Identification of a Novel Lipase with AHSMG Pentapeptide in Hypocreales and Glomerellales Filamentous Fungi.

Lipases are enzymes that hydrolyze triglycerides to fatty acids and glycerol. A typical element in lipases is a conserved motif of five amino acids (the pentapeptide), most commonly G-X-S-X-G. Lipases with the pentapeptide A-X-S-X-G are present in species of Bacillus, Paucimonas lemoignei, and the yeast Trichosporon asahii; they are usually thermotolerant and solvent resistant. Recently, while searching for true lipases in the Trichoderma harzianum genome, one lipase containing the pentapeptide AHSMG was identified. In this study, we cloned from T. harzianum strain B13-1 the lipase ID135964, renamed here as ThaL, which is 97.65% identical with the reference. We found that ThaL is a lid-containing true lipase of cluster III that belongs to a large family comprising highly conserved proteins in filamentous fungi in the orders Hypocreales and Glomerellales, in which predominantly pathogenic fungi are found. ThaL was expressed in conidia, as well as in T. harzianum mycelium, where it was cultured in liquid minimal medium. These results-together with the amino acid composition, absence of a signal peptide, mitochondrial sorting prediction, disordered regions in the protein, and lineage-specific phylogenetic distribution of its homologs-suggest that ThaL is a non-canonical effector. In summary, AHSMG-lipase is a novel lipase family in filamentous fungi, and is probably involved in pathogenicity.

Comparative genomics of plant pathogenic Diaporthe species and transcriptomics of Diaporthe caulivora during host infection reveal insights into pathogenic strategies of the genus.

BACKGROUND: Diaporthe caulivora is a fungal pathogen causing stem canker in soybean worldwide. The generation of genomic and transcriptomic information of this ascomycete, together with a comparative genomic approach with other pathogens of this genus, will contribute to get insights into the molecular basis of pathogenicity strategies used by D. caulivora and other Diaporthe species. RESULTS: In the present work, the nuclear genome of D. caulivora isolate (D57) was resolved, and a comprehensive annotation based on gene expression and genomic analysis is provided. Diaporthe caulivora D57 has an estimated size of 57,86 Mb and contains 18,385 predicted protein-coding genes, from which 1501 encode predicted secreted proteins. A large array of D. caulivora genes encoding secreted pathogenicity-related proteins was identified, including carbohydrate-active enzymes (CAZymes), necrosis-inducing proteins, oxidoreductases, proteases and effector candidates. Comparative genomics with other plant pathogenic Diaporthe species revealed a core secretome present in all Diaporthe species as well as Diaporthe-specific and D. caulivora-specific secreted proteins. Transcriptional profiling during early soybean infection stages showed differential expression of 2659 D. caulivora genes. Expression patterns of upregulated genes and gene ontology enrichment analysis revealed that host infection strategies depends on plant cell wall degradation and modification, detoxification of compounds, transporter activities and toxin production. Increased expression of effectors candidates suggests that D. caulivora pathogenicity also rely on plant defense evasion. A high proportion of the upregulated genes correspond to the core secretome and are represented in the pathogen-host interaction (PHI) database, which is consistent with their potential roles in pathogenic strategies of the genus Diaporthe. CONCLUSIONS: Our findings give novel and relevant insights into the molecular traits involved in pathogenicity of D. caulivora towards soybean plants. Some of these traits are in common with other Diaporthe pathogens with different host specificity, while others are species-specific. Our analyses also highlight the importance to have a deeper understanding of pathogenicity functions among Diaporthe pathogens and their interference with plant defense activation.

Identification and Comparison of Colletotrichum Secreted Effector Candidates Reveal Two Independent Lineages Pathogenic to Soybean.

Colletotrichum is one of the most important plant pathogenic genus of fungi due to its scientific and economic impact. A wide range of hosts can be infected by Colletotrichum spp., which causes losses in crops of major importance worldwide, such as soybean. Soybean anthracnose is mainly caused by C. truncatum, but other species have been identified at an increasing rate during the last decade, becoming one of the most important limiting factors to soybean production in several regions. To gain a better understanding of the evolutionary origin of soybean anthracnose, we compared the repertoire of effector candidates of four Colletotrichum species pathogenic to soybean and eight species not pathogenic. Our results show that the four species infecting soybean belong to two lineages and do not share any effector candidates. These results strongly suggest that two Colletotrichum lineages have acquired the capability to infect soybean independently. This study also provides, for each lineage, a set of candidate effectors encoding genes that may have important roles in pathogenicity towards soybean offering a new resource useful for further research on soybean anthracnose management.

Genome-wide analysis of Claviceps paspali: insights into the secretome of the main species causing ergot disease in Paspalum spp.

BACKGROUND: The phytopatogen Claviceps paspali is the causal agent of Ergot disease in Paspalum spp., which includes highly productive forage grasses such as P. dilatatum. This disease impacts dairy and beef production by affecting seed quality and producing mycotoxins that can affect performance in feeding animals. The molecular basis of pathogenicity of C. paspali remains unknown, which makes it more difficult to find solutions for this problem. Secreted proteins are related to fungi virulence and can manipulate plant immunity acting on different subcellular localizations. Therefore, identifying and characterizing secreted proteins in phytopathogenic fungi will provide a better understanding of how they overcome host defense and cause disease. The aim of this work is to analyze the whole genome sequences of three C. paspali isolates to obtain a comparative genome characterization based on possible secreted proteins and pathogenicity factors present in their genome. In planta RNA-seq analysis at an early stage of the interaction of C. paspali with P. dilatatum stigmas was also conducted in order to determine possible secreted proteins expressed in the infection process. RESULTS: C. paspali isolates had compact genomes and secretome which accounted for 4.6-4.9% of the predicted proteomes. More than 50% of the predicted secretome had no homology to known proteins. RNA-Seq revealed that three protein-coding genes predicted as secreted have mayor expression changes during 1 dpi vs 4 dpi. Also, three of the first 10 highly expressed genes in both time points were predicted as effector-like. CAZyme-like proteins were found in the predicted secretome and the most abundant family could be associated to pectine degradation. Based on this, pectine could be a main component affected by the cell wall degrading enzymes of C. paspali. CONCLUSIONS: Based on predictions from DNA sequence and RNA-seq, unique probable secreted proteins and probable pathogenicity factors were identified in C. paspali isolates. This information opens new avenues in the study of the biology of this fungus and how it modulates the interaction with its host. Knowledge of the diversity of the secretome and putative pathogenicity genes should facilitate future research in disease management of Claviceps spp.

Salmonella Minnesota de origem avícola apresenta fatores de virulência e risco potencial aos humanos / [Salmonella Minnesota poultry origin has virulence factors and potential risk to human]

Objetivou-se avaliar características de virulência, perfil de resistência antimicrobiana e padrão de similaridade genética de 71 cepas de Salmonella Minnesota isoladas na cadeia produtiva de frangos de corte, entre 2009 e 2010, em duas unidades de uma empresa (A e B). Os isolados foram sorotipificados e submetidos ao teste de susceptibilidade antimicrobiana pelo teste de difusão em disco. Utilizando-se PCR, foi avaliada a presença dos genes invA, lpfA, agfA e sefA e os genes de resistência aos betalactâmicos (bla TEM , bla SHV e bla CTX-M ). A relação filogenética foi determinada por RAPD-PCR. Os maiores percentuais de resistência foram para tetraciclina e sulfonamida. Foram reconhecidos oito perfis de resistência aos antimicrobianos entre as cepas isoladas na indústria A, e 11 perfis de resistência na indústria B. Do total de cepas, 100% foram positivas para o gene invA, 98,6% para o gene agfA, 49,3% para o gene lpfA e nenhuma para o gene sefA. Três cepas foram positivas para o gene bla TEM (4,2%) e 11 (15,5%) para o gene bla CTX-M . A avaliação filogenética demonstrou a presença de sete clusters com similaridade superior a 80% e três perfis distintos. Com base no dendrograma, observou-se a disseminação de um mesmo perfil em ambas as empresas.(AU)

The aim of this study was to evaluate virulence characteristics, antimicrobial resistance profile and the pattern of genetic similarity of 71 strains of Salmonella Minnesota isolated in the production chain of broilers between 2009 and 2010, into two units of a company (A and B). Isolates were serotyped and submitted to antimicrobial susceptibility by disk diffusion test. Using PCR, the presence of genes invA, lpfA, agfA and sefA and the genes conferring resistance to beta-lactam (blaTEM, blaSHV and blaCTX-M) were evaluated. The phylogenetic relationship was determined by the RAPD-PCR method. The highest percentages of resistance were to tetracycline and sulfonamide. Eight antimicrobial resistance profiles were recognized among strains isolated in industry A, and 11 resistance profiles in industry B. Of all strains of both industries, 100% were positive for the invA gene, 98.6% to agfA gene, 49.3% for lpfA gene, and no strain showed the sefA gene. Three strains were positive for the gene blaTEM (4.2%), 11 (15.5%) for the blaCTX-M gene. Phylogenetic evaluation showed the presence of seven clusters with similarity greater than 80% and three distinct profiles. Based on the dendrogram we observed the spread with similar profiles in both companies.(AU)

Salmonella Minnesota de origem avícola apresenta fatores de virulência e risco potencial aos humanos / [Salmonella Minnesota poultry origin has virulence factors and potential risk to human]

Objetivou-se avaliar características de virulência, perfil de resistência antimicrobiana e padrão de similaridade genética de 71 cepas de Salmonella Minnesota isoladas na cadeia produtiva de frangos de corte, entre 2009 e 2010, em duas unidades de uma empresa (A e B). Os isolados foram sorotipificados e submetidos ao teste de susceptibilidade antimicrobiana pelo teste de difusão em disco. Utilizando-se PCR, foi avaliada a presença dos genes invA, lpfA, agfA e sefA e os genes de resistência aos betalactâmicos (bla TEM , bla SHV e bla CTX-M ). A relação filogenética foi determinada por RAPD-PCR. Os maiores percentuais de resistência foram para tetraciclina e sulfonamida. Foram reconhecidos oito perfis de resistência aos antimicrobianos entre as cepas isoladas na indústria A, e 11 perfis de resistência na indústria B. Do total de cepas, 100% foram positivas para o gene invA, 98,6% para o gene agfA, 49,3% para o gene lpfA e nenhuma para o gene sefA. Três cepas foram positivas para o gene bla TEM (4,2%) e 11 (15,5%) para o gene bla CTX-M . A avaliação filogenética demonstrou a presença de sete clusters com similaridade superior a 80% e três perfis distintos. Com base no dendrograma, observou-se a disseminação de um mesmo perfil em ambas as empresas.(AU)

The aim of this study was to evaluate virulence characteristics, antimicrobial resistance profile and the pattern of genetic similarity of 71 strains of Salmonella Minnesota isolated in the production chain of broilers between 2009 and 2010, into two units of a company (A and B). Isolates were serotyped and submitted to antimicrobial susceptibility by disk diffusion test. Using PCR, the presence of genes invA, lpfA, agfA and sefA and the genes conferring resistance to beta-lactam (blaTEM, blaSHV and blaCTX-M) were evaluated. The phylogenetic relationship was determined by the RAPD-PCR method. The highest percentages of resistance were to tetracycline and sulfonamide. Eight antimicrobial resistance profiles were recognized among strains isolated in industry A, and 11 resistance profiles in industry B. Of all strains of both industries, 100% were positive for the invA gene, 98.6% to agfA gene, 49.3% for lpfA gene, and no strain showed the sefA gene. Three strains were positive for the gene blaTEM (4.2%), 11 (15.5%) for the blaCTX-M gene. Phylogenetic evaluation showed the presence of seven clusters with similarity greater than 80% and three distinct profiles. Based on the dendrogram we observed the spread with similar profiles in both companies.(AU)

Virulence Factors of the Entomopathogenic Genus Metarhizium.

The fungal genus Metarhizium has been used as an entomopathogen worldwide for approximately 140 years, and its mechanism of infection and its virulence factors have been studied. The present review is a compilation of virulence factors described in the literature to date and their participation in specific stages of the infection process.

The cell wall proteome from two strains of Pseudocercospora fijiensis with differences in virulence.

Pseudocercospora fijiensis causes black Sigatoka disease, the most important threat to banana. The cell wall is crucial for fungal biological processes, including pathogenesis. Here, we performed cell wall proteomics analyses of two P. fijiensis strains, the highly virulent Oz2b, and the less virulent C1233 strains. Strains were starved from nitrogen to mimic the host environment. Interestingly, in vitro cultures of the C1233 strain grew faster than Oz2b in PDB medium, suggesting that C1233 survives outside the host better than the highly virulent Oz2b strain. Both strains were submitted to nitrogen starvation and the cell wall proteins were isolated and subjected to nano-HPLC-MS/MS. A total of 2686 proteins were obtained from which only 240 had a known function and thus, bioinformatics analyses were performed on this group. We found that 90 cell wall proteins were shared by both strains, 21 were unique for Oz2b and 39 for C1233. Shared proteins comprised 24 pathogenicity factors, including Avr4 and Ecp6, two effectors from P. fijiensis, while the unique proteins comprised 16 virulence factors in C1233 and 11 in Oz2b. The P. fijiensis cell wall proteome comprised canonical proteins, but thirty percent were atypical, a feature which in other phytopathogens has been interpreted as contamination. However, a comparison with the identities of atypical proteins in other reports suggests that the P. fijiensis proteins we detected were not contaminants. This is the first proteomics analysis of the P. fijiensis cell wall and our results expands the understanding of the fundamental biology of fungal phytopathogens and will help to decipher the molecular mechanisms of pathogenesis and virulence in P. fijiensis.

Genome sequence and effectorome of Moniliophthora perniciosa and Moniliophthora roreri subpopulations.

BACKGROUND: The hemibiotrophic pathogens Moniliophthora perniciosa (witches' broom disease) and Moniliophthora roreri (frosty pod rot disease) are among the most important pathogens of cacao. Moniliophthora perniciosa has a broad host range and infects a variety of meristematic tissues in cacao plants, whereas M. roreri infects only pods of Theobroma and Herrania genera. Comparative pathogenomics of these fungi is essential to understand Moniliophthora infection strategies, therefore the detection and in silico functional characterization of effector candidates are important steps to gain insight on their pathogenicity. RESULTS: Candidate secreted effector proteins repertoire were predicted using the genomes of five representative isolates of M. perniciosa subpopulations (three from cacao and two from solanaceous hosts), and one representative isolate of M. roreri from Peru. Many putative effectors candidates were identified in M. perniciosa: 157 and 134 in cacao isolates from Bahia, Brazil; 109 in cacao isolate from Ecuador, 92 and 80 in wild solanaceous isolates from Minas Gerais (Lobeira) and Bahia (Caiçara), Brazil; respectively. Moniliophthora roreri showed the highest number of effector candidates, a total of 243. A set of eight core effectors were shared among all Moniliophthora isolates, while others were shared either between the wild solanaceous isolates or among cacao isolates. Mostly, candidate effectors of M. perniciosa were shared among the isolates, whereas in M. roreri nearly 50% were exclusive to the specie. In addition, a large number of cell wall-degrading enzymes characteristic of hemibiotrophic fungi were found. From these, we highlighted the proteins involved in cell wall modification, an enzymatic arsenal that allows the plant pathogens to inhabit environments with oxidative stress, which promotes degradation of plant compounds and facilitates infection. CONCLUSIONS: The present work reports six genomes and provides a database of the putative effectorome of Moniliophthora, a first step towards the understanding of the functional basis of fungal pathogenicity.

Staphylococcus hyicus / Staphylococcus hyicus

Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.(AU)

Staphylococcus hyicus is an important micro organism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the specie's characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.(AU)

Staphylococcus hyicus / Staphylococcus hyicus

Staphylococcus hyicus is an important microorganism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the species characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.

Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.

Staphylococcus hyicus

ABSTRACT:Staphylococcus hyicus is an important micro organism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the specie's characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.

RESUMO:Staphylococcus hyicus é um micro-organismo de impor tância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O pre sente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de pato- genicidade e sua ocorrência em animais, alimentos e seres humanos.

Staphylococcus hyicus / Staphylococcus hyicus

Staphylococcus hyicus is an important microorganism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the species characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.(AU)

Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.(AU)

Staphylococcus hyicus

ABSTRACT:Staphylococcus hyicus is an important micro organism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the specie's characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.

RESUMO:Staphylococcus hyicus é um micro-organismo de impor tância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O pre sente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de pato- genicidade e sua ocorrência em animais, alimentos e seres humanos.

Pathogenic factors and antimicrobial resistance of Staphylococcus epidermidis associated with nosocomial infections occurring in intensive care units

Nosocomial infections constitute an important problem in hospitals, intensive care units (ICU) having the highest incidence of this type of infection. Staphylococci, especially Staphylococcus epidermidis and Staphylococcus aureus, are among the most important microorganisms associated with nosocomial infections. S. epidermidis is a common skin resident, and can be introduced into the clinical environment by patients and hospital staff. The situation in hospitals is aggravated by the emergence of multiresistant strains. We evaluated 98 hospital S. epidermidis isolates collected at neonatal, pediatric and adult ICUs and 20 S. epidermidis control skin resident isolates from healthy volunteers, for resistance to ten antibiotics and chemotherapeutic agents, and other pathogenicity factors. A high frequency (76.5%) of multiresistance was detected in clinical isolates, whereas community isolates were resistant to penicillin and ampicillin only. The frequency of multiresistant strains was 67.7% in the neonatal ICU, 66.6% in the pediatric ICU and 60.8% in the adult ICU, the lower frequency of multiresistant isolates in the adult ICU indicates a higher incidence of community strains in this unit. There were significantly higher frequencies of hemolytic, proteolytic and biofilm-forming isolates in the clinical isolates than the community isolates, indicating a higher incidence of strains with pathogenic potential in the hospital environment. Except for slight correlation with hemolytic activity there was no correlation between antibiotic multiresistance and pathogenicity factors.

As infecções nosocomiais constituem um importante problema em hospitais, sendo que as unidades de terapia intensiva (UTI) apresentam a maior incidência deste tipo de infecções. Os estafilococos, especialmente S. epidermidis e S. aureus, estão entre os microrganismos mais importantes associados às infecções nosocomiais. S. epidermidis é uma espécie colonizante da pele, sendo frequentemente inoculado durante procedimentos invasivos ou veiculado pela equipe de saúde, e essa situação é agravada pela emergência de cepas multirresistentes endêmicas no ambiente hospitalar. No presente trabalho foi avaliada a resistência a antibióticos e presença de distintos fatores de patogenicidade em 98 isolados clínicos de S. epidermidis obtidos de vários materiais em unidades de terapia intensiva e 20 colonizantes de pele de voluntários saudáveis. Os resultados obtidos mostraram elevada freqüência de isolados clínicos multirresistentes a antibióticos (76,5%), enquanto esta não foi detectada em isolados de voluntários saudáveis. A frequência de isolados multiresistentes foi 67,7% na UTI neonatal, 66,6% na UTI pediátrica, e 60,8% na UTI adulto, a menor freqüência de isolados multirresistentes na UTI adulto é indicativo de maior incidência de cepas comunitárias nesse local. As diferenças significativas na freqüência de isolados com atividades hemolítica, proteolítica e formação de biofilme, encontradas entre isolados clínicos e controles, indica a maior incidência de cepas com potencial patogênico no ambiente hospitalar. Não foi observada correlação entre multirresistência e fatores de patogenicidade, exceto uma baixa correlação positiva com a atividade hemolítica.