Value of Serum and Milk ELISAs in Diagnosis of Fasciolosis in Lactating Buffaloes Using Fasciola gigantica Partially Purified Somatic Antigen (original) (raw)
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Detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay
Parasitology Research, 2008
The process of isolation of the 27-kDa glycoprotein from the somatic antigen of Fasciola gigantica was standardized and the diagnostic potentiality was evaluated for the detection of bubaline fasciolosis by indirect enzymelinked immunosorbent assay. Initially, the test was standardized using the sera from experimentally noninfected(n=20) and infected (n=5)animals. Further, the sensitivity and the specificity of the test were evaluated through the sera of buffaloes with different natural infections, i.e., F. gigantica (n=8 animals), F. gigantica and Gastrothylax crumenifer(n= 15), F. gigantica and Gigantocotyle explanatum (n=6), trematode infections other than F. gigantica (n=9), only G. crumenifer (n=36), only G. explanatum (n=18), G. crumenifer and G. explanatum positive (n=39), and PM negative (n=102). All animals came from the slaughterhouses of Bareilly (Uttar Pradesh, India) and Patna (Bihar, India). The level of sensitivity observed in the present study was 81.0%, while 97-98% specificity against G. crumenifer, G. explanatum, or a mixed infection with both parasites was noted. The study showed F. gigantica prevalence rate of 18-20% in the buffaloes of the study area. Enzyme-linked immunosorbent assay with a 27-kDa glycoprotein could be a feasible diagnostic method for the early detection of bovine fasciolosis.
The Enzyme-Linked Immunosorbent Assay (ELISA) was evaluated for the diagnosis of Fasciola gigantica infection in cattle and buffaloes. The excretory-secretory (E-S Ag) antigen of Fasciola gigantica adult flukes obtained after invitro incubation was used as an antigen. The test was conducted with 276 sera collected from cattle and buffaloes which included 22 sera each from naturally infected cattle and buffaloes (known positive serum) and with similar number of samples with healthy cattle and buffaloes (known negative serum). The positive results were observed in 18 and 19 of the sera from naturally infected cattle and buffaloes with sensitivity of 81.8% and 86.3% respectively. Out of 188 serum samples which were found negative on faecal examination 32 (34%) sera of cattle and 40 (42.5%) sera of buffaloes were found positive by ELISA respectively. The sensitivity of the test was found to be 91.6% and 95.6% in cattle and buffaloes respectively.
Parasitology Research, 2008
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.
Assiut Veterinary Medical Journal, 2014
The present study was carried out to investigate the prevalence of fascioliasis among 100 cattle and buffaloes of different age and sex in Assiut Governorate. Samples were examined by microscopic examination, Agar Gel Diffusion Test AGDT and ELISA techniques to detect Fasciola species eggs and antibodies, respectively. Prevalence of Fasciola was 8% by microscopic examination, 32% by AGDT and 60.86% by ELISA test. Cattle showed higher prevalence of infection 61.4% than buffaloes 29.4%. Female cattle and buffaloes were more susceptible to the infection 64.55% than male 23.8%. Fasciola was more frequently recorded in 2-4 years old cattle and buffaloes 76% followed by more than 4 years 46.7% then less than 2 years of age 20%. All positive results with fecal egg examination and AGDT were also positive with ELISA. Serological examination by AGDT and ELISA confirms the microscopical examination. The results of this study revealed that the agar gel diffusion test and ELISA could become a useful tools to diagnose fascioliasis in cattle and buffaloes.
Journal of Parasitic Diseases, 2015
This survey was done to investigate the efficacy of the in-house indirect ELISA (iELISA) and Dot-ELISA methods Prepared from excretion-secretory (ES Ag) and Crude (Cr Ag) antigens of Fasciola for sero-diagnosis of Fasciola gigantica in cattle. The liver specimens of slaughtered cattle were collected and their liver examined macroscopically and microscopically for infestation to fasciolosis. Sera from two groups of cattle, one infected with fasciolosis (n = 60) and the other non-infected with fasciolosis (n = 60), were used in the iELISA and Dot-ELISA test; grouping based on histopathology results. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of both Dot-ELISA and iELISA done with ES Ag were better than those of tests performed with Cr Ag. Interestingly, the reliability of two methods was very good similar to one another.
Comparative Susceptibility of Buffaloes and Cattle to Fasciola Spp. Infection in
Fasciolosis is one of the major veterinary problems in the world that reduces the animal production. In the present study, a total of 3600 faecal samples (1800 from each of buffaloes and cattle) was collected on a random basis from different areas of Bahawalpur between February and October, 2013. During sampling the animals body condition and their deworming history were recorded. An overall prevalence of fasciolosis in buffaloes and cattle amounted to 15.8% and 14.2%, respectively. Furthermore, in both buffaloes and cattle it was found that poor body conditions (43.9% and 48.8%, respectively) and no deworming (25.9% and 24.6%, respectively) favoured animal susceptibility to infection. Fasciola gigantica was the major infection in both buffaloes and cattle (11% and 10.1%, respectively) as compared to that of Fasciola hepatica (buffaloes 3.61% and cattle 2.9%
The present search was performed to detect a predominance of Fasciola hepatica in buffaloes in Wasit Province, to investigate the related risk factors and to evaluate the efficacy of coprological (floatation and sedimentation) tests in comparison with serological indirect ELISA that considered as a gold standard technique in diagnosis of a disease. Out of 46 randomly selected buffaloes examined by fecal and serum blood samples, 6.5, 10.9, and 37%; were positives by floatation, sedimentation and ELISA tests, respectively. With ELISA, the degree of infection in positives was 70.9% mild, 24.9% moderate and no one had been strong. In dependence on the risk factors, the results of sex, age, body condition and feces consistency groups showed a greatly significant differences (P>0.05) in infection rates that recorded by the diagnostic techniques that used in this study. In related to serum total bilirubin, there was no significant differences between jaundiced and non-jaundiced groups (P<0.05).
Designing and evaluation of Dot-ELISA for diagnosis of Fasciola infection in cattle
Veterinary research forum : an international quarterly journal, 2014
Fasciolosis is a disease caused by liver fluck of the genus of Fasciola. Diagnosis of fasciolosis has been challenging for a long period due to low sensitivity of the coprological diagnostic method. In this study, an in-house Dot-ELISA method; using excretion-secretory (ES Ag) and Crude (Cr Ag) antigens of Fasciola was described for diagnosis of fasciolosis in cattle. For this purpose, the sera specimens of slaughtered cattle were taken and examined for Fasciola infection. Sera from two groups of cattle, one infected with Fasciola (n = 60) and the other non-infected with Fasciola (n = 60), were used in the Dot- ELISA test. All sera were tested and evaluated. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of Dot- ELISA with ES Ag were better than those of Dot- ELISA with Cr Ag. In conclusion, excretory-secretory antigen dependent Dot-ELISA can be used as a reliable sero-diagnostic test for Fasciola infection in cattle.
Indian journal of experimental biology, 2005
A glycoprotein (27 kDa) was isolated from crude somatic antigen of Fasciola gigantica by two steps affinity chromatography and was used in early detection of experimental fasciolosis in cattle by indirect ELISA and in dot-ELISA formats. Although, anti-27 kDa antibodies could be detected after 3 weeks post infection (WPI) by dot - ELISA which was one week later than indirect ELISA. The test, dot-ELISA, was more convenient in field application. By the test (dot-ELISA) the infection could be equally detected in animals infected with 100, 200 and 300 metacercariae of F. gigantica with high sensitivity. Further, the antigen (27 kDa) was not found to react with goat sera infected with Paramphistomum epiclitum, which are giving strong reaction to homologous immature and mature fluke antigens of P. epiclitum.
A comparative study of three methods for detecting Fasciola infections in Nigerian cattle
Qualitative examination of Fasciola gigantica eggs in faeces and bile were compared with the detection of precipitating antibodies in sera by agar gel precipitation test (AGPT) in 1000 cattle slaughtered at the Bodija municipal abattoir in Ibadan, Nigeria. Faecal and bile examination methods detected (196) 33.5% and (389) 38.9% of the animals as positive for fasciolosis, while (474) 47.4% were positive by AGPT. Both direct bile examination and faecal egg detection methods have high specificity and positive predictive value (100%) when compared with AGPT. However, lower values for sensitivity and negative predictive value were observed for both faecal egg examination (66.5% and 67.9% respectively) and bile examination (81.0% and 78.9% respectively). Fecal and bile examination failed to detect 33.5% and 19.0% of the cases detected by AGPT. The results of this study revealed that the AGPT could become a better test for the herd diagnosis of bovine fasciolosis for veterinarians and othe...