Gisela Lara da Costa

Production of L-asparaginase by filamentous fungi

L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2% proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation.

Study on morphology, pathogenicity, and genetic variability of Beauveria bassiana isolates obtained from Boophilus microplus tick

Fifty isolates of Beauveria bassiana (Balsamo) Vuillemin, 1912 (Ascomycota: Clavicipitaceae) were analyzed by morphology, for their pathogenic potential to Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) larvae, and by Random Amplified Polymorphic DNA-Polymerase Chain Reaction technique. Morphological analysis demonstrated that isolates present characteristics compatible to those described for B. bassiana in the literature. Virulence test demonstrated that all isolates present lethal effect on larvae and that the lethal concentration varies among isolates. The most virulent isolate was the only one obtained from human infection, which was also the only isolate presenting synnemata. The study on genetic variability among the isolates allowed the identification of 23 electrophoretic profiles. The established groupings suggest that most of the isolates obtained from B. microplus of the same locality present low genetic variation. In this way, the data in the present study will contribute to a meticulous characterization of these B. bassiana isolates.

Novel Aminonaphthoquinone Mannich Bases Derived from Lawsone and their Copper(II) Complexes: Synthesis, Characterization and Antibacterial Activity

A series of novel Mannich bases (HL1-HL13) derived from 2-hydroxy-1,4-naphthoquinone (lawsone), substituted benzaldehydes [C6H2R1R2R3C(O)H] and various primary amines (NH2R4, R4 = n-butyl, benzyl, allyl, 2-furfuryl), and their Cu2+ complexes, [Cu(L1)2]-[Cu(L13)2], have been synthesized and fully characterized by analytical and spectroscopic methods. The structures of complexes 1(R1 = R2 = R3 = H; R4 = Bu), 2(R1 = R3 = H; R2 = NO2; R4= Bu) and 7 (R1 = OH; R2 = R3 = H; R4= Bu) were determined by single crystal X-ray diffraction studies. All complexes crystallize in centrosymmetric space groups, with a copper atom in the inversion centre. Two L- coordinate through the naphthalen-2-olate oxygen and secondary amine-N atoms, forming six-membered chelate rings around the copper atom in a trans-N2O2 environment; spectroscopic data confirm that the other complexes exhibit similar molecular arrangement. The antimicrobial activity of all compounds has been tested on seven different strains of bacteria: Bacillus cereus, Bacillus subtilis, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosaand Staphylococcus aureus. In general, Mannich bases were more active than complexes, HL11(R1 = OH; R2 =H; R3 = Me; R4= Bn) and HL13(R1 = OH; R2 = H; R3 = Br; R4= Bn) being the most potent inhibitors. The MIC for the most active compound HL11against S. Coliwas 20 µmol L-1 (8 µg mL-1), better than Chloramphenicol (90 µmol L-1) and well below most values reported for other naphthoquinones.

Penicillium species in mosquitoes from two Brazilian regions

Penicillium species were isolated from 1073 adult mosquitoes and larvae of Aedes spp., Anopheles spp., Culex spp. and Mansonia spp. captured in the northern and southeastern regions of Brazil. In a total of 24 collections, 198 Penicillium strains were identified in 13 species: Penicillium canescens, P. chrysogenum, P. citrinum, P. corylophilum, P. decumbens, P. expansum, P. fellutanum, P. implicatum, P. janthinellum, P. oxalicum, P. purpurogenum, P. viridicatum and P. waksmanii. The total isolation frequency of the Penicillium species in the investigated mosquitoes was: Anopheles spp. (51.5%), Aedes spp. (15.1%), Culex spp. (23.7%) and Mansonia spp. (10.1%). The Penicillium species with the highest incidence in the mosquito genera were: P. corylophilum, P. janthinellum, P. fellutanum and P. waksmanii. The highest number of mosquitoes were captured in the state of Rio de Janeiro, followed by Rond?nia and Minas Gerais.Penicillium species were isolated from 1073 adult mosquitoes and larvae of Aedes spp., Anopheles spp., Culex spp. and Mansonia spp. captured in the northern and southeastern regions of Brazil. In a total of 24 collections, 198 Penicillium strains were identified in 13 species: Penicillium canescens, P. chrysogenum, P. citrinum, P. corylophilum, P. decumbens, P. expansum, P. fellutanum, P. implicatum, P. janthinellum, P. oxalicum, P. purpurogenum, P. viridicatum and P. waksmanii. The total isolation frequency of the Penicillium species in the investigated mosquitoes was: Anopheles spp. (51.5%), Aedes spp. (15.1%), Culex spp. (23.7%) and Mansonia spp. (10.1%). The Penicillium species with the highest incidence in the mosquito genera were: P. corylophilum, P. janthinellum, P. fellutanum and P. waksmanii. The highest number of mosquitoes were captured in the state of Rio de Janeiro, followed by Rondonia and Minas Gerais.

Isolation of fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae) captured at two natural breeding grounds in the municipality of Seropédica, Rio de Janeiro, Brazil

The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides (4.7%), Fusarium sp. (4.7%), Alternaria alternata (11.9%), Curvularia brachyspora (2.4%), Mycelia sterilia (2.4%) and the Mucorales order (2.4%).

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: 105, 106, 107 e 108 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. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

The entomopathogenic potential of Aspergillus spp. in mosquitoes vectors of tropical diseases

Eleven strains of the most frequent Aspergillus species found in a survey of Brazilian mosquitoes collected in the states of Minas Gerais and Rio de Janeiro, Brazil, were used for bioassays in second‐stage larvae of Aedes fluviatilis and Culex quinquefasciatus.Eleven strains of the most frequent Aspergillus species found in a survey of Brazilian mosquitoes collected in the states of Minas Gerais and Rio de Janeiro, Brazil, were used for bioassays in second-stage larvae of Aedes fluviatilis and Culex quinquefasciatus. Aspergillus ochraceus, A. kanagawaensis and one strain of A. sulphureus were most effective, causing mortality in at least 80% of the larvae of the two mosquito species tested. Variations in entomopathogenic capacity were observed in the experiments with strains of A. sulphureus, A. flavus and A. ochraceus.

Fungal flora of the digestive tract of 5 species of triatomines vectors of Trypanosoma cruzi, Chagas 1909.

A study of the mycobiota in the digestive tract of 5 important species of triatomines, Triatoma brasiliensis, T. infestans, T. sordida, T. pseudomaculata and T. vitticeps, was made. The digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated.The genera with the greatest number of species were Penicillium (19 species), Aspergillus (17 species) and Acremonium (5 species) and the most frequent species, in decreasing order, were Penicillium corylophilum, Aspergillus niger, Penicillium fellutanum, Cladosporium herbarum, Penicillium waksmanii, Aspergillus awamori and Paecilomyces variotii. Among the isolated fungi, we found species that are recognized as entomopathogenic and pathogenic for humans and animals.

Pathogenic action of Penicillium species on mosquito vectors of human tropical diseases.

In vitro bioassays were performed in order to assess the pathogenicity of 13 Penicillium strains in 2nd stage larvae of Aedes aegypti, Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus. Mortality rates began in the first 24 hours, ranging from 0 to 100%. P. corylophilum, P. fellutanum, P. implicatum, P. janthinellum, P. viridicatum and P. waksmanii species tested on Aedes aegypti larvae and at different concentrations presented mortality rates from 0 to 6.6%. These species, when tested on Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus larvae, presented a mortality rate of 3.33% to 100%. Susceptibility of Aedes fluviatilis, Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus mosquitoes to the tested agents, turn P. corylophilum and P. janthinellum into candidates for potential use in biological control of vectors.In vitro bioassays were performed in order to assess the pathogenicity of 13 Penicillium strains in 2nd stage larvae of Aedes aegypti, Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus. Mortality rates began in the first 24 hours, ranging from 0 to 100%. P. corylophilum, P. fellutanum, P. implicatum, P. janthinellum, P. viridicatum and P. waksmanii species tested on Aedes aegypti larvae and at different concentrations presented mortality rates from 0 to 6.6%. These species, when tested on Aedes fluviatilis, Anopheles aquasalis and Culex quinquefasciatus larvae, presented a mortality rate of 3.33% to 100%. Susceptibility of Aedes fluviatilis, Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus mosquitoes to the tested agents, turn P. corylophilum and P. janthinellum into candidates for potential use in biological control of vectors.

Avaliação in vitro dos fungos Aspergillus flavus e Penicillium corylophilum em larvas de Musca domestica (Diptera: Muscidade)

The present study was aimed at the evaluation of two isolates of Aspergillus flavus and two isolates of Penicilium corylophilum on third stage larvae of Musca domestica. The samples tested were isolated directly from M. domestica specimens. Third stage larvae were obtained from pupas of a laboratory colony. Four batches with 20 larvae were used for each of conidia dilution tested (105 to 108 conidia/ml). Larvae were dipped for one minute into conidia dilution, being then moved to dishes and kept under controlled environmental conditions. Two control groups were used: one received a treatment with spreading adhesive and the other one with no treatment at all. Larval surviving rates were: 57.50% and 86.25% for groups treated with 108 conidia/ml of A. flavus; and 63.75% and 72.50% for those who had received the same concentration of P. corylophilum conidia. Larval mortality rates were greater for groups that received higher conidia concentrations, and also increased directly proportional to exposition time. Adult hatching rates on treated groups, were affected by fungi treatment. It was concluded that A. flavus and P. corylophilum presented detrimental effects for third stage larvae of M. domestica