Diagnosis of cystic echinococcosis in buffaloes by native 8 kDa antigen using latex agglutination test (LAT).

The dog tapeworm Echinococcusgranulosus is the causative agent of cystic hydatid disease in domestic/wild herbivores animals and man. Accurate immunodiagnosis of the infection requires highly specific and sensitive antigens. The aim of this study was to develop and evaluate immunoassays with principles of precipitation, agglutination for the identification of buffaloes infected with hydatid cyst which would allow the monitoring of animals from endemic areas and identifying infected animals prior to slaughter. The immunoassays were developed and validated using hydatid specific, non-cross reactive low molecular weight 8 kDa hydatid cyst fluid protein. Sera used for the assay validations were obtained from 200 buffaloes infected naturally with hydatid cyst and 200 non-infected buffaloes. The diagnostic sensitivity with latex agglutination test was 98.67 %. It should be useful for the conformation of hydatid cyst infected individual sheep.

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.

Assessment of cell mediated immune response in rabbits immunized with affinity purified 35 kDa midgut antigen of Rhipicephalus haemaphysaloides ticks.

Cell mediated immune response to immunoaffinity chromatography purified midgut antigen of Rhipicephalus haemaphysaloides ticks in rabbits was studied by using lymphocyte transformation test. This test was carried out by using 5-bromo-2'-deoxy-uridine kit method. The blastogenic response of peripheral blood lymphocytes of normal rabbit to different concentrations of antigen and mitogen (Con A) showed that 2 µg of antigen and 2 µg of mitogen gave maximum stimulation index. The antigen specific responsiveness of immunized rabbits with affinity purified 35 kDa midgut antigen was highly significant (P ≤ 0.01) compared to mitogen. The maximum lymphocyte stimulation index (LSI) of 2.47 was observed on 49 day post immunization in immunized group. The lymphocytes separated from control animal cultured in RPMI1640 medium with 2 µg of antigen and 2 µg of mitogen (Con A) were never stimulated and their LSI values were below 2.0.

Sparganosis in Russell's viper snake: a case report.

The occurrence of the plerocercoid larva, Sparganum of Sparganum spp. in two Russell's viper snakes maintained in Chennai snake park trust, Chennai is reported for the first time from Southern India. The cestode larvae were found in the sub cutis and were flat, solid, wrinkled, ribbon like creamy white in colour with peudosegmentation with anterior end possessing bothria. The cuticles, subcuticular cells and parenchymatous tissue stained by acetic alum carmine further confirmed the cestode larvae.

Partial sequencing of Bm86 gene for studying the phylogeny of an Indian isolate of Rhipicephalus (Boophilus) microplus tick.

Tick gut glycoprotein, designated as Bm86, found on the luminal surface of the plasma membrane of gut epithelial cells of Boophilus microplus, which is a concealed antigen, has been used as vaccine candidate molecule for immunization against ticks. To better understand the molecular diversity of Bm86 gene in ticks, a portion of the cDNA was sequenced from an Indian isolate of B. microplus. Comparison of nucleotide sequence revealed that Indian isolate had 97 % homology (18 polymorphisms) with that of the Australian isolate and 96 % homology (20 polymorphisms) with that of the Cuban vaccine strain. Further, the Indian isolate differed from the Cuban vaccine isolate at 7 amino acid loci, including 5 substitutions (at residues 88, 94, 175, 176 and 177) and 2 deletions (at 183 and 184). However, protein prediction studies did not show any difference in the putative antigenic epitopes of the protein expressed.

Kalicephalus sp. in a captive Russell's viper: a case report.

The occurrence of Kalicephalus sp. of hookworms in a Russell's viper snake maintained in Chennai snake park trust, Chennai is reported. These worms were found in the stomach and intestine. They were cylindrical, thread like and had an elongate body. Anterior end was obliquely truncated with bivalvular buccal capsule. Short, thick and muscularised oesophagus ending in a rounded bulb was observed. In males, a well developed trilobed bursa was observed and the spicules were short and equal. Typical strongyle type eggs were found in the uterus of female worms as well as in the intestinal contents.

Low hemoglobin density as a measure of iron status.

Iron deficiency anemia is the commonest cause of anemia in the developing countries. Iron status is the result of the balance between the rate of erythropoiesis and the amount of iron stored in the body. Various biochemical parameters have been used to assess iron status such as iron levels, transferrin, transferrin saturation and ferritin, and all of them may be influenced by acute phase response and are also expensive tests 1-4. In our situation where patients cannot afford exhaustive tests to document iron deficiency we utilized the LHD values as a predictor of iron status based on the formula provided by Urrechaga 5.

Factors influencing on prevalence of Cryptosporidium infection in south Indian dairy calves.

The objectives of the present study were to report the influence of factors like age, sex, breed, dung consistency and rearing system on prevalence of Cryptosporidium spp. in south Indian cattle. Two-step nested PCR was employed for detection of Cryptosporidium infection in dairy calves of south Indian states viz., Andhra Pradesh, Karnataka, Kerala, Tamil Nadu and union territory i.e., Puducherry. A total of 459 dung samples from the calves were subjected to nested PCR, 182 were found positive with prevalence percent of 39.65. Age wise comparison showed a high prevalence of Cryptosporidium in the age group of one month old calves. This concludes that the cryptosporidiosis is highly age dependent with young calves showed the highest prevalence. Depending on the group had consistency of dung, the highest prevalence of Cryptosporidium was observed in semi-solid dung, followed by formed and the diarrhoeic group animals. Female calves showed slightly higher prevalence rate than male animals. Cow calves had an overall prevalence percent of 40.75 and the infection rate in buffalo calves was 36.28 %. In relation to rearing system, individual animals had 42.18 % and farm animals showed 38.46 % of Cryptosporidium infection. In conclusion, the prevalence of Cryptosporidium in dairy calves should be correlated with the factors like age, sex, breed, dung consistency and rearing system of the animal to arrive at a reliable epidemiological data on bovine cryptosporidiosis.

Molecular prevalence of Cryptosporidium spp. in dairy calves in Southern states of India.

Dung samples were collected from dairy calves of south Indian states viz., Andhra Pradesh, Karnataka, Kerala, Tamil Nadu and union territory, Puducherry and are subjected to nested polymerase chain reaction (PCR) targeting 18S rRNA gene for detection of Cryptosporidium infection. Of the 459 dung samples screened 182 were found positive with a prevalence of 39.65%. Highest prevalence of Cryptosporidium was observed in Puducherry (86.67%) and lowest in Kerala (17.65%). Genotyping by PCR-restriction fragment length polymorphism (RFLP) and sequence analysis revealed the presence of all the four major Cryptosporidium species of cattle viz., Cryptosporidium andersoni, Cryptosporidium ryanae, Cryptosporidium parvum and Cryptosporidium bovis. C. andersoni was widely distributed in calves of Tamil Nadu, Karnataka and Puducherry whereas in Andhra Pradesh C. ryanae was the major species. Of the 64 samples subjected to PCR-RFLP, 39 (60.94%) could be classified as C. andersoni, 18 (28.13%) as C. ryanae, 4 (6.25%) as C. parvum and 3 (4.69%) were confirmed as C. bovis. The results were also confirmed by sequencing of 19 Cryptosporidium DNA samples.

Occurrence of Laemobothrion maximum in Greater Coucal.

External examination of free range abandoned dead Greater Coucal in a veterinary dispensary, Vellore, Tamil Nadu, India revealed presence of a large sized louse in the feathers and was identified as Laemobothrion maximum on the basis of morphology and size of the louse.

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.

Seasonal activity of ticks on small ruminants in Tamil Nadu State, India.

The seasonal activity of ticks on sheep and goat populations was examined from December 2000 to November 2001 in the state of Tamil Nadu, India. This longitudinal study spread over four seasons and seven agroclimatic zones took into consideration the spectrum of species involved, the levels of infestations, the seasonal epidemiology of ticks and the associated management practices. The most common species of tick spread throughout the state was Haemaphysalis bispinosa followed by Rhipicephalus haemaphysaloides, Hyalomma marginatum isaaci and Hyalomma anatolicum anatolicum with mixed infestations being the rule. Although the infestations were found throughout the year, they were greater during the rainy season and rainfall seemed to be the most important climatic factor affecting seasonal variation. In general, most of the small-ruminant populations carried moderate tick burdens with mixed infestations and this varied with the management practices.

Phylogenetic relationships of Hepatozoon (Apicomplexa: Adeleorina) based on molecular, morphologic, and life-cycle characters.

To evaluate higher-level affinities of Hepatozoon species within Apicomplexa, we sequenced the 18S rRNA gene from 2 parasites (Hepatozoon americanum and Hepatozoon canis) of dogs and 1 (Hepatozoon catesbianae) of bullfrogs. Sequences from other apicomplexans among the Sarcocystiidae, Eimeriidae, Theileriidae, Plasmodiidae, Cryptosporiidae, and Babesiidae, a Perkinsus species and 2 dinoflagellates were obtained from GenBank. Phylogenetic analysis indicated that Plasmodium, Cryptosporidium, and Hepatozoon form a monophyletic group distinct from representatives of other apicomplexan families. Although equivocal, our analysis indicated that Plasmodium and Cryptosporidium are sister taxa and that Hepatozoon is basal to them. To evaluate phylogenetic affinities among H. americanum, H. canis, and other species of Hepatozoon, we examined 18 morphologic and life-cycle features of 13 species currently assigned to Hepatozoon. This analysis indicates paraphyly of Hepatozoon (as currently arranged) because Hepatozoon lygosomarum was found most closely related to Hemolivia mauritanicum. These results, combined with results of previous studies, support elevating Hepatozoon to familial level (Hepatozoidae) as originally suggested by Wenyon in 1926. Both DNA sequence data and morphologic and life-cycle characters support a sister-group relationship between H. americanum and H. canis.