Screening of polysaccharides from the differently processed products of Angelica sinensis with the best liver protection effect on chicken and the intervention mechanism study based on tandem mass tag proteomics and multiple reaction monitoring approach.

The incidence of colibacillosis in poultry is on the rise, significantly affecting the chicken industry. Ceftiofur sodium (CS) is frequently employed to treat this disease, resulting in lipopolysaccharide (LPS) buildup. Processing plays a vital role in traditional Chinese veterinary medicine. The potential intervention in liver injury by polysaccharides from the differently processed products of Angelica sinensis (PDPPAS) induced by combined CS and LPS remains unclear. This study aims to investigate the protective effect of PDPPAS on chicken liver injury caused by CS combined with LPS buildup and further identify the polysaccharides with the highest hepatoprotective activity in chickens. Furthermore, the study elucidates polysaccharides' intervention mechanism using tandem mass tag (TMT) proteomics and multiple reaction monitoring (MRM) methods. A total of 190 1-day-old layer chickens were randomly assigned into 12 groups, of which 14 chickens were in the control group and 16 in other groups, for a 10-day trial. The screening results showed that charred A. sinensis polysaccharide (CASP) had the most effective and the best hepatoprotective effect at 48 h. TMT proteomics and MRM validation results demonstrated that the intervention mechanism of the CASP high-dose (CASPH) intervention group was closely related to the protein expressions of FCER2, TBXAS1, CD34, AGXT, GCAT, COX7A2L, and CYP2AC1. Conclusively, the intervention mechanism of CASPH had multitarget, multicenter regulatory features.

Effect of papaya (Carica papaya) seed as phytogenic feed additives on egg performance, egg quality and blood serum biochemical constituents of layer hens.

BACKGROUND: Supplementing poultry diets with certain natural phytogenic additives has positive impacts on performance parameters like feed intake, egg production and quality of layer hens. OBJECTIVES: The study aimed to examine how supplementing papaya seed as phytogenic feed additive (PFA) to the diets of layer chickens affected their egg production, quality and blood serum biochemical parameters. METHODS: One hundred twenty 28-week-old Bovans brown laying hens were randomly assigned to one of four treatments with varying levels of papaya seed supplementation at 0 g/kg (T1), 5 g/kg (T2), 10 g/kg (T3) and 15 g/kg (T4) in a complete randomized design. RESULT: Layer chickens supplemented with 0.5% (T2), 1% (T3) and 1.5% (T4) papaya seed as a PFA had significantly (p < 0.05) improved dry matter intake, egg production%, egg mass and feed conversion. On the other hand, papaya seed supplementation decreased (p < 0.05) the bodyweight gain of laying hens. Papaya seed supplementation significantly (p < 0.05) increased egg weight, length, width, yolk height and yolk colour compared to the control group. Serum total protein on T4 and liver enzyme of alkaline phosphatase on T3 were significantly greater (p < 0.05) than in the control group. However, there were no significant differences (p > 0.05) in serum albumin, glucose, total cholesterol, triglyceride and aspartate aminotransferase levels among treatments laying hens. CONCLUSION: Papaya seed can be a potential candidate as a PFA at 5-15 g/kg of the basal diet to improve egg-laying performance and quality without deteriorating effect.

Segmented filamentous bacteria-based treatment to elicit protection against Enterobacteriaceae in Layer chickens.

Introduction: Gut microbes like segmented filamentous bacteria (SFB) play a key role in gut maturation during early life, as demonstrated in humans and mice. Our previous study demonstrated oral inoculation of ileum-spores containing SFB to chickens after hatch increases early SFB gut colonization, which increases immune maturation and resistance to bacteria, like Salmonella, as tested in vitro; however, more studies are needed for treatment optimization and in vivo testing. The objectives of this study were to (1) test a treatment that includes both spores and filamentous SFB, (2) validate antimicrobial ability of the treatment in layer hens in vivo, and (3) elucidate its molecular mechanism. Methods: One-day-old specific pathogen-free layers (n = 12 per group) were orally treated with either PBS (CON) or SFB-based treatment (SFB). At 4 days post-inoculation (DPI), both CON and SFB groups were orally challenged with Salmonella Typhimurium. Total Enterobacteriaceae and Salmonella were examined by plating and enumeration in feces at 7,10 and 14 dpi; and in the ileum, cecum, and spleen at 16 dpi in euthanized birds. The presence and levels of SFB were determined from ilea scrapings via microscopy and qPCR, respectively. Relative gene expression of host-derived antimicrobial peptides and cytokines in the distal ileum was determined by RT-qPCR. Results: At 10 and 14 dpi, a significant decrease in total Enterobacteriaceae was observed in the feces of the SFB group. At necropsy, the level of SFB was significantly higher in the SFB group than in the CON group, while a significant decrease in total Enterobacteriaceae and Salmonella was observed in the ceca of the SFB group. RT-qPCR revealed increased expression of ß-defensin 14, and cytokines IL-10 and IFNγ. Discussion: The introduction of SFB at hatch as a prophylactic treatment may benefit commercial partners as well as consumers by reducing the incidence of Enterobacteriaceae in food animals. Reduction of these bacteria in animals would, in turn, increase animal health, productivity, and safety for consumers. Studies to optimize the treatment for poultry industry applications are ongoing in our lab.

Estimates of on-farm antimicrobial usage in egg production in the United States, 2016-2021.

Very few data exist globally regarding the use of antimicrobials in the table egg industry. Antimicrobial use data from broiler chickens and turkeys cannot be used as a surrogate of layer chickens because of the fact that table eggs for human consumption are produced daily by laying hens. To avoid the possibility of antimicrobial residues in the eggs, there are very few antimicrobials approved for use in layers in the U.S. The objective of this study was to collect on-farm antimicrobial use data from the U.S. table egg industry and to have it be representative of the national layer flock. Participation was voluntary. Data were collected for the period 2016 through 2021 and are reported on a calendar year basis. Using production statistics from USDA:NASS as a denominator, the data supplied by participating companies accounted for 3,016,183,140 dozen eggs (~40% of national egg production) in 2016 and 3,556,743,270 dozen eggs (~45% of national egg production) in 2021. All of the replacement chicks placed on pullet farms during the study period were estimated to have received 0.2 mg/chick gentamicin at the hatchery. Most of the antimicrobial administration in U.S. egg production is via the feed. The ionophores monensin and salinomycin were used in the pullets, bacitracin was used in both pullets and layers (primarily for control of necrotic enteritis), and chlortetracycline was used primarily in layers for the treatment of E. coli-related disease. In the layers, between 0.10 and 0.19% of total hen-days were exposed to chlortetracycline. Only two water-soluble administrations were recorded during the entire study period, both involving lincomycin to pullet flocks for the treatment of necrotic enteritis. Overall, antimicrobial use in the U.S. layer industry was focused mainly on controlling necrotic enteritis in the pullets and treating E. coli-related disease in the laying hens.

Study on the Hepatoprotective Effect Mechanism of Polysaccharides from Charred Angelica sinensis on the Layer Chickens Based on the Detection of the Intestinal Floras and Short-Chain Fatty Acids of Cecal Contents and Association Analysis.

To analyze the intervention mechanism of polysaccharides from charred Angelica sinensis (CASP) on the liver injury caused by Ceftiofur sodium (CS) and lipopolysaccharide (LPS) from the perspective of the intestine. Ninety-four one-day-old laying chickens underwent free feeding and drinking water for three days. Then, fourteen laying chickens were randomly selected as the control group, and sixteen laying chickens were selected as the model group. Sixteen laying chickens in the rest were randomly selected as the intervention group of CASP. Chickens in the intervention group were given CASP by the oral administration (0.25 g/kg/d) for 10 days, the control and model groups were given the same amount of physiological saline. During the 8th and 10th days, laying chickens in the model and CASP intervention group were subcutaneously injected with CS at the neck. In contrast, those in the control group were subcutaneously injected with the same amount of normal saline simultaneously. Except for the control group, the layer chickens in the model and CASP intervention groups were injected with LPS after CS injection on the 10th day of the experiment. In contrast, those in the control group were injected with the same amount of normal saline at the same time. 48 h after the experiment, the liver samples of each group were collected, and the liver injury was analyzed by hematoxylin-eosin (HE) staining and transmission electron microscopy. And the cecum contents of six-layer chickens in each group were collected, and the intervention mechanism of CASP on the liver injury from the perspective of the intestine was analyzed by the 16S rDNA amplicon sequencing technology and the short-chain fatty acids (SCFAs) detection of cecal contents based on Gas Chromatography-Mass Spectrometry (GC-MS), and their association analysis was carried out. The results showed that the structure of chicken liver in the normal control group was normal, while that in the model group was damaged. The structure of chicken liver in the CASP intervention group was similar to the normal control group. The intestinal floras in the model group were maladjusted compared to the normal control group. After the intervention of CASP, the diversity, and richness of chicken intestinal floras changed significantly. It was speculated that the intervention mechanism of CASP on the chicken liver injury might be related to the abundance and proportion of Bacteroidetes and Firmicutes. Compared with the model group, the indexes of ace, chao1, observed species, and PD whole tree of chicken cecum floras in the intervention group of CASP were significantly increased (p < 0.05). The contents of acetic acid, butyric acid, and total SCFAs in the intervention group of CASP were significantly lower than those in the model group (p < 0.05), and the contents of propionic acid and valeric acid in the intervention group of CASP were significantly lower than those in the model group (p < 0.05) and normal control group (p < 0.05). The correlation analysis showed that the changes in the intestinal floras were correlated with the changes in SCFAs in the cecum. It is confirmed that the liver-protecting effect of CASP is indeed related to the changes in the intestinal floras and SCFAs content in the cecum, which provides a basis for screening liver-protecting alternative antibiotics products for poultry.

Antimicrobial resistance in bacteria isolated from the poultry production system in Nepal.

SETTING: Twenty poultry farms in five provinces of Nepal were selected for studying bacterial pathogens and their antimicrobial resistance (AMR) patterns. OBJECTIVE: To document the proportion of cloacal swabs collected from 3,230 broiler and 3,230 layer chickens from September to December 2021 that grew isolates of Escherichia coli, Enterococcus spp. and Salmonella spp. along with their AMR patterns. DESIGN: This was a cross-sectional descriptive study. RESULTS: In broiler birds, Enterococcus spp., Salmonella spp. and E. coli were identified in respectively 36%, 39% and 63% of swabs. In layer birds, Enterococcus spp., Salmonella spp. and E. coli were identified in respectively 31%, 48% and 60% of swabs. For both bird types, there was variation in bacterial prevalence between the regions. For all three bacterial isolates, the lowest antimicrobial resistance was found with amikacin. For the other nine antibiotics tested, >50% of bacterial isolates showed resistance; between 60% and 90% of isolates showed resistance to ciprofloxacin and trimethoprim-sulfamethoxazole. Multidrug resistance ranged from 45% to 46% for Salmonella spp., 37-44% for E. coli and 13-17% for Enterococcus spp. CONCLUSION: This study shows that a large proportion of poultry in Nepal are infected with potentially pathogenic bacteria, and these are frequently resistant to commonly used antibiotics. Nepal urgently needs to implement corrective measures.

CONTEXTE: Vingt fermes avicoles dans cinq provinces du Népal ont été sélectionnées pour étudier les pathogènes bactériens et leurs profils de résistance aux antimicrobiens (AMR). OBJECTIF: Documenter la proportion d'écouvillons cloacaux prélevés chez 3 230 poulets de chair et 3 230 poules pondeuses de septembre à décembre 2021 qui ont produit des isolats d'Escherichia coli, d'Enterococcus spp. et de Salmonella spp. ainsi que leurs profils d'AMR. MÉTHODE: Il s'agissait d'une étude descriptive transversale. RÉSULTATS: Chez les poulets de chair, Enterococcus spp., Salmonella spp. et E. coli ont été identifiés dans respectivement 36%, 39% et 63% des écouvillons. Chez les pondeuses, Enterococcus spp., Salmonella spp. et E. coli ont été identifiés dans respectivement 31%, 48% et 60% des écouvillons. Pour les deux types d'oiseaux, la prévalence bactérienne varie selon les régions. Pour les trois isolats bactériens, la résistance la plus faible a été observée avec l'amikacine. Pour les neuf autres antibiotiques testés, >50% des isolats bactériens présentaient une résistance ; entre 60% et 90% des isolats présentaient une résistance à la ciprofloxacine et au triméthoprime-sulfaméthoxazole. La multirésistance variait de 45 à 46% pour Salmonella spp, 37 à 44% pour E. coli et 13 à 17% pour Enterococcus spp. CONCLUSION: Cette étude montre qu'une grande proportion de volailles au Népal est infectée par des bactéries potentiellement pathogènes, et que celles-ci sont fréquemment résistantes aux antibiotiques couramment utilisés. Le Népal doit de toute urgence mettre en œuvre des mesures correctives.

Effects of Dietary Inclusion of ß-Hydroxy-ß-Methylbutyrate on Growth Performance, Fat Deposition, Bile Acid Metabolism, and Gut Microbiota Function in High-Fat and High-Cholesterol Diet-Challenged Layer Chickens.

Excessive lipid deposition in layer chickens due to inappropriate feeding adversely affects egg production; however, nutritional manipulation methods to deal with this issue are still limited. ß-hydroxy-ß-methylbutyrate (HMB), a metabolite of L-leucine, was recently reported as a lipid-lowering nutrient in mice and pigs, although its role in layers had not been investigated. Here, we employed high-fat and high-cholesterol diet (HFHCD)-challenged growing layers as an obese model to explore HMB function in the regulation of lipid metabolism and the potential mechanisms involved. We found that dietary supplementation with (0.05% or 0.10%) HMB significantly reduced HFHCD-induced bodyweight growth in layers, mainly due to reduction in abdominal fat deposition. Mechanistically, HMB supplementation enhanced hepatic bile acid synthesis from cholesterol through elevating expression of Cyp7a1, a gene coding a key enzyme in bile acid synthesis. Furthermore, 16S rRNA gene sequencing revealed that HMB supplementation remodeled the diversity and composition of the layers' cecal microbiota, and the abundance of Bacteroidetes at the phylum level were especially affected. Correlation analysis further indicated a strong negative association between Bacteroidetes abundance and lipid metabolism-related parameters. Taken together, these data suggest that dietary HMB supplementation could improve abdominal fat deposition in layers, probably through modulating hepatic bile acid synthesis and gut microbiota function.

Effects of blue-green LED lights with two perceived illuminance (human and poultry) on immune performance and skeletal development of layer chickens.

Light is one of the essential environmental factors in the production process of laying hens, which can directly affect their behavior, growth and development, and production performance. The spectral sensitivity of humans is different from that of poultry, and the perceived illuminance units of human and poultry are lux and clux, respectively. If the light management of laying hen production is carried out according to human perceived illuminance, the growth and development of laying hens during pullet rearing may be adversely affected due to the discomfort of the perceived illuminance. Preliminary research has found that blue-green LED light can improve the immune function of laying hens during the brooding and rearing periods. However, the differences of the effects caused by blue-green light on the immune performance and bone development of laying hens during pullet rearing are still unclear for the 2 spectral sensitivities. A total of 120 Jinghong layer chickens were raised from 1 d to 13 wk of age in one of three groups with a white LED light (light intensity unit lux, WL) group, a blue-green LED light (light intensity unit lux, HBGL) group, and blue-green LED light (light intensity unit clux, PBGL) group, and unlimited feed and water were provided during the whole experiment. At 7 and 13 wk of age, the immune performance, bone parameters, and related gene expression were investigated. The results showed that compared with the WL groups, HBGL and PBGL increased the immunoglobulin A (IgA) content at 13 wk of age and the IgM content at 7 wk of age (P < 0.05). The bone mineral density (BMD) at 7 and 13 wk of age and tibial strength (TS) at 13 wk of age of the pullets in the WL group were significantly higher than those in the HBGL and PBGL group (P < 0.05). Osteoclastogenesis inhibitory factor gene (OPG mRNA) expression was increased in the layer chickens at the age of 7 and 13 wk for the WL group (P < 0.05). Compared with the WL group and PBGL group, the melanopsin gene (OPN4 mRNA) transcription level of hypothalamus and pineal gland of the chickens under HBGL significantly increased at 7 and 13 wk of age (P < 0.05). In conclusion, blue-green LED light with two perceived illuminance (human and poultry) can increase the Ig content and the immune performance of layer chickens, and blue-green LED light (light intensity unit lux) can promote the expression of OPN4 gene in the hypothalamus and pineal gland. In addition, white LED light can enhance bone quality by increasing tibia OPG gene expression.

Metabonomics study on the hepatoprotective effect mechanism of polysaccharides from different processed products of Angelica sinensis on layer chickens based on UPLC-Q/TOF-MS/MS, multivariate statistical analysis and conjoint analysis.

Chicken colibacillosis is one of the most severe diseases in the poultry industry. Ceftiofur sodium (CS) is often used to treat it in clinical practice and lipopolysaccharide (LPS) accumulates in the chicken's body. Previous experimental studies found that CS combined with LPS could induce liver injury in layer chickens, and polysaccharides from charred Angelica sinensis(CASP) had a better hepatoprotective effect than polysaccharides from unprocessed Angelica sinensis(UASP). However, the intervention mechanism was unclear. Thus, UPLC-Q/TOF-MS/MS-based metabonomics and transcriptomics were used in this study to clarify the hepatoprotective effect mechanism of CASP and UASP in layer chickens. Transcriptomics and enzyme-linked immunosorbent assay were used for biological verification of some critical mutual metabolic pathways screened with metabonomics. The comprehensive analysis results showed that in a layer chicken liver injury model built with LPS and CS, 12 critical metabolic pathways were disturbed, involving 10 important differential metabolites. The hepatoprotective effect mechanism of CASP is related to the arachidonic acid metabolism and mTOR signaling pathways, involving nine important differential metabolites. In contrast, the hepatoprotective effect mechanism of UASP is related to the arachidonic acid metabolism pathway, involving six important differential metabolites.

Surveillance on respiratory diseases reveals enzootic circulation of both H5 and H9 avian influenza viruses in small-scale commercial layer farms of Bangladesh.

Poultry production in Bangladesh has been experiencing H5N1 highly pathogenic avian influenza (HPAI) and H9N2 low pathogenic avian influenza (LPAI) for the last 14 years. Vaccination of chickens against H5 HPAI is in practice since the end of 2012. Subsequently, the official reporting of HPAI outbreaks gradually decreased. However, the true extent of circulation of avian influenza virus (AIV) in commercial poultry production is not clear. To explore this, we conducted active surveillance in 422 small-scale commercial layer farms in 20 villages of Mymensingh and Tangail districts of Bangladesh during 2017 and 2018 for the presence of diseases with respiratory signs. A total of 88 farms with respiratory disease problems were identified and investigated during the surveillance. In addition, 22 small-scale commercial layer farms in the neighbouring areas with respiratory disease problem were also investigated on request from the farmers. Pooled samples of oropharyngeal swabs from live birds or respiratory tissues from dead birds of the farm suffering from respiratory disease problem were tested for molecular detection of avian influenza virus (AIV), Newcastle disease virus (NDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum and Avibacterium paragallinarum. A total of 110 farms (88 in the surveillance site and 22 in the neighbouring region) were investigated, and one or more respiratory pathogens were detected from 89 farms. AIV was detected in 57 farms often concurrently with other pathogens. Among these 57 farms, H5, H9, both H5 and H9 or non-H5 and non-H9 AIV were detected in 28, 9, 13 or 7 farms, respectively. Birds of most of the H5 AIV-positive farms did not present typical clinical signs or high mortality. Twenty such farms were observed longitudinally, which had only 1.05%-5.50% mortality but a marked drop in egg production. This widespread circulation of H5 AIV along with H9 AIV and other pathogens in small-scale commercial layer farms, often with low mortality, reaffirms the enzootic circulation of AIV in Bangladesh, which may escape syndromic surveillance focused on unusual mortality only. To reduce public health risks, strengthening of the control programme with comprehensive vaccination, enhanced biosecurity, improved surveillance and outbreak response is suggested.

Identification of a Putative Novel Genotype of Avian Hepatitis E Virus from Apparently Healthy Chickens in Southwestern Nigeria.

Avian hepatitis E virus (aHEV) is the major etiological agent of hepatitis-splenomegaly syndrome (HSS), big liver and spleen disease (BLSD), and hepatic rupture hemorrhage syndrome (HRHS) in chickens. Infections with aHEV cause a significant decrease in egg production and increased mortality in chickens worldwide. However, studies on the prevalence of aHEV in Nigeria are scarce. In this study, serum (n = 88) and fecal samples (n = 110) obtained from apparently healthy layer chickens from three states in southwestern Nigeria were analyzed by nested reverse transcription-polymerase chain reaction (nRT-PCR) targeting the helicase and capsid gene for the presence of aHEV. Avian HEV was detected in 12.5% (n = 11/88) of serum samples and 9.1% (n = 10/110) of fecal samples tested. Phylogenetic analysis showed that five of the twelve identified aHEV sequences belonged to genotype 2. The remaining seven sequences were only distantly related to other known aHEV isolates. After amplification of the near-complete ORF2 fragment (1618 bp) and part of the ORF1 (582 bp) of isolate YF40_aHEV_NG phylogenetic analysis revealed a nucleotide sequence identity between 79.0 and 82.6% and 80.1 and 83.5%, respectively, to other known aHEV strains, indicating that the Nigerian isolate YF40_aHEV_NG belongs to a novel aHEV genotype. This is the first report of co-circulation of aHEV genotypes in chickens in Nigeria.

Modulation of folliculogenesis in adult laying chickens by bisphenol A and bisphenol S: Perspectives on ovarian morphology and gene expression.

Both bisphenol A (BPA) and its analog bisphenol S (BPS) are industrial chemicals that have been used to make certain plastic products applied in chicken farms, including food and water containers. They are endocrine disrupting chemicals (EDCs) with xenoestrogenic activities and affect reproductive success in many ways. It was hypothesized that BPA and BPS could adversely affect the folliculogenesis in chickens due to their disruption of the estrogen responses, using either genomic or non-genomic mechanisms. This study investigated the deleterious effects of BPA and BPS on the ovaries when adult layer chickens were orally treated with these EDCs at 50 µg/kg body weight, the reference dose for chronic oral exposure of BPA established by the U.S. EPA. The chickens in both BPA and BPS-treated groups showed a decreased number of the preovulatory follicles. BPA-treated chickens showed a significant decrease in the diameter of F1. Additionally, both BPA and BPS treatments increased the infiltrations of lymphocytes and plasma cells in ovaries. Moreover, it was found that the ovaries of BPS-treated chickens weighed the most among the groups. RNA sequencing and subsequent pathway enrichment analysis of differentially expressed genes revealed that both BPA- and BPS-treatment groups showed significant changes in gene expression and pathways related to reproduction, immune function and carcinogenesis. Taken together, both BPA and BPS are potentially carcinogenic and have deleterious effects on the fertility of laying chickens by inducing inflammation, suggesting that BPS may not be a safe replacement for BPA.

Microbial Diversity and Community Variation in the Intestines of Layer Chickens.

The intestinal microbiota is increasingly recognized as an important component of host health, metabolism and immunity. Early gut colonizers are pivotal in the establishment of microbial community structures affecting the health and growth performance of chickens. White Lohmann layer is a common commercial breed. Therefore, this breed was selected to study the pattern of changes of microbiota with age. In this study, the duodenum, caecum and colorectum contents of white Lohmann layer chickens from same environment control farm were collected and analyzed using 16S rRNA sequencing to explore the spatial and temporal variations in intestinal microbiota. The results showed that the diversity of the microbial community structure in the duodenum, caecum and colorectum increased with age and tended to be stable when the layer chickens reached 50 days of age and the distinct succession patterns of the intestinal microbiota between the duodenum and large intestine (caecum and colorectum). On day 0, the diversity of microbes in the duodenum was higher than that in the caecum and colorectum, but the compositions of intestinal microbes were relatively similar, with facultative anaerobic Proteobacteria as the main microbes. However, the relative abundance of facultative anaerobic bacteria (Escherichia) gradually decreased and was replaced by anaerobic bacteria (Bacteroides and Ruminococcaceae). By day 50, the structure of intestinal microbes had gradually become stable, and Lactobacillus was the dominant bacteria in the duodenum (41.1%). The compositions of dominant microbes in the caecum and colorectum were more complex, but there were certain similarities. Bacteroides, Odoribacter and Clostridiales vadin BB60 group were dominant. The results of this study provide evidence that time and spatial factors are important factors affecting the intestinal microbiota composition. This study provides new knowledge of the intestinal microbiota colonization pattern of layer chickens in early life to improve the intestinal health of layer chickens.

Transcriptome Response of Liver and Muscle in Heat-Stressed Laying Hens.

Exposure to high ambient temperature has detrimental effects on poultry welfare and production. Although changes in gene expression due to heat exposure have been well described for broiler chickens, knowledge of the effects of heat on laying hens is still relatively limited. In this study, we profiled the transcriptome for pectoralis major muscle (n = 24) and liver (n = 24), during a 4-week cyclic heating experiment performed on layers in the early phase of egg production. Both heat-control and time-based contrasts were analyzed to determine differentially expressed genes (DEGs). Heat exposure induced different changes in gene expression for the two tissues, and we also observed changes in gene expression over time in the control animals suggesting that metabolic changes occurred during the transition from onset of lay to peak egg production. A total of 73 DEGs in liver were shared between the 3 h heat-control contrast, and the 4-week versus 3 h time contrast in the control group, suggesting a core set of genes that is responsible for maintenance of metabolic homeostasis regardless of the physiologic stressor (heat or commencing egg production). The identified DEGs improve our understanding of the layer's response to stressors and may serve as targets for genetic selection in the future to improve resilience.

Focal duodenal necrosis in chickens: attempts to reproduce the disease experimentally and diagnostic considerations.

Focal duodenal necrosis (FDN) is an intestinal disease of egg-layer chickens characterized by multifocal necrosis of the duodenal loop and proximal jejunum. Affected flocks usually have decreased egg weights and drops in egg production. Previous studies have associated this condition with Clostridium perfringens infection. We tried to reproduce FDN by experimental infection of egg-laying chickens using different netB-positive and netB-negative C. perfringens strains, and duodenal homogenate obtained from FDN lesions. Chickens challenged with C. perfringens and/or duodenal homogenate developed duodenitis after challenge. Gross lesions included mucosal erosions, hyperemia, mucosal hemorrhages, and watery intestinal content. Microscopic lesions included mild enterocyte degeneration and necrosis, and mild-to-moderate hemorrhage and lymphoplasmacytic and heterophilic infiltration of the lamina propria. Two netB-positive C. perfringens strains closely related to necrotic enteritis pathogenic strains, by genomic composition, were re-isolated from lesions. Necrosis of intestinal crypts was observed in chickens challenged with duodenal homogenate with or without C. perfringens coinfection. Characteristic microscopic FDN lesions with significant necrosis and loss of villus enterocytes were not reproduced.

Calidad durante el almacenamiento de huevos de gallinas alimentadas con dietas con aceite de palma / Egg quality during storage of eggs from hens fed diets with crude palm oil

RESUMEN Objetivo. Evaluar el efecto del aceite crudo de palma (ACP) sobre la calidad del huevo almacenado por varios días a diferentes temperaturas. Materiales y métodos. Un total de 240 gallinas de 28 semanas se alimentaron con dietas con 30g/kg de aceite de soya (AS) o ACP. Después de 12 semanas, muestras de huevos se almacenaron durante 0, 4, 8 y 12 días a 4, 12 y 24°C. Las características de calidad del huevo fueron evaluadas. Resultados. El huevo y la albúmina de gallinas en la dieta con ACP fueron más pesados que aquellos en la dieta con AS (p<0.05). La alta temperatura de almacenamiento redujo el peso del huevo, albúmina y yema, unidades Haugh (UH), altura de la albúmina y la yema, pH y color (p<0.05), pero aumentó el ancho de la albúmina y la yema, y la longitud de la albúmina (p<0.05). A medida que aumentaba el tiempo de almacenamiento, el peso del huevo, el peso y la altura de la albúmina, y la altura de la yema, el pH y el color se redujeron (p<0.05). Sin embargo, el peso y el ancho de la yema, el ancho y la longitud de la albúmina aumentaron (p<0.05). La interacción aceite x tiempo de almacenamiento (p<0.05) indico que la altura de la albumina, UH y el color de la yema de los huevos de la dieta con ACP fueron mejores a los 12 días de almacenamiento que en la dieta con AS. Conclusiones. Las gallinas en la dieta con ACP tuvieron huevos y albuminas más pesadas que las de las de la adieta con AS. La calidad del huevo disminuyó conforme el tiempo y temperatura de almacenamiento se incrementó, pero, los huevos de las gallinas suplementadas con ACP tuvieron mejor calidad en algunas características a los 12 días de almacenamiento.

ABSTRACT Objective. Evaluate the effect of crude palm oil (CPO) on quality traits of eggs stored various days at different temperatures. Material and Methods. A total of 240 hens, 28 weeks of age were fed diets with 30 g/kg of soybean oil (SO) or CPO. After 12 weeks, sample of eggs were stored during 0, 4, 8 and 12 days at 4, 12 and 24 °C. Egg quality traits were evaluated. Results. Egg and albumen from hens in CPO diet were heavier than those in SO (p<0.05). High storage temperature reduced egg, albumen and yolk weights, Haugh units (HU), albumen and yolk heights, pH and color (p<0.05), but increased albumen and yolk widths and albumen length (p<0.05). As storage time increased, egg weight, albumen weight and height, and yolk height, pH and colour were reduced (p<0.05). However, yolk weight and width, albumen width and length increased (p<0.05). Oil x storage time interaction (p<0.05) indicated that albumen height, HU and yellowness of yolk from hens in CPO diets were better at 12 days of storage than for hens fed SO. Conclusions. Hens in CPO diet had heavier eggs and albumen than those in SO diet. Egg quality traits decreased as temperature and days of storage increased, but, eggs from hens supplemented CPO had better quality in some traits at 12 days of storage.

A novel avian isolate of hepatitis E virus from Pakistan.

BACKGROUND: Avian hepatitis E virus (aHEV) has been associated with hepatitis-splenomegaly syndrome (HSS) in chickens along with asymptomatic subclinical infection in many cases. So far, four genotypes have been described, which cause infection in chickens, specifically in broiler breeders and layer chickens. In the present study, we isolated and identified two novel aHEV strains from the bile of layer chickens in Pakistan evincing clinical symptoms related to HSS. METHODOLOGY: Histology of liver and spleen tissues was carried out to observe histopathological changes in these tissues. Bile fluid and fecal suspensions were used for viral RNA isolation through MegNA pure and Trizol method which was further used for viral genome detection and characterization by cDNA synthesis and amplification of partial open reading frame (ORF) 1, ORF2 and complete ORF3. The bioinformatics tools; Molecular Evolutionary Genetics Analysis version 6.0 (MEGA 6), Mfold and ProtScale were used for phylogenic analysis, RNA secondary structure prediction and protein hydropathy analysis, respectively. RESULTS: Sequencing and phylogenetic analysis on the basis of partial methyltranferase (MeT), helicase (Hel) domain, ORF2 and complete ORF3 sequence suggests these Pakistani aHEV (Pak aHEV) isolates may belong to a Pakistani specific clade. The overall sequence similarity between the Pak aHEV sequences was 98-100%. The ORF1/ORF3 intergenic region contains a conserved cis-reactive element (CRE) and stem-loop structure (SLS). Analysis of the amino acid sequence of ORF3 indicated two hydrophobic domains (HD) and single conserved proline-rich domain (PRD) PREPSAPP (PXXPXXPP) with a single PSAP motif found in C-terminal. Amino acid changes S15 T, A31T, Q35H and G46D unique to the Pak aHEV sequences were found in the N-terminal region of ORF3. CONCLUSIONS: Our data suggests that Pak aHEV isolates may represent a novel Pakistani clade and high sequence homology to each other support the supposition they may belong to a monophyletic clade circulating in the region around Pakistan. The data presented in this study provide further information for aHEV genetic diversity, genotype mapping, global distribution and epidemiology.

Tetratrichomonas gallinarum granuloma disease in a flock of free range layers.

Granuloma disease in a flock of free range productive layers in the Netherlands in 2017 is described. The disease resembled granuloma outbreaks in layers caused by Tetratrichomonas gallinarum in 2013 and occurred in the same area in which the rearing farm considered as the source of the 2013 outbreaks was located. Between 55 and 84 weeks of age mortality was 20.3% (breeder's norm 3.9%). All dead hens examined (n = 20) showed granulomas especially in liver and ceca. Nine hens with or without liver and/or ceca granulomas were examined for trichomonads in mentioned organs by in situ hybridization (ISH), nested PCR, and cloning and sequencing. Ceca were also examined by culture. T. gallinarum ISH was positive in all livers and ceca with granulomas and negative in case granulomas were absent. T. gallinarum strain 13/16632, which caused the 2013 outbreaks was found in 4/8 hens with granulomas. Moreover, other trichomonads were detected: a T. gallinarum strain GPO-like and a Simplicimonas sp. strain GABC1-like. Mixed infections also occurred. Infectious causes of granuloma disease other than the afore-mentioned trichomonads could be excluded. Trichomonad DNA was not detected in environmental samples and wild ducks originating from the farm of concern, except for one duck in which the same Simplicimonas sp. as in hens was detected, leaving the source of the T. gallinarum infection in hens unknown. It is concluded that the herein described granuloma disease likely was caused by T. gallinarum strain 13/16632. However, the pathogenicity of the other trichomonads found remains to be clarified.

Pharmacokinetics of thymoquinone in layer chickens following oral and intravenous administration.

Thymoquinone (TQ) is the major constituent of Nigella sativa and known to possess a variety of pharmacological effects. This study was designed to evaluate the pharmacokinetic profile of TQ following oral (PO) and intravenous (IV) administration in layer chickens. The layer chickens were equally divided into two groups (six chickens in each group, total 12 chickens), and TQ was administered via PO and IV routes. For PO route, the dose was 20 mg/kg b.w. and for IV route, 5 mg/kg b.w. was administered, respectively. A sensitive and accurate High-Performance Liquid Chromatography (HPLC) technique was validated for the quantification of TQ from plasma. The limit of detection (LOD) and limit of quantification (LOQ) were 0.02 µg/ml and 0.05 µg/ml, respectively with >80% recovery. Maximum plasma concentration (Cmax ) following PO and IV administration was 8.805 and 4.497 µg/ml, respectively, while time to reach at maximum concentration (Tmax ) was 1 and 0.1 hr, respectively. The elimination half-lives were recorded as 1.02 and 0.978 hr, whereas the mean residence times were 1.79 and 1.036 hr following both PO and IV administration, respectively. The 85% PO bioavailability was indicative that TQ could be used for various therapeutic purposes in layer chickens.

Detection of gastrointestinal parasites in small-scale poultry layer farms in Leyte, Philippines.

BACKGROUND: Gastrointestinal (GIT) parasites can affect poultry productivity by compromising its health. It is well studied in other countries, but the documented reports in the Philippines have been limited. AIM: The aim of the present study was to evaluate the presence of GIT parasites in selected small-scale poultry layer farms in Leyte, Philippines. MATERIALS AND METHODS: A total of 243 stool samples from eight small-scale poultry layer farms in Leyte, Philippines, were examined for GIT parasites using floatation and sedimentation technique. Profile parameters were also obtained. Fecal samples were collected and analyzed using floatation and sedimentation techniques. Statistical significance between GIT parasite positivity and profile parameters was determined using Chi-square test. RESULTS: GIT parasites were detected in 92.2% of the samples (24.7% with single infection, 42.0% with 2-3 parasites, and 25.5% with three or more parasites). The common parasites detected were Ascaridia spp. (41.2%), Heterakis spp. (59.3%), Capillaria spp. (10.7%), Eimeria spp. (43.2%), and Strongyloides spp. (74.1%). Some profile parameters, including farm location, years in business, number of workers, nearby water system, the practice of fecal cleaning, and presence of other animals, were found to be significantly associated with GIT positivity. CONCLUSION: GIT parasites were detected in the poultry of small-scale layer farms in selected areas in Leyte, Philippines. This finding calls for the importance of routine GIT parasite monitoring and the implied need for regular deworming or dewormer rotation in the area.