Cocktail of Theileria equi antigens for detecting infection in equines

El‒Sayed, Shimaa Abd El‒Salam; Rizk, Mohamed Abdo; Terkawi, Mohamad Alaa; Mousa, Ahmed Abdelmoniem; Said, El Said El Shirbini El; El-Sayed, Ghada M.; Fouda, Mohamed A.; Yokoyama, Naoki; Igarashi, Ikuo

doi:10.1016/j.apjtb.2015.09.001

Cited by 22 publications

(21 citation statements)

References 16 publications

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“…The Texas strain of B. bovis , [5, 6] the Argentina strain of B. bigemina , [6, 18] the German bovine strain of B. divergens [7, 8] and U.S. Department of Agriculture (USDA) strains of B. caballi [8] and T. equi [3, 6] were cultivated in purified bovine or equine red blood cells (RBCs) using a microaerophilic stationary-phase culture system [6, 18] and used for the in vitro experiment. The Munich strain of B. microti [4] was maintained by passage in the blood of BALB/c mice (CLEA, Tokyo, Japan) [19] and used in the in vivo study.…”

Section: Methodsmentioning

confidence: 99%

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Evaluation of the in vitro and in vivo inhibitory effect of thymoquinone on piroplasm parasites

et al. 2019

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BackgroundDeveloping new antibabesial drugs with a low toxic effect to the animal and with no resistance from Babesia parasites is in urgent demand. In this concern, the antimalarial, anticancer and antioxidant effect of thymoquinone (TQ), a phytochemical compound found in the plant Nigella sativa, has been reported. Therefore, in the present study, the antibabesial effect of this compound was evaluated on the growth of piroplasm parasites.ResultsSignificant inhibition (P < 0.05) of the in vitro growth of piroplasm parasites were observed after treatment by TQ with IC50 values of 35.41 ± 3.60, 7.35 ± 0.17, 0.28 ± 0.016, 74.05 ± 4.55 and 67.33 ± 0.94 μM for Babesia bovis, Babesia bigemina, Babesia divergens, Theileria equi and Babesia caballi, respectively. The in vitro inhibitory effect of TQ was significantly enhanced (P < 0.05) when used in combination with either diminazene aceturate on bovine Babesia and equine Babesia and Theileria cultures. In B. microti-infected mice, oral and intraperitoneal administrations of TQ showed significant (P < 0.05) inhibition of parasite growth at a dose of 70 mg/kg and 50 mg/kg, respectively, compared to the control group.ConclusionsThe obtained results indicate that thymoquinone might be a promising medicinal compound for use in the treatment of animal piroplasmosis.Electronic supplementary materialThe online version of this article (10.1186/s13071-019-3296-z) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

“…Moreover, Babesia divergens has a substantial impact on cattle health and productivity and has zoonotic importance [2]. In equines, the disease is caused by Theileria equi and B. caballi [3]. Generally speaking, babesiosis is typified in animals by high fever, hemolytic anemia and hemoglobinuria [1].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the in vitro and in vivo inhibitory effect of thymoquinone on piroplasm parasites

et al. 2019

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“…Myrrh oil was tested for its chemotherapeutic activity against B. bovis (Texas strain) [16], B. bigemina (Argentina strain) [23], B. caballi [26], and T. equi (United States Department of Agriculture) [4]. Parasites have been cultured in bovine or equine red blood cells using a continuous microaerophilic stationary phase culture system [16,23].…”

Section: In Vitro Growth Inhibition Assay and Viability Testmentioning

confidence: 99%

Myrrh Oil in Vitro Inhibitory Growth on Bovine and Equine Piroplasm Parasites and Babesia microti of Mice

et al. 2020

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The present experimental study was conducted for the assessment of the efficacy of in vitro inhibition of myrrh oil on the propagation of Babesia bovis, B. divergens, B. bigemina, Theileria equi, and B. caballi and in vivo efficacy on B. microti in mice through fluorescence assay based on SYBR green I. The culture of B. divergens B. bovis and was used to evaluate the in vitro possible interaction between myrrh oil and other commercial compound, such as pyronaridine tetraphosphate (PYR), diminazene aceturate (DA), or luteolin. Nested-polymerase chain reaction protocol using primers of the small-subunit rRNA of B. microti was employed to detect any remnants of DNA for studied parasitic species either in blood or tissues. Results elucidated that; Myrrh oil significantly inhibit the growth at 1% of parasitic blood level for all bovine and equine piroplasm under the study. Parasitic regrowth was inhibited subsequently by viability test at 2 µg/mL for B. bigemina and B. bovis, and there was a significant improvement in the in vitro growth inhibition by myrrh oil when combined with DA, PYR, and luteolin. At the same time; mice treated with a combination of myrrh oil/DA showed a higher inhibition in emitted fluorescence signals than the group that challenged with 25 mg/kg of diminazene aceturate at 10 and 12 days post-infection. In conclusion, this study has recommended the myrrh oil to treat animal piroplasmosis, especially in combination with low doses of DA.

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“…Moreover, Theileria equi ( T. equi ) and B. caballi are the causative agents of equine piroplasmosis, which is considered one of the most important protozoan diseases affecting horses, mules, and donkeys 1 . The disease is typified in general by high fever, anorexia, acceleration of the heart and respiratory rates, a marked drop in milk yield, general circulatory shock, hemoglobinuria, pale mucus membranes, and dry yellow feces or bloody stained feces; in severe cases, multi-organ failure might occur, resulting in death 2 , 3 . The drugs currently used in the field against babesiosis have been shown to have intolerable toxic effects, and resistance against them has been developed 4 .…”

Section: Introductionmentioning

confidence: 99%

Performance and consistency of a fluorescence-based high-throughput screening assay for use in Babesia drug screening in mice

Rizk

El‒Sayed

AbouLaila

et al. 2017

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In this study, we evaluated the validity of a fluorescence-based assay using SYBR Green I (SG I) stain for screening antibabesial compounds against B. microti in mice. Two different hematocrits (HCTs; 2.5% and 5%) were used. Correlating relative fluorescence units (RFUs) with parasitemia showed significant linear relationships with R2 values of 0.97 and 0.99 at HCTs of 2.5% and 5%, respectively. Meanwhile, the Z′ factors in a high-throughput screening (HTS) assay were within the permissible limit (≥0.5) at 2.5% HCT and lower than this value at 5% HCT. Taken together, the highest signal-to-noise (S/N) ratios were obtained at 2.5% HCT; therefore, we concluded that 2.5% was the best HCT for applying fluorescence assay in antibabesial drug screening in mice. Additionally, positive control mice and those treated with diminazene aceturate, pyronaridine tetraphosphate, and an allicin/diminazene aceturate combination showed peak parasitemia and fluorescence values on the same day post-inoculation. Moreover, using different concentrations of SG I revealed that the optimal concentration was 2x. In summary, considering that all experiments were applied under optimal laboratory conditions, fluorescence assay at 2.5% HCT using 2x SG I for B. microti parasite offers a novel approach for drug screening in mice.

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Cocktail of Theileria equi antigens for detecting infection in equines

Cited by 22 publications

References 16 publications

Evaluation of the in vitro and in vivo inhibitory effect of thymoquinone on piroplasm parasites

Evaluation of the in vitro and in vivo inhibitory effect of thymoquinone on piroplasm parasites

Myrrh Oil in Vitro Inhibitory Growth on Bovine and Equine Piroplasm Parasites and Babesia microti of Mice

Performance and consistency of a fluorescence-based high-throughput screening assay for use in Babesia drug screening in mice

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