Preparation and characterization of excretory and secretory antigen of Cotylophoron cotylophorum and Gastrothylax crumenifer.

Excretory and secretory antigen of adult Cotylophoron cotylophorum and Gastrothylax crumenifer was prepared and characterized by SDS-PAGE. Adult paramphistomes were collected in PBS from rumen of sheep slaughtered at Perambur and Villivakkam slaughter houses, Chennai, Tamil Nadu. Excretory/Secretory (E/S) antigens of G. crumenifer and C. cotylophorum were prepared by incubating in PBS at 28.3 °C overnight in a shaking incubator and centrifuged. The culture supernatant was used as antigen. The protein concentration of E/S antigens of C. cotylophorum and G. crumenifer ranged from 0.4 to 7.1 mg/ml and 3.3 to 5.9 mg/ml, respectively. SDS-PAGE analysis of E/S antigen of C. cotylophorum revealed two polypeptide bands from the range of 32 and 40 kDa. Whereas, SDS-PAGE analysis of E/S antigen of G. crumenifer revealed only one polypeptide band of 35 kDa. Excretory/secretory antigen may be used for serodiagnosis of paramphistomosis infection in sheep.

Incidence of fur mite infestation in laboratory rodents.

Incidence of fur mites in laboratory rodents is reported. Among mice, infestation was found to be highest in male Swiss Albino mice followed by female Swiss Albino mice, male Balb C mice and female Balb C mice. Infestation in Guinea pigs was 50 %. None of the rats screened were found to be positive for fur mites. Species of fur mites identified were Myocoptesmusculinus in mice and Chirodiscoidescaviae in Guinea pigs. Clinical signs in affected animals are also reported.

Incidence of Onchocerca gibsoni in subcutaneous nodules of cross bred Jersey cows: case report.

Onchocerca gibsoni subcutaneous nodules in four cross bred Jersey cows aged 5-6 years brought for post mortem with calcified and caseated skin nodules in the lateral flank region is reported. Examination and dissection of these nodules revealed that they were "worm nests" of Onchocerca sp. of filarid nematodes. The worm nests were carefully dissected and filarid worms were extracted out. Multiple numbers of worms were recovered from each nodule ranging from 15 to 20. Female worms were found inside the worm nests and were found to be filled with microfilariae. The nodules weighed 2-3 g. Based on the morphological features the worms were identified as O. gibsoni.

A study on the tolerance level of farmers toward human-wildlife conflict in the forest buffer zones of Tamil Nadu.

AIM: The aim of this work was to study the tolerance level of farmers toward different human-wildlife conflict (HWC) situations. MATERIALS AND METHODS: This study was conducted in 24 villages of nine blocks from Kancheepuram, Coimbatore, Erode, and Krishnagiri districts of Tamil Nadu by personally interviewing 240 farmers affected with four different HWC situations such as human-elephant conflict (HEC), human-wild pig conflict (HPC), human-gaur conflict (HGC), and human-monkey conflict (HMC). A scale developed for this purpose was used to find out the tolerance level of the farmers. RESULTS: In general, the majority (61.70%) of the farmers had medium level of tolerance toward HWC, whereas 25.40% and 12.90% belonged to a high and low category, respectively. The mean tolerance level of the farmer's encountering HMC is low (8.77) among the other three wild animal conflicts. In tackling HWC, the majority (55.00%) of the HEC farmers drove the elephant once it entered into their farmland. In the HPC, more than three-fourths of the respondents drove away the wild pig once they were found in farmlands. With regard to the HMC, a less number of them (1.70%) drove the monkey away if monkeys were spotted in their village. With regard to HGC, 95.00% of the respondents frightened the gaurs if their family members were threatened by gaurs. CONCLUSION: The present study suggests that that majority of the farmers had medium level of tolerance toward HWC. The tolerance level of the HMC farmers was lower than other three HWC affected farmers. This study emphasizes the need for necessary training to tackle the problem in an effective manner for wild animal conservation.

Sporulation dynamics of poultry Eimeria oocysts in Chennai.

The infective form of Eimeria is the highly resistant oocyst, which is shed in the faeces of infected animals. Present study was carried out to understand the sporulation dynamics of six Eimeria oocysts viz. E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix and E. tenella in Chennai. Faecal samples of poultry were collected from various poultry farms located in and around Tamil Nadu. Oocysts of various Eimeria species were examined microscopically for sporulation on a 6 h interval basis till complete sporulation is acheived. The sporulation time recorded was 168, 120, 216, 192, 96 and 96 h for E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix and E. tenella respectively. It can be concluded on comparison with previous studies that humid weather conditions delay the sporulation time and dry weather and wet litter is the ideal condition for rapid sporulation.

Occurrence of the pentastomid Porocephalus crotali (Humboldt, 1811) in an Indian rat snake (Ptyas mucosus): a case report.

The occurrence of the pentastomid Porocephalus crotali in an Indian rat snake (Ptyas mucosus) at Chennai, India is reported. The worms were found in the lungs and they were identified as females. The worms were cylindrical and annulated with a centrally located mouth surrounded by four hooks. The worms were rounded in cross section with tapering ends. Eggs were composed of two shell membranes. This is one of the new reports from rat snakes in Southern India.

Blastocystis sp. from food animals in India.

Blastocystis, a zoonotic protozoan found in the intestinal tracts of a wide range of animals, has not been reported from non-human hosts from India so far. Organisms indistinguishable from Blastocystis sp. were identified in the Giemsa stained intestinal scrapings collected from carcasses of piglet and poultry that were brought for necropsy to the Central University Laboratory, Chennai. The 'central vacuole forms' of the parasite, with number of nuclei ranging from 1 to 12 were identified. The intensity of infection was low, with less than one organism per oil immersion field, indicating that their presence was unconnected to the cause of death. Caecal scraping was found to be more ideal than duodenal scraping for the diagnosis of Blastocystis, and can be a potential specimen for definitive diagnosis. Identical organisms were also detected in the dung samples of a buffalo calf which showed clinical signs of diarrhoea The presence of Blastocystis in food animals acquires public health significance, as many subtypes of the parasite from poultry and pigs are transmissible to humans.

Multiplex PCR assay using SCAR primers to detect Eimeria spp. in chicken.

About 11 faecal samples from various regions of Tamil Nadu and Andhra Pradesh containing mixed spectrum of Eimeria species detected by morphometry viz. 15.4 × 11.2, 28.5 × 20.3, 31.1 × 18.5, 13.2 × 12.4, 20.8 × 17.5 and 22 × 18 µm for E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix, E. praecox and E. tenella respectively were taken for the study. The oocysts were concentrated and purified using faecal harvest protocol. The genomic DNA is extracted as per kit protocol. The amplicons of sizes 539 bp (E. tenella) and 460 bp (E. mitis) obtained could be visualised in a single lane for the multiplex PCR assay using sequence characterized amplified region primers.

Fusant Trichoderma HF9 with enhanced extracellular chitinase and protein content.

Strain improvement was carried out to obtain higher chitinase and protein by inter-specific protoplast fusion between Trichoderma harzianum and Trichoderma viride. Fusant HF9 and parental strains of Trichoderma were compared for chitinase and protein production. 1% of glucose, sucrose and fungal cell wall (Rhizoctonia solani), were used as carbon source for cultivation of Trichoderma and fungal cell wall was the best to induce chitinase and protein. Usage of 0.5% colloidal chitin for the fungal growth under aerated conditions at pH 6.5 and 28 degrees C led to higher chitinase and protein production. In these conditions fusant Trichoderma HF9 in comparison with parent strains had 3-, 2.5- and 1.5-fold increase of total chitinase, specific chitinase and protein, respectively. SDS-PAGE analysis revealed that it had 9 major protein bands with up-regulation compared to parent strains. Amino acid analysis showed that protein of culture filtrate of T. harzianum, T. viride and fusant Trichoderma HF9 had 8, 6 and 10 amino acids, respectively. The results obtained suggested that fusant HF9 could be an integration of T. harzianum and T. viride through protoplast fusion.

Molecular prevalence and preponderance of Eimeria spp. among chickens in Tamil Nadu, India.

Coccidosis is one of the most commonly prevalent and economically important parasitic diseases of poultry worldwide. Chicken coccidia are protozoan parasites of the genus Eimeria. This study aimed at analysing the molecular prevalence of seven species of Eimeria infecting chickens in Tamil Nadu, India. Tissue samples (caecum, rectum and upper and mid intestines) collected from chickens exhibiting symptoms of coccidiosis were used for DNA extraction, followed by amplification of the internal transcribed spacer (ITS) region of Eimeria genome with genus-specific primers and speciation in nested polymerase chain reaction (PCR) with species-specific primers. Of 43 tissue samples examined, 25 were positive in ITS PCR and all the seven species could be identified. However, the prevalence of each species varied. In broilers, Eimeria necatrix was present in all infected chickens with Eimeria brunetti, Eimeria tenella, Eimeria maxima and Eimeria acervulina present in more than 50% of infected chickens, while Eimeria praecox and Eimeria mitis were only present in 11% to 16%. Although only 7 samples were positive among layers, the prevalence was largely similar, but with a higher prevalence of E. praecox and E. mitis and a lower prevalence of E. tenella. Multiple infections were most common, with 2-6 Eimeria species infecting the same chickens. In order to estimate the preponderance of each infecting species of Eimeria, a random cloning technique was adopted. The genus-specific ITS PCR product was cloned in a TA vector and ten clones were randomly picked and used as template for amplification of all the seven genera of Eimeria. If the specific species of Eimeria is preponderant, then the frequency of the clones showing that species-specific PCR amplification would be higher. Using this method, the most preponderant species present in the rectum, mid and upper intestines of layers was assessed to be E. acervulina, E. brunetti and E. necatrix. E. acervulina was present in 60-90%, E. necatrix in 10-30% and E. brunetti in 10-20% of the clones screened, indicating that these species could be the most preponderant Eimeria species. Intervention strategies should aim at these species. This new method of estimating preponderance of infecting Eimeria species could be used to assess the relative importance of each species at the farm or region level instead of relying only on prevalence estimates.

In vivo immunomodulatory effects of ixodid ticks on ovine circulating T- and B-lymphocytes.

Selected aspects of the ovine immune system were examined during the course of repeated infestations with the ixodid ticks, Haemaphysalis bispinosa and Hyalomma anatolicum anatolicum that naturally infest sheep, either individually or together. By the use of flow cytometry it was shown that total T-lymphocyte numbers were significantly reduced from the sixth through the ninth days of all infestations. Gamma/delta (gammadelta+) and CD8+ T-lymphocytes were significantly depleted during tick feeding in all infested groups. CD4+ T-lymphocyte levels were significantly increased during secondary H. bispinosa and mixed species infestations. Hyalomma anatolicum anatolicum caused a significant increase in circulating B-lymphocytes over several days in both initial and secondary infestations. All infested sheep had increased CD4/CD8 and decreased T/B lymphocyte ratios during exposure to both ticks. Bromodeoxyuridine (BrdU) ELISA was used to measure in vitro proliferation of peripheral blood mononuclear cells stimulated with the T-lymphocyte mitogen Concanavalin A (Con A) after their collection from infested sheep. Significant suppression of in vitro proliferation occurred during first and secondary infestations with H. bispinosa, H. a. anatolicum and with both tick species together, beginning on the sixth day of infestation in all cases. These important tick species of sheep significantly modulate the numbers of immune effector cells and proliferation of T-lymphocytes derived from infested animals.