Animal Science Publications Animal Science 1-1-2008 Association of Interleukin-10 Cluster Genes and Salmonella Response in the Chicken S. B. Ghebremicael Iowa State University J. Hasenstein Iowa State University Susan J. Lamont Iowa State University, [email protected] Follow this and additional works at: https://lib.dr.iastate.edu/ans_pubs Part of the Agriculture Commons, Genetics Commons, and the Poultry or Avian Science Commons The complete bibliographic information for this item can be found at https://lib.dr.iastate.edu/ ans_pubs/607. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Article is brought to you for free and open access by the Animal Science at Iowa State University Digital Repository. It has been accepted for inclusion in Animal Science Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Association of Interleukin-10 Cluster Genes and Salmonella Response in the Chicken Abstract Salmonella enteritidis lipopolysaccharide stimulates interleukin 10 (IL10) gene expression in chickens. Four genes in the IL10 cluster [polymeric immunoglobulin receptor (PIGR), interleukin 10 (IL10), map kinase-activated protein kinase 2 (MAPKAPK2), and ligatin (LGTN)] plus dual-specificity tyrosine-(Y)- phosphorylation regulated kinase1A (DYRK1A) were investigated using the F8 generation of 2 related advanced intercross lines (AIL). The AIL were generated by crossing outbred broilers with dams of 2 highly inbred lines (Leghorn and Fayoumi). Intercrossing continued within the 2 dam lines. The F8 chicks (n = 132) were intraesophageally inoculated at 1 d with S. enteritidis. At d 7 or 8, both spleen tissue and cecal contents were cultured to quantify S. enteritidis load. The F8 population was genotyped for one single nucleotide polymorphism (SNP) per gene by using a multiplexed SNaPshot assay. Association of gene SNP with S. enteritidis bacterial burden was analyzed by the GLM. The MAPKAPK2 and IL10 genes were highly (P < 0.001) associated with S. enteritidis burden in spleen tissue and cecal luminal content. Suggestive associations (P < 0.05) of PIGR with spleen tissue and cecal content were found. The results suggest that SNP in MAPKAPK2 and IL10 were strongly associated with Salmonella burden and may be valuable in generating resistant birds by marker-assisted selection. Keywords lipopolysaccharide, interleukin-10 cluster genes, Salmonella, advanced intercross line, single nucleotide polymorphism Disciplines Agriculture | Animal Sciences | Genetics | Poultry or Avian Science Comments This article is published as Ghebremicael, S. B., J. R. Hasenstein, and S. J. Lamont. "Association of interleukin-10 cluster genes and Salmonella response in the chicken." Poultry Science 87, no. 1 (2008): 22-26. DOI: 10.3382/ps.2007-00259. Posted with permission. This article is available at Iowa State University Digital Repository: https://lib.dr.iastate.edu/ans_pubs/607 GENETICS Research Note Association of Interleukin-10 Cluster Genes and Salmonella Response in the Chicken S. B. Ghebremicael, J. R. Hasenstein, and S. J. Lamont Department of Animal Science, Iowa State University, Ames 50011 ABSTRACT Salmonella enteritidis lipopolysaccharide and cecal contents were cultured to quantify S. enteritidis stimulates interleukin 10 (IL10) gene expression in chick- load. The F8 population was genotyped for one single ens. Four genes in the IL10 cluster [polymeric immuno- nucleotide polymorphism (SNP) per gene by using a mul- globulin receptor (PIGR), interleukin 10 (IL10), map ki- tiplexed SNaPshot assay. Association of gene SNP with nase-activated protein kinase 2 (MAPKAPK2), and ligatin S. enteritidis bacterial burden was analyzed by the GLM. (LGTN)] plus dual-specificity tyrosine-(Y)-phosphoryla- The MAPKAPK2 and IL10 genes were highly (P < 0.001) tion regulated kinase1A (DYRK1A) were investigated us- associated with S. enteritidis burden in spleen tissue and < ing the F8 generation of 2 related advanced intercross cecal luminal content. Suggestive associations (P 0.05) lines (AIL). The AIL were generated by crossing outbred of PIGR with spleen tissue and cecal content were found. broilers with dams of 2 highly inbred lines (Leghorn and The results suggest that SNP in MAPKAPK2 and IL10 Fayoumi). Intercrossing continued within the 2 dam lines. were strongly associated with Salmonella burden and may The F8 chicks (n = 132) were intraesophageally inoculated be valuable in generating resistant birds by marker-as- at 1 d with S. enteritidis. At d 7 or 8, both spleen tissue sisted selection. Key words: lipopolysaccharide, interleukin-10 cluster genes, Salmonella, advanced intercross line, single nucleotide polymorphism 2008 Poultry Science 87:22–26 doi:10.3382/ps.2007-00259 INTRODUCTION competitive exclusion and vaccination. The use of antibiot- ics in domestic livestock has been questioned due to the Salmonella contamination of poultry meat and eggs con- possible creation of antibiotic-resistant bacteria and the tinues to be a global threat to public health (Barrow and possibility of antibiotic residues being consumed by hu- Duchet-Suchaux, 1997). Salmonella enteritidis is an enteric mans (White et al., 2001). bacterium that is a zoonotic intracellular pathogen of poul- Host genetic factors clearly influence the epidemiology try and humans as well as other species (McIlroy et al., of S. enteritidis infection in chickens. Newer strategies to 1989). Salmonella enteritidis is a major cause of human increase the overall level of resistance at flock and popula- foodborne illness and is the most frequent serovar detected tion levels that use selective breeding programs to enhance in outbreaks of human salmonellosis (Lu et al., 1999). In natural resistance are expected to contribute significantly the United States, an estimated 500,000 cases of human in this regard (van der Zjipp, 1983; Warner et al., 1987; illness are annually attributed to S. enteritidis due to con- Womack, 1988). In recent years, much progress on the taminated food products (Schlosser and Ebel, 1998). In the identification and characterization of candidate genes, mi- advent of S. enteritidis infection, some animals die, but crosatellite markers, and comparative gene mapping has others can host the bacteria for several weeks or months been made. Identification of individual candidate genes without presenting any particular symptoms. These that control natural resistance and the actions of these healthy carriers are therefore an obstacle to the eradication genes will greatly expand our knowledge of genetic resis- tance to bacterial diseases and the possibilities for practical of S. enteritidis and are responsible for the transmission application. A candidate gene is usually chosen, based to humans. Different prophylactic measures to control S. on its biological or physiological functions, to study the enteritidis infection in poultry have been studied, including association between its genetic polymorphisms and traits of interest (Rothschild and Soller, 1997). Once a candidate gene is identified and proven, causative mutations (func- © 2008 Poultry Science Association Inc. tional markers) or polymorphic markers (direct markers) Received June 21, 2007. Accepted September 27, 2007. near or within the gene sequence can be developed for 1Corresponding author: [email protected] marker-assisted selection to improve immunity. 22 RESEARCH NOTE 23 The ability to respond to S. enteritidis infections is age- lines (Leghorn and Fayoumi; Zhou and Lamont, 1999). dependent in poultry. In young chickens, salmonellosis is Further intercrossing of F1 birds within the 2 granddam a major disease characterized by severe clinical signs of lines yielded the F8 birds. The genetic distance between diarrhea and dehydration with high mortality rates. In the parental lines maximized the feasibility of finding mo- adult chickens, it does not cause significant disease or lecular genetic polymorphism, and the inbred nature of mortality, and birds can carry the bacteria for several the founder dams meant that they consistently contributed weeks without presenting any clinical signs, which consti- the same allele to all offspring. To ensure that the maternal tutes a risk for public health (Wigley et al., 2002). The immune status of all hens producing the chicks was equiv- heritability estimates of S. enteritidis range from 0.13 to alent and would not interfere with testing their chicks for 0.53 for frequency in internal organs of challenged adult Salmonella response, all hens were kept under the same layers (Beaumont et al., 1999), 0.20 in chickens inoculated biosecure management conditions. Chicks were tested for at 1 wk of age, 0.38 in hens inoculated at the peak of lay Salmonella before challenge. The chicks were equally di- (Berthelot et al., 1998; Beaumont et al., 1999), and 0.10 for vided with regard to genetic line and sex into 3 biosafety- spleen infection using 13-d-old chicks (Girard-Santosuosso level-2 animal rooms. et al., 2002). These estimated heritabilities suggest that re- sistant populations could be developed through selection Salmonella Pathogenic Challenge for a lower level of contamination of spleen and cecum. and Quantification of Bacterial Load Over the past 10 yr numerous candidate gene studies have found allelic variation to affect the immune response to S. The F8 chicks (n = 132) were intraeseophageally inocu- enteritidis in poultry (Hu et al., 1997; Cotter