Epidemiological and haemato-biochemical study on bovine babesiosis in and around Bidar district, Karnataka

Introduction

Bovine babesiosis, a tick-borne disease caused by intraerythrocytic protozoa of the genus Babesia (phylum Apicomplexa), results in significant economic losses globally, affecting both domestic and wild bovids (Almazan et al., 2022) [1] . The key etiological agents include Babesia bovis, Babesia bigemina, Babesia divergens and Babesia major (Santos et al., 2023) [33] . Babesia bigemina ranks first among the top ten economically important parasitic diseases of domestic cattle in India (Minjauw and McLeod, 2003) [25] . The clinical manifestations of an acute babesiosis generally run a course of three to seven days. Fever (>40 °C) is usually present for several days before other clinical signs become apparent, followed by inappetence, depression, increased respiratory rate, weakness, and reluctance to move. Haemoglobinuria is an overt clinical sign, hence the disease is known as red water. Anaemia and jaundice develop especially in protracted cases, while muscle wasting, tremors, and recumbency develop in advanced stages followed terminally by coma (DeVos and Potgieter, 1994) [9] . Keeping in view the significance of bovine babesiosis, the present study was conducted to evaluate the prevalence and haemato-biochemical parameters in cattle affected with babesiosis.

Blood samples were collected from cattle presented to the VCC and peripheral Veterinary clinics exhibiting clinical symptoms suggestive of haemoprotozoan infections, namely pyrexia, haemoglobinuria, pale or icteric mucous membranes, and severe tick infestation. Of these, 24 clinical cases of bovine babesiosis confirmed by blood smear microscopy were selected for intensive clinical, haematological and biochemical tracking. Blood sampling was performed via aseptic jugular vein puncture into three sets of tubes: (1) EDTA tubes (2 mL) processed for peripheral thin blood films, automated haematology validation, and PCR studies; and (2) clot activator tubes (4 mL) allowed to clot for serum separation to facilitate semi-automated biochemistry profiling. Haematological parameters were quantified using the Merilyzer CelQuant 360 VET fully automated blood cell counter to determine haemoglobin (Hb), total erythrocyte count (TEC), packed cell volume (PCV), total leukocyte count (TLC), and total platelet count (PLT). Manual differential leukocyte count (DLC) was carried out under oil immersion (100×) on Giemsa-stained thin blood smears. Serum biochemical indices including total protein, albumin, globulin, blood urea nitrogen (BUN), creatinine, total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were processed on a semi-automated biochemical analyser (Microlab 300, Eli Tech Group) using ERBA diagnostic reagent kits.

Results

Out of 954 cattle presented to the medicine ward over the three-year timeline, 13 were definitively confirmed positive for babesiosis, yielding an overall prevalence of 1.36% (Table 1). Year-wise distribution indicated a peak point prevalence during 2022-23 (46.15%). Risk factor stratification highlighted a distinct gender predisposition, with female cattle manifesting a higher point prevalence (53.85%) than males (46.15%), a variation strongly correlated with the physiological stresses of lactation and reproduction. Age-wise distribution showed the highest infection rates in cattle under 2 years of age (46.15%), followed by the 2-6 years cohort (30.77%), reflecting relative immunological immaturity in young stock. Breed evaluation revealed that non-descript indigenous cattle comprised 69.23% of cases, while crossbred Holstein Friesian (HF) cattle comprised 30.77% (Table 1). Seasonally, a major surge occurred during the monsoon/rainy season (61.54%).

Discussion 4.1 Prevalence

The overall prevalence of bovine babesiosis in the present study was found to be 1.36%. Similarly, lower prevalence rates of 1.56% were observed by Singh et al. (2012) [37] and Sharma et al. (2016) [34] . Kumar et al. (2016) [20] reported a prevalence of 54.0%; Guswanto et al. (2017) [14] noted 27.1% seroprevalence; Siddique et al. (2020) [36] reported 26.86%; Haidy et al. (2023) [15] reported 13.88%; and Aziz et al. (2025) [3] reported a prevalence of 14.5%. The lower prevalence in the present study may be attributable to the assessment of a smaller number of samples presented to VCC, Veterinary College, Bidar. The gender-wise point prevalence study revealed a higher prevalence in female cattle (53.84%) compared to male cattle (46.15%), in accordance with Hamsho et al. (2015) [16] , Mahmud et al. (2015) [23] , Zia and Nazir (2019) [42] , and Mushahary et al. (2020) [27] . This difference is strongly correlated with the physiological stress of reproduction and lactation. However, Casa et al. (2025) noted maximum cases in male cattle, and Aziz et al. (2025) [3] highlighted the importance of considering local geographical factors, specific parasite species, and management systems while assessing disease risk. The age-wise point prevalence study revealed the highest prevalence in animals under 2 years, in accordance with Mahmud et al. (2015) [23] , Hussain et al. (2017) [18] , Bary et al. (2018) [6] , and Aziz et al. (2025) [3] , while Siddique et al.

(2020) [36] reported higher prevalence in adult cattle. The highest prevalence in young animals may be attributed to immunological immaturity.

The breed-wise prevalence study revealed that non-descript cattle comprised the majority of cases (69.23%), while crossbred HF cattle comprised 30.77%, yielding a breedspecific prevalence of 14.33% in crossbreds. Higher susceptibility of exotic/crossbred cattle is largely attributable to their lack of natural resistance to tropical ticks and pathogens when introduced into endemic areas (Hussain et al., 2017; Bary et al., 2018; Mushahary et al., 2020) [18,6,27] . Anbu et al. (2020) [2] reported higher prevalence (5.55%) in non-descript animals exposed to high tick challenge environments, which can override genetic resistance.

The season-wise prevalence study revealed the highest prevalence during the rainy season, in agreement with Bhatnagar et al. (2015) [7] , Parmar and Upadhyay (2017) [30] ,

Barman et al. (2018) [5] , and Kumar et al. (2018) [21] . High humidity and moderate-to-high temperatures during this period are ideal for tick hatching, development, and questing behaviour. Cold temperatures in winter slow tick metabolic activity and prolong the Babesia life cycle, leading to a significant drop in disease transmission (Bhatnagar et al., 2015; Kumar et al., 2018) [7,21] .

Haematological findings

Values of haemoglobin, total erythrocyte count, and packed cell volume were significantly decreased in babesiosisaffected cattle compared to healthy controls, consistent with anaemia. These findings accord with Mervat et al. (2013) [24] , Mahmoud et al. (2015) [22] , Tufani et al. (2015) [40] , El-Hamed et al. (2016) [11] , Aziz et al. (2020) [4] , and Charaya et al. (2021). In babesiosis, major alteration in total RBC osmotic fragility causes even intact unaffected red cells to become easily destroyed, creating haemolysis (Kakoma and Ristic, 1984) [19] . No significant differences were observed in MCV, MCH, and MCHC values in babesiosis-affected cattle compared to the healthy control group, in accordance with El-Hamed et al. (2016) [11] and Ganguly et al. (2017) [13] .

There was a significant decrease (p ≤ 0.05) in platelet values in babesiosis-affected cattle. Similar findings were reported by Tufani et al. (2015) [40] , Noha et al. (2016) [29] , Hashem et al. (2018) [17] , Haidy et al. (2023) [15] , and Roopali et al.

(2025) [31] . The thrombocytopenia observed may be attributed to babesiosis-associated immune thrombocytopenia (Narurkar et al., 2017) [28] . There was a significant decrease (p ≤ 0.05) in total leukocyte count in babesiosis-affected cattle, similar to findings of El-Hamed et al. (2016) [11] , Noha et al. (2016) [29] , Sharma et al. (2016) [34] , and Hashem et al. (2018) [17] . No significant differences were observed in differential leukocyte parameters (neutrophils, lymphocytes, monocytes, eosinophils, and basophils).

Biochemical findings

There was a significant decrease in total protein and globulin levels in cattle presenting with babesiosis, as reported by El-Hamed et al. (2016) [11] , Noha et al. (2016) [29] , Hashem et al. (2018) [17] , Aziz et al. (2020) [4] , and Roopali et al. (2025) [31] , attributable to the harmful effect of toxic metabolites of Babesia species on liver cells (Mohamed Saied, 2017) [26] . Albumin showed no significant difference, in accordance with Shinde et al. (2019) [35] and Aziz et al. (2020) [4] . The A:G ratio was significantly elevated, reflecting complex factors including severe liver dysfunction, dehydration, or co-infections. Significant elevation in BUN and creatinine was observed, consistent with El-Hamed et al. (2016) [11] , Ganguly et al. (2017) [13] , Sivajothi et al. (2024) [38] , and Roopali et al. (2025) [31] . Harmful effects of the parasite's toxic metabolites on kidney cells lead to azotaemia. Degraded byproducts of damaged RBCs deposited in the kidneys reduce creatinine excretion (Talkhan et al., 2010) [39] . Significant elevation in total bilirubin was observed, consistent with multiple prior reports ( [11,10,4,38] . The increase in total bilirubin results from major alteration in total RBC osmotic fragility causing haemolysis (Kakoma et al., 1984) [19] . Significantly increased values of AST, ALT, and ALP were observed in the present investigation, consistent with Samad et al. (2011) [32] , El-Hamed et al. (2016) [11] , Sharma et al. (2016) [34] , Hashem et al. (2018) [17] , Haidy et al. (2023) [15] , Sivajothi et al. (2024) [38] , and Roopali et al. (2025) [31] . The harmful effect of toxic metabolites of Babesia spp. on hepatocytes leads to cellular necrosis and elevated serum transaminase and ALP activities (Yeruham et al., 2003; Esmaeilnejad et al., 2021) [41,12] .

Conclusion

The present study confirms the endemic nature of bovine babesiosis in and around Bidar district, Karnataka. Prevalence was highest in female cattle and in the age group of less than 2 years, followed by 2 to 6 years and above 6 years. Prevalence was higher in non-descript cattle and was highest during the rainy season. Haematological investigation showed significant decrease in haemoglobin, total erythrocyte count, packed cell volume, platelets, and total leukocyte count. Biochemical parameters showed significant increase in A:G ratio, BUN, creatinine, total bilirubin, AST, ALT, and ALP, with significant decrease in total protein and globulin in affected cattle.

Disclaimer (Artificial Intelligence)

Author(s) hereby declare that NO generative AI technologies such as Large Language Models (ChatGPT, COPILOT, etc.) and text-to-image generators have been used during the writing of this manuscript.

References

1. Almazan C, Scimeca RC, Reichard MV, Mosqueda J. Babesiosis and theileriosis in North America. Pathogens. 2022;11(2):168.
2. Anbu RKR, Mangalanathan VB, Ganapathy S, Alagesan SK. Epidemiological studies on bovine tickborne haemoparasitic diseases in Chennai. Int J Livest Res. 2020;10(2):36-45.
3. Aziz KJ, Waly TA, Shekhmhamad OQ. Molecular epidemiology of babesiosis in dairy cattle in Erbil Province, Iraq. Passer J Basic Appl Sci. 2025;7(1):2936.
4. Aziz PR, Marodia S, Ganesan PI, Sharma CS. A clinical study on hemato-biochemical changes in cows affected with babesiosis. J Pharm Innov. 2020;9(2):242245.
5. Barman U, Dutta TC, Baishya BC, Goswami S, Islam S, Kakati P. Study of prevalence and diagnosis of babesiosis in cattle. J Entomol Zool Stud. 2018;6(3):1613-1616.
6. Bary MA, Ali MZ, Chowdhury S, Mannan A, Moula MM, Bhuiyan ZA, et al. Prevalence and molecular identification of haemoprotozoan diseases of cattle in Bangladesh. Adv Anim Vet Sci. 2018;6(4):176-182.
7. Bhatnagar CS, Bhardawaj B, Sharma DK, Meena SK. Incidence of haemoprotozoan diseases in cattle in Southern Rajasthan, India. Int J Curr Microbiol Sci. 2015;4(3):509-514.
8. Charaya G, Goel P, Kaur J, Kumar M, Singh Y. Alterations in haemato-biochemical parameters in crossbred cattle suffering from babesiosis. Indian Vet Med J. 2021;41:65-67.
9. DeVos AJ, Potgieter FT. Infectious Diseases of Livestock. Cape Town: Oxford University Press; 1994. p. 278-294.
10. El-Diasty MM, Rasheed NM, Kheder Z, El-Maged RR. Biochemical, pathological and molecular studies on babesiosis in calves. Egypt J Agric Res. 2017;95(3):1269-1283.
11. El-Hamed HAA, Salem SM, Ibrahim HN. Haematobiochemical alterations in cattle suffering from anaemia and their effect on quality of some meat. Egypt J Chem Environ Health. 2016;2(2):232-249.
12. Esmaeilnejad B, Dalir-Naghadeh B, Tavassoli M, AsriRezaei S, Mahmoudi S, Rajabi S, et al. Assessment of hepatic oxidative damage, paraoxonase-1 activity, and lipid profile in cattle naturally infected with Babesia bigemina. Trop Anim Health Prod. 2021;53(2):219.
13. Ganguly A, Bisla RS, Ganguly I, Singh H, Bhanot V, Chaudhri SS. Direct blood PCR detection of Babesia bigemina and its effect on haematological and biochemical profile in crossbred cattle of eastern Haryana. Indian J Anim Res. 2017;51(1):141-145.
14. Guswanto A, Allamanda P, Mariamah ES, Munkjargal T, Tuvshintulga B, Takemae H, et al. Evaluation of immunochromatographic test strips for the serological detection of Babesia bovis and Babesia bigemina infection in cattle from Western Java, Indonesia. Vet Parasitol. 2017;239:76-79.
15. Haidy EM, Hala NI, Nahla HS. Molecular characterization, hematobiochemical changes and therapeutic management of tick-borne haemoparasites in naturally infected cattle. J Adv Vet Res. 2023;13(10):2039-2046.
16. Hamsho A, Tesfamarym G, Megersa G, Megersa M. A cross-sectional study of bovine babesiosis in Teltele district, Borena zone, Southern Ethiopia. J Vet Sci Technol. 2015;6(3):1-4.
17. Hashem MA, Neamat-Allah AN, Gheith MA. A study on bovine babesiosis and treatment with reference to hematobiochemical and molecular diagnosis. Slov Vet Res. 2018;55(20):165-173.
18. Hussain S, Ashraf K, Anwar N, Jamal MA, Naeem H, Ahmad N, et al. Diagnosis of Babesia bovis infection in indigenous and crossbred cattle with comparison between conventional and molecular diagnostic techniques. J Infect Mol Biol. 2017;5(1):1-6.
19. Measures. Dordrecht: Springer Netherlands; 1984. p. 85-93.
20. Kumar B, Maharana BR, Prasad A, Joseph JP, Patel B, Patel JS. Seasonal incidence of parasitic diseases in bovines of south western Gujarat (Junagadh), India. J Parasit Dis. 2016;40(4):1342-1346.
21. Kumar B, Mondal DB, Jithin MV. Prevalence of babesiosis in cattle in Patna region, India. Int J Curr Microbiol Appl Sci. 2018;7:5167-5174.
22. Mahmoud MS, Kandil OM, Nasr SM, Hendawy SH, Habeeb SM, Mabrouk DM, et al. Serological and molecular diagnostic surveys combined with examining haematological profiles suggests increased levels of infection and haematological response of cattle to babesiosis infections compared to native buffaloes in Egypt. Parasit Vectors. 2015;8(1):319.
23. Mahmud MMA, Belal SMSH, Hossain MA. Prevalence of theileriosis and babesiosis in cattle in Sirajganj district of Bangladesh. Res Agric Livest Fish. 2015;2(1):79-86.
24. Mervat EI, Radwan, Omnea Abdel Hamied, Abdel Fatah Ali. Evaluation of erythrocytes antioxidant mechanisms in bovine babesiosis and current advances treatment in Kaliobea Governorate. Am J Infect Dis Microbiol. 2013;1(4):59-63.
25. Minjauw B, McLeod A. Tick-Borne Diseases and Poverty. Research Report. Centre for Tropical Veterinary Medicine, University of Edinburgh; 2003. p. 1-124.
26. Mohamed Saied MM. Studies of naturally infected babesiosis and its effect on some hematological and biochemical parameters in cattle in Qena, Egypt. Dairy Vet Sci J. 2017;4(3):555640.
27. Mushahary D, Bhattacharjee K, Sarmah PC, Deka DK, Kakati P. Quantification of Babesia bigemina from cattle in Indo-Bhutan border district of Assam by realtime PCR. J Entomol Zool Stud. 2020;8(6):281-284.
28. Narurkar R, Mamorska-Dyga A, Agarwal A, Nelson JC, Liu D. Babesiosis-associated immune thrombocytopenia. Stem Cell Investig. 2017;4:1.
29. Noha YS, Shimaa GY, Heba SF, Mohammed AE. Clinical, hemato-biochemical alterations and oxidantantioxidant biomarkers in Babesia-infected calves. Int J Vet Sci. 2016;4(1):17-22.
30. Parmar T, Upadhyay AK. Prevalence of haemoprotozoan diseases of large animals in Kumaon Tarai of Uttarakhand. Int J Recent Sci Res. 2017;8(6):17583-17584.
31. Roopali B, Roy M, Pavitra, Jayashree, Basavanand, Manoj Kumar, et al. Successful therapeutic management of babesiosis in a calf. Uttar Pradesh J Zool. 2025;46(10):240-244.
32. Samad L, Rahmani M, Mohri M, Madadgar O. Changes in serum iron concentration and hepatic enzyme activities in cattle infected with Theileria annulata and Babesia bigemina. Comp Clin Pathol. 2011;21:829832.
33. Santos JHM, Siddle HV, Raza A, Stanisic DI, Good MF, Tabor AE. Exploring the landscape of Babesia bovis vaccines: Progress, challenges, and opportunities. Parasit Vectors. 2023;16(1):274.
34. Sharma A, Singla LD, Batth AK, Kaur P. Clinicopathobiochemical alterations associated with subclinical babesiosis in dairy animals. J Arthropod Borne Dis. 2016;10(2):258-266.
35. Shinde RM, Bhikane AU, Naraladkar BW, Masare PS. Antioxidants as an adjunct therapy in clinical management of babesiosis in cattle: A novel approach. Rumin Sci. 2019;8(1):93-100.
36. Siddique RM, Sajid MS, Iqbal Z, Saqib M. Association of different risk factors with the prevalence of babesiosis in cattle and buffalo. Pak J Agric Sci. 2020;57(2):517-524.
37. Singh NK, Singh H, Jyoti, Haque M, Rath SS. Prevalence of parasitic infections in cattle of Ludhiana district, Punjab. J Parasit Dis. 2012;36(2):256-259.
38. Sivajothi S, Dhanamjayam P, Sudhakara B, Shobhamani B. Serum biochemical profile alterations in cattle with clinical babesiosis. Chem Sci Rev Lett. 2024;11(41):120-124.
39. Talkhan OFA, Radwan MEI, Ali MA. Cattle babesiosis and associated biochemical alteration in Kalubyia Governorate. Nat Sci. 2010;12:24-27.
40. Tufani NA, Fazili MR, Malik HU, Beigh SA, Dar KH. Clinico-haematological profile and therapeutic management of acute babesiosis in a Holstein-Friesian crossbred cow. Vet Clin Sci. 2015;3(3):11-14.
41. Yeruham I, Avidar Y, Aroch I, Hadani A. Intra-uterine infection with Babesia bovis in a 2-day-old calf. J Vet Med. 2003;50(2):60-62.
42. Zia A, Nazir Q. Investigation on prevalence of babesiosis in various areas of Taunsa Sharif district Dera Ghazi Khan, Punjab, Pakistan. Buffalo Bull. 2019;38(3):485-489.