Global Veterinaria 9 (6): 700-705, 2012 ISSN 1992-6197 © IDOSI Publications, 2012 DOI: 10.5829/idosi.gv.2012.9.6.66105

Comparative Study of Hindgut Ciliates in , and Donkeys in Turkey

Gözde Gürelli

Department of Biology, Faculty of Science and Art, Kastamonu University, 37100 Kuzeykent/Kastamonu, Turkey

Abstract: The ciliate fauna in the hindgut of 24 equids (8 horses, 8 mules, 8 donkeys) living in the same area was surveyed. Thirty four species belonging to 21 genera were identified. It was possible to identify ciliate protozoa, including 28 species to 18 genera in horses, 27 species to 16 genera in mules and 20 species to 16 genera in donkeys. The average number of ciliate species was 12.8±3.0 in horses, 8.1±5.8 in mules and 7.5±4.0 in donkeys. One of the mules and one of the donkeys had no ciliates. The average density of ciliates was 15.4±10.8 in horses, 6.4±4.6 in mules, 10.4±13.6 in donkeys. This is the second report on the hindgut cilites of mules.

Key words: Hindgut Ciliate Turkey

INTRODUCTION MATERIALS AND METHODS

Equids (horses, donkeys, mules and ) are Fecal samples were collected from 8 horses, 8 mules monogastric herbivores and they have four major sections and 8 donkeys located in the vicinity of Izmir in Turkey. of the gastrointestinal tract. These are the mouth, The samples were collected from February 2012 to July stomach, small intestine and a highly developed large 2012. The fecal samples were collected immediately after intestine, consisting of the caecum, large colon, small defecation and fixed and stained with methylgreen colon and rectum, which are collectively known as the formalin saline solution (MFS) as their original volume hindgut [1, 2]. Hindgut of the equids is inhabited by a [18, 19, 23]. This procedure was used to preserve the unique ciliate fauna which help in the digestion of integrity of the cell and its internal structures. The MFS cellulose and starch [3]. served as a nuclear stain and Lugol’s iodine was used to The intestinal ciliate fauna of equids was first stain skeletal plates. Fecal samples were passed through described by Gruby and Delafond [4]. Since that time, 2.56 mm mesh gauze and kept in the dark until many investigations have been reported [5-20]. The examination. Details of the ciliate morphology were ciliates invade the equid hosts by oral ingestion studied at 100x magnification using an oil immersion (coprophagy) and become established in its large objective microscope. intestine [21]. Total cell counts were made at 400x magnification Although many investigations have been with a Neubauer hemocytometer counting chamber. The conducted on the ciliate fauna of horses, only a few number of cells per 1 ml of intestinal contents can be have been conducted on the ciliate fauna of mules calculated by the following Formula: N= (10/4) x a x d and donkeys [15, 17, 22]. The aim of this study was to (N: number of ciliates per 1 ml of intestinal contents, a: identify and quantify the fecal ciliate fauna of mules, number of ciliates in 4 divisions on the Neubauer donkeys and horses living near the same area hemocytometer, d: sample dilution) [18]. (Izmir-Turkey), comparing the data between each animal Differential counts of species were estimated from group and with previous studies on equids from various smear slides with a total of 150 to 200 cells identified for other locations. each species [18, 19].

Corresponding Author: Gözde Gürelli, Department of Biology, Faculty of Science and Art, Kastamonu University, 37100 Kuzeykent/Kastamonu, Turkey. 700 Global Veterinaria, 9 (6): 700-705, 2012

Classification and identification of species were donkeys living in the present study (10.4±13.2 x10 41 ml ) based on previously published species descriptions and is higher than Cypriot wild donkeys [17] (Table 4). taxonomic lists [6-8, 15, 24, 25]. These variations could result from differences both in the host animals and feed. RESULTS AND DISCUSSION The average ciliate density in the mules living in Turkey wasn’t compared with the mules in China, because Frequency of appearance (i.e. the number of hosts in the only previous report on the fauna of Chinese mules which the species was detected/number of hosts did not give density values [22]. examined x 100) and the relative composition of genera If the ciliate density is compared among Equid and species according to equid hosts are shown in species in Turkey, the average ciliate density in horses is Table 1. 34 species belonging to 21 genera was identified higher than either mules or donkeys. The average ciliate in the 24 equids. The ciliate fauna consisted of 1 genus density is the lowest in mules (Table 3). These hosts are and 1 species of Paraisotrichidae, 8 genera and 17 species living in the same area, but feeding habits are not the of Buetschliidae, 4 genera and 6 species of same. Blepharocorythidae, 2 genera and 3 species of Donkeys characteristically get by on less food than Cycloposthiidae, 5 genera and 6 species of Spirodiniidae a horse of similar size and need more fibre and less protein and 1 genus and 1 species of Allantosomatidae. It was in their diets than horses. In addition this, donkeys are possible to identify ciliate protozoa, including 28 species grazers as well as browsers [26]. Donkeys can graze to 18 genera in horses, 27 species to 16 genera in mules coarser pasture than a horse. Donkeys are fed coarse and 20 species to 16 genera in donkeys. herbage, marsh grass, young thistles and shrubs in their The average number of ciliate species per animal was pasture, feeds that most horses will not eat. Mules are 12.8±3.0 in horses, 8.1±5.8 in mules and 7.5±4.0 in donkeys also very hardy and generally need less concentrated feed (Table 2). It was higher in horses than others. One of than horses. If the mules are fed concentrates, they only mules and one of the donkeys had no ciliates. The species need about half of what a horse requires. Mules like B. postciliata, B. angusta and B. microcorys were present donkeys, need less protein and more fiber than horses in all horses In the 7 donkeys with protozoa, the species [27, 28]. B. postciliata and B. curvigula occurred in all animals. Food type differences are evident in the animals Although no single species was present in all 7 of the feces. Donkeys feces are firmer and drier than that of faunated mules, Triadinium caudatum was present in horses. The lower ciliate density and diversity in the feces 6 animals (Table 1). of the donkeys could be caused by the reduced water The average density of ciliates was 15.4±10.8 in contents of their feces. Because donkeys can survive with horses, 6.4±4.6 in mules, 10.4±13.2 in donkeys (Table 3). little water than other equid species [29]. When the feces It was higher in horses than others. are firm and dry, most protozoa became partially In the present investigation, 34 ciliate species desiccated and destroyed [30]. Besides, the length of the representing 21 genera were identified, but no new hosts’s intestine also plays a role; donkeys have shorter species were detected. This is the first report on the and thicker intestines than horses [17]. hindgut ciliates in mules and donkeys living in Turkey. Spirodinium uncinucleatum, Sulcoarcus It is the second report on the hindgut ciliates of mules in pellucidulus, Spirodinium confusum and Ditoxum the world. In general, the ciliate composition of the equids gravinucleatum were firstly reported from mules in examined in this study is similar to other reports from China [22]. In this study these species were not observed around the world (Table 4). from mules and other equid species. According to When compared with ciliate surveys from other Strelkow [7] Ditoxum gravinucleatum is the same species horses living in various countries, the average ciliate with D. brevinucleatum which was reperted from zebras density in the hindgut of domestic horses (15.4±10.8 x10 4 [31]. S. uncinucleatum and D. brevinucleatum were ml 1) was considerably less than that of kiso horses [14], reported from Cypriot wild donkeys [17], but they were riding horses [12] and racing horses [19]. On the other not observed from domestic donkeys in this investigation. hand, concentrations are higher than those in the other Strelkow [7] classified Cycloposthium edentatum horses examined, the light horses [9], race horses [10], into four morphotypes based on its cell surface and tokara pony [13], Turk rahvan horse [18] and Cypriot its cell size. In this investigation, only one morphotype domestic horses [20]. The average ciliate density in the (C. edentatum) was observed.

701 Global Veterinaria, 9 (6): 700-705, 2012

Table 1: Frequency of appearance and percentage composition of hindgut ciliates in the feces of 24 equids Horses Mules Donkeys ------Familia/genus/ species F(%) P (Mean±SD) (%) F(%) P (Mean±SD) (%) F(%) P (Mean±SD) (%) Paraisotrichidae Paraisotricha 12.5 0.3±0.9 - - - - Colpoidea 12.5 0.3±0.9 - - - - Buetschliidae Bundleia 100 30.5±9.5 75 22.4±14.8 87.5 18.4±11.6 Postciliata 100 9.4±3.6 62.5 9.7±9.7 87.5 11.5±7.2 Piriformis 25 0.9±1.8 25 1.7±3.2 - - Nana 50 1.6±2.0 25 1.5±2.9 25 2.4±4.5 Elongata 75 8.0±5.7 37.5 2.3±3.5 - - Triangularis 75 5.6±5.3 25 2.0±3.8 25 1.0±1.8 Dolichosoma 37.5 3.1±4.9 12.5 1.0±2.9 - - Inflata - - 37.5 4.2±6.9 25 5.4±10.5 Didesmis - - 12.5 0.3±1.0 37.5 1.0±1.7 Ovalis - - 12.5 0.3±1.0 37.5 1.0±1.7 Polymorphella 75 3.6±2.7 37.5 2.5±7.2 50 6.0±7.2 Ampulla 75 3.6±2.7 37.5 2.5±7.2 50 6.0±7.2 Blepharoconus - - 12.5 - 25 1.2±2.3 Hemiciliatus - - - - 12.5 0.7±2.1 Benbrooki - - 12.5 0.6±1.6 12.5 0.5±1.4 Holophryoides 37.5 4.0±6.8 50 7.4±12.8 75 8.9±8.04 Macrotricha 37.5 3.1±5.0 50 7.4±12.8 75 8.9±8.04 Ovalis 12.5 0.9±2.7 - - - - Hemiprorodon 12.5 0.1±0.3 37.5 0.5±0.8 12.5 1.0±2.9 Gymnoposthium 12.5 0.1±0.3 37.5 0.5±0.8 12.5 1.0±2.9 Prorodonopsis 37.5 0.7±1.3 - - - - Coli 37.5 0.7±1.3 - - - - Blepharoprosthium 50 1.1±1.3 37.5 1.5±2.9 12.5 0.5±1.5 pireum 50 1.1±1.3 25 0.4±0.8 12.5 0.5±1.5 Polytrichum - - 12.5 1.0±3.0 - - Blepharocorythidae Blepharocorys 100 33.4±15.7 75 29.8±26.0 87.5 28.5±18.4 Curvigula 75 10.1±8.6 62.5 8.5±8.5 87.5 18.7±10.7 Angusta 100 13.3±4.5 25 2.5±4.7 50 9.8±11.9 Microcorys 100 13.8±7.0 50 18.8±27.9 37.5 5.1±7.3 Ochoterenaia 37.5 3.0±4.3 12.5 1.8±5.0 - - Appendiculata 37.5 3.0±4.3 12.5 1.8±5.0 - - Circodinium 25 1.2±2.1 12.5 1.4±4.0 - - Minimum 25 1.2±2.1 12.5 1.4±4.0 - - Charonina - - 12.5 0.5±1.3 - - Equi - - 12.5 0.5±1.3 - - Cycloposthiidae Cycloposthium 75 9.0±9.8 37.5 4.5±7.2 50 6.3±9.7 Bipalmatum 62.5 7.5±9.0 12.5 0.9±2.6 37.5 5.5±10.0 Edentatum 25 1.5±2.9 25 3.6±7.3 12.5 0.8±2.2 Tripalmaria 25 0.7±1.4 - - - - Dogieli 25 0.7±1.4 - - - - Spirodiniidae Cochliatoxum 25 0.2±0.4 - - 12.5 0.2±0.6 Periachtum 25 0.2±0.4 - - 12.5 0.2±0.6

702 Global Veterinaria, 9 (6): 700-705, 2012

Table 1: Continued Tetratoxum 37.5 1.7±2.5 50 2.7±3.9 50 2.5±3.2 Unifasciculatum 12.5 0.8±2.2 25 1.7±3.8 50 2.1±2.6 Parvum 25 0.9±1.8 25 1.0±2.2 12.5 0.4±1.2 Spirodinium 37.5 1.0±1.6 12.5 0.1±0.3 - - Equi 37.5 1.0±1.6 12.5 0.1±0.3 - - Triadinium 50 3.5±3.8 75 6.9±6.1 62.5 5.4±5.3 Caudatum 50 3.5±3.8 75 6.9±6.1 62.5 5.4±5.3 Gassovskiella 12.5 0.2±0.6 - - - - galea 12.5 0.2±0.6 - - - - Allantosomatidae Allantosoma 12.5 0.6±1.7 37.5 3.5±6.4 - - Intestinale 12.5 0.6±1.7 37.5 3.5±6.4 - - Total Genera 18 16 16 Species 28 27 20

Table 2: Number of ciliate species in the feces per animal of Equids Mean SD Range Horses 12.8 3.0 8.0-18.0 Mules 8.1 5.8 0-16.0 Donkeys 7.5 4.0 0-12.0

Table 3: Number of ciliates (ciliate density) in the feces of Equids Mean* SD** Range Horses 15.4 10.8 5.5-37.5 Mules 6.4 4.6 0-12.5 Donkeys 10.4 13.2 0-39.0 *Mean number of ciliates (x1041 cells ml ) **Standard deviation

Table 4: Total ciliate abundance and distribution of the total number of genera and species of ciliates from the hindgut contents of equids from various locations around the world Localitya Total ciliates (x1041 cells ml ) Range (x1041 cells ml ) Total no of genera Total no of species References China c c 19 30 22, 32, 33 Japan 3.4cc 19 40 9 Japan 9.0c 0.4-113.0 22 49 10 Taiwan 38.1±35.9b 0.3-127.0 19 38 12 Japan 1.4c c 11 18 13 Japan 140.0c c 23 50 14 Middle Asia c c 25 57 15 Cyprus 3.0±2.5b 0.5-8.5 16 22 17 Turkey 14.2±13.9b 0-45.5 22 36 18 Cyprus 5.5±4.4c 1.5-12.5 9 11 20 Turkey 26.4±15.1b 0-54.5 21 37 19 Turkey 15.4±10.8 5.5-37.5 18 28 Present study Turkey 6.4±4.6 0-12.5 16 27 Present study Turkey 10.4±13.2 0-39.0 16 20 Present study aNumber of animals and breed, listed in descending order from the top of the Table. China: 20 horses, donkeys and mules; Japan: 17 light horses; Japan: 60 race horses; Taiwan: 40 riding horses; Japan: 20 Tokara ponies; Japan: 18 Kiso horses; Middle Asia: 184 kulans; Cyprus: 13 wild donkeys; Turkey: 15 Rahvan horses; Cyprus: 5 domestic horses; Turkey: 15 racing horses; Turkey: 8 domestic horses; Turkey: 8 mules; Turkey: 8 donkeys. bMean±SD cData not reported

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