A broad-spectrum mer operon in a multi-drug resistant strain of the fish pathogen, Aeromonas salmonicida

Aeromonas salmonicida AS03, a potential fish pathogen, was isolated from Atlantic salmon, Salmo salar, in 2003. This strain was found to be resistant to ≥1000 mM HgCl2 and ≥32 mM phenylmercuric acetate as well as multiple antimicrobials. Mercury (Hg) and antibiotic resistance genes are often located on the same mobile genetic elements, so the genetic determinants of both resistances and the possibility of horizontal gene transfer were examined. Specific PCR primers were used to amplify and sequence distinctive regions of the mer operon. A. salmonicida AS03 was found to have a pDU1358-like broad-spectrum mer operon, containing merB as well as merA, merD, merP, merR and merT, most similar to Klebsiella pneumonaie plasmid pRMH760. To our knowledge, the mer operon has never before been documented in Aeromonas spp. PCR and gene sequencing were used to identify class 1 integron associated antibiotic resistance determinants and the Tet A tetracycline resistance gene. The transposase and resolvase genes of Tn1696 were identified through PCR and sequencing with Tn21 specific PCR primers. We provide phenotypic and genotypic evidence that the mer operon, the aforementioned antibiotic resistances, and the Tn1696 transposition module are located on a single plasmid or conjugative transposon that can be transferred to E. coli DH5α through conjugation in the presence of low level Hg and absence of any antibiotic selective pressure. Additionally, the presence of low-level Hg or chloramphenicol in the mating media was found to stimulate conjugation, significantly increasing the transfer frequency of conjugation above the transfer frequency measured with mating media lacking both antibiotics and Hg. This research demonstrates that mercury indirectly selects for the dissemination of the antibiotic resistance genes of A. salmonicida AS03.

Infectious bronchitis virus: detection and vaccine Strain differentiation by semi-nested RT-PCR

A semi-nested reverse transcription-polymerase chain reaction (Semi-N-RT-PCR) was developed and used to detect the S glycoprotein gene of infectious bronchitis virus (IBV) strains and to discriminate H120 vaccine strain from other strains. Viral RNA was extracted from the allantoic fluid of chicken embryos and from tissues of chickens experimentally infected with different strains of IBV. Amplification and identification of the viral RNA was performed using two sets of primers complementary to a region of the S glycoprotein gene in the Semi-N-RT-PCR assay. The pair of primers used in the first PCR consisted of universal oligonucleotides flanking a more variable region of S1-S2 gene. The second primer pair was used in the Semi-N-RT-PCR and was comprised of one of the primers from the first universal pair together with either another universal internal oligolucleotide or a oligonucleotide sequence specific for the H120 strain of IBV. The universal primers detected all reference IBV strains and field isolates tested herein. The Semi-N-RT-PCR had high sensitivity and specificity, and was able to differentiate the H120 vaccine strain from other reference IBV strains; including M41 strain. All tissue samples collected from chickens experimentally infected with H120 or M41 strains were positive in the semi-nested RT-PCR using universal primers, while only the H120-infected tissue samples were amplified by the set of primers containing the H120-oligonucleotide. In conclusion, the ability of Semi-N-RT-PCR to detect distinct IBV strains and preliminarily discriminate the vaccine strain (H120) closes a diagnostic gap and offers the opportunity to use comprehensive PCR procedures for the IBV diagnosis.

Protection levels of vaccinated pigeons (Columba livia) against a highly pathogenic newcastle disease virus strain

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of Salmonella Enteritidis and Salmonella Gallinarum in birds by using live vaccine candidate containing attenuated Salmonella Gallinarum mutant strain

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic Diversity of a Brazilian Strain Collection of Xanthomonas citri subsp citri Based on the Type III Effector Protein Genes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prevalence of Lettuce mosaic virus - common strain on three lettuce producing areas from São Paulo State

O LMV ocorre em todo o mundo e é considerado um dos patógenos mais importantes para a cultura da alface. de acordo com a habilidade em contornar os genes de resistência mo1¹ e mo1² encontrados em alface, os isolados de LMV podem ser dividos em dois sub-grupos: LMV-Most, capazes de contornar a resistência propiciada por estes genes e de serem transmitidos pela semente nestas cutivares, e LMV-Common, que não são capazes de causar sintomas nestes cultivares, além de serem transmitidos pela semente somente em cultivares suscetíveis. Para avaliar a ocorrência destes dois tipos de isolados de LMV foram coletadas, durante 2002-2005, amostras de alface com sintomas de mosaico em áreas de produção de alface comercial das regiões de Campinas, Mogi das Cruzes e Bauru no estado de São Paulo. O RNA total foi utilizado para detecção por RT-PCR utilizando-se oligonucleotídeos universais para LMV que amplificam a porção N-terminal variável da capa protéica, localizada no terminal 3´do genoma. As amostras positivas foram analisadas por um segundo primer que amplifica um fragmento da região central (CI-VPg) do genoma viral. Um total de 1362 amostras foram avaliadas, tendo sido detectado o LMV em 504 amostras (37,29%). O LMV-Common prevaleceu em variedades suscetíveis (77,3%). O LMV-Most foi encontrado frequentemente associado a variedades portadoras do gene de tolerância mo1¹. Apesar da existência dos LMV-Most capazes de contornar a resistência em alface, estes não predominam em nossa condições.

Investigation of microwave dielectric relaxation solid state process in the antiferroelectric phase of NaNbO3 ceramics

This letter reports microwave dielectric measurements performed in the antiferroelectric phase of NaNbO3 ceramics from 100 to 450 K. Remarkable dielectric relaxation was found within the antiferroelectric phase and in the vicinity of the ferroelectric-antiferroelectric phase transition. Such dielectric relaxation process was associated with relaxations of polar nanoregions with strong relaxor-like characteristic. In addition, the microwave dielectric measurements also revealed an unexpected and unusual anomaly in the relaxation strength, which was related to a disruption of the antiferroelectric order induced by a possible AFE-AFE phase transition. (C) 2004 Elsevier Ltd. All rights reserved.

Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD)

Trichophyton rubrum é um importante agente causal de dermatomicose. Os métodos de tipagem molecular têm sido recentemente desenvolvidos para responder questões sobre epidemiologia e auxiliar no esclarecimento de recidivas, após o tratamento. As seqüências aleatórias 1- (5'-d[GGTGCGGGAA]-3') e 6- (5'-d[CCCGTCAGCA]-3') foram usadas para tipagem molecular deste fungo por RAPD produzindo variabilidade intraespecífica. Cinco padrões foram observados entre os 10 isolados de T. rubrum, com ambas as seqüências. Foi concluído que a análise por RAPD pode ser utilizada para estudos epidemiológicos.

Population dynamics of DENV-1 genotype V in Brazil is characterized by co-circulation and strain/lineage replacement

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Strain differentiation of Trichophyton rubrum by randomly amplified polymorphic DNA and analysis of rDNA nontranscribed spacer

Trichophyton rubrum is the most common pathogen causing dermatophytosis. Molecular strain-typing methods have recently been developed to tackle epidemiological questions and the problem of relapse following treatment. A total of 67 strains of T rubrum were screened for genetic variation by randomly amplified polymorphic DNA (RAPD) analysis, with two primers, 5'-d[GGTGCGGGAA]-3' and 5'-d[CCCGTCAGCA]-3', as well as by subrepeat element analysis of the nontranscribed spacer of rDNA, using the repetitive subelements TRS-1 and TRS-2. A total of 12 individual patterns were recognized with the first primer and 11 with the second. Phylogenetic analysis of the RAPID products showed a high degree of similarity (> 90 %) among the epidemiologically related clinical isolates, while the other strains possessed 60% similarity. Specific amplification of TRS-1 produced three strain-characteristic banding patterns (PCR types); simple patterns representing one copy of TRS-1 and two copies of TRS-2 accounted for around 85 % of all isolates. It is concluded that molecular analysis has important implications for epidemiological studies, and RAPID analysis is especially suitable for molecular typing in T. rubrum.

The semi-synthetic kaurane ent-16 alpha-methoxykauran-19-oic acid induces vascular relaxation and hypotension in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anelastic relaxation due to hydrogen in Ti-35Nb-7Zr-5Ta alloy

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Photo-Induced Dipole Relaxation Current in Natural Amethyst

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Anelastic Relaxation Measurements in Nb-46wt%Ti Alloys with Interstitial Solutes in Solid Solution

Anelastic relaxation measurements were performed in a Nb-46wt%Ti alloy, in the temperature range of 300 to 700 K, using a torsion pendulum operating at an oscillating frequency near 2.0 Hz. The samples were measured in different conditions: cold worked, annealed in ultra-high vacuum and doped with several quantities of nitrogen. The relaxation spectra obtained were resolved into their component peaks, corresponding to the different kinds of interaction of the interstitial solutes with the metallic matrix. The relaxation parameters of each process were calculated using Debye's elementary peaks.