Survey on prevalence of blood parasites on beef cattle in cho moi district – An Giang province

The study was conducted in Cho Moi district, An Giang province and aimed to determine the prevalence of blood parasites in beef cattle. Design of the study was a cross-sectional with a total of 339 beef blood samples collected to test for the presence of blood parasites by using of Giemsa-stained thin blood smear method. The results showed that the prevalence of blood parasites in beef cattle was 13.57%. Two species were identified, Babesia bigemina and Anaplasma margina. The infected rate of Babesia bigemina and Anaplasma margina were 6.20% and 4.72%, respectively and multiple infection of both Babesia bigemina and Anaplasma margina was 2.65%. Boophiplus microplus and Rhipicephalus sanguineus tick were 80.00% and 19.92%, respectively, among 231 samples of tick. Relative risk ratio was 4.76. This result showed that the exposure factor of blood parasites highly related with vectors of Boophiplus microplus and Rhipicephalus sanguineus with correction is 4,76.

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An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 19 SURVEY ON PREVALENCE OF BLOOD PARASITES ON BEEF CATTLE IN CHO MOI DISTRICT – AN GIANG PROVINCE Nguyen Phi Bang1, Nguyen The Thao1, Le Thi Thuy Hang1 1An Giang University Information: Received: 02/01/2016 Accepted: 01/03/2016 Published: 06/2017 Keywords: Blood parasite, Babesia bigemina, Anaplasma margina, Boophiplus microplus, Rhipicephalus sanguineus ABSTRACT The study was conducted in Cho Moi district, An Giang province and aimed to determine the prevalence of blood parasites in beef cattle. Design of the study was a cross-sectional with a total of 339 beef blood samples collected to test for the presence of blood parasites by using of Giemsa-stained thin blood smear method. The results showed that the prevalence of blood parasites in beef cattle was 13.57%. Two species were identified, Babesia bigemina and Anaplasma margina. The infected rate of Babesia bigemina and Anaplasma margina were 6.20% and 4.72%, respectively and multiple infection of both Babesia bigemina and Anaplasma margina was 2.65%. Boophiplus microplus and Rhipicephalus sanguineus tick were 80.00% and 19.92%, respectively, among 231 samples of tick. Relative risk ratio was 4.76. This result showed that the exposure factor of blood parasites highly related with vectors of Boophiplus microplus and Rhipicephalus sanguineus with correction is 4,76. 1. INTRODUCTION Cho Moi is an alluvial island consolidated by Tien river and Hau river. Here the fertile land is suitable for rice and dryland crops production, these are also the main sources of agricultural products in beef cattle breeding. In Cho Moi district of An Giang province, beef cattle breeding is very developed, and the district also has incentive loans for livestock development in order to encourage household or farm animal husbandry (Le Thang, 2014). This policy helps develop the beef cattle herd in Cho Moi district and contribute to raise income for farmers. However, the majority of herders in Cho Moi district raise beef cattle, and they frequently exchange or trade beef cattle with those in other areas. These trading and exchange from one area to another are key factors to the spread of diseases. Though they do not cause a massive pandemic as infectious diseases, diseases caused by parasites have complicated development, spread out silently and smoulderingly, and finally create a significant loss to the livestock industry. Diseases with the highest impact are the blood parasite diseases, i.e the Trypanosomiasis, Babesiosis, Anaplasmosis and Theileriosis (Nguyễn Hữu Hưng, et al., 2015). They fall under the list of dangerous diseases to animals (MARD, 2005). Findings of the research can help veteranians evaluate the circulating levels of blood parasite diseases on beef cattle in Cho Moi district, An Giang province. 2. MATERIALS AND METHODOLOGY 2.1 Materials Research object is the beef cattle in the age <1 year, 1-2 years, > 2 years at household farms in Cho Moi district of An Giang province. An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 20 Materials, instruments and chemicals used in the experiment: microscopy, lame, lamelle, specimen containers, mice, Giemsa dye, Ethanol, Methanol, Glycerol, Xylene, antifreeze, microscopic examination oil. 2.2 Research methodology 2.2.1 Survey methodology By the method of cross-sectional study and evidence-control study (understanding the relation between one (or more) risk factors and blood parasitic diseases). 2.2.2 Sampling and sample storing methodology Blood samples of beef cattle were taken randomly at investigated areas. The diagnostic methods are based on the method of (Pham Sy Lang et al, 2006). Get the venous blood in the ear fringe of beef cattle, Giemsa-stained thinly spread method of dry blood to find the presence of blood parasites. Infusion over white mice to find the presence of Trypanosoma. Ticks and broking insect vectors were identified and classified based on the identification method of authors: Trinh Van Thinh (1982), Phan Trong Cung (1977). The intensity of blood parasitic infections on beef cattle is rated at three different levels, low intensity (mild infection "+"): There are 1-2 parasites/environment; The medium intensity (moderate infection "++"): There are 3-4 parasites/environment; high intensity (severe infection "+++"): over 4 parasites/environment (Ho Thi Thuan et al, 2000). 2.2.3 Data processing methodology Calculate prevalence rate in Excel and compare prevalence rate of blood parasites by Chi-Square test of statistical software Minitab version 13.1. 2.2.4 Relation ratio between risk and disease Using RR ratio (Relative ratio) for evaluating association level between blood parasitic disease and rate of ticks infection Michael Thrusfield (2007). 3. RESULTS AND DISCUSSION 3.1 The prevalence of blood parasitic infection on beef cattle in Cho Moi district – An Giang province Table 1. Rate of blood parasitic infection at communes of Cho Moi district Investigate d area No. of animals (cattle) No. of infecte d animal s (cattle) Infection rate (%) Babesia Bigemina Anaplasma marginal Graft infection No. of infected animals (cattle) Infection rate (%) No. of infected animals (cattle) Infection rate (%) No. of infected animals (cattle) Infection rate (%) My An 90 13 14.44 6 6.67 5 5.56 2 2.22 Nhon My 80 10 12.50 4 5.00 3 3.75 3 3.75 Kien An 99 16 16.16 8 8.08 6 6.06 2 2.02 My Hoi Dong 70 7 10.00 3 4.29 2 2.86 2 2.86 Total 339 46 13.57 21 6.20 16 4.72 9 2.65 An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 21 By Giemsa-stained method and infusion through mice we found no presence of Trypanosoma evansi and Theileria on 339 tested samples, this result might be due to cattle here have fewer infection rates with very low intensity or cattle in areas were not infected or because the survey area did not have this disease. Results in Table 1 show beef cattle in four surveyed communes were infected with blood parasites and disease prevalence is high or low depending on the locality. At Kien An commune, we noted the proportion of blood parasite infection is highest 16.16%, followed by My An commune with a ratio of 14.44%, at Nhon My commune 12.50% and My Hoi Dong commune had the lowest prevalence rate, 10%. Thus, the study showed that overall prevalence of blood parasites on the herd of beef cattle at Cho Moi district with general infection rate was 13.57% on the total number of beef cattle being investigated. Statistical analysis indicated difference in the prevalence of blood parasites in four investigated communes at Cho Moi District has no statistical significance. This can be explained by the communes located in the same ecological zone, climate, identical grazing conditions, similar care. Additionally, through interviewed farmers, most of them at Cho Moi district have similar points with farming methods, food, drinking water, sanitation, cleaning and breeding waste treatment which makes the infection rate different but not statistically significant. Findings of the level of blood parasitic infection on beef cattle has low overall infection rate than the domestic research of Nguyen Huu Hung et al (2014) in two districts Tinh Bien and Tri Ton, but much lower than the research of Imelda Kartini Tefi et al (2014) when examining the prevalence of blood parasites on beef cattle imported into Australia. The reason for this difference is because the majority of beef cattle at Cho Moi District are household and well-cared, so the spread of this disease is greatly limited. At other survey areas, most of cattle are grazing, this is the main reason for the greater possibility of spreading blood parasite disease that leads to higher infection rate. 3.2 The prevalence of blood parasitic infection on beef cattle by age in Cho Moi district – An Giang province Table 2. The prevalence of blood parasites on beef cattle by age Age No. of animals (cattle) No. of infectedanimals (cattle) Infection rate (%) Babesia Bigemina Anaplasma marginal Graft infection No. of infectedani mals (cattle) Infectio n rate (%) No. of infectedanim als (cattle) Infectio n rate (%) No. of infectedanim als (cattle) Infectio n rate (%) < 1 year 213 9 4.23a 7 3.29 1 0.47 1 0.47 1-2 years 87 18 20.69b 5 5.75 8 9.19 5 5.75 > 2 years 39 19 48.72c 9 23.08 7 17.95 3 7.69 Total 339 46 13.57 21 6.20 16 4.72 9 2.65 Note: if a, b, c in the same column are different, that is statistically difference (P-value =0,001 The results in table 2 show that cattle in the survey got blood parasites at all 3 ages (<1 year; 1-2 years; >2 years) with different rates of infection. The results of statiscal analysis also showed the significant difference about rate of infection by age. Research recorded a prevalence of blood parasites in cattle at the age of under 1 year is 4.23% An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 22 (infected with Anaplasma margina is 0.47%, infected with Babesia bigemina is 3.29%, graft infection is 0.47%). At the age of 1-2 years, infection rate is 20.69% (in which, 5.75% infected with Babesia bigemina, 9.19 % infected with Anaplasma Margina, with 05 cases of Babesia bigemina and Anaplasma Margina graft infection, in a percentage of 5.75%). For cattle with the age >2 years, infection rate is 48.72% (in which, 23.08% infected with Babesia bigemina, 17.95% infected with Anaplasma margina, with 03 cases of Babesia bigemina and Anaplasma Margina graft infection, in a percentage of 7.69%). The data table also shows that surveyed cattles that were infected with blood parasites were mainly in the ages of 1- 2 years and > 2 years, prevalence of these parasites has tended to increase with age, this result is due to the same ecological conditions, older cows are exposed to opportunities in contacting with ticks and broking insects in higher pathoge, so the possibility of infection also increases with age. The research results are also consistent with the comments of Trinh Van Thinh (1978), Pham Sy Lang, Phan Đich Lan (2001). 3.3 Composition and intensity of blood parasitic infection on cattle in Cho Moi district – An Giang province Table 3. Composition and intensity of blood parasitic infection on cattle in Cho Moi district – An Giang province Classificatio n of parasites Number of infected animals n = 339 (cattle) Infectio n ratio (%) Intensity of infection (+) (++) (+++) No. of infected animals (cattle) Infection ratio (%) No. of infected animals (cattle) Infection ratio (%) No. of infected animals (cattle) Infectio n ratio (%) Babesia bigemina 21 6.19a 11 52.38 08 38.00 2 9.52 Anaplasma margina 16 4.72ab 12 75.00 04 25.00 0 0.00 Graft infection 09 2.65b 06 66.67 03 33.33 0 0.00 Total 46 13.57 29 63.04 16 34.78 2 0.00 Note: if a,b in the same column are different, that is statistically difference (P-value = 0,02) The results in Table 3 indicate that the prevalence of blood parasites with light intensity "+" is the highest (Babesia bigemina 52.38%; Anaplasma margina 75.00%). Next is medium level "++'' (Babesia bigemina 38.00%; Anaplasma margina 25.00%). Finally, the infection in severe level "+++" has lowest proportion (Babesia bigemina 9.52%; Anaplasma Margina 0.00%). On-fram observed symtoms and result from the subclinical examination showed the presence of blood parasites. However, cattles in light and medium intesity group had no clinical signs during sampling process. the result is consistent with finding Imelda Kartini Tefi et al, (2014).Authors such as Pham Van Khue, Phan Luc (1996) also confirmed no typical symptoms for blood parasites in light and medium infection intensity, often with chronic diseases and carrying parasite, they have the role of stockpiling and transfering pathogens in the nature . The cattle which carry parasites are a potential source of spreading germs, if all the factors, the disease will be found, such as the ecological conditions in place that facilitate the ticks, flies, swarms, bushy plants, damp, cold climate, cattle work hard, hungry, poor care ... The results are consistent with the study of Vo Thi Kim Mai (2008) An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 23 3.4 The prevalence of broking insects Table 4. Infection rate with broking insects The results in Table 4 show that the majority is Boophiplus microplus in surveyed areas, with a percentage over 80% of the parasitic form that we collected with appearance frequency of 185/231, while the frequency of Rhipicephalus sanguineus ticks is much lower (46/231) accounting for 19.92 %, which shows the adaptation and development of them in the survey area. It will be a big threat if the population of ticks increases , the possibility of spreading of blood parasites will also increase. Above results also show a huge role of ticks in the spread of blood parasitic disease in cattle. The survey showed infection with ticks accounted for 69.57% of cattle blood parasites. Among that, Boophilus microplus tick infection rate with 80.08% is the highest, followed by Rhipicephalus sanguineus ticks with high proportion of 19.92%, the study showed no presence of swarms Tabanus on the survey area. This result is consistent with the study of Phan Trong Cung (1985), Truong Le Van (2012), which also suggested that Boophiplus microplus tick is one of 44 species of ticks in Vietnam causing common parasitic disease, and has a very wide distribution with essential role in transfering Bovine Babesiosis disease. In addition to the role as spreading vector with blood parasitic disease, ticks have a very big impact on the health of cattle. During the blood sucking, they not only take away nutrients but also constitute skin wounds which are the entry for other pathogenic microorganisms, and that's a huge impact on the health and vitality of cattle. 3.5 Determine the risk relative between risk factor and blood parasitic disease Table 5. Risk relative between risk factors and blood parasitic disease Result Tick infection (E+) No tick infection (E-) Total RR No. of infected cattle 32 14 46 4.76 No. of non-infected cattle 78 215 293 Total 110 229 339 According to table 5, the number of cow catch infected tick with blood parasitic disease have ratio 32/46 holding 69.57%, the number of cow catch disease but non infected tick have ration 16/46 holding 30.43%, the number of cow non infected tick and not catch blood parasitic disease have ratio 215/293 holding 73.37% and the number of cow infected tick and not catch blood parasitic disease have ratio 78/293 holding 26.62%. The results represent the number of ticks infected cattle are at risk for blood parasitic diseases much higher than non -infected cattle ticks .The data in table 5 and PR index (relative risk) is 4.76 and we have concluded that: The exposure factor of vector Boophiplus microplussp and Rhipicephalus sp relate to blood parasitic disease. Above result is in conformity with Vo Thi Kim Mai’s comment (2008) cliamiing that ticks are the main factor to Name of broking insect Appearance frequency Rate (%) Boophilus microplus 185/231 80.08 Rhipicephalus sanguineus 46/231 19.92 Total 231 100 An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 24 transmit blood parasitic disease on cattle especially Babesia bigemina. 4. CONCLUSION Through tests on 339 samples of beef in Cho Moi District of An Giang Province, we recorded 46 samples of blood parasite infections with infected ratio as 13.57%. All four surveyed commnues were infected blood parasitic Babesia bigemina and Anaplasma margina. The ratio infected blood parasitic that have trend to increase with age. There are two types of infected parasite that are detected in the study as Boophiplus microplus và Rhipicephalus sanguineus, in there Boophiplus microplus hold majority (80.08%) and blood parasites in beef with intensity (+) percentage and (++) majority infected samples. Exposure factor of Boophiplus microplussp aned Rhipicephalus sp vector relate blood parasitic disease and correlation factor (relative risk) of the risk factor to affect disease as PR = 4.76. Hình 1. Babesia sp and Anaplasma sp with objective lens 100 Hình 2. Female Boophiplus sp and male Boophilus sp on beef cattle REFERENCES Imelda Kartini Tefi, Fadjar Satrija, & Umi Cahyaningsih. (2014). Study the Existence of Blood Parasites (Anaplasma, Babesia, Theileria) and Physiological Profiles of Australian Imported Feeder Cattle. Acta Parasitologica Globalis 6 (1): Page 55-59, ISSN 2079-2018. An Giang University Journal of Science – 2017, Vol. 5 (2), 19 – 25 25 Michael Thrusfield. (2007). Veterinary Epidemiology. 3rd ed. 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