The Mating Type (MAT) and Virulence of Sporothrix schenckii sensu stricto Isolates Maintained in Culture Collection.

In an attempt to determine the mating type of different Sporothrix schenckii sensu stricto isolates that remained viable after a long period of preservation in a culture collection and to correlate them with the degree of virulence/pathogenicity, a PCR technique using primers designed for the sequences of MAT1-1-1 and MAT1-2-1 genes and a murine experimental model were used. The results showed that there was no correlation between the mating type and virulence among the isolates. Furthermore, different degrees of virulence/pathogenicity, ranging from high to low, were found among them based on different virulence parameters. It was assumed that the long period of preservation favored the changes, yielding the isolation of variants. Thus, we believe that new technologies for studies on factors can improve our knowledge of the pathogenesis of sporotrichosis.

Applicability of the Monocyte Activation Test (MAT) for hyperimmune sera in the routine of the quality control laboratory: Comparison with the Rabbit Pyrogen Test (RPT).

Pyrogen tests are safety assays performed during the routine quality control of injectable products required by regulatory agencies. Currently, there are three available testing possibilities: 1) the Rabbit Pyrogen Test (RPT); 2) the Bacterial Endotoxin Test (BET); and 3) test systems using human whole-blood or monocytes, termed Monocyte Activation Test (MAT). Although BET is often considered as a replacement for the animal test, it is unable to detect pyrogens other than endotoxin. MAT is based on the human fever reaction and thus, most closely reflects the human response. The aim of this study was to conduct a parallel comparison of the RPT and MAT for hyperimmune sera (HS) batches analyzed during the routine of a quality control laboratory. MAT was performed in the same 43 batches of HS previously tested using RPT. The results showed that MAT presented 100% sensitivity and approximately 85% specificity as compared to RPT, i.e., no false-negative results were obtained. Few suspicious samples, which were negative in the RPT after retesting, provided divergent positive results suggesting a lower limit of detection of MAT. MAT is thus able to detect contaminants in biological products such as HS batches.

Entomopathogenic potential of Metarhizium anisopliae isolated from engorged females and tested in eggs and larvae of Boophilus microplus (Acari: Ixodidae).

The purpose of this work was to evaluate the in vitro virulence of three isolates of Metarhizium anisopliae var. anisopliae to eggs and larvae of the tick Boophilus microplus. The fungus tested was isolated from engorged females of B. microplus collected in the field, and identified as Ma01, Ma02 e Ma04. These isolates were evaluated by immersion of eggs and larvae in suspension with different conidial concentrations: 10(5), 10(6), 10(7) e 10(8) conidia/ml. In each isolate there was a treatment group for each spore concentration and a control group with 10 repetitions. It was observed in the treated egg groups that there was a hatching percentage that was much less than that observed in the control groups. This was in inverse proportion to the conidia concentration/ml. Larval bioassays of all the tested isolates resulted in a high mortality of larvae in direct proportion to the spore concentration/ml, 10 days after the conidia suspensions were inoculated. To consolidate the infection, the fungus used in bioassays was re-isolated.

Beauveria bassiana isolated from engorged females and tested against eggs and larvae of Boophilus microplus (Acari: Ixodidae).

The purpose of this work was to evaluate the in vitro virulence of three isolates of Beauveria bassiana to eggs and larvae of the tick Boophilus microplus. The fungus tested were isolated from engorged females of B. microplus collected on the field, and identified as Bb28, Bb29 and Bb30. These isolates were evaluated by immersion of eggs and larvae in suspensions with different conidial concentrations: 10(8), 10(7), 10(6) and 10(5) conidia/ml and compared to the control groups. In the treated eggs, there was a percentage much smaller of hatching than that observed in the controls. The egg hatch was inversely proportional to the conidial concentration. In larval bioassays, all isolates resulted in a higher mortality of larvae compared to the control according to the conidial concentrations/ml, 10 days after treatment inoculation.

Entomopathogenic effect of Aspergillus giganteus and Penicillium corylophilum on two triatomine vectors of Chagas disease.

Two strains, Penicillium corylophilum and Aspergillus giganteus, of the most frequent species found in a survey of triatomines, were used for bioassays in the second and fourth nymphs stage of Triatoma infestans and Panstrongylus megistus. Two procedures, bite and pulverization, were used and compared. A. giganteus was most effective, causing mortality in at least 50% of the nymphs of the two species tested with exception of the nymphs of the fourth stage of P. megistus. Variation in entomopathogenic capacity of the fungal species were observed in the experiments. The two procedures used proved effective.