Prevalence of trypanosomiasis caused by Trypanosoma brucei (Kinetoplastea, Trypanosomatidae) in domestic ruminants from Southern Punjab, Pakistan

Background and Aim: Trypanosomiasis, a parasitic infection caused by various Trypanosoma species, poses a significant threat to global livestock, affecting both human health and economic sectors. This study aimed to estimate the prevalence of trypanosomiasis in Southern Punjab, Pakistan, focusing on key ruminant species, including camels, cattle, buffaloes, goats, and sheep. Materials and Methods: A total of 240 blood samples, comprising 48 samples from each animal species (camel, cattle, buffaloes, goat, and sheep) were collected from three districts in Southern Punjab. The collected samples were subjected to thin smear microscopy, DNA extraction, and polymerase chain reaction (PCR) amplification. The molecular characterization was conducted using the TBR primer set, which targeted repeated satellite DNA regions and the cytochrome oxidase II gene of Trypanosoma brucei. Results: About 22.08% (53/240) of overall samples were positive for trypanosomiasis, with prevalence rates being 23.75% (19/80), 21.25% (17/80), and 21.75% (17/80) for districts Muzaffargarh, Lodhran, and Bahawalpur, respectively. 5.83% (14/240) of samples tested for T. brucei using PCR were positive in the districts of Muzaffargarh 7.50% (6/80), Lodhran 5.00% (4/80), and Bahawalpur 5.00% (4/80). Among the animals tested, camels had the highest positivity rate. The microscopic examination confirmed infection rates of 45.83% (22/48) for camels, 18.75% (9/48) for cattle, 8.33% (4/48) for buffaloes, 18.75% (9/48) for goats, and 18.75% (9/48) for sheep (p < 0.001). PCR results did not reveal substantial differences (p < 0.05) in prevalence: camels 12.50% (6/48), cattle 6.25% (3/48), buffaloes 0% (0/48), goats 8.33% (4/48), sheep 2.08% (1/48); while distinct disparities were detected district-wise: Muzaffargarh 23.75% (19/80), Lodhran 21.25% (17/80), and Bahawalpur 21.25% (17/80). The PCR results for these districts were insignificantly different: 7.50% (6/80), 5% (4/80), and 5% (4/80). The microscopic infection rate in camels from Bahawalpur was 56.30% (9/16). The microscopic analysis in Buffaloes reported a 6.30% (1/16) infection rate, but PCR results indicated no infections (0%) in any district. A significant difference (p < 0.001) in identifying Trypanosoma species was found between positively and negatively tested animals in both microscopic and PCR methods. Conclusion: This study emphasizes the necessity of regularly using PCR-based screening for its superior sensitivity and specificity over traditional microscopy. The varying occurrence of trypanosomiasis among districts reflects the intricate nature of this diseases epidemiology in the region. Reducing economic losses from trypanosomiasis in Southern Punjab, Pakistan, requires targeted interventions, such as vector control measures and farmer education. Keywords: domestic ruminants, microscopic examination, Pakistan, polymerase chain reaction detection, Trypanosoma brucei, trypanosomiasis.