Identification of fungi of the genus Aspergillus section nigri using polyphasic taxonomy

In spite of the taxonomy of the Aspergillus species of the Nigri Section being regarded as troublesome, a number of methods have been proposed to aid in the classification of this Section. This work aimed to distinguish Aspergillus species of the Nigri Section from foods, grains and caves on the basis in Polyphasic Taxonomy by utilizing morphologic and physiologic characters, and sequencing of ß-tubulin and calmodulin genes. The morphologic identification proved useful for some species, such as A. carbonarius and Aspergillus sp UFLA DCA 01, despite not having been totally effective in elucidating species related to A. niger. The isolation of the species of the Nigri Section on Creatine Sucrose Agar (CREA) enabled to distinguish the Aspergillus sp species, which was characterized by the lack of sporulation and by the production of sclerotia. Scanning Electron microscopy (SEM) allowed distinguishing the species into two distinct groups. The production of Ochratoxin A (OTA) was only found in the A. carbonarius and A. niger species. The sequencing of β-tubulin gene was efficient in differing most of the Aspergillus species from the Nigri Section with the exception of Aspergillus UFLA DCA 01, which could not be distinguished from A. costaricaensis. This species is morphologically similar to A. costaricaencis for its low sporulation capacity and high sclerotia production, but it differs morphologically from A. costaricaensis for its conidial ornamentation and size of vesicles. Equally, based on partial calmodulin gene sequence data Aspergillus UFLA DCA 01 differs from A. costaricaensis.

Phylogenetic position of an uncharacterized Brazilian strain of bovine papillomavirus in the genus Xipapillomavirus based on sequencing of the L1 open reading frame

The use of PCR assays with degenerate primers has suggested the existence of numerous as yet uncharacterized bovine papillomaviruses (BPV). Despite the endemic nature of BPV infections, the identification of BPV types in Brazilian cattle is still only sporadic. However, in a recent analysis of a partial segment of the L1 gene, we observed notable diversity among the BPV types detected. The aim of this study was to determine the phylogenetic position of the previously identified wild strain BPV/BR-UEL2 detected in the state of Paraná in Brazil. Since previous analysis of the partial L1 sequence had shown that this strain was most closely related to BPV type 4, genus-specific primers were designed. Phylogenetic analysis using complete L1 ORF sequences revealed that BPV/BR-UEL2 was related to BPV types classified in the genus Xipapillomavirus and shared the highest L1 nucleotide sequence similarity with BPV type 4 (78 percent). This finding suggests that BPV/BR-UEL2 should be classified as a potential new type of BPV in the genus Xipapillomavirus.

Identification of the recently described new type of bovine papillomavirus (BPV-8) in a Brazilian beef cattle herd: [revision] / Identificação do novo tipo de papilomavírus bovino (BPV-8) recentemente descrito num rebanho bovino no Brasil: [revisão]

Bovine papillomavirus type 8 (BPV-8) was first detected and described in teat warts as well as in healthy teat skin from cattle raised in Japan. The entire viral genome was sequenced in 2007. Additionally, a variant of BPV-8, BPV-8-EB, was also identified from papillomatous lesions of a European bison in Slovakia. In Brazil, despite the relatively common occurrence of BPV infections, the identification and determination of viral types present in cattle is still sporadic. The aim of this study is to report the occurrence of the recently described BPV-8 in Brazil. The virus was identified in a skin warts obtained from a beef cattle herd located in Parana state, southern Brazil. The papilloma had a macular, non-verrucous gross aspect and was located on the dorsal thorax of a cow. Polymerase chain reaction (PCR) was performed using generic primers for partial amplification of L1 gene. The obtained amplicon (480bp) was cloned and two selected clones were sequenced. The nucleotide sequence was compared to existing papillomaviral genomic sequences, identifying the virus as BPV type 8. This study represents the first report of BPV-8 occurrence in Brazil, what suggests its presence among Brazilian cattle.

A primeira descrição do papilomavírus bovino tipo 8 (BPV-8) foi realizada em amostras de papilomas de teto e de pele saudável de tetos de bovinos no Japão. Em 2007, a seqüência genômica completa do BPV-8 foi determinada. Ainda em 2007, uma variante do BPV-8 (BPV-8-EB) foi identificada em lesões papilomatosas de um bisão europeu na Eslováquia. No Brasil, apesar da infecção pelo BPV ser comumente observada em bovinos, a determinação dos tipos virais associados com a infecção ainda é esporádica. Este estudo tem o objetivo de relatar a ocorrência do BPV-8 no país. A amostra clínica foi obtida em um rebanho de corte do estado do Paraná, região sul do Brasil. O papiloma cutâneo, de aspecto macular e não-verrucoso, estava localizado na região dorsal torácica do animal. A identificação do vírus foi realizada pela reação em cadeia da polimerase (PCR) utilizando primers genéricos para a amplificação parcial do gene L1. O produto amplificado, com aproximadamente 480 pb, foi clonado e os fragmentos presentes em dois clones foram seqüenciados. A comparação da seqüência de nucleotídeos com a de outros papilomavírus demonstrou 100 por cento de identidade com o BPV-8. Assim, esta é a primeira descrição da ocorrência do BPV-8 no Brasil, o que sugere a sua presença nos rebanhos bovinos brasileiros.

Structural characterization of the bglH gene encoding a beta-glucosidase-like enzyme in an endophytic Bacillus pumilus strain

A beta-glucosidase-like enzyme-encoding gene (bglH) of an endophytic Bacillus pumilus strain (CL16) was cloned using a shotgun genomic library constructed in Escherichia coli. The nucleotide sequence of the entire cloned fragment (2484 bp) was determined and characterized. An incomplete open reading frame (ORF) of 534 bp (ORF1) designated bglP and a complete ORF of 1419 bp (ORF2) designated bglH, located in the fragment, are organized in an operon. The protein deduced from 1419 bp (ORF2) had 472 amino acid residues without a characteristic signal peptide sequence, suggesting that the enzyme is localized in the cytoplasm. The amino acid sequence deduced from bglH gene had high similarity with beta-glucosidases from the glycosyl hydrolase family 1. Over-expression of the B. pumilus bglH gene in E. coli showed a 54 kDa protein whose identity was confirmed by mass spectrometry (MALDI-TOF).

Recombination of lac- mal+ cells with lac- mal- variants arisen spontaneously from different Candida albicans strains

Cepas de Candida albicans, mantidas em meios de cultura usuais, originaram colônias que em testes de assimilaçäo de fontes de carbono mostraram-se lac+ mal-. Esferosplastos obtidos a partir de células normais, lac- mal+, foram submetidos à fusäo e produziram apenas células lac+ mal+. Após reduçäo na ploidia,essas células deram origem a colônias lac- mal+ (82 por cento), lac- mal (13,3 por cento) e lac+ mal+ (4,4 por cento). Discute-se neste trabalho, ploidia dos produtos de fusäo, ocorrência de heterocariose nesses produtos e a perda de cromossomos durante a segregaçäo

Resíduos de mandioca induzem uma lata produçäo de enzimas celuloliticas / Cassava residues induce high production of cellulolytic enzymes

O objetivo deste trabalho foi verificar a produçäo de celulase a partir do uso de dois resíduos de procesamento da mandioca (a casca e o farelo). Esta produçäo foi comparada com a produçäo em farelo de trigo, um resíduo tradicionalmente usado pelas indústrias na produçäo enzimas. Os resultados mostraram que a casca de mandioca induziu cerca de 40 (por cento) maior produçäo de celulase (FPA) do que o farelo de trigo (au)