It is a challenging task to control internal parasites in grazing livestock even by applying multi label and multi directional approach. It is impossible to draw general recommendations to control parasitic diseases due to varied geo-climatic conditions and methods adopted for rearing the livestock in the country like India. In view of increasing incidence of anti-parasitic drug resistance in animals, there is an urgent need to design sustainable parasite control strategy which must include on the host as well as off the host control measures to harvest the maximum productivity from the animal for an indefinite period.

Aim:An environment compatible technique to stain Platyhelminthes, Fasciola gigantica, Gastrothylax crumenifer, Taenia solium, and Moniezia expansa using aqueous and alcoholic extract of sugar beet (Beta vulgaris), China rose (Hibiscus rosa-sinensis), and red rose (Rosa hybrida) were described to minimized the deleterious effects of the synthetic dyes.Materials and Methods:Aqueous/ethanolic extracts of roses were extracted from the flowers while red beet was extracted from the roots.Results:Stained helminthes acquired a comparable level of pigmentation with the distinction of their internal structure in these natural dyes. The flukes (liver and rumen) internal structure, oral and ventral/posterior sucker, cirrus sac, gravid uterus, testes, ovary, and vitallaria were appeared pink color in aqueous and alcoholic extract of either China or red rose and yellow to brown color in sugar beet stain. The interior of the proglottid of T. solium and M. expansa took yellow to brown color with good contrast in sugar beet stain and of pink to pink-red in China and red rose stain.Conclusion:The extract of roses (red rose followed by China rose) followed by red beet possess the potential to replace the conventional stains in the taxonomic study of Platyhelminthes parasites.

In an attempt to develop a suitable serological test for early detection of Fasciola gigantica infection in buffaloes, a group of proteins were isolated from the somatic antigen of the parasite by immunoaffinity chromatography. The process of isolation of the proteins has been standardized and significant level of repeatability was achieved. To test the diagnostic potentiality of the antigens, two serological tests, viz., enzyme-linked immunosorbent assay (ELISA) and dot enzyme-linked immunosorbent assay, were standardized using the sera from experimentally noninfected (group A) and infected (group B) animals. Further, the sensitivity and the specificity of the tests were evaluated employing the field sera from animals of different parasitic load viz., F. gigantica positive (group C), F. gigantica and Gastrothylax crumenifer positive (group D), F. gigantica and Gigantocotyle explanatum positive (group E), a group of sera without F. gigantica but other trematode infection (group F), only G. crumenifer positive (group G), only G. explanatum positive (group H), G. crumenifer and G. explanatum positive (group I), and PM negative (group J) collected from slaughterhouses of Bareilly (Uttar Pradesh, India) and Patna (Bihar, India). In plate ELISA, the sensitivity of the antigen and the test was 75.75% while the specificity was 97%, 95%, and 98%, respectively, against G. crumenifer, G. explanatum, and mixed infection of G. crumenifer and G. explanatum, respectively. In the case of dot ELISA the sensitivity was 86.5% and specificity was 92.3%, 94.7%, and 90%, respectively, against G. crumenifer, G. explanatum, and mixed infection of G. crumenifer and G. explanatum, respectively. The potentiality of the antigen in the diagnosis of field infection is discussed.

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Th1 and Th2 cytokine gene expression in buffalo calves during primary infection with Fasciola gigantica as well as in response to immunization with the parasite recombinant fatty acid binding protein (rFABP) and recombinant glutathione S-transferase (rGST) proteins was measured at 14th week of infection by real-time PCR with the double-stranded DNA-binding dye SYBR Green. Experimental animals were randomly distributed into FABP, GST, cocktail, challenge and healthy groups. Animals in groups FABP and GST were immunized with 400 μg rFABP and rGST, respectively, and cocktail group with a mixture of 400 μg each of rFABP and rGST in the neck and thigh muscles. All animals received three immunizations at 3-week interval. Calves were challenged per os with 400 viable metacercariae along with the unimmunized challenge control group 1 month after the last immunization. Expression of various cytokines in response to the immunization and parasite primary infection was measured by real-time PCR. Expression of IL-2 (4.5-fold) and IFN-γ (3.2-fold), followed by IL-6 (1.7-fold) and IL-4 (1.6-fold), with downregulation of TNF-α and IL-10 was observed in response to F. gigantica infection in these animals. However, there was a sharp increase in the expression of the IL-4 (211.93 and 111.81-fold) and IL-6 mRNA (219.22 and 48.29-fold) to GST and FABP immunizations, respectively. A downregulation of the IL-1α, a Th1 cytokine in response to FABP and GST immunization in these calves, was also observed. Overall, a mixed type of Th1 and Th2 cytokine environment was evoked to chronic F. gigantica infection and immunization with the above two recombinant proteins in buffaloes.

Fasciola gigantica, causative agent of tropical fasciolosis, inflicts substantial economic losses on the livestock industry, affecting severely buffalo productivity in the tropical countries. Very few vaccination trials with different target antigens against F. gigantica infection have been conducted in this host. Present study describes a vaccination trial in buffaloes with F. gigantica recombinant glutathione S-transferase and fatty acid binding protein. The two recombinant proteins were expressed in Escherichia coli and evaluated for their immunoprophylactic potential in buffalo calves, using montanide 70 M-VG, a mineral oil-based adjuvant, for delivering the antigens. Buffalo calves were distributed in three groups, with group I, II and III calves immunized with recombinant glutathione S-transferase, fatty acid binding protein and a cocktail of these two antigens, respectively. Immunization of the calves evoked a mixed IgG1 and IgG2 antibody response. Present vaccination trial in these animals achieved a maximum protection level of 35%, when the two antigens were used in combination. Eosinophils were measured in both immunized and non-immunized challenge control animals, which showed a steady increase in their count in response to immunization with both the antigens and infection with F. gigantica, respectively.

Out of 428 faecal 178 (41.59%) animals were found positive for Cryptosporidium parvum oocysts in the bovine faeces in modified Ziehl-Neelsen (mZN) staining and/ or commercial plate and dipstick-enzyme linked immunosorbent assay (ELISA) kits (Cypress diagnostics, Langdorp, Belgium) with statistically non-significant difference in the occurrence rate in cattle and buffalo calves. Seasonal prevalence difference was statistically significant in cattle calves while non-significant in buffalo calves, respectively. The highest overall prevalence was recorded during rainy season (45.15%) and almost same per cent during winter and summer. Prevalence of 67.26, 37.11, 30 and 17.65% was recorded in calves aged below 1 month, 1-3 months, 4-8 months and 9-12 months, respectively (p<0.05). There was no significant difference in the age group prevalence between cattle and buffalo calves. Both sexes of bovines are equally susceptible to the cryptosporidiosis (p>0.05). The prevalence of cryptosporidiosis was 59.54 and 29.41% for diarrhoeic and non-diarrhoeic samples, respectively (p<0.05). Both diarrhoeic and non-diarrhoeic groups of calves aged between 1-30 days were equally susceptible to the infection of Cryptosporidium spp. Almost same per cent prevalence of the infection was observed in dairy calves reared in organized (40.76%) and unorganized farms (42.21%) and the difference was non-significant in both cattle and buffalo calves. Highest prevalence of cryptosporidiosis was found in HF cross calves. The tests failed to detect the oocysts in infected soil samples. There was highly significant difference in the prevalence of cryptosporidiosis estimated by plate/ dipstick-ELISA and mZN staining with highest sensitivity and specificity in plate-ELISA.

Aim:Preservation of macroparasites by infiltrating the polymer in the tissues can defy the inherited shortcoming of classical wet preservation method.Materials and Methods:Preservation was done by infiltrating the melamine alone or with xylene (MX)/chloroform (MC)/turpentine oil (MT) in 1:1 and hardener (MH) in 9:1 ratio in the tissues of the gross specimen of the animal parasites.Results:The plastinated models withstand the process of microbial decomposition, and remain intact in the environmental conditions. The polymer mixture resists the entry of the water molecule, and model dried just after taking out it from the water tank. Overall, the plastinated parasites were dry, non-sticky, glossy, odorless, chemical free, and harmless, to some extent flexible, with detectable morphological structure, and retain their natural form but lost their natural color. Full marks were assigned to the degree of dryness, non-stickiness, and odorlessness to the model plastinated in different solutions on a five-point scale. For flexibility, the score was 1.2, 2.2, and 2.4 for the plastinated model in melamine/MH, MX/MC, and MT solutions, respectively. The average score of glossiness was 4.6 and 5 for the specimen plastinated in melamine/MH and MX/MC/MT solutions, respectively. The degree of dryness, glossiness, stickiness, and flexibility varies non-significantly, with the polymer mixtures used.Conclusion:The prepared model can be used to educate the students/general mass population.