516 Isrok (Eimeria Species).cdr Eimeria SPECIES COMPOSITION AND FACTORS INFLUENCING OOCYSTS SHEDDING IN DAIRY FARM, BANDUNG, INDONESIA ISROK MALIKUS SUFI , UMI CAHYANINGSIH nd ETIH SUDARNIKA 1* 2 2 a 1 Disease Investigation Center Subang, Ministry of Agriculture, Subang 41212, Indonesia 2 Department of Animal Disease Sciences and Veterinary Public Health, Faculty of Veterinary Medicine, Institut Pertanian Bogor a, Bogor 16680, Indonesi Received 15 July 2015/Accepted 25 May 2017 ABSTRACT Coccidiosis is one of the most widely distributed parasitic diseases of cattle throughout the world. Coccidiosis infection in ruminants was caused by Eimeria spp. The objective of this study was to determine Eimeria species composition and various factors influencing Eimeria oocysts shedding in dairy farm. This study was conducted with a cross-sectional study design in dairy farm in South Bandung District from July 2014 to January 2015. Samples were obtained from 400 dairy cattle (196 cattle at age < 6 months, 37 cattle at age 6 - 12 months and 167 cattle at age > 12 months). Fecal samples were collected, examined and counted for Eimeria species composition and numbers of oocysts per gram of feces (OPG) using McMaster technique. A questionnaire was completed for individual dairy cattle farmer to record information about cattle's health and husbandry. The effect of cattle's sex, age and type of pen flooring to OPG values were analyzed using Mann-Whitney and Kruskal-Wallis tests. The Kruskal-Wallis test was performed followed by Dunn test as a multiple comparison test. Ten species of Eimeria were identified in all infected cattle. Among the Eimeria identified species, Eimeria bovis was found to have the highest prevalence (42.5%), followed subsequently by Eimeria wyomingensis (39.1%), Eimeria bukidnonensis (32.4%), Eimeria pellita (26.3%), Eimeria auburnensis (19.6%), Eimeria zuernii (17.3%), Eimeria cylindrica (3.9%), Eimeria canadensis (3.9%), Eimeria brasiliensis (3.4%) and Eimeria alabamensis (1.1%). The numbers of oocysts shed was correlated significantly (p < 0.05) with cattle's sex and age as well type of pen flooring which influenced the infection pressure. Younger calves aged less than 6 months shed the highest amount of Eimeria oocysts than older cattle. Many factors may cause the increasing number of OPG in fecal samples. Therefore, it is important to keep good sanitation and control of Eimeria among dairy cattle in the KPBS Pangalengan dairy farm. Keywords: Coccidiosis, cross-sectional, dairy farm, Eimeria, OPG INTRODUCTION Coccidiosis is a protozoic disease in cattle caused by Eimeria spp More than twelve different . species of Eimeria have been described in cattle ; most of which are considered harmless. Of these many species, E. bovis and E. zuernii are highly pathogenic causing mortality and morbidity by disturbing absorption mechanisms (Rehman et al. 2011). Infection calves large occurs on caused by numbers of oocyst of E. bovis or E. zuernii may s , result in severe diarrhea with feces containing blood, fibrin and intestinal tissue. However, coccidiosis in cattle commonly occurs as subclinical disease without signs of the disease and involving great economical losses due to reduced appetite, reduced body weight, impaired feed conversion, unthriftness, diarrhea, dysentery, anemia and increased susceptibility to other diseases (Abebe et al. 2008). A poorer body condition score (BCS), potentially as the result of a disease, can be connected to a lower milk production in dairy cattle (Lassen 2009). C is estimated to cause annualoccidiosis economic loss in excess of US 400 million in the US D (Bruhn et al. 2011). The development of clinical coccidiosis in cattle mainly depends on factors such as species * Corresponding author: isrok.sufi@gmail.com 104 BIOTROPIA 4 2 7 104 113 Vol. 2 No. , 201 : - DOI: 10.11598/btb.201 .2 . .7 4 2 516 of , age of infected animal, number of Eimeria oocysts ingested, presence of concurrent infections, as well as type of production system and management practices (Abebe . 2008). et al The aim of this present study was to determine the species composition and factors Eimeria influencing the incidence of oocysts Eimeria shedding in Cooperation of South Bandung Dairy Farm (KPBS) Pangelangan, District of Bandung, Indonesia. Thus, determination of species composition and evaluation of factors influencing the shedding of oocysts per Eimeria gram (OPG) is very useful in designing efficient control strategies. MATERIALS AND METHODS Study Area T he study was conducted in KPBS Pangalengan, South Bandung District, West Java Province from July 2014 to January 2015. KPBS Pangalengan Bandung covers 27.3 ha of highland area with an altitude of 1,000 to 1,420 m asl, 12 – o 28 C mean annual temperature and 60 – 70% relative humidity. The average annual rainfall was > 2,000 mm during the study period. Study Design C applied based ross-sectional study design was on Thrusfield (2007). A total of 400 dairy cattle (n) from a total population of 14 000 dairy cattle , distributed on 37 cooperation service area (TPK) and 225 farmers group were selected for the s study with 95% confidence interval, 50% expected prevalence (P) and 5% accepted error (L). The relevant formula for a 95% confidence interval is Thrusfield 2007) based on ( : n = 4PQ/L 2 where: n = total of cattle P = expected prevalence Q = 1 – P L = accepted error Samples obtained from dairy cattle farmers covered all age groups i.e. calves (age < 6 months); weaners (6 - 12 months); and adults (age > 12 months). From the total of 225 farmers groups, 10 farmers groups consisted of 80 farmers were randomly selected because these farmers had complete cattle age groups. From each of these farmers, five fecal cattle samples were collected including all calves in the selected farmers, while the remainder fecal samples were taken from weaners and adults. Sample Collection Fecal samples of 20 - 50 g were collected from each age group. The feces was directly obtained from cattle's rectum or immediately collected after cattle defecation. Fecal samples were stored in respective plastic bag and preserved in o refrigerator at 4 C before being tested in the laboratory for the occurrence of Eimeria spp. oocysts. Fecal consistency was assessed immediately after sampling and classified as normal or diarrheal without any additional differentiations (Bangoura et al. 2011). The plastic bags containing fecal samples were labeled with sampling information such as sample number, sampling date, sampling location, cattle's age group, and farmer's name. Practices of animal health management were recorded by the participating farmers in a questionnaire. In addition, record of cattle's sex and age (in months) were documented for each cattle from which a sample was taken. Counting of POocysts er Gram of Feces (OPG) Fecal samples were examined and oocysts numbers were counted using the McMaster method. Two grams of fecal material was mixed thoroughly with 28 mL of sugar-salt solution and filtered through a 200 µm mesh wire sieve or tea strainer. The suspension was equally poured into two McMaster counting chambers (2 x 0.15 mL) and was let still for 5 minutes. The counting was carried out using light microscope with 100x magnification. The numbers of oocysts in the two chambers were multiplied by dilution factor (50) to obtain the number of oocysts per gram of feces (OPG). This protocol was modified from Dong et al. (2012), Lucas et al. (2006) and Soulsby (1986). Eimeria Species Identification The remainder of each positive samples were centrifuged at 1,500 rpm for 5 minutes at room temperature of 25 C For identification, a o . Eimeria species composition and factors influencing oocysts shedding in dairy farm – Sufi et al. 105 Data Analysis The effect of cattle's sex and age as well as type of pen flooring to Eimeria OPG counts were analyzed using Mann-Whitney and Kruskal- Wallis tests. The Kruskal-Wallis test was performed followed by Dunn test as multiple comparison test. RESULTS AND DISCUSSION Species Identification and Composition of Eimeria. Results of this study showed that the shedding of Eimeria oocysts were commonly occurred in KPBS Pangalengan dairy farm in Bandung. From flotation of the oocysts in sugar-salt Eimeria solution was performed. Oocysts were measured under ocular eye piece that was calibrated with micrometer under 40x objective lense of a light microscope. Identification of species was Eimeria based on the morphological features of the oocysts (size, index, shape, color and texture of oocyst's wall, presence or absence of micropyle and polar cap) with the aid of taxonomic keys (Soulsby 1986; Daugschies & Najdrowski 2005). Measurement of oocysts index value were performed by dividing the length and width of oocysts. The shape of oocysts Eimeria were examined for each sample and classified as round (length/width = 1), ovoid (length/width between 1 - 1.5) and ellips (length/width > 1.5) (Cahyaningsih & Supriyanto 2007). 106 BIOTROPIA Vol. 24 No. 2, 2017 (a) (b) (c) (d) (e) (f) (g) (h) (i) (j) 10 µm10 µm 10 µm 10 µm 10 µm 10 µm 10 µm 10 µm 10 µm 10 µm Figure 1 The results of Eimeria species oocysts in dairy cattle in KPBS Pangalengan, Bandung (a) Eimeria bovis; (b) Eimeria wyomingensis; (c) Eimeria bukidnonensis; (d) Eimeria pellita; (e) Eimeria auburnensis; (f) Eimeria zuernii; (g) Eimeria cylindrica; (h) Eimeria canadensis; (i) Eimeria brasiliensis; (j) Eimeria alabamensis 400 fecal samples, there were 179 fecal samples (44.8% prevalence; Confidence Interval (CI) 95%; 40.0 49.6 ) contained Eimeria oocysts. A total of - 10 Eimeria species i.e. E. bovis, E. wyomingensis, E. bukidnonensis, E. pellita, E. auburnensis, E. zuernii, E. cylindrica, E. canadensis, E. brasiliensis and E. alabamensis were identified from the 179 positive fecal samples collected from KPBS Pangalengan dairy farm (Fig. 1, Table 1, Table 2). Our study showed that E. bovis was the most prevalent species (42.5%; CI 95%: 37.1 - 47.8) and had the OPG highest mean OPG number (538.2 ( ocysts er ram of feces); CI 95%: 191.5 -O P G 884.8) followed by (39.1; CI 95%: E. wyomingensis 33.8 - 44.4 and 285.0; CI 95%: 136.0 - 434.0) and E. bukidnonensis (32.4; CI 95%: 27.3 - 37.5 and 237.1; CI 95%: 128.3 - 345.9) the . E. zuernii had second highest mean OPG counts (509.7; CI 95%: 46.1 - 973.3). The lowest prevalent species was E. alabamensis (1.1; CI 95%: 0.0 - 2.3 and 75.0; CI 95%: 0.0 - 392.7) (Table 3). A number of authors reported that E. bovis was the most prevalent species in cattle, but clinical coccidiosis was not observed in the calves or in adults cattle (Kennedy & Kralka 1987; Lucas et al. 2006; Heidari & Gharekhani 2014). The results of those studies were in close agreement with the results of our study. According to Waruiru et al. (2000), the mere present of pathogenic Eimeria spp. did not necessarily indicate clinical disease. Eimeria species were classified as highly pathogenic (E. bovis and E. zuernii), low pathogenic (E. ellipsoidalis, E. alabamensis, E auburnensis and E. subspherica) and non-pathogenic (E. brasiliensis, E. bukidnonensis, E. canadensis, E. cylindrica, E. pellita dan E. wyomingensis) (Lassen & Jarvis 2009). In other tropical area such as Brazil, s several species of Eimeria in cattle found were also ( Floriao et al. 2015). E. bovis was recorded as the highest prevalen coccidian species which s in t wa accordance with reports of Arslan and Tuzer (1998), Lucas et al. (2006) and Abebe et al. (2008). 107 Table 1 Oocyst morphology of Eimeria species of cattle has been identified in fecal samples obtained from KPBS Pangalengan, Bandung Length (µm) Width (µm) Index Form Color Other characteristics Eimeria species 26.25-33.00 18.75-22.50 1.17-1.48 Ovoid Greenish-brown Bilayered wall, micropyle E. bovis *29.77±1.37 *21.58±0.73 *1.38±0.05 37.50-43.50 26.25-30.75 1.22-1.49 Ovoid Greenish-brown Single layered wall, micropyle E. wyomingensis *42.67±2.04 *29.58±1.20 *1.44±0.04 47.25-49.50 33.75-37.50 1.26-1.47 Ovoid Yellowish -brown Bilayered wall, micropyle E. bukidnonensis *48.70±0.68 *35.72±1.88 *1.37±0.08 36.00-37.50 26.25-29.25 1.26-1.37 Ovoid Dark brown Thick wall, micropyle E. pellita *37.43±0.28 *29.16±0.51 *1.28±0.02 37.50-45.75 22.50 1.67-2.03 Ellips Yellowish -brown Bilayered wall, micropyle E. auburnensis *38.74±2.73 *22.50±0.00 *1.72±0.12 15.75-20.25 15.00-17.25 1.05-1.35 Ovoid Pale yellow Thin wall, no micropyle E. zuernii *19.87±1.19 *16.65±1.01 *1.20±0.08 25.50-27.00 15.00 1.70-1.80 Ellips Pale yellow Single layered wall, no micropyle E. cylindrica *25.93±0.73 *15.00±0.00 *1.73±0.05 30.00-33.75 25.50-26.25 1.14-1.32 Ovoid Yellowish Bilayered wall, micropyle E. canadensis *30.42±1.25 *26.00±0.38 *1.17±0.06 41.25-42.75 25.50-26.25 1.60-1.63 Ellips Brownish-yellow Bilayered wall, micropyle E. brasiliensis *41.50±0.53 *25.67±0.33 *1.62±0.007 15.00-18.75 11.25-15.00 1.25-1.33 Ovoid Pale yellow Smooth wall, no micropyle E. alabamensis *16.88±2.65 *13.13±2.65 *1.29±0.06 Note: * mean; ± standard deviation = = Eimeria species composition and factors influencing oocysts shedding in dairy farm – Sufi et al. 108 BIOTROPIA Vol. 24 No. 2, 2017 T ab le O o cy st m o rp h o lo g y o f s p ec ie s o f c at tl e b as ed o n r ef er en ce s 2 E im er ia S p ec ie s A cc o rd in g to S o u ls b y (1 9 8 6 ) A cc o rd in g to D au gs ch ie s an d N aj d ro w sk i (2 0 0 5 ) L en gt h (µ m ) W id th (µ m ) In d ex F o rm C o lo r O th er ch ar ac te ri st ic s L en gt h (µ m ) W id th (µ m ) In d ex F o rm C o lo r O th er ch ar ac te ri st ic s E . bo vi s 2 3 -3 4 1 7 -2 3 1 .3 5 -1 .4 8 O v o id G re en is h - b ro w n S m o o th w al l, m ic ro p yl e 2 3 -3 4 1 7 -2 3 1 .3 5 -1 .4 8 O v o id B ro w n is h - ye ll o w B il ay er ed w al l, m ic ro p yl e E . w yo m in ge ns is 3 7 -4 4 .9 2 6 .4 -3 0 .8 1 .4 0 -1 .4 6 O v o id G re en is h - b ro w n M ic ro p yl e 3 7 -4 5 2 6 -3 1 1 .4 2 -1 .4 5 O v o id Y el lo w is h - b ro w n S in gl e la ye re d w al l, m ic ro p yl e E . bu k id no ne ns is 4 7 -5 0 3 3 -3 8 1 .3 2 -1 .4 2 O v al Y el lo w is h - b ro w n B il ay er ed w al l, m ic ro p yl e 4 7 -5 0 3 3 -3 8 1 .3 2 -1 .4 2 P ea r- sh ap ed B ro w n is h - ye ll o w B il ay er ed w al l, m ic ro p yl e E . pe lli ta 3 6 .1 -4 0 .9 2 6 .5 -3 0 .2 1 .3 5 -1 .3 6 E g g- sh ap ed D ar k b ro w n T h ic k w al l, m ic ro p yl e 3 6 -4 1 2 6 -3 0 1 .3 7 -1 .3 8 O v o id D ar k b ro w n T h ic k w al l, m ic ro p yl e E . au bu rn en si s 3 2 -4 6 2 0 -2 5 1 .6 0 -1 .8 4 E ll ip s Y el lo w is h - b ro w n S m o o th w al l, m ic ro p yl e 3 2 -4 6 2 0 -2 5 1 .6 0 -1 .8 4 E ll ip so id Y el lo w is h - b ro w n B il ay er ed w al l, m ic ro p yl e E . zu er ni i 1 5 -2 2 1 3 -1 8 1 .1 5 -1 .2 2 S u b - sp h er ic al P al e ye ll o w T h in w al l, n o m ic ro p yl e 1 5 -2 2 1 3 -1 8 1 .1 5 -1 .2 2 S u b o vo id C o lo rl es s S in gl e la ye re d w al l, n o m ic ro p yl e E . cy lin dr ic a 1 6 -2 7 1 2 -1 5 1 .6 7 -1 .8 0 C yl in d ri ca l C o lo rl es s T h in w al l, n o m ic ro p yl e 1 6 -2 7 1 2 -1 5 1 .6 7 -1 .8 0 E li p so id C o lo rl es s S in gl e la ye re d w al l, n o m ic ro p yl e E . ca na de ns is 2 8 -3 7 2 0 -2 7 1 .3 7 -1 .4 0 O v o id / el li p s Y el lo w is h - b ro w n S m o o th w al l, m ic ro p yl e 2 8 -3 7 2 0 -2 7 1 .3 7 -1 .4 0 O v o id / el li p so id Y el lo w is h B il ay er ed w al l, m ic ro p yl e E . br as ili en si s 3 4 .2 -4 2 .7 2 4 .2 -2 9 .9 1 .6 4 -1 .6 5 E ll ip s Y el lo w S m o o th w al l, m ic ro p yl e 3 4 -4 3 2 4 -3 0 1 .6 5 -1 .6 7 E ll ip so id B ro w n is h - ye ll o w B il ay er ed w al l, m ic ro p yl e E . al ab am en si s 1 3 -2 4 1 1 -1 6 1 .1 8 -1 .4 4 P ea r- sh ap ed C o lo rl es s T h in w al l, n o m ic ro p yl e 1 3 -2 4 1 1 -1 6 1 .1 8 -1 .4 4 O v o id C o lo rl es s / p al e ye ll o w T h in w al l, n o m ic ro p yl e According to Daugschies Najdrowski (2005), and E. bovis and E. zuernii are the most pathogenic of the bovine coccidia urthermore, E. alabamensis . F causes disease at extremely large infective doses (> 10 million oocysts) only. E. alabamensis usually induces water y diar rhea without blood, dehydration, depression and reduced growth , whereas infection remains subclinical under moderate infection pressure. Single or Mixed Infection -Species Caused by Different peciesEimeria S In this study, mixed-species infection experienced by a single cattle hosting several Eimeria species were commonly observed. The number of Eimeria species in single and mixed- species infection per examined fecal sample ranged from 1 to 5. Mixed-species infection caused by 2 to 5 Eimeria species were found in 55.9% of cases (CI 95%; 50.5 - 61.3) The . remainder fecal samples had been infected with single infection of Eimeria species (44.1%; CI 95%: 38.7 - 49.5) (Table 4). theThis finding is similar to works of Kennedy Kralka (1987) in Canada, which and reported 5 species. The results of mixedEimeria - species we infection re lower than the observation of Arslan and Tuzer (1998) in Turkey who reported 6 species, Abebe . (2008) in Eimeria et al Ethiopia who reported 7 species Yu Eimeria , et al. (2011) in China who reported 10 Eimeria species and Dong (2012) who recorded et al. also in China 8 species. Many previous studies indicated Eimeria that under natural conditions, mixed-species infection re much more common than cases we single species infection (Yu 2011; Dong et al. et al. 2012). The level of pathogenicity of Eimeria species increased higher level due to mixedto a - species Eimeria, infection with other pathogenic causing mortality in cattle (Daughschies & Najdrowski 2005). 109 Table 3 Eimeria species identified, the prevalence and mean OPG counts in dairy cattle in KPBS Pangalengan dairy farm, Bandung Species Dairy cattle (n = 179) Mean OPG (CI 95%)Positive no. Mean Prevalence (%) (CI 95%) E. bovis 76 42.5 (37.1 - 47.8) 538.2 (191.5 - 884.8) E. wyomingensis 70 39.1 (33.8 - 44.4) 285.0 (136.0 - 434.0) E. bukidnonensis 58 32.4 (27.3 - 37.5) 237.1 (128.3 - 345.9) E. pellita 47 26.3 (21.5 - 31.1) 177.7 (124.7 - 230.7) E. auburnensis 35 19.6 (15.2 - 23.9) 397.1 (0.0 - 820.9) E. zuernii 31 17.3 (13.2 - 21.4) 509.7 (46.1 - 973.3) E. cylindrica 7 3.9 (1.8 - 6.02) 78.6 (42.2 - 115.0) E. canadensis 7 3.9 (1.8 - 6.02) 121.4 (51.5 - 191.3) E. brasiliensis 6 3.4 (1.4 - 5.3) 125.0 (45.4 - 204.6) E. alabamensis 2 1.1 (0.0 - 2.3) 75.0 (0.0 - 392.7) Table 4 Single and mixed infection of Eimeria species No. of Eimeria spp. Dairy cattle (n = 179) Positive no. % of positive samples (CI 95%) 1 79 44.1 (38.7 - 49.5) 2 59 33.0 (27.8 - 38.1) 3 24 13.4 (9.7 - 17.1) 4 15 8.4 (5.4 - 11.4) 5 2 1.1 (0.0 - 2.3) Total 179 100.0 Eimeria species composition and factors influencing oocysts shedding in dairy farm – Sufi et al. The Effect of Sex, Age and Type of Cattle's Pen Flooring to OPG CountsEimeria The effect of various categorical factors to OPG counts were first analyzed with the Mann- Whitney and Kruskal-Wallis tests. A Mann- Whitney test was used to compare mean OPG between 2 groups, whereas Kruskal-Wallis test was used to compare mean OPG among 3 or more groups. The Kruskal-Wallis test was performed followed by Dunn test as a multiple comparison test. A p value < 0.05 was required to indicate significance. Several categorical factors (cattle's sex, age and type of pen flooring) and mean OPG counts are presented in Table 5. A highly significant ( = 0.000) effect was p observed between different sexes of cattle . Intensity of infections were shown by Eimeria mean OPG found in cattle. This study showed that higher OPG counts were found in male cattle (855.6 OPG; CI 95%: 150.9 – 1 560.4) compared , with females (144.5 OPG; CI 95%: 77.7 211.4). - In this study, male cattle harbored more coccidia than female cattle. This situation might be caused by less care given to the male cattle as compared to the female cattle that were deemed to produce future cows. This result was consistent with reports from other researchers showing significant correlation ( < 0.05) between cattle's p sex and coccidiosis infection (Heidari & Gharekhani 2014). Previous studies done on cattle reported higher prevalence of in female Eimeria than in male cattle. Nevertheless, this could be attributed to the greater physiological stress experienced by female cattle in relation to pregnancies and breeding as compared to male cattle (Rehman 2011; Dawid 2012; et al. et al. Alemayehu 2013; Heidari 2014).et al. et al. R is shed esults of th study showed that calves much higher numbers of oocyst than s those of weaners and adults. For instance, the mean OPG for - the calves was 537.0 (CI 95%; 231.8 842.2), whereas the weaners and adults mean OPG for were 154.1 (CI 95%; 79.8 228.3) and 22.5 (CI - 95%; 11.0 33.9), respectively. Analysis showed - that there were statistically significant differences in OPG levels among different age groups (

0.05) the calves in OPG p to levels caused by coccidiosis (Table ).5 Age is a major risk factor in coccidiosis spreading orbidity and risk of infection , while m s s are gr ater in calves (Abebe 2008)e et al. compared to other age groups. Calves had significantly higher oocyst counts ( < 0.05) than adultss p . These results were et in agreement with Waruiru al. et al. (2000) in Kenya and Dong (2012) in Shanghai, China who demonstrated that age strongly influence the intensity of OPG d Eimeria counts in cattle. Coccidiosis is a self-limiting disease. Spontaneus recovery without specific treatment is common when multiplication stage of the coccidia has passed, which suggests that previous exposure may have contributed to the 110 BIOTROPIA Vol. 24 No. 2, 2017 Table 5 Results of Mann-Whitney and Kruskal-Wallis tests followed by Dunn test toward several factors and mean OPG counts Categorical factors No. Dairy cattle (n) Mean OPG (CI 95%) p value Sex Male 80 855.6 (150.9 – 1,560.4) 0.000* Female 320 144.5 (77.7 - 211.4) Age Calves (< 6 months) 196 537.0a (231.8 - 842.2) 0.000* Weaners (6 -12 months) 37 154.1a (79.8 - 228.3) Adults (> 12 months) 167 22.5b (11.0 - 33.9) Type of pen flooring Straw 13 53.9a (0.0 - 162.4) 0.000* Wood 70 703.6b (0.0 – 1,450.2) Rubber 96 405.7b (116.9 - 694.6) Cement 221 116.7a (52.4 - 181.1) development of a certain immunity level in the older cattle as compared to the younger ones that did not have previous exposure (Dong 2012; et al. Heidari 2014; Kocis 2015).et al. et al. This study showed that w Eimeriaeaners shed oocysts OPG counts of 154.1 (CI 95%; 79.8with - 228.3). ean OPG the weaners re lower M for we than the calves ( > 0.05). In tudy, that for p this s range of were 0 - OPG in calves and weaners 24 450 and 0 900, respectively. OPG values over , - 5 000 indicate a clinical case (Arslan Tuzer , & 1998). esults of oocysts counts calves R Eimeria for and weaners in were not different from this study previous Bruhn (2012) wh study by et al. ich reported that no difference was observed ( >p 0.05) in EPG and OPG counts between calves in the pre-weaning and post-weaning phases. Most cattle examined during this study had low OPG counts, suggesting that the infections were usually subclinical. This result concurred with other cross-sectional observational studies on Eimeria spp. in Iran, which also did not observe cases of clinical cases of clinical coccidiosis among infected cattle, probably due to low quantities of oocysts eliminated in the cattle's feces (Heidari 2014; Heidari & Gharekhani et al. 2014). L Eimeriaevel of oocyst shedding was significant depende on different types of pen ly d flooring ( < 0.05). When the types of flooring p were compared, on straw floorcattle kept ing exhibited lower OPG values (53.9; CI 95%: 0.0 - 162.4) than on wood those kept on flooring (703.6; CI 95%: 0.0 – 1 450.2) and rubber , on floor (405.7; CI 95%: 116.9 - 694.6) ( < 0.05). ing p However, straw floor significanting did not show difference on p compared to OPG counts ( > 0.05) cement floor . Moreover, wood floor ing ing did not show difference on significant OPG counts compared to ing p rubber floor ( > 0.05). Yet, Eimeria p on OPG counts was higher ( < 0.05) wood floor compared to cement floor . ing ing Higher oocysts count was recorded in Eimeria cattle ing reared on rubber floor compared to cement floor ( < 0.05) (Table ).ing p 5 This finding was in agreement with a study by Bangoura et al. (2011) who described a significant association between different flooring types used for rearing cattle and the level of Eimeria oocysts shedding ( < 0.05). Rehman et al. (2011) stated p that Eimeria infection was more prevalent ( < 0.05) in non-cement flooring type compared p to partially cement flooring type. Lower Eimeria prevalence in cattle kept on cement flooring may be caused by the easiness of cleaning and disinfecting cement flooring, which resulted to less contaminated floor compared to non-cement floor. In this study the Eimeria, lowest level of oocysts in cattle kept on straw occurred flooring than those kept on other flooring types, because solid floor (wood, rubber and cement ing types ) are ier the solid flooring dirt than straw floors if types are S not scraped and cleaned properly. traw floor type usually used by calves ing is temporarily aged 0 to 1 month to keep warm. the calves Daily r al ement of ingemov and replac straw floor with a new straw resulted in lower OPG set of s Eimeria counts occurence of and reduced the Eimeria oocyst shedding. revious studies concluded that P cleaning s an important factor in preventing wa high of oocysts (Bangoura counts Eimeria et al. 2011; Rehman 2011).et al. was cattle Clinical coccidiosis observed in pens having leadinginsufficient aeration to high concentration of ammonia, CO and moisture. 2 Also, clinical coccidiosis occurred in cattle pens where feces accumulated on the ground due to unsuited slatted floors (Daughschies & Najdrowski 2005). Local temperature, humidity, ammonia concentration and pH on pen flooring affected ' et al. oocysts survival (Gulliksen 2009; Bangoura 2011).et al. In having intensive rearing high population density, disease transmission more readilyis occurred and oocysts are highly available within the environment. Thus, in dairy calves, coccidiosis occurs more frequently and appears with greater severity (Bruhn 2011). It can be et al. assumed that the in a group will be cattle living infected at the same time, or at around the same time, under field condition. Moreover, continuous oocysts shed from subclinical ding infected calves contaminate the environment and the caus severe coccidiosis in cattle's hair ing highly susceptible new calves that are kept in these areas (Abebe 2008; Alemayehu 2013). et al. et al. Different hygiene conditions and far m management, breeding, study design and animal s methods, climates and different geographical regions may be the main cause of varied results (Yu 2011)et al. . 111 Eimeria species composition and factors influencing oocysts shedding in dairy farm – Sufi et al. CONCLUSIONS E. bovis was the highest prevalent Eimeria species and had the highest level of mean OPG compared to other Eimeria spesies. Mixed infections in single cattle with 2 – 5 Eimeria species were commonly observed. Several species of Eimeria in cattle are found in other tropical areas. Various categorical factors (cattle's sex, age and type of pen flooring) influenced the number of OPG found in fecal samples. Eimeria has significant pathogenic potential. It is important to control the occurrence of Eimeria. 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