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.

A naturally occurring recombinant isolate of Lettuce mosaic virus

LMV-Common and LMV-Most are two seed-borne types of Lettuce mosaic virus (LMV), genus Potyvirus. LMV-Most, but not LMV-Common, overcomes the resistance afforded to lettuce by two recessive genes, mo1(1) and mo1(2). An RT-PCR-based assay thought to be specific for LMV-Most also amplified LMV-Tn2, previously typified as LMV-Common. The sequence of selected regions along the genome indicated that LMV-Tn2 is a natural recombinant between LMV-Most and LMV-Common isolates, with a putative recombination site located within the P3 coding region. This is the first evidence of a naturally occurring LMV recombinant isolate.

Identification of a gene encoding adaptin-like protein in the Paracoccidioides brasiliensis genome by random amplified polymorphic DNA analysis

Paracoccidioides brasiliensis isolates are not homogeneous in their patterns of pathogenicity in animals and adhesion to epithelial cells. During this investigation, genotypic differences were observed between two samples of P. brasiliensis strain 18 yeast phase (Pbl 8) previously cultured many times, one taken before (Pb18a) and the other after (Pb18b) animal inoculation. Random amplified polymorphic DNA analysis using the primer OPJ4 distinguished Pb18b from Pbl Ba by one 308 bp DNA fragment, which after cloning and sequencing was shown to encode a polypeptide sequence homologous to the protein beta-adaptin. It is suggested, by comparison to other micro-organisms, that this protein might play an important role in the virulence of P. brasiliensis. This result demonstrates the influence of in vitro subculturing on the genotype of this organism.

Classic and molecular study of Giardia duodenalis in children from a daycare center in the region of Presidente Prudente, São Paulo, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Orchid bee baits attracting bees of the genus Megalopta (Hymenoptera, Halictidae) in Bauru region, São Paulo, Brazil: abundance, seasonality, and the importance of odors for dim-light bees

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Secular dynamics and family identification among highly inclined asteroids in the Euphrosyne region

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

The stable archipelago in the region of the Pallas and Hansa dynamical families

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

Dynamical erosion of asteroid groups in the region of the Phocaea family

In a previous paper, the current state of knowledge of the region containing the Phocaea dynamical family was revised. Here, the dynamical evolution and possible origin of the Phocaea dynamical family and asteroid groups in the region are investigated. First, I study the case of asteroids at high eccentricity (e > 0.31). I find that these objects are unstable because of encounters with Mars on time-scales of up to 270 Myr. The minimum time needed by members of the Phocaea classical family to reach the orbital locations of these objects, 370 Myr, can be used to set a lower limit on the age of the Phocaea family.Next, attention is focused on the chaotic layer previously identified near the nu(6) secular resonance border. Using analytical and numerical tools, I find that the presence of the nu(6) secular resonance forces asteroids with vertical bar g-g(6)vertical bar < 2.55 arcsec yr(-1) to reach eccentricities high enough to allow them to experience deep, close encounters with Mars. Results of the analytical model of Yoshikawa and of my numerical simulations fully explain the low-inclination chaotic region found by Carruba.Finally, I investigate the long-term stability of the minor families and clumps identified in the previous paper, with particular emphasis on a clump only identifiable in the domain of proper frequencies (n, g, g - s) around (6246) Komurotoru. I find that while the clumps identified in the space of proper elements quickly disperse when the Yarkovsky effect is considered, the family around (19536) is still observable for time-scales of more than 50 Myr. The (6246) clump, characterized by its interaction with the nu(5) + nu(16) and 2 nu(6) - nu(16) secular resonances, is robust on time-scales of 50 Myr. I confirm that this group may be the first clump ever detected in the frequency domain that can be associated with a real collisional event.

An analysis of the region of the Phocaea dynamical family

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

Identification, characterization and expression analysis of hepcidin gene in sheep

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Characterization and transcriptional analysis of the promoter region of the Duffy blood group, chemokine receptor (DARC) gene in cattle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N(2) until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.

Characterization and transferability of microsatellite markers of the cultivated peanut (Arachis hypogaea)

Background: the genus Arachis includes Arachis hypogaea (cultivated peanut) and wild species that are used in peanut breeding or as forage. Molecular markers have been employed in several studies of this genus, but microsatellite markers have only been used in few investigations. Microsatellites are very informative and are useful to assess genetic variability, analyze mating systems and in genetic mapping. The objectives of this study were to develop A. hypogaea microsatellite loci and to evaluate the transferability of these markers to other Arachis species.Results: Thirteen loci were isolated and characterized using 16 accessions of A. hypogaea. The level of variation found in A. hypogaea using microsatellites was higher than with other markers. Cross-transferability of the markers was also high. Sequencing of the fragments amplified using the primer pair AhII from 17 wild Arachis species showed that almost all wild species had similar repeated sequence to the one observed in A. hypogaea. Sequence data suggested that there is no correlation between taxonomic relationship of a wild species to A. hypogaea and the number of repeats found in its microsatellite loci.Conclusion: These results show that microsatellite primer pairs from A. hypogaea have multiple uses. A higher level of variation among A. hypogaea accessions can be detected using microsatellite markers in comparison to other markers, such as RFLP, RAPD and AFLP. The microsatellite primers of A. hypogaea showed a very high rate of transferability to other species of the genus. These primer pairs provide important tools to evaluate the genetic variability and to assess the mating system in Arachis species.

DNA barcodes identify marine fishes of São Paulo State, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Population biology of spine shrimp Exhippolysmata oplophoroides (Holthuis) (Caridea, Hippolytidae) in a subtropical region, São Paulo, Brazil

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