Effects of peppermint essential oil and artifier on growth performance, carcass characteristics and nutrient digestibilities in broiler chickens fed with low energy diets.

The aim of this experiment was to investigate the effects of peppermint essential oil (PEO) and an emulsifier called artifier on growth performance, carcass characteristics, and nutrient digestibility of broiler chickens fed low-energy diets. A total of 240 one-day-old male Ross 308 broiler chickens were divided into five treatments: control, low-energy diet, low-energy diet + 150 ppm PEO, low-energy diet + 300 ppm artifier, and low-energy diet + 150 ppm PEO + 300 ppm artifier. Each treatment was replicated four times in a completely randomized design. The chickens were fed these treatments during the starter, grower, and finisher periods for a total of 42 days. The results indicated that broiler chickens receiving the low-energy diet supplemented with PEO + artifier had similar body weight gain, feed conversion ratio, and breast percentage, but exhibited significantly lower abdominal fat percentage compared to those fed the control diet (P < 0.05). Furthermore, birds receiving PEO and artifier in low-energy diets, either individually or in combination, demonstrated higher digestibility of dry matter and fat during the starter and grower periods in comparison to those receiving the low-energy diet without any additives (P < 0.05). Over the entire rearing period, the simultaneous inclusion of 150 ppm PEO and 300 ppm artifier in the low-energy diet resulted in comparable growth performance to the control diet. Additionally, the concurrent use of PEO and artifier in the low-energy diet for broiler chickens led to a decrease in abdominal fat, increased digestibility of fat and dry matter, and enhanced nutrient utilization compared to the control diet.

Genome-wide scans for selection signatures in indigenous chickens reveal candidate genes associated with local adaptation.

Population growth and climate change pose challenges to the sustainability of poultry farming. The emphasis on high-yield traits in commercialized breeds has led to a decline in their adaptability. Chicken varieties adapted to the local environment, possessing traits that facilitate adaptation to climate change, such as disease resistance and tolerance to extreme weather conditions, can improve hybridization outcomes. In this study, we conducted an analysis of the population structure and genetic diversity of 110 chickens representing indigenous breeds from southern China and two different commercial breeds. Further, we performed comparative population genomics, utilizing nucleotide diversity and fixation statistics, to characterize genomic features of natural selection and to identify unique genetic traits and potential selection markers developed by indigenous breeds after adapting to the local environment. Results based on genetic diversity and population structure analyses showed that indigenous varieties exhibited high levels of genetic diversity. Commercial breeds that have been indigenously bred demonstrated higher levels of genetic diversity than those that have not, and breeds with different selection practices displayed significant differences in genetic structure. Additionally, we further searched for potential genomic regions in native chicken ecotypes, uncovering several candidate genes related to ecological adaptations affecting local breeds, such as IKBKB, S1PR1, TSHR, IL1RAPL1 and AMY2A, which are involved in disease resistance, heat tolerance, immune regulation and behavioral traits. This work provides important insights into the genomic characterization of ecotypes of native chicken in southern China. The identification of candidate genes associated with traits such as disease resistance, heat tolerance, immunomodulation, and behavioral traits is a significant outcome. These candidate genes may contribute to the understanding of the molecular basis of the adaptation of the southern native chicken to the local environment. It is recommended that these genes be integrated into chicken breeding programs to enhance sustainable agriculture and promote effective conservation and utilization strategies.

Beyond probiotics, uses of their next-generation for poultry and humans: A review.

The production of healthy food is one of the basic requirements and challenges. Research efforts have been introduced in the human's food industry to reduce the microbial resistance and use safe and healthy alternatives with a high durability. However, the conducted work about these issues in the field of livestock animal production have been started since 2015. Inappropriate and extensive use of antibiotics has resulted in the increase of antimicrobial resistance, presence of drug residues in tissues, and destruction of the gut microbiome. Therefore, discovering and developing antibiotic substitutes were urgent demands. Probiotic compounds containing living micro-organisms are important antibiotic alternative that have been beneficially and extensively used in humans, animals, and poultry. However, some probiotics show some obstacles during production and applications. Accordingly, this review article proposes a comprehensive description of the next-generation of probiotics including postbiotics, proteobiotics, psychobiotics, immunobiotics and paraprobiotics and their effects on poultry production and human's therapy. These compounds proved great efficiency in terms of restoring gut health, improving performance and general health conditions, modulating the immune response and reducing the pathogenic micro-organisms. However, more future research work should be carried out regarding this issue.

An Interactive Feeder to Induce and Assess Emotions from Vocalisations of Chickens.

Understanding the emotional states of animals is a long-standing research endeavour that has clear applications in animal welfare. Vocalisations are emerging as a promising way to assess both positive and negative emotional states. However, the vocal expression of emotions in birds is a relatively unexplored research area. The goal of this study was to develop an interactive feeding system that would elicit positive and negative emotional states, and collect recordings of the vocal expression of these emotions without human interference. In this paper, the mechatronic design and development of the feeder is described. Design choices were motivated by the desire for the hens to voluntarily interact with the feeder and experience the different stimuli that were designed to induce (1) positive low-arousal, (2) positive high-arousal, (3) negative low-arousal, and (4) negative high-arousal states. The results showed that hens were motivated to engage with the feeder despite the risk of receiving negative stimuli and that this motivation was sustained for at least 1 week. The potential of using the interactive feeder to analyse chicken vocalisations related to emotional valence and arousal is being explored, offering a novel proof of concept in animal welfare research. Preliminary findings suggest that hens vocalised in response to all four stimulus types, with the number of vocalisations, but not the probability of vocalising, distinguishing between low- and high-arousal states. Thus, the proposed animal-computer interaction design has potential to be used as an enrichment device and for future experiments on vocal emotions in birds.

Effect of dietary methionine to crude protein ratio on performance of Ross 308 broiler chickens aged 22 to 42 days.

An experiment was conducted to determine the effect of dietary methionine to crude protein (CP) ratio on the performance of male Ross 308 broiler chickens aged 22 to 42 days. The diets were isocaloric and isonitrogenous but with different methionine-to-CP ratios. The diets, based on methionine to crude protein ratios, were M0.020 (0.020), M0.025 (0.025) M0.030 (0.030), M0.040 (0.040), or M0.045 (0.045). A complete randomized design was used. A quadratic type of equation was used to determine dietary methionine to CP ratios for optimal performance of the chickens. Dietary methionine to CP ratio had no effect (P > 0.05) on feed intake, live weight gain, live weight, feed efficiency, metabolizable energy intake, nitrogen retention, abdominal fat pad weight, breast meat nitrogen and methionine contents, and meat flavour and shear force values of the chickens, but it affected (P < 0.05) CP digestibility, carcass and breast weights, and breast meat tenderness and juiciness. Methionine to CP ratios of 0.039, 0.038, 0.050, and 0.050 were calculated to result in optimal CP digestibility, carcass weight, breast meat tenderness, and juiciness, respectively. These results may imply that dietary methionine to CP ratio requirements for broiler chickens will depend on the production parameter of interest.

Epigenetic Factor MicroRNAs Likely Mediate Vaccine Protection Efficacy against Lymphomas in Response to Tumor Virus Infection in Chickens through Target Gene Involved Signaling Pathways.

Epigenetic factors, including microRNAs (miRNAs), play an important role in affecting gene expression and, therefore, are involved in various biological processes including immunity protection against tumors. Marek's disease (MD) is a highly contagious disease of chickens caused by the MD virus (MDV). MD has been primarily controlled by vaccinations. MD vaccine efficacy might, in part, be dependent on modulations of a complex set of factors including host epigenetic factors. This study was designed to identify differentially expressed miRNAs in the primary lymphoid organ, bursae of Fabricius, in response to MD vaccination followed by MDV challenge in two genetically divergent inbred lines of White Leghorns. Small RNA sequencing and bioinformatic analyses of the small RNA sequence reads identified hundreds of miRNAs among all the treatment groups. A small portion of the identified miRNAs was differentially expressed within each of the four treatment groups, which were HVT or CVI988/Rispens vaccinated line 63-resistant birds and line 72-susceptible birds. A direct comparison between the resistant line 63 and susceptible line 72 groups vaccinated with HVT followed by MDV challenge identified five differentially expressed miRNAs. Gene Ontology analysis of the target genes of those five miRNAs revealed that those target genes, in addition to various GO terms, are involved in multiple signaling pathways including MAPK, TGF-ß, ErbB, and EGFR1 signaling pathways. The general functions of those pathways reportedly play important roles in oncogenesis, anti-cancer immunity, cancer cell migration, and metastatic progression. Therefore, it is highly likely that those miRNAs may, in part, influence vaccine protection through the pathways.

Relative biological efficacy of methionine hydroxy analogue-free acid compared to dl-methionine in the broiler chickens.

BACKGROUND: In the broiler's diets based on corn-soya bean meal, methionine (Met) and cystine (Cys), known as sulphur amino acids (SAAs), are the first limiting indispensable amino acids because of their limited presence, which are supplemented with different synthetic sources. Evaluation of the biological effectiveness of these sources can be important in their correct replacement, especially in the starter and growth diets. OBJECTIVES: The current study was done to assess the relative biological efficacy (RBE) of liquid Met hydroxy analogue-free acid (MHA-FA) in comparison with dl-Met (dl-Met) based on broiler performance traits at different levels of digestible SAA in the 1-11 (starter) and 11-25 (grower) days of age periods. METHODS: Two experiments were developed with treatments consisting of a basal diet without Met addition that met the nutrient and energy requirements of broilers with the exception of SAAs (Met + Cys) and five increasing Met doses for both sources (dl-Met and/or MHA-FA), resulting in digestible SAA concentrations from 0.62% to 1.02% of diet in the starter period (Trial 1) and 0.59% to 0.94% of diet in the grower period (Trial 2). The multi-linear regression model and slope ratio method were employed to calculate the RBE of MHA-FA compared with dl-Met for measured variables. RESULTS: In both experiments, the results obtained during the starter and grower periods with the different Met supplementations show significant growth responses to digestible SAAs levels. By increasing dietary dl-Met and/or MHA-FA levels, the growth performance traits and immune responses were improved (quadratic; p < 0.05). The RBE of MHA-FA compared to dl-Met on an equimolar basis was estimated 66%-89% (59%-79% on a weight-to-weight basis). CONCLUSIONS: It is concluded that the RBE of MHA-FA in comparison with dl-Met depends on broiler chicken age and what attribute is being evaluated.

Arginine alleviates Clostridium perfringens α toxin-induced intestinal injury in vivo and in vitro via the SLC38A9/mTORC1 pathway.

Introduction: Clostridium perfringens α toxin is a main virulence factor responsible for gut damage in animals. Arginine is a functional amino acid exhibiting significant immunoregulatory activities. However, the effects and immunoregulatory mechanisms of arginine supplementation on α toxin-induced intestinal injury remain unclear. Methods: In vivo, 256 male Arbor Acres chickens were randomly assigned to a 2×2 factorial arrangement, involving diet treatments (with or without 0.3% arginine supplementation) and immunological stress (with or without α toxin challenge). In vitro, IEC-6 cells were treated with or without arginine in the presence or absence of α toxin. Moreover, IEC-6 cells were transfected with siRNA targeting mTOR and SLC38A9 to explore the underlying mechanisms. Results and discussion: The results showed that in vivo, arginine supplementation significantly alleviated the α toxin-induced growth performance impairment, decreases in serum immunoglobulin (Ig)A and IgG levels, and intestinal morphology damage. Arginine supplementation also significantly reduced the α toxin-induced increase in jejunal proinflammatory cytokines interleukin (IL)-1ß, IL-6 and IL-17 mRNA expression. Clostridium perfringens α toxin significantly decreased jejunal mechanistic target of rapamycin (mTOR) and solute carrier family 38 member 9 (SLC38A9) mRNA expression, while arginine supplementation significantly increased mTOR and SLC38A9 mRNA expression. In vitro, arginine pretreatment mitigated the α toxin-induced decrease in cell viability and the increase in cytotoxicity and apoptosis. Arginine pretreatment also alleviated the α toxin-induced upregulation of mRNA expression of inflammation-related cytokines IL-6, C-X-C motif chemokine ligand (CXCL)10, CXCL11 and transforming growth factor-ß (TGF-ß), as well as apoptosis-related genes B-cell lymphoma-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-extra large (Bcl-XL) and cysteinyl aspartate specific proteinase 3 (Caspase-3) and the ratio of Bax to Bcl-2. Arginine pretreatment significantly increased the α toxin-induced decrease in mTOR, SLC38A9, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1 (4EBP1) and ribosomal protein S6 kinase (S6K) mRNA expression. Knockdown SLC38A9 and mTOR largely abrogated the positive effects of arginine pretreatment on α toxin-induced intracellular changes. Furthermore, SLC38A9 silencing abolished the increased mTOR mRNA expression caused by arginine pretreatment. In conclusion, arginine administration attenuated α toxin-induced intestinal injury in vivo and in vitro, which could be associated with the downregulation of inflammation via regulating SLC38A9/mTORC1 pathway.

Poultry Gastrointestinal-derived Lactic Acid Bacteria (pGIT-d-LAB) Inhibit Multiple Antibiotics Resistance Bacterial and Fungal Pathogens.

Background: To develop a probiotic formulation for poultry feed, a few poultry gastrointestinal derived lactic acid bacteria (pGIT-d-LAB) were isolated from chicken intestinal specimens and in vitro experiment was performed to evaluate their efficacy as potential probiotic candidate. Methods: A total of 6 strains of LAB: Lactobacillus brevis (L. brevis), Lactobacillus acidophilus (L. acidophilus), Lactobacillus casei (L. casei), Pediococci spp, Lactobacillus fermentum (L. fermentum) and Lactobacillus plantarum (L. plantarum) were isolated and cultured for collection of Cell Free Supernatant (CFS). CFS collected was tested against pathogenic bacterial isolated from chicken feces as well as prevalent fungal pathogens, utilizing agar-well diffusion techniques. A preliminary investigation into the susceptibility of the pathogens to diverse antibiotics and antifungal drugs was conducted. Bacterial pathogens exhibiting resistance to a minimum of three classes of antibiotics were subsequently identified for pGIT-d-LAB CFS screening. Results: The observed results revealed that the CFS derived from the isolates exhibited varying degrees of growth inhibition against different pathogens. Among the tested pGIT-d-LAB isolates, L. acidophilus demonstrated the most prominent zone of inhibition, measuring 18 mm against Klebsiella pneumoniae ZTAC 1233. Notably, Citrobacter diversus ZTAC 1255 showed resistance to all tested pGIT-d-LAB. Quantification of the metabolites produced was performed, and peak production levels was determined. L. acidophilus produced the highest amount of lactic acid (1.789g/l), Pediococci spp. produced the highest amount of diacetyl and H202 (1.918g/l) (0.0025g/l) at 48 hr peak values respectively. Conclusion: The test isolates are potential probiotic candidates for controlling pathogens in poultry.

Significant genomic introgression from grey junglefowl (Gallus sonneratii) to domestic chickens (Gallus gallus domesticus).

BACKGROUND: Chicken is one of the most numerous and widely distributed species around the world, and many studies support the multiple ancestral origins of domestic chickens. The research regarding the yellow skin phenotype in domestic chickens (regulated by BCO2) likely originating from the grey junglefowl serves as crucial evidence for demonstrating the multiple origins of chickens. However, beyond the BCO2 gene region, much remains unknown about the introgression from the grey junglefowl into domestic chickens. Therefore, in this study, based on whole-genome data of 149 samples including 4 species of wild junglefowls and 13 local domestic chicken breeds, we explored the introgression events from the grey junglefowl to domestic chickens. RESULTS: We successfully detected introgression regions besides BCO2, including two associated with growth trait (IGFBP2 and TKT), one associated with angiogenesis (TIMP3) and two members of the heat shock protein family (HSPB2 and CRYAB). Our findings suggest that the introgression from the grey junglefowl may impact the growth performance of chickens. Furthermore, we revealed introgression events from grey junglefowl at the BCO2 region in multiple domestic chicken breeds, indicating a phenomenon where the yellow skin phenotype likely underwent strong selection and was retained. Additionally, our haplotype analysis shed light on BCO2 introgression event from different sources of grey junglefowl into domestic chickens, possibly suggesting multiple genetic flows between the grey junglefowl and domestic chickens. CONCLUSIONS: In summary, our findings provide evidences of the grey junglefowl contributing to the genetic diversity of domestic chickens, laying the foundation for a deeper understanding of the genetic composition within domestic chickens, and offering new perspectives on the impact of introgression on domestic chickens.

Genetic insertion of mouse Myxovirus-resistance gene 1 increases innate resistance against both high and low pathogenic avian influenza virus by significantly decreasing replication in chicken DF1 cell line.

Avian influenza virus (AIV) is a constant threat to animal health with recent global outbreaks resulting in the death of hundreds of millions of birds with spillover into mammals. Myxovirus-resistance (Mx) proteins are key mediators of the antiviral response that block virus replication. Mouse (Mu) Mx (Mx1) is a strong antiviral protein that interacts with the viral nucleoprotein to inhibit polymerase function. The ability of avian Mx1 to inhibit AIV is unclear. In these studies, Mu Mx1 was stably introduced into chicken DF1 cells to enhance the immune response against AIV. Following infection, titers of AIV were significantly decreased in cells expressing Mu Mx1. In addition, considerably less cytopathic effect (CPE) and matrix protein staining was observed in gene-edited cells expressing Mu Mx1, suggesting Mu Mx1 is broadly effective against multiple AIV subtypes. This work provides foundational studies for use of gene-editing to enhance innate disease resistance against AIV.

Antimicrobial resistance patterns and genes of Campylobacter jejuni isolated from chickens in Pasuruan, Indonesia.

Background: Poultry is one of the most prominent sources of Campylobacter jejuni, which is also a major means of transmission to people. Campylobacter jejuni contamination in chicken meat comes from chicken feces because it naturally exists in the intestines of chickens. Aim: The purpose of this study is to identify the antibiotic resistance patterns and genes of C. jejuni, which was found in chickens in Pasuruan, Indonesia. Methods: The samples used in this study were 200 contents of the small intestine of broiler chickens from 40 farms in Pasuruan Regency. The enriched sample was streaked on the selective media of modified charcoal cefoperazone deoxycholate agar containing the CCDA selective supplement. Antimicrobial susceptibility test utilizing the Kirby-Bauer diffusion test method in accordance with Clinical and Laboratory Standards Institute standards. The polymerase chain reaction (PCR) method was used to detect the (hipO), which encodes the C. jejuni strain, fluoroquinolone resistance (gyrA), beta-lactam resistance (blaOXA-61), and tetracycline resistance (tetO) genes. Results: The findings revealed a 14% (28/200) prevalence of C. jejuni in the small intestine of broiler chickens. These isolates showed high resistance to enrofloxacin (92.9%). All isolates (100%) were susceptible to amoxicillin-clavulanate. The PCR results showed all C. jejuni isolates (100%) detected the gyrA gene, 96.4% detected the blaOXA-61 gene, and 50% detected the tetO gene. Conclusion: The findings of antimicrobial resistance at a high level from the small intestine of broiler chickens illustrate the potential threat to human health. To lessen the effects now and in the future, coordinated and suitable action is needed, as well as steps to guarantee the poultry industry's economic survival and public health insurance.

Effects of dietary water-soluble extract of rosemary supplementation on growth performance and intestinal health of broilers infected with Eimeria tenella.

This study was conducted to explore the effect of dietary supplementation of water-soluble extract of rosemary (WER) on growth performance and intestinal health of broilers infected with Eimeria tenella (E. tenella), and evaluate the anticoccidial activity of WER. 360 one-day-old Chinese indigenous male yellow-feathered broiler chickens were randomly allocated to six groups: blank control (BC) group and infected control (IC) group received a basal diet; positive control (PC) group, received a basal diet supplemented with 200 mg/kg diclazuril; WER100, WER200, and WER300 groups received a basal diet containing 100, 200, and 300 mg/kg WER, respectively. On d 21, all birds in the infected groups (IC, PC, WER100, WER200 and WER300) were orally gavaged with 1 mL phosphate-buffered saline (PBS) of 8×104 sporulated oocysts of E. tenella, and birds in the BC group were administrated an aliquto of PBS dilution. The results showed that dietary supplementation of 200 mg/kg WER increased average daily gain (ADG) of broilers compared to the IC group from d 22 to d 29 (P < 0.001). The anticoccidial index values of 100, 200 and 300 mg/kg WER were 137.49, 157.41 and 144.22, respectively, which indicated that WER exhibited a moderate anticoccidial activity. Compared to the IC group, the groups supplemented with WER (100, 200 and 300 mg/kg) significantly lowered fecal occyst output (P < 0.001) and cecal coccidia oocysts, alleviated intestinal damage and maintained the integrity of intestinal epithelium. Dietary supplementation with WER significantly improved antioxidant capacity, elevated the levels of secretory immunoglobulin A, and diminished inflammation within the cecum, particularly at a dosage of 200 mg/kg. The results of this study indicated that dietary supplementation with 200 mg/kg WER could improve broiler growth performance and alleviate intestinal damage caused by coccidiosis.

The Impact of Essential Amino Acids on the Gut Microbiota of Broiler Chickens.

The research involving the beneficial aspects of amino acids being added to poultry feed pertaining to performance, growth, feed intake, and feed conversion ratio is extensive. Yet currently the effects of amino acids on the gut microbiota aren't fully understood nor have there been many studies executed in poultry to explain the relationship between amino acids and the gut microbiota. The overall outcome of health has been linked to bird gut health due to the functionality of gastrointestinal tract (GIT) for digestion/absorption of nutrients as well as immune response. These essential functions of the GI are greatly driven by the resident microbiota which produce metabolites such as butyrate, propionate, and acetate, providing the microbiota a suitable and thrive driven environment. Feed, age, the use of feed additives and pathogenic infections are the main factors that have an effect on the microbial community within the GIT. Changes in these factors may have potential effects on the gut microbiota in the chicken intestine which in turn may have an influence on health essentially affecting growth, feed intake, and feed conversion ratio. This review will highlight limited research studies that investigated the possible role of amino acids in the gut microbiota composition of poultry.

Broiler Chicken Cecal Microbiome and Poultry Farming Productivity: A Meta-Analysis.

The cecal microbial community plays an important role in chicken growth and development via effective feed conversion and essential metabolite production. The aim of this study was to define the microbial community's variants in chickens' ceca and to explore the most significant association between the microbiome compositions and poultry farming productivity. The meta-analysis included original data from 8 control broiler chicken groups fed with a standard basic diet and 32 experimental groups supplemented with various feed additives. Standard Illumina 16S-RNA gene sequencing technology was used to characterize the chicken cecal microbiome. Zootechnical data sets integrated with the European Production Effectiveness Factor (EPEF) were collected. Analysis of the bacterial taxa abundance and co-occurrence in chicken cecal microbiomes revealed two alternative patterns: Bacteroidota-dominated with decreased alpha biodiversity; and Bacillota-enriched, which included the Actinomycetota, Cyanobacteriota and Thermodesulfobacteriota phyla members, with increased biodiversity indices. Bacillota-enriched microbiome groups showed elevated total feed intake (especially due to the starter feed intake) and final body weight, and high EPEF values, while Bacteroidota-dominated microbiomes were negatively associated with poultry farming productivity. The meta-analysis results lay the basis for the development of chicken growth-promoting feed supplementations, aimed at the stimulation of beneficial and inhibition of harmful bacterial patterns, where relevant metagenomic data can be a tool for their control and selection.

Evaluation of Clostridium autoethanogenum protein as a new protein source for broiler chickens in replacement of soybean meal.

Objective: The object of this study was to investigate the effect of replacing soybean meal with Clostridium autoethanogenum protein (CAP) in broiler diets on growth performance, blood indicators, antioxidant capacity, and immune function. Methods: A total of 180 Arbor Acres broilers were randomly divided into three treatments, each treatment with six replicates and 10 broilers per replicate for a 42-day feeding trial. The control group (CON) was fed corn-soybean meal based diet. The CAP-1 and CAP-2 groups were considered to use CAP to replace 25% or 50% of soybean meal in the diet, respectively. The average daily gain and average daily feed intake of broilers at 1-21 d, 22-42 d, and 1-42 d were measured, and the feed conversion ratio was calculated. At the 42nd day of age, two broilers with similar weights and fasted for 12h were selected in each replicate for blood collection from the brachial wing vein. The blood routine indicators, serum biochemical indicators, serum antioxidant capacity, and immunoglobulin content of broiler chickens were measured. Results: Replacement of soybean meal with 25% (CAP-1) and 50% (CAP-2) CAP significantly increased the average daily gain of 22-42 d and 1-42 d and decreased the average daily feed intake and feed conversion rate (p<0.05). The CAP-1 group, and CAP-2 group significantly increased hemoglobulin in the blood of broilers, while the CAP-2 group increased hematocrit content (p<0.05). Compared with the control group, the contents of superoxide dismutase and immunoglobulin A in serum of the CAP-2 group were significantly increased, while the contents of malondialdehyde in CAP group were significantly decreased (p<0.05). Conclusion: Replacing soybean meal with CAP led to significant improvements in the growth performance, antioxidant capacity, and immunoglobulin content of broilers.

Antimicrobial Resistance Profiles of Campylobacter spp. Recovered from Chicken Farms in Two Districts of Bangladesh.

The rapid emergence of antimicrobial resistance (AMR) in Campylobacter has reinforced its status as a foodborne pathogen of significant public health concern. Resistant Campylobacter is typically transferred to humans via the consumption of contaminated animal products, particularly poultry. The genes associated with antimicrobial resistance in Campylobacter spp. are poorly understood. To address this knowledge gap, we conducted a prevalence survey of AMR Campylobacter across 84 chicken farms in two districts of Bangladesh. Pooled cloacal swabs were collected from chickens and underwent bacteriological testing for Campylobacter spp. with PCR confirmation. Antimicrobial susceptibility was tested against 14 antibiotics by disk diffusion method, and 12 resistance genes were screened in Campylobacter-positive isolates using multiplex PCR. A total of 34 (40.5%) farms were Campylobacter-positive of which 73.5% of isolates were resistant to at least 10 antibiotics. The antimicrobial susceptibility results indicate a high level of resistance against streptomycin (97.1%), clindamycin (97.1%), ampicillin (94.1%), tetracycline (94.1%), erythromycin (91.2%), ciprofloxacin (88.2%), nalidixic acid (85.3%), and imipenem (82.4%), and comparatively a low frequency of resistance to chloramphenicol (47.1%), ceftazidime (44.1%), and colistin (35.3%). Multidrug-resistant (MDR) and extensively drug-resistant Campylobacter were identified in 97.1%, and 50% of isolates, respectively. Ten resistance genes were identified including blaTEM (in 97.1% of isolates), strA-strB (85.9%), tetA (70.6%), tetB (32.4%), qnrS (23.5%), blaCTX-M-1 (20.6%), qnrB (20.6%), blaSHV (8.8%), aadB (5.9%), and qnrA (2.9%). Our findings demonstrate that resistance to ampicillin, tetracycline, and ceftazidime in Campylobacter isolates was significantly (p ≤ 0.05) associated with the presence of blaTEM, tetA, and blaSHV genes, respectively. The high rates of AMR in Campylobacter isolates from our study are not surprising given the liberal use of antimicrobials and incomplete biosecurity provisions on farms. Of particular concern are resistance rates to those classes of antibiotics that should be reserved for human use (azithromycin, ciprofloxacin, and colistin). AMR was more prevalent in chicken farms that used multiple antibiotics, engaged in prophylactic treatment of the birds, and improperly disposed of antibiotic packages. The high prevalence of MDR in chicken-derived Campylobacter isolates from the different regions of our study reinforces the need for more prudent use of antimicrobial compounds in Bangladeshi chicken farms.

Effects of lighted incubation and foraging enrichment during rearing on individual fear behavior, corticosterone, and neuroplasticity in laying hen pullets.

Environmental conditions during incubation and rearing can influence stress responsivity of laying hen pullets throughout their lifespan, and therefore have important implications for their welfare. In this study, a 12:12h green LED light-dark cycle during incubation and larvae provisioning as enrichment during rearing were tested as strategies to optimize early-life conditions and thereby decrease stress responsivity in ISA Brown laying hens. A combination of parameters was measured to indicate neuronal, physiological, and behavioral changes that may affect fear and stress. The proteins calbindin D28k (calbindin1), doublecortin (DCX), and neuronal nuclein protein (NeuN) were quantified after hatch as a proxy for brain plasticity. Plasma and feather corticosterone levels were measured after hatch and at the end of the rearing phase, and fearfulness was investigated through a series of behavioral tests (i.e., voluntary approach, open field, tonic immobility, and manual restraint tests). No effects of light during incubation were found on calbindin1, DCX, or NeuN. Neither of the treatments affected corticosterone levels in blood plasma and feathers. Light-incubated pullets showed less fearfulness towards humans in the voluntary approach test, but not in the other behavioral tests reported in this study. Larvae provisioning had no effect on behavior. Our study showed minor effects of light during incubation and no effects of enrichment during rearing on stress responsivity of laying hen pullets. The small effects may be explained by the enriched rearing conditions for all birds in this experiment (low stocking density, natural daylight, and 24/7 classical music). Given the promising results of lighted incubation in other studies, which were mostly performed in broiler chickens, and evidence regarding the positive effects of enrichment during rearing, the potential of these strategies to improve laying hen welfare needs to be explored further.

Antibiotic resistance of Gallibacterium anatis biovar haemolytica isolates from chickens.

Introduction: Gallibacterium anatis is an opportunistic bacteria inducing a range of clinical signs in poultry. Gallibacterium anatis strains show multidrug resistance to antibacterial substances. The purpose of this study was to examine the susceptibility of G. anatis biovar haemolytica isolates collected from the respiratory, reproduction and gastrointestinal tracts of chickens to different antibiotics from various classes. Material and Methods: Gallibacterium anatis biovar haemolytica was identified in tracheal swab and gastrointestinal and reproductive tract tissue samples from Polish layer and broiler chicken flocks. Twenty six isolates with ß-haemolysis capability, each from a different flock, obtained from the respiratory (n = 8), reproductive (n = 10) and gastrointestinal (n = 8) tracts were selected and identified by matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry after culturing. A PCR method targeting the 16S genes was used for verification of isolates. The isolates' susceptibility to 20 antimicrobials was evaluated using the disc diffusion method for 8 drugs and the dilution method for the other 12. In addition, they were tested for the presence of the GtxA, gyrB and flfA virulence genes and blaROB, aphA, tetB and tetH antibiotic resistance genes by PCR. Results: The most prevalent antibiotic resistance was to tilmicosin, tylosin and quinupristin/dalfopristin (all 100%), erythromycin (96.2%), tetracycline (96.2%), linezolid (92.3%) and teicoplanin (92.3%). Universal susceptibility was to only one antibiotic, chloramphenicol. Statistically significant differences were found between the resistance of gastrointestinal tract strains and that of strains from other tracts to daptomycin, gentamicin, ciprofloxacin and colistin. The GtxA and gyrB genes were detected in 100% of isolates and flfA in 19.2%. The isolates most frequently contained tetB and less frequently tetH and aphA, and did not contain blaROB. Conclusion: Most G. anatis biovar haemolytica isolates were resistant to many classes of antibiotics. Therefore, it is necessary and important to be vigilant for the occurrence of these bacteria and thorough in their diagnosis.