Quantitative trait loci for bone traits segregating independently of those for growth in an F-2 broiler X layer cross

An F broiler-layer cross was phenotyped for 18 skeletal traits at 6, 7 and 9 weeks of age and genotyped with 120 microsatellite markers. Interval mapping identified 61 suggestive and significant QTL on 16 of the 25 linkage groups for 16 traits. Thirty-six additional QTL were identified when the assumption that QTL were fixed in the grandparent lines was relaxed. QTL with large effects on the lengths of the tarsometatarsus, tibia and femur, and the weights of the tibia and femur were identified on GGA4 between 217 and 249 cM. Six QTL for skeletal traits were identified that did not co-locate with genome wide significant QTL for body weight and two body weight QTL did not coincide with skeletal trait QTL. Significant evidence of imprinting was found in ten of the QTL and QTL x sex interactions were identified for 22 traits. Six alleles from the broiler line for weight- and size-related skeletal QTL were positive. Negative alleles for bone quality traits such as tibial dyschondroplasia, leg bowing and tibia twisting generally originated from the layer line suggesting that the allele inherited from the broiler is more protective than the allele originating from the layer

Search for QTL affecting the shape of the egg laying curve of the Japanese quail

BACKGROUND: Egg production is of critical importance in birds not only for their reproduction but also for human consumption as the egg is a highly nutritive and balanced food. Consequently, laying in poultry has been improved through selection to increase the total number of eggs laid per hen. This number is the cumulative result of the oviposition, a cyclic and repeated process which leads to a pattern over time (the egg laying curve) which can be modelled and described individually. Unlike the total egg number which compounds all variations, the shape of the curve gives information on the different phases of egg laying, and its genetic analysis using molecular markers might contribute to understand better the underlying mechanisms. The purpose of this study was to perform the first QTL search for traits involved in shaping the egg laying curve, in an F(2 )experiment with 359 female Japanese quail. RESULTS: Eight QTL were found on five autosomes, and six of them could be directly associated with egg production traits, although none was significant at the genome-wide level. One of them (on CJA13) had an effect on the first part of the laying curve, before the production peak. Another one (on CJA06) was related to the central part of the curve when laying is maintained at a high level, and the four others (on CJA05, CJA10 and CJA14) acted on the last part of the curve where persistency is determinant. The QTL for the central part of the curve was mapped at the same position on CJA06 than a genome-wide significant QTL for total egg number detected previously in the same F(2). CONCLUSION: Despite its limited scope (number of microsatellites, size of the phenotypic data set), this work has shown that it was possible to use the individual egg laying data collected daily to find new QTL which affect the shape of the egg laying curve. Beyond the present results, this new approach could also be applied to longitudinal traits in other species, like growth and lactation in ruminants, for which good marker coverage of the genome and theoretical models with a biological significance are available

The GTPase Activating Rap/RanGAP Domain-Like 1 Gene Is Associated with Chicken Reproductive Traits

BACKGROUND: Abundant evidence indicates that chicken reproduction is strictly regulated by the hypothalamic-pituitary-gonad (HPG) axis, and the genes included in the HPG axis have been studied extensively. However, the question remains as to whether any other genes outside of the HPG system are involved in regulating chicken reproduction. The present study was aimed to identify, on a genome-wide level, novel genes associated with chicken reproductive traits. METHODOLOGY/PRINCIPAL FINDING: Suppressive subtractive hybridization (SSH), genome-wide association study (GWAS), and gene-centric GWAS were used to identify novel genes underlying chicken reproduction. Single marker-trait association analysis with a large population and allelic frequency spectrum analysis were used to confirm the effects of candidate genes. Using two full-sib Ningdu Sanhuang (NDH) chickens, GARNL1 was identified as a candidate gene involved in chicken broodiness by SSH analysis. Its expression levels in the hypothalamus and pituitary were significantly higher in brooding chickens than in non-brooding chickens. GWAS analysis with a NDH two tail sample showed that 2802 SNPs were significantly associated with egg number at 300 d of age (EN300). Among the 2802 SNPs, 2 SNPs composed a block overlapping the GARNL1 gene. The gene-centric GWAS analysis with another two tail sample of NDH showed that GARNL1 was strongly associated with EN300 and age at first egg (AFE). Single marker-trait association analysis in 1301 female NDH chickens confirmed that variation in this gene was related to EN300 and AFE. The allelic frequency spectrum of the SNP rs15700989 among 5 different populations supported the above associations. Western blotting, RT-PCR, and qPCR were used to analyze alternative splicing of the GARNL1 gene. RT-PCR detected 5 transcripts and revealed that the transcript, which has a 141 bp insertion, was expressed in a tissue-specific manner. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that the GARNL1 gene contributes to chicken reproductive traits

Empirical Evaluation of Bone Extraction Protocols

The application of high-resolution analytical techniques to characterize ancient bone proteins requires clean, efficient extraction to obtain high quality data. Here, we evaluated many different protocols from the literature on ostrich cortical bone and moa cortical bone to evaluate their yield and relative purity using the identification of antibody-antigen complexes on enzyme-linked immunosorbent assay and gel electrophoresis. Moa bone provided an ancient comparison for the effectiveness of bone extraction protocols tested on ostrich bone. For the immunological part of this study, we focused on collagen I, osteocalcin, and hemoglobin because collagen and osteocalcin are the most abundant proteins in the mineralized extracellular matrix and hemoglobin is common in the vasculature. Most of these procedures demineralize the bone first, and then the remaining organics are chemically extracted. We found that the use of hydrochloric acid, rather than ethylenediaminetetraacetic acid, for demineralization resulted in the cleanest extractions because the acid was easily removed. In contrast, the use of ethylenediaminetetraacetic acid resulted in smearing upon electrophoretic separation, possibly indicating these samples were not as pure. The denaturing agents sodium dodecyl sulfate, urea, and guanidine HCl have been used extensively for the solubilization of proteins in non-biomineralized tissue, but only the latter has been used on bone. We show that all three denaturing agents are effective for extracting bone proteins. One additional method tested uses ammonium bicarbonate as a solubilizing buffer that is more appropriate for post-extraction analyses (e.g., proteomics) by removing the need for desalting. We found that both guanidine HCl and ammonium bicarbonate were effective for extracting many bone proteins, resulting in similar electrophoretic patterns. With the increasing use of proteomics, a new generation of scientists are now interested in the study of proteins from not only extant bone but also from ancient bone

NÍVEIS DE CÁLCIO E RELAÇÃO CÁLCIO: FÓSFORO EM RAÇÕES PARA POEDEIRAS LEVES DE 24 A 40 SEMANAS DE IDADE

v. 17, n. 2, p. 206-216, abr./jun. 2016.Objetivou-se determinar a exigência nutricional de cálcio e a melhor relação cálcio:fósforo em rações para poedeiras leves de 24 a 40 semanas de idade. Foram utilizadas 324 aves Hy-Line W-36, distribuídas em delineamento inteiramente casualizado, esquema fatorial 3x3, sendo três níveis de cálcio (3,9; 4,2 e 4,5 %) e três relações cálcio/fósforo (9,3; 10,53 e 12,12/1), totalizando nove tratamentos, seis repetições e seis aves por unidade experimental. Avaliou-se o desempenho produtivo, a qualidade do ovo, as características ósseas, o balanço de cálcio e fósforo e a mortalidade. Não houve interação (P>0,05) entre os níveis de cálcio e as relações cálcio/fósforo para nenhuma das variáveis, porém os níveis de cálcio promoveram efeito (P<0,05) sobre o peso e porcentagem da casca, cálcio na casca, ovos perdidos, consumo de cálcio e fósforo, cálcio e fósforo na excreta, cálcio e fósforo retido. A relação cálcio/fósforo influenciou significativamente (P<0,05) o consumo de ração, ganho de peso, conversão alimentar, consumo de cálcio e fósforo, fósforo na excreta e fósforo retido. Rações contendo 4,5% de cálcio e relação cálcio:fósforo de 12,12:1, correspondendo ao consumo de cálcio de 3,71 g/ave/dia e de fósforo de 306 mg/ave/dia, garantem desempenho satisfatório em rações para poedeiras leves no período de 24 a 40 semanas.The objective of this study was to determine the nutritional requirement for calcium and the best calcium:phosphorus ratio in the diet for laying hens from 24 to 40 weeks of age. 324 Hy-Line W-36 birds were distributed in a completely randomized design in 3x3 factorial scheme, consisting of three levels of calcium (3.9, 4.2, and 4.5%) and three calcium:phosphorus ratios (9.3, 10.53, and 12.12:1) totaling nine treatments, six replicates and six birds each. We evaluated performance, egg quality, bone characteristics, balance of calcium and phosphorus, and mortality. There was no interaction (P> 0.05) between calcium levels and calcium:phosphorus ratio for the variables, but levels of calcium affected (P <0.05) weight and shell percentage, calcium in the shell, lost eggs, consumption of calcium and phosphorus, calcium and phosphorus excretion, and retained calcium and phosphorus. The calcium:phosphorus ratio affected significantly (P <0.05) feed intake, weight gain, feed conversion per dozen eggs, egg mass, calcium and phosphorus intake, phosphorus excretion, and retained phosphorus. Diets containing 4.5% calcium and the calcium:phosphorus ratio of 12,12:1, corresponding to a calcium intake of 3.71 g/hen/day and phosphorus of 306 mg/ bird /day, guarantee satisfactory performance in diets for laying hens from 24 to 40 weeks

Does increased local bone resorption secondary to breast and prostate cancer result in increased cartilage degradation?

<p>Abstract</p> <p>Background</p> <p>Breast and prostate cancer patients often develop lesions of locally high bone turnover, when the primary tumor metastasizes to the bone causing an abnormal high bone resorption at this site. The objective of the present study was to determine whether local increased bone turnover in breast and prostate cancer patients is associated with an increase in cartilage degradation and to test <it>in vitro </it>whether osteoclasts or cathepsin K alone generate CTXII from human bone.</p> <p>Methods</p> <p>The study included 132 breast and prostate cancer patient, where presence of bone metastases was graded according to the Soloway score. Total bone resorption (CTXI_total) and cartilage degradation (CTXII) were determined.</p> <p>Results</p> <p>Breast and prostate cancer patients with bone metastases revealed significant increased levels of CTXI_totalat Soloway scores 1 and higher compared to patients without bone metastases (p < 0.001). CTXII was statistically elevated at score 3 and 4 (p < 0.01). CTXII/CTXI_totalsignificantly decreased at score 3 and 4 (p < 0.001). Levels of CTXI_total, CTXII and CTXII/CTXI_totalchanged +900%, +130%, and -90%, respectively at Soloway score 4 compared to score 0. The <it>in vitro </it>experiments revealed that osteoclasts released CTXI fragments but not CTXII from bone specimens. The same was observed for cathepsin K.</p> <p>Conclusion</p> <p>Data suggest that an uncoupling between bone resorption and cartilage degradation occurs in breast and lung cancer patient.</p