies. Out of which, six cases of bovine theileriosis were diagnosed on the basis of peripheral blood smear examination and gross lesions in lymph nodes and visceral organs. Nested PCR using two primer sets N516/N517 and 14136/294 was conducted on samples collected from affected lymph nodes confirms Theileria annulata infection in five out of six cases. Sequencing analysis of amplified product showed 92, 94 and 93 % homology of isolate TH1_Bovine, TH2_Bovine and THEN_Bovine for T. annulata with T. annulata Tamilnadu and Pant Nagar. An upregulation of Th2 cytokines in the cases of theileriosis was observed as the level of TNF-? in individual animals varies from higher value (1028 pg/100 ?g protein) to as low as 500 pg/100 ?g protein. An increase in level of Th2 cytokines IL-4 and IL-10 was also observed. The present study concluded that pathological studies and cytokine analysis of lymph nodes are of paramount importance in disease diagnosis and associated immune response of the animal with lymphadenopathies.

SUBMITTER: Gupta A

PROVIDER: S-EPMC5339203 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Molecular and pathological studies on natural cases of bovine theileriosis.

Gupta Anamika A Gupta Kuldip K Leishangthem Geeta Devi GD Bal Mandeep Singh MS Sood N K NK Singh Amarjit A

Journal of parasitic diseases : official organ of the Indian Society for Parasitology 20160526 1

The present communication is a part of study conducted on 32 cases of bovine lymphadenopathies. Out of which, six cases of bovine theileriosis were diagnosed on the basis of peripheral blood smear examination and gross lesions in lymph nodes and visceral organs. Nested PCR using two primer sets N516/N517 and 14136/294 was conducted on samples collected from affected lymph nodes confirms <i>Theileria annulata</i> infection in five out of six cases. Sequencing analysis of amplified product showed ...[more]

PMID: 28316415

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Project description:BACKGROUND: Despite its global recognition as a ruminant pathogen, cases of Chlamydia pecorum infection in Australian livestock are poorly documented. In this report, a C. pecorum specific Multi Locus Sequence Analysis scheme was used to characterise the C. pecorum strains implicated in two cases of sporadic bovine encephalomyelitis confirmed by necropsy, histopathology and immunohistochemistry. This report provides the first molecular evidence for the presence of mixed infections of C. pecorum strains in Australian cattle. CASE PRESENTATION: Affected animals were two markedly depressed, dehydrated and blind calves, 12 and 16 weeks old. The calves were euthanized and necropsied. In one calf, a severe fibrinous polyserositis was noted with excess joint fluid in all joints whereas in the other, no significant lesions were seen. No gross abnormalities were noted in the brain of either calf. Histopathological lesions seen in both calves included: multifocal, severe, subacute meningoencephalitis with vasculitis, fibrinocellular thrombosis and malacia; diffuse, mild, acute interstitial pneumonia; and diffuse, subacute epicarditis, severe in the calf with gross serositis. Immunohistochemical labelling of chlamydial antigen in brain, spleen and lung from the two affected calves and brain from two archived cases, localised the antigen to the cytoplasm of endothelium, mesothelium and macrophages. C. pecorum specific qPCR, showed dissemination of the pathogen to multiple organs. Phylogenetic comparisons with other C. pecorum bovine strains from Australia, Europe and the USA revealed the presence of two genetically distinct sequence types (ST). The predominant ST detected in the brain, heart, lung and liver of both calves was identical to the C. pecorum ST previously described in cases of SBE. A second ST detected in an ileal tissue sample from one of the calves, clustered with previously typed faecal bovine isolates. CONCLUSION: This report provides the first data to suggest that identical C. pecorum STs may be associated with SBE in geographically separated countries and that these may be distinct from those found in the gastrointestinal tract. This report provides a platform for further investigations into SBE and for understanding the genetic relationships that exist between C. pecorum strains detected in association with other infectious diseases in livestock.

Project description:Toxocara vitulorum is an Ascarid nematode infecting the small intestine of buffalo and cattle particularly neonate calves, with the postnatal route through milk is the main infection source. However, little is known about shedding rates and the optimum detection methods of T. vitulorum larvae in the milk of the infected bovine hosts. In this study, we aimed to evaluate the use of two methods, microscopy and PCR, and their detection limits both under the experimental and natural infection situations. In doing this, T. vitulorum eggs extracted from naturally occurring adult female worms were successfully subjected to experimental embryonation, and larvae were implemented in experimental infection of milk in ascending infection doses of 0, 1, 5, 10, 20, 50 larvae/2-ml milk samples. With the except of negative control, microscopy-based examination detected larvae in all samples, albeit with means, ranges, and the total number of larvae were detected in exponential rates relative to larvae densities in milk samples. PCR technique corresponded well to microscopy in detecting genomic DNA of T. vitulorum larvae in all milk samples down to a single larva/sample. On the other hand, and by applying the same methodology approach on 50 naturally-occurring bovine colostrum/milk samples, 13 (26%) and 20 (40%) samples were tested positive for T. vitulorum infection by microscopy and the PCR-based detection, respectively. Of these, 11 out of 26 buffalo samples (42.30%) and 2 out of 24 cow samples (8.33%) were tested positive by microscopy, while 16 (61.54%) and 3 (12.50%) of buffalo and cow samples were tested positive by PCR, respectively. By applying the Agreement Coefficient, substantial agreement (0.77) between molecular and microscopy detection was detected from all tested samples. In conclusion, larvae of T. vitulorum were unequivocally detected by microscopy and molecular methods in milk samples both under the experimental and natural field situations. Nevertheless, slightly higher rates by PCR than microscopy were obtained when detecting naturally-infected milk samples. To the best of our knowledge, this is the first in situ detection of larvae of T. vitulorum in the milk of the naturally infected animals.

Dataset Information

Molecular and pathological studies on natural cases of bovine theileriosis.

Publications

Molecular and pathological studies on natural cases of bovine theileriosis.

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