European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

First Record of spp. from sheep in Basrah City, Southen Iraq Israa Mohsen Essa 1, Ghazi Yaqoub Azzal 2

1, 2 Department of Microbiology, College of Veterinary Medicine, University of Basrah, Basra, Iraq Abstract Trichuris spp. were isolated from slaughtered sheep in the South Regions of Iraq / Basrah. The diagnosis and description were based on morphological characteristics. Two of Тrichuris were found, T. ovis and T. globulosa. While T.оvis was most common, Т. globulosa was relatively rare among the infested . Male and female nematodes were recognized as Trichuris spp. morphologically; however, only one male was identified as T. globulosa. The species of Trichuris spp. were distinguished through their biometrical features such as presence of cylindrical spicular sheath, protrusive vulva, length of the spicule, as well as the proximal and distal cloacal tube. Moreover, diagnosis of Trichuris species was confirmed by the nomenclatural acts as it has been recorded in the registration system of the Iraqi Natural History Research Center when a certain deposit number for Trichuris spp.: INHM.2020Nem.1.1. They were noting the presence of Trichuris spp. as such the samples of nematodes were deposited in the helminth collection at the Museum of the University of Baghdad. This repository is considered as the confirmation of the diagnosis and morphological analysis.

Keywords: , Trichuris globulosa, Morphological characters, Sheep, Basra.

INTRODUCTION Nematodes of the Trichuris (Adenophorea: ) Roederer 1761, is cosmopolitan and destroy awide variety of mammalian hosts, including [1, 2]. The Trichuris genus includes medicinal and veterinary species importance; has been acquired by domestic animals from many countries or is shared with them [3]. Trichuris ovis is acaecal parasite most prevalent in ruminants irrespective of age, gender, and breed of the hosts [4]. Trichuridae nematodes are one of the most diverse and widely distributed groups of parasitic worms. It has a clear economic impact, as it may cause the death of economic animals, including sheep, which in severe cases; suffer from bloody colitis and diphtheritic caecitis, causing ulcerative and necrotic mucosal lesions. Extreme anemia and dehydration, and jaundice in animals with high worm burden can contribute to death [5, 6, 7]. There are more than 70 species of Trichuris nematodes parasitic recorded in primate mammals such as ruminants, lagomorphs, even-toed ungulates, rodents, carnivores [8, 9].

355

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

Trichuris nematodes are common in ruminant animals in many countries, especially sheep [10, 11]. Previous reports indicate that there are three species of trichurid nematodes in Europe; T. skrjabiny, T. abildgaard, and T. globulosa [12, 13]. Also, Т.ovis was not limited to Europe, since it was found in India, Nigeria, and South America as well [14, 15]. Recently, the species Т. globulosa was recorded in Iran [16] and in Ukraina [17]. Adults of T.ovis can be recognized by their white in colour, their characteristic, slender, long thread-like anterior portion, and a much thicker posterior end. Trichuris spp. got its name due to its thin, thread-like anterior body end, but the most common name is ovine whip worm [4]. Regarding T.ovis, life-cycle is direct; the transmission can be by the soil. The embryonic eggs could orally be ingested, and then hatched into the small intestines. The parasite larvae at the intestinal wall of the caecum and proximal colon and develop into mature worms [18, 19]. There is limited information about the T.ovis adult worm and its infection in the sheep caecum in Iraq. In Basrah, anothers pecies of nematode infection of gastrointestinal tract was studied in sheep [20]. No previous researches recorded T. ovis as an adult worm in Iraq, which leads to the limited information about the worm and its infection methods in the sheep caecum in Iraq. Therefore, the present study mainly aimed to isolate and diagnose Trichuris species in slaughtered sheep in Basrah province, Iraq.

MATERIALS AND METHODS Samples collection Three hundred and sixty caecum samples were collected randomly from the central slaughterhouses in different areas in Basrah province including Karmat Ali, Al-Jumhuriya, Al- Mawfakia, Al-Abasiya, Al-Jubaylah, and Al-Hakimiah during 2019 to 2020. Complete information was collected about the important sources and breeding location of the animals (sheep), which help in determining the origin of the infection. The samples were brought to the parasite research laboratory of the University Basrah/ Veterinary Medicine.

Laboratory study Examination and collection of parasites The cecum was incised longitudinally by using special surgical tools. The presence of parasitic worms is investigated either by examining the fecas in the cecum or in the cecal epithelium to which the worm attaches. Helminths were obtained through the use of modified techniques [18, 21]. Nematodes were isolated from cecum in the laboratory, the cecum contents emptied into a bucket, the cecum mucosa was washed separately in clean water, and all the parasites were kept, the isolated worms were washed several times with distilled water and then transferred to sterile plastic containers, and These worms were preserved in 70% alcohol, were dried in glycerol alcohol (10 parts glycerol and 90 parts 70% alcohol) [14]. Then the preserved nematodes were processed on glass slides in anhydrous glycerol. Slides were observed under a light optical microscope, and images were taken under a search microscope at ×4, ×10, and ×40. Fixation and preservation Nematodes were fixed immediately after collection for 24 hours in 70% alcohol. A mixture of 70% ethyl alcohol as well as the glycerol was used to streamline the parasites. Gender species

356

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021 females and males were described, and nematodes were microscopically recognized by the standard taxonomic keys [5, 7, 22]. The morphology of the spicules was the key characters used for the identification of male nematodes and photographed where appropriate. After identification, the following measurements were made for many worms: body length, spicule length, esophageal length; these features were measured using an eyepiece graticule mounted in 10× eyepiece and 40× objectives, calibrated initially using a stage micrometer and presented as the range (measurements micrometers) [7]. Taxonomic identification based on morphological studies of Trichuris spp. The phenotypic characteristics of isolated worms (length and colour) were recorded. The worm length was measured with a standard ruler; Trichuris Nematode males were selected from all examination caecum; lactophenol was used to clear the spicules under a microscope in the male worms, as described by [23]. Each worm was examined for morphometrical spicule parameters. The sample was sent for registration and confirmation of diagnosis and classifications. This Nematoda and the nomenclatural acts it registered in the registration system of the Iraqi Natural History Research Center.

RESULTS Trichuris spp. genus nematodes were found to be a spread parasite that infects sheep in Basra/ Iraq. The intensity of the parasite presence was from 1 to 30 parasites per cecum. The species of Т. ovis and Т. globulosa were found in most positively infected sheep. The species of Т. globulosa was rarely detected. The morphological characteristics of the adult species Trichuris spp. showed that there was clearly the creamy white colour and their slender filiform, anterior end comprising about three-fourths of the complete body length this study (Figures 1, 2).

♂ ♀

Figure (1): Adult Trichuris spp. attached Figure (2): Adult Trichuris spp. with mucosa cecum organ inpetridish with normal saline

Collected Trichuris genus nematodes have anterior thin, long, translucent parts, rather than a thicker, short, and usually white posterior part (Figures 3, 4).

357

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

♀ ♂

Figure (3): Male of Trichuris spp. on Figure (4): Female of Trichuris spp. with slide ruler measurement

The males have a slightly spiral tail end (Figure 3); whereas, the tail end of females was slightly curved (Figure 4). Females are significantly longer than males. The ratio of anterior to posterior body parts varied significantly (3: 1) in females and (2.8: 1) in males. The female body at the posterior end was significantly wider than male bodies. The characteristics of morphological and metric species (body (size, form and structure) were considered as typical differential in both adult males and females sexual morphism is very distinct in body size, the sexual apparatus, and tail-end structure (Тable 1).

Тable (1): Morphometric parameters of Т. оvis and Т. globulosa from sheep Characters Female Male T. ovis T. ovis T. globulosa Length of body (mm) 57.77 ± 1.51 49.73 ± 1.35 48.83 ± 1.03 Anterior to posterior body parts ratio 3.0 : 1 2.2 : 1 2.9 : 1 Width of body at the anterior end (μm) 198.52 ± 6.19 186.98 ± 5.60 146.92 ± 2.38 Width of body at the posterior end (μm) 912.06 ± 17.42 720.90 ± 7.37 667.44±7.42

The anterior body part consists of a small round mouth with thin lips (Figure 5A), followed bythe esophagus, which is recognized by long, occupying (Figure 5B). The specific morphology of the Trichuris genus, the structure cuticle surface, is slightly gentle transverse striations (Figure 6). The adult females' have the genital tract connected by a vulvar opening, which Showing an oviduct with eggs as a single profile in and bearing fully developed eggs (Figure 7). Recognized of the vagina T.ovis was long, muscular, and curved. The vulva itself extrudes as two lips of slightly posteriorly bent cylindrical cuticular protrusion. This protrusion is covered with numerous spines (Figure 8A). In most of T. ovis females, eggs were arranged in several rows (Figure 8C), or accumulated in the middle or distal part of the vagina (Figure 8B).

358

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

A a

B b

Figure (5): Anterior end of Figure (6): Cuticle surface of Trichuris spp. Trichuris spp. (100×) (100×)

B b a

A

C

c

Figure (7): T. ovis eggssingle profile Figure (8): T. ovis showing egg exeste in (100×) vulvar opening (100×).

Eggs of Trichuris spp. are lemon-shaped with a plug at each of the poles, Measuring are (70.4- 82.3×34.5-45.4μm as length and width respectively (Figure 9). The posterior endofthe female body was significantly wider than male bodies.The posterior extremity was slightly curved (Figure 10).

359

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

Figure (9): Egg of Trichuris spp. (100×) Figure (10): Posterior end of female Trichuris ovis (×400)

The adult males of T.ovis can be distinguished by the posterior end, characterized by the slightly spiral bent tail and the spicule was long with a specular sheath covered with spines and which spirally coiled with a single spicule, having expanded proximal and a pointed distal end (Figure 11). While in T. globulosa, it is theplainly rounded tail, the spicular sheath had a slightly stretched globular expansion at its distal end and was covered with closely set spines (Figure 12). To identify T. ovis and T. globulosa, we took into account the following morphological features; in males, the structure of the tail end, namely the spicule and the spicule sheath.

Figure (11): Tail end of male Т. оvis (×100) Figure (12): Tail end of male Т. globulosa (×100)

360

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

The parts of tail end male Т. оvis were shown, which consists of (A) the distal end of spicule, (B) spicule, and (C) proximal end of spicule (Figure 13). The parts of tail end male Т. globulosa were consists of (A) distal end of the spicule, (B) spherical dilation for a distal end of spicule sheath, (C) proximal end of the spicule, (D) spicule, (E) spines at spicule sheath, and (F) cylindrical protrusion at the apex of the spherical dilation with spicule sheath (Figure 14).

B

C D F A

B A

E C

Figure (13): Male of Т. globulosa (×100) Figure (14): Male of Т. globulosa (×100)

This work and the nomenclatural acts it contains have been registered in the registration system of the Iraqi Natural History Research Center: Trichuridae: (Ablidg, 1795, INHM: 2020 Nem. 1.1), and the Museum / University of Baghdad, samples of nematodes have also been deposited in the helminth collection of the same museum where it is considered to confirm the diagnosis and Morphological analysis. Through our current study we found that one more character- specific of T. globulosa males. It was not previously described in the literature, and, in our opinion, can facilitate its identification. It is the cylindrical protrusion at the apex of the spherical dilation of the withdrawn spicule sheath (Figure 15). This character is absent in T. ovis males that are morphologically very similar to those of T. globulosa. The length of the cylindrical protrusion was 81.43 ± 2.67μm, and its width was 61.59 ± 0.45μm. Summarizing the results, it can be noted that nematodes of the genus Trichuris are common in domestic sheep in the southe regions of Iraq Trichuris nematodes of sheep here are represented by two species, T. ovis and T. globulosa, which T. ovis is more prevalent.

A B

361

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

C D

E F

Figure (15): Drawings of Trichuris ovis. (A): In esophagus, muscular and stichosome portions; (B): In female, detail of vulva, and bacillary band (lateral view); (C): Egg; (D): Esophagus- intestine junction; (E): In male, posterior end, spiny spicular sheath, spicule, and proximal and distal cloacal tube (lateral view); (F): In female, posterior end (lateral view)

DISCUSSION Previously, the morphometric characters of many Trichuris species were analyzed [17, 24]. Produced key to seven whipworm species from ruminants in North America were including T. ovis and T. skrjabini [25]. During the present study, was distinguished two species of Trichuris as T. ovis and T.globulsa was distingushed isolated from South Iraqi sheep cecum, as it detected that the main diagnostic characters are body length, spicule length and width, spicule shaft diameter and the length of the ejaculatory duct [25, 26]. According to previous studies of various authors, sheep commonly host four Trichuris species: T. ovis. T. globulosa, T. skrjabini and T. capreoli [27, 28]. However, doubts have arisen regarding the relevance of T. globulosa. On the basis of morphological characteristics, it is shown that discovered the presence of only T. globulosa males and T. ovis females in the same host [29]. The absence of T. ovis males and T. globulosa females in the same host suggested sexual reproduction between both species, and thus, they represent only one species. Therefore, these authors believe that T. ovis and T. globulosa are synonymous. This is what our current study agreed upon T. globulosa in my

362

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021 study was observed in the same host present the presence of only T. globulosa males and T. ovis male and females. While in this study, ahigher and more pronounced with details drawings of Trichuris globulosa was done,as will as different the general morphological pattern in the male reproductive system. Identification of closely related species is challenging. This is due in part to the phenotypic plasticity of the organisms themselves, host-induced variation, the lack of morphological features, and the extensive overlap in morphometric characteristics among species [26]. Moreover, some identical isoenzyme patterns were found through isoenzyme electrophoretic studies [29]. Despite the fact that Trichuris nematodes are common endoparasites in wild ruminants, especially in roe deer, much less attention is given to species found in these hosts in comparison to those found in farm animals. All-female nematodes in this study were morphologically identified as T. ovis, while only one male was identified as T. globulosa. Rickard and Bishop were suggested the following characters as suitable for species determination (male specimens): short spicule with rounded tips and large caudal papillae and long ejaculatory duct [25]. The most recent review of T. discolor can be found. T. discolor been reported worldwide, especially in cattle [24]. Findings of T. discolor in wild sheep are reported rarely [30, 31]. Furthermore, T. discolor occurs as well in the caecum and colon of ox, zebra, buffalo and goat [32]. Our study provides the first morphological analysis of Trichuris nematodes found in sheep of Basrah [12]. In the past, especially in the last century and earlier, the different nematode species were described solely by morphological characters. Morphological details of T. ovis and diagnostic structures although the bacillary band can be differentiated from the remaining transversal striated cuticle were examined using light microscopy [33, 34]. Since the bacillary bands are not known for all Trichuris species, it is possible that the real importance of this structure will be assessed only when new studies with detailed descriptions, illustrations, and photographs are provided. Trichuris ovis and T. globulosa has been differentiated by based on the size of the spines on the distal expansion of the everted spicule sheath (those on the distal expansion of the sheath being larger than the rest in T. globulosa and vice versa in T. ovis) However, no size difference was observed in the spines of the spicule sheath in the male specimens of T. ovis examined herein [35]. The males of the two species could be differentiated with reference to the shape of the proximal end of the spicules and also of the distal expansion of the everted spicule sheath. Further, no cuticular inflation was noticed near the head end, as recorded by Farleigh [36]. From the existing species of genus Trichuris Roederer, 1761, the present specimens were found to be very close to the description of Trichuris ovis as it is steated [37]. As regards its morphological characters, including the total length, maximum breadth, size of the spicule, anterior end, etc. However, some intraspecific variations in size ratio have been recorded in the present specimens, which are of minor taxonomic. However, most authors indicated that T. ovis are the most prevalent species, explaining it with the faster development cycle of this parasite [15, 38]. In our opinion, the local dominance of T. ovis can be explained by favourable environmental conditions for the development of embryonic stages of parasites. The morphological similarity of T. globulosa and T. ovis males, and also the fact that it is not always possible to isolate males of one species inspired us to obtain new data on the significantly different morphometric characteristics of males and females of studied Trichurus species. We established that, in addition to sex-related characters, it is necessary to

363

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

take into account the morphological features of the metric indices of cuticle formations on their body surface. It is clear from the study that ruminants of Iraq make no exception from the rest of the world regarding Trichuris ovis infection. Therefore it is believed that the present study will be of some use to avoid this confusion regarding the identification of this parasite.

REFERENCES [1] Cafrune, M.M.; Aguirre, D.H.; Rickard, L.G., (1999). Recovery of Trichuris tenuis Chandler, 1930, from camelids (Lama glama and Vicugna vicugna) in Argentina. J. Parasitol. 85, 961–962. [2] Anderson, R.C., (2000). Nematode Parasites of Vertebrates. Their Development and Transmission, 2nd ed. CAB International ed, Wallingford Oxon UK. [3] Mirdha, B.R.; Singh, Y.G.; Samantray, J.C.; Mishra, B.; (1998). infection in slum children. Indian J. Gastroenterol. 17, 154. [4] Iliev, P.T., et al. "Some parasitological, pathological and immunohistochemical examinations in sheep naturally infected by Trichuris ovis." Trakia Journal of Sciences 2 (2017): 174- 178.‏ [5] Soulsby, E. J. L. (1982). Helminths, arthropods and protozoa of domestic animals. 7th Ed. Baillière Tindall. London. Pp:133-250. [6] Bowman, D. D.; Lynn, R. C. and Eberhard, M. L. (2002). Helminthes. In: Saunders, W. (Ed.), Georgis’ Parasitology for the Veterinarians. Harcourt Health Sciences Company, New York. Pp:162-380.\ [7] Taylor, M. A., Coop, R. L., and Wall, R. L. (2016). Veterinary Parasitology Chichester. West Sussex, UK: Willey-Blackwell. [8] Callejón, R., Cutillas, C., Nadler, S. A. (2015). Nuclear and mitochondrial genes for inferring Trichuris phylogeny. Parasitology Research, 114 (12), 4591–4599. doi: 10.1007/s00436- 015-4705-7. [9] Hillman, A. E.; Ash, A. L.; Kristancic, A. R.; Elliot, A. D.; Lymbery, A. J.; Robertson, I. D.; Thompson, R. C. (2017). Validation of various parasite detection tests for use in the Australian marsupials quenda (Isoodon obesulus) and brushtail possums (Trichosurus vulpecula). Journal of Veterinary Diagnostic Investigation. Journal of Veterinary Diagnostic Investigation, 29 (1), 64–75. [10] Gul, N.; Tak, H. (2016). Prevalence of Trichuris spp. in small ruminants slaughtered in Srinagar District (JandK). Journal of Parasitic Diseases, 40 (3), 741–744. [11] Vejl, P.; Nechybová, S.; Peřinková, P.; Melounová, M.; Sedláková, V.; Vašek, J.; Čílová, D.; Rylková, K.; Jankovská, I.; Vadlejch, J.; Langrová, I. (2017). Reliable molecular differentiation of Trichuris ovis and from sheep (Ovis orientalis aries) and roe deer (Capreolus capreolus) and morphological characterization of their females: morphology does not work sufficiently. Parasitology Research, 116 (8), 2199– 2210. [12] Oliveros, R.; Cutillas, В. C.; de Rojas, M.; Arias, P. (2000). Characterization of four species of Trichuris (Nematoda: Enoplida) by their second internal transcribed spacer ribosomal DNA sequence. Parasitology Research, 86, 1008–1013. [13] Cutillas, C., Oliveros, R., de Rojas, M, Guevara, D. C. (2004). Determination of Trichuris skrjabini by sequencing of the TS1–5.8S–ITS2 segment of the ribosomal DNA:

364

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

comparative molecular study of diff erent species of trichurids. Journal of Parasitology, 90, 648–652. [14] Kuchai, J. A.; Chishti, M. Z.; Zaki, M. M.; Ahmad, J.; Rasool, M.; Dar, S. A.; Tak, H. (2011). Prevalence of nematode parasites in Sheep of Ladakh-India. Journal of Agricultural Extension and Rural Development, 3 (13), 229–231. [15] Jegede, O. C.; Adejoh, A. A.; Obeta, S. S.; Olayemi, O. D. (2015). Gastrointestinal Parasites of Sheep and Goats in Gwagwalada Area Council, Federal Capital Territory, Abuja, Nigeria; with a special reference to sex, breed and age. Alexandria Journal of Veterinary Science, 46 (1), 170–176. [16] Bahrami, A. M., Hosseini, E., Baran, A. I. 2016. A study on histopathological changes due to zoonotic nematodes in sheep in Ilam province, Iran. Journal of Zoonotic Diseases, 1 (1), 47–53. [17] Yevstafieva, V. A.; Yuskiv, I. D.; Melnychuk, V. V.; Yasnolob, I. O.; Kovalenko, V. A.; and Horb, K. O. (2018). Nematodes of the genus Trichuris (Nematoda, Trichuridae), parasitizing sheep in central and South-Eastern regions of Ukraine. Vestnik zoologii, 52(3), 193-204.‏ [18] Soulsby, E. J. L. (1986). Text book of: helminths, arthropods, and protozoa of domesticated animals. [19] Zainab, T., and Khan, W. (2016). Morphological and histopathological studies of Trichuris ovis in goat intestine. J. Parasit. Dis. Diag. Ther, 1, 122-129.‏ [20] AL-Bahi, A. (2011). Some epidemiological and diagnostic aspects study on abomasal nematodes in the sheep of Basrah province\ Iraq.M.Sc Thesis, Col.Vet. Med., Basrah. [21] Hansen, J. and Perry, B. (1994). The epidemiology, diagnosis and control of helminth parasites of ruminants. International laboratory for Reserch on Animal Diseases. [22] Gibbons, L. M. (2009). Keys to the nematode parasites of vertebrates. Supplementary volum. Cambridge University Press, Cambridge. Pp:352. [23] Jacquiet, P .; Cabaret, J. ; Cheikh, D. and Thiam, E. (1997) . Identification of Haemonchus Species in Domestic Ruminant based on Morphometrics of Spicules. Parasitol. 83:82-86. [24] Callejón, R.; Halajian, A.; De Rojas, M.; Marrugal, A.; Guevara, D.C.; Cutillas, C., (2012). 16S partial gene DNA and internal transcribed spacers ribosomal DNA as differential markers ofTrichuris discolorpopulations. Vet. Parasitol. 186, 350–363. [25] Rickard, L.; Bishop, J.; (1991). Redescription of Trichuris tenuis, Chandler, 1930, from llamas (Lama glama) in Oregon with a key to the species of Trichuris present in North American ruminants. J. Parasitol. 77, 70–75. [26] Robles, M. D. R. (2011). New species of Trichuris (Nematoda: Trichuridae) from Akodon montensis Thomas, 1913, of the Paranaense forest in Argentina. The Journal of parasitology, 97(2), 319-327. [27] Kuzmina TA, Kharchenko VA, Malega AM (2010) Helminth fauna of Morgan, B. and P. A. Hawkins. (1960). Veterinary helminthology. 5th Ed., Burgess Publishing Company, U.S.A [28] Aukštikalniené R, Bukelskis E, Kašetaité E (2007) Intestinal helminthes of Cervidae in the Aukštaitija National Park. Balt For 13:96–102. [29] Cutillas, C.; German, P.; Arias, P.; Guevara, D. C. (1995). Trichuris ovis and Trichuris globulosa: morphological, biometrical and genetic studies. Experimental Parasitology, 81, 621–625.

365

European Journal of Molecular & Clinical Medicine

ISSN 2515-8260 Volume 08, Issue 02, 2021

[30] Allen, R.W. (1955) Parasites of mountain sheep in New Mexico, with new host records. J Parasitol 41:583–587. [31] Blood DA (1963) Parasites from California bighorn sheep in southern British Columbia. Can J Zool 41:913–918. [32] Cordero del Campillo M, Rojo Vázquez FA (1999) Parasicología Veterinaria. McGraw-Hill, Interamericana, Madrid. [33] Rossin, M.A, and Malize, A.I. (2005). Redescription of Trichuris pampeana (Nematoda: Trichuridae) from the South American subterranean rodent Ctenomys talarum Thomas, 1898 (Rodentia: Octodontidae). Journal of Parasitology, 91 (1), 127-130. [34] Robles, M.d.R.; Navone, G.T.; Notarnicola, J.; (2006). A new species ofTrichuris (Nematoda: Trichuriidae) from Phyllotini rodents in Argentina. J. Parasitol.92, 100–104. [35] Baylis HA. Note on the cestode Moniezia (Furhmanella) Transvoalensis (Baer, 1925). Mem Ind Mus 1935; 9(8): 248-51. [36] Farleigh EA. Observation on the pathogenic effects of Trichuris ovis in sheep under drought conditions. Aust Vet J 1966; 42(12): 462-63. [37] Schimdt GD. (1986). Hand book of Tapeworm Identification. CRC Press, Inc. Boca Raton, Florida 1986; p. 675. [38] Kuchai, J.; Ahmad, F.; Chishti, M.; Dar, J.; Tak, H.; (2013). On Morphology and Morphometry of Trichuris ovis Abildgaard, 1795 recovered from Ruminants of Ladakh, India. Journal of Buffalo Science, 2:49-52.

366