Toxocariasis is a neglected zoonosis that affects children and adults. Recombinant proteins have been widely investigated for diagnosis, achieving high sensitivity and specificity in an overall population; however, little is known about age as a factor in its application. This study aims to investigate the diagnostic potential of Toxocara canis TES-30 and TES-120 recombinant proteins in humans, differentiating between its performance in children and adults. Serum samples collected from children and adults seropositive to Toxocara spp. were tested with indirect ELISA using T. canis TES-30 and TES-120 recombinant proteins produced in Escherichia coli. While rTES-30 sensitivity was not affected by age (81.8% in children and 87% in adults), rTES-120 sensitivity severely decreased in children to only 63.6%, down from 95.7% in adults. Furthermore, the sensitivity of rTES-30 increased to 97.8% after Western blotting confirmation. High specificity (>94%) against other geohelminths was reported for both recombinant proteins. Our study favors the use of rTES-30 with total IgG as the primary antibody in an indirect ELISA assay as a tool for epidemiological human studies.

Recombinant proteins are a suggested alternative for the diagnosis of toxocariasis. The current Escherichia coli recombinant protein overexpression system usually produces insoluble products. As an alternative, yeast such as Pichia pastoris have secretory mechanisms, which could diminish the cost and time for production. This study aimed to produce recombinant proteins in Pichia pastoris and verify their sensibility and specificity in an indirect ELISA assay. Two sequences (rTES-30 and rTES-120) of Toxocara canis excretory-secretory antigens were cloned in a pPICZαB vector and expressed in P. pastoris KM71H. Sera samples collected from human adults infected by Toxocara spp. were tested by indirect ELISA using rTES-30 and rTES-120 as antigens. Recombinant proteins were detected at 72 hours after induction, in the HIGHLIGHTS  Pichia pastoris rTES evaluated as an alternative tool for toxocariasis diagnosis.  No difference between sensitivities of glycosylated and partial glycosylated rTES.  High-throughput expression and in vivo purification of rTES in Pichia pastoris. 2 Santos, L.M.; et al.

The objective of this study was to obtain information with scientific validity regarding the probable physiological alterations that sheep may suffer in the period prior to slaughter and its relationship with welfare conditions and maintenance of homeostasis. Twenty-nine castrated male lambs were used, clinically healthy, with no defined breed, and aged up to one year. Before slaughter, the heart and respiratory frequencies were measured, as well as the rectal temperature. At the time of bleeding, a blood sample was collected for analysis of hematological parameters (lactic acid, glucose, hematocrit). The mean value found for the rectal temperature was 39.30 ± 0.45 °C, for respiratory rate of 64.10 ± 28.52mov/min, heart rate of 106.90 ± 30.57bat/min, lactic acid of 4.81 ± 1.04mmol/L, glucose of 69.79 ± 10.33mg/dL and hematocrit of 38.93 ± 3.28%. The database analysis showed a low and positive correlation between respiratory rate and rectal temperature. It was concluded that lambs kept at high temperatures showed an increase in respiratory and cardiac frequencies, as well as an intensification of the stress condition reflected by the increase in plasma lactic acid levels.