Epidemiological study of gastrointestinal helminths of donkeys in and around Bidar, Karnataka

Introduction

For thousands of years, the donkey has served as a beast of burden; frequently, it is a victim of savage slavery, being overburdened, under-nourished, and mistreated. Nonetheless, the donkey must rank among the most undervalued yet, significant draught animals in the world, playing a crucial role in the Third World's agricultural economy. According to estimates, animals provide 50% of the energy needed for agricultural productivity worldwide, and donkeys play a significant role in meeting this demand. The earliest domesticated donkeys appeared in Mesopotamia and Egypt around 4000 B.C. Donkeys require little care. They can therefore function on meagre foraging with minimal rest. Because of this, they are regarded as the best pack animal and the beast of burden used for hauling huge cargo through the plains, hills, and deserts in China, India, Egypt, Sudan, and Somalia. They are the most affordable and practical mode of transportation in tropical nations like India, serving the needs of washer men, potters, home builders, brick producers, and vanjara workers who still do earthwork today. There are around 40 million donkeys in underdeveloped nations, but their full economic potential is still untapped. According to the FAO's 2002 census, there are three lakh donkeys in India, making up 2.25 per cent of the total world population. There are 9000 donkeys in Karnataka and 1743 in Bidar. Hence, donkeys play a significant role in the overall economic growth of our nation, particularly in rural and hilly areas. Despite their crucial economic relevance, breeding programmes and health management have not been done in a way that will preserve and improve Indian donkey breeds. Even today, the owner and, regrettably, a lot of vets throughout the world have misconceptions about the stoic, diligent donkey. Due to their extreme poverty, donkey keepers have a narrow perspective on the health of their animals. Their ability to produce work, perform reproduction, and most of their longevity have all suffered as a result. Several bacterial viral illnesses and parasitic infestations, including glanders, strangles, ulcerative lymphangitis, tetanus, equine influenza, and herpes virus infection can affect donkeys. Babesiosis, trypanosomiasis, and gastrointestinal strongylosis are regarded as silent killers among parasitic disorders. The performance and draught ability of the donkey is hugely affected by parasitic illnesses, which are widespread and diverse. Lichtenfels (1975) [19] asserts that equids are the hosts of helminths from 28 genera and 75 species of nematodes, 2 genera International Journal of Advanced Biochemistry Research https://www.biochemjournal.com and 5 species of trematodes, and 3 genera and 24 species of cestodes. The small strongyles, commonly known as the cyathostomes (subfamily Cyathostominae within the Nematoda), make up a significant portion of the population of internal parasites in this complex group of helminths (Lichtenfels, 1975; Ogbourne, 1978; Herd, 1990) [9,28] . The primary issue noted in donkeys admitted to the veterinary facility was helminth parasite infection (Ali et al., 2001) [2] . According to Seri et al. (2004) [33] , 70.1% of donkeys in Khartoum state had nematode infections. The potentially deadly parasitic disease known as larval cyathostomosis is marked by severe diarrhoea, rapid weight loss, ventral oedema, and the presence of larvae in faeces, (Love et al., 1999; Kelly and Fogarty., 1993) [21,15] . Strongylus vulgaris causes verminous arteritis, which results in lameness, colic, and death (White et al., 1993) [45] .

According to Wheat (1975) [44] , the most frequent cause of colic is arterial lesions caused by S. vulgaris. The most typical symptom of a large strongyle infection is colic.

Although the data is insufficient, it has been estimated that thromboembolism related to S. vulgaris infection causes 90% of colic cases (Austin, 1994) [4] . Equine strongyle worms contain around 60 different species that have been discovered so far. The pattern of larval composition has changed globally, indicating increased cyathostome populations and a decline in S. vulgaris and other large strongyles (Klei, 1992) [18] . The prevalence of strongyle infection varies depending on the host's age (Uhlinger, 1993 and Linde et al., 1999) [42,20] , climate (Klei, 1992; Herd and Coles, 1995) [18] , management techniques (Linde et al., 1999) [20] , the parasite's generation period, and the infectivity of the pasture (Kelly et al., 1981) [14] .

Materials and Methods

The donkeys are close grazer that mostly looks for groundlevel grasses rather than the tops of the different bushes and trees that are present in the region. In the present study 432 donkeys irrespective of sex, age and breed were selected and based on clinical manifestations the faecal sample were collected and screened to know the prevalence of helminth infestation in donkeys. These donkeys were included from the nearby villages like Chintaki Raipally, Nagmarpally, Malgi, Gadgi, Mudhol, Yengunda and Karanji as well as donkeys belonging to individual holdings in and around Bidar.

Collection of faecal samples for Parasitological examination

A total of 432 faecal samples were collected. Freshly voided faecal samples were taken from donkeys in and around the bidar district. These faecal samples were collected in small polyethene bags or zip-lock polyethene bags, tagged with rubber bands, stored in a cold environment, and then brought to the laboratory for further analysis to determine the incidence of helminth infestation in donkeys.

Examination of faecal samples by following methods -Direct method and indirect methods.

Direct Method of faecal examination

On a glass slide, taken a little quantity of faecal samples and added a few drops of distilled water and made an emulsion. Put a cover slip on top of the mixture, which shouldn't be opaque, and inspect it with a low-power objective microscope. Search thoroughly over the whole smear for protozoan structures, worm larvae, or parasite eggs.

Indirect method of faecal examination

The faecal samples were screened using the slandered parasitological techniques (Soulsby 1982) [38] viz.

Sedimentation technique, and Floatation technique, in

Sedimentation method 1 to 2 gm of faecal pellets were emulsified in 10-15 ml of distilled water using a mortar and pestle. After being filtered through a sieve to remove all the large particles and debris, the emulsion was centrifuged at 2000 rpm for 3 minutes. A drop of sediment was put on a clean glass slide, combined with a few drops of distilled water and then covered with a cover slip after the supernatant fluid had been decanted. After that, the silt was inspected with low-power objective (10x) microscope lenses to detect the presence of helminth eggs.

In Floatation technique 1 to 2 grams of faeces thoroughly mixed with distilled water using a mortar and pestle. A vertical flotation tube with a flat bottom measuring 7.5 cm by 2.5 cm and a depth of one-third was filled with this mixture. After that, a saturated sodium chloride solution was added to the tube, being careful not to let the contents spill out. The tube was constantly covered with a cover slip so that it was in touch with the fluid. For all the eggs to float up, the tube was left to stand for 30 minutes. The cover slip was swiftly placed on a glass slide, which was then quickly viewed with a low-power objective (10x) for the presence of helminth eggs.

A quantitative faecal investigation (McMaster egg counting technique) was carried out to determine the level of strongyle infestation in the known weight of the faecal sample. The severity of the sickness was determined by counting the number of eggs per gram of donkey faeces. Modified Gordon and Whitlock technique -Two grams of faeces was weighed and taken in a mortar and pestle and then with the measuring cylinder, 60ml of the saturated salt solution was taken, mixed with the faecal sample and passed through the sieve to form a suspension in the beaker, and further using a pipette the above-formed faecal suspension was stirred and charged in both the chambers of McMaster's slide.

EPG = Total number of eggs counted in both chambers × 100

Statistical analysis:

The data obtained in the research work undertaken were statistically analysed and compared as per the standard statistical procedure suggested by Sendecor and Cochran (1994) [36] and the significance of the mean difference was tested by ANOVA test.

Results

In the current study, helminth infestation was assessed in 432 donkeys of diverse sexes, and ages. Moreover, donkeys with helminth infestations were observed to have clinical symptoms. A total of 18 strongyle-positive animals were selected for drug trials. Six donkeys were selected as controls for further haematological and biochemical analysis. These donkeys looked to be healthy and had no evidence of parasite infestations.

Prevalence

Age wise prevalence of Helminth infestation

The age-specific prevalence of helminth infestation in donkeys given in table-3, depicted in figure-3. According to the study's findings, donkeys aged 0 to 5 years had the higher age-specific prevalence of helminth infestation (75.48%), followed by those aged 6 to 10 years (66.67%), while donkeys older than 10 years (53.08%) had the lowest prevalence.

Sex-wise prevalence

The Sex-wise prevalence of helminth infestation in donkeys is shown in table-4, depicted in figure -4, study reveals that females (66.52%) had a higher prevalence than males (64.73%).

Season-wise prevalence

The seasonal incidence of helminth infestation in donkeys is listed in table-5, depicted in figure-5. The current study findings showed that the prevalence of helminth infestation was lowest in donkeys during the winter season (52.50%) and that it was highest in the rainy season (73.52%), followed by the summer season (67.60%).

Prevalence of Helminth infection in donkeys by direct method and indirect method

The observations on the diagnosis of faecal examination by the direct method are presented in table-7. Out of 432 faecal samples, 260 (60.18%) were found to be helminth-infested by the direct method of faecal examination and 284 (65.74%) samples were found to be helminth-infested by the indirect method of faecal examination samples.

Discussion

Prevalence Donkey faeces from 432 different animals were analysed for helminth infestation. 284 (65.74%) of these individuals tested positive for helminth infestation. In the present study, the overall prevalence of helminth infestation in donkeys is (65.74%), the finding of the present study is almost similar to the findings of Tudor (2010) [41] (68.48%), Molla et al.

(2015) (64.61%), and Alshaibani et al. (2019) [3] (67.75%). However overall prevalence was differ from that reported by Khan et al. (2010) [17] (31.0%), Imam et al. (2008) [13] (37.48%) and Bhavanakumar and Satichidanadam (1987) (22.0%), Seri et al. (2004) [33] and Ayele et al. (2006) [5] . In the present study, the mixed helminthic infestation was recorded in 50 (17.60%) cases of donkeys. The present finding of mixed helminthic infestation prevalence is in agreement with the finding of Shadia et al. (2018) [34] (24.2%), Khajuria et al. (2004) [16] (20.84%), and Ibrahim et al. (2011) [12] (35.40%). However, the present finding of mixed helminthic infestation prevalence of this study differed from others Taye (2017) [40] (10.4%), Mahfooz et al.

(2008) [22] (5.00%), and Waqas et al. (2014) [43] (5.0%) in comparison to previous studies, the current analysis found a higher incidence of helminth infestation in the study area might be associated with the lack of any parasitic helminthes intervention programme and the management systems in the area, where many donkeys were allowed to graze together on small plots of land throughout the year, which facilitates contamination between animals. (Parsani, 2008) [30] . There were Strongyles eggs in 147 (51.76%) of the positive faecal samples in the present study. These findings were in agreement with Parsani et al. (2013) [30] , Mahfooz et al. (2008) [22] and Negash et al. (2021) who reported the strongyle positive in donkeys at 55.3%, 50% and 53% respectively. Contrary to these findings were reported by Esmaeel (2010) [8] (70.0%), Chitra et al. (2011) [2] (76.12%), Mezgebu et al. (2013) [27] (66.67%), and Takele and Nibret (2013) [39] (65.09%), the prevalence of Strongyle was higher. In contrast to earlier research, the current study had a low prevalence of Strongyles spp. (51.76%). The reason for the low prevalence is thought to be related to the adverse environmental conditions for the survival of the extrinsic stages of the parasite, the inability of strongyles to undergo inhibition inside the host, and long generation intervals.

Age wise prevalence

According to an analysis of the clinical study's data, donkeys aged 0 to 5 years had the highest frequency of helminth infestation (75.48%), followed by those aged 6 to 10 years (66.67%), while donkeys older than 10 years had the lowest incidence (53.08%). The findings of this study concur with those of the articles by Ajabe et al. (2020), Mathewos et al. (2021) [23] , and Nagar et al. (2022) [25] . The delayed development of substantial immunity, which is initially low but rises with the intensity and duration of infection exposure, has been blamed for the high incidence of infection seen in the 0-5 year age range (Sabir and Bandyopadhyay 2016) [32] . Adults, however, have a reduced prevalence, which has been linked to the development of parasite immunity as a result of repeated exposure (Shah-Fischer and Say, 1989) [35] .

Sex and season wise prevalence

According to the Sexwise prevalence survey, female donkeys were more likely than males to have helminth infestations. The results of this investigation were in agreement with those of Mathewos et al. (2021) [23] , Nagar et al. (2022) [25] , and Hamed et al. (2019) [9] . Due to their physiological characteristics, which frequently result in stress factors and lower their immunity to infections, female animals who are breastfeeding parents are weak and malnourished, making them more vulnerable to illnesses (Radostits et al., 2007) [31] . Helminth infection was shown to occur more frequently in donkeys during the rainy season (73.52%), followed by the summer season (67.60%) and the winter season (52.5%). The results of this investigation were consistent with those of Nagar et al. (2022) [25] , which may be because nematode larvae are more readily available in pastures during the wet season (July-September). In addition to lowering host immunity, rain and high relative humidity during the rainy season favour ecdysis and larvae dispersal, increasing the likelihood of infection (Soulsby, 1966) [37] .

Conclusion

In and around Bidar, 65.74 per cent of donkeys had overall helminth infestations. Strongylus spp were more common among the helminth infection (51.76%) than Parascaris equorum spp (14.08%). Strongyloid spp prevalence was (7.04%), whereas Gastrodiscus spp prevalence was (7.74%). Oxyuris equi spp was (1.05%) and Anaplocephala spp was (0.70%) prevalent.

Acknowledgement

AcknowledgmentAuthors' hereby acknowledges the Veterinary College, Bidar for their support and facility.

References

1. Ajabe JS, Borikar ST, Siddiqui MFMF. Studies on prevalence of helminthosis in donkeys of certain districts of Marathwada region (Maharashtra State, India). J Entomol Zool Stud. 2020;8(2):1430-1432.
2. Ali TMO, Ibrahim KEE, Eltom EHA, Hamid ME. Animal disease diagnosed at the University of Khartoum Veterinary Teaching Hospital (1995-1998). Sud J Vet Sci Anim Husb. 2001;40:38-44.
3. Alshaibani I, AlDalali Z, Al-Shaibani I. Epidemiological study on gastrointestinal nematodes of donkeys in Dhamar Governorate, Yemen. Saudi J Med Pharm Sci. 2019;5(5):379-385.
4. Austin MS. Large strongyles in horses. Compend Contin Educ Pract Vet. 1994;16(5):650-657.
5. Ayele G, Feseha G, Bojia E, Joe A. Prevalence of gastro-intestinal parasites of donkeys in Dugda Bora District, Ethiopia. Livest Res Rural Dev. 2006;18(10):14-21.
6. Bhavanakumar CK, Satchidanandam V. Gastrointestinal parasitic worms in horses at Soorapet, near Madras. Centaur. 1987;3(3):103-104.
7. Chitra RS, Rajendran S, Prasanna D, Kirubakaran A. Influences of age on the prevalence of parasitic infections among donkeys in Erode District, Tamil Nadu, India. Vet World. 2011;4(6):258-259.
8. Esmaeel SA. Clinical and hematological study of the internal parasites in native donkeys in Mosul City. Iraqi J Vet Sci. 2010;24(2):129.
9. Hamed MI, El-Allawy TA, Hassnein E. Prevalence and anthelmintic resistance of strongyle infection of donkeys in El-Wadi El-Gadid, Egypt. J Adv Vet Res. 2019;9(4):144-150.
10. Herd RP. The changing world of worms: The rise of the cyathostomes and the decline of Strongylus vulgaris. Compend Contin Educ Pract Vet. 1990;12:732-736.
11. Herd RP, Coles GC. Slowing spread of anthelmintic resistant nematodes of horses in the United Kingdom. Vet Rec. 1995;136:481-485.
12. Ibrahim N, Berhanu T, Deressa B, Tolosa T. Survey of prevalence of helminth parasites of donkeys in and around Hawassa Town, Southern Ethiopia. Glob Vet. 2011;6:223-227.
13. Imam SMA, Hassan T, Seri HI, Zolaio BB. Field investigation of gastro-intestinal nematodes in horses and donkeys in South Darfur State, Sudan. In: 13th Scientific Congress, Faculty of Veterinary Medicine, Assiut University, Egypt; 2008. p. 723-729.
14. Kelly JD, Webster JH, Griffin DL, Whitlock HC, Martin ICA, Gunawan M. Resistance to benzimidazole anthelmintics in equine strongyles. Frequency, geographical distribution and relationship between occurrence, animal husbandry procedures and anthelmintic usage. Aust Vet J. 1981;57:163-171.
15. Kelly JC, Fogarty UMG. Outbreak of larval cyathostomiasis on a thoroughbred stud farm. Ir Vet J. 1993;46:133-136.
16. Khajuria JK, Yadav A, Raina AK. Incidence of helminth parasites in equines of Jammu Region. Centaur. 2004;21:58-61.
17. Khan A, Khan MS, Avais M, Mahmood AK, Ijaz M. Prevalence and chemotherapy of Parascaris equorum in equines in Pakistan. J Equine Vet Sci. 2010;30:155158.
18. Klei TR. Recent observations on the epidemiology, pathogenesis and immunology of equine helminth infections. In: Proceedings of the 6th International Equine Infectious Diseases Conference. R&W Publishing Ltd.; 1992. p. 129-136.
19. Lichtenfels JR. Helminths of domestic equids. Proc Helminthol Soc Wash. 1975;42:1-92.
20. Linde OE, Hogland J, Ljungstrom BL, Nilsson O, Uggla A. A field survey on the distribution of strongyle infections of horses in Sweden and factors affecting faecal egg counts. Equine Vet J. 1999;31:68-72.
21. Love S, Mair TS, Hillyer MH. Chronic diarrhea in adult horses: A review of 51 referred cases. Vet Rec. 1999;130:217-219.
22. Mahfooz A, Masood MZ, Yousaf A, Akhtar N, Zafar MA. Prevalence and antihelmintic efficacy of abamectin against gastrointestinal parasites in horses. Pak Vet J. 2008;28:76-78.
23. Mathewos M, Fesseha H, Yirgalem M. Study on strongyle infection of donkeys and horses in Hosaena District, Southern Ethiopia. Vet Med. 2021;12:67-73.
24. Molla B, Worku Y, Shewaye A, Mamo A. Prevalence of strongyle infection and associated risk factors in equine in Menz Keya Gerbil District, North-Eastern Ethiopia. J Vet Med Anim Health. 2015;7(4):117-121.
25. Nagar SR, Khatoon S, Alam HM, Purohit JK, Dixit AK. Epidemiological studies on gastrointestinal helminths of horses of Udaipur, Rajasthan. 2022.
26. Negash W, Erdachew Y, Dubie T. Prevalence of strongyle infection and associated risk factors in horses and donkeys in and around Mekelle City, Northern Part of Ethiopia. Vet Med Int. 2021;2021:1-7.
27. Mezgebu T, Tafess K, Tamiru F. Prevalence of gastrointestinal parasites of horses and donkeys in and around Gondar Town, Ethiopia. Open J Vet Med. 2013;3(6):267.
28. Ogbourne CP. Pathogenesis of cyathostome (Trichonema) infections of the horse: A review. Commonwealth Institute of Helminthology, Miscellaneous Publications. 1978. p. 25.
29. Parasani HR. Helminthic infections of donkey (Equus asinus) with special reference to strongylosis in North Gujarat [PG thesis]. Sardarkrushinagar: Sardarkrushinagar Dantiwada Agricultural University; 2008.
30. Parsani HR, Momin RR, Lateef A, Das H. Studies on gastro-intestinal helminths of Equus asinus in North Gujarat, India. Egypt J Biol. 2013;15:13-20.
31. Radostits OM, Gay CC, Hinchcliff KW, Constable PD. Veterinary medicine: A textbook of the diseases of International Journal of Advanced Biochemistry Research https://www.biochemjournal.com cattle, horses, sheep, pigs and goats. 10th ed. London; 2007. p. 1558-1562.
32. Sabir HM, Bandyopadhyay PK. Prevalence of gastrointestinal parasites in economically important Bonpala sheep in India. J Agri Vet Sci. 2016;9(1):87-93.
33. Seri HI, Hasan T, Salih MM, Abakar AD. A survey of gastrointestinal nematodes of donkey (Equus asinus) in Khartoum State, Sudan. J Anim Vet Adv. 2004;3(11):736-739.
34. Shadia A, Idriss BG, Hisham IS, Mohammed YM. Epidemiology of parasitic infection among working donkeys in Qubaish Locality, Western Kordufan State, Sudan. 2018.
35. Shah-Fischer M, Say RR. Manual of Tropical Veterinary Parasitology. Wallingford: CAB International; 1989. p. 410.
36. Snedecor GW, Cochran WG. Statistical Methods. 8th ed. Ames (IA): Iowa State University Press; 1994. p. 591.
37. Soulsby EJL. Biology of Parasites. New York and London: Academic Press; 1966. p. 185.
38. Soulsby EJL. Helminths, Arthropods and Protozoa of Domesticated Animals. 7th ed. London: Baillière Tindall; 1982. p. 172-179.
39. Takele B, Nibret E. Prevalence of gastrointestinal helminths of donkeys and mules in and around Bahir Dar, Ethiopia. Ethiop Vet J. 2013;17(1):13-30.
40. Taye A. Epidemiological study on equine gastrointestinal helminth parasites in Mekelle, North Ethiopia. Open J Vet Med. 2017;7(10):121.
41. Tudor P. Study regarding the prevalence of endoparasitary infestation of donkeys from a shelter. Sci Works Univ Agron Sci Vet Med Bucharest Ser C Vet Med. 2010;56(3-4):453-457.
42. Uhlinger CA. Uses of faecal egg count in equine practice. Compend Contin Educ Pract Vet. 1993;15:742-748.
43. Waqas M, Khan MS, Durrani AZ, Khan MA, Avais M, Khan SA, Rehman SU, Hussain A, Nasir A, Hussain A, Santos FC. Prevalence of gastrointestinal parasites, chemotherapy and haematology of strongylosis in donkeys of District Lahore, Pakistan. Int J Curr Microbiol App Sci. 2014;3(7):198-207.
44. Wheat JD. Causes of colic and types requiring surgical intervention. J S Afr Vet Assoc. 1975;46:95-98.
45. White NA, Moore JN, Douglas M. SEM study of Strongylus vulgaris larvae-induced arteritis in the pony. Equine Vet J. 1993;15:349-353.