(1) Background: Surra is a debilitating disease of wild and domestic animals caused by Trypanosoma evansi (T. evansi), resulting in significant mortality and production losses in the affected animals. This study is the first to assess the genetic relationships of T. evansi in naturally affected buffaloes from Multan district, Pakistan, using ITS-1 primers and evaluating the effects of parasitemia and oxidative stress on DNA damage and hematobiochemical changes in infected buffaloes. (2) Methods: Blood samples were collected from 167 buffaloes using a multi-stage cluster sampling strategy, and trypomastigote identification was performed through microscopy and PCR targeting RoTat 1.2 and ITS-1 primers. Molecular characterization involved ITS-1 via neighbor-joining analysis. The impact of parasitemia loads was correlated with oxidative stress markers, genotoxicity, and hematobiochemical parameters using Pearson correlation and multivariable regression models. (3) Results: Field-stained thin blood film microscopy and molecular identification revealed 8.98% and 10.18% infection rates, respectively. Phylogenetic analysis based on ITS-1 region sequences of the identified isolates showed close genetic associations with Indian isolates. The mean trypomastigote count observed in the infected buffaloes was 5.15 × 10⁶ (±5.3 × 10²)/µL of blood. The parasitemia loads were significantly correlated with the alterations in oxidative stress markers, DNA damage, and changes in hematobiochemical parameters. Infected animals exhibited significant (p < 0.05) alterations in oxidative stress biomarkers, including catalase, nitric oxide, and malondialdehyde concentrations. Noteworthily, a comet assay revealed a significantly (p < 0.0001) higher mean genetic damage index in the infected buffaloes (0.7 ± 0.04) compared with the healthy ones (0.196 ± 0.004). Alongside significant (p < 0.05) reductions in red cell indices, a marked elevation in leukocyte counts and serum hepatic enzyme levels was recorded in the affected buffaloes. (4) Conclusion: T. evansi isolates of buffaloes from Multan, Pakistan, have genetic similarities to Indian isolates. This study also revealed that higher parasitemia loads induce genotoxicity in the infected animals through oxidative stress and cause hematobiochemical alterations under natural field conditions.

Pb, a major environmental pollutant, can bio-accumulate in different tissues of food animals. The present study was accomplished to investigate the presence of Pb residues in various tissues of Lohi sheep (n = 360) and its deleterious effects. The blood and tissue samples (kidney, liver, and muscles) were collected from a local slaughterhouse for 6 weeks and analyzed through flame atomic absorption sepectrophotometry. The Pb concentration was determined to be maximum in kidney samples, followed by liver, serum, and muscle. In our analysis, 82.78% of muscle samples showed Pb concentrations below the maximum permissible limit (1 mg/kg) by the Australia New Zealand Food Authority (ANZFA). While values of serum ALT, AST, ALP, and urea were within normal range. In all the samples, RBC, TLC, PCV, and Hb values were significantly lower than control (P<0.05). Based on the above results, Lohi sheep seems to be quite resistant to the deleterious effects of Pb; however, its edible offal and lean muscle could pose a serious risk to public health.

Background/Objectives: Trypanosoma evansi (T. evansi) is an etiological agent of surra, and it causes significant economic losses in livestock. Rising trypanocide resistance demands alternatives that control parasitemia while mitigating oxidative and genotoxic damage. Therefore, the present study was designed to explore both the in vivo and in silico potential of Zingiber officinale (Z. officinale) as a novel phytotherapy to counter growing resistance against conventional trypanocides. Methods: Methanolic extract of Z. officinale (MZ) was orally administered at dosages of 200 mg/kg (MZ 200), 400 mg/kg (MZ 400), and 800 mg/kg (MZ 800) on a daily basis to the experimentally infected mice and compared against treated control (TC) and untreated control (UC) groups. After the infection, different parameters such as parasitemia counts, body weight changes, and the survival of infected mice were monitored for up to 7 days post-infection, while hematobiochemical parameters, oxidative stress profiles (catalase, malondialdehyde, and superoxide dismutase), and genotoxicity in brain tissues were compared at the end of the trial. Moreover, computational tools were used to predict the affinities of key bioactive compounds with twenty-one essential proteins of T. evansi. Results: The findings showed that the administration of MZ significantly (p < 0.05) reduced parasitemia and improved the survival rates in the experimentally infected mice in a dose-dependent manner. Noteworthy, significant (p < 0.05) improvements in hematological parameters and liver enzyme profiles were also recorded in MZ-treated groups. Compared to the untreated control, MZ-treated groups showed a significant amelioration in oxidative stress and genotoxicity in brain tissue in a dose-dependent fashion. The current study’s findings suggest that MZ potentially inhibits various essential proteins of T. evansi, including adenosine transporter-1, casein kinase, leucyl-tRNA synthetase, and multidrug resistance E protein. Among its constituents, 6-Isoshogaol and 6-Gingerol showed the most stable interactions in the molecular dynamics simulation. Conclusions: MZ efficiently reduced parasitemia, oxidative stress, and genotoxicity, and increased the survival rate in infected mice, suggesting it as a promising natural trypanicidal agent.

Bovine ephemeral fever (BEF) is a fatal viral disease predominantly affecting cattle and buffaloes. Infection results in a huge economic loss, especially due to the reduction in milk production. In Pakistan. There is a dearth of information on bovine ephemeral fever. The study was designed to detect and examine the effect of bovine ephemeral fever virus on hematological parameters among cattle in Swabi district, Khyber Pakhtunkhwa, Pakistan. A total of fifty blood samples were collected from suspected cattle and scrutinized for bovine ephemeral fever virus, using nested Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Hematological parameters were analyzed using hematology and chemistry analyzer. Of the 50 blood samples, 33 representing 66% were positive for bovine ephemeral fever virus through RT-PCR. A product size of 809 bp was observed in the reaction-I, while 505bp was obtained in reaction II. Neutrophils of infected cattle were significantly increased (10.44 +/- 1.87 x 10(9)/L) (p < 0.05), with a significant decrease (p < 0.05) in lymphocytes levels (2.90 +/- 0.97 x 10(9)/L). Changes in other blood parameters were non-significant. Furthermore, there was also a significant decrease (p < 0.05) in serum calcium level (7.84 +/- 0.16 mg/dL). We confirmed the detection of bovine ephemeral fever virus by nested reverse transcriptase polymerase chain reaction in Swabi, Khyber Pakhtunkhwa, Pakistan.