Associated risk factors and management practices influence on Babesia prevalence in cattle of District Toba Tek Singh, Punjab, Pakistan

Current Science Perspectives 1(1) (215) 26-32 Associated risk factors and management practices influence on Babesia prevalence in cattle of District Toba Tek Singh, Punjab, Pakistan Munawar Iqbal 1, Tauseef-ur-Rehman 2, *, M. Nisar Khan 2, Zafar Iqbal 2, M. Sohail Sajid 2, Rao Zahid Abbas 2, M. Tariq Javed 3 and Muhammad N. Abbas 4 1 National Centre of Excellence in Physical Chemistry, University of Peshawar, Peshawar-2512, Pakistan 2 Department of Parasitology, University of Agriculture, Faisalabad-384, Pakistan 3 Department of Pathology, University of Agriculture, Faisalabad-384, Pakistan 4 Department of Zoology, University of Gujrat, Pakistan Corresponding author E-mail: drtauseef441@gmail.com A R T I C L E I N F O A B S T R A C T Article type Research article Article history Received 2 July 213 Accepted 1 December 214 Published 6 January 215 January 215 issue Keywords Parasitic diseases Infectious diseases Livestock Dairy farming The present study was conducted to assess the impact of associated risk factors and management practices on Babesia prevalence in cattles of district Toba Tek Singh, during 29-21. A total 68 (14.11%; CI=11.21-17.43) out of 482 cattle were recorded positive for Babesia infection. Babesia was recorded more prevalent during July-29 (25.64 %), whereas during December-29 and January-21 no case was recorded. The breed, age and gender of cattle represented significant differences (P<.5) for Babesia infection, whereas body condition of cattle s were found insignificant difference for associated risk factors. Among management practices, feeding, housing system, floor type and herd size were found significant differences (P<.5) for Babesia prevalence, while watering system found non-significant. 215 International Scientific Organization. All rights reserved. Capsule Summary: The feeding, housing, floor type and herd size had significant effect on Babesia prevalence in cattle. Cite This Article As: M. Iqbal, Tauseef-ur-Rehman, M.N. Khan, Z. Iqbal, M.S. Sajid, R.Z. Abbas, M.T. Javed, M.N. Abbas. Associated risk factors and management practices influence on Babesia prevalence in cattle of District Toba Tek Singh, Punjab, Pakistan. Current Science Perspectives 1(1) (215) 26-32. INTRODUCTION Livestock plays a key role in the economy of Pakistan. It contribute approximately 11.3% GDP during 28-9. Gross value addition of livestock at current cost factor has increased from PKR 152 billion (27-8) to PKR 1287 billion (28-9) shows an increase of 22.3 %. According to Livestock Census (26) in Pakistan, gross milk and meat production is 4.872 and 2.64 tons from domestic animals, respectively. The Babesiosis can lead to abortion, high mortality and lowered working efficiency of infected animals (Teglas et al., 25). Babesiosis is caused by intra-erythrocytic protozoan 26 parasites having worldwide distribution. The occurrence of the disease is dependent on the distribution of the vector ticks. The Babesia bovis and B. bigemina are the babesia species while B. bigemina is Boophilus decoloratus. Boophilus microplus are species of ticks which are responsible for the transmition of both B. bovis and B. bigemina in livestock. The mode of transmission between the ticks and the hosts is transovarial where either the nymph or the adults can transmit the disease (Young et al., 1988). The prevalence of protozoa, especially Babesia has been reported 37.1% from various parts of the world (Gonzales et al., 27; Klockiewicz et al., 27; Criado-Fornelio et al., 29). Babesia is more abundant in small ruminants

Iqbal et al. / Current Science Perspectives 1(1) (215) 26-32 (Theodoropoulos et al., 26; Yang et al., 29). Numerous studies are available on associated risk factors of protozoan infections in different countries viz., Iran (Razmi et al., 23), Kenya (Okuthe and Buyu, 26), Greece (Theodoropoulos et al., 26), Rwanda (Bazarusanga et al., 27), Bolivia (Gonzales et al., 27) and Uganda (Magona et al., 28). The age, sex, species, breed, season, feeding system and watering system are the major factors which are associated with the prevalence of protozoic fauna in domestic animals. In Pakistan, the Eimeria Theileria and Babesia genera are more prvelent in domestic animals in different regions of the country (Ahmad, 1992; Durrani et al., 25; Zahid et al., 25). Nasir et al., (29) the influence of specie, breed, age, sex on prevalence of Cryptosporidium in cattle and buffalos of district Lahore, Pakistan. In addition, seasonal variation in the prevalence of babesiosis has been studied in district Kasur by Zahid et al., (25). There is dearth of knowledge on the occurrence and determination of risk factors associated with babesiosis in Pakistan. Moreover, no studies studies was conducted to determine the associated risk factors and management practices effect on Babesia infections in cattle in central Punjab (District T. T. Singh), Pakistan. Therefore, the present study was designed to evaluate the prevalence, species composition, and associated risk factors with Babesia infections of cattles in district Toba Tek Singh (T.T. Singh), Pakistan. MATERIAL AND METHODS A cross-sectional study was carried out for epidemiological investigations on babesiosis from April, 29 to March, 21. 1. Age: Animals were divided into two age categories; adults (5 1 years) and young-stock (3 5 years). 2. Breed: Sahiwal, Friesian, Jersey and Cross-bred breeds were selected. 3. Sex: Both sexes were sampled during study. 4. Climate: Season-wise prevalence was noted separately as; (a) Cold season (December-March), (b) Hot season (April-June), (c) Rainy season (July-September) and (d) Postrainy season (October-November) 5. Husbandry and Management: Type of feeding system (ground feeding/trough feeding), housing system (open/closed), floor pattern (non-cemented/partially cemented), watering system (tap water/pond), herd size (larger (>2 animal)/smaller (<2 animal)) and body condition (good/poor) were also recorded during sampling. Collection of samples: A 5-1 ml blood collected from jugular vein using a sterile syringe in screw capped bottles containing EDTA as preservative and subsequently subjected to the analysis for Babesia infection identification was made by techniques given by Iqbal et al. (26). Meteorological data: The meteorological data i.e. month wise average temperature, relative humidity and rain fall was obtained from Meteorological Cell, Department of Crop Physiology, University of Agriculture, Faisalabad, Pakistan. Statistical analyses: Logistic analysis was performed using logit model including all variables in the model with backward elimination procedure. Factors with paired characteristics were analyzed using Odds Ratio (OR) and Mantel Haenszel (M. H.) chi-square. Hosmer Lemeshow goodness-of-fit test indicated that model fits well. All the analyses were carried out using SAS software package (1998) at 95% confidence level (SAS 1998). The prevalence was calculated for all data by using the following formula (Rehman et al., 211). Prevalence (%) = n/n x 1 Where, n and N are representing the number of infected individual and total number of animal sampled, respectively. Study area: Thepresent study was conducted in district T.T. Singh (3 33 to 31 2 N and 72 8 to 72 48 E) for the assessment of Babesia prevalence in cattle. It comprises of three tehsils (Gojra, Kamalia, and T.T. Singh) and 82 union councils (UC). It occupies an area of 3,252 km2 most of which is lowland, flooded during the rainy season. May, June and July are the hottest with maximum mean temperature (4.7 C) whereas December and January are the coldest months of the year with minimum mean temperature of 6 C of the area. An average annual rainfall is 254 381 mm (Rehman et al., 211). The map of Pakistan showing provinces and Districts including study area (Toba Tekh Singh) is shown in Fig. 1. Sampling units: The sampling was done randomly according to the number of primary units (UC s) and elementary units (animals) as reported by Thrusfield (28). The map grid method was used to Select primary units for collection of samples. Blood samples were collected randomly from 482 cattles irrespective of the cattles with no clinical symptoms. Samples were colected from 45 UC s out of 82 UC s. Development of questionnaire: The closed ended (dichotomous and multiple choice) questionnaire was made for the collection of information regarding associated risk factors (Thrusfield, 28) from farmers. Information regarding the following determinants was collected through questionnaire. 27 RESULTS The present study represented that the prevalence of Babesia among cattles depend on numerous risk factors and management practices. A total of 68 (14.11 %; CI=11.21-17.43) out of 482 studied cattles were recorded to have Babesia symptoms. Analysis of the risk factors by stepwise backward elimination logistic regression model (Table 1) and Mantel-Haenszel Chi-Square analysis (Table 2) revealed that age, sex, floor, herd size, housing and feeding system were correlated (P<.5) with Babesia prevalence. Babesia prevalence among breeds of cattles was recorded significantly different (P<.5).The body condition of cattles and watering system revealed non-significant difference (P>.5). The prevalence of babesia was recorded highest during the month of July 25.64% (CI=13.82-4.96). Whereas no symptoms of Babesia prevalence in cattles were recorded during December-29 and January-21 and increasing trends were recorded in the following months viz., February to onward. Month wise variation in the prevalence of Babesia (Fig. 1).

Prevalence (%) Iqbal et al. / Current Science Perspectives 1(1) (215) 26-32 Table 1: Logistic regression analysis of associated factors with risk of Babesia infection in cattle Term Odds ratio 95% C. I. P-value Breed.375.272-.517. Age 3.522 1.759-7.51. Feeding system 2.643 1.387-5.36.3 Fig. 1: Map of Pakistan provinces and Punjab showing Districts including Toba Tekh Singh (T. T. Singh) area under study. Floor.218 Watering system 3.835.11-.47 1.924-7.646.. 3 25 2 15 1 5 Apr-9 May The weather conditions have major impact on the prevalence of babesia and the highest prevalence in cattles were recorded in summer season (July and August).. The prevalence of Babesia was recorded % during Dec-29 and Jan-21 and rain fall was recorded minimum during these months. The impact of rain fall, temperature, relative humidity on Babesia prevalence in cattles (Fig. 3). Associated risk factors June July August September October November December Jan-1 February March Fig. 2: Month wise prevalence of Babesia in cattle form T. T. Singh District, Punjab, Pakistan Numerous associated risk factors were evaluated such as The cattle breeds viz., the Fresian (32.18%; 28/87; χ2=42.422; OR=.375) was recorded to be more affected followed by Jersey (2.21%; 19/94) and Cross-bred (12.38%; 13/15). While indigenous breed (Sahiwal) was recorded at least risk of babesiosis with the prevalence of only 4.8% (8/196). Young and adult cattles revealed significant difference (P<.5). The sex (male and female) revealed significant differences (P<.5), where females cattles (19.78%; 54/273; χ2=16.683; Housing system 9.17 OR=.271) were recorded more susceptible for babesia than male cattles (6.7%; 14/29). Body conditions of the cattles represented non-significant difference (P>.5) Table 2 and Fig. 4. Management risk factors The management practices plays a significant role in the prevalence of Babesia like feeding system, the ground fed cattles were recorded more susceptible for Babesia infection trough fed cattles. A total of 41 (2.29%; χ2=1.969; OR=2.643) ground fed cattles out of 22 of cattles were recorded infected with with Babesia infection whereas just 9.64% (27/28) was recorded in cattle fed in trough. The housing system revealed significant difference (P<.5) where a total of 2.2% (4/198; χ2=1.278; OR=2.643) cattles kept under closed system were recored infected with babesia while low prevelance was recorded in open housing system (9.86%; 28/284). Floor type revealed significant difference (P<.5) for the Babesia prevalence. Babesia Prevalence was recorded high 18.82% (51/271; χ2=11.316; OR=.218) in cattles kept in kaccha floor (non-cemented), while it was 8.6% (17/211) in partially cemented floor. The herd size was also found a determinant factor for Babesia infection. In farms of cattle herd size >2, 19.77% (35/196; χ2=7.396) animal found affacted, while 35 out of 35 (11.48%) cattle in herd size <2 were found positive for Babesia ( Table 2; Fig. 5). DISCUSSION 4.144-2.16. HOSMER-LEMESHOW GOODNESS-OF-FIT TEST: P =.752 28

Ground Trough Noncemented Partially cemented Open Close Tap water Pond >2 <2 Poor Good Animals infected and Examine Prevalence (%) Animals infected and Examine Prevalence (%) Young Adult Male Female Cross bred Shaiwal Fresion Jersey Prevalence (%), Temp (ºC), Relative Humidity (%) Rain Fall (mm) Iqbal et al. / Current Science Perspectives 1(1) (215) 26-32 9 8 7 6 5 4 3 2 1 Temperature R. humidity Rain fall Prevalence (%) Apr-9 May June July August September October November December Jan-1 February March Fig. 3: Association of weather condition with risk of Babesia in cattle from T. T. Singh District, Punjab, Pakistan 3 25 2 15 1 5 Animals infected Animals Examined Prevalence (%) Age Sex Breed Fig 4: Association of associated risk factor (age, sex and breed) with risk of Babesia in cattle from T. T. Singh District, Punjab, pakistan 35 3 25 2 15 1 5 Animals infected Animals examined Prevalence (%) Feeding system Floor Housing system Watering Herd size Body condition Fig 5: Association of management risk factor (feeding system, floor type, housing system, watering system, herd size and body condition) with risk of Babesia in cattle from T. T. Singh District, Punjab, Pakistan In the present study a total of 482 cattles of four breed, including adults and young of both gender were studied. The overall prevalence of Babesia was observed 14.11% (68/482) 29 14 12 1 8 6 4 2 35 3 25 2 15 1 5 6 5 4 3 2 1 including all risk factors and management practices. Results revealed that associated risk factors, management practices and weather condition had strong correlation with Babesia prevalence in cattle in District T. T. Singh,.Previously associated risk factors with babesiosis and prevalence of Babesia species in small ruminants (Iqbal et al., 211) while Zulfiqar et al.(212) hematological and serum biochemical profile in large ruminants were studied in southern Punjab and revealed similar results. Such studies are also available on other animals like Ahmad et al. (27 and 211) reported the Babesia prevalence in dogs and cats, respectively form Lahore, Pakistan and associated it with various risk and management factors. Rashid et al (21) reported the prevalence of babesiosis among Lohi sheep in the Livestock Experiment Station, Qadirabad, and District Sahiwal. Furthermore, the results of Babesia prevalence and associated risk factors study also correlate with with numerous reports from countries over the world such as Egypt (Mazyad et al., 22), Sri Lanka (Jorgensen et al., 1992), Gambia (Mattioli et al., 1997), Spain (Almeria et al., 21; Nagore et al., 24), Portugal (Criado-Fornelio et al., 23), Pakistan (Khan et al., 24), USA (Teglas et al., 25), Bangladesh (Chowdhury et al., 26), Kenya (Okuthe and Buyu, 26), Turkey (Altay et al., 28), China (Guan et al., 28), Iran (Razmi et al., 23), Greece (Papadopoulos et al., 1996), Israel (Yeruham et al., 1992), Zambia (Nalubamba et al., 211) and Sudan (Salih et al., 28), they reported the Babesia prevalence ranging from 1-1% in different environment and risk factors. However, our findings vary from Papadopoulos et al. (1996), Karatepe et al. (23) and Hosein et al. (27), they reported 52.1%, 23.63% and 5.92% bovine babesiosis, respectively versus present study (14.11 %). This variation might be due to the application of highly sensitive tests like indirect fluorescent antibody technique for diagnostic purpose. In contrast to above reports Mazyad and Khalaf (22) reported a significantly lower Babesia prevalence i.e. only 2.73% among 475 sheep, which may be due to low existence of vector parasites which is responsible for the spread of disease or may be due to more variation in species as well as different agroclimatic condition. In the present study, the adults and female cattles were recorded more susceptible to Babesia infection. Previously Iqbal et al. (211) and Zulfiqar et al. (212) reported significant difference in the Babesia prevalence in different age and sex groups of of cattles. The different breed of cattles are not equally susceptible for

Iqbal et al. / Current Science Perspectives 1(1) (215) 26-32 Table 2: Mantel-Haenszel Chi-Square analysis of all hypothesized risk factors with Babesia infection in cattle Mantel-Haenszel Chi- Associated determinants Variables Prevalence Square (P-value) Age Sex Young 6.17% (14/227) 22.2797 Adult 21.18% (54/255) (<.1) Male 6.7% (14/29) 16.6828 Female 19.78% (54/273) (<.1) Breed Cross bred 12.38% (13/15) Sahiwal 4.8% (8/196) Fresion 32.18% (28/87) Jersey 2.21% (19/94) 42.4218 (<.1) Feeding system Floor Housing system Watering system Herd size Body condition Ground 2.29% (41/22) 1.9694 Trough 9.64% (27/28) (.9) Kacha 18.82% (51/271) 11.3162 Partially cemented 8.6% (17/211) (.8) Open 9.86% (28/284) 1.2778 Close 2.2% (4/198) (.13) Tap water 11.65% (29/249) 2.5698 Pond 16.74% (39/233) (.189) >2 19.77% (35/177) 7.3956 <2 11.48% (35/35) (.65) Poor 15.38% (34/221).5479 Good 13.3% (34/261) (.4592) Babesia infection (Bashir et al., 29; Muhanguzi et al., 21; Ahmad et al., 211 present study). The weather conditions vary throughout the year e.g. hot dry to humid rainy seasons. rainy season was recorded the most favourable season for the Babesia prevalence (25.64 %, July-29), while least favourable is winter season ( %, Dec- 29 and Jan-21). Ahmad et al. (211) reported a significant variation in Babesia prevalence during different seasons and highest prevalence recorded during summer and autumn. Birkenheuer et al. (1999) and Kar et. al. (28), they reported that higher Babesia prevalence with high risk of vectors during peak infection periods. The highest Babesia prevalence during rainy season may be due to the rainy season provide favourable environment for the potential growth of parasites. The management practices were recorded playing a key role in prevalence of Babesia except watering system. The poor hygienic situations and overcrowded conditions are the major cause of babesia infection in non-cemented floor, closed 3 housing system and large herds due to greater contamination (Soulsby, 26; Khan et al., 211). Iqbal et al. (211) reported that the management risk factors are significantly important for infections and multiplication of parasites furthermore larger, mixed herds, dog (having ticks on their body present in herd) are among the major risk factors for the spread of babesiosis in animals. Khan et al. (211) reported that high stock increases the environmental contamination with oocysts and therefore the risk of an infection and prevalence might be more common in animals confined in small and contaminated area. Radostits (29) and Rehman et al. (211) gave the precaution measure regarding feeding system, housing, floor type and watering system which are more suitable to control Babeiosis. They also suggested keep animal separate form other species and in small herd size. REFERENCES

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