Effects of amino acid levels during rearing on Cobb 500 slow-feathering broiler breeders: 1. Growth and development.

Adequate pullet nutrition is essential to reach BW and suitable body composition for reproduction. An experiment was conducted to determine the effects of 4 dietary amino acid (AA) levels on BW, flock uniformity, body conformation, organ, leg, and feathering development of broiler breeder pullets during the rearing phase from 5 to 24 wk. A total of 1,360 Cobb-500 slow-feathering (SF) pullets were randomly placed in 16-floor pens with 85 females per pen. Diets with corn, soybean meal, and wheat-midds were formulated without protein restriction maintaining minimum ratios between essential AA and Lys on a digestible (dig) ideal basis. Treatments consisted of 4 dietary AA levels with 80% (low-AA), 90% (moderate-AA), 100% (standard-AA), and 110% (high-AA) of the Cobb-Vantress recommendations guided by dig Lys using balanced protein. Up to 4 wk, all pullets were fed a common starter crumble diet. Grower and developer mash diets were fed to pullets from 5 to 15 wk and from 16 to 24 wk, respectively. Pullets fed standard-AA and high-AA diets were heavier (P < 0.001) than those fed low-AA diets at 10, 15, and 20 wk of age. High-AA diets resulted in better (P = 0.040) flock uniformity at 10 wk. Pullets fed a high-AA diet had the highest (P = 0.041) relative breast weight at 20 wk of age and the lowest (P = 0.044) deposits of abdominal fat at 15 wk of age. Fleshing increased (P < 0.05) as AA content rise in the diet, while the relative shank length (P < 0.001) and the number of wing juvenile feathers (P = 0.004) decreased. Pullets fed the lowest dietary AA level had the longest (P = 0.007) small intestine relative to BW at 10 wk of age, but a smaller (P = 0.001) liver than those fed moderate and standard-AA diets at 20 wk of age. Dietary AA levels have important effects on pullet BW, fleshing, and organ development during rearing with potential reproductive performance impacts.

Optimized DNA extraction and purification method for characterization of bacterial and fungal communities in lung tissue samples.

Human lungs harbor a scarce microbial community, requiring to develop methods to enhance the recovery of nucleic acids from bacteria and fungi, leading to a more efficient analysis of the lung tissue microbiota. Here we describe five extraction protocols including pre-treatment, bead-beating and/or Phenol:Chloroform:Isoamyl alcohol steps, applied to lung tissue samples from autopsied individuals. The resulting total DNA yield and quality, bacterial and fungal DNA amount and the microbial community structure were analyzed by qPCR and Illumina sequencing of bacterial 16S rRNA and fungal ITS genes. Bioinformatic modeling revealed that a large part of microbiome from lung tissue is composed of microbial contaminants, although our controls clustered separately from biological samples. After removal of contaminant sequences, the effects of extraction protocols on the microbiota were assessed. The major differences among samples could be attributed to inter-individual variations rather than DNA extraction protocols. However, inclusion of the bead-beater and Phenol:Chloroform:Isoamyl alcohol steps resulted in changes in the relative abundance of some bacterial/fungal taxa. Furthermore, inclusion of a pre-treatment step increased microbial DNA concentration but not diversity and it may contribute to eliminate DNA fragments from dead microorganisms in lung tissue samples, making the microbial profile closer to the actual one.