Mycoplasma synoviae cell invasion: Elucidation of the Mycoplasma pathogenesis in chicken

Fluorochrome-labelled cells of two field isolates and Mycoplasma synoviae (Ms) were inoculated onto monolayer cultures of fluorochrome-labelled HEp-2 cells and monitored by confocal laser scanning microscopy (CLSM). Ms was detected initially adhered to and subsequently inside the host cells. Between 24 and 48 h of infection, Ms was detected in the perinuclear region, and after 72 h of infection was confirmed by gentamicin invasion assay. High and low passage Ms strains showed no differences in adherence or invasion. The morphology and the actin filaments of the infected HEp-2 cells were preserved throughout the study period. The observed invasion by Ms is consistent with the biology of Mollicutes, and could explain the difficulties in recovering field isolates of the mycoplasma and in controlling the infection in birds even after long-term antibiotic treatment. (C) 2009 Elsevier Ltd. All rights reserved.

Detecting Major Genes Controlling Robustness of Chicken Body Weight Using Double Generalized Linear Models

Detecting both the majors genes that control the phenotypic mean and those controlling phenotypic variance has been raised in quantitative trait loci analysis. In order to mapping both kinds of genes, we applied the idea of the classic Haley-Knott regression to double generalized linear models. We performed both kinds of quantitative trait loci detection for a Red Jungle Fowl x White Leghorn F2 intercross using double generalized linear models. It is shown that double generalized linear model is a proper and efficient approach for localizing variance-controlling genes. We compared two models with or without fixed sex effect and prefer including the sex effect in order to reduce the residual variances. We found that different genes might take effect on the body weight at different time as the chicken grows.

Fine mapping and replication of QTL in outbred chicken advanced intercross lines

Background: Linkage mapping is used to identify genomic regions affecting the expression of complex traits. However, when experimental crosses such as F2 populations or backcrosses are used to map regions containing a Quantitative Trait Locus (QTL), the size of the regions identified remains quite large, i.e. 10 or more Mb. Thus, other experimental strategies are needed to refine the QTL locations. Advanced Intercross Lines (AIL) are produced by repeated intercrossing of F2 animals and successive generations, which decrease linkage disequilibrium in a controlled manner. Although this approach is seen as promising, both to replicate QTL analyses and fine-map QTL, only a few AIL datasets, all originating from inbred founders, have been reported in the literature. Methods: We have produced a nine-generation AIL pedigree (n = 1529) from two outbred chicken lines divergently selected for body weight at eight weeks of age. All animals were weighed at eight weeks of age and genotyped for SNP located in nine genomic regions where significant or suggestive QTL had previously been detected in the F2 population. In parallel, we have developed a novel strategy to analyse the data that uses both genotype and pedigree information of all AIL individuals to replicate the detection of and fine-map QTL affecting juvenile body weight. Results: Five of the nine QTL detected with the original F2 population were confirmed and fine-mapped with the AIL, while for the remaining four, only suggestive evidence of their existence was obtained. All original QTL were confirmed as a single locus, except for one, which split into two linked QTL. Conclusions: Our results indicate that many of the QTL, which are genome-wide significant or suggestive in the analyses of large intercross populations, are true effects that can be replicated and fine-mapped using AIL. Key factors for success are the use of large populations and powerful statistical tools. Moreover, we believe that the statistical methods we have developed to efficiently study outbred AIL populations will increase the number of organisms for which in-depth complex traits can be analyzed.

Quantitative Analysis of Diepoxybutane Damage within the Chicken β-globin Domain

In industrial polymer and synthetic rubber production facilities, workers are exposed to 1,3-butadiene. This compound is converted in vivo to 1,2,3,4-diepoxybutane (DEB) and has been linked to increased incidences of cancer in these individuals. Carcinogenesis has been attributed to formation of DEB induced DNA interstrand cross-links. Previous studies have demonstrated that DEB cross-links deoxyguanosine residues within 5'-GNC sequences in synthetic DNA, in restriction fragments, and in defined sequence nucleosomes. The current study utilized the polymerase chain reaction (PCR) to examine DEB damage frequencies within nuclear genes, found within "open" regions of chromatin, as compared to regions of unexpressed sequence that reside in tightly packed, "closed" chromatin, to more closely model DEB reactivity in vivo. These initial studies have been performed in chicken liver homogenates. Preliminarily, we have found a dose-dependent DEB lesion-forming response within "open" chromatin. DEB appears to have little-to-no effect upon regions of "closed" chromatin.

Mapping of Diepoxybutane Damage in the Cytochrome b Domain of Mitochondrial DNA and the β-globin Domain of Nuclear Chicken DNA Using Quantitative PCR

Diepoxybutane (DEB), a known industrial carcinogen, reacts with DNA primarily at the N7 position of deoxyguanosine residues and creates interstrand cross-links at the sequence 5'-GNC. Since N7-N7 cross-links cause DNA to fragment upon heating, quantative polymerase chain reaction (QPCR) is being used in this experiment to measure the amount of DEB damage (lesion frequency) with three different targets-mitochondrial (unpackaged), open chromatin region, and closed chromatin region. Initial measurements of DEB damage within these three targets were not consistent because the template DNA was not the limiting reagent in the PCR. Follow-up PCR trials using a limiting amount of DNA are still in progress although initial experimentation looks promising. Sequencing of these three targets to confirm the primer targets has only been successfully performed for the closed chromatin target and does not match the sequence from NIH used to design that primer pair. Further sequencing trials need to be conducted on all three targets to assure that a mitochondrial, open chromatin, and closed chromatin region are actually being amplified in this experimental series.

The feeding activity of Colossoma macropomum larvae (tambaqui) in fishponds with water hyacinth (Eichhornia crassipes) fertilizer

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

Crescimento, produtividade e qualidade de raízes de rabanete cultivadas sob diferentes fontes e doses de adubos orgânicos

Avaliou-se o efeito da aplicação de doses de húmus de minhoca e de esterco bovino no crescimento de rabanete (Raphanus sativus L.), cultivar Crimson Giant, de agosto a setembro de 2002. Fez-se semeadura direta do rabanete com desbaste nove dias após, para adequar ao espaçamento de 0,20 x 0,08 m. Utilizou-se delineamento de blocos ao acaso, com sete tratamentos (15; 30 e 45 t ha-1 de húmus de minhoca e esterco bovino curtido, além do controle sem adição de fertilizante), em três repetições. Houve maior acúmulo de massa seca nas folhas em relação aos demais órgãos da planta, com acúmulo contínuo até os 13 dias após a semeadura, com posterior redução, quando ocorreu o desenvolvimento da raiz. As doses e fontes de adubos orgânicos aplicados pouco influenciaram no crescimento e nas produtividades total e comercial de raízes. O húmus de minhoca apresentou a menor porcentagem de raízes rachadas e isoporizadas nas doses de 9,2 e 20,4 t ha-1, respectivamente. A dose de esterco bovino, de 21,3 t ha-1 promoveu a maior porcentagem de raízes rachadas e a de 45 t ha-1 proporcionou redução em relação à de 15 t ha-1.

Produção de repolho em função da aplicação de boro associada a adubo orgânico

Em condições de campo, em solo argiloso com teor médio de boro, foram avaliados os efeitos da adubação com boro, associada à adubação orgânica, na produção de repolho híbrido Kenzan. Cinco doses de B (0; 2,0; 4,0; 6,0 e 8,0 kg ha-1) foram associadas, em esquema fatorial, à ausência e presença (0 e 10 t ha-1) de vermicomposto de esterco bovino. O delineamento experimental foi em blocos casualizados com três repetições e as adubações orgânica e química foram feitas no sulco, por ocasião do transplantio das mudas. O ponto de colheita foi atingido 75 a 90 dias após o transplantio. A adubação com vermicomposto de esterco de bovino elevou a produção em 3,8 t ha-1, o diâmetro das cabeças de repolho em 0,8 cm e o peso das cabeças em 170 g. A produção de repolho aumentou linearmente com a adubação com B e variou de 52,94 t ha-1 a 65,95 t ha-1. A relação entre B no solo e produção foi linear e positiva.

Organic matter fractions and soil fertility under the influence of liming, vermicompost and cattle manure

Os efeitos do vermicomposto de esterco de curral associado à calagem em atributos da fertilidade do solo foram avaliados através de experimento em vasos empregando um Latossolo Vermelho, distrófico, textura média. Cinco doses do vermicomposto (equivalentes a 0; 28; 42; 56 e 70 t ha-1, peso seco) e cinco doses de calcário (visando elevar a saturação por bases a: 20; 30; 40; 50 e 60%) foram combinadas em esquema fatorial, sendo as amostras de solo incubadas por 180 dias. Para comparação entre o vermicomposto e o esterco de curral, amostras do mesmo solo receberam o equivalente a 70 t ha-1 do esterco de curral que originou o vermicomposto e as cinco doses de calcário listadas anteriormente. Através do cálculo do Índice de Eficiência Agronômica, foi verificado que o potencial de fornecimento de K e de Mg pelo esterco é maior do que o do vermicomposto, e que o de P, é semelhante. O vermicomposto aumentou os teores de Ca2+ e de matéria orgânica (MO), os valores de pH em CaCl2 e a CTC a pH 7. Com o aumento das doses de vermicomposto houve diminuição do C-ácidos húmicos e aumento do C-humina e com a calagem o C-total não aumentou mas houve diminuição do C-ácidos húmicos.

Fuzzy modeling to predict chicken egg hatchability in commercial hatchery

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

Heritabilities and genetic correlations for reproductive traits in an F2 reciprocal cross chicken population

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

Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR

The present study was carried out to report the occurrence Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in chicken abattoirs. Samples of feces; feathers; scald, evisceration, and chiller water; and rinse water of non-eviscerated, eviscerated, and chilled carcass were collected from six chicken abattoirs. Salmonella isolates were identified by a multiplex-PCR using three sets of primers targeting the inuA, pefA, and sefA gene sequences from Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. Salmonella spp. was detected in 10% (29/288) of the samples, whereas serovars Enteritidis and Typhimurium were identified in 62% (7/288), respectively. The results indicate the need to improve hygiene and sanitary standards in poultry slaughter lines, besides the education of food handlers and information to consumers. (c) 2006 Elsevier Ltd. All rights reserved.

Survey of chicken abattoir for the presence of Campylobacter jejuni and Campylobacter coli

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

Heat or cold chronic stress affects organ weights and Hsp70 levels in chicken embryos

This study was carried out with the objective of evaluating the effect of heat (38.8 degreesC) or cold (35.8 degreesC) stress on chicken embryo development and tissues Hsp70 levels, after the 13th day of incubation. Embryo weight (percent egg weight), organ weight (percent embryo weight) and Hsp70 levels (ng Hsp70 mug(-1) total protein) in different tissues (liver, breast muscle, heart, lungs, brain and kidney) were studied at the end of incubation. Cold stress induced a lower embryo weight and lower kidney and lungs weights, whereas heart and liver were lighter in heat-stressed embryos. An interaction between temperature and age was obtained only for Hsp70 levels in kidney and heart. Cold-stressed embryos showed higher Hsp70 levels in the brain, lungs and liver; a decrease in brain and breast muscle Hsp70 levels was seen from the 19th to 20th days in control embryos. Hsp70 levels increased with age in kidneys of control embryos and in heart of heat- and cold-stressed embryos. In conclusion, this study showed that chicken embryo organ weights are affected by incubation temperature, and that Hsp70 expression is tissue dependent (higher levels being seen in the brain) being cold-stress more effective in increasing Hsp70 levels in most studied tissues.

Expression of MyoD, myogenin, myostatin and Hsp70 transcripts in chicken embryos submitted to mild cold or heat

The expression of the MyoD, myogenin, myostatin and Hsp70 genes was estimated in chicken embryos submitted to mild cold (36 +/- 0.5degreesC) or heat (44 +/- 0.5degreesC) for 1 h. 2. Marked decreases in MyoD, myogenin and myostatin transcript levels were observed in embryos exposed to high temperature, contrasting to the higher expression of the Hsp70 mRNA detected in heat-stressed embryos. 3. The exposure of chicken embryos to low temperature significantly affected only the abundance of myogenin mRNA. 4. These findings suggest that myogenic proliferation and differentiation events are compromised by variations in environmental temperature during avian embryogenesis. (C) 2003 Elsevier B.V. Ltd. All rights reserved.