Detection of Antibodies Against Trypanosoma evansi in Sheep by Indirect ELISA in Rayalaseema Region of Andhra Pradesh

The present research was carried out with an objective to improve the diagnostic tools for detection of antibodies against Trypanosoma evansi infection using indirect enzyme-linked immunosorbent assay (ELISA) in sheep. In this study standardized the Indirect ELISA for detection of T. evansi in sheep. The optimum concentration of antigen, test sera and conjugate were determined as 5µg per well, 1:10 and 1: 4000 dilutions, respectively. 464 serum samples were collected from sheep in different parts of the Rayalaseema region of Andhra Pradesh for screening of T. evansi infection. Out of 464 serum samples 46 (9.91%) were found positive by indirect ELISA

Histopathological Observations in Rabbits Experimentally Infected with Trypanosoma evansi

New Zealand white rabbits (n=6) were challenged with the South Indian local strain of Trypanosoma evansi. Each animal was infected with 5x105 trypanosomes subcutaneously. Animals were daily examined for the development of clinical signs using wet blood-films sampled from the ear veins. Clinically, intermittent pyrexia, undulating parasitaemia, anorexia and emaciation were predominant. Three months post infection, rabbits were sacrificed, detailed postmortem examination was carried out and representative tissue samples were fixed. Gross pathological changes including paleness of visceral organs, gelatinization of fat, congested and oedematous lungs,mucoid enteritis, hepatomegaly and splenomegaly were noticed. Histopathologically, internal organs elucidated clear changes consisted of severe hepatic fatty change, pulmonary congestion with thickened interstitial spaces and emphysema, degeneration of the renal epithelium associated with cystic tubular formation and congested red pulp

Efficacy of Enrofloxacin in the Treatment of Recurrent Pyoderma in Dogs

Dogs with a history of more than three episodes of skin infections in a period of one year were selected for a study on recurrent pyoderma. Oral enrofloxacin along with appropriate simultaneous medication for the underlying associated conditions were chosen as therapy for recurrent pyoderma in dogs. Response to therapy was excellent in all the cases. Improvement was noticed by 12 to 20 days and 20 to 26 days in recurrent superficial and deep pyoderma respectively. Relapse occurred in one dog by 45 days due to re-introduction of allergic food. Enrofloxacin proved to be an effective, safe and convenient antibiotic for the treatment of recurrent pyoderma in dogs

Kloniranje, ekspresija i karakterizacija paraflagelarnog gena Rod 2 bičaša Trypanosoma evansi

Paraflagellar rod is the major structural component of the trypanosomatid flagellum and is identified as a complex lattice of filaments which runs parallel to the axoneme throughout most of the flagellar length. The present study was carried out to investigate the existence of the paraflagellar rod (PFR 2) gene in Trypanosoma evansi infecting Indian cattle. Local isolates of T. evansi collected from naturally infected cow were multiplied in Wistar rats. Complementary DNA (cDNA) was synthesized from the RNA of host cell free T. evansi parasites by reverse transcription. The gel purified PCR product (PFR 2 gene of T. evansi) was cloned into the pTZ57R/T vector system. The nucleotide sequence of the PFR 2 gene of the T. evansi S.V.V.U. isolate (Accession No. KT277497) obtained in the present study revealed 100% homology with T. evansi China isolate and 99% homology with T. evansi Izatnagar and Bikaner isolates. The recombinant protein was sub-cloned into pET 32a and expressed in the BL21 (DE3) pLysS expression system. The PFR 2 gene of T. evansi S.V.V.U. isolate was further characterized by determination of its protein profile with SDS-PAGE and western blotting. Indirect ELISA was optimized for detection of the specific antibody titre against the recombinant protein of the PFR 2 gene of T. evansi. In the kinetoplastid species the PFR 2 gene is highly conserved. Therefore the PFR 2 gene was suggested as a vaccine candidate, as well as a diagnostic antigen.Paraflagelarni štapić glavna je strukturna komponenta tripanosomskog biča i dio je kompleksa filamenaza koji teku paralelno s aksonemom duž biča. Istraživanje je provedeno kako bi se ispitalo postojanje paraflagelarnog gena Rod 2 (PFR2) u bičaša Trypanosoma evansi koji invadira goveda u Indiji. Lokalni izolat T. evansi prikupljen od prirodno invadiranih krava umnožen je u Wistar štakora. Komplementarna DNA (cDNA) sintetizirana je iz RNA obrnutom transkripcijom iz stanica neinvadiranih nositelja T. evansi parazita. Pročišćeni PCR produkt (gen PFR2 bičaša T. evansi) kloniran je u vektorski sustav pTZ57R/T. Nukleotidna sekvencija gena PFR2 bičaša T. evansi, izolat S.V.V.U. (pristupni broj KT277497) dobivena u ovom istraživanju pokazala je 100 %-tnu sličnost s izolatom T. evansi China i 99 %-tnu s izolatom T. evansi Izatnagar i Bikaner. Rekombinantni protein ponovno je kloniran u sustavu pET 32a i prikazan u sustavu BL21 (DE3) pLysS. Gen PFR2 bičaša T. evansi, izolat S.V.V.U. dalje je karakteriziran određivanjem proteinskog profila metodama SDS-PAGE i Western blotting. Indirektni test ELISA optimiziran je za dokaz titra specifičnih protutijela za rekombinantni protein gena PFR2 bičaša T. evansi. U kinetoplastida gen PFR2 izrazito je očuvan. Stoga bi se gen PFR2 mogao upotrijebiti za cjepivo te kao dijagnostički antigen

Serum biochemical parameters and cytokine profiles associated with natural African trypanosome infections in cattle.

BACKGROUND: Animal African trypanosomiasis (AAT) greatly affects livestock production in sub-Saharan Africa. In Ghana prevalence of AAT is estimated to range between 5 and 50%. Studies have reported serum biochemical aberrations and variability in cytokine profiles in animals during infection. However, information regarding the biochemical parameters and cytokine profiles associated with natural infections are limited. This study was therefore aimed at investigating changes in the levels of serum biochemical parameters and inflammatory cytokines during a natural infection. METHODS: Nested internal transcribed spacer (ITS)-based PCR and sequencing were used to characterise trypanosome infection in cattle at two areas in Ghana (Adidome and Accra) of different endemicities. The cattle were sampled at four to five-week intervals over a period of six months. Levels of serum biochemical parameters, including creatinine, cholesterol, alkaline phosphatase (ALP), alanine aminotransferase (ALT), total bilirubin and total protein and cytokines (interleukin 10, interleukin 4, interleukin 12, interferon gamma and tumor necrosis factor alpha) were measured in serum samples and then compared between infected cattle and uninfected controls. RESULTS: The predominant trypanosome species detected in Accra (non-endemic) and Adidome (endemic) were Trypanosoma theileri and Trypanosoma vivax, respectively. Serum biochemical parameters were similar between infected and uninfected cattle in Accra. Infected cattle at Adidome however, had significantly higher levels of ALP, creatinine, total protein and total bilirubin (P < 0.05) and significantly lower levels of cholesterol (P < 0.05) at specific time points. At basal levels and during infection, significantly higher pro-inflammatory to anti-inflammatory (Th1/Th2) cytokine ratios were observed in cattle at Adidome compared to Accra (P < 0.05), indicating a shift towards Th1 immune response in Adidome. Levels of IL-10 were, however, significantly elevated in infected cattle in Accra (P < 0.05), suggesting high anti-inflammatory cytokine response in Accra. CONCLUSION: These results suggests that cattle in an endemic area repeatedly infected with trypanosomes of different species or different antigenic types demonstrate high pro-inflammatory (Th1) immune response and biochemical alterations whereas cattle in a non-endemic area with predominantly chronic T. theileri infections demonstrate high anti-inflammatory response and no biochemical alterations

MANAGEMENT OF DERMATOLOGICAL LESIONS ASSOCIATED WITH BABESIA GIBSONI IN DOGS

Three dogs were presented to the clinic with the history of recurrent dermatological abnormalities, fever, dyspepsia, lethargy and progressive weight loss. Dermatological findings observed were dry exfoliations, alopecia, hemorrhagic spots over abdomen and groin regions, inter digital ulcerative lesions, hyperkeratosis of digital pads, brittleness of nails and pododermatitis. Microscopic examination of the stained peripheral blood smears revealed the Babesia gibsoni organisms in the erythrocytes. Dogs showed anaemia, leucopaenia, thrombocytopenia, hyper globulinemia and low albumin levels. Uneventful recovery was recorded after treatment with intra muscular administration of two doses of diminazene aceturate @ 7.0 mg/kg body weight and oral administration of clindamycin @ 25 mg/kg body weight twice in a day. Improvement in the clinical findings was noticed by the three days of therapy and complete dermatological clinical cure was obtained after two months of therapy