ReseaRch aRticle Journal of Agricultural and Marine Sciences 2021, 26(2): 51–55 DOI: 10.24200/jams.vol26iss2pp51-55 Reveived 24 July 2020 Accepted 17 Dec 2020 Seroprevalence of Trypanosoma evansi infections among dromedary camels (Camelus dromedaries) in North Ash-Sharqiya Governorate, Sultanate of Oman A. H. Al-Kharusi1, Elshafie I. Elshafie2,5*, K. E. M. Ali4, R. Al-Sinadi1, N. Baniuraba3, F. Al-Saifi1 and Y. Al-Mawali1 Elshafie I. Elshafie2,5*( ) eielshafie@squ.edu.om, 1Ministry of Ag- riculture, Fisheries Wealth and Water Resources, Sultanate of Oman, 2Department of Animal and Veterinary Sciences, College of Agri- cultural and Marine Sciences, Sultan Qaboos University, Muscat PC. 123, Sultanate of Oman, 3Nizwa University, Sultanate of Oman, 4Veterinary Pharmacy, Abu Yahya AlAbrawi Trad, Sultanate of Oman, 5Central Veterinary Research Laboratory, Al Amarat, P.O. Box 8076, Khartoum, Sudan. Introduction Parasite remains a major constraint for optimum animal production throughout the world. Try-panosoma evansi, which belongs to the genus Try- panosoma, subgenus Trypanozoon is a blood parasite that infects domestic livestock and wild animals; and caused disease called surra (Bargash et al., 2014). The parasite has a wide geographical distribution in many االنتشار املصلي لعدوى امِلثـَْقِبيَّة اإليفانسيَّة بني اإلبل وحيدة السنام يف حمافظة مشال الشرقية، سلطنة عمان أمل محد اخلروصية، الشفيع ابراهيم الشفيع، كمال على، راشد السنيدي، اندية بين عرابة، فاطمة السيفية، حيىي املعويل Abstract. Trypanosoma evansi is a well-known hemoprotozoa that infects diverse domestic and wild animals world- wide and causes a devastating disease called surra. This research aimed to investigate seroprevalence of Trypanosoma evansi and its associated risk factors in dromedary camels in North Ash-Sharqiya Governorate, Sultanate of Oman. A total of 4364 serum samples was collected from three Willayat in North Ash-Sharqiya Governorate of Sultanate of Oman; Ibra (926, 21.2%), Qabil (1119, 25.6%) and Bidiyah (2319, 53.1%). Samples were examined for the presence of antibodies against T. evansi using card agglutination test for T. evansi (CATT/T. evansi). Binary logistic regression was used to study the association of T. evansi seroprevalence and risk factors such as location, gender, purpose and age of camels. The overall seroprevalence of T. evansi detected by CATT/T. evansi test was 38% (1659/4364, CI: 36.6-39.5%). There was a significant difference (p=0.001) between location and T. evansi seroprevalence, whereas the highest sero- prevalence was found in Ibra (49.9%, CI: 46.7-53.1%) followed by Bidiyah (35%, CI: 33-36.9%) and Qabil (34.5%, CI: 31.8-37%). Camels from Ibra were almost two times more likely to have circulating antibodies of T. evansi than camels from Bidiyah (OR=1.89, CI: 1.591-2.168). The results of this study showed a significant difference between seropreva- lence and sex (p=0.023), whereas the age of camels did not show any significant difference (p>0.05). To our knowledge, this is the initial research that indicated that T. evansi antibodies were circulating among camels in Oman, and further research needs to be tackled to study the molecular characterization of T. evansi and its prevalence in other animal species. Furthermore, cross-sectional studies of T. evansi from different regions in Oman warrant further investigation. Keywords: Camels, T. evansi, CATT, Seroprevalence, Oman امللخص:املِثـَْقِبيَّــة اإليفانســيَّة هــو نــوع معــروف مــن األوليــات وحيــدة اخلليــة يصيــب احليــواانت األليفــة والربيــة املتنوعــة يف مجيــع أحنــاء العــامل ويســبب وحيــدة اإلبــل يف هبــا املرتبطــة اخلطــر وعوامــل اإليفانســيَّة للمثقبيــات املصلــي االنتشــار دراســة إىل البحــث هــذا يهــدف الزنبــور. يســمى مرًضــا الســنام مبحافظــة مشــال الشــرقية، ســلطنة عمــان. مت مجــع 4364 عينــة مــن مصــل الــدم مــن ثــاث والايت يف حمافظــة مشــال الشــرقية ســلطنة عمــان. إبــراء )926، ٪21.2(، القابــل )1119، ٪25.6(، بديــة )2319، ٪53.1( مت فحــص وجــود األجســام املضــادة ضــد املِثـَْقِبيَّــة اإليفانســيَّة ابســتخدام اختبــار تــراص البطاقــة للِمثـَْقِبيَّــة اإليفانســيَّة )CATT(. مت اســتخدام االحنــدار اللوجســي الثنائــي لدراســة االرتبــاط بــن االنتشــار املصلــي للِمثـَْقِبيَّــة اإليفانســيَّة وعوامــل اخلطــر مثــل املوقــع واجلنــس والغــرض والعمــر لإلبــل. كان االنتشــار املصلــي الكلــي للِمثـَْقِبيَّــة اإليفانســيَّة املكتشــف بواســطة اختبــار تــراص البطاقــة للِمثـَْقِبيَّــة اإليفانســيَّة )CATT( 38% (1659/4364, CI: 36.6-39.5%(. كان هنــاك فــرق إحصائــي )p = 0.001( بــن املوقــع واالنتشــار املصلــي )CI: 33-36.9٪ ،35٪( تليهــا بديــة )CI: 46.7-53.1٪ ،49.9٪( للِمثـَْقِبيَّــة اإليفانســيَّة، حيــث مت العثــور علــى أعلــى معــدل انتشــار مصلــي يف إبــراء والقابــل )CI: 31.8-37٪ ،34.5٪(. كانــت اإلبــل مــن إبــراء أكثــر عرضــة مبرتــن لوجــود األجســام املضــادة املنتشــرة مــن املِثـَْقِبيَّــة اإليفانســيَّة مقارنــة جبمــال بديــة )OR = 1.89، CI: 1.591-2.168(. أظهــرت النتائــج يف هــذه الدراســة فرقــاً احصائيــا بــن االنتشــار املصلــي واجلنــس )p = 0.023(، بينمــا مل يظهــر عمــر اإلبــل أي فــرق إحصائــي )p> 0.05(. علــى حــد علمنــا، هــذا هــو البحــث األويل الــذي أشــار إىل أن األجســام املضــادة للِمثـَْقِبيَّــة اإليفانســيَّة توجــد بــن اإلبــل يف عمــان، وهنــاك احلاجــة إىل املزيــد مــن البحــث لدراســة خصائــص املِثـَْقِبيَّــة اإليفانســيَّة وانتشــارها يف أنــواع احليــواانت األخــرى، عــاوة علــى ذلــك، يتطلــب مزيــدا مــن البحــث والدراســات للِمثـَْقِبيَّــة اإليفانســيَّة يف مناطــق خمتلفــة مــن عمــان. الكلمات املفتاحية: اإلبل، املِثـَْقِبيَّة اإليفانسيَّة، اختبار تراص البطاقة، االنتشار املصلي، سلطنة عمان 52 SQU Journal of Agricultural and Marine Sciences, 2021, Volume 26, Issue 2 Seroprevalence of Trypanosoma evansi infections among dromedary camels (Camelus dromedaries) in North Ash-Sharqiya Governorate, Sultanate of Oman tropical and subtropical countries due to the adaptation of mechanical transmission by different types of hema- tophagous flies, such as Tabanus, Stomoxys, Lyperosia and Haematobia species (Desquenes et al., 2013). The disease was reported in camels from different regions of the Kingdom of Saudi Arabia (AlTaqi et al., 2011; Amoudi et al., 2011; El-Wathiq et al., 2016), in Kuwait (Al-Taqi, 1989) and United Arab Emirates (Chaudhary and Iqbal, 2000). Camels along with horses and dogs are considered the principal hosts to the parasite and this may show symptoms, such as fever, anorexia, and edema (Abdel-Rady, 2008). The disease has a significant impact on animal’s health, production and efficiency and can result in death of animals if no timely treatment is offered. Some biochemical changes have been docu- mented in camels related to Trypanosoma infection in dromedary camels. (Sazmand et al., 2011). The disease causes infertility in the dromedary camels by changing plasma steroids concentration and semen characteris- tics (Al-Qarawi et al., 2004). The disease can be diagnosed by parasitological methods, such as microhematocrit centrifugation technique and microscopic examination of thin blood smears (Babeker and Hassab, 2014; Bennoune et al., 2013; Bogale et al., 2012; Hagos, 2009) and serological methods such as ELISA and card agglutination test for T. evansi (CATT/T. evansi) (Babeker et al., 2014; Zayed et al., 2010; Hagos, 2009). The introduction of molecular diagnosis enhanced the accuracy of T. evansi detection due to the dynamics of parasitaemia (Baraghash et al., 2014; Tehseen et al., 2015). The Card agglutination test was more common to be used for screening a large num- bers of animals under field conditions (Uilenberg, 1998). On the other hand, the CATT test can render false posi- tive results due to the circulation of antibodies. The estimated number of camels in the Sultanate of Oman is 242833 heads (Agriculture Censuses, 2013). The highest density of the camel population in Oman is located in the Dhofar Governorates, as most people use camels for milk and meat production. The second highest density of camels is in Ash-Sharqiya North Gov- ernorate with a total of 21577 camels and mostly are raced camels. Since the first detection of the parasite in 1984 (Srivastava,1984) and the continuing reports of the disease from the Ministry of Agriculture, Fisheries Wealth and Water Resources, Trypanosoma evansi was wandering in the country and negligible cohort study has been done to identify the frequency and risk factors of the disease in livestock. Due to lack of work on se- roprevalence of Trypanosoma among camel population in Oman, the present study was planned to determine the seroprevalence and risk factors of T. evansi in Omani camels in North Ash-Sharqiya Governorate using sero- logical techniques. Materials and Methods Study Area and Sampling Collection This study was conducted in three Willayat in North Ash-Sharqiyah (Ibra, Qabil and Bidiyah). A total of 4364 camel blood samples were collected between July 2014 and November 2015 from Ibra (283 farms), Qabil (260 farms) and Bidiyah (394 farms). Ten millilitres of blood was collected from jugular vein of each camel in a plain tube and transported in a cool box to the lab for screen- ing T. evansi using card agglutinin test (CATT/ T. evan- si). Information on location, sex, age and camel purpose were collected during blood sample collection. In this study, camels were divided into three groups according to age (<5, 5-10 and >10 years old). Diagnosis of Serum by Serological Examination Card agglutination test for T. evansi kit (CATT/ T. evan- si) was used to detected anti-trypanosome antibodies in serum of infected animal by direct agglutination method according to the manufacturer’s instructions (Institute of Tropical Medicine, Antwerp, Belgium). The antigen contains fixed, freeze-dried and stained bloodstream from trypanosomes type RoTat 1.2, a predominant variable antigen type of Trypanosome evansi expressed during the early course of infection (Bajyana-Songa and Hamers, 1988). About 2.5 ml of CATT/T. evansi buffer was added to a vial of freeze dried CATT/ T. evansi anti- gen and shake the vial immediately for a few seconds to obtain homogeneous suspension. Then 0.5 ml of CAT- T/T. evansi buffer was added to a positive and negative control. On the test area of the card, one drop (45 μl) of the homogenized CATT/T. evansi antigen was added in each test area and 25 μl of the serum diluted at 1:4 with PBS pH 7.2 according to the manufacturer’s instructions was then added. The reaction was mixed using a stirring rod and the test card was rotated for 5 minutes for sub- sequent reading of the results. Statistical Analysis The statistical analysis was conducted using SPSS version 20 (IBM, SPSS) at a = 0.05 significance level. Seropreva- lence and associated risk factors were conducted at 95% confidence level. Univariate analysis for individual risk factors to identify association between the seroprevalence and the potential risk factors were tested using Pearson Chi-square or Fisher Exact test. A binary logistic regres- sion was used to examine the significance that revealed by the univariate analysis and to determine odds ratio. Results and Discussion Out of 4364 blood samples examined using CATT/T. evansi, 1659 (38%, CI: 36.6-39.5%) were found to be se- ropositive for T. evansi (Table 1). The frequency of in- fection was high in Ibra 49.9% (462/926, CI: 46.7-53.1%) 53Research Article Al-Kharusi, Elshafie, Ali, Sinadi, Baniuraba, Al-Saifi, Al-Mawali followed by Bidiyah 35% (811/2319, CI: 33-36.9%) and Qabil 34.5% (386/1119, CI: 31.8-37%). There was sig- nificant difference between disease seroprevalence and location (x2 =70.43, p=0.001) (Table 2). Based on logistic regression, camels that sampled from Ibra region were 1.89 times more likely to have circulating antibodies against T. evansi than camels sampled from Bidiyah re- gion (OR=1.89, 95%CI: 1.591-2.168). The results showed a significant difference between seroprevalence and sex (p=0.023). The female had high- est infection rate 38.7% (1435/3706) than male 34% (224/658). The female seropositive camels were about 1.2 times more likely than male camels. Moreover, there was no significant association between diseases seropreva- lence and the purpose (p=0.053). Despite the non-sig- nificance difference, camels used for production 38.6% (1488/3858) had a higher rate of being seropositive than racing camels 34.1% (168/493). On the other hand, there was no statistical significant difference between diseases seroprevalence and the age (p>0.05) (Table 1). To our knowledge, this is the initial study that pro- vides information about the seroprevalence of T. evansi infections in three regions in Oman using serological test (CATT/T. evansi). The results showed significant difference between diseases seroprevalence and camel sex and sample location. On the other hand, there is no significant difference between the diseases seropreva- lence with camel age and purpose of camels. The overall seroprevalence of T. evansi infections was found to be 38%. Our finding is relatively lower than the results re- ported in Saudi Arabia with a seroprevalence of 39.4% by card agglutination test with considerable differences between eastern and central regions (Al-Afaleq et al., 2015). The seroprevalence of this study was higher than other studies done in Ethiopia by Bogale et al. (2012), Birhanu et al. (2015), Hagos et al. (2009), and in Somalia by Mohamoud (2017), who reported a prevalence of T. evansi in camels 18.22%, 13.76%, 24.88%, 15.9% respec- tively. However, our results of T. evansi seroprevalence was relatively lower than that reported in Sudan and Egypt as estimated to be 52.2% and 43.5% respectively (Babeker and Hassab Elrasoul, 2014; Abdel-Rady 2008). Since CATT/T. evansi cannot differentiate between cur- rent and past infections, the higher seroprevalence of T. evansi revealed by this research may be attributed either to genetic variation of camels breeds existed in same farms or weak farm management programs adopted by camel’s owner as the causative agent was reported in the country without comprehensive study to reveal its distribution. Trypanosoma evansi seroprevalence was higher in Ibra (49.9%) than Qabil and Bidiyah, this might be due to environmental factors as Ibra is a mountainous area and had wetland that may enhance the propagation of the parasite vectors, whereas Qabil and Bidiyah are sand areas with significantly less vector density. In the present study, a significant difference was reported in camel sex, the highest seroprevalence of Trypanosoma evansi was observed in female 38.7% fol- lowed by male 34%. This result in accordance with that found by Babeker and Hassab Elrasoul (2014) and Mo- hamoud (2017). However, our results are not in line with study done by Bogale et al. (2012) which showed that a higher infection rates in males (20.25%) compared to female (17.72%) with approximately similar prevalence rates among both sexes. Study conducted by Bhutto et Table 1. : Association of location, age and sex with seroprevalence of Trypanosoma evansi using card agglutination test (CATT/ T. evansi). Risk factors No. CATT/ T. evansi Chi-square P-value Positive (%) Negative (%) Location Ibra Qabil Bidiyah 926 1119 2319 462 (49.9) 386 (34.5) 811 (35) 464 (50.1) 733 (65.5) 1508 (65) 70.43 0.001* Age (years) <5 5-10 >10 1555 2152 649 608 (39.1) 811 (37.7) 237 (36.5) 947 (60.9) 1341 (62.3) 412 (63.5) 1.49 0.47 Sex Male Female 658 3706 224 (34) 1435 (38.7) 434 (66) 2271 (61.3) 5.19 0.023* Purpose Race Production 493 3858 168 (34.1) 1488 (38.6) 325 (65.9) 2370 (61.4) 3.74 0.053 *Significant association (p<0.05) 54 SQU Journal of Agricultural and Marine Sciences, 2021, Volume 26, Issue 2 Seroprevalence of Trypanosoma evansi infections among dromedary camels (Camelus dromedaries) in North Ash-Sharqiya Governorate, Sultanate of Oman al. (2010) indicated that females were more suscepti- ble to the infection with Trypanosoma species as host immunity decreased during pregnancy and lactation period. Despite the non-significant result of the age, we observed an increase in T. evansi seroprevalence in younger camels with age less than five years (39.1%) as compared to camels with age between five and ten years (37.7%). This study disagreed with previous studies, which found that adult camels were more susceptible to have the diseases than younger camels (Mohamoud, 2017; Atarhouch et al., 2003). Detection of antibodies against T. evansi in the host does not necessarily indi- cate a current infection since antibodies can persist for 2.3–22.6 months after trypanocidal treatment (Monzón et al., 2003). Therefore, younger camels of age less than five years have a higher chance of encountered antibod- ies against T. evansi in their blood compare to the elder one. In addition, younger camels are used to transport to different places in country for racing purposes and exposing these animals to infections. Camels used in production showed higher percentage of T. evansi sero- prevalence than camels using for race 34.1% and 38.6%, respectively. This might be because camel’s keepers care more about racing camels and keep them in open area for grazing, provide them with sufficient complementa- ry diet and treat them against parasites whereas, camels that used for production get less care from camel owner. This could also be due to disparity in management, vet- erinary care, and parasite control offered to race camels compared to production animals. Conclusion To our knowledge, this research was the initial report on T. evansi infections in Oman by using card agglutina- tion test (CATT/T. evansi). The results showed that the T. evansi was circulating in camels within the investigat- ed areas and certain potential risk factors may be asso- ciated with the spreading of the disease in the country. The disease might cause great economic losses due to impairment of camel’s health, which leads to decrease in production and performance. This study was done in North Ash-Sharqiya Governorate of Sultanate of Oman, so further studies are highly needed to investigate the prevalence of the diseases in different parts of Oman in different livestock using different diagnostic techniques. Acknowledgement We would like to thank the Agriculture Development Fund for supporting the project and all staff of General Directorate of Agriculture and Animal wealth of North and South Al-Sharqiya Governorate for their collabora- tion in samples collection. Conflict of interest The authors declare that they have no conflict of interest. References Abdel-Rady A. (2008). Epidemiological studies (parasito- logical, serological and molecular techniques) of Try- panosoma evansi infection in camels (Camelus drom- edaries) in Egypt. Veterinary World 1(11): 325-328. Agriculture Censes. (2013). 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