Qais A. H. Majeed

The present study aimed to evaluate the in vitro, in vivo, and safety of Stachys lavandulifolia Vahl. methanolic extract (SLME) against acute toxoplasmosis caused by Toxoplasma gondii RH strain in mice. Methods: MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to evaluate the in vitro effect of the SLME on T. gondii tachyzoites. Totally, 72 male BALB/c mice (40 mice for in vivo evaluation of SLME and 32 mice for its toxicity effects on liver and kidney serum enzymes) were used for the present investigation. At first, 40 mice were orally pre-treated with the SLME at doses of 25, 50, and 75 mg/kg/day for two weeks. Mice were checked daily, and the rate of survival and the mean number of tachyzoites were recorded. Liver lipid peroxidation (LPO) and nitric oxide (NO) levels, the effects on kidney and liver function, as well as the expression level of the proinflammatory cytokines such as interleukin-1β (IL-1β) and interferon-γ (IFN-γ), were studied by the quantitative real-time PCR. Flow cytometry analysis was performed on the effects of SLME on the detection of apoptotic and necrotic cells in T. gondii tachyzoites. Results: The SLME at the concentrations 75 and 150 µg/mL completely killed the tachyzoites after 2 hr of incubation. SLME at 25, 50, and 75 mg/kg/day increased the survival rate of infected mice by the sixth, seventh, and eighth days, respectively. SLME also significantly (p < 0.05) decreased the LPO and NO levels and upregulated the IL-1β and IFN-γ mRNA gene expression levels, whereas no considerable change was observed in the serum level of kidney and liver enzymes. Flow cytometry analysis revealed the prompted early and late apoptosis after exposure to T. gondii tachyzoites with various concentrations of SLME. Conclusion: We found the relevant in vitro anti-Toxoplasma effects of SLME against T. gondii. Moreover, the results confirmed the promising in vivo prophylactic effects of SLME. SLME provokes the innate immune system, induces apoptosis, modulates the proinflammatory cytokines, and inhibits hepatic injury in infected mice. With all these descriptions, further surveys are required to support these findings and elucidate this plant’s possible mechanisms of action.

Cryptosporidium is a worldwide enteric protozoan parasite that causes gastrointestinal infection in animals, including humans. The most notable species is Cryptosporidium parvum because of its zoonotic importance; it is also the leading cause of cryptosporidiosis in preweaned calves. A cross-sectional study was conducted to determine the prevalence of Cryptosporidium infection, investigate the potential risk factors, and use molecular diagnosis to identify the predominant Cryptosporidium spp. in preweaned calves in Kuwait. Of 175 preweaned calves, Cryptosporidium antigens were detected in 58 (33.1%) using rapid lateral immunochromatography assay (IC). Calves less than one month of age (OR = 4.32, p = 0.0001) and poor hygiene (OR = 2.85, p = 0.0075) were identified as significant risk factors associated with Cryptosporidium infection. Molecular identification revealed that C. parvum (62.8%) was the dominant species infecting preweaned calves in Kuwait. In contrast, C. bovis and C. andersoni were recorded at 5.7% and 2.9%, respectively. All C. parvum gp60 nucleotide sequences were subtype IIaA15G2R1. Calves could be a source of C. parvum infection due to the similarity of the subtypes recorded previously in Kuwaiti children and preweaned calves in this study. Therefore, more research is needed to understand the Cryptosporidium transmission cycle in Kuwait.