Secondary aspergillus fumigatus infection associated with coloidal goiter in a Black-masked Lovebird (Agapornis personata)

Aspergillosis is caused by fungus of Aspergillus genus. Is a multifactorial secondary disease and occurs mainly to immunodeficiency. Goiter is the name to non-inflammatory and non-neoplasic thyroid growth which affecting the animal metabolism. In this report we describe a case of aspergillosis and colloidal goiter in a male Black-masked lovebird (Agapornis personata) diagnosed by post mortem exam. The bird was presented for examination due to severe respiratory signs. An initial palliative treatment was performed in order to relieve the symptoms. Despite this, the patient came to die without performing additional ancillary tests. On gross exam, a pulmonary nodule was observed from which we were able to isolate Aspergillus fumigatus on microbial culture. Histological assessment revealed pulmonary aspergilosis and colloid goiter. Based on histopathological and microbiological assessments we conclude that infection probably was secondary to colloid goiter.

Produção e caracterização parcial da xilanase produzida por Aspergillus sp

As xilanases são enzimas que hidrolisam as ligações 1,4-β-xilosídicas da xilana e que possuem potencial biotecnológico em vários processos industriais, como na clarificação de sucos e vinhos, na fabricação de pães, na produção de biocombustíveis e no tratamento das polpas celulósicas. Além disso, os produtos de hidrólise da xilana, xilooligossacarídeos, podem ser utilizados como ingredientes prebióticos, ou seja, ingredientes nutricionais não digeríveis que estimulam seletivamente a proliferação e a atividade de bactérias benéficas do cólon. O uso de enzimas microbianas nas indústrias se dá devido às suas diversas vantagens sobre os métodos químicos e à facilidade de obtenção do microrganismo. Assim, os objetivos deste trabalho foram: produzir e caracterizar parcialmente a xilanase de Aspergillus sp isolado de pó de café utilizado, purificar parcialmente a enzima produzida e analisar os produtos de hidrólise da xilana. O Aspergillus sp mostrou-se bom produtor de xilanase quando o pó de sabugo de milho foi utilizado como fonte de carbono e a xilanase produzida apresentou melhor atividade específica quando o período de cultivo foi de 168 horas. Os processos de precipitação de proteínas e diálise promoveram o aumento da atividade específica do extrato. A xilanase de Aspergillus sp apresentou pH e temperatura de máxima atividade semelhantes aos de produzidas por Aspergillus niger isolados de outras fontes. A hidrólise da xilana produziu xilose e xilooligossacarídeos. Além da xilanase, o fungo revelou ser produtor de outras proteínas que podem ser estudadas em pesquisas futuras.

Determinação das condições ótimas de fermentação para produção de enzimas celulolíticas pelo fungo Aspergillus niger derivado-marinho, utilizando diferentes subprodutos agroindustriais

Pós-graduação em Biotecnologia - IQ

Produção e caracterização parcial da xilanase produzida por Aspergillus sp

As xilanases são enzimas que hidrolisam as ligações 1,4-β-xilosídicas da xilana e que possuem potencial biotecnológico em vários processos industriais, como na clarificação de sucos e vinhos, na fabricação de pães, na produção de biocombustíveis e no tratamento das polpas celulósicas. Além disso, os produtos de hidrólise da xilana, xilooligossacarídeos, podem ser utilizados como ingredientes prebióticos, ou seja, ingredientes nutricionais não digeríveis que estimulam seletivamente a proliferação e a atividade de bactérias benéficas do cólon. O uso de enzimas microbianas nas indústrias se dá devido às suas diversas vantagens sobre os métodos químicos e à facilidade de obtenção do microrganismo. Assim, os objetivos deste trabalho foram: produzir e caracterizar parcialmente a xilanase de Aspergillus sp isolado de pó de café utilizado, purificar parcialmente a enzima produzida e analisar os produtos de hidrólise da xilana. O Aspergillus sp mostrou-se bom produtor de xilanase quando o pó de sabugo de milho foi utilizado como fonte de carbono e a xilanase produzida apresentou melhor atividade específica quando o período de cultivo foi de 168 horas. Os processos de precipitação de proteínas e diálise promoveram o aumento da atividade específica do extrato. A xilanase de Aspergillus sp apresentou pH e temperatura de máxima atividade semelhantes aos de produzidas por Aspergillus niger isolados de outras fontes. A hidrólise da xilana produziu xilose e xilooligossacarídeos. Além da xilanase, o fungo revelou ser produtor de outras proteínas que podem ser estudadas em pesquisas futuras.

Determinação das condições ótimas de fermentação para produção de enzimas celulolíticas pelo fungo Aspergillus niger derivado-marinho, utilizando diferentes subprodutos agroindustriais

Pós-graduação em Biotecnologia - IQ

Marine Fungi Aspergillus sydowii and Trichoderma sp Catalyze the Hydrolysis of Benzyl Glycidyl Ether

Whole cells of the marine fungi Aspergillus sydowii Gc12, Penicillium raistrickii Ce16, P. miczynskii Gc5, and Trichoderma sp. Gc1, isolated from marine sponges of the South Atlantic Ocean (Brazil), have been screened for the enzymatic resolution of (+/-)-2-(benzyloxymethyl)oxirane (benzyl glycidyl ether; 1). Whole cells of A. sydowii Gc12 catalyzed the enzymatic hydrolysis of (R,S)-1 to yield (R)-1 with an enantiomeric excess (ee) of 24-46% and 3-(benzyloxy)propane-1,2-diol (2) with ee values < 10%. In contrast, whole cells of Trichoderma sp. Gc1 afforded (S)-1 with ee values up to 60% and yields up to 39%, together with (R)-2 in 25% yield and an ee of 32%. This is the first published example of the hydrolysis of 1 by whole cells of marine fungi isolated from the South Atlantic Ocean. The hydrolases from the two studied fungi exhibited complementary regioselectivity in opening the epoxide ring of racemic 1, with those of A. sydowii Gc12 showing an (S) preference and those of Trichoderma sp. Gc1 presenting an (R) preference for the substrate.

THE INFLUENCE OF THE 1992-1993 EL NINO ON THE REPRODUCTIVE BIOLOGY OF Scomber japonicus peruanus (JORDAN & HUBB, 1925)

The influence of the 1992-1993 El Nino events on the reproductive behavior of the Scomber japonicus peruanus (Chub mackerel) was studied from samples collected monthly, along the Peruvian coast (3 degrees 23`S-14 degrees 00`S), from January 1990 to December 1993. The monthly variation of the gonadosomatic index and the frequency of the periods of gonad maturation evidenced that the spawning of the species occurred all year long, being more intense in summer. The values of the gonadosomatic index were higher during the occurrence of the 1992-1993 El Nino, while the body weight and gonad weight decreased. Regarding the condition factor, its values decreased in females over 35 cm in fork length.

Growth parameters and mortality rate of the Scomber japonicus peruanus (Jordan & Hubb, 1925) along the Peruvian coast, South Pacific

The growth parameters and the mortality rates of the Scomber japonicus peruanus (Chub mackerel) were studied based on monthly data of frequency of fork length classes obtained from commercial landings off the Peruvian coast from 1996 to 1998. The asymptotic body length and growth rate values obtained by the ELEFAN I (Electronic Length Frequency Analysis) ranged from 40.20 cm to 42.20 cm and from 0.38 to 0.39, respectively. The oscillation amplitude was 0.60; the Winter point values varied from 0.50 to 0.60 and the performance index from 2.79 to 2.84. The total mortality rate of the Chub mackerel obtained by the linearized catch curve oscillated between 1.68 and 3.35. The rate of fishing mortality varied from 1.16 to 2.78 and the exploitation rate from 0.68 to 0.84. The annual rate of natural mortality estimated by the Pauly`s method ranged from 0.52 to 0.53. The results obtained allow us to conclude that the longevity of the Chub mackerel was slightly over seven years.

Aspergillus fumigatus calcineurin interacts with a nucleoside diphosphate kinase

The Ca2+-calcineurin pathway affects virulence and morphogenesis in filamentous fungi. Here, we identified 37 CalA-interacting proteins that interact with the catalytic subunit of calcineurin (CalA) in Aspergillus fumigatus, including the nucleoside diphosphate kinase (SwoH). The in vivo interaction between CalA and SwoH was validated by bimolecular fluorescence complementation. A. fumigatus swoH is an essential gene. Therefore, a temperature-sensitive conditional mutant strain with a point mutation in the active site, SwoH(V83F), was constructed, which demonstrated reduced growth and increased sensitivity to elevated temperatures. The SwoH(V83F) mutation did not cause a loss in virulence in the Galleria mellonella infection model. Taken together these results imply that CalA interacts with SwoH. (C) 2012 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Molecular characterization by amplified fragment length polymorphism and aflatoxin production of Aspergillus flavus isolated from freshly harvested peanut in Brazil

Brazil contributes substantially to the global peanut production, and the state of Sao Paulo is the largest producer in the country. Peanut crops can be contaminated by Aspergillus flavus strains producing aflatoxins, which are highly toxic and carcinogenic. Thus, the production of high-quality peanuts is crucial both for the commercial peanut industry and as a matter of public health. In this study, we used amplified fragment length polymorphism analysis (AFLP) to investigate the genetic variability among A. flavus strains isolated from fresh peanuts harvested in four different regions in the state of Sao Paulo, and to determine whether the molecular genetic profiles correlated with aflatoxin production or sclerotia formation. AFLP analysis generated 78 fragments ranging from 27 to 365 base pairs in length. Thirteen percent were not polymorphic. Genotyping identified twelve groups of A. flavus. On the basis of the polymorphisms identified, similarity between the isolates ranged from 37% to 100%. Of all isolates collected, 91.7% produced aflatoxins and 83.9% produced small sclerotia. Statistical analysis failed to suggest any relationship between the presence of sclerotia and mean levels of aflatoxins B-1 and B-2. Furthermore, a dendrogram based on AFLP data revealed substantial genetic variability among the A. flavus strains, but showed no correlation between dendrogram groups separated by molecular genetic features and production of aflatoxins B-1 or B-2 or the formation of sclerotia.

Strain-specific polyketide synthase genes of Aspergillus niger

In silico comparison of 34 putative pks genes in Aspergillus niger strain CBS 513.88 versus A. niger strain ATCC 1015 genome revealed significant nucleotide identity (>95% covering a minimum of 99% of the gene sequence) for 31 of these genes (approximately 91%). A. niger CBS 513.88 harbors three putative pks genes (An01g01130, An11g05940, and An15g07920), for which nucleotide identity was not found in A. niger ATCC 1015. To compare the results of the in silico analysis with the in vivo situation, experimental data were obtained for a large number of A. niger strains obtained from different substrates and geographical regions. Three putative Os genes that were found to be variable between the two A. niger strains using bioinformatics tools were in fact strain-specific genes based on experimental data. The PCR amplification signals for the An01g01130, An11g05940, and An15g07920 pks genes were detected in only 97%, 71%, and 26% of the strains, respectively. Southern blot analyses confirmed the PCR data. Because one of the strain-specific pits genes (An15g07920) is located in a putative ochratoxin cluster, we focused our investigation on that region. We assessed the ochratoxin production capability of the 119 A. niger strains and found a positive association between the presence of this pia gene and the capability of the respective strain to produce ochratoxin. (C) 2012 Elsevier B.V. All rights reserved.

Characterization of aflatoxigenic and non-aflatoxigenic strains of Aspergillus section Flavi isolated from corn grains of different geographic origins in Brazil

Aflatoxins can cause great economic losses and serious risks to humans and animals health. The largest aflatoxin producers belong to Aspergillus section Flavi and can occur naturally in food commodities. Studies showed that molecular tools as well as the type of sclerotia produced by the strains could be helpful for identification of Aspergillus species and could be correlated with levels of toxin production. The purpose of this work was to characterize the genetic diversity using AFLP technique, the type of sclerotia and the ability of aflatoxin production by isolated strains from corn of different origins in Brazil, and to verify whether qPCR based on aflR and aflP genes is appropriate for estimating the level of aflatoxin production. All the 75 strains were classified as A. flavus and the AFLP technique showed a wide intraspecific variability within them. Regarding sclerotia production, 34% were classified as S and 66% as L type. Among the aflatoxin-producers, 52.8% produced aflatoxin B-1, while 47.2% aflatoxins B-1 and B-2. Statistical analysis showed no correlation between sclerotia production and aflatoxigenicty, and no correlation between the phylogenetic clusters and aflatoxin production. Concerning the relative expression of aflR and aflP, Pearson's correlation test demonstrated low positive correlation between the expression of the aflR and aflP genes and the production of AFB(1) and AFB(2), but showed high positive correlation between aflR and aflP expression. In contrast to the other reference strains, A. oryzae ATCC 7282 showed no amplification of aflR and aflP. The results highlight the need for detection of reliable and reproducible markers with a high positive correlation with aflatoxin production.

Immobilization of a recombinant endo-1,5-arabinanase secreted by Aspergillus nidulans strain A773

An endo-1,5-arabinanase (abnA) encoding gene from Aspergillus niveus was identified, cloned and successfully expressed in Aspergillus nidulans strain A773. Based on amino acid sequence comparison, the 34-kDa enzyme could be assigned to CAZy GH family 43. Characterization of purified recombinant endo-1,5-arabinanase (AbnA) revealed that it is active at a wide pH range (pH 4.0-7.0) and an optimum temperature at 70 degrees C. The immobilization of the AbnA was performed via covalent binding onto agarose-modified supports: glyoxyl iminodiacetic acid-Ni2+, glyoxyl amine, glyoxyl (4% and 10%) and cyanogen bromide activated sepharose. The yield of immobilization was similar on glyoxyl amine and glyoxyl (96%), and higher than glyoxyl iminodiacetic acid-Ni2+ (43%) support. The thermal inactivation of these immobilized preparations showed that the stability of the AbnA immobilized on glyoxyl 4 and 10% was improved by 4.0 and 10.3-fold factor at 70 degrees C. The half-life of glyoxyl 4% derivative at 60 degrees C was >48 h (pH 5), 9 h (pH 7) and 88 min (pH 9). The major hydrolysis product of debranched arabinan or arabinopentaose by glyoxyl agarose-immobilized AbnA was arabinobiose. (C) 2012 Elsevier B.V. All rights reserved.

Molecular characterization of the Aspergillus nidulans fbxA encoding an F-box protein involved in xylanase induction

The filamentous fungus Aspergillus nidulans has been used as a fungal model system to study the regulation of xylanase production. These genes are activated at transcriptional level by the master regulator the transcriptional factor XInR and repressed by carbon catabolite repression (CCR) mediated by the wide-domain repressor CreA. Here, we screened a collection of 42 A. nidulans F-box deletion mutants grown either in xylose or xylan as the single carbon source in the presence of the glucose analog 2-deoxy-D-glucose, aiming to identify mutants that have deregulated xylanase induction. We were able to recognize a null mutant in a gene (fbxA) that has decreased xylanase activity and reduced xInA and xInD mRNA accumulation. The Delta fbxA mutant interacts genetically with creAd-30, creB15, and creC27 mutants. FbxA is a novel protein containing a functional F-box domain that binds to Skp1 from the SCF-type ligase. Blastp analysis suggested that FbxA is a protein exclusive from fungi, without any apparent homologs in higher eukaryotes. Our work emphasizes the importance of the ubiquitination in the A. nidulans xylanase induction and CCR. The identification of FbxA provides another layer of complexity to xylanase induction and CCR phenomena in filamentous fungi. (C) 2011 Elsevier Inc. All rights reserved.

Xylanase and beta-Xylosidase Production by Aspergillus ochraceus: New Perspectives for the Application of Wheat Straw Autohydrolysis Liquor

The xylanase biosynthesis is induced by its substrate-xylan. The high xylan content in some wastes such as wheat residues (wheat bran and wheat straw) makes them accessible and cheap sources of inducers to be mainly applied in great volumes of fermentation, such as those of industrial bioreactors. Thus, in this work, the main proposal was incorporated in the nutrient medium wheat straw particles decomposed to soluble compounds (liquor) through treatment of lignocellulosic materials in autohydrolysis process, as a strategy to increase and undervalue xylanase production by Aspergillus ochraceus. The wheat straw autohydrolysis liquor produced in several conditions was used as a sole carbon source or with wheat bran. The best conditions for xylanase and beta-xylosidase production were observed when A. ochraceus was cultivated with 1% wheat bran added of 10% wheat straw liquor (produced after 15 min of hydrothermal treatment) as carbon source. This substrate was more favorable when compared with xylan, wheat bran, and wheat straw autohydrolysis liquor used separately. The application of this substrate mixture in a stirred tank bioreactor indicated the possibility of scaling up the process to commercial production.