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The fecal virome comprises a complex diversity of eukaryotic viruses, phages and viruses that infect the host. However, little is known about the intestinal community of viruses that is present in wild waterfowl, and the structure of this community in wild ducks has not yet been studied. The fecal virome compositions of six species of wild dabbling ducks and one species of wild diving duck were thus analyzed. Fecal samples were collected directly from the rectums of 60 ducks donated by hunters. DNA and RNA virus particles were purified and sequenced using the MiSeq Illumina platform. The reads obtained from the sequencing were analyzed and compared with sequences in the GenBank database. Viral-related sequences from the Herpesviridae, Alloherpesviridae, Adenoviridae, Retroviridae and Myoviridae viral families showed the highest overall abundances in the samples. The virome analysis identified viruses that had not been found in wild duck feces and revealed distinct virome profiles between different species and between samples of the same species. This study increases our understanding of viruses in wild ducks as possible viral reservoirs and provides a basis for further studying and monitoring the transmission of viruses from wild animals to humans and disease outbreaks in domestic animals. [ABSTRACT FROM AUTHOR]

Chicken is widely favored by consumers because of some unique features. The leg muscles occupy an important position in the market. However, the specific mechanism for regulating muscle growth speed is not clear. In this experiment, we used Jinghai yellow chickens with different body weights at 300 days as research subjects. The chickens were divided into fast- and slow-growing groups, and we collected leg muscles after slaughtering for use in RNA-seq. After comparing the two groups, 87 differentially expressed genes (DEGs) were identified (fold change ≥ 2 and FDR < 0.05). The fast-growing group had 42 up-regulated genes and 45 down-regulated genes among these DEGs compared to the slow-growing group. Six items were significantly enriched in the biological process: embryo development ending in birth or egg hatching, chordate embryonic development, embryonic skeletal system development, and embryo development as well as responses to ketones and the sulfur compound biosynthetic process. Two significantly enriched pathways were found in the KEGG pathway analysis (P-value < 0.05): the insulin signaling pathway and the adipocytokine signaling pathway. This study provides a theoretical basis for the molecular mechanism of chicken growth and for improving the production of Jinghai yellow chicken. [ABSTRACT FROM AUTHOR]

Abstract The effects of coenzyme Q10 (CoQ10) has not yet been assessed for cryopreservation of rooster semen. The aim of this study was to evaluate the effect of different concentrations of CoQ10 in Lake extender for cryopreservation of rooster semen. The viability and apoptosis status, DNA fragmentation, abnormal morphology, motion parameters, membrane functionality, mitochondrial activity, acrosome integrity, lipid peroxidation, and fertility potential were evaluated after the freeze-thaw process. Semen samples were collected from ten roosters, twice a week, and then diluted in extender contained different concentrations of CoQ10 as follows: Lake without CoQ10 (control, Q 0), Lake containing 1 μM (Q 1), 2 μM (Q 2), 5 μM (Q 5), and 10 μM (Q 10) CoQ10. Supplementation of Lake with 1 and 2 μM CoQ10 resulted in greater sperm viability, total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, and fertility rate. Furthermore, the extent of lipid peroxidation in thawed spermatozoa treated with 1 and 2 μM CoQ10 was less than with the other groups. Different concentrations of CoQ10 had no effect on DNA fragmentation and sperm morphology. Results of the present study indicate that supplementation of Lake extender with 1 and 2 μM CoQ10 enhances the quality of rooster sperm after the freeze-thaw process. [ABSTRACT FROM AUTHOR]

Mucosal vaccine delivery systems have paramount importance for the induction of mucosal antibody responses. Two studies were conducted to evaluate immunogenicity of inactivated AIV antigens encapsulated in poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles (NPs). In the first study, seven groups of specific pathogen free (SPF) layer-type chickens were immunized subcutaneously at 7-days of age with different vaccine formulations followed by booster vaccinations two weeks later. Immune responses were profiled by measuring antibody (Ab) responses in sera and lachrymal secretions of vaccinated chickens. The results indicated that inactivated AIV and CpG ODN co-encapsulated in PLGA NPs (2x NanoAI+CpG) produced higher amounts of hemagglutination inhibiting antibodies compared to a group vaccinated with non-adjuvanted AIV encapsulated in PLGA NPs (NanoAI). The tested adjuvanted NPs-based vaccine (2x NanoAI+CpG) resulted in higher IgG responses in the sera and lachrymal secretions at weeks 3, 4 and 5 post-vaccination when immunized subcutaneously. The incorporation of CpG ODN led to an increase in Ab-mediated responses and was found useful to be included both in the prime and booster vaccinations. In the second study, the ability of chitosan and mannan coated PLGA NPs that encapsulated AIV and CpG ODN was evaluated for inducing antibody responses when delivered via nasal and ocular routes in one-week-old SPF layer-type chickens. These PLGA NPs-based and surface modified formulations induced robust AIV-specific antibody responses in sera and lachrymal secretions. Chitosan coated PLGA NPs resulted in the production of large quantities of lachrymal IgA and IgG compared to mannan coated NPs, which also induced detectable amounts of IgA in addition to the induction of IgG in lachrymal secretions. In both mucosal and subcutaneous vaccination approaches, although NPs delivery enhanced Ab-mediated immunity, one booster vaccination was required to generate significant amount of Abs. These results highlight the potential of NPs-based AIV antigens for promoting the induction of both systemic and mucosal immune responses against respiratory pathogens. [ABSTRACT FROM AUTHOR]

In this study, we describe an inter-laboratory collaborative ring trial validation of species-specific TaqMan real-time PCR assays for the detection of porcine- and chicken-derived materials in meat products. We comprehensively evaluated the performance of these assays in different environments and situations. This validation included the participation of thirteen laboratories across Europe and Asia. The results from the thirteen participating laboratories were analyzed to determine the specificity, accuracy, false positive rate, limit of detection (LOD), and probability of detection (POD) of the developed methods. These results indicated that the methods developed to detect porcine- and chicken-derived materials in meat products are robust and repeatable. The false positive and false negative rates were both 0%, and the LOD was determined to be five copies/reaction. The laboratory standard deviation (σL) was 0.30 for both detection methods, indicating that the developed methods are suitable for detection and identification of the porcine- and chicken-derived materials in meat products. [ABSTRACT FROM AUTHOR]

This study aimed to determine the impact of stocking density on the liver proteome and cecal microbiota of Peking ducks. A total of 1,200 21-day-old ducks were randomly assigned to 5 stocking density groups of 5, 6, 7, 8 and 9 ducks/m2, with 6 replicates for each group. At 40 days of age, duck serum and pectorals were collected for biochemical tests; liver and cecal contents of ducks were gathered for proteome and microbiota analysis, respectively. Serum MDA increased while pectorals T-AOC reduced linearly with enhancing stocking density. Duck lipid metabolism was altered under different stocking density as well. Serum LDL-C increased linearly with increasing stocking density. Proteome analysis revealed fatty acid biosynthesis proteins such as acyl-CoA synthetase family member 2 and fatty acid oxidation related proteins including acyl-CoA dehydrogenase long chain and acyl-coenzyme A oxidase were enriched in high stocking density group. Additionally, high stocking density increased oxidative response associated proteins such as DDRGK domain containing 1. Furthermore, increasing stocking density diminished proteins of anti-oxidant capacity including regucalcin and catalase. 16S rDNA analysis revealed that higher stocking density was accompanied with decreased microbial diversity, as well as depletion of anti-inflammatory bacterial taxa, including Bacteroidales, Butyricimonas and Alistipe. Besides, reduced bile acid metabolism-associated bacteria such as Ruminococcaceae, Clostridiales and Desulfovibrionaceae were found in the high-density group. Both proteome and 16S rDNA results showed inflammation and chronic liver disease trend in the high-density group, which suggests the involvement of the liver-gut axis in oxidative stress. [ABSTRACT FROM AUTHOR]

A multiplex quantitative PCR (qPCR) was developed and evaluated for the simultaneous detection of Salmonella spp., S. enterica serovar Typhimurium and S. enterica serovar Enteritidis in various (food) matrices. Early and fast detection of these pathogens facilitates effective intervention and prevents further distribution of contaminated food products on the market. Three primer and probe sets were designed to target the invA gene, the STM4200 gene, and the SEN1392 gene to detect and differentiate Salmonella spp., S. Typhimurium, and S. Enteritidis, respectively. The multiplex qPCR targeting these three genes was optimized for efficiency and linearity. By testing 225 Salmonella isolates and 34 non-Salmonella isolates from various sources the inclusivity and exclusivity were determined. The inclusivity of the multiplex qPCR was 100% for all Salmonella isolates, including 72 S. Typhimurium isolates, and 53 S. Enteritidis isolates. The exclusivity for Salmonella spp., S. Typhimurium, and S. Enteritidis was 100%, 94.6%, and 100%, respectively. No positive results were reported for non-Salmonella isolates. The limit of detection (LOD) for the qPCR was determined for the matrices poultry, minced meat, egg, herbs/spices, powdered milk, fish, animal feed, boot-socks with chicken feces and chicken down. LOD values for qPCR and the conventional culture methods were similar, except for the matrix boot-socks and down, for which the LOD for the conventional culture methods performed better than the qPCR method. In conclusion, the multiplex qPCR assay developed allows for rapid screening of Salmonella spp., S. Typhimurium, and S. Enteritidis in various (food) matrices. [ABSTRACT FROM AUTHOR]

Culture-based detection of Campylobacter can be affected by competing flora, temperature, incubation time, and presence of blood. The presence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in poultry has become one of the most common factors interfering with the detection of Campylobacter. In the present study, we evaluated potassium clavulanate (ESBL inhibitor) as a supplement in Bolton broth (C-Bolton broth) for enrichment and detection of Campylobacter. First, we determined growth kinetics of Campylobacter in the presence of different concentrations of ESBL E. coli in C-Bolton broth during enrichment. The effects of other factors such as incubation time, incubation temperature, and presence of blood on Campylobacter detection in C-Bolton broth were also investigated. The growth of Campylobacter co-cultured at a low concentration (2 and 4 log10 CFU/mL) of ESBL E. coli was similar to that of Campylobacter alone in C-Bolton broth, and Campylobacter co-cultured at a high concentration (6 and 8 log10 CFU/mL) of ESBL E. coli showed slower growth than the pure Campylobacter culture. The Campylobacter detection limit was 1 log10 CFU/mL when mixed with 2, 4, or 6 log10 CFU/mL of E. coli and 3 log10 CFU/mL when mixed with 8 log10 CFU/mL of E. coli after 48 h enrichment in the broth. Campylobacter detection from chicken feces and litter samples was not affected by incubation time, or presence of blood in the broth. A modified procedure of enrichment in C-Bolton broth at 37°C for 24 h without blood showed a significantly (P ≤ 0.05) higher detection rate and a lower false-negative rate than the ISO 10272–1:2006 method for Campylobacter detection from chicken feces and litter samples. In summary, the present study demonstrates the efficacy of Bolton broth supplemented with potassium clavulanate in the detection of Campylobacter mixed with ESBL E. coli, and an improved procedure to detect Campylobacter from chicken feces and litter samples. [ABSTRACT FROM AUTHOR]

Background: Food security is critical to achieving sustainable growth, poverty reduction, and political and economic stability. Livestock have the potential to improve the food security of smallholder households in developing countries, but livestock productivity is constrained by disease. The extent to which households adopt innovations such as vaccines impacts disease control; however, the behavioral and economic drivers underlying household decisions to adopt or forgo vaccination are not well understood. We address this gap with a study of adoption of Newcastle disease (ND) vaccines by chicken-owning households in Tanzania. Methods: A cross-sectional survey was administered to 535 households owning indigenous chickens in Arusha, Singida, and Mbeya regions in Tanzania. We measured potential predictors of ND vaccine adoption including knowledge, attitudes, and practices. Logistic regression was used to identify predictors correlated with three stages of household adoption: awareness of ND vaccines, previous vaccination, and recent vaccination (within four months) consistent with veterinary guidelines. Results: Eighty percent of households were aware of ND vaccines, 57% had previously vaccinated, and 26% had recently vaccinated. Knowing someone who vaccinated increased the odds of a household previously vaccinating [adjusted odds ratio (AOR): 1.32, 95% CI: 1.1–1.5]. Larger flock size was also associated with higher odds of previous vaccination (AOR: 1.03 for a one chicken increase, 95% CI: 1.01–1.05). Usage of traditional medicine decreased the odds of previously vaccination (AOR: 0.58, 95% CI: 0.36–0.95). Conclusion: Our findings suggest that encouraging the flow of professional-level knowledge within the community by vaccine adopters is a strategy to increase vaccine adoption. Enhancing local chicken productivity through increased vaccine coverage would strengthen a key smallholder household resource for food and economic security. [ABSTRACT FROM AUTHOR]

We describe and test an obstacle-detection system for small, lake-deployed autonomous surface vehicles (ASVs) that relies on a low-cost, consumer-grade camera and runs on a single-board computer. A key feature of lakes that must be accounted for is the frequent presence of the shoreline in images as well as the land-sky boundary. These particularities, along with variable weather conditions, result in a wide range of scene variations, including the possible presence of glint. The implemented algorithm is based on two main steps. First, possible obstacles are detected using an innovative gradient-based image processing algorithm developed especially for a camera with a low viewing angle to the water (i.e., the situation for a small ASV). Then, true and false positives are differentiated using correlation-based multi-frame analysis. The algorithm was tested extensively on a small ASV deployed in Lake Geneva. Under operational conditions, the algorithm processed 640×480-pixel images from a Raspberry Pi Camera at about 3—4 Hz on a Raspberry Pi 3 Model B computer. The present algorithm demonstrates that single-board computers can be used for effective and low-cost obstacle detection systems for ASVs operating in variable lake conditions. [ABSTRACT FROM AUTHOR]

The transition to using dual-purpose chickens is an alternative to killing male hatchlings of high performance egg-laying chickens. This study aimed to compare the gastrointestinal tract of a recently developed genetic line of dual purpose male chicken, Lohmann Dual (LD), with that of a broiler line, Ross 308. Eighty birds from each line were grown until they reached an average body weight 2000 g (5 weeks for Ross and 9 for LD birds). Six birds of each line were sampled weekly. Body weight (BW), normalized mass of gastrointestinal segments and relative length of intestine were determined. Histologically the villus height, epithelium height, crypt depth, mucosal enlargement factor and the tunica muscularis thickness were measured in jejunum and ileum. Data were regressed against body weight and genetic line. Jejunal enterocyte microvilli and junctional complexes length were measured. Normalized mass and relative length of the gastrointestinal segments were greater in LD birds than in Ross birds at all ages. After day 7 these decreased steadily over the lifetime of the birds in both genetic lines. The growth curves of the gastrointestinal segments of the LD birds were similar to those of the Ross birds. In birds of the same BW, LD birds had a significantly heavier gizzard, shorter intestine, higher jejunal villi, thicker ileal tunica muscularis and smaller ileal mucosal enlargement factor than were found in Ross birds. The large gizzard in LD chickens presumably increases the degree of food processing and enhances availability of nutrients in the orad part of the intestine leading to a lower nutrient concentration and a smaller absorption surface area in the ileum of the LD compared to the Ross chickens. The anatomical differences between the two lines are important criteria for further selection and should be considered in their feeding management. [ABSTRACT FROM AUTHOR]

The objective of this study was to identify potential mRNA expression changes in chicken livers associated with environmental exposure to dichloro-diphenyl-trichloroethane (DDT) and its metabolites (DDTs). In particular, we focused on genes relating to the immune system and metabolism. We analyzed liver samples from free-ranging chickens in KwaZulu-Natal, South Africa, for contamination by DDTs. This area predominantly uses DDT in its malaria control program, and homes are sprayed annually with the pesticide. Genes relating to the immune system and metabolism were selected as potential genetic biomarkers that could be linked to higher contamination with DDTs. RT-qPCR analysis on 39 samples showed strong correlations between DDTs contamination and mRNA expression for the following genes: AvBD1, AvBD2, AvBD6 and AvBD7 (down-regulated), and CYP17, ELOVL2 and SQLE (up-regulated). This study shows for the first time interesting and significant correlations between genetic material collected from environmentally-exposed chickens and mRNA expression of several genes involved in immunity and metabolism. These findings show the usefulness of analysis on field samples from a region with high levels of environmental contamination in detecting potential biomarkers of exposure. In particular, we observed clear effects from DDT contamination on mRNA expression of genes involved in immune suppression, endocrine-disrupting effects, and lipid dysregulation. These results are of interest in guiding future studies to further elucidate the pathways involved in and clinical importance of toxicity associated with DDT exposure from contaminated environments, to ascertain the health risk to livestock and any subsequent risks to food security for people. [ABSTRACT FROM AUTHOR]

Red Junglefowls (RJFs), the wild progenitor of modern day chickens (DCs), are believed to be in genetic endangerment due to introgression of domestic genes through opportunistic matings with domestic or feral chickens. Previous studies from India reported rare hybridization of RJFs in the wild. However, RJF population genetic structure, pattern of gene flow and their admixture with DC populations are poorly understood at the landscape level. We conducted this study with a large sample size, covering the predicted natural distribution range of RJFs in India. We documented strong evidence of directional gene flow from DCs to free-ranging wild RJFs, with the Northeastern RJF population exhibiting the most genetic variants in their nuclear and mitochondrial genomes, indicating it to be the ancestral population from which early radiation may have occurred. The results provide evidence that landscape features do not act as a barrier to gene flow and the distribution pattern could not be explored due to physical sharing or exchange of wild birds in the past when forests were continuous across RJF range in India. [ABSTRACT FROM AUTHOR]

The pancreatic secretion of insulin, a key endocrine regulator of metabolism and growth, can be greatly influenced by the gut-derived incretin hormones, namely by GIP (Glucose-dependent Insulinotropic Peptide) and GLP-1 (Glucagon-like Peptide 1). As insulin is a major stimulator of growth, affecting its producion may be of special importance in food-producing livestock. The aim of the present study was to investigate novel ways of modulating incretin and insulin homeostasis in chickens and rabbits by nutrition, e.g. by oral butyrate application, also studying the mechanisms of incretin action in both species as a comparative approach. Acute oral butyrate challenge significantly decreased plasma GIP levels by approx. 40% in both species: significant interactions of butyrate exposure and incubation time were found in both chickens (P = 0.038 and P = 0.034 at 30 and 60 min following butyrate ingestion [1.25 g/kg BW], respectively) and rabbits (P = 0.036 and P = 0.039 at 30 and 60 min after butyrate ingestion [0.25 g/kg BW], respectively), while plasma GLP-1, insulin and glucose concentrations remained unaffected by butyrate in both species over time. These results are in contrast to butyrate’s stimulating effect on both incretin and insulin secretion in mice, indicating specific, species-dependent differences even among mammalian species. Further, based on the analyzed correlations between the measured endocrine parameters (regardless of the butyrate exposure), it can be assumed that incretins may regulate pancreatic insulin release in rabbits on a partly different way compared to mice, humans and chickens. [ABSTRACT FROM AUTHOR]

Wetland loss and degradation have been extensive across the world, especially in California’s Central Valley where over 90% of the natural wetlands have been converted to agricultural and urban uses. In the Central Valley today, a much smaller network of managed wetlands and flooded agricultural fields supports almost five million waterfowl and half a million shorebirds. Over 50% of waterbird habitat in the Central Valley is provided by flooded agricultural land, primarily rice (Oryza sativa). Each year non-breeding waterbird habitat decreases in the late winter as flooded agricultural fields are drained after waterfowl hunting season in late-January to prepare for the next crop. This study evaluated a practice called ‘variable drawdown’ that involves delaying the removal of water from rice fields by 1, 2, and 3 weeks to extend the availability of flooded habitat later into February and March. We studied waterbird response to variable drawdown in 2012 and 2013 at twenty rice farms throughout the northern half of the Central Valley. The staggered drawdown created a mosaic of water depths throughout the six-week study period. The 3-week delay in drawdown supported more dabbling ducks than earlier drawdowns in the first half of the study and more shorebirds and long-legged wading birds during the second half of the study. The timing of highest use of each drawdown treatment differed for each waterbird guild; dabbling ducks, geese and swans benefited at the beginning, then long-legged wading birds, followed by shorebirds. Despite the presence of appropriate water depths for shorebirds across the treatments during the entire study period, shorebird densities were highest near the end of the study when the 3-week-delayed drawdown was providing the majority of the habitat on the landscape. This suggests that shorebirds may have concentrated in our study fields due to decreasing availability of shallow water habitat elsewhere. The practice of variable drawdown successfully extended the availability of waterbird habitat provided by post-harvest flooded rice fields later into winter. [ABSTRACT FROM AUTHOR]

Avian leukosis virus subgroup J (ALV-J) can induce myeloid tumors and hemangiomas in chickens and causes severe economic losses with commercial layer chickens and meat-type chickens. Here, we generated ribominus RNA sequencing data from three normal chicken spleen tissues and three ALV-J-infected chicken spleen tissues. Structure analysis of transcripts showed that, compared to mRNAs and lncRNAs, chicken circRNAs shared relatively shorter transcripts and similar GC content. Differentially expression analysis showed 152 differentially expressed circRNAs with 106 circRNAs up regulated and 46 circRNAs down regulated. Through comparing differentially expressed circRNA host genes and mRNAs and performed ceRNA network analysis, we found several tumor or immune-related genes, in which, there were four genes existed in both differentially expressed mRNAs and circRNA host genes (Dock4, Fmr1, Zfhx3, Ralb) and two genes (Mll, Aoc3) involved in ceRNA network. We further characterized one exon-intron circRNA derived from HRH4 gene in the ceRNA network, termed circHRH4, which is an abundant and stable circRNA expressed in various tissues and cells in chicken and localizes in cytoplasm. Our results provide new insight into the pathology of ALV-J infection and circRNAs may also mediate tumorigenesis in chicken. [ABSTRACT FROM AUTHOR]

Similar to mammals, the avian heart is a muscular pump responsible for distributing blood throughout the body. However, a larger relative heart weight to body weight (RHW/BW) was reported in birds than mammals. Furthermore, it has been revealed that smaller birds specially flying one have relatively larger hearts (relative to body mass) than larger birds (non-flying). Although this fact, there are no literatures regarding comparative descriptive variations of the heart in flying and non-flying birds that could adapt various functions, such as flying and running. Therefore, this study aimed to conduct a comparative anatomical features and morphometrical measurments between the hearts of pigeon and fowl. The present study also analyzed the immunohistochemical expression of Troponin T (TnT) as one of the major regulator of striated muscle contraction. The study revealed significant larger RHW/BW in pigeon heart. Furthermore, the hearts of both fowl and pigeon are directed caudoventrally in the cranial part of thoracoabdominal cavity, however, a more oblique position nearly parallel to the sternum was observed in pigeon's hearts. Interestingly, the mean morphometric values (length of both cranial and caudal borders, diameter of the base, and thickness of both right and left ventricular wall and interventricular septum) of the pigeon hearts showed significant higher differences than that of fowl. Additionally, more expression of TnT was observed in pigeon hearts. Therefore, our findings suggest that such morphological and immunological varities are possibly essential factors for providing intense ventricular contraction of the pigeon's heart to cover the high metabolic requirments for flight. [ABSTRACT FROM AUTHOR]

Background: Chagas disease (CD) is a life-threatening illness caused by the protozoan parasite Trypanosoma cruzi, which is transmitted by triatomine bugs. Triatomine bugs inhabit poorly constructed homes that create multiple hiding spots for the bugs. Modifying the actual structure of a home, along with the homeowners’ practices, can reduce triatomine infestation. This research was designed to collect culturally-relevant information to develop a health campaign to decrease risk of CD transmission by promoting home maintenance and better hygiene in rural communities of southern Ecuador. Methods and main findings: The Health Belief Model (HBM) guided focus group discussions and the interpretation of the results. Four focus groups ranging from 4 to 10 participants were conducted between May and June 2014 in three communities of Loja province in Southern Ecuador. A thematic analysis was used to identify within the data related to perceptions of susceptibility, severity, benefits, barriers and self-efficacy related to CD and its prevention. The results provide clear guidance for the development of Chagas-prevention messages. Conclusion: Data obtained emphasize the importance of standardizing messages presented to the communities for CD prevention. Messages should provide more information on the protective nature of the behaviors promoted for CD prevention; overcoming barriers such as cost and convenience, and build on facilitating factors, including community members’ interest on quality of life, protection of their families, and relationship with the land. [ABSTRACT FROM AUTHOR]

Among the organic acids, lauric acid has shown a high level of in vitro activity against Campylobacter jejuni. The prevalence and intensity of C. jejuni excretion at slaughter often becomes lower with increasing age. In higher-aged broilers on organic farms which often use other breeds, in turn, the prevalence of C. jejuni is sometimes higher at slaughter. The question then arises as to whether a diet with higher lauric acid concentrations, the age alone or the genetic breed might have an effect in the spread and intensity of an experimental C. jejuni infection in vivo. Therefore, two complete diets with or without 2% lauric acid from palm kernel fatty acids were offered to 450 chickens (ten subgroups à 15 birds, repetitions: n = 3) of two broiler and two layer breeds (Ross 308, Hubbard JA 757, Lohmann Dual and Lohmann Brown-Classic). All breeds were reared for 42 days, Lohmann Brown-Classic also for about 98 days. Twenty-one days before dissection, three seeder birds per subgroup were orally infected with a 1 mL inoculum of C. jejuni (4.46±0.35 log10 CFU/mL). Qualitative detection of C. jejuni in cloacal swabs was performed at days 2, 4, 7, 14 after inoculation and at dissection in all birds. Quantitative detection was performed on excreta samples of seeder birds at days 2, 11 and 17 after experimental challenge and on caecal samples of all birds at dissection. Two days after experimental inoculation, C. jejuni prevalence was higher in control birds without lauric acid supplementation (48.9% vs. 39.6%; P = 0.0462). Depending on age, two days after inoculation the C. jejuni prevalence in young Lohmann Brown-Classic chickens was significantly lower (37.8% vs. 61.1%) whereas at dissection it was higher (99% vs. 67%). At day 2 after inoculation C. jejuni counts in the excreta of young Lohmann Brown-Classic were lower in comparison to those in old ones (log10 CFU/g: 3.30±2.68 vs. 5.24±1.56). Eleven (log10 CFU/g: 5.14±1.13 vs. 4.16±0.82) and 17 days after inoculatioin (log10 CFU/g: 3.77±2.02 vs. 1.72±1.87) it was the reverse situation. At dissection, the carriage of C. jejuni in caecal content was higher in younger than in older birds (log10 CFU/g: 8.57±0.46 vs. 6.66±1.43). An effect of genetic breed on C. jejuni prevalence was seen at dissection, this being lowest in Lohmann Dual chickens (91% vs. 98.9% in other breeds). At d 17 after challenge, C. jejuni counts in the excreta of young Lohmann Brown-Classic were lower in comparison to Ross 308 and Hubbard JA 757 (log10 CFU/g: 3.77±2.02 vs. 5.21±0.85 and 5.62±0.90). Lohmann Dual chickens showed an intermediary excretion, this being only significant lower compared to Hubbard JA 757 (log10 CFU/g: 4.31±0.89). In summary, the effect of lauric acid is limited to the initial phase after experimental inoculation. A higher age at infection seems to lead to a more rapid limitation of the infection. The excretion of C. jejuni appears to decrease more rapidly in layer breeds than in broiler lines after experimental inoculation. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is a widespread infectious disease, leading to a major health and economic burden. Chickens are considered as the most common infection source for humans. Campylobacter mainly multiplies in the mucus layer of their caeca. No effective control measures are currently available, but passive immunisation of chickens with pathogen-specific maternal IgY antibodies, present in egg yolk of immunised chickens, reduces Campylobacter colonisation. To explore this strategy further, anti-Campylobacter nanobodies, directed against the flagella and major outer membrane proteins, were fused to the constant domains of chicken IgA and IgY, combining the benefits of nanobodies and the effector functions of the Fc-domains. The designer chimeric antibodies were effectively produced in leaves of Nicotiana benthamiana and seeds of Arabidopsis thaliana. Stable expression of the chimeric antibodies in seeds resulted in production levels between 1% and 8% of the total soluble protein. These in planta produced antibodies do not only bind to their purified antigens but also to Campylobacter bacterial cells. In addition, the anti-flagellin chimeric antibodies are reducing the motility of Campylobacter bacteria. These antibody-containing Arabidopsis seeds can be tested for oral passive immunisation of chickens and, if effective, the chimeric antibodies can be produced in crop seeds. [ABSTRACT FROM AUTHOR]

The domesticated phenotype is a set of behavioural, morphological and physiological traits that is common for domesticated species. Previous research has found that selection for tameness only can give rise to correlated selection responses that resemble the domesticated phenotype. It has therefore been suggested that tameness may drive the domesticated phenotype as correlated traits. We selected Red Junglefowl for divergent levels of fear of human for eight generations and assessed possible correlated selection responses in other behaviours in semi-natural settings. Behavioural studies were carried out on birds from generations six to eight, and at different ages, in order to study possible effects on general activity, social behaviour and male courtship behaviour. We found that the differences between selection lines changed with age. Adult low fear birds were generally more active and high fear males showed a more intense courtship behaviour. Our study shows that several behaviours can be modified through correlated selection responses by selection on reduced fear of humans only, emphasising the putative role of tameness as a driver of domestication related phenotypes. [ABSTRACT FROM AUTHOR]

Diseases in introduced broilers can possibly spill over to wild birds on the Galapagos. Knowledge about the current burden of exposure to pathogens in broilers on the Galapagos is very limited. The objective of the study reported here was to measure the burden of exposure to infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (MG), and intestinal parasites in a sample of broiler chickens on 13 farms on Santa Cruz Island and San Cristobal Island in July 2017. Blood serum samples were tested for detection of antibodies to IBDV, IBV, NDV, and MG by using an IDEXX Enzyme-linked Immunosorbent Assay. In addition, fecal samples and pen bedding environmental samples were processed and analyzed for diagnosis of intestinal parasite eggs under a compound light microscope. The frequency of seropositive broilers to IBDV was 74/130 or 56% (95% CI = 48, 65%), to IBV was 27/130 or 20% (14, 28%), and to NDV was 1/130 or 0.7% (0.1, 4%). All broilers tested negative to MG antibodies. Eimeria spp. infection was common in study broilers. Finally, we observed interaction between broiler chickens and wild birds (finches) inside broiler pens, as well as the presence of backyard chickens inside property limits of study farms. This study produced evidence that exposure to IBDV, IBV, and intestinal parasites in broilers on Santa Cruz Island and San Cristobal Island is important. Study results are relevant because (i) they provide new baseline data on the burden of exposure to avian pathogens in broiler farms, (ii) justify the need to verify standard operating procedures in hatcheries that supply (non-vaccinated) day-old chicks to the Galapagos and (iii) to implement enhanced biosecurity standards on broiler chicken farms to mitigate risk of disease transmission between broilers, backyard poultry, and wild birds on the Galapagos. [ABSTRACT FROM AUTHOR]

The spatial spread of highly pathogenic avian influenza (HPAI) H5N2 during the 2015 outbreak in the U.S. state of Minnesota was analyzed through the estimation of a spatial transmission kernel, which quantifies the infection hazard an infectious premises poses to an uninfected premises some given distance away. Parameters were estimated using a maximum likelihood method for the entire outbreak as well as for two phases defined by the daily number of newly detected HPAI-positive premises. The results indicate both a strong dependence of the likelihood of transmission on distance and a significant distance-independent component of outbreak spread for the overall outbreak. The results further suggest that HPAI spread differed during the later phase of the outbreak. The estimated spatial transmission kernel was used to compare the Minnesota outbreak with previous HPAI outbreaks in the Netherlands and Italy to contextualize the Minnesota transmission kernel results and make additional inferences about HPAI transmission during the Minnesota outbreak. Lastly, the spatial transmission kernel was used to identify high risk areas for HPAI spread in Minnesota. Risk maps were also used to evaluate the potential impact of an early marketing strategy implemented by poultry producers in a county in Minnesota during the outbreak, with results providing evidence that the strategy was successful in reducing the potential for HPAI spread. [ABSTRACT FROM AUTHOR]

The calcium binding protein ALG-2 is upregulated in several types of cancerous tissues and cancer cell death may be a consequence of ALG-2 downregulation. Novel research suggests that ALG-2 is involved in membrane repair mechanisms, in line with several published studies linking ALG-2 to processes of membrane remodeling and transport, which may contribute to the fitness of cells or protect them from damage. To investigate the involvement of ALG-2 in cell recovery after membrane damage we disrupted the PDCD6 gene encoding the ALG-2 protein in DT-40 cells and exposed them to electroporation. ALG-2 knock-out cells were more sensitive to electroporation as compared to wild type cells. This phenotype could be reversed by reestablishing ALG-2 expression confirming that ALG-2 plays an important role in cell recovery after plasma membrane damage. We found that overexpression of wild type ALG-2 but not a mutated form unable to bind Ca2+ partially protected HeLa cells from digitonin-induced cell death. Further, we were able to inhibit the cell protective function of ALG-2 after digitonin treatment by adding a peptide with the ALG-2 binding sequence of ALIX, which has been proposed to serve as the ALG-2 downstream target in a number of processes including cell membrane repair. Our results suggest that ALG-2 may serve as a novel therapeutic target in combination with membrane damaging interventions. [ABSTRACT FROM AUTHOR]

Scalable production of avian cell lines exhibits a valuable potential on therapeutic application by producing recombinant proteins and as the substrate for virus growth due to the special glycosylation occurs in avian species. Chicken primordial germ cells (cPGCs), a germinal pluripotent avian cell type, present the ability of self-renewal, an anchorage-independent cell growth and the ability to be genetically modified. This cell type could be an interesting bioreactor system for industrial purposes. This study sought to establish an expandable culture system with defined components for three-dimensional (3D) culture of cPGCs. cPGCs were cultured in medium supplemented with the functional polymer FP003. Viscoelasticity was low in this medium but cPGCs did not sediment in culture and efficiencies of space and nutrient utilization were thus enhanced and consequently their expansion was improved. The total number of cPGCs increased by 17-fold after 1 week of culture in 3D-FAot medium, an aseric defined medium containing FP003 polymer, FGF2 and Activin A as growth factors and Ovotransferrin as protein. Moreover, cPGC cell lines stably expressed the germline-specific reporter VASA:tdTOMATO, as well as other markers of cPGCs, for more than 1 month upon culture in 3D-FAot medium, indicating that the characteristics of these cells are maintained. In summary, this novel 3D culture system can be used to efficiently expand cPGCs in suspension without mechanical stirring, which is available for long-term culture and no loss of cellular properties was found. This system provides a platform for large-scale culture of cPGCs. [ABSTRACT FROM AUTHOR]

Infectious bronchitis virus (IBV) affects poultry respiratory, renal and reproductive systems. Currently the efficacy of available live attenuated or killed vaccines against IBV has been challenged. We designed a novel IBV vaccine alternative using a highly innovative platform called Self-Assembling Protein Nanoparticle (SAPN). In this vaccine, B cell epitopes derived from the second heptad repeat (HR2) region of IBV spike proteins were repetitively presented in its native trimeric conformation. In addition, flagellin was co-displayed in the SAPN to achieve a self-adjuvanted effect. Three groups of chickens were immunized at four weeks of age with the vaccine prototype, IBV-Flagellin-SAPN, a negative-control construct Flagellin-SAPN or a buffer control. The immunized chickens were challenged with 5x104.7 EID50 IBV M41 strain. High antibody responses were detected in chickens immunized with IBV-Flagellin-SAPN. In ex vivo proliferation tests, peripheral mononuclear cells (PBMCs) derived from IBV-Flagellin-SAPN immunized chickens had a significantly higher stimulation index than that of PBMCs from chickens receiving Flagellin-SAPN. Chickens immunized with IBV-Flagellin-SAPN had a significant reduction of tracheal virus shedding and lesser tracheal lesion scores than did negative control chickens. The data demonstrated that the IBV-Flagellin-SAPN holds promise as a vaccine for IBV. [ABSTRACT FROM AUTHOR]

Salmonella enterica subsp. enterica serovar Heidelberg (S. Heidelberg) is a clinically-important serovar linked to food-borne illness, and commonly isolated from poultry. Investigations of a large, multistate outbreak in the USA in 2013 identified poultry litter (PL) as an important extra-intestinal environment that may have selected for specific S. Heidelberg strains. Poultry litter is a mixture of bedding materials and chicken excreta that contains chicken gastrointestinal (GI) bacteria, undigested feed, feathers, and other materials of chicken origin. In this study, we performed a series of controlled laboratory experiments which assessed the microevolution of two S. Heidelberg strains (SH-2813 and SH-116) in PL previously used to raise 3 flocks of broiler chickens. The strains are closely related at the chromosome level, differing from the reference genome by 109 and 89 single nucleotide polymorphisms/InDels, respectively. Whole genome sequencing was performed on 86 isolates recovered after 0, 1, 7 and 14 days of microevolution in PL. Only strains carrying an IncX1 (37kb), 2 ColE1 (4 and 6kb) and 1 ColpVC (2kb) plasmids survived more than 7 days in PL. Competition experiments showed that carriage of these plasmids was associated with increased fitness. This increased fitness was associated with an increased copy number of IncX1 and ColE1 plasmids. Further, all Col plasmid-bearing strains had hotspot mutations in 37 loci on the chromosome and in 3 loci on the IncX1 plasmid. Additionally, we observed a decrease in susceptibility to tobramycin, kanamycin, gentamicin, neomycin and fosfomycin for Col plasmid-bearing strains. Our study demonstrates how positive selection from poultry litter can change the evolutionary path of S. Heidelberg. [ABSTRACT FROM AUTHOR]

Mycoplasma gallisepticum is a serious pathogen for poultry that causes chronic respiratory disease in chickens. Increased embryonic mortality, as well as reduced weight gain and egg production have been found in infected chickens, which can lead to considerable economic losses in poultry production. Increased antibiotic resistance compromises the use of tetracyclines, macrolides and quinolones in the farm environment. In the present study, danofloxacin concentrations were simulated below the MIC99, between the MIC99 and MPC (the mutant prevention concentration), and above the MPC in an in vitro dynamic model against M. gallisepticum. The relationship between the simulated danofloxacin pharmacokinetics, pharmacodynamics (PK/PD) parameters and development of resistance for M. gallisepticum was explored based on the available data obtained from various dosing regimens in the in vitro model. Danofloxacin concentration, counts of viable cell and susceptibility were determined during the experiment. The mutations in gyrA, gyrB, parC and parE as well as efflux pumps were examined. The MIC of danofloxacin against M. gallisepticum was increased when drug concentrations were between the lower and upper boundaries of the mutant selection window. The upper boundary of the selection window in vitro was estimated as a Cmax/MPC value of 1. The lower boundary was estimated as Cmax/MPC value of 0.05. Both in terms of the MIC and resistance frequency, M. gallisepticum resistance was developed when danofloxacin concentrations fell inside the mutant selection window (ratios of Cmax to MPC between 0.05 and 1). The single mutation in gyrA (Ser-83→Arg) was found in all mutants, while double mutations in gyrA and parC (Ala-64→Ser) were observed only in the mutant with the highest MIC. In addition, no change of susceptibility in the mutants was observed in the presence of reserpine and carbonyl cyanide 3-chlorophenylhydrazone (CCCP). This suggested that ATP-binding cassette superfamily (ABC transporter) and major facilitator superfamily (MFS transporter) did not play a role in danofloxacin efflux. [ABSTRACT FROM AUTHOR]

Background: This cross-sectional study determined the prevalence of Salmonella spp. and their serotypes on dressed chicken sold at retail outlets in Trinidad. The study also investigated the risk factors for contamination of dressed carcasses by Salmonella spp. at cottage poultry processor outlets where chickens are slaughtered and processed for sale. Methods: A total of 133 dressed, whole chickens and 87 chicken parts from 44 cottage poultry processors and 36 dressed, whole chickens and 194 chicken parts from 46 supermarket outlets were randomly collected throughout the country. Isolation and identification of Salmonella spp. were performed using standard bacteriological techniques. Serotyping was performed by a regional reference laboratory. Results: The prevalence of Salmonella spp. in chicken carcasses sampled from cottage poultry processors and supermarkets was 20.5% and 8.3% respectively (p <0.001). The frequency of isolation of Salmonella spp. at cottage poultry processors was 22.4%, 23.0%, 7.1%, and 10.0% for non-chilled whole chicken, non-chilled chicken parts, chilled whole chicken and chilled chicken parts respectively. Fresh, non-chilled chicken (22.6%) yielded a higher frequency of isolation of Salmonella spp. than chilled chickens (8.3%). For supermarket samples, the frequency of isolation of Salmonella spp. was 19.0%, 8.1%, 0.0% and 7.6% for chilled whole chickens, chill chicken parts, frozen whole chicken and frozen chicken parts respectively. The swab method of sampling yielded a statistically significantly (p = 0.029) higher frequency (3.2%) of Salmonella spp. than the rinse method (1.6%). The predominant serotypes isolated were Kentucky (30.9%) and Javiana (22.7%). Use of chilled water-bath to cool carcasses was the only risk factor significantly (p = 0.044) associated with isolation of Salmonella spp. Conclusion: Raw chicken carcasses purchased from cottage poultry processors pose a significantly higher risk of contamination with Salmonella spp. than those sold at supermarkets. [ABSTRACT FROM AUTHOR]

Inosine monophosphate (IMP) is a key factor affecting the fleshy flavor of meat; meanwhile, the free-range mode is an efficient strategy to improve muscular IMP content. To assess expression differences in IMP metabolism-related genes under different feeding patterns, Illumina Nextseq 500 sequencing was used to catalog the global gene expression profiles of muscle samples from Lueyang black-bone chicken under free-range and caging conditions. A total of 15510 unigenes were assembled, with 13423 (86.54%) and 6088 (39.25%) unigenes correctly annotated in the GO and KOG databases, respectively. Next, the "purine metabolism" pathway in the "nucleotide metabolism group" was assessed in depth. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, we retrieved 172 nucleotide- and 5 purine- metabolism related genes that were differentially expressed in muscle samples from free-range and caged chickens. At 60-day-old, AMPD1, NT5C1A and ENTPD8 showed higher levels in the free-range group, while only ENTPD8 was upregulated in 120-day-old chickens. In addition, GART, GARS and ADSL in free-range chickens showed higher levels compared with caged animals. Furthermore, IMPDH levels in free-range chicken were lower than those of caged chicken. Real-time quantitative polymerase chain reaction (qPCR) was used to validate the above findings. These results revealed a set of differentially expressed genes potentially related to IMP metabolism in chicken under different breeding modes, providing novel insights into controlling IMP levels in chicken meat. [ABSTRACT FROM AUTHOR]

Kinase-mediated protein phosphorylation is a central mechanism for regulation of cellular responses and phenotypes. While considerable information is available regarding the evolutionary relationships within the kinase family, as well as the evolutionary conservation of phosphorylation sites, each aspect of this partnership is typically considered in isolation, despite their clear functional relationship. Here, to offer a more holistic perspective on the evolution of protein phosphorylation, the conservation of protein phosphorylation sites is considered in the context of the conservation of the corresponding modifying kinases. Specifically, conservation of defined kinase-phosphorylation site pairings (KPSPs), as well as of each of the component parts (the kinase and the phosphorylation site), were examined across a range of species. As expected, greater evolutionary distance between species was generally associated with lower probability of KPSP conservation, and only a small fraction of KPSPs were maintained across all species, with the vast majority of KPSP losses due to the absence of the phosphorylation site. This supports a model in which a relatively stable kinome promotes the emergence of functional substrates from an evolutionarily malleable phosphoproteome. [ABSTRACT FROM AUTHOR]

A key step in understanding the evolution of human language involves unravelling the origins of language’s syntactic structure. One approach seeks to reduce the core of syntax in humans to a single principle of recursive combination, , for which there is no evidence in other species. We argue for an alternative approach. We review evidence that beneath the staggering complexity of human syntax, there is an extensive layer of nonproductive, nonhierarchical syntax that can be fruitfully compared to animal call combinations. This is the essential groundwork that must be explored and integrated before we can elucidate, with sufficient precision, what exactly made it possible for human language to explode its syntactic capacity, transitioning from simple nonproductive combinations to the unrivalled complexity that we now have. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is a leading foodborne zoonosis worldwide, and is frequently associated with handling and consumption of poultry meat. Various studies indicate that Campylobacter causes a substantial human disease burden in low to middle-income countries, but data regarding the organism’s epidemiology in countries like Kenya are scarce. In sub-Saharan Africa, 3.8 million deaths of children under-5 years of age are reported annually. Of those, 25% are caused by diarrheal diseases, and Campylobacter is one of the most frequently isolated bacteria from diarrheic children. With the growth of urban conglomerates, such as Kenya’s capital, Nairobi, changes in diets, food production systems, and retailing dynamics, it is likely that exposure and susceptibility to this pathogen will change. Therefore, the importance of Campylobacter disease burden in Kenya may increase further. The objectives of this study were: 1) to determine the prevalence of Campylobacter spp. in Nairobi’s small-scale chicken farms and meat retailers, and 2) to identify potential risk factors associated with its presence in those sites. The prevalence data provides the first detailed baseline for this pathogen in the urban Kenyan context. The risk factors provide context-specific insights for disease managers. A cross-sectional study of broiler, indigenous chicken farms, and chicken meat retailers, was conducted in a peri-urban, low to middle-income area (Dagoretti), and a very-low income informal settlement (Kibera) of Nairobi. Chicken faeces were collected using one pair of boot socks per farm, and 3 raw chicken meat samples were purchased per retailer. Samples were cultured for viable Campylobacter spp. using mCCDA, followed by blood agar plates in aerobic/microaerobic conditions for prevalence calculations. A questionnaire-based survey on sanitary, sourcing and selling practices was conducted at each site for risk factor identification using logistic regression analyses. A total of 171 farm premises and 53 retailers were sampled and interviewed. The prevalence results for Campylobacter spp. were between 33 to 44% for broiler and indigenous chicken farms, 60% and 64% for retailers, in Dagoretti and Kibera, respectively. Univariable logistic regression showed an association between Campylobacter spp. presence and the easiness of cleaning the display material used by the retailer. Restricting access to the flock was also associated with the pathogen’s presence. Multivariable logistic regression identified the selling of defrosted meat as a retailer risk factor (OR: 4.69; 95% CI: 1.31–19.97), calling for more investigation of the reported repetitive freezing-thawing processes and cold chain improvement options. At the farm-level, having a pen floor of material not easy to clean was found to increase the risk (OR: 2.31; 95%CI: 1.06–5.37). The relatively high prevalence of Campylobacter spp. across different areas and value chain nodes indicates a clear human exposure risk. The open nature of both small-scale broiler and indigenous chicken production practices with low biosecurity, hygiene and informal transactions, likely plays a role in this. While gradual improvement of farm biosecurity is recommended, risk factors identified suggest that consumer education and enforcement of basic food safety principles at the retailer end of the food continuum represent key targets for risk reduction in informal settings. [ABSTRACT FROM AUTHOR]

Mycolic acids (MA) are major, species-specific lipid components of Mycobacteria and related genera. In Mycobacterium tuberculosis, it is made up of alpha-, methoxy- and keto-MA, each with specific biological functions and conformational characteristics. Antibodies in tuberculosis (TB) patient sera respond differently towards the three MA classes and were reported to cross-react with cholesterol. To understand the antigenicity and cholesterol cross-reactivity of MA, we generated three different chicken -derived phage-displayed single-chain variable fragments (scFv) that reacted similarly towards the natural mixture of MA, but the first recognized all three classes of chemically synthetic MAs, the second only the two oxygenated types of MAs and the third only methoxy MA. The cholesterol cross-reactivity was investigated after grafting each of the three scFv types onto two configurations of constant chain domains–CH1-4 and CH2-4. Weak but significant cross-reactivity with cholesterol was found only with CH2-4 versions, notably those two that were also able to recognize the trans-keto MA. The cholesteroid nature of mycobacterial mycolic acids therefore seems to be determined by the trans-keto MA subclass. The significantly weaker binding to cholesterol in comparison to MA confirms the potential TB diagnostic application of these antibodies. [ABSTRACT FROM AUTHOR]

The aim of the study was to determine whether the use of Mn nanoparticles would make it possible to reduce the level of this micronutrient added to turkey diets without adversely affecting growth performance, antioxidant and immune status, or apoptosis. The experiment was conducted on 6 groups of turkeys with 10 replications in a two-factor design with 3 dosages of manganese, 100, 50 and 10 mg/kg, and 2 sources, manganese oxide (MnO) and manganese nanoparticles (NP-Mn2O3). The study showed that irrespective of the form of Mn used, reducing the Mn level recommended by British United Turkeys for supplementation of the diet of young turkeys from 100 mg/kg to 10 mg/kg increases the content of this element in the blood with no adverse effect on growth performance or the immune system. The reduction in Mn supplementation in the form of NP-Mn2O3 from 100 to 50 and even to 10 mg/kg of turkey diet has no negative effect on antioxidant defence in young turkeys. A 50% reduction of the recommended Mn level in the form of MnO enhances lipid oxidation processes. Replacing MnO with NP-Mn2O3 in the turkey diet probably can increase apoptosis in young turkeys. On the other hand, irrespective of the form of Mn used, reducing supplementation of the turkey diet with this element from 100 to 50 and even to 10 mg/kg probably can reduce apoptosis. [ABSTRACT FROM AUTHOR]

The diseases caused by Salmonella Gallinarum and S. Pullorum in chickens known as fowl typhoid and pullorum disease, respectively, pose a great threat to the poultry industry mainly in developing countries, since they have already been controlled in the developed ones. These bacteria are very similar at the genomic level but develop distinct host-pathogen relationships with chickens. Therefore, a deep understanding of the molecular mechanisms whereby S. Gallinarum and S. Pullorum interact with the host could lead to the development of new approaches to control and, perhaps, eradicate both diseases from the chicken flocks worldwide. Based on our previous study, it was hypothesised that metabolism-related pseudogenes, fixed in S. Pullorum genomes, could play a role in the distinct host-pathogen interaction with susceptible chickens. To test this idea, three genes (idnT, idnO and ccmH) of S. Gallinarum str. 287/91, which are pseudogenes on the S. Pullorum chromosomes, were inactivated by mutations. These genetically engineered strains grew well on the solid media without any colony morphology difference. In addition, similar growth curves were obtained by cultivation in M9 minimal medium containing D-gluconate as the sole carbon source. Infection of chickens with idnTO mutants led to increased numbers of bacteria in the livers and spleens at 5 days post-infection, but with slightly decreased heterophil infiltration in the spleens when compared to the wild-type strain. On the other hand, no significant phenotypic change was caused by mutation to ccmH genes. Apart from the above-mentioned alterations, all S. Gallinarum strains provoked similar infections, since mortality, clinical signs, macroscopic alterations and immune response were similar to the infected chickens. Therefore, according to the model applied to this study, mutation to the idnTO and ccmH genes showed minor impact on the fowl typhoid pathogenesis and so they may be relics from the ancestor genome. Our data hints at a more complex mechanism driving the distinct host-pathogen interaction of S. Gallinarum/Pullorum with chickens than differential inactivation of a few genes. [ABSTRACT FROM AUTHOR]

Fosfomycin is a broad-spectrum antibiotic with the activity against both Gram-positive and Gram-negative pathogens by inhibiting the bacterial cell wall synthesis. Given the potential therapeutic efficacy of fosfomycin against Campylobacter spp., the aim of the present study was to determine the in vitro fosfomycin susceptibility of Campylobacter isolates from avian sources including poultry and wild birds. A total of eight (1.8%) strains of Campylobacter including five C. jejuni strains isolated from ducks and three C. coli strains isolated from chickens and duck showed resistance to fosfomycin, with MICs ranging from 64 to ≥ 256 μg/mL. The extent of fosfomycin resistance was 0%, 0.9% and 3.9% in wild birds, chicken and ducks respectively. The MIC50, MIC90, and MIC100 values were 8, 32, and 32 μg/mL respectively in wild bird, 32, 32, and 64 μg/mL respectively in chicken, and 32, 32, and ≥ 256 μg/mL respectively in ducks. All eight fosfomycin-resistant Campylobacter strains were multidrug resistant; six were also resistant to fluoroquinolones, ampicillin, and tetracycline, and two were also resistant to fluoroquinolones, ampicillin, tetracycline, and macrolides. However, the fosfomycin resistance gene fosXCC was not detected in the eight fosfomycin-resistant strains. Because food animals can harbor fosfomycin-resistant Campylobacter and transmit them to humans, greater efforts are needed to monitor the prevalence of fosfomycin resistance in Campylobacter strains isolated from such animals. [ABSTRACT FROM AUTHOR]

Intestinal epithelial cells are multi-tasked cells that participate in digestion and absorption as well as in protection of the digestive tract. While information on the physiology and immune functions of intestinal epithelial cells in mammals is abundant, little is known of their immune function in birds and other species. Our main objectives were to study the development of anti-bacterial innate immune functions in the rapidly developing gut of the pre- and post-hatch chick and to determine the functional diversity of epithelial cells. After establishing primary intestinal epithelial cell cultures, we demonstrated their capacity to uptake and process bacteria. The response to bacterial products, LPS and LTA, induced expression of pro-inflammatory cytokine genes (IL-6, IL-18) as well as the expression of the acute phase proteins avidin, lysozyme and the secretory component derived from the polymeric immunoglobulin receptor. These proteins were then localized in gut sections, and the goblet cell was shown to store avidin, lysozyme as well as secretory component. Lysozyme staining was also located in a novel rod-shaped intestinal cell, situated at different loci along the villus, thus deviating from the classical Paneth cell in the mammal, that is restricted to crypts. Thus, in the chicken, the intestinal epithelium, and particularly goblet cells, are committed to innate immune protection. The unique role of the goblet cell in chicken intestinal immunity, as well as the unique distribution of lysozyme-positive cells highlight alternative solutions of gut protection in the bird. [ABSTRACT FROM AUTHOR]

Several antimicrobials are routinely used by the poultry farming industry on their daily operations, however, researchers have found for some antimicrobials that their residues persist for longer periods in feathers than they do in edible tissues, and at higher concentrations, as well. But this information is not known for other classes of antimicrobials, such as the sulfonamides. Therefore, this work presents an accurate and reliable analytical method for the detection of sulfachloropyridazine (SCP) in feathers and edible tissues from broiler chickens. This method was also validated in-house and then used to study the depletion of sulfachloropyridazine in those matrices. The experimental group comprised 54 broiler chickens, who were raised under controlled conditions and then treated with a commercial formulation of 10% sulfachloropyridazine for 5 days. Samples were analyzed via LC-MS/MS, using 13C6-sulfamethazine (SMZ-13C6) as an internal standard. Aromatic sulfonic acid solid phase extraction (SPE) cartridges were used to clean up the samples. The Limit of Detection (LOD) for this method was set at 10 μg kg-1 on feathers and liver; and at 5 μg kg-1 on muscle. Within the range of 10–100 μg kg-1, the calibration curves for all matrices presented a determination coefficient greater than 0.96. Our results show, with a 95% confidence level, that sulfachloropyridazine persisted in feathers for up to 55 days after ceasing treatment, and its concentrations were higher than in edible tissues. In consequence, to avoid re-entry of antimicrobial residues into the food-chain, we recommend monitoring and inspecting animal diets that contain feather derivatives, such as feathers meals, because they could be sourced from birds that might have been medicated with sulfachloropyridazine. [ABSTRACT FROM AUTHOR]

The genotypes of the H9N2 avian influenza viruses have changed since 2013 when almost all H9N2 viruses circulating in chickens in China were genotype 57 (G57) with the fittest lineage of each gene. To characterize the H9N2 variant viruses from 2011 to 2014, 28 H9N2 influenza viruses were isolated from live poultry markets in China from 2011–2014 and were analyzed by genetic and biological characterization. Our findings showed that 16 residues that changed antigenicity, two potential N-linked glycosylation sites, and one amino acid in the receptor binding site of the HA protein changed significantly from 2011–2014. Moreover, the HA and NA genes in the phylogenetic tree were mainly clustered into two independent branches, A and B, based on the year of isolation. H9N2 virus internal genes were related to those from the human-infected avian influenza viruses H5N1, H7N9, and H10N8. In particular, the NS gene in the phylogenetic tree revealed genetic divergence of the virus gene into three branches labeled A, B, and C, which were related to the H9N2, H10N8, and H7N9 viruses, respectively. Additionally, the isolates also showed varying levels of infection and airborne transmission. These results indicated that the H9N2 virus had undergone an adaptive evolution and variation from 2011–2014. [ABSTRACT FROM AUTHOR]

This paper focuses on a novel methodology of subjective speech quality measurement and repeatability of its results between laboratory conditions and simulated environmental conditions. A single set of speech samples was distorted by various background noises and low bit-rate coding techniques. This study aimed to compare results of subjective speech quality tests with and without a parallel task deploying the ITU-T P.835 methodology. Afterward, tests results performed with and without a parallel task were compared using Pearson correlation, CI95, and numbers of opposite pair-wise comparisons. The tests show differences in results in the case of a parallel task. [ABSTRACT FROM AUTHOR]

Allogeneic lung transplant is limited both by the shortage of available donor lungs and by the lack of suitable long-term lung assist devices to bridge patients to lung transplantation. Avian lungs have different structure and mechanics resulting in more efficient gas exchange than mammalian lungs. Decellularized avian lungs, recellularized with human lung cells, could therefore provide a powerful novel gas exchange unit for potential use in pulmonary therapeutics. To initially assess this in both small and large avian lung models, chicken (Gallus gallus domesticus) and emu (Dromaius novaehollandiae) lungs were decellularized using modifications of a detergent-based protocol, previously utilized with mammalian lungs. Light and electron microscopy, vascular and airway resistance, quantitation and gel analyses of residual DNA, and immunohistochemical and mass spectrometric analyses of remaining extracellular matrix (ECM) proteins demonstrated maintenance of lung structure, minimal residual DNA, and retention of major ECM proteins in the decellularized scaffolds. Seeding with human bronchial epithelial cells, human pulmonary vascular endothelial cells, human mesenchymal stromal cells, and human lung fibroblasts demonstrated initial cell attachment on decellularized avian lungs and growth over a 7-day period. These initial studies demonstrate that decellularized avian lungs may be a feasible approach for generating functional lung tissue for clinical therapeutics. [ABSTRACT FROM AUTHOR]

The alternative prey hypothesis predicts that the interaction between generalist predators and their main prey is a major driver of population dynamics of alternative prey species. In Fennoscandia, changes in climate and human land use are assumed to alter the dynamics of cyclic small rodents (main prey) and lead to increased densities and range expansion of an important generalist predator, the red fox Vulpes vulpes. In order to better understand the role of these potential changes in community structure on an alternative prey species, willow ptarmigan Lagopus lagopus, we analyzed nine years of population census data from SE Norway to investigate how community interactions affected their population dynamics. The ptarmigan populations showed no declining trend during the study period, and annual variations corresponded with marked periodic small rodent peaks and declines. Population growth and breeding success were highly correlated, and both demographic variables were influenced by an interaction between red fox and small rodents. Red foxes affected ptarmigan negatively only when small rodent abundance was low, which is in accordance with the alternative prey hypothesis. Our results confirm the important role of red fox predation in ptarmigan dynamics, and indicate that if small rodent cycles are disrupted, this may lead to decline in ptarmigan and other alternative prey species due to elevated predation pressure. [ABSTRACT FROM AUTHOR]

Host Defense Peptides (HDPs) such as cathelicidins are multifunctional effectors of the innate immune system with both antimicrobial and pleiotropic immunomodulatory functions. Chicken cathelicidin-2 (CATH-2) has multiple immunomodulatory effects in vitro and the D-amino acid analog of this peptide has been shown to partially protect young chicks from a bacterial infection. However, the mechanisms responsible for CATH-2 mediated in vivo protection have not been investigated so far. In this study, D-CATH-2 was administered in ovo and the immune status and microbiota of the chicks were investigated at 7 days posthatch to elucidate the in vivo mechanisms of the peptide. In three consecutive studies, no effects on numbers and functions of immune cells were found and only small changes were seen in gene expression of Peripheral Blood Mononuclear Cells (PBMCs). In two studies, intestinal microbiota composition was determined which was highly variable, suggesting that it was strongly influenced by environmental factors. In both studies, in ovo D-CATH-2 treatment caused significant reduction of Ruminococcaceae and Butyricicoccus in the cecum and Escherichia/Shigella in both ileum and cecum. In conclusion, this study shows that, in the absence of an infectious stimulus, in ovo administration of a CATH-2 analog alters the microbiota composition but does not affect the chicks’ immune system posthatch. [ABSTRACT FROM AUTHOR]

A mixed-methods approach was used to evaluate and improve the “late-night dining” options in a university dining hall. Surveys assessed student desires for late-night offerings, and evaluated students’ habits and motivations during late-night dining. Two interventions were implemented to see if students could be “nudged” into different choice patterns. In the first, a “veggie-heavy” entrée was added at the beginning of the entrée line, so that students would substitute an entrée containing vegetables for the alternatives. In the second, a snack-food bar was set up to cater to students who didn’t want to stand in the long entrée line, and preferred a snack. Data on food choice was collected during the interventions. Survey responses showed significant differences in the reasons females and males utilized late-night dining (p<0.001). We also found that students at late-night dining had a lower emphasis on health than the general student population. Even students at late-night who reported being health-conscious showed no difference in food selections from students who said health was not important (p = 0.883). Veggie-heavy entrées had mild success in increasing vegetable selection. However, veggie-heavy entrées were largely ignored when the other option was chicken nuggets. The snack bar was very popular. Entrée placement and convenience lines may have mild impacts on food selection in a late-night dining environment. [ABSTRACT FROM AUTHOR]

The transgenic chicken has been considered as a prospective bioreactor for large-scale production of costly pharmaceutical proteins. In the present study, we report successful generation of transgenic hens that lay eggs containing a high concentration of human erythropoietin (hEPO) in the ovalbumin. Using a feline immunodeficiency virus (FIV)-based pseudotyped lentivirus vector enveloped with G glycoproteins of the vesicular stomatitis virus, the replication-defective vector virus carrying the hEPO gene under the control of the chicken ovalbumin promoter was microinjected to the subgerminal cavity of freshly laid chicken eggs (stage X). Stable germline transmission of the hEPO transgene to the G1 progeny, which were non-mosaic and hemizygous for the hEPO gene under the ovalbumin promoter, was confirmed by mating of a G0 rooster with non-transgenic hens. Quantitative analysis of hEPO in the egg whites and in the blood samples taken from G1 transgenic chickens showed 4,810 ~ 6,600 IU/ml (40.1 ~ 55.0 μg/ml) and almost no detectable concentration, respectively, indicating tightly regulated oviduct-specific expression of the hEPO transgene. In terms of biological activity, there was no difference between the recombinant hEPO contained in the transgenic egg white and the commercially available counterpart, in vitro. We suggest that these results imply an important step toward efficient production of human cytokines from a transgenic animal bioreactor. [ABSTRACT FROM AUTHOR]

Iron is one of the most important elements for bacterial survival and pathogenicity. The iron uptake mechanism of Riemerella anatipestifer (R. anatipestifer, RA), a major pathogen that causes septicemia and polyserositis in ducks, is largely unknown. Here, the functions of the putative TonB-dependent iron transporter of RA-CH-1, B739_1343, in iron utilization and pathogenicity were investigated. Under iron-starved conditions, the mutant strain RA-CH-1ΔB739_1343 exhibited more seriously impaired growth than the wild-type strain RA-CH-1, and the expression of B739_1343 in the mutant strain restored growth. qRT-PCR results showed that the transcription of B739_1343 was not regulated by iron conditions. In an animal model, the median lethal dose (LD50) of the mutant strain RA-CH-1ΔB739_1343 increased more than 104-fold (1.6×1012 CFU) compared to that of the wild-type strain RA-CH-1 (1.43×108 CFU). In a duck co-infection model, the mutant strain RA-CH-1ΔB739_1343 was outcompeted by the wild-type RA-CH-1 in the blood, liver and brain of infected ducks, indicating that B739_1343 is a virulence factor of RA-CH-1. Finally, immunization with live bacteria of the mutant strain RA-CH-1ΔB739_1343 protected 83.33% of ducks against a high-dose (100-fold LD50) challenge with the wild-type strain RA-CH-1, suggesting that the mutant strain RA-CH-1ΔB739_1343 could be further developed as a potential live attenuated vaccine candidate for the duck industry. [ABSTRACT FROM AUTHOR]

Borrelia burgdorferi sensu lato (Bbsl), the causative agent of Lyme disease, establishes an initial infection in the host’s skin following a tick bite, and then disseminates to distant organs, leading to multisystem manifestations. Tick-to-vertebrate host transmission requires that Bbsl survives during blood feeding. Complement is an important innate host defense in blood and interstitial fluid. Bbsl produces a polymorphic surface protein, CspA, that binds to a complement regulator, Factor H (FH) to block complement activation in vitro. However, the role that CspA plays in the Bbsl enzootic cycle remains unclear. In this study, we demonstrated that different CspA variants promote spirochete binding to FH to inactivate complement and promote serum resistance in a host-specific manner. Utilizing a tick-to-mouse transmission model, we observed that a cspA-knockout B. burgdorferi is eliminated from nymphal ticks in the first 24 hours of feeding and is unable to be transmitted to naïve mice. Conversely, ectopically producing CspA derived from B. burgdorferi or B. afzelii, but not B. garinii in a cspA-knockout strain restored spirochete survival in fed nymphs and tick-to-mouse transmission. Furthermore, a CspA point mutant, CspA-L246D that was defective in FH-binding, failed to survive in fed nymphs and at the inoculation site or bloodstream in mice. We also allowed those spirochete-infected nymphs to feed on C3-/- mice that lacked functional complement. The cspA-knockout B. burgdorferi or this mutant strain complemented with cspA variants or cspA-L246D was found at similar levels as wild type B. burgdorferi in the fed nymphs and mouse tissues. These novel findings suggest that the FH-binding activity of CspA protects spirochetes from complement-mediated killing in fed nymphal ticks, which ultimately allows Bbsl transmission to mammalian hosts. [ABSTRACT FROM AUTHOR]

Meat quality is closely related to adipose tissues in ducks, and adipogenesis is controlled by a complex network of transcription factors tightly acting at different stages of differentiation especially in ducks. The aim of this study was to establish the preadipocyte in vitro culture system and understand the biological characteristics of expansion of duck adipocyte tissue at the cellular and molecular level. We isolated pre-adipocytes from the subcutaneous fat of three breeds of duck and differentiated them into mature adipocytes using a mixture of insulin, rosiglitazone, dexamethasone, 3-isobutyl-1-methylxanthine, and oleic acid over 0,2, 4, 6, and 8 days. Successful differentiation was confirmed from the development of lipid droplets and their response to Oil Red O, and increasing numbers of lipid droplets were stained red over time. The expression of key marker genes, including peroxisome proliferator activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα), adipocyte fatty acid binding protein 4 (FABP4), and fatty acid synthetase (FAS), gradually increased during pre-adipocyte differentiation. Furthermore, it was verified by interference experiments that the knockdown of PPARγ directly reduced lipid production. Meanwhile we analyzed the role of unsaturated fatty acids in the production of poultry fat using different concentrations of oleic acid and found that lipid droplet deposition was highest when the concentration of oleic acid was 300 μM. We also compared the level of differentiated pre-adipocytes that were isolated from Jianchang ducks (fatty-meat duck), Cherry Valley ducks (lean-meat duck) and White-crested ducks (egg-producing duck). The proliferation and differentiation rate of pre-adipocytes derived from Jianchang ducks was higher than that of White-crested ducks. These results provide the foundation for further research into waterfowl adipogenesis. [ABSTRACT FROM AUTHOR]

Newcastle disease (ND) causes severe economic loss to poultry industry worldwide. Frequent outbreaks of ND in commercial chickens vaccinated with live vaccines suggest a need to develop improved vaccines that are genetically matched against circulating Newcastle disease virus (NDV) strains. In this study, the fusion protein cleavage site (FPCS) sequence of NDV strain Banjarmasin/010 (Banj), a genotype VII NDV, was individually modified using primer mutagenesis to those of avian paramyxovirus (APMV) serotypes 2, 7 and 8 and compared with the recombinant Banjarmasin (rBanj) with avirulent NDV LaSota cleavage site (rBanj-LaSota). These FPCS mutations changed the in vitro cell-to-cell fusion activity and made rBanj FPCS mutant viruses highly attenuated in chickens. When chickens immunized with the rBanj FPCS mutant viruses and challenged with the virulent Banj, there was reduced challenge virus shedding observed compared to chickens immunized with the heterologous vaccine strain LaSota. Among the genotype VII NDV Banj vaccine candidates, rBanj-LaSota and rBanj containing FPCS of APMV-8 induced highest neutralizing antibody titers and protected chickens with reduced challenge virus shedding. These results show the effect of the F protein cleavage site sequence in generating genotype VII matched NDV vaccines. [ABSTRACT FROM AUTHOR]