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Control of Coccidiosis in Calves by Vaccination

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Control of Coccidiosis in Calves by Vaccination iolog ter y & c P a a B r f a o s i l Journal of Bacteriology and t o a l n o r g u y o J Parasitology Sultana, et al., J Bacteriol Parasitol 2014, 5:4 ISSN: 2155-9597 DOI: 10.4172/2155-9597.1000197 Research Article Open Access Control of Coccidiosis in Calves by Vaccination Razia Sultana1*, Azhar Maqbool2, Mansur-Ud-Din Ahmad2 , Aftab Ahmed Anjum2, Shabnum Ilyas Ch1 and Muhammad Sarfraz Ahmad1 1Department of Livestock and Dairy Development, Govt. of Punjab, Lahore, Pakistan 2Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan *Corresponding Author: Razia Sultana, Department of Livestock and Dairy Development, Govt. of Punjab, Lahore, Pakistan, Tel: +923014441753; E-mail: [email protected] Rec date: June 19, 2014; Acc date: Jul 29, 2014; Pub date: Aug 05, 2014 Copyright: © 2014 Razia S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract The immunizing effect of inactivated sporulated oocyst and inactivated sonicated vaccines against bovine coccidiosis was observed in calves. Indirect haemagglutination (IHA) test was developed for detecting antibodies to coccidian. Serum antibody levels in calves were measured against soluble oocyst (sporulated) antigen. IHA antibody titer was significantly higher (P<0.05) in calves vaccinated with inactivated sonicated vaccines as compared to the calves vaccinated with inactivated sporulated vaccines. Results of the challenge experiments indicated that the inactivated sonicated vaccine gave protection to the challenge calves as immune calves contained high level of antibodies that resisted heavy dose of challenge. Disease was observed in control group (non-vaccinated) calves post challenge whereas vaccinated remained healthy. Then the oocyst count per gram of faeces was significantly higher (P<0.05) in control group (unvaccinated) as compared with the vaccinated groups. Keywords: Calves; Coccidiosis; Vaccination; Sporulated oocyst retarded growth. Overcrowding, poor sanitation and poor nutrition are contributing factors for coccidiosis [6]. Out breaks of coccidiosis in Introduction calves and feeder cattle may be handled by mass medication using sulfonamides, amprolium or monensin added in feed or water [4]. Coccidiosis, a protozoal disease of many mammalian, and all Although it is possible to immunize cattle artificially however, the domestic livestock species, is caused by infection with species of the development of commercial vaccines appears difficult and vaccination genera Eimeria or Isospora. Clinically it is characterized by enteritis is not available as an alternative for treatment currently and in the although subclinical infections are frequent. The disease is relatively foreseeable future [7]. At present coccidiosis in cattle has neither given common in sheep, goats and cattle, rare in horses [1]. The overall importance in most of developing countries, Keeping in view the prevalence of coccidia was 47.1%, with the highest prevalence in <4 importance of disease in calves, this research project was design to months old calves (51.8%) and the lowest in >12 months old cattle evaluate an experimental vaccine as a potential candidate of control (27.0%). The number of oocysts per gram of feces was significantly program. higher in young calves than in weaners and adults [2]. Eleven species of Eimeria infecting the host, i.e., Eimeria alabamensis, Eimeria Materials and Methods auburnensis, Eimeria bovis, Eimeria brasiliensis, Eimeria bukidnonensis, Eimeria canadensis, Eimeria cylindrica, Eimeria ellipsoidalis, Eimeria pellita, Eimeria subspherica, and Eimeria zuernii. Vaccine Preparation The typical host for all isolated species is cattle. The most prevalent Culturing of coccidial oocyst: Oocysts of Eimeria bovis, recovered species was E. bovis (29.7%), while E. brasiliensis was the rarest (0.5%). from the naturally infected calves were sporulated [8]. The fecal Age-related analysis revealed a higher percentage of Eimeria spp. samples containing oocysts were placed in Petri-dishes containing 2.5 positive samples and higher OPG values in bison under 1 year old as percent potassium dichromate (K2Cr2O7) solution. These Petri dishes compared to older individuals (93.3% and 50-4050; 37.3% and 50-550, were then incubated at 30°C and sporulation time was recorded. respectively). Additionally, greater eimerian species diversity was Potassium dichromate solution was used as preservative. The present among calves in comparison with older bison [3]. Coccidiosis examination for recording the sporulation time was initiated at least produces bloody scours, bloody diarrhea, loss of weight and death. 12 hours post incubation of the Petri dishes and then continued every Most cases of Coccidiosis occur during winter but outbreaks may day until all the oocysts sporulated. occur sporadically throughout the year. Bovine Coccidiosis occurs most frequently in calves between six-to-twelve months of age [4]. The Counting of sporulated oocysts: Oocysts in 1 ml of faecal most serious losses are seen in dairy herds where large numbers of suspension were counted following the McMaster’s counting calves are kept along with older cattle carriers [5]. Calves become technique [9]. infected by ingesting sporulated oocysts along with their food and Harvesting of oocysts: The faecal material incubated in Petri-dish water. The severity of disease in calves depends upon the number of was centrifuged at 2000 rpm for 5 minutes and supernatant was sporulated oocysts they ingest. No symptoms are evident if few oocysts discarded. The pelleted oocysts were re-suspended in PBS and re- are ingested. Disease is severe if large numbers of oocysts are ingested. centrifuged. The procedure was repeated to give four washings. After In some cases death may occur. Sub clinical infection may cause J Bacteriol Parasitol Volume 5 • Issue 4 • 1000197 ISSN:2155-9597 JBP, an open access journal Citation: Sultana R, Maqbool A, Ahmad M, Anjum AA, Ilyas ChS (2014) Control of Coccidiosis in Calves by Vaccination. J Bacteriol Parasitol 5: 197. doi:10.4172/2155-9597.1000197 Page 2 of 5 last washing the pelleted oocysts were re-suspended at 50,000 oocysts Sheep erythrocytes were sensitized with glutaraldehyde and per ml [6]. sonicated antigen followed the method of Tokuda and Warrington [11] with modifications. The sensitized erythrocytes were finally Preparation of sonicated antigen resuspended in Phosphate Buffered Saline (PBS) to make 1.5% suspension. A two fold serial dilution of the serum samples was made The final concentration of 50,000 oocysts per ml was subjected to with PBS. Equal volume (0.05 ml) of sensitized erythrocytes (1.5%) ultra-sonication (8] at 60 htz for 5 shots of 1 minute each with an was added in each well of the microtitre plates. The plates were tapped interval of 30 seconds by placing the material on ice. Sonicated oocysts to ensure even mixing of erythrocytes and incubated at 37°C for 90 suspension was centrifuged at 2000 rpm for five minutes, supernatants minutes. The degree of haemagglutination in each well was recorded and sediments were collected separately for preparation of vaccines. in comparison with control. The highest dilution producing HA was recorded by observation suspended aggregates as positive and button Inactivation of oocysts formation as negative. Following vaccines were prepared by inactivating oocyst suspension by incubating with 0.3% formalin for 48 hours [10]. Biological Challenge Supernatant from sonicated sporulated oocyst (50000 sporulated On day 15 post-vaccination, all the calves in group I, II, III and IV oocysts/ml) were challenged by 20,000 sporulated oocysts of species Eimeria bovis. The faecal samples were collected weekly upto day 42 post- Sediment from sonicated sporulated oocyst (50000 sporulated vaccination. Numbers of oocysts per gram of faeces were calculated oocysts/ml) from each challenged group weekly till day 42. Clinical Sign were also Un-sonicated sporulated oocyst (50000 sporulated oocysts/ml) recorded. Safety Test Results and Discussion All the prepared vaccines were cultured to check the presence of any live oocyst Method of cultivation [8]. These vaccines were also Vaccine checked for the presence of any bacterial contamination by inoculating Safety Test: All the vaccines were checked for presence of any live the material of vaccines on blood agar and vaccines found unreliable parasites and were found free. Vaccines were also cultured on blood were discarded. agar and were free of any bacterial contamination. Immune response of experimental vaccine Immune Response of Experimental Vaccines The immune response of experimentally prepared vaccine was The geometric mean antibody titers of vaccinated and non- studied in calves under one month of age. A group of 20 cattle calves vaccinated groups by Indirect Haemagglutination test (IHA) are under one month of age were purchased from the local market and presented in Table 1. The highest Antibody titer (512) was recorded in reared under standard manage mental conditions. Calves on day 6 group (A and B) on day 35. Antibody titer (362) on day 35 in groups C were equally divided into four groups. The detail for each group is as was lower as compared to group A and B. under: Indirect Haemagglutination test (IHA): Antibody titer (geometric Group-I: Vaccine A was given at the dose rate
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