Prevalence of Eimeria Species Among Sheep and Goats in Suez Governorate, Egypt ⁎ Walaa I

International Journal of Veterinary Science and Medicine xxx (xxxx) xxx–xxx

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International Journal of Veterinary Science and Medicine

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Full Length Article Prevalence of Eimeria species among sheep and goats in Suez Governorate, Egypt ⁎ Walaa I. Mohamadena,b, , Nahla H. Sallamc, Eman M. Abouelhassanc a Department of Animal Medicine, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt b College of Grassland Science, Gansu Agricultural University, Lanzhou 730070, China c Department of Veterinary Parasitology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt

ARTICLE INFO ABSTRACT

Keywords: Coccidiosis is a disease of high economic importance caused by Eimeria species that show ubiquitous dis- Egypt tribution among several species including small ruminants. The prevalence of Eimeria infection in sheep and Eimeria goats in Geneffe village, Suez Governorate, Egypt was determined during the period from March 2015 to PCR February 2016. Total of 277 animals (142 sheep and 135 goats) were clinically examined and fecal samples Small ruminants were collected and tested both microscopically and by PCR. Sera samples of sheep and goats under 1 year were Subclinical collected for biochemical analysis. Results revealed that (60%) of goats and (57.70%) of sheep were suffering from subclinical coccidiosis. Adult female goats were significantly (P<0.05) more infected (82.2%) than adult male goats (40%). Eimeria infection was significantly prevalent in summer (75%) and autumn (74.2%) in sheep than winter (38.2%) and spring (43.2%), while goats did not show significant seasonal variations of infection. The Eimeria species were identified as E. crandallis, E. granulosa, E. ovina, E. parva, E. faurei, E. ovinoidalis, E intricate, E. pallida, E. arloingi,andE. ahasta in sheep, and E. ninakohlyakimovae, E. hirci, E. caprina, E. christenseni, E. jolchijevi, E. apsheronica and E. arloingi in goats. Although animals were subclinically infected with coccidia, some significant biochemical changes were observed in serum samples of sheep and goats. The molecular detection of Eimeria oocysts did not yield any positive results but after sporulation, Eimeria oocysts were detected at zone 100 bp. Our results showed a moderate prevalence of Eimeria infection among adult and yearling sheep and goats in Geneffe village. Suez governorate, Egypt.Hence, good control and prevention programs are necessary.

1. Introduction subclinical form, impairment of growth is the main sign, and reduced milk production in dairy goats has also been recorded [9]. Under in- Coccidiosis (Eimeriosis sensu stricto) is a protozoan infection caused tensive breeding systems which accompanied by high animal density by parasites of the genus Eimeria that develop and propagate in the and high productivity, coccidiosis may become an infection of sig- small and the large intestines of animals and affect young age parti- nificant economic importance which might lower thriftiness and pro- cularly [1]. Coccidiosis is a serious disease of small ruminants in Egypt ductivity of small ruminants [7]. [2] as well as in various parts of the world and in different animal Surveys based on the examination of ruminant feces have shown species [3–5] which emphasize the importance of further studies for that most animals are infected with a wide variety of Eimeria species better control and prevention programs. from an early age [10,11]. Eimeria diagnosis usually depends on mor- Small ruminants from all ages and breeds are susceptible to Eimeria phological detection of the oocysts by light microscope. That method infection, however, lambs from 3 weeks to 5 months of age are most encompasses several shortcomings including time and effort. It depends severely affected by outbreaks of coccidial infection, while the rest of on the skill and experience of the examiner. Recently, modern mole- the flock might act as carriers [6]. Coccidiosis is clinically characterized cular techniques have been deployed for Eimeria identification in- by diarrhea which can be hemorrhagic in adult sheep [7] while kids or cluding PCR amplification technique [12]. Nevertheless, the main issue lambs suffer from watery diarrhea with clumps of mucous and occa- with the PCR procedures is that the Eimeria oocyst wall is extremely sional changes in the color of feces to yellow or brown [8]. In the robust in addition to the low DNA concentration obtained especially if

Peer review under responsibility of Faculty of Veterinary Medicine, Cairo University. ⁎ Corresponding author at: Department of Animal Medicine, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt. E-mail address: [email protected] (W.I. Mohamaden). https://doi.org/10.1016/j.ijvsm.2018.02.004 Received 9 January 2018; Received in revised form 10 February 2018; Accepted 10 February 2018 2314-4599/ © 2018 Faculty of Veterinary Medicine, Cairo University. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/).

Please cite this article as: Mohamaden, W.I., International Journal of Veterinary Science and Medicine (2018), https://doi.org/10.1016/j.ijvsm.2018.02.004 W.I. Mohamaden et al. International Journal of Veterinary Science and Medicine xxx (xxxx) xxx–xxx

2. Materials and methods

2.1. Study sites

The study was conducted in Geneﬀe village at the Northern border of Suez Governorate (30°14′52.4″ N 32°24′55.1″ E), Egypt which con- stitutes the main village of the rural sector of Suez Governorate. This one-year survey was conducted from the beginning of March 2015 till the end of February 2016. The study was conducted according to ethical guidelines approved by ethics of scientiﬁc research committee, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt.

2.2. Clinical examination and sample collection

Fig. 1. Prevalence of single and mixed infection in relation to the age of sheep and goats. The study was targeting apparently healthy sheep and goats in un- organized farms at different locations in Geneffe village. Animals were routinely released for grazing during the day time and sheltered at Table 1 The prevalence of the different Eimeria spp. in positive sheep and goats. night in small buildings (Partially covered system). Animals were given ad Lib. access to water. Sheep and goats of more than 6 months old were Eimeria species Sheep (n = 82) Goats (n = 81) randomly selected using random sampling procedure from the different farms in Geneffe village from March 2015 to February 2016. Sampling Positive Prevalence % Positive Prevalence % was carried out during the four seasons (Spring, Summer, Autumn, and samples samples Winter) throughout the year. Apparently diseased animals, animals Eimeria ovina 22 26.82% –– receiving treatment or animals in the withdrawal period were excluded. E. parva 15 18.29% –– Fecal samples were collected from 142 sheep and 135 goats. The –– E. pallida 11 13.41% number of sampled animals was calculated using Solvin's depending E. granulosa 12 14.63% –– E. ahasta 25 30.48% –– upon the exact number of animals in each population. The fecal samples E. ovinoidalis 10 12.19% –– were collected once directly from the rectum using gloves and stored at E. faurei 15 18.29% –– 4 °C until being examined. E. crandalis 25 30.48% –– –– E. intricate 6 7.31% fi E. ninakohlyakimovae –– 25 30.86% 2.3. Recovery and species morphological detection and identi cation of E. hirci –– 20 24.69% Eimeria oocysts E. caprina –– 14 17.28% –– E. christenseni 13 16.05% The fecal samples were examined by flotation technique using sa- E. jolchijevi –– 10 12.35% turated saline and oocysts per gram (OPG) quantified using modifica- E. apsheronica –– 13 16.04% E. arloingi –– 30 37.04% tion of McMaster [15]. Each sample was performed in triplicates, and the oocysts number of every sample was expressed using McMaster slide multiplied by the dilution factor (×100) to express the OPG. The the oocysts numbers are low in subclinical cases, thus a successful DNA final results of each sample were obtained using the mean value of three extraction from oocysts is imperative for reliable PCR amplification and independent examinations. After examination, the purified oocysts re- detection of Eimeria [13]. covered from each sample were transferred into 2.5% (w/v) aqueous Subclinical coccidiosis is common among small ruminants [14] and potassium dichromate solution to be sporulated at 26–33 °C in a wet the adverse effects of subclinical coccidiosis on animal health and chamber. The species identification of oocysts was performed based on productivity justify the need for screening. Based on our knowledge the time of sporulation Coudert’s key, and oocysts sizes and mor- there is no study of prevalence of this infection in the rural sector of phology (shape, color, form index, presence or absence of micropyle Suez Governorate, Egypt. Therefore, the purposes of this study were to and its cap, presence or absence of residual, polar and stieda bodies) of investigate the prevalence of Eimeria infection in sheep and goat in the oocysts and sporocysts under 400X magnifications [8,16]. Geneffe village, one of the most important and biggest villages in Suez Governorate, Egypt, and to identify the Eimeria species. Attempts were 2.4. Serum analysis also made to determine the prevalence of infection in relation to age, sex and season, and to elucidate the effect of infection itself on the Five milliliter blood were collected from the jugular vein of the health condition and biochemical parameters. Finally, to adopt a recent young sheep and goats (less than one year) that were divided into 2 approach for the molecular detection of Eimeria oocysts. groups; infected group with Eimeria species and control group which included Eimeria-free animals. Samples were centrifuged at 3000 rpm

Table 2 The prevalence of Eimeria infection in the examined sheep and goats in relation to age and sex.

Animals groups Examined animals Positive No. Prevalence% Males Females

Positive no. Prevalence % Positive no. Prevalence %

Lambs 66 36 54.5 17/30 56.6% 19/36 52.7% Adult sheep 76 46 60.5 13/32 40.6% 37/45 73.3% Kids 60 27 45 15/28 73.3% 12/35 37.5% Adult goats 75 54 72 12/30 40% 37/45* 82.2%*