Comparative study of three novel ion exchange resins with DEAE-cellulose for the purification of Trypanosoma evansi.

Ion exchange chromatography is a method that uses the different surface charges of trypanosomes and blood cells to separate them. This makes it possible to use molecular and immunological methods to diagnose or study these protozoans. DEAE-cellulose resin is commonly used to perform this method. The goal of this study was to compare three novel chromatographic resins designated as PURIFICA™ (Y-C2N®, Y-HONOH®, and Y-CNC3®). The resins were evaluated based on their ability to isolate the parasite, purification time, examination of parasite viability and morphology, and trypanosome recovery potential after passing through the columns. In terms of the evaluated parameters, there was no significant difference between DEAE-cellulose and the three tested resins in most experiments. However, PURIFICA™ (Y-C2N®, Y-HONOH®, and Y-CNC3®) resins are less expensive and easier to prepare than DEAE-Cellulose, making them an alternative for the purification of Trypanosoma evansi.

First record of Trypanosoma evansi DNA in Dichelacera alcicornis and Dichelacera januarii (Diptera: Tabanidae) flies in South America.

BACKGROUND: Trypanosoma evansi infects a large number of wild and domestic animals and causes a spoliative disease known as surra. It is mechanically transmitted, mainly by biting flies of the genera Tabanus and Stomoxys. The detection of T. evansi DNA in the feeding apparatus of Dichelacera alcicornis and Dichelacera januarii from South America is reported, to the best of our knowledge, for the first time. METHODS: Tabanids were collected weekly from February 2018 to February 2019 from two sites. The feeding apparatus was removed and DNA extraction, polymerase chain reaction and sequencing were performed. RESULTS: A 205-base pair fragment of the variant surface protein RoTat 1.2 gene, confirmed by DNA sequencing, was amplified from the feeding apparatus of D. alcicornis and D. januarii. CONCLUSIONS: This is, to the best of our knowledge, the first record of T. evansi DNA in South American tabanids.

Trypanosoma evansi secretome carries potential biomarkers for Surra diagnosis.

Trypanosoma evansi is a parasite that is phylogenetically close to Trypanosoma brucei and is the causative agent of a disease known as surra. Surra is responsible for a high mortality rate in livestock and large economic losses in the Americas, Africa, and Asia. This work aimed to analyze in vitro secreted proteins from T. evansi and identify potential treatment and diagnostic biomarkers for surra diagnosis. Two groups were used. In one group the parasites were purified using a DEAE-Cellulose column and maintained in a secretion medium while in the other group the parasites were not purified. Each group was further divided to be maintained at either 37 °C or 27 °C. We identified 246 proteins through mass spectrometry and found that the temperature appears to modulate protein secretion. We found minimal variations in the protein pools from pure and non-purified sets. We observed an emphasis on proteins associated to vesicles, glycolysis, and cellular homeostasis through the enrichment of GO. Also, we found that most secretome proteins share homologous proteins with T. b. brucei, T. b. gambiense, T. vivax, T. equiperdum, and T. b. rhodesiense secretome but unique T. evansi epitopes with potential biomarkers for surra diagnosis were detected. SIGNIFICANCE: Trypanosoma evansi is a parasite of African origin that is phylogenetically close to Trypanosoma brucei. As with other trypanosomatids and blood parasites, its infection causes non-pathognomonic symptoms, which makes its diagnosis difficult. One great problem is the fact that no diagnostic test differentiates between Trypanosoma equiperdum and T. evansi, which is a problem in South America and Asia, and Africa. Thus, it is urgent to study the biochemistry of the parasite to discover proteins that can be used for differential diagnosis or be possible therapeutic targets. In addition, the study of the secretome can point out proteins that are used by the parasite in its interactions with the host, helping to understand the progression of the disease.

Neospora caninum-associated abortions in cattle from Southern Brazil: Anatomopathological and molecular characterization.

This study aimed to evaluate the N. caninum associated abortions in cattle in the state of Santa Catarina, in the southern Brazil. Aborted bovine fetuses were necropsied, submitting organ samples for histopathological evaluation. Brain fragments were analyzed by polymerase chain reaction (PCR). The diagnosis of abortion due to N. caninum was established through histopathology and molecular analysis in 53.84% (28/52) of the cases, with PCR detection in 71.42% (20/28). The histopathological evaluation showed lesions in 75% of the cases, characterized by mononuclear necrotizing encephalitis, mononuclear myocarditis, mononuclear myositis, mixed placentitis, and mononuclear pneumonia. Neospora caninum was the primary etiological agent associated with causes of abortion in cattle in the present study.

Trimming and washing poultry carcass to reduce microbial contamination: a comparative study.

The objective of this study was to compare the efficiency of washing and trimming broiler carcasses to reduce bacterial contamination. At the postevisceration site, 100 broiler carcasses were collected during 4 visits to a slaughterhouse in Santa Catarina State, Brazil. Birds were from the same flock, age, and approximately 2.4 kg of weight. Groups were as follows: group 1, with fecal contamination; group 2, without fecal contamination; group 3, with fecal contamination and trimmed; group 4, with fecal contamination and washed; group 5, with fecal contamination, and washed and trimmed. Carcass washings were performed with at least 1.5 L/bird of potable water (0.5 to 1 mg/kg of residual chlorine) at room temperature (20-25°C) using spray cabinets with 44 spray nozzles distributed into 2 chambers (pressure of 2 kgf/cm(2) and 4 kgf/cm(2)). Washed carcasses (trimmed or not) showed significantly (P < 0.05) lower counts of aerobic mesophiles (plate count agar) on the third evaluation, and even lower (P < 0.01) counts for total coliforms (CT) and fecal coliforms (Escherichia coli). Trimmed carcasses showed significantly lower counts (P < 0.05) for plate count agar; however, we observed higher counts for E. coli (P < 0.05). The association of both treatments (washing and trimming) showed significantly higher (P < 0.05) counts for coliforms (CT and E. coli). We can conclude that the washing method is overall more efficient than the trimming method to decontaminate chicken carcasses at the postevisceration site. Hopefully, our findings can help poultry companies to minimize production costs by applying the washing method for carcass decontamination.