Bioinformatics and expression analysis of the Xeroderma Pigmentosum complementation group C (XPC) of Trypanosoma evansi in Trypanosoma cruzi cells

Nucleotide excision repair (NER) acts repairing damages in DNA, such as lesions caused by cisplatin. Xeroderma Pigmentosum complementation group C (XPC) protein is involved in recognition of global genome DNA damages during NER (GG-NER) and it has been studied in different organisms due to its importance in other cellular processes. In this work, we studied NER proteins in Trypanosoma cruzi and Trypanosoma evansi, parasites of humans and animals respectively. We performed three-dimensional models of XPC proteins from T. cruzi and T. evansi and observed few structural differences between these proteins. In our tests, insertion of XPC gene from T. evansi (TevXPC) in T. cruzi resulted in slower cell growth under normal conditions. After cisplatin treatment, T. cruzi overexpressing its own XPC gene (TcXPC) was able to recover cell division rates faster than T. cruzi expressing TevXPC gene. Based on these tests, it is suggested that TevXPC (being an exogenous protein in T. cruzi) interferes negatively in cellular processes where TcXPC (the endogenous protein) is involved. This probably occurred due interaction of TevXPC with some endogenous molecules or proteins from T.cruzi but incapacity of interaction with others. This reinforces the importance of correctly XPC functioning within the cell.

Evaluation of an improved Giemsa staining technique for blood borne parasite in thick and thin blood film from dogs

Many a times the use of rapid diagnostic tests for blood borne parasites like trypanosomiasis and Babesiosis is increasingly being used, but the gold standard for its detection is still the use of microscopy both as reference and confirmatory diagnosis. To determine the effectiveness of the adjusted stock giemsa staining technique over the conventional methods. Venous Blood samples were collected from 10 dogs in EDTA and then used for the simultaneous preparation of two thin and thick smear slides, one stained according to Giemsa 1:20 dilution for 30 mins while the other was stained using the Stock Solution for 30seconds the diluted with buffered saline for 20seconds and rinsed. Fixation of the thin smear was done in a covered staining jar containing anhydrous methanol for 1 to 2 min, after which the slides were air-dried. From the result obtained from 10 dogs blood samples gotten from the veterinary clinic, the adjusted giemsa staining technique showed a positive differentiation when compared to the 1:20 dilution, a total of 7 blood samples tested positive for blood borne parasites, Trypanosoma evansi, Babesiosis cani and Heart worm. The highest percentage occurrence was T.evansi (40%), Babesiosis cani(20%) and Heart worm (10%).The adjusted Giemsa staining technique serves as a fast, easy and less complex alternative to the 1:20 dilution, where the solution has to be diluted from the stock solution and then stained, although the only disadvantage to this technique would be easy contamination of the stock solution, but the advantages here is that it saves time, quicker result output and better differentiation microscopically.

Single-cell transcriptome profiling and the use of AID deficient mice reveal that B cell activation combined with antibody class switch recombination and somatic hypermutation do not benefit the control of experimental trypanosomosis

Salivarian trypanosomes are extracellular protozoan parasites causing infections in a wide range of mammalian hosts, with Trypanosoma evansi having the widest geographic distribution, reaching territories far outside Africa and occasionally even Europe. Besides causing the animal diseases, T. evansi can cause atypical Human Trypanosomosis. The success of this parasite is attributed to its capacity to evade and disable the mammalian defense response. To unravel the latter, we applied here for the first time a scRNA-seq analysis on splenocytes from trypanosome infected mice, at two time points during infection, i.e. just after control of the first parasitemia peak (day 14) and a late chronic time point during infection (day 42). This analysis was combined with flow cytometry and ELISA, revealing that T. evansi induces prompt activation of splenic IgM+CD1d+ Marginal Zone and IgMIntIgD+ Follicular B cells, coinciding with an increase in plasma IgG2c Ab levels. Despite the absence of follicles, a rapid accumulation of Aicda+ GC-like B cells followed first parasitemia peak clearance, accompanied by the occurrence of Xbp1+ expressing CD138+ plasma B cells and Tbx21+ atypical CD11c+ memory B cells. Ablation of immature CD93+ bone marrow and Vpreb3+Ly6d+Ighm+ expressing transitional spleen B cells prevented mature peripheral B cell replenishment. Interestingly, AID-/- mice that lack the capacity to mount anti-parasite IgG responses, exhibited a superior defense level against T. evansi infections. Here, elevated natural IgMs were able to exert in vivo and in vitro trypanocidal activity. Hence, we conclude that in immune competent mice, trypanosomosis associated B cell activation and switched IgG production is rapidly induced by T. evansi, facilitating an escape from the detrimental natural IgM killing activity, and resulting in increased host susceptibility. This unique role of IgM and its anti-trypanosome activity are discussed in the context of the dilemma this causes for the future development of anti-trypanosome vaccines.

Mortalidad de equinos por Trypanosoma evansi en Argentina. Diagnóstico parasitológico y molecular

Trypanosoma evansi, el agente causal de la tripanosomosis, afecta a una amplia variedad de animales domésticos y silvestres, siendo los equinos los principales enfermados. El objetivo de este trabajo fue describir un brote de tripanosomosis por T. evansi en veinte (20) equinos con sintomatología caracterizada por una progresiva pérdida de peso, inapetencia, hipertermia intermitente, debilidad, linfoadenomegalia, mucosas pálidas, secreciones nasales, epífora, edemas en zona de pecho, abdomen y miembros posteriores. El diagnóstico parasitológico y molecular fue positivo para T. evansi. Los animales fueron tratados con aceturato de diminacina sin resultados satisfactorios. Este trabajo es el primer reporte de mortandad de equinos por T. evansi con diagnóstico parasitológico y molecular, en la localidad de Monte Caseros, Corrientes, Argentina.

HISTOLOGICAL CHANGES OF ADRENAL GLAND IN RABBITS DURING THE EXPERIMENTAL INFECTION WITH TRYPANOSOMA EVANSI AND AFTER TREATMENT WITH NAGANOL

The structural – functional changes in the adrenal glands associated with experimental infection with Trypanosoma evansi and the reversed changes after the treatment of rabbits with anti-trypanosomal drugs (Naganol, Bayer, 205) were studied. Thirty adult New Zealand male rabbits were divided into two groups: control group which include 12 rabbits while the infected group include 13 rabbits. Rabbits of infected group were injected I. V with 10 trypanosoma on 0.2 ml mice blood diluted with AL -sever's solution while rabbits of control group were injected with trypansoma free diluted mice blood. Forty-two days after infection the infected group were treated with neganol 10% sol. (20mg/ kg B.W, i.v) than all the rabbits were killed after 42 days of treatment. Microscopic examination of the hematoxylin-eosin stained adrenal gland revealed occurrence of degenerative changes of the zona fosciculata-reticulaties infltration of mononuclear cells, red blood cells and focal coagulative necrosis in both zone when compared with those of the control group, while treated group showed focal regeneration of secretort cells of both zone. It is concluded there were revers of structural-functional changes after treatment to the normal state.

Sero-prevalence and immunological characterization of Trypanosoma evansi infection in livestock of four agro-climatic Zones of Himachal Pradesh, India

Trypanosoma evansi, a hemoflagellate protozoan parasite causes wasting disease called surra in wide range of animals. Although, the organism has been reported from various parts of the India, data generated from organized epidemiological study is still in infancy in majority states of India. In the present study, livestock of Himachal Pradesh, India was targeted for epidemiological investigation of T. evansi infections. A total of 440 equines and 444 cattle serum samples were collected from four agro-climatic Zones. Further, serum samples of 280 buffaloes from three different agro-climatic Zones of Himachal Pradesh were also collected and evaluated for presence of T. evansi infection by indirect ELISA. Data generated showed higher prevalence in buffalo (23.57%) followed by cattle (22.52%) and equines (1.82%). Disease was found to be more prevalent (P < 0.05) in cattle of lower altitude as compared to those of higher altitudes. No significant variation was seen in prevalence of disease on the basis of age and sex of the animals. Serum biochemical analysis revealed increased levels of BUN in T. evansi infected equines. Levels of liver function enzymes such as ALT/GGT and AST were found to be significantly elevated (P < 0.01) in infected animals whereas glucose levels were significantly lower in surra infected animals as compared to non-infected animals. Western blot analysis of whole cell lysate (WCL) antigen of T. evansi using surra infected serum samples of equines showed immunodominant bands in the range of 100-25 kDa. Surra infected bovine serum samples recognized polypeptide bands in the range of 85-32 kDa, including protein clusters of 52-55 and 48-46 kDa. Poypeptide cluster of 62-66 kDa was found common to serum samples of bovines and equines from all agro-climatic Zones. Animal trypanosomosis was found to be highly prevalent in livestock of Himachal Pradesh and thus there is dire need for designing of proper control strategies against surra.