Fighting eimeriosis by using the anti-eimerial and anti-apoptotic properties of rhatany root extract.

Background: Over the last decade, extensive use of coccidiostats to treat and control Eimeria infection has developed drug resistance, prompting the search for new alternative therapies. Rhatany is proven to have various pharmacological properties. Objective: The present study aimed to in vitro and in vivo evaluate the effect of Rhatany roots extract (RRE) as an anti-eimerial and anti-apoptotic agent against murine eimeriosis induced by Eimeria papillata. Methods: Phytochemical screening by gas chromatography-mass spectrometry analysis (GC-MS) was used to detect active compounds in RRE. In vitro anti-eimerial activity of RRE (200, 100, 50 mg/ml), amprolium, phenol, Dettol™, and formalin were studied after incubation with non-sporulated Eimeria oocysts. For the in vivo study, twenty-five male C57BL/6 mice were randomly allocated into five groups. Animals in the first group were just given distilled H2O, while those in the second group were given 200 mg/kg RRE for 5 days. The Eimeria parasite's oocysts were infected into the third, fourth, and fifth groups. For treatment, RRE (200 mg/kg) and amprolium (120 mg/kg) were orally given to the 4th and 5th groups for five days, respectively. All mice were euthanized, on day 5 post-infection, to collect the jejunal tissues under study. Investigations were undertaken into the oocyst output in feces and goblet cells in mice jejuna. Assays for glutathione peroxidase (GPx), hydrogen peroxide (H2O2), and myeloperoxidase (MPO) were also performed. In jejunal tissue, cysteine aspartic acid protease-3 (Caspase-3) was counted using immunohistochemistry, while BCL2-associated X protein (Bax) and B-cell lymphoma-2 (BCL-2) were assayed using ELISA. In addition, mRNA expression of the goblet cell response gene (MUC2) was detected using real-time PCR. Results: Phytochemical screening by GC-MS demonstrated the presence of 22 compounds in the RRE. The in vitro study revealed that RRE significantly inhabited the oocyst sporulation in a dose-dependent manner. By day 5 after infection with the Eimeria parasite, the number of oocysts in mice feces was significantly reduced after RRE treatment (1.308 × 106 ± 1.36 × 105 oocysts/g feces) compared to the infected group (5.387 × 106 ± 4.29 × 105 oocysts/g feces). Moreover, the Eimeria infection reduced the number of goblet cells of mice jejuna and its specific gene, MUC2. The treatment with RRE increased the number of goblet cells/villus from 3.45 ± 0.17 to 6.04 ± 0.23, associated with upregulation for MUC2 from 0.26 to 2.39-fold. Also, the Eimeria experimental infection lowered the activity of the antioxidant enzyme represented by GPx (23.99 ± 3.68 mg/g tissue), while increasing the stress parameters of hydrogen peroxide (0.07 ± 0.01 mM/g) as well as the activity of MPO (66.30 ± 3.74 U/mg). The production of apoptotic markers including Caspase-3 (68.89 ± 2.67 U/g) and Bax (159.05 ± 6.50 pg/ml) was significantly elevated while decreasing the anti-apoptotic marker of BCL2 (0.42 ± 0.07 pg/ml). Our study proved that RRE significantly reduced oxidative stress, and apoptotic markers as well as the inflammatory activity of MPO. Also, antioxidant enzyme and anti-apoptotic activity in the jejunum of E. papillata-infected mice were enhanced after RRE treatment. Conclusion: Our study highlights the potential of RRE as a natural solution for coccidiosis management by modulating apoptosis in E. papillata host cells. However, further research is needed to fully understand the underlying mechanisms and enhance our understanding of its therapeutic efficacy.

Krameria lappacea root extract's anticoccidial properties and coordinated control of CD4 T cells for IL-10 production and antioxidant monitoring.

Introduction: Recently, the use of botanicals as an alternative to coccidiostats has been an appealing approach for controlling coccidiosis. Therefore, this study was conducted to evaluate the potential role of aqueous methanolic extract (200 mg/kg) of Krameria lappacea (roots) (KLRE) against infection induced by Eimeria papillata. Methods: A total of 25 male C57BL/6 mice were divided into five groups (I, II, III, IV, and V). On 1st day of the experiment, all groups except groups I (control) and II (non-infected-treated group with KLRE), were inoculated orally with 103 sporulated E. papillata oocysts. On the day of infection, group IV was treated with KLRE. Group V served as an infected-treated group and was treated with amprolium (coccidiostat). Results: Treatment with extract and coccidiostat was continued for five consecutive days. While not reaching the efficacy level of the reference drug (amprolium), KLRE exhibited notable anticoccidial activity as assessed by key criteria, including oocyst suppression rate, total parasitic stages, and maintenance of nutrient homeostasis. The presence of phenolic and flavonoid compounds in KLRE is thought to be responsible for its positive effects. The Eimeria infection increased the oxidative damage in the jejunum. KLRE treatment significantly increased the activity of catalase and superoxide dismutase. On the contrary, KLRE decreased the level of malondialdehyde and nitric oxide. Moreover, KLRE treatment decreased macrophage infiltration in the mice jejunal tissue, as well as the extent of CD4 T cells and NFkB. E. papillata caused a state of systemic inflammatory response as revealed by the upregulation of inducible nitric oxide synthase (iNOs)-mRNA. Upon treatment with KLRE, the activity of iNOs was reduced from 3.63 to 1.46 fold. Moreover, KLRE was able to downregulate the expression of pro-inflammatory cytokines interferon-γ, nuclear factor kappa B, and interleukin-10 -mRNA by 1.63, 1.64, and 1.38 fold, respectively. Moreover, KLRE showed a significant reduction in the expression of IL-10 protein level from 104.27 ± 8.41 pg/ml to 62.18 ± 3.63 pg/ml. Conclusion: Collectively, K. lappacea is a promising herbal medicine that could ameliorate the oxidative stress and inflammation of jejunum, induced by E. papillata infection in mice.

Anticoccidial and jejunum-protective effects of Krameria lappacea roots extract on experimental Eimeria papillata infection.

Coccidiosis is a protozoan parasitic disease caused by Eimeria species and affects wild and domestic animals. Coccidiostats are currently available to control this disease, although drug resistance has been confirmed for all of them. As a result, there is an urgent need to identify eco-friendly agents to control and treat this disease. This study aimed to investigate the ameliorative role of the Krameria lappacea roots extract (KLRE) on the outcome of coccidiosis induced by Eimeria papillata. Male C57BL/6 mice were divided into seven groups (5 mice/group), as follows: Group 1: noninfected-nontreated (control group), Group 2: noninfected-treated group with KLRE (200 mg/kg), Group 3: infected-nontreated group, Group 4: infected-treated group with KLRE (50 mg/kg), Group 5: infected-treated group with KLRE (100 mg/kg), Group 6: infected-treated group with KLRE (200 mg/kg), and Group 7: infected-treated group with amprolium (120 mg/kg). Groups (3-7) were inoculated orally with 1 × 103 sporulated E. papillata oocysts. One hour after infection, groups (4-6) were daily treated for 5 days with KLRE and amprolium. On day 5 postinfection, oocyst output was determined, and mice were euthanized for the collection of jejuna then preparation of histological sections and jejunal homogenate was used for the determination of biochemical and oxidative damage markers. The coccidial infection induced weight loss of mice by 3.971%, which improved after KLRE to -1.512%. After KLRE treatment, the rate of feed intake was improved to be 52.21 ± 2.30 than those in infected group (40.47 ± 2.25). Oocyst output was significantly reduced in mice treated with KLRE (1.308 × 106 oocysts/g.feces) compared with those in the infected group (5.387 × 106 oocysts/g.feces). E. papillata infection induced marked histological alterations within jejunum tissue. After treatment, KLRE was able to impair the development of parasite stages (meronts, gamonts, and developing oocysts) in the jejunum through a significant reduction of number and size in comparison with the infected group. Infection with E. papillata induced a disturbance in the nutrient absorption in the jejunal mice tissue, which improved after the treatment with KLRE and amprolium. Also, KLRE counteracted significantly the E. papillata-induced loss of reduced glutathione and total antioxidant capacity. Our findings indicate that KLRE could be used as an alternative to the available coccidiostats currently available. RESEARCH HIGHLIGHTS: Krameria lappacea exhibit significant anticoccidial and antioxidant activities induced by E. papillata infection. Krameria lappacea exhibit significant improvement in the pathological alterations of the jejunal tissue induced by E. papillata infection.