Avian Beta-defensin Research Articles

The effect of humic substances on gut microbiota and immune response of broilers

ABSTRACT The work focused on the study of the immunomodulatory and gut-protecting effect of humic substances (HS) in broilers. The diet of experimental chicks was enriched with 0.8% of HS. We noted that HS had a stimulatory effect on the phagocytic activity and the engulfing capacity of phagocytes, however, the level of oxidative burst of phagocytes was not affected. We observed a significant increase of CD4+: CD8+ lymphocyte ratio, an indicator of immune stimulation. HS did not influence the IgA gene expression. In contrast, we observed a significant increase in the expression of MUC-2 (intestinal mucin 2) gene, and a decrease in the expression of IGF-2 (insulin-like growth factor 2) and also AvBD2 (avian beta defensin 2) genes. A decreased Enterobacteriaceae counts in the gut of experimental animals showed a positive effect on intestinal microbiota. We confirmed a gut-protecting and an immunostimulatory effect of HS in broiler chickens.

Systemic Administration of Avian Defensin 7: Distribution, Cellular Target, and Antibacterial Potential in Mice.

Defensins are natural antimicrobial peptides. The avian beta-defensin AvBD7 isolated from the chicken bone marrow possess broad antibacterial spectrum and strong resistance to proteolysis. However, its ability to fight systemic infections of major concern for public health, such as salmonellosis, is unknown. As a first approach, fluorescence labeling of AvBD7 allowed to track its systemic distribution after intraperitoneal injection in mice using whole body live imaging. It was associated to peritoneal cells and to deeper organs such as the liver. In the next step, the use of labeled AvBD7 allowed to observe its interaction with murine macrophages in culture. After incubation, it was able to penetrate inside the cells through an endocytosis-like mechanism. Furthermore, natural AvBD7 contributed to the control of intracellular multiplication of a multidrug resistant Salmonella strain, after incubation with infected macrophages. Finally, administration in a model of systemic lethal Salmonella infection in mice led to significant improvement of mouse survival, consistently with significant reduction of the liver bacterial load. In conclusion, the results reveal a hitherto unknown intracellular antibacterial effect of AvBD7 in Salmonella target cells and support AvBD7 as a candidate of interest for the treatment of infectious diseases caused by multidrug-resistant pathogenic Enterobacteriaceae.

The Unique Features of Proteins Depicting the Chicken Amniotic Fluid

In many amniotes, the amniotic fluid is depicted as a dynamic milieu that participates in the protection of the embryo (cushioning, hydration, and immunity). However, in birds, the protein profile of the amniotic fluid remains unexplored, even though its proteomic signature is predicted to differ compared with that of humans. In fact, unlike humans, chicken amniotic fluid does not collect excretory products and its protein composition strikingly changes at mid-development because of the massive inflow of egg white proteins, which are thereafter swallowed by the embryo to support its growth. Using GeLC-MS/MS and shotgun strategies, we identified 91 nonredundant proteins delineating the chicken amniotic fluid proteome at day 11 of development, before egg white transfer. These proteins were essentially associated with the metabolism of nutrients, immune response and developmental processes. Forty-eight proteins were common to both chicken and human amniotic fluids, including serum albumin, apolipoprotein A1 and alpha-fetoprotein. We further investigated the effective role of chicken amniotic fluid in innate defense and revealed that it exhibits significant antibacterial activity at day 11 of development. This antibacterial potential is drastically enhanced after egg white transfer, presumably due to lysozyme, avian beta-defensin 11, vitelline membrane outer layer protein 1, and beta-microseminoprotein-like as the most likely antibacterial candidates. Interestingly, several proteins recovered in the chicken amniotic fluid prior and after egg white transfer are uniquely found in birds (ovalbumin and related proteins X and Y, avian beta-defensin 11) or oviparous species (vitellogenins 1 and 2, riboflavin-binding protein). This study provides an integrative overview of the chicken amniotic fluid proteome and opens stimulating perspectives in deciphering the role of avian egg-specific proteins in embryonic development, including innate immunity. These proteins may constitute valuable biomarkers for poultry production to detect hazardous situations (stress, infection, etc.), that may negatively affect the development of the chicken embryo.

High dietary energy content increases inflammatory markers after lipopolysaccharide challenge in meat ducks

This study was conducted to investigate the effect of dietary energy concentration on growth performance, serum biochemical indices, and the mRNA expression of inflammatory cytokines in the liver of meat ducks challenged with lipopolysaccharide (LPS). A total of 600 1-d-old Cherry Valley meat ducks were randomly assigned to 10 treatments with six replicate cages of 10 birds/pen. A 5 × 2 factorial arrangement in a complete randomized design experiment was used to study the effect of five concentrations of dietary metabolizable energy (ME, 2,800, 2,900, 3,000, 3,100, and 3,200kcal/kg) with or without a challenge with LPS in meat ducks. All experimental ducks were intraperitoneally injected with either 0.5mg/kg body weight (BW) LPS or an equivalent amount of sterile saline at 15, 17, and 19 d of age. The results showed that LPS challenge significantly decreased (P < 0.05) BW (d 21), body weight gain (d 15-21) and average daily feed intake (ADFI; d 15-21), and markedly increased (P < 0.05) relative spleen weight, serum total protein concentration, and the mRNA expression of interleukin 6 (IL-6), interleukin 10, transforming growth factor β (TGF-β), and Avian Beta Defensin 10 (AVBD-10) in the liver of meat ducks. Ducks fed 3,200kcal/kg ME diet had the lowest (P < 0.05) ADFI than those fed with the other diets. Additionally, there were significantly LPS by diet interactions on IL-6, interferon-γ, TGF-β, AVBD-10, and inducible nitric oxide synthase mRNA expression (P < 0.05), wherein a more pronounced liver inflammatory response was observed in birds fed high-energy diets (3,100 and 3,200kcal/kg ME) than birds fed low-energy diet (2,800 and 2,900kcal/kg ME). The results indicated that 3,100 and 3,200kcal/kg ME of diets increased hepatic inflammatory markers induced by LPS challenge in ducks but did not influence performance responsiveness during this challenge.

Differential expression of intestinal nutrient transporters and host defense peptides in Eimeria maxima-infected Fayoumi and Ross chickens

Fayoumi chickens are believed to be more disease resistant compared to commercial broiler chickens. The objective of this study was to compare mRNA expression of intestinal nutrient transporters, digestive enzymes, and host defense peptides (HDP) between Eimeria maxima-challenged Fayoumi and Ross broiler chickens. At 21 d of age, Ross broilers and Fayoumi lines M5.1 and M15.2 were challenged with 1,000 E. maxima oocysts. Control birds were not challenged. Duodenum, jejunum, and ileum were sampled (n = 6) at 7 d post challenge. Gene expression was analyzed using relative quantification PCR. Data were analyzed by ANOVA and significance level was set at P < 0.05. There was numerical, but not statistically significant, differential weight gain depression for Ross (15%) and Fayoumi lines M5.1 (21%) and M15.2 (22%) and significant line-specific changes in gene expression. For nutrient transporters, there was downregulation of mRNA for the brush border membrane, amino acid transporters b0,+AT/rBAT, BoAT, and EAAT3 in different segments of the small intestine of Ross and both lines of Fayoumi chickens, indicating that E. maxima challenge likely caused a decrease in nutrient uptake. For HDP, there was downregulation of avian beta defensin (AvBD) 1, 6, 10, 12, and 13 mRNA in the jejunum of the 2 Fayoumi lines, but no change in the Ross broilers. In the duodenum, there was upregulation of AvBD10 mRNA in the Ross and both Fayoumi lines and additionally upregulation of AvBD11, 12, and 13 mRNA in only Fayoumi line M15.2. Liver expressed antimicrobial peptide 2 (LEAP2) mRNA was downregulated in the duodenum and jejunum of Ross and Fayoumi line M5.1 but not in Fayoumi line M15.2. The homeostatic, non-challenged levels of AvBD mRNA were greater in Fayoumi line M15.2 than Ross and Fayoumi line M5.1 in the duodenum and ileum. This study demonstrates tissue- and genetic line-specific transcriptional responses to E. maxima, highlights novel potential candidate genes for response to coccidiosis, and confirms a role for several previously reported genes in response to coccidiosis.

The role of the outer membrane protein gene ybjX in the pathogenicity of avian pathogenic Escherichia coli

ABSTRACT Avian pathogenic Escherichia coli (APEC) are important pathogens that cause serious economic losses to the poultry industry worldwide. The PhoP/Q system is essential for APEC pathogenicity, and its mutation leads to a decrease in ybjX gene expression, but whether ybjX regulated by PhoP/Q plays a role in pathogenicity remains unclear. The role of ybjX in APEC pathogenicity was investigated using a ybjX mutant strain constructed from APEC strain AE17 by the Red recombination method. The mutant strain displayed significantly reduced adherence to and invasion of chicken embryo fibroblast cell line (DF-1) cells. Moreover, N-phenyl-1-naphthylamine (NPN) assays revealed an increase in outer membrane permeabilization for the ybjX mutant, suggesting ybjX affected susceptibility to avian beta-defensin through this mechanism. The ybjX mutation increased susceptibility to avian beta-defensin 9 (AvBD9), and in vivo LD50 assays revealed reduced bacterial virulence in a chicken model. Taken together, these results indicated that mutation of ybjX may attenuate APEC pathogenicity, not only by affecting bacterial adhesion and invasion, but also via reducing resistance to avian beta-defensin by increasing outer membrane permeabilization of bacteria.

Expression and regulation of avian beta-defensin 8 protein in immune tissues and cell lines of chickens.

ObjectiveDefensins are a large family of antimicrobial peptides and components of the innate immune system that invoke an immediate immune response against harmful pathogens. Defensins are classified into alpha-, beta-, and theta-defensins. Avian species only possess beta-defensins (AvBDs), and approximately 14 AvBDs (AvBD1–AvBD14) have been identified in chickens to date. Although substantial information is available on the conservation and phylogenetics, limited information is available on the expression and regulation of AvBD8 in chicken immune tissues and cells.MethodsWe examined AvBD8 protein expression in immune tissues of White Leghorn chickens (WL) by immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (RT-qPCR). In addition, we examined AvBD8 expression in chicken T-, B-, macrophage-, and fibroblast-cell lines and its regulation in these cells after lipopolysaccharide (LPS) treatment by immunocytochemistry and RT-qPCR.ResultsOur results showed that chicken AvBD8 protein was strongly expressed in the WL intestine and in macrophages. AvBD8 gene expression was highly upregulated in macrophages treated with different LPS concentrations compared with that in T- and B-cell lines in a time-independent manner. Moreover, chicken AvBD8 strongly interacted with other AvBDs and with other antimicrobial peptides as determined by bioinformatics.ConclusionOur study provides the expression and regulation of chicken AvBD8 protein in immune tissues and cells, which play crucial role in the innate immunity.

Expression of host defense peptides in the intestine of Eimeria-challenged chickens

Avian coccidiosis is caused by the intracellular protozoan Eimeria, which produces intestinal lesions leading to weight gain depression. Current control methods include vaccination and anticoccidial drugs. An alternative approach involves modulating the immune system. The objective of this study was to profile the expression of host defense peptides such as avian beta-defensins (AvBDs) and liver expressed antimicrobial peptide 2 (LEAP2), which are part of the innate immune system. The mRNA expression of AvBD family members 1, 6, 8, 10, 11, 12, and 13 and LEAP2 was examined in chickens challenged with either E. acervulina, E. maxima, or E. tenella. The duodenum, jejunum, ileum, and ceca were collected 7 d post challenge. In study 1, E. acervulina challenge resulted in down-regulation of AvBD1, AvBD6, AvBD10, AvBD11, AvBD12, and AvBD13 in the duodenum. E. maxima challenge caused down-regulation of AvBD6, AvBD10, and AvBD11 in the duodenum, down-regulation of AvBD10 in the jejunum, but up-regulation of AvBD8 and AvBD13 in the ceca. E. tenella challenge showed no change in AvBD expression in any tissue. In study 2, which involved challenge with only E. maxima, there was down-regulation of AvBD1 in the ileum, AvBD11 in the jejunum and ileum, and LEAP2 in all 3 segments of the small intestine. The expression of LEAP2 was further examined by in situ hybridization in the jejunum of chickens from study 2. LEAP2 mRNA was expressed similarly in the enterocytes lining the villi, but not in the crypts of control and Eimeria challenged chickens. The lengths of the villi in the Eimeria challenged chickens were less than those in the control chickens, which may in part account for the observed down-regulation of LEAP2 mRNA quantified by PCR. Overall, the AvBD response to Eimeria challenge was not consistent; whereas LEAP2 was consistently down-regulated, which suggests that LEAP2 plays an important role in modulating an Eimeria infection.

Comparative expression of host defense peptides in turkey poults

Host defense peptides (HDPs) are a large group of small, positively charged peptides that play an important role in innate immunity, particularly at early ages when other components of the immune system have not fully developed. There are 3 classes of avian HDPs: avian beta defensins (AvBDs), cathelicidins (Cath), and liver-expressed antimicrobial peptide 2 (LEAP-2). The objective was to compare expression of HDP mRNAs in male turkey poults at day of hatch (d 0), d 7, d 14, d 21 and d 28 from the thymus, spleen, bursa, duodenum, jejunum, and ileum. The expression of AvBD1, AvBD2, AvBD8, AvBD9, AvBD10, AvBD13, Cath2, Cath3, and LEAP-2 mRNA was measured using qPCR (n = 6 birds/tissue/age). Data were analyzed by one-way ANOVA and Tukey’s test, and significance considered at P < 0.05. AvBDs and Caths exhibited greater expression in immune organs (thymus, spleen, and bursa) than intestinal tissues. In the thymus, expression of all AvBDs examined, except AvBD8, showed an increase from d 0 to d 21. In the spleen, AvBD1 and AvBD2 exhibited reduced expression from d 0 to d 7 and low expression thereafter. In the intestine, AVBD1, AVBD8, and AvBD13 increased expression from d 0 to d 28 in the duodenum, while AvBD10 showed the greatest expression at d 0 that declined to d 7 and stayed low thereafter in the duodenum, jejunum, and ileum. Cath2 and Cath3 demonstrated the highest expression in the spleen, which was greatest at d 0 then declined to d 7 through d 28. Conversely, LEAP-2 showed greater expression in the intestinal tissues than in the immune organs. LEAP-2 expression was upregulated from d 0 to d 7 and then remained elevated from d 7 through d 14 in the duodenum. In the jejunum, LEAP-2 increased from d 0 to d 21 and d 28. Understanding the differential expression of HDPs could reveal the innate immune status of turkey poults, and may subsequently allow improvement of their health through appropriate mitigation strategies.

Haplotype structure and copy number polymorphism of the beta-defensin 7 genes in diverse chicken breeds.

Beta-defensins is a family of avian peptides related to the innate immune system. Copy number variation was recently reported for the avian beta-defensin 7 gene (AvBD7) between the highly inbred Leghorn and Fayoumi lines. Here, we examined copy number variants in 35 different chicken breeds and found that 31 of them have at least the same representation of the duplicated AvBD7 allele. We also found haplotypes upstream of the AvBD6 regions that are strongly linked to the AvBD7 duplication. We observed a strong linkage disequilibrium spanning of the upstream region of the AvBD6 gene, with two SNPs being flanking markers to detect duplication of the AvBD7.

Immune responses to in ovo vaccine formulations containing inactivated fowl adenovirus 8b with poly[di(sodium carboxylatoethylphenoxy)]phosphazene (PCEP) and avian beta defensin as adjuvants in chickens

Inclusion body hepatitis (IBH) is one of the major viral infections causing substantial economic loss to the global poultry industry. The disease is characterized by a sudden onset of mortality (2–30%) and high morbidity (60–70%). IBH is caused by a number of serotypes of fowl adenovirus with substantially low levels of serotype cross protection. Thus far, there is no effective and safe vaccine commercially available in the North America for the control of IBH in chickens.Poly[di(sodium carboxylatoethylphenoxy)]phosphazene (PCEP) is a high molecular weight, biodegradable water soluble polymer that has been well characterized as a safe and effective adjuvant for a number of experimental veterinary vaccines. Similarly, host defence peptides, including β-defensins, have also been shown to exhibit strong adjuvant potential.In this study, we evaluated the adjuvant activity of PCEP and avian beta defensin (ABD) in a vaccine formulation containing inactivated fowl adenovirus (FAdV) serotype 8b administered in ovo.Our data showed that a combination of PCEP and inactivated virus is capable of inducing a robust and long lasting antibody response. Moreover, significant enhancement of IFN-γ, IFN-α, IL-12(p40) and IL-6 gene expression under the influence of PCEP suggests that as an in ovo adjuvant PCEP has the ability to activate a substantial balanced immune response in chickens. To our knowledge, these are the first studies in which PCEP and ABD have been characterized as adjuvants for the development of an in ovo poultry vaccine. It is expected that these preliminary studies will be helpful in the development of safer and more effective in ovo vaccine against IBH and other infectious diseases affecting chickens.

Structure-function analysis of Avian β-defensin-6 and β-defensin-12: role of charge and disulfide bridges.

BackgroundAvian beta-defensins (AvBD) are small, cationic, antimicrobial peptides. The potential application of AvBDs as alternatives to antibiotics has been the subject of interest. However, the mechanisms of action remain to be fully understood. The present study characterized the structure-function relationship of AvBD-6 and AvBD-12, two peptides with different net positive charges, similar hydrophobicity and distinct tissue expression profiles.ResultsAvBD-6 was more potent than AvBD-12 against E. coli, S. Typhimurium, and S. aureus as well as clinical isolates of extended spectrum beta lactamase (ESBL)-positive E. coli and K. pneumoniae. AvBD-6 was more effective than AvBD-12 in neutralizing LPS and interacting with bacterial genomic DNA. Increasing bacterial concentration from 105 CFU/ml to 109 CFU/ml abolished AvBDs’ antimicrobial activity. Increasing NaCl concentration significantly inhibited AvBDs’ antimicrobial activity, but not the LPS-neutralizing function. Both AvBDs were mildly chemotactic for chicken macrophages and strongly chemotactic for CHO-K1 cells expressing chicken chemokine receptor 2 (CCR2). AvBD-12 at higher concentrations also induced chemotactic migration of murine immature dendritic cells (DCs). Disruption of disulfide bridges abolished AvBDs’ chemotactic activity. Neither AvBDs was toxic to CHO-K1, macrophages, or DCs.ConclusionsAvBDs are potent antimicrobial peptides under low-salt conditions, effective LPS-neutralizing agents, and broad-spectrum chemoattractant peptides. Their antimicrobial activity is positively correlated with the peptides’ net positive charges, inversely correlated with NaCl concentration and bacterial concentration, and minimally dependent on intramolecular disulfide bridges. In contrast, their chemotactic property requires the presence of intramolecular disulfide bridges. Data from the present study provide a theoretical basis for the design of AvBD-based therapeutic and immunomodulatory agents.

The Unusual Resistance of Avian Defensin AvBD7 to Proteolytic Enzymes Preserves Its Antibacterial Activity.

Defensins are frontline peptides of mucosal immunity in the animal kingdom, including birds. Their resistance to proteolysis and their ensuing ability to maintain antimicrobial potential remains questionable and was therefore investigated. We have shown by bottom-up mass spectrometry analysis of protein extracts that both avian beta-defensins AvBD2 and AvBD7 were ubiquitously distributed along the chicken gut. Cathepsin B was found by immunoblotting in jejunum, ileum, caecum, and caecal tonsils, while cathepsins K, L, and S were merely identified in caecal tonsils. Hydrolysis product of AvBD2 and AvBD7 incubated with a panel of proteases was analysed by RP-HPLC, mass spectrometry and antimicrobial assays. AvBD2 and AvBD7 were resistant to serine proteases and to cathepsins D and H. Conversely cysteine cathepsins B, K, L, and S degraded AvBD2 and abolished its antibacterial activity. Only cathepsin K cleaved AvBD7 and released Ile4-AvBD7, a N-terminal truncated natural peptidoform of AvBD7 that displayed antibacterial activity. Besides the 3-stranded antiparallel beta-sheet typical of beta-defensins, structural analysis of AvBD7 by two-dimensional NMR spectroscopy highlighted the restricted accessibility of the C-terminus embedded by the N-terminal region and gave a formal evidence of a salt bridge (Asp9-Arg12) that could account for proteolysis resistance. The differential susceptibility of avian defensins to proteolysis opens intriguing questions about a distinctive role in the mucosal immunity against pathogen invasion.

Construction of Recombinant Pichia pastoris Carrying a Constitutive AvBD9 Gene and Analysis of Its Activity.

Avian beta-defensin 9 (AvBD9) is a small cationic peptide consisting of 41 amino acids that plays a crucial rule in innate immunity and acquired immunity in chickens. Owing to its wide antibacterial spectrum, lack of a residue, and failure to induce bacterial drug resistance, AvBD9 is expected to become a substitute for conventional antibiotics in the livestock and poultry industries. Using the preferred codon of Pichia pastoris, the mature AvBD9 peptide was designed and synthesized, based on the sequence from GenBank. The P. pastoris constitutive expression vector pGHKα was used to construct a pGHKα-AvBD9 recombinant plasmid. Restriction enzyme digestion was performed using SacI and BglII to remove the ampicillin resistance gene, and the plasmid was electrotransformed into P. pastoris GS115. High-expression strains with G418 resistance were screened, and the culture supernatant was analyzed by Tricine-SDS-PAGE and western blot assay to identify target bands of about 6 kDa. A concentrate of the supernatant containing AvBD9 was used for determination of antimicrobial activity. The supernatant concentrate was effective against Escherichia coli, Salmonella paratyphi, Salmonella pullorum, Pseudomonas aeruginosa, Enterococcus faecalis, and Enterobacter cloacae. The fermentation product of P. pastoris carrying the recombinant AvBD9 plasmid was adjusted to 1.0 × 10(8) CFU/ml and added to the drinking water of white feather broilers at different concentrations. The daily average weight gain and immune organ indices in broilers older than 7 days were significantly improved by the AvBD9 treatment.

Administration of Poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) and Avian Beta Defensin as Adjuvants in Inactivated Inclusion Body Hepatitis Virus and its Hexon Protein-Based Experimental Vaccine Formulations in Chickens.

Inclusion body hepatitis (IBH) is one of the major infectious diseases adversely affecting the poultry industry of the United States and Canada. Currently, no effective and safe vaccine is available for the control of IBH virus (IBHV) infection in chickens. However, based on the excellent safety and immunogenic profiles of experimental veterinary vaccines developed with the use of new generation adjuvants, we hypothesized that characterization of vaccine formulations containing inactivated IBHV or its capsid protein hexon as antigens, along with poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) and avian beta defensin 2 (ABD2) as vaccine adjuvants, will be helpful in development of an effective and safe vaccine formulation for IBH. Our data demonstrated that experimental administration of vaccine formulations containing inactivated IBHV and a mixture of PCEP with or without ABD2 as an adjuvant induced significantly higher antibody responses compared with other vaccine formulations, while hexon protein-based vaccine formulations showed relatively lower levels of antibody responses. Thus, a vaccine formulation containing inactivated IBHV with PCEP or a mixture of PCEP and ABD2 (with a reduced dosage of PCEP) as an adjuvant may serve as a potential vaccine candidate. However, in order to overcome the risks associated with whole virus inactivated vaccines, characterization of additional viral capsid proteins, including fiber protein and penton of IBHV along with hexon protein in combination with more new generation adjuvants, will be helpful in further improvements of vaccines against IBHV infection.

Host Avian Beta-Defensin and Toll-Like Receptor Responses of Pigeons following Infection with Pigeon Paramyxovirus Type 1.

The high morbidity and mortality in pigeons caused by pigeon paramyxovirus type 1 (PPMV-1) highlights the need for new insights into the host immune response and novel treatment approaches. Host defense peptides (HDPs) are key components of the innate immune system. In this study, three novel avian β-defensins (AvBDs 2, 7, and 10) were characterized in pigeons and shown to possess direct antiviral activity against PPMV-1 in vitro. In addition, we evaluated the mRNA expression of these AvBDs and other immune-related genes in tissues of 2-month-old infected pigeons at 3 and 7 days postinfection. We observed that the expression of AvBD2 in the cecal tonsil, lungs, and proventriculus, as well as the expression of AvBD10 in the spleen, lungs, proventriculus, and kidneys, was upregulated in infected pigeons. Similarly, the expression of both Toll-like receptor 3 (TLR3) and TLR7 was increased in the spleen, trachea, and proventriculus, while TLR15 expression was increased only in the lungs of infected pigeons. In addition, inducible nitric oxide synthase (iNOS) expression was upregulated in the spleen, the bursa of Fabricius, the trachea, and the proventriculus of infected pigeons. Furthermore, we observed a high correlation between the expression of AvBD2 and the expression of either TLR7 or TLR15, as well as between AvBD10 expression and either TLR3 or TLR7 expression in respective tissues. The results suggest that PPMV-1 infection can induce innate host responses characterized by the activation of TLRs, particularly TLR3 and TLR7, AvBDs (2 and 10), and iNOS in pigeons.

Antimicrobial Genes Expression in Heterophils of Aseel

Salmonellosis is one of the most common infections in the chicken, and also the most important source of human food-borne infection. Heterophils are the predominate granulocyte in chicken expressing a variety of pathogen-associated molecular patterns (PAMPs) in killing the microbes via cationic antimicrobial peptides, such as beta-defensins that are capable of a broad spectrum of antimicrobial activity. Thus, the Toll-like receptors (TLRs) and beta-defensins are the major component of the heterophil antimicrobial arsenal. Aseel is considered to be robust and hardy in terms of disease resistance, but the immuno-genetic basis has not been investigated enough. The expression of the 14 Avian beta-defensins (AvBDs) and chicken Toll-like receptors (chTLRs) 1, 2, 3, 4, 5, 7, 15 and 21 were detectable in the heterophils. In spite of the efforts in prophylactic measures, vaccination and use of antibiotics, eradication of salmonellae from poultry stocks has not been very successful. An alternative approach chTLRs and beta defensin genes can be used as molecular markers in the selection of more resistant chicken lines.

Identification of three novel avian beta-defensins from goose and their significance in the pathogenesis of Salmonella

Here, we report the characterization of three avian β-defensins (AvBDs) from the goose, named anser_AvBD1, AvBD3, and AvBD6, respectively. All of anser_AvBDs exhibited broad antibacterial activity. In addition, the antibacterial activity of all of the AvBDs against Staphylococcus aureus and Proteus mirabilis decreased significantly in the presence of 100mM NaCl (P<0.01). None of the AvBDs showed hemolytic activity. In order to assess the significance of these anser_AvBDs in the infection of Salmonella enteritidis, mRNA expression of Toll-like receptor (TLR) 4 and anser_AvBDs in tissues of both control and infected geese was evaluated. We observed a significant up-regulation of TLR4, anser_AvBD1, 3, and 6 in some immune tissues evaluated, in response to S. enteritidis infection. These data demonstrated that TLR4 may serve a possible role in eliciting host immune responses to pathogens, and β-defensins may play a pivotal role in the host defense mechanisms of the goose.

Corrigendum to “Discovery and characterization of Coturnix chinensis avian β‐defensin 10, with broad antibacterial activity”. J. Pept. Sci. 2012; 18: 224–232

Some parts of the results in the article “Discovery and characterization of Coturnix chinensis avian b-defensin 10, with broad antibacterial activity” published in J. Pept. Sci. 2012; 18: 224–232, were published in an article in the Chinese Journal of Preventive Veterinary Medicine in Chinese (“Cloning and expression of quail avian beta-defensin 10 and the mRNA detection in quail tissues” 2011, 33, 552-556. doi:10.3969/j.issn.1008-0589.2011.07.12).

Correction: Identification, Expression and Activity Analyses of Five Novel Duck Beta-Defensins

The following two publications by our group should have been cited in this PLOS ONE article, the authors apologize for this oversight: Isolation, Characterization and Mechanism of Anti-viral Activity of Duck Avian Beta-defensin 16 Zhang K, Zhang M, Xin S, Han Z, Shao Y, Liu S and Ma D. Scientia Agricultura Sinica, 2012, 45(18): 3873-3882 (in Chinese) Isolation, characterization, and determination on bioactivity of duck avian beta-defensin 5 Zhang K, Lin L, Han Z, Shao Y, Liu S and Ma D. Microbiology China, 2011, 38(11): 1688-1697.(in Chinese) The sequences for Apl_AvBD16 and Apl_AvBD5 used for the study reported in PLOS ONE are identical with respect to AvBDs from the two reports in Chinese above, however, all the experiments for AvBD 16 and AvBD 5 were repeated to generate new datasets to be reported in the PLOS ONE article. Furthermore, the data for Apl_AvBD1, 3, and 6 reported in the PLOS ONE article were not included in the studies of the two Chinese publications above. The claim in the PLOS ONE article indicating that this reports the characterization of five novel defensins is inaccurate given that beta-defensin 16 and beta-defensin 5 had been already reported in the two previous publications above.