Molting in Salmonella Enteritidis-challenged laying hens fed alfalfa crumbles. II. Fermentation and microbial ecology response.

The objective of this study was to examine microbial population shifts and short-chain fatty acid (SCFA) responses in the gastrointestinal tract of Salmonella Enteritidis-challenged molted and nonmolted hens fed different dietary regimens. Fifteen Salmonella-free Single Comb Leghorn hens (>50 wk old) were assigned to 3 treatment groups of 5 birds each based on diet in 2 trials: 100% alfalfa crumbles (ALC), full-fed (FF, nonmolted) 100% commercial layer ration, and feed withdrawal (FW). A forced molt was induced by either a 12-d alfalfa diet or FW. In all treatment groups, each hen was challenged by crop gavage orally 4 d after molt induction with a 1-mL inoculum containing 10(6) cfu of Salmonella Enteritidis. Fecal and cecal samples (d 4, 6, 8, 11, and necropsy on d 12) were collected postchallenge. Microbial population shifts were evaluated by PCR-based 16S ribosomal RNA gene amplification and denaturing gradient gel electrophoresis, and SCFA concentrations were measured. Total SCFA in fecal and cecal contents for FW molted hens were generally lower (P < or = 0.05) in the later stages of the molt period when compared to ALC and FF treatment groups. The overall trend of SCFA in cecal and fecal samples exhibited similar patterns. In trials 1 and 2, hens molted with ALC diet generally yielded more similar amplicon band patterns with the FF hens in both fecal and cecal samples by the end of the molting period than with FW hens. The results of these studies suggest that ALC molted hens supported microflora and fermentation activities, which were more comparable to FF hens than FW hens by the end of the molting period.

[1]  V. Sheffield,et al.  Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Beier,et al.  Effect of dietary lactose on cecal pH, bacteriostatic volatile fatty acids, and Salmonella typhimurium colonization of broiler chicks. , 1990, Avian diseases.

[3]  S. Ricke,et al.  Limiting Avian Gastrointestinal Tract Salmonella Colonization by Cecal Anaerobic Bacteria, and a Potential Role for Methanogens , 2008 .

[4]  C. Wenk The role of dietary fibre in the digestive physiology of the pig , 2001 .

[5]  S. Ricke,et al.  Reduction of Salmonella enterica serovar enteritidis colonization and invasion by an alfalfa diet during molt in Leghorn hens. , 2005, Poultry science.

[6]  S. Ricke,et al.  The gastrointestinal tract ecology of Salmonella enteritidis colonization in molting hens. , 2003, Poultry science.

[7]  S. Ricke,et al.  Feed Deprivation Affects Crop Environment and Modulates Salmonella enteritidis Colonization and Invasion of Leghorn Hens , 1999, Applied and Environmental Microbiology.

[8]  J. Mattick,et al.  'Touchdown' PCR to circumvent spurious priming during gene amplification. , 1991, Nucleic acids research.

[9]  Y. Zo,et al.  Nonradioactive method to study genetic profiles of natural bacterial communities by PCR-single-strand-conformation polymorphism , 1996, Applied and environmental microbiology.

[10]  S. Ricke,et al.  Comparison of in vitro fermentation and molecular microbial profiles of high-fiber feed substrates incubated with chicken cecal inocula. , 2007, Poultry science.

[11]  A. E. Murray,et al.  Phylogenetic compositions of bacterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments , 1996, Applied and environmental microbiology.

[12]  S. Ricke,et al.  Effects of nitro compounds and feedstuffs on in vitro methane production in chicken cecal contents and rumen fluid. , 2006, Anaerobe.

[13]  K. Seo,et al.  Comparison of Salmonella Enteritidis infection in hens molted via long-term feed withdrawal versus full-fed wheat middling. , 2001, Journal of food protection.

[14]  B. Hargis,et al.  Provision of lactose to molting hens enhances resistance to Salmonella enteritidis colonization. , 1997, Journal of Food Protection.

[15]  S. Ricke,et al.  Identification and Quantification of Methanogenic Archaea in Adult Chicken Ceca , 2006, Applied and Environmental Microbiology.

[16]  G. Macfarlane,et al.  Gastrointestinal effects of prebiotics , 2002, British Journal of Nutrition.

[17]  N. Pace,et al.  Differential amplification of rRNA genes by polymerase chain reaction , 1992, Applied and environmental microbiology.

[18]  G. Muyzer,et al.  Genetic diversity of Desulfovibrio spp. in environmental samples analyzed by denaturing gradient gel electrophoresis of [NiFe] hydrogenase gene fragments , 1995, Applied and environmental microbiology.

[19]  C. S. DENATURING GRADIENT GEL ELECTROPHORESIS ( DGGE ) AS A RAPID METHOD FOR ASSESSING GASTROINTESTINAL TRACT MICROFLORA RESPONSES IN LAYING HENS FED SIMILAR ZINC MOLT INDUCTION DIETS , 2006 .

[20]  S. Ricke,et al.  Molting in Salmonella enteritidis-challenged laying hens fed alfalfa crumbles. I. Salmonella enteritidis colonization and virulence gene hilA response. , 2007, Poultry science.

[21]  A. Uitterlinden,et al.  Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA , 1993, Applied and environmental microbiology.

[22]  R. Joerger,et al.  16S rRNA-Based Analysis of Microbiota from the Cecum of Broiler Chickens , 2002, Applied and Environmental Microbiology.

[23]  J. Mcnab The Avian Caeca: A Review , 1973 .

[24]  S. Ricke,et al.  Comparison of zinc acetate and propionate addition on gastrointestinal tract fermentation and susceptibility of laying hens to Salmonella enteritidis during forced molt. , 2004, Poultry science.

[25]  K. E. Bach Knudsen,et al.  The nutritional significance of ''dietary fibre'' analysis , 2001 .

[26]  S. Ricke,et al.  Effect of various combinations of alfalfa and standard layer diet on susceptibility of laying hens to Salmonella enteritidis during forced molt. , 2006, Poultry science.

[27]  S. Pillai,et al.  Conventional and molecular methods for understanding probiotic bacteria functionality in gastrointestinal tracts. , 1999, Critical reviews in microbiology.

[28]  C. Donskey,et al.  Poultry digestive microflora biodiversity as indicated by denaturing gradient gel electrophoresis. , 2003, Poultry science.

[29]  S. Ricke,et al.  Evaluation of Salmonella enteritidis in molting hens after administration of an experimental chlorate product (for nine days) in the drinking water and feeding an alfalfa molt diet. , 2005, Poultry science.