Molecular identification of new Trypanosoma evansi type non-A/B isolates from buffaloes and cattle in Indonesia.

Trypanosoma evansi is reportedly divided into two genotypes: types A and B. The type B is uncommon and reportedly limited to Africa: Kenya Sudan, and Ethiopia. In contrast, type A has been widely reported in Africa, South America, and Asia. However, Trypanosoma evansi type non-A/B has never been reported. Therefore, this study aims to determine the species and genotype of the Trypanozoon subgenus using a robust identification algorithm. Forty-three trypanosoma isolates from Indonesia were identified as Trypanosoma evansi using a molecular identification algorithm. Further identification showed that 39 isolates were type A and 4 isolates were possibly non-A/B types. The PML, AMN-SB1, and STENT3 isolates were likely non-A/B type Trypanosoma evansi isolated from buffalo, while the PDE isolates were isolated from cattle. Cladistic analysis revealed that Indonesian Trypanosoma evansi was divided into seven clusters based on the gRNA-kDNA minicircle gene. Clusters 6 and 7 are each divided into two sub-clusters. The areas with the highest genetic diversity are the provinces of Banten, Central Java (included Yogyakarta), and East Nusa Tenggara. The Central Java (including Yogyakarta) and East Nusa Tenggara provinces, each have four sub-clusters, while Banten has three.

Ovicidal activity and cytotoxicity of ethanolic extract of turmeric (Curcuma longa) and green tea (Camellia sinensis) to treat digestive parasite of sheep.

Background: The Trichuris eggs are collected from naturally infected sheep. Natural antihelmintics such as herbal medicines are needed as an alternative, such as natural compounds from endemic plants. Aim: This present study aims to evaluate the ovicidal activity and cytotoxicity effects of ethanolic extract of Curcuma longa (EECL) and Camelia sinensis (EECS) as a biological anthelmintic against the egg of Trichuris sp. Methods: The Trichuris eggs are collected from naturally infected sheep. CMC-Na solution 1% was used as a control. The treatments were 0.12% EECL; 0.24% EECL; 0.15% EECS; 0.30% EECS; a combination of 0.12% EECL and 0.30% EECS; a combination of 0.24% EECL; and 0.15% EECS. Ovicidal activity testing by microscopic examination of eggs treated using different concentrations of EECL extract, EECS, and a combination of them. They were exposed for various times (7, 14, 21, and 28 days) and incubated at room temperature. Results: The study shows that a combination of C. longa extract and tea extract exhibits good ovicidal anthelmintic activity against Trichuris sp. in sheep. Cytotoxicity examination using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test. Based on MTT data processed using regression analysis, the obtained LC50 from the administration of EECL, EECS, and a combination of both in a ratio of 1:1, 2:2, 1:2, and 2:1. The combination of EECL extract and EECS with the highest concentration produced cell viability of 28.46%, 17.25%, 56.01%, and 46.47%, respectively. Conclusion: It can be concluded that the most cytotoxic ingredient is found in the combination of EECL and EECS (2:2) at 17.25% and the safest is in the ratio (1:2) at 56.01%.

Tissue cysts and serological detection toxoplasmosis among wild rats from Surabaya, East Java, Indonesia.

Background: The protozoan Toxoplasma gondii is the source of zoonosis toxoplasmosis and causes public health problems throughout the world. Environmental contamination by oocysts excreted by cats as definitive hosts affects the spread of this disease. Wild rats as rodents can be used as an indicator of environmental contamination by oocysts, considering that rats have a habit of living in dirty environments and can be infected by oocysts from the environment. Aim: This study aims to detect toxoplasmosis from tissue cysts and serological tests in wild rats as an indicator of environmental contamination in Surabaya. Methods: A total of 100 wild rats collected from Surabaya were collected in five areas (West, East, Central, North, and South of Surabaya) obtained from three trapping locations: housing, dense settlements, and markets. All samples were examined microscopically for parasitological tests through the brain tissue samples, and the serum was examined using the toxoplasma modified agglutination test to detect the presence of IgG and Immunoglobulin M (IgM). Results: This research used 100 wild rat samples, 77 Rattus tanezumi and 33 Rattus norvegicus, with evidence of 31% in serology and active infection with 19% tissue cyst. The results showed that the seroprevalence of T. gondii in wild rats was 31% (30% for IgG and 1% for IgM). Tissue cysts in the rat brain samples tested were 19% (19/100). The IgG prevalence rate in female rats was 25% (8/32), while for males, it was 32.3% (22/68). The highest seropositive IgG from densely populated settlements was 50%, markets were 25.8%, and housing was 12.1%. The highest seropositive IgM from densely populated settlements was 2.8%. Population density and the presence of cats are factors supporting the high seropositive rate at the trapping location. Conclusion: This study revealed that there has been toxoplasmosis contamination in Surabaya with evidence of 31% in serology and active infection with 19% tissue cyst. It is necessary for controlling with surveillance in cats to prevent transmission in humans.

Molecular identification of new Trypanosoma evansi type non-A/B isolates from buffaloes and cattle in Indonesia / Identificação molecular de novos isolados de Trypanosoma evansi tipo não A/B em búfalos e gado na Indonésia

Abstract Trypanosoma evansi is reportedly divided into two genotypes: types A and B. The type B is uncommon and reportedly limited to Africa: Kenya Sudan, and Ethiopia. In contrast, type A has been widely reported in Africa, South America, and Asia. However, Trypanosoma evansi type non-A/B has never been reported. Therefore, this study aims to determine the species and genotype of the Trypanozoon subgenus using a robust identification algorithm. Forty-three trypanosoma isolates from Indonesia were identified as Trypanosoma evansi using a molecular identification algorithm. Further identification showed that 39 isolates were type A and 4 isolates were possibly non-A/B types. The PML, AMN-SB1, and STENT3 isolates were likely non-A/B type Trypanosoma evansi isolated from buffalo, while the PDE isolates were isolated from cattle. Cladistic analysis revealed that Indonesian Trypanosoma evansi was divided into seven clusters based on the gRNA-kDNA minicircle gene. Clusters 6 and 7 are each divided into two sub-clusters. The areas with the highest genetic diversity are the provinces of Banten, Central Java (included Yogyakarta), and East Nusa Tenggara. The Central Java (including Yogyakarta) and East Nusa Tenggara provinces, each have four sub-clusters, while Banten has three.

Resumo Trypanosoma evansi é reportado como dividido em dois genótipos: tipos A e B. O tipo B é incomum e reportado como limitado à África: Quênia, Sudão e Etiópia. Em contraste, o tipo A tem sido amplamente relatado na África, América do Sul e Ásia. No entanto, Trypanosoma evansi tipo não-A/B nunca foi relatado. Portanto, este estudo tem como objetivo determinar a espécie e o genótipo do subgênero Trypanozoon, utilizando-se um algoritmo robusto de identificação. Quarenta e três isolados de tripanosoma da Indonésia foram identificados como Trypanosoma evansi, usando-se um algoritmo de identificação molecular. A identificação adicional mostrou que 39 isolados eram do tipo A e 4 isolados eram, possivelmente, do tipo não A/B. Os isolados PML, AMN-SB1 e STENT3 foram, provavelmente, Trypanosoma evansi do tipo não A/B isolado de búfalos, enquanto os isolados de PDE foram isolados de bovinos. A análise cladística revelou que o Trypanosoma evansi indonésio foi dividido em sete grupos baseados no gene do minicírculo gRNA-kDNA. Os clusters 6 e 7 foram divididos cada um em dois subclusters. As áreas com maior diversidade genética são as províncias de Banten, Java Central (incluindo Yogyakarta) e East Nusa Tenggara. As de Java Central (incluindo Yogyakarta) e East Nusa Tenggara têm, cada uma, quatro subgrupos, enquanto Banten tem três.