Poult. Sci.
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Poult Sci 2008. 87:964-967. doi:10.3382/ps.2007-00287
© 2008 Poultry Science Association
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METABOLISM AND NUTRITION: Research Notes

The Effect of Gender on the Bacterial Community in the Gastrointestinal Tract of Broilers

B. S. Lumpkins, A. B. Batal1 and M. Lee

University of Georgia-Poultry Science, 208 Poultry Science Building 2772, Athens 30602-2772

1 Corresponding author: batal{at}uga.edu


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
The effect of gender on the growth, carcass yield, and nutritional requirements of chickens has been well documented, but little is known about how the sex of a chicken affects the bacterial population of the gastrointestinal tract. Therefore, our objective was to evaluate the biodiversity of the bacterial community in the gastrointestinal tract of male and female broilers. An experiment was conducted with Cobb 500 broiler chicks that were vent sexed at 0 d of age and allocated to 8 pens of 25 chicks per gender. All birds were fed a nonmedicated corn-soybean meal starter diet from 0 to 21 d of age. At 3, 7, 14, and 21 d of age, chicks were randomly selected and ileums were taken for bacterial sampling. Bacterial DNA was isolated from the digesta of the ileum, and denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of 16S ribosomal DNA. Denaturing gradient gel electrophoresis analyses revealed that the bacterial communities separated into 2 gender-specific groups, with less than 30% similarity between populations. Furthermore, as the birds aged, the similarity of the intestinal bacterial community decreased within each gender. Although ileal bacterial population differences within and between genders were noted as early as d 3, differences in growth rate between males and females were not noted until d 21 (data not shown). This suggested that non-growth-related factors influenced the composition of intestinal bacterial communities.

Key Words: bacterial community • gastrointestinal tract • denaturing gradient gel electrophoresis • gender


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
The gastrointestinal tract (GIT) is important for nutrient absorption and protection against many of the pathogens that enter the body (Mowat and Viney, 1997). The GIT also supports a microecosystem that harbors a large and diverse population of bacteria (Drasar and Barrow, 1985; Franks et al., 1998) that create a symbiotic relationship with the host (Apajalahti, 2005). The intraorganismal mutualism of the intestinal bacteria requires unique structures and environmental conditions of the GIT to become established and aid in the improvement of bird health (Cook and Bird, 1973; Fuller, 1989; Xu et al., 2003; Lan et al., 2005) and nutrient utilization for optimal growth (Lee et al., 1999). Therefore, not only is the structure of the GIT important for bird development, but so is the understanding of the symbiotic relationship between the bird and its intestinal bacterial community.

Many differences between male and female broilers have been reported, such as growth rate, deposition of muscle and bone minerals, and nutrient requirements (Han and Baker, 1993; Havenstein et al., 1994a,b; Rose et al., 1996; Horsted et al., 2005). Although there are no reports describing gender differences in GIT bacterial populations in poultry, Ley et al. (2005) observed that the amount of carcass fat influenced this characteristic in mice. In broilers, males have been reported to have leaner carcasses than females (Suto et al., 1998), which might be related to possible differences in the intestinal bacterial community. Considering that male and female broilers have different growth rates, one could hypothesize that the microecosystem of the GIT may also be different, which would influence the bacterial community. Therefore, the objective of this experiment was to evaluate the influence of gender on the bacterial population of broilers from 0 to 21 d of age.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS AND DISCUSSION
 REFERENCES
 
Bird Husbandry and Dietary Treatments
All procedures were approved by the University of Georgia Animal Use and Care Committee. Chicks were reared under standard commercial conditions and were provided with feed and water ad libitum. At hatch, broiler chicks (Cobb 500) were obtained from a local hatchery. Chicks were vent sexed and 25 of each gender were randomly allocated to 8 replicate (200 chicks per treatment) floor pens with litter shavings, with a dimension of 122 x 310 cm (density of 1,511 cm2 per bird). The birds were housed in an environmentally controlled room under 24-h lighting conditions with Ziggity nipple drinkers (Middlebury, IN) and Chore-Time feeders (Milford, IN). Chicks were fed a nonmedicated conventional corn-soybean meal starter diet (23% CP, 3,096 kcal of ME/kg, 1.14% total lysine, 0.53% total methionine, and 0.90% TSAA) from 0 to 21 d of age.

Bacterial DNA Isolation
Two birds per pen were euthanized and ileum (from the yolk stalk to the ileocecal junction) samples were taken at 3, 7, 14, and 21 d of age. The bacterial portion of the ileal contents was recovered by density gradient centrifugation through a gauze column, and the DNA was extracted as described by Lu et al. (2003).

Denaturing Gradient Gel Electrophoresis
The V3 to V4 region of the 16S ribosomal DNA of microorganisms of the bacterial domain from contents of the chicken ileum were amplified with the primers HDA1-GC and HDA2 as described by Knarreborg et al. (2002). Amplicons were confirmed by visualization of electrophoresis by using a 1.5% agarose gel containing ethidium bromide. Deoxyribonucleic acid sequence polymorphisms of the amplicons were detected by resolving differences in molecular structure by using denaturing gradient gel electrophoresis (DGGE). The DGGE was conducted by using the D-Code Universal Mutation Detection system (Bio-Rad, Hercules, CA) with 16 cm x 16 cm x 1 mm gels composed of 8% (wt/vol) polyacrylamide (acrylamide:bis, 37.5:1) gels in 1x Tris-acetate-EDTA buffer with a 15 to 55% linear denaturant gradient. The 100% denaturing solution contained 40% (vol/vol) formamide and 7.0 M urea. Electrophoresis was performed at a constant voltage of 200 V at 60°C for 3 h. Gels were fixed in 10% acetic acid for 15 min, washed 3 times in deionized water, then put in 50% methanol, washed, and stained in 5 µg of Sybr Green I (FMC Bio Products, Philadelphia, PA) per milliliter of 1x Tris-acetate-EDTA buffer for 30 min. After staining, the gels were analyzed by using a laser densitometer FluorImage (Molecular Dynamics, Sunnyvale, CA) with Fragment Analysis software (Capture One Pro Imaging Software, Phase One, Melville, NY).

Estimates of Diversity
After separation of amplicons by DGGE, their profiles were compared by using a coefficient of similarity (Cs), which was determined as Cs = [2j/(a + b)] x 100 (McCracken et al., 2001), where a is the number of bands in a sample (i.e., lane on the gel), b is the number of bands in another sample, and j is the number of common DGGE bands. Therefore, if the banding profiles of the 2 lanes are identical, Cs = 100%, whereas Cs = 0% for 2 samples (lanes) with completely different banding profiles. After determination of the various Cs values, a dendrogram (evolutionary tree) was constructed by using Treecon version 1.3b for Windows (Van de Peer and De Wachter, 1994, 1997).


   RESULTS AND DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
REFERENCES
 
Body weight gain (BWG) was not significantly different (P > 0.05) from 0 to 14 d of age between sexes (data not shown). However, by 21 d of age, males exhibited greater (P < 0.05) growth rates than females. Considering that there were no differences in intestinal development or BWG between genders before this age, it might be concluded that their intestinal environments, and associated bacterial communities, might be similar. However, on the basis of the bacterial analysis, there was less than a 30% similarity in the bacterial populations of males and females (Figure 1Go).


Figure 1
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  		Figure 1. Dendrogram representing the similarity among ileal bacterial communities in male and female broilers from 3 to 21 d of age. Simple matching was performed by using the clustering method of Treecon to produce this dendrogram, which depicts the relationship between denaturing gradient gel electrophoresis band patterns of the V3 to V4 region of 16S ribosomal DNA of the intestinal bacterial communities of male and female broilers of different ages (Van de Peer and De Wachter, 1994, 1997). If 2 band profiles were identical, then the bacterial communities had a similarity of 100%. Conversely, samples with completely different banding patterns had a similarity of 0%. For each gender-age combination, the ileal contents of 16 birds were pooled and subjected to bacterial analysis.

 
The reason for the difference in the bacterial communities between genders is not well understood and may involve a number of microenvironmental conditions such as pH, temperature, and mucin composition or concentration that would provide various niches and influence the type of bacteria that can successfully compete for nutrients in the intestine. There are few published reports on the composition of the intestinal bacteria of different genders of animals, and none on poultry. Our findings support the previous observation of Ge et al. (2006), who reported that the bacterial communities of male and female Swiss Webster mice differed. Specifically, as compared with males, female mice had a different species of Lactobacillus present in their bacterial community, which was reported to be a contributing factor to the female’s ability to respond immunologically to Helicobacter hepaticus infection. Therefore, differences in disease resistance between the sexes could be partly due to differences in the bacterial population present in the GIT. Furthermore certain species of bacteria present within the intestinal bacterial community could affect the health of the animal by improving its response to a specific infection, as suggested by Ge and colleagues, acting in intraorganismal mutualism (Lan et al., 2005).

In addition to the impact of gender, there was a definite age effect on the bacterial community (Figure 1Go). In males, the bacterial community was 70% similar between 3 and 7 d of age and between 14 and 21 d of age, but there was a decrease in similarity to 45% when the bacterial community was compared at either 3 or 7 d of age with older birds at either 14 or 21 d of age. The bacterial community in the females followed a more stepwise trend, with the bacterial community becoming less similar as the bird aged, beginning at 90% similarity between 3 and 7 d of age and moving to 50% similarity between 3 and 21 d of age. Lu et al. (2003) and Amit-Romach et al. (2004) also reported a change in the composition of the bacterial community as the bird ages; however, the gender of the birds was not taken into consideration.

In conclusion, the biodiversity of the microbiota in both genders changed as the birds aged. The similarity in BWG and intestinal measurements between genders during the early stages of growth (data not shown) would suggest similar intestinal microecosystems or bacterial communities. However, this was not borne out by the DDGE analyses because the intestinal bacterial populations clearly differed according to gender. Moreover, our work suggests that in chickens, both within and between genders, factors unrelated to body growth rate, intestinal development, or both are responsible for differences in intestinal bacterial communities. Future research will be necessary to gain a better understanding of what influences bacterial populations in the GIT.

Received for publication July 16, 2007. Accepted for publication February 2, 2008.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
Amit-Romach, E., D. Sklan, and Z. Uni. 2004. Microflora ecology of the chicken intestine using 16S ribosomal DNA primers. Poult. Sci. 83:1093–1098.[Abstract/Free Full Text]

Apajalahti, J. 2005. Comparative gut microflora, metabolic challenges, and potential opportunities. J. Appl. Poult. Res. 14:444–453.[Abstract/Free Full Text]

Cook, R. H., and F. H. Bird. 1973. Duodenal villus area and epithelial cellular migration in conventional and germfree chicks. Poult. Sci. 52:2276–2280.[Web of Science][Medline]

Drasar, B. S., and P. A. Barrow. 1985. Intestinal Microbiology. Am. Soc. Microbiol., Washington, DC.

Franks, A. H., H. J. M. Harmsen, G. C. Rangs, G. J. Jansen, F. Schut, and G. W. Welling. 1998. Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rRNA-targeted oligo-nucleotide probes. Appl. Environ. Microbiol. 64:3336–3345.[Abstract/Free Full Text]

Fuller, R. 1989. A review: Probiotics in man and animal. J. Appl. Bacteriol. 66:365–378.[Medline]

Ge, Z. M., Y. Feng, N. S. Taylor, M. Ohtani, M. F. Polz, D. B. Schauer, and J. G. Fox. 2006. Colonization dynamics of altered Schaedler flora is influenced by gender, aging, and Helicobacter hepaticus infection in the intestines of Swiss Webster mice. Appl. Environ. Microbiol. 72:5100–5103.[Abstract/Free Full Text]

Han, Y., and D. H. Baker. 1993. Effects of sex, heat stress, body weight, and genetic strain on the lysine requirement of broiler chicks. Poult. Sci. 72:701–708.[Web of Science][Medline]

Havenstein, G. B., P. R. Ferket, S. E. Scheideler, and B. T. Larson. 1994a. Carcass composition and yield of 1991 vs. 1957 broilers when fed typical 1957 and 1991 broiler diets. Poult. Sci. 73:1795–1804.[Web of Science][Medline]

Havenstein, G. B., P. R. Ferket, S. E. Scheideler, and B. T. Larson. 1994b. Growth, livability, and feed conversion of 1957 vs. 1991 broilers when fed typical 1957 and 1991 broiler diets. Poult. Sci. 73:1785–1794.[Web of Science][Medline]

Horsted, K., J. Henning, and J. E. Hermansen. 2005. Growth and sensory characteristics or organically reared broilers differing in strain, sex and age at slaughter. Acta Agric. Scand. Anim. Sci. 55:149–157.[CrossRef]

Knarreborg, A., M. A. Simon, R. M. Engberg, B. B. Jensen, and G. W. Tannock. 2002. Effects of dietary fat source and subtherapeutic levels of antibiotic on the bacterial community in the ileum of broiler chickens at various ages. Appl. Environ. Microbiol. 68:5918–5924.[Abstract/Free Full Text]

Lan, Y., M. W. A. Verstegen, S. Tamminga, and B. A. Williams. 2005. The role of the commensal gut microbial community in broiler chickens. World’s Poult. Sci. J. 61:95–104.[CrossRef][Web of Science]

Lee, Y. K., K. Nomoto, S. Salminen, and S. L. Gorbach. 1999. Handbook of Probiotics. John Wiley and Sons Inc., New York, NY.

Ley, R. E., F. Backhead, P. Turnbaugh, C. A. Lozupone, R. D. Knight, and J. I. Gordon. 2005. Obesity alters gut microbial ecology. Proc. Natl. Acad. Sci. USA 102:11070–11075.[Abstract/Free Full Text]

Lu, J., U. Idris, D. Harmon, C. Hofacre, J. J. Maurer, and M. D. Lee. 2003. Diversity and succession of the intestinal bacterial community of maturing broiler chicken. Appl. Environ. Microbiol. 69:6816–6824.[Abstract/Free Full Text]

McCracken, V. J., J. M. Simpson, R. I. Mackie, and H. R. Gaskins. 2001. Molecular ecological analysis of dietary and antibiotic-induced alterations of the mouse intestinal microbiota. J. Nutr. 131:1862–1870.[Abstract/Free Full Text]

Mowat, A. M., and J. L. Viney. 1997. The anatomical basis of intestinal immunity. Immunol. Rev. 156:145–166.[CrossRef][Web of Science][Medline]

Rose, N., P. Constantin, and C. Leterrier. 1996. Sex difference in bone growth of broiler chickens. Growth Dev. Aging 60:49–59.[Web of Science][Medline]

Suto, Z., P. Horn, J. F. Jensen, P. Sorensen, and J. Csapo. 1998. Carcass traits, abdominal fat deposition and chemical composition of commercial meat type chicken during a twenty week growing period. Arch. Geflugelkd. 62:21–25.

Van de Peer, Y., and R. De Wachter. 1994. TREECON for Windows: A software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput. Applic. Biosci. 10:569–570.[Free Full Text]

Van de Peer, Y., and R. De Wachter. 1997. Construction of evolutionary distance trees with TREECON for Windows: Accounting for variation in nucleotide substitution rate among sites. Comput. Applic. Biosci. 13:227–230.[Abstract/Free Full Text]

Xu, Z. R., C. H. Hu, M. S. Xia, X. A. Zhan, and M. Q. Wang. 2003. Effects of dietary fructooligosaccharide on digestive enzymes and morphology of male broilers. Poult. Sci. 82:1030–1036.[Abstract/Free Full Text]





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