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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.

Parsley (Petroselinum crispum) is herb with many biological and medicinal benefits for humans. However, growth on zinc (Zn) and cadmium (Cd) contaminated sites might get severely affected due to over accumulation of heavy metals (HM) in different plant tissues. Antioxidants play a crucial role in minimizing the negative effects of HM. The present study investigates the effects of Zn and Cd stress on P. crispum morphological parameters, enzymatic/non-enzymatic antioxidant profiling and metal accumulation in shoot/root. Plants were exposed to different concentrations of Zn (50, 100, 150 and 200 µM) and Cd (10, 20, 40 and 80 µM) along with control (no stress), in soil-less Hoagland's solution. The results showed that Zn and Cd substantially decrease the growth parameters with increased contents of malondialdehyde (MDA), hydrogen peroxide (H2O2), and electrolyte leakage (EL). Non-enzymatic antioxidant activities, like total phenolic contents (TPC) and ferric reducing antioxidant power (FRAP), were induced high in leaves only upon Cd stress and contrarily decreased upon Zn stress. Total flavonoid contents (TFC) were decreased under Zn and Cd stress. Enzymatic antioxidant activities like superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were also strongly induced upon Cd stress. At the same time, SOD and guaiacol peroxidase (GPX) activity was induced significantly upon Zn stress. Cd uptake and accumulation was notably high in roots as compared to shoots, which suggests P. crispum have a reduced ability to translocate Cd towards aboveground parts (leaves). Additionally, strong induction of antioxidants by P. crispum under Cd stress might indicate the capacity to effectively re-modulate its physiological response. However, further investigations regarding other HMs and experiments at the molecular level are still needed.