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Pakistan has a huge sheep population (37.2 million in 2024) that is largely unexplored for the presence of vector transmitted parasites. Present study was aimed to document the prevalence of Anaplasma sp. , Anaplasma ovis , Theileria ovis and Theileria lestoquardi in sheep blood samples (N = 329) that were collected from six districts (Muzaffargarh, Rajanpur, Dera Ghazi Khan, Layyah, Taunsa and Khanewal) during August till December 2024 and to report the genetic diversity of screened pathogens. Molecular analyses revealed that the prevalence of Anaplasma sp. , Anaplasma ovis and Theileria ovis in screened sheep was 11%, 20% and 21% respectively. None of the screened sheep was Theileria lestoquardi infected. Co-infection of the screened pathogens was also observed. Presence of the detected pathogens was confirmed by DNA sequencing and subsequent BLAST analysis. Phylogenetic analysis revealed that these pathogens displayed genetic similarities with the sequences that were deposited from various countries across the globe. Prevalence of all screened pathogens varied significantly between the sampling districts. Similarly, the Anaplasma sp., Anaplasma ovis and Theileria ovis prevalence varied significantly among the sheep breeds. Anaplasma ovis infection was more common in large herds and in un-infested sheep. Theileria ovis infection was more frequent in small herds. In conclusion, we are reporting the presence of Anaplasma sp., Anaplasma ovis and Theileria ovis in Pakistani sheep that were enrolled from all six districts. Large-scale studies are recommended in various geo-climatic regions of Pakistan to confirm the genetic diversity, epidemiology and host-pathogen interactions that will contribute towards effective control of these infections among the local sheep population.

Heat stress (HS) is considered a severe concern for rabbit buck’s health, welfare and reproductive features. Thymoquinone (THQ) is a natural molecule with robust antioxidant, antimicrobial, and anti-inflammatory properties. Thereafter, three months of feeding research was applied to assess the role of THQ-enriched diets in improving the health, semen attributes and testicular changes of stressed rabbit bucks. Forty rabbit bucks were used in this research, and animals were fed diets containing 0 (THQ0; serve as a heat stress group), 100 (THQ100), 200 (THQ200), and 400 (THQ400) mg/kg of THQ for three months continually during the summer season. Outcomes displayed that THQ significantly enhanced FBW (final body weight), DBWG (daily body weight gain), and FI (feed intake) but did not affect FCR (feed conversion ratio). Compared with the THQ0 group, dietary THQ addition significantly decreased creatinine, TB (total bilirubin), TG (triglycerides), and GGT (gamma-glutamyl transferase) levels. Bucks receiving 200 mg of THQ had higher TAC (total antioxidant capacity) and GSH (glutathione) than other groups. At the same time, MDA was significantly shrunk in THQ-treated groups relative to the THQ group. The levels of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were significantly diminished (P <0.001), but nitric oxide and lysosome activity were significantly increased in bucks given diets containing THQ (200 and 400 mg/kg) as relative to the TH0 group. Libido score and reaction time were significantly improved by THQ dietary inclusion. Semen attributes, including volume, motility, concentration, and vitality, were significantly augmented in THQ100 and THQ200 groups. Histological screening of testicular tissues showed moderate to severe degenerative and necrotic changes in the testicular tissues of bucks exposed to HS. This was enhanced with the supplementation of THQ at different levels. Collectively, THQ (200 or 400 mg/kg diet) can improve health, semen attributes and restore testicular damage in rabbit bucks via boosting antioxidant and reducing inflammatory cytokines.