Bovine cerebral theileriosis: first molecular report in cross bred cattle calf in India.

Bovine tropical theileriosis caused by Theileria annulata, is a serious constraint to Indian dairy industry with more fatal infections in exotic cattle and substantial losses to cross-bred and indigenous zebu cattle. The present communication is to place on record the first report of molecular based confirmed case of cerebral theileriosis caused by T. annulata coupled with its morphological detection, clinical manifestations, haematological alterations and therapeutic management in a cross bred cattle calf from India. After preparation of peripheral thin blood smear from cross bred cattle calf at the site of collection and fixation with methanol, blood sample brought to Department of Veterinary Parasitology, College of Veterinary Science and A.H, Jabalpur and stained by standard protocol for Giemsa staining. Genomic DNA was isolated from the collected blood sample using QIAamp® DNA blood mini kit following the manufacturer's recommendations and PCR was performed. The cross bred cow calf revealed high rise in temperature (105.5°F), increased heart rate, labored breathing with seromucous nasal discharge, enlargement of prescapular lymph node and animal exhibited tonic clonic convulsions in response to any sudden noise. Giemsa stained thin blood smear revealed intraerythrocytic piroplasm and Koch'sblue bodies of T. annulata within the cytoplasm of lymphocytes. The species of Theileria was confirmed by molecular amplification of genomic DNA as T. annulata.

Detection of deltamethrin, cypermethrin and flumethrin efficacy against buffalo lice-Haematopinus tuberculatus.

The cattle and buffalo farm practices have been adopted differently by farmers in India but the infestation of ectoparasites including louse has been advocated in high population of animals across the country. The aim of this study was to identify the louse morphologically and determine the in vitro efficacy of the insecticides deltamethrin, cypermethrin and flumethrin against the buffalo louse, Haematopinus tuberculatus. The present research work was conducted using lice collected from organized buffalo dairy farms of Mhow block, Indore district of Madhya Pradesh, India. The adult's lice were collected from heavily infested regions of the body and tail of buffaloes. Some of the collected adult's lice were preserved for morphological identification in 70% alcohol. Briefly, in vitro treated surface bioassay utilizing a cloth rectangle that allows lice to move freely has been used. The concentrations were prepared as 30, 60, 90 and 120 ppm for deltamethrin and flumethrin, whereas for cypermethrin, 100, 200, 300 and 400 ppm concentrations were prepared in distilled water. The 600 µl of each concentration was spread evenly over a cloth rectangle held in the bottom of a Petri plate. Ten adult lice were used for each concentration in triplicate (n = 30) and the same is maintained for control. The vitality of the louse was assessed at various intervals: 30, 60, 120, 180 and 240 min. The lousicidal efficacy was determined by using in vitro bioassays with deltamethrin, cypermethrin and flumethrin. It is observed that as the concentration of insecticides increases with exposure time, mortality of lice is also increased. The current study reveals that cypermethrin and flumethrin were effective in their recommended doses but in the case of deltamethrin, the lice showed a low level of resistance. Furthermore, this type of study on buffalo louse has not conducted in Mhow region of Madhya Pradesh where heavy infestation of lice occurs on buffalo.

Pubic lice infestation in man from Mhow, Madhya Pradesh.

Worldwide, 2-10% of human population is infested with pubic lice which are mostly found in the hair of pubic area, occasionally under the armpits, in the beard or mustache and on the eyebrows and eyelashes. They are usually transmitted through sexual contact or through toilet seats and bedding material. A twenty-year-old boy suffering from severe itching in the genital region during night hours submitted the lice for their identification in Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Mhow. These lice were identified as Phthirus pubis.

Incidence of amphistome infection in sheep from Mhow, Madhya Pradesh.

A total of 445 faecal samples and 128 rumens of sheep collected from Slaughter house, Mhow (Madhya Pradesh) were examined for a period of 1 year from February 2011 to January 2012. Faecal sample examination based, incidence of amphistomes was found to be 23.37% from the study area. The incidence of amphistomes was significantly higher (p < 0.01) during summer (36.75%) than winter (27.74%) and monsoon (3.52%). Age-wise, non-significantly higher infection was observed in ≥1 year-old animals (23.61%) than <1 year-old animals (23.11%). Non-significantly higher rate of infection was recorded in case of females (25.63%) than that of males (19.64%). Out of 128, 49 (38.28%) rumens were found positive for amphistomes. The prevalence of Paramphistomum cervi (27.34%) was found to be significantly (p < 0.01) higher than Gastrothylax crumenifer (10.94%).

Incidence of gastrointestinal nematodes in goats in and around Mhow, Madhya Pradesh.

Gastrointestinal (GI) parasitism in animals is considered as one of the major problems in India causing recurring economic losses to livestock production system owing to morbidity and mortality. But data pertaining to epidemiology of GI nematodes in goats is meager from Malwa region of Madhya Pradesh. Therefore, the present investigation was carried out to study the incidence of GI nematodes in goats in and around Mhow and accordingly a total of 900 faecal samples of goats were collected from Mhow and its adjoining areas and faecal sample examination revealed the incidence of strongyle, Strongyloides papillosus, Trichuris spp. and mixed infection from the study area. Significantly higher infection of GI nematodes was evidenced in the monsoon season (50.67%) when compared with summer (41.33%) and winter (38.33%) seasons. Significantly higher infection rate was documented in >1 year-old-goats (50.43%) than <1 year-old-goats (19.31%). Non-significantly higher infection was noted in females (46.04%) than their male counterparts (39.50%).

Kinetics of serum immunoglobulin isotype response in experimental bovine tropical fasciolosis.

The present communication reports on the kinetics of immunoglobulin isotype response in Fasciola gigantica infected bovine calves. Fifteen Holstein-Friesian cross-bred male calves were assigned to 3 groups (Gr) of 5 calves each and infected with 4-month (Gr-A) and 16-month-old (Gr-B) F. gigantica metacercariae (n=400), respectively, while Gr-C calves served as uninfected control. Infection was terminated by treating the animals with triclabendazole on 28 weeks post-infection (WPI). Sera were collected on 0, 4, 10 and 14 days post-infection (DPI) and subsequently at weekly interval up to 32 WPI. The immunoglobulin isotype response was analyzed by ELISA, using anion exchange purified antigen fraction. An IgG response against F. gigantica infection was evoked by 3 and 2 WPI in animals of Gr-A and Gr-B, respectively with peak antibody response at 13 WPI. Elicitation of an early IgG1 response by 10 and 14 DPI but a delayed IgG2 response at 6 and 4 WPI, in animals of Gr-A and Gr-B, respectively was recorded. An early IgM response was evoked by 10 and 14 DPI and the level peaked at 13 and 12 WPI, with no detectable level at 21 and 15 WPI in animals of Gr-A and Gr-B, respectively. IgA response was elicited at 4 WPI in both the groups and showed the highest titre at 25 and 27 WPI in animals of Gr-A and Gr-B, respectively. Present study indicated an early and predominant response of IgG1, with concurrent expression of delayed and weak IgG2 in calves experimentally infected with F. gigantica.

Prepatent detection of Fasciola gigantica infection in bovine calves using metacercarial antigen.

Metacercarial antigen of Fasciola gigantica was evaluated for early immunodiagnosis of experimental bovine fasciolosis using ELISA and Western blot. In ELISA, the experimental F. gigantica infection was detected as early as 2 weeks post-infection (WPI). The gradual increasing trend of antibody level was observed from 2 to 7 WPI, followed by a plateau, which was maintained up to 14 WPI. In Western blot, sera from experimentally infected calves recognized one distinct polypeptide of 21 kDa in fractionated metacercarial antigen as early as 10th day post infection. From 2 WPI, more polypeptide bands were reacting. Recognition of these protein bands persisted till the end of the experiment (14 WPI). Cattle sera collected from the field showed 34.5% seroprevalence of fasciolosis by ELISA using MAg. Comparative immunoblot studies of metacercarial antigen with anti-Gigantocotyle explanatum and anti-Paramphistomum epiclitum sera revealed that 21 and 25 kDa polypeptides of metacercarial antigen did not cross-react with any of these sera and appear to be unique to F. gigantica and having the desirable qualities of early and specific immunodiagnosis.

Cathepsin L cysteine proteinase in the diagnosis of bovine Fasciola gigantica infection.

Cathepsin L cysteine proteinase from Fasciola gigantica was evaluated for its potential in the early prepatent detection of this helminth infection in bovine calves. Five cross-bred bovine calves were experimentally infected with 400 metacercariae/calf and evaluated for anti-cathepsin L antibody response. F. gigantica infection in these calves could be detected 4 weeks post-infection using an ELISA, dipstick ELISA and Western blotting with 100% sensitivity. The antigen was also used to detect F. gigantica field infection in cattle, by screening 256 sera of these animals by an ELISA, which demonstrated an overall infection rate of 26.95%. Preliminary studies showed that F. gigantica cathepsin L cysteine proteinase does not cross-react with Paramphistomum epiclitum, Gigantocotyle explanatum and hydatid cyst antigens. However, extensive studies on the cross-reactivity of this antigen with related helminth parasites of cattle and buffaloes are required, before this antigen can be considered suitable for immuno-diagnosis of fasciolosis in these ruminants.