Cryptosporidiosis in Calves, Lambs and Goat Kids in Bishoftu, Oromia Regional State, Ethiopia

African Journal of Basic & Applied Sciences 7 (5): 233-239, 2015 ISSN 2079-2034 IDOSI Publications, 2015 DOI: 10.5829/idosi.ajbas.2015.7.5.95160 Cryptosporidiosis in Calves, Lambs and Goat Kids in Bishoftu, Oromia Regional State, Ethiopia Dinka Ayana and Berhanu Alemu Addis Ababa University, College of Veterinary Medicine and Agriculture, Ethiopia Abstract: A cross sectional study was carried out from November 2014 to April 2015 to determine the prevalence of Cryptosporidiosis in calves, lambs and goat kids and to assess potential risk factors in Bishoftu, of Oromia regional state, Ethiopia. Hence, fecal samples from a total of 364 study animals (214 calves, 89 lambs and 61 kids) were examined with Sheather s flotation technique and Modified Ziehl-Neelsen Staining. Accordingly, the overall prevalence was found to be 14%. The prevalence of 13.6% (29/214) in calves, 16.9% (15/89) in lambs and 11.5% (7/61), in kids were recorded. There was a significant difference (P < 0.05) in the prevalence of cryptosporidiosis between diarrheic and non-diarrheic animals. There was no statistically significant variation (P>0.05) observed in the prevalence of Cryptosporidium infection between the study sites, animal species and among age groups. In conclusion, this study demonstrated the importance of Cryptosporidium in young ruminants with a higher prevalence among diarrheic animals than non diarrheic ones. Key words: Bishoftu Calves Cryptosporidiosis Kids Lambs Prevalence Risk Factors INTRODUCTION Cryptosporidiosis is an emerging protozoan disease, caused by Cryptosporidium species,that can cause Ethiopia possesses the largest livestock population gastrointestinal infection in a wide variety of mammals in Africa. Livestock is a significant contributor to including human, cattle, sheep, goat, pig and horses economic and social development in Ethiopia at worldwide [5]. The infection being encountered after household and national level. Livestock accounts for ingestion of the microscopic infective oocysts. Oocysts 15-17% of total GDP and 35-49% of agricultural GDP. are discharged in the feces of infected cattle and are of Livestock directly contributes to the livelihoods of more primary importance for the dispersal and survival of the than 70% of Ethiopians [1]. parasites [6]. Calves are primarily infected via the fecal- The country faces a range of opportunities and oral route and it takes less than 50 oocysts to infect a constraints improving the productivities of its livestock healthy calf [7]. Infection can rapidly spread from calf to population. The main constraints to increasing livestock calf when animals are commonly housed and overcrowded productivity and output are the lack of adequate supplies or from cow to calf via the udders when they are of good quality livestock food, high incidence of disease contaminated with infected calf feces in the lying area of and mortality rates and water shortage. Livestock disease the dams. These oocysts are resistant to the environment is the major constraints of productivity causing economic and remain infective for months in cold water or dump, losses to the peasant farmers and pastoralists in Ethiopia cool environment [5]. community to hundreds of millions of birr annually [2]. Clinical Cryptosporidiosis is frequently not Infectious diarrhea (scours) of neonatal animal is a diagnosed, yet it has been incriminated as an important common disease. The infectious agent that causes cause of diarrhea in neonates. Clinically, the disease is diarrhea can be a virus (Bovine viral diarrhea virus, characterized by anorexia and diarrhea, often intermittent, rotavirus, coronavirus), bacteria (E. coli, Salmonella, which may result in poor growth rate. The severity of enterotoxaemia), protozoa (coccidia, cryptosporidiosis, clinical disease may be associated with the animals Giardia) [3,4]. immune and nutritional status. It is also characterized by Corresponding Author: Dinka Ayana, Addis Ababa University, College of Veterinary Medicine and Agriculture, Ethiopia. 233

low morbidity which, however may become severe when incites caution in drinking unpasteurized milk. Milk can be associated with other pathogens [5], although calves 1-3 contaminated through mechanisms of poor udder hygiene weeks old seem to be most susceptible, Cryptosporidium and recent outbreaks of human cryptosporidiosis species has also been found in cattle over two years of associated with drinking unpasteurized milk have been age impairing rate of gain in feedlot cattle and milk reported [22, 23]. production in dairy cattle [8]. Furthermore, the large number of oocysts excreted Cryptosporidiosis occurs primarily in neonatal during infection helps to ensure a high level of calves, but also in lambs and kids [9].One major species, environmental contamination. Although not always C. parvum, infects both farm animals and humans [10]. confirmed, cattle facilities are frequently blamed when Cryptosporidiosis is a fairly prevalent disease in many Cryptosporidium is found in surface waters. Thus, cattle countries and the disease is one of the most economically living in close proximity to rivers should be considered important diseases especially in calves and kids in Turkey potential causes of waterborne contamination, as surface [11]. Calves at one to15 days are at the highest risk [12]. run-off does transport Cryptosporidium oocysts in soils The higher percentage of oocysts excreted is observed in to water sources [24]. 7-day-old calves. Animals of all ages can be infected, but For calves, lambs and kids Cryptosporidium has diarrhea occurs only in young animals [13]. In young become a concern not only because of the direct calves there is a significant relationship between season economic losses associated with the infection, but also and infection [12]. Cryptosporidium infections have now from a public health perspective because of the potential been reported in most domestic animals as well as in a for environmental contamination with Cryptosporidium large variety of wild and captive animals since its first oocysts. It is this zoonotic potential, the acquisition of identification by Tyzzer. Most infections have been C. parvum by humans from fecal contamination by described in mammals and are attributed to C. parvum animals, which may have the greatest impact on the dairy [14]. industry and food of animal origin. In1981, Young animals appear to be more susceptible to cryptosporidiosis was diagnosed in veterinary students infection and disease, while infections in adult animals are who cared for calves with cryptosporidiosis [25]. Recent often asymptomatic or do not occur. Similarly to human molecular studies of cryptosporidiosis in cattle have cryptosporidiosis, the common symptom in animals is shown that three species and one genotype of yellow watery diarrhea which leads to dehydration, weight Cryptosporidium are responsible for most cattle loss, fever and inappetence. Most (immunocompetent) infections (C. parvum, C. bovis and C.andersoni and animals recover within 1 2 weeks of infection with Cryptosporidium deer-like genotype) [26]. C. parvum is supportive fluid therapy [14]. Cryptosporidiosis in known to infect humans worldwide and is recognized as ruminant species is typically symptomatic in the young. the major zoonotic Cryptosporidium species, whereas Among cattle, calves are susceptible to infection shortly C. andersoni has been reported in humans only once [27]. after birth and remain so for several months [15]. Infection The most prevalent species were C. parvum in in dairy calves is most often detected (via fecal oocyst pre-weaned calves, Cryptosporidium bovis has been shedding) between 8 and 15 days of age, whereas described recently. C. bovis oocysts are morphologically infection in beef calves most often occurs between 1 and indistinguishable from C. parvum oocysts. 2 months of age [16,17]. Cryptosporidium bovis is a highly prevalent species that Infection in lambs and goat kids is more common in infects primarily post-weaned calves [28]. Among animals under 1 month old [18]. Infection can be spread parasitic diseases, coccidiosis and cryptosporidiosis are animal-to-animal by the fecal-oral route, usually when important protozoan parasites responsible for low animals are housed together in an overcrowded productivity and mortality in small ruminants. environment, but contamination of udders and water Cryptosporidiosis, caused by the genus supplies by feces is another common source of Cryptosporidium, is an important protozoan disease of transmission in livestock. Transmission of the bovine young animals and humans with a cosmopolitan genotype of C. parvum from calves to humans is distribution. Contrary to coccidiosis, the disease is not established [19, 20]. It is estimated that 50% or more of all host-specific and affects several species of animals [29]. dairy calves will shed detectable numbers of oocysts and These parasites do not require an external that the parasite is present on more than 90% of all US development stage, but are immediately infective when dairy farms [21]. This high prevalence of infected calves passed in the feces as thick-walled oocysts. Within hosts 234

such as calves, lambs and kids Cryptosporidium have The sample size was determined using the formula both sexual and asexual reproduction as well as a given by [32] with a 95% confidence interval since there complete life cycle within the calves, lambs and kids was a study taken by [33] and his prevalence was 17.6% through thin-walled oocysts. Through this mechanism, and at 5% absolute precision. Therefore, the total sample the population of the parasite in the individual host is size is 223. To increase, the precision level these number magnified [30]. was increased to 364 with the proportion of 214 (13.6%) A variety of methods is available for detection of calves, 89 (16.9%) lambs and 61 (11.5%) kids. Cryptosporidium species including microscopic, immunological and molecular techniques. Microscopic 2 2 n= 1.96 pexp(1- pexp)/d detection is based on finding the environmental and n= sample size, pexp= expected prevalence, d= precision chemical resistant oocysts in fecal samples [8]. Oocysts may be demonstrated using Ziehl-Nielsen stained fecal Sample Collection: Fresh fecal samples were randomly smears in which the sporozoites appear as bright red collected directly from the rectums of study animalsusing granules. In Ethiopia, there is limited information on the disposable gloves, placed in universal bottle and status of Cryptosporidiosis. Therefore, it is with this view transported in ice box to Veterinary parasitology and understanding that the present study is initiated with laboratory, CVMA, for processing on the same day. The the following specific objectives: samples were placed in suitable leak-proof plastic containers, tightly closed, labeled and transported to the To determine the prevalence of Cryptosporidiosis in laboratory for examination. The samples were first calves, lambs and kids in Bishoftu screened by Sheather s Floatation Technique [34] and To identify risk factors associated with the disease those found positives were confirmed by Modified Ziehl-Neelsen Stain [35]. MATERIALS AND METHODS Data Management and Analysis: The data were entered to Study Area: The study was conducted in Bishoftu, east excel spread sheet program to create a data base which shoa zone of Oromia region from November, 2014 to May, was transferred to SPSS 20.0 Version software program of 2015. Bishoftu is located 45 kms south east of Addis the computer before analysis.alltheparameters measured Ababa. The area is located at 9 N latitude and 40 E were analyzed by using SPSS 20.0 statistical software. longitudes at an altitude of 1850 meters above sea level in Significant level was determined at 95% confidence level central high land of Ethiopia. It has an annual rainfall of (P< 0.05). 866 mm of which 84% is in the long rainy season (June to September). The dry season extends from October to RESULTS February. The mean annual maximum and minimum temperatures are 26 C and 14 C respectively, with mean Among 364 animals examined using Ziehl-Neelsen relatively humidity of 61.3% [31]. staining technique, the overall prevalence of Study Population: Only young animals of age group Cryptosporidium in all species was found to be 14%. The below 12 month calves, lambs and kids were prevalence in calves was 13.6% (29/214) while the consideredforthis particular study. The association of the proportion in lambs and kids were 16.9% (15/89) and disease occurrence was determined in relation with 11.5% (7/61), respectively (Table 1). Significant difference different age category (5 days 6 months and >6 month-12 (P < 0.05) was observed in the prevalence of months), study sites, animal species (calves, lambs and Cryptosporidiosis between diahoaric and non-animals. kids) and fecal consistency (diarrheic and non diarrheic) However, there was no significant (P > 0.05) difference of calves, lambs and kids as well. observed in the prevalence of cryptosporidiosis among animal species, study sites (farms) and between the age Study Design and Sample Size Determination: A cross group of the study animals. sectional study design was used to determine the An examination of the consistency of the fecal prevalence of Cryptosporidium in calves lambs and kids. samples showed that 26.9% (98/364) of samples were Systematic random sampling technique was applied to diarrheic. Of these, 41.8% (41/98) tested positive for select each study animal. Cryptosporidium oocysts. The remaining 73.1% (266/364) 235

Table 1: The prevalence of Cryptosporidiosis by animal species Animal species Number examined Number positive % Chi-square P-value Goat Kids 61 7 11.5 0.960 0.619 Lambs 89 15 16.9 Calves 214 29 13.6 Total 364 51 14 Table 2: The prevalence rate of Cryptosporidiosis by farms Sites (farms) Number examined Number positive % Chi-square P-value Ethiopian milk and meat institute dairy farm 24 4 16.7 1.140 0.980 Ethiopian Agriculture Research center dairy farm 28 3 10.7 Genesis dairy farm 29 3 10.3 ALFA fooder and dairy farm 72 9 12.5 Tseday small holder dairy farm 15 2 13.3 Fair field integrated small holder farm 47 7 14.9 Extensive farm 149 23 15.4 Total 364 51 14 Table 3: Prevalence of Cryptosporidiosis by age group and fecal consistency Variables Number examined Number positive % 2 X P-value Age 5 days- 6 months 327 44 13.5 0.823 0.247 >6 months-12 months 37 7 18.9 Fecal consistency Diarrheic 98 41 41.8 86.184 0.000 Non-diarrheic 266 10 3.8 were non diarrheic, with 3.8% (10/266) of these testing prevalence in lambs and kids were (16.9%) and (11.5%) positive (Table 3). There was statically significant respectively. There was statically significant difference (P difference (P < 0.05) between the numbers of diarrheic <0.05) between diarrheic (41.8%) and non diarrheic samples with Cryptosporidium oocysts compared to the animals (3.8%). This was in agreement with the results of non diarrheic samples. The prevalence of previous studies showing that calves infected by entero Cryptosporidium significantly (p < 0.05) varied with fecal pathogens can be subclinically infected [39]. consistency of study animals being ten times higher In Europe, Cryptosporidium oocysts were detected likelihood of occurrence in diarrheic compared to its in 42.1% of the examined lambs and 31.8% goat kids in chance of occurrence in non diarrheic (OR = 76.488). Serbia [40]. The study conducted by Dinka Ayana et al. The prevalence rate of Cryptosporidium according to age [41] on Eimeria and Cryptosporidium infections in sheep groups of 5 days- 6 month and >6 month- 12 month were and goats at ELFORA export abattoir, Central Ethiopia 13.5% (44/327) and 18.9% (7/37) respectively. There was exceptionally reported zero prevalence of no significant (P > 0.05) difference among the age Cryptosporidium. In another study, conducted in categories (Table 3). Romania, the presence of Cryptosporidiosis was observed in 24% of the goat kids [42]. The differences in the DISCUSSION prevalence of Cryptosporidium infections in lambs and goat kids raised in different geographical regions may be Out of 364 fecal samples collected during this study, the result of differences in the levels of contamination of the overall prevalence of Cryptosporidium in all species the environment with oocysts of the parasite or may be was found to be 14%. This result was nearly in agreement due to differences in the infectivity of different with Shoukry et al. [36]. He reported an overall 20% Cryptosporidium spp. populations. It is also possible that prevalence of Cryptosporidiosis in cattle, sheep, goats the quality of hygienic conditions of animal husbandry and buffalos in Egypt. In Iraq, similar finding was reported and grazing practices may have influenced the exposure byal-dabbagh et al. [37]. In Turkey, the prevalence rate of animals to cryptosporidium infection. This was also of 21.05 % in neonatal lambs was reported by Gokce et al. due to difference in management of the animals and in [38]. The prevalence in calves was (13.6%) while the origin of animals because animals for slaughter were 236

usually transported from different areas, intermittent CONCULUSION excretion of Cryptospodrium oocysts, the difficulty in detecting oocysts and the type of study design selected Cryptosporidiosis is an emerging protozoan disease, [ 33]. caused by Cryptosporidium species that can cause Higher stocking rate enhances the infection since gastrointestinal infection in a wide variety of mammals infected calves produce large numbers of oocysts into including human, cattle, sheep, goat, pig and horses confined calf house ensuring a high environmental worldwide. In this study four major factors were found to contamination [43]. In contrast, in the extensively reared be studied with the risk of infection with and traditional husbandry system, calves remain in large Cryptosporidium. These include farm types, animal outdoor paddocks or with dam on pasture, where oocysts species, age category and fecal consistency of the are dispersed on a large surface and are exposed to direct animals sampled. Our study was identified (isolated) in sunlight, which reduces the oocysts viability, resulting in calves, lambs and kids in the study area. The rate of a reduced infection pressure [44]. The infection in sheep isolation of Cryptosporidium statistically significance and goats is also common and severity of clinical (P<0.05) between diarrheic and non diarrheic calves, lambs symptoms varies, the infection often causes death of and kids in the study, but there was no significance lambs and kids [45,46] (also mentioned that in the difference in the prevalence rate of Cryptosporidium extensively reared and traditional husbandry system, between the study farms, animal species, (calves, lambs animals remain at large outdoor paddocks or with dam on and kids) and between different age groups (5 days-6 pasture, where oocysts are dispersed on a large surface months and >6 months-12 months) of the study period. and are exposed to direct sunlight, which reduces the Based on the above findings, the following oocysts viability, resulting in a reduced infection pressure recommendations were forwarded: [47]. The 13.6% prevalence of Cryptosporidium in calves Creation of public awareness on the possible in the current study was compared with previous works. existence of cryptosporidiosis is essential Higher prevalence of17.6% in central Ethiopia [33], 19.2% Further study using molecular technique to identify in Zambia [43], 17.9% in France [44], 35.5% in USA, 33.5% the species of the parasite in Vietnam, 27.9% in UK, 28.5% in Sirlanka, 47.9% in Spain and [ 7], 70% in the US. However, it was higher than the REFERENCES figure reported by Fayer et al. [48] (11.9%) in USA. These differences in the prevalence among countries may be 1. CSA, 2010/11. Central Statics Authority. Report on attributed to the difference in the stocking rate and Livestock and livestock characteristics. Agricultural husbandry system of livestock production system of the sample survey. Vol 2, No. 505. stastical Bulletin. countries. Besides, these variations could also be due to Addis Ababa, Ethiopia. the difference in the susceptibility of the target population 2. Sintayehu, Y., B. Fekadu and T. Azage, 2008. Dairy that related to age difference, diarrheic status and production, processing and marketing system of hygienic practice. Furthermore, the discrepancy between Shashemene-Dilla area, south, Ethiopia. Improving the sensitivity of the diagnostic tests utilized might also productivity and market successof Ethiopian farmers the cause of this variation [43]. project. International Livestock Research Institute, It was also agreed by Xiao et al. [49] and Addis Ababa, Ethiopia. Nguyen et al. [ 50] that although Cryptosporidium was 3. Abdulah, M., R. Acter, S. Cabil, M. Can and observed among all age groups, the prevalence of the M. Abdul, 2013. Characterization of bacterial disease in calves less than 6 months is significantly pathogens isolated from calf diarrhea in panchagarp higher than older cattle. Additionally, some of the distinict of Bangladish. J. Ayrico. Food. Tech., synergic infection of enteropathogens such as Rota virus, 3(6): 8-13. Corona virus, Salmonella and E. coli can result in the 4. Gillhuber, J., D. Rilgamer, K. Pfister and M. Scheuerle, immunocompromised condition and the newborn animal 2004. Giardiasis and other Enteropathogenic will be more susceptible to the Cryptosporidium infection infections a study on diarrheic calves in southern [44]. Germany BMC Res. Notes., 7: 112-120 237

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