Prevalence of gastro-intestinal parasites in horses ( Equus caballus linnaeus , 1758) in Setif region (north-eastern of Algeria)

Abstract Background Horses are used in various countries for multiple purposes and often play a crucial role in the livelihood of many communities. However, parasitic infections represent a significant threat to their health and productivity.Objectives: This study aimed to identify gastrointestinal parasite species affecting horses using coprological methods and to assess the influence of certain factors (age, sex, and bioclimatic zone) on these infections.Study design: These samples were analyzed using flotation techniques to detect fecal parasitic forms (eggs, oocysts) and larvae.Methods: Between January and May 2024, fresh fecal samples were collected from 121 horses in Setif region (Northeastern Algeria). Prevalences were calculated for each gastrointestinal parasite and the influence of certain intrinsic (age, sex) and extrinsic (region) parameters on these prevalences was analyzed. We have found, namelyStrongylus vulgaris (38.84%), Oxyrus equi (7. 44%), Parascaris equorum (24.62%), Anoplocephala spp. 6.61% and 5.79 were carriers of Eimeria leuckarti and 4.96% had Giardia duodenalis cysts

This study demonstrates that horses in the Setif region are commonly affected by gastrointestinal parasites, with a predominance of mixed infestations. Both helminths and protozoa were identified, withStrongylus vulgaris and Parascaris equorum being the most prevalent species. The variations in infestation rates across age groups, sex, and geographical zones highlight the influence of environmental and management factors.Keywords : Horse, parasite, gastrointestinal, Algeria Title : Prevalence of gastro-intestinal parasites in horses (Equus caballus linnaeus , 1758) in Setif region (north-eastern of Algeria) Authors : Badredine Bouchama(1); Hamza Khaled(2) ; Achour Mennani(3) 1. Laboratory of Improvement and development of plant and animal production, University of Setif 1 Ferhat Abbas 2. Laboratory of Animal Biotechnologies, Institute of Veterinary Sciences, University of Blida 1 3. Department of Agronomic Sciences, Faculty of Sciences, University of M’sila, Algeria The corresponding author: Badredine Bouchama: [email protected] Hamza Khaled: [email protected] Achour Mennani: [email protected]

Background Horses are used in various countries for multiple purposes and often play a crucial role in the livelihood of many communities. However, parasitic infections represent a significant threat to their health and productivity. 1 Posted on 30 Jul 2025 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.175387555.57673814/v1 | This is a preprint and has not been peer-reviewed. Data may be preliminary.

This study aimed to identify gastrointestinal parasite species affecting horses using coprological

and to assess the influence of certain factors (age, sex, and bioclimatic zone) on these infections. Study design:These samples were analyzed using flotation techniques to detect fecal parasitic forms (eggs, oocysts) and larvae.

Between January and May 2024, fresh fecal samples were collected from 121 horses in Setif region (Northeastern Algeria). Prevalences were calculated for each gastrointestinal parasite and the influence of certain intrinsic (age, sex) and extrinsic (region) parameters on these prevalences was analyzed. We have found, namelyStrongylus vulgaris (38.84%), Oxyrus equi(7. 44%),Parascaris equorum(24.62%), Anoplocephala spp. 6.61% and 5.79 were carriers ofEimeria leuckarti and 4.96% hadGiardia duodenaliscysts

This study demonstrates that horses in the Setif region are commonly affected by gastroin- testinal parasites, with a predominance of mixed infestations. Both helminths and protozoa were identified, with Strongylus vulgaris and Parascaris equorum being the most prevalent species. The variations in in- festation rates across age groups, sex, and geographical zones highlight the influence of environmental and management factors.

:Horse, parasite, gastrointestinal, Algeria

Horses (Equus caballus Linnaeus, 1758) have been essential companions to humans for over 32,000 years (Cain, 2022), serving historically as sources of food, means of transportation, agricultural labor, instruments of warfare, and partners in sport (Rosa et al., 2018). Algeria has a long-standing equestrian tradition, where the national equine population is estimated at around 150,000 individuals, primarily composed of Barb horses (90%), with smaller proportions of Arab-Barb, Arabian Thoroughbreds, and French Trotters (Khaled and Besbaci, 2024). Although horses are generally resilient animals, they remain susceptible to a variety of diseases, some of which can result in high mortality rates. Among the most common equine health issues are parasitic infestations/emdash.cyrboth external (e.g., ticks and lice, which may also act as vectors) and internal. Internal parasites include hemoparasites (such as those responsible for piroplasmosis) and gastrointestinal parasites(PGI) , which can lead to severe health disorders including diarrhea, emaciation, colic, anemia, hemorrhage, and even death (Scala et al., 2020). This study focuses on gastrointestinal parasites, specifically those whose eggs, oocysts, or larvae can be identified in equine feces. The research was conducted over a five-month period (January to May 2024) and involved 121 horses in Setif region, northeastern Algeria. The main objective was to evaluate the prevalence of gastrointestinal parasites in the equine population of this region.

& Methods Presentation of the study area Setif region is located in northeastern Algeria on the high plateaus separating the Tell Atlas from the Saharan Atlas, spans an area of 6549,64 km² at an altitude of approximately 1100 meters above sea level. It borders B´ eja¨ ıa and Jijel to the north, M’Sila and Batna to the south, Mila to the east and Borj Bou Arreridj to the west. It is a semi-arid region, with an agro-pastoral vocation, characterized by cereal cultivation. This study was conducted between January and May 2024 and included 121 horses ranging in age from 3 months to 27 years, sampled from five different locations. Site selection was based on horse availability and accessibility. Environmental and breeding conditions varied across sites, providing a more comprehensive understanding 2 Posted on 30 Jul 2025 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.175387555.57673814/v1 | This is a preprint and has not been peer-reviewed. Data may be preliminary. of parasitic distribution. The study area was divided into two zones according to meteorological data: the humid zone (Ain Arnet and Serdj Elghoul) and the meridoneal zone (Ain Oulm` ene, Ain Azel and Gudjel) (Figure 1). Sampling All horse owners provided informed consent and were briefed on the objectives of the study. Horses were clinically examined and owners were questioned about grooming habits, stable and paddock hygiene, general health problems and previous anthelmintic treatments. Fresh faecal samples were collected with gloved hands in wrapped containers, labelled with the animal’s name, age, sex and date of collection, and transported thereafter to the Parasitology Laboratory. Age was classified into three categories: young (0-2) years (n=37), adult (3-15) years (n=71) and older horses (>15 years) (n=13). Laboratory examination The faecal samples were preserved in a 2.5% potassium dichromate solution (K2Cr2O7), packed in polyethy- lene bags and transported to the improvement and development of plant and animal production Laboratory (University of Setif 1 Ferhat Abbas) at room temperature (20-25°C). Coprological analysis was carried out using flotation and sedimentation concentration techniques (Zajac & Conboy, 2012) according to the follo- wing protocol: 3 g of faeces were homogenised in 42 ml of distilled water and then filtered through sterile gauze. The filtrate was then centrifuged at 1500 g for 5 minutes; the pellet was re-suspended in a saturated solution of NaCl (density 1.20) and centrifuged again at 1000 g for 3 minutes. The supernatant phase was analysed after the addition of methylene blue (0.1%) The sediment was stained with lugol (1% iodine solution) to identify helminth eggs, cysts and protozoan trophozoites. Statistical analysis Data were analyzed using the Chi-2 test based on sex, animal age, and region. SPSS version 27 was used, and the confidence level was maintained at 95%, with results considered significant at P < 0.05.

Among the 121 fecal samples analyzed, 66.1% tested positive for gastrointestinal parasites, with 64.46% of horses exhibiting mixed infestations involving more than one parasite species. Results are presented in Table 1. Coproscopic examination It was positive for different types of helminth eggs and protozoa, namelyStrongylus vulgaris (38.84%), Oxyrus equi (7. 44%), Parascaris equorum (24.62%), Anoplocephala spp.6.61% and 5.79 were carriers of Eimerialeuckart i and 4.96% hadGiardia duodenalis cysts. Figure 2 shows the prevalence of these parasites. Influence of age on PGI infestationThe data presented in Table 2 reveal that the highest overall prevalence of infestation was observed in adults, at 53.8%, followed by foals, with an infestation rate of 33.8%, and older subjects, with 12.5%. The results indicated that the age-related prevalence of intestinal parasites was highest forEmeria leuckarti at 57.1% in young animals, whileAnoplocephala spp.had the lowest prevalence at 12.5%. However,Anoplocephala spp.showed the highest prevalence at 75.0% in adults, whereasGiardia duodenalis had the lowest prevalence rate (33.3%). In older horses, the most common parasite wasGiardia duodenalis (16.7%), with Strongylus vulgaris being least common (8.5%). No cases of Eimeria leuckarti were detected in older horses. Statistical analysis showed no significant association between age and overall parasites prevalence (p > 0.05). Influence of sampling region on PGI infestation 3 Posted on 30 Jul 2025 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.175387555.57673814/v1 | This is a preprint and has not been peer-reviewed. Data may be preliminary. During our survey, we recorded 49 horses reared in the humid region and the remaining 72 in the southern region. According to Table 3, the prevalence of PGI infestation in the southern region is 63.7% and it was much higher than that of animals reared in the wet region (36.3%). Statistically, the Chi-2 test showed that there was independence between the rearing system and horses’ infestation byPGI (p>0.05), except for Emeria leuckarti (p<0.05) where a dependency between these two variables has been observed. Effect of sex on parasitic infestation According to Table 4,PGI infestation in males (72.1%) is greater than females (62.8 %), especially for Strongylus vulgaris and Giardia duodenalis (with4.6% and 4.7% respectively). In contrast, some parasitic species, such asAnoplocephala spp. (7.7%), Oxyrus equi (8.9%) andEmeria leuckarti (6.4%) prefer females as hosts than males (46.5%). At the same time, prevalence rates were similars forParascaris equorumfor the both sexes. However, no correlation was observed between the prevalence of infestation by PGI species and sex of the host (P>0.05). Except forGiardia duodenalis, where we found a correlation between infestation and sex.

Horses (Equus caballus) are frequently exposed to a variety of gastrointestinal parasites, including helminths and protozoa, which can cause significant morbidity and mortality (Hodgkinson, 2006). Our study represents the first assessment of the prevalence of these equine parasites in the region of Setif, located in north-eastern Algeria. We observed an overall prevalence of 66.1%, a result that falls within the range of values reported by several previous studies. In Ethiopia, Regassa and Yimer (2013) have found 60.68%, Yadav et al. (2014) with 63.7%, and Samuel (2015) reported similar prevalence. These results are also comparable to those of Nagwa (2023) in Ireland, with 52.40%. Compared to other regions, our prevalence was higher than that observed by Adam et al. (2013) in North Darfur, Sudan (24.6%) and by Alsahli (2024) in Riyadh, Saudi Arabia (38.93%). Conversely, it remains lower than the rates reported by Francisco et al. (2009) in Spain (89%), by Naresh Oli and Janak Raj Subedi (2017) in Nepal (84.6%), and by Tilahun (2014) in southern Ethiopia. The highest prevalence was documented by Slivinska et al. (2009) in Poland, reaching 100%. These disparities between regions could be explained by several factors, including differences in management systems, climatic and geographical conditions, sampling protocols (sample size and collection periods), diagnostic methods used (Yadav et al., 2014), and health practices implemented by equine breeders. The prevalence ofParascaris equorumobserved in this study was 25.62%. This value is comparable to those reported by Yadav et al (2014) in Ethiopia (18.75%), Tesfu et al (2014) in the same country (26.2%), and Slivinska (2006) in Ukraine (19.0%). On the other hand, it is significantly lower than the rates of 43.8% (Mezgebu et al., 2013) and 55.8% (Tilahun et al., 2014) in Ethiopia. Conversely, it exceeds the prevalences recorded in Nigeria (7.4%, Ogbein, 2022) and Riyadh, Saudi Arabia (1.76% in the study of Alsahli, 2024). This moderate prevalence ofP. equorumcould be explained by the balanced distribution of samples between adult and juvenile horses. This parasite mainly affects foals under 3 years of age, and infections are rarely observed beyond the age of 5 (Bucknell et al., 1995). According to the results presented above, the overall prevalence of pinworm disease in horses was 7.1%. This

is similar to that of Bucknell et al. (1995) with 7% in Australia, and comparable with the results of Wannas et al (2012) with 11.36%. On the other hand, these results are higher than those of Panova et al. (2024) in Russia, Ogbein (2022) in Nigeria and Sultan et al. (2014) in Ethiopia, who reported 1.4%, 4.7%, 1.31% and 1.47% respectively. However, they are lower than those reported by Kouidri et al (2020) in Algeria in their first study of equine pinworm disease, where a prevalence of 38.64% was observed. The relatively low prevalence (7.1%) ofOxyuris equi could be in relation to the sampling technique used, because Oxyuris equi eggs attach to the perianal skin, and their detection using the coprological flotation technique on fecal samples may not accurately reflect the true prevalence of the infestation. As forEimeria leucarti , it stands at 4.5%, a result close to the observations reported by Uslu and Guclu 4 Posted on 30 Jul 2025 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.175387555.57673814/v1 | This is a preprint and has not been peer-reviewed. Data may be preliminary. (2007), Bakırcı et al. (2004) in Turkey, and Alsahli et al. (2023) in Saudi Arabia, with respective prevalences of 3.7%, 5.88%, and 6.19%. In contrast, Naresh Oli and Janak Raj Subedi (2018) reported a higher prevalence (20%) in Ethiopia. We also found a significant association (p < 0.05) between parasite prevalence and geographical location of horses (wetlandvs. southern area). The observed variations in prevalence between these two regions could be mainly explained by altitudinal differences and climatic variations, which is consistent with the observations of Holland et al. (2001), higher altitude areas that are cooler and close to watercourses, had a higher prevalence than lowlands. The prevalence ofAnoplocephala spp.infestation was relatively low (6.61%), a result similar to that reported by Mirian et al. (2019) (with 6.0%). However, it was found to be higher than those observed by Belay et al. (2016), Belete et al. (2015) in Ethiopia and Carminatti et al. (2023) in Brazil, with prevalences of 3.1%, 2% and 1.92% respectively. Giardia duodenalis, a common zoonotic parasite, constitutes a public health problem affecting both humans and various animal species, including horses. In our study, the prevalence of giardiasis was found to be very low (4.96%), falling between the values reported by Mizani et al. (2025) (8.93%) and Paruch (2022) (30.43%). The low prevalence observed for these two parasites in this study could be explained by the limited exposure of horses to contaminated feed or water sources; and the rigorous application of a regular deworming protocol. The main helminths observed wereStrongylus vulgaris , which dominated in coprocultures but with a low prevalence (38.84%). Lower prevalences were obtained by molecular studies in Denmark (12.1%) (Bracken et al., 2012), Germany (1.9%) (Kaspar et al., 2016), as well as in Iran (6.5%) (Tavassoli et al., 2010) and (9.7%) (Alborzi et al., 2020). The prevalence in the present study is lower than that reported in the studies by Wosu and Udobi (2014), Adeppa et al. (2014) and Umar et al. (2013), who reported 55.3%, 52.38% and 68.8% respectively in northern Nigeria, as well as that of 81.8% (Slivinska et al., 2009) in Poland. Clinical relevance: Parasites live in the digestive tract of horses and can cause various health problems. Clinically, these parasites are significant because they can lead to: • Malnutrition , as internal parasites compete with the host for nutrients, leading to poor absorption and deficiencies in essential vitamins and minerals. • Tissue damage caused by parasite activity, resulting in inflammation, pain, and potential organ damage. • Weakening of the immune system, increasing susceptibility to infections, making the horse more vulnerable to secondary bacterial or viral infections.

This study demonstrates that horses in the Setif region are commonly affected by gastrointestinal parasites, with a predominance of mixed infestations. Both helminths and protozoa were identified, with Strongylus vulgaris and Parascaris equorum being the most prevalent species. The variations in infestation rates across age groups, sex, and geographical zones highlight the influence of environmental and management factors. The implementation of a comprehensive veterinary health program, coupled with preventive and control strategies, is essential to reduce the parasitic burden in the region. Public and private sector collaboration is critical to support horse breeders in adopting effective hygiene practices, regular deworming protocols, and improved stable management. Additional factors, such as the sharing of stalls, inadequate disinfection practices, and the reuse of rectal sleeves, may also contribute to parasite transmission. Addressing these practices through awareness cam- paigns and training can significantly enhance equine health and productivity. 5 Posted on 30 Jul 2025 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.175387555.57673814/v1 | This is a preprint and has not been peer-reviewed. Data may be preliminary. Declarations Funding information There was no financial support from any institution for the realization of the clinical cases. Authors’ declarations of interests No conflicts of interest have been declared. Ethics statement We collected fecal samples without compromising the health or welfare of the horses, therefore ethical approval was not required.

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