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We examined the weight distribution of skeletal muscles of the red jungle fowl, then compared these values with those of domesticated populations to determine how muscle distribution has changed by selecting breeding. Sonia, Fayoumi, and Rhode Island Red were selected for comparison from livestock breeds, while Japanese Shamo and Thai fighting cocks were selected from cockfighting groups. Principal component analysis was applied using body size-free data. The mass distribution of muscles clearly differed between the wild, livestock, and cockfighting groups, demonstrating that muscle distribution has changed after selecting breeding, coupled with functional demands of each group. The red jungle fowl, which has the ability to fly, could be clearly distinguished from the flightless domesticated populations due to differences in flight pectoral muscle size. The cervical muscles in the wild population were smaller than in the domesticated groups; these do not contribute to flight. The gluteal muscles were larger in the fighting cock group, functionally coupled to their traditionally preferred upright posture. Wild bird populations typically exhibit reduced weight of their hind limbs, associated with flight, but as the red jungle fowl displays largely terrestrial behavior, these muscles are similar in arrangement and relative size to those of the livestock groups. We showed that the mass distribution pattern of skeletal muscles expresses selecting breeding strategy and clearly reflects the specific traits for each group.

Introduction: In Indian GDP, veterinary sector plays a major role in contribution wherein poultry meat plays good protein source. But the health status is less studied in case of country fowl. Aims and Objectives: Indian country fowl are highly infected with producing emaciation, reduced growth rate, highly morbidity with enteritis. After confirmation by gross and microscopy, samples undergone histopathology study using Haematoxylin and Eosin stain (H&E). Thereafter, the samples also processed for detection of carbohydrate by McManus method using PAS stain and mucoprotein was detected using congo red method. Results: Adult possesses carbohydrate, protein and fat. Histologically the mature, immature and gravid segments were differentiated whereas the histochemical techniques qualifies the biomacromolecules. The lipid droplet showed a clear vacuole preferably in and around the periphery portion. The glycogen distributed in an even manner. Subsequently, the protein was deposited mostly at the periphery in the body surfaces as well as in the egg. Keywords: Gallus domesticus, Cestode (Raillietina sp.), Bio-macromolecules, Histochemistry, Histopathology.

Avian adenoviruses are an extremely diversified group of pathogens that recently triggering a variety of problems for poultry production. In particular, Inclusion Body Hepatitis-Hydropericardium Syndrome (IBH-HPS), which has been observed in broiler birds from 3 to 6 weeks of age and is associated with anemia, hemorrhagic disorders, hydropericardium, and high mortality. The disease has been reported worldwide, and recently it was reported in many Egyptian farms, causing severe economic losses. Therefore, the current study aimed to isolate, and genetically type the most common Adenovirus serotypes associated with this syndrome in Egyptian farms. A total of 50 broiler chicken farms (3-6 weeks old) located in different Egyptian governorates were examined. Macroscopically, the diseased flock revealed hydropericardium, enlarged friable livers with ecchymotic hemorrhages, and varying mortality rates (1 to 7.5%). Histopathologically, severe diffuse necrotizing enteritis, hepatitis, pericarditis, and diffuse lymphoid depletion of the spleen were the most prominent lesions. Liver tissues and cloacal swabs were collected from all examined flocks for FAdVs detection by conventional polymerase chain reaction (PCR) targeting the L1 loop in the hexon gene. The PCR products were sequenced for typing of the detected viruses. It was found that 10 out of 50 flocks examined were PCR positive for FAdVs (20%). Phylogenetic analysis of the sequenced genes revealed that the obtained viruses clustered with reference strains belonging to FAdV type D and E serotype 2, 11, and 8a respectively. The isolation of both FAdV type D and FAdV type E were carried out on a primary cell culture chicken embryo liver cell (CEL) and the presence of these viruses was confirmed by PCR after the appearance of cytopathic effect (CPE). From this study, it could be concluded that both FAdVs types D and E are the most common adenoviruses circulating in poultry farms suffering from hydropericardium and inclusion body hepatitis.

The effectiveness of formaldehyde egg disinfection is well documented in literature despite its reported toxicity. This study focused on the need for an optimum formaldehyde concentration (FC) that significantly reduces microbial load with minimal damaging effect on egg viability and hatchability. Using a true experimental design, bacterial load on formaldehyde-treated (FT) and control groups of eggs and hatchability were compared. Gram-staining and biochemical tests identified five bacterial species: Escherichia coli, Enterobacter sp., Bacillus cereus, Staphylococcus aureus and Micrococcus sp. with the two coliform bacteria, E. coli and Enterobacter sp., dominating. Comparison of median differences of bacterial load on eggs before and after formaldehyde treatment by Wilcoxon Signed-Rank Test showed marginal significance in bacterial load reduction (Z = -2.016, P = 0.044). This difference was observed for bacterial load between the control group (CG) and the FT group with FC 30/20 ml/g (U = 3.0, P = 0.047). The hatchability of the CG differed significantly from four FT groups of eggs with FC 30/20 ml/g showing the highest level of significance [χ² (1) = 14.71; P = 0.0001]. A FC of 30/20 ml/g produced the best domestic fowl egg disinfection compared to other FCs and hatchability decreased with increasing formalin volume.

The study examined effects seasons have on diseases frequency and mortality of poultry birds in Owerri urban south-eastern Nigeria. The study therefore employed the use of secondary and primary data. Climatological data on rainfall, temperature, relative humidity and mortality of birds was collected from Nimet, Sam Mbakwe International cargo Airport, Imo State and ministry of livestock development Owerri respectively for a period of 16 years (2004-2019). Also 83 copies of questionnaire were used in the analysis. Results showed positive correlation between rainfall and relative humidity (0.517 and 0.747) and an R² of 39.98% with implies that, the higher the rainfall the higher the relative humidity, and by extension the higher the frequency of deadly poultry diseases (Coccidiosis, Gumboro, Fowl cholera, Fowl typhoid) and mortality especially for broilers as the cell immune response of layer birds in defending itself against diseases and viruses are greater than that of broilers. While Newcastle diseases, Fowl pox, Fowl typhoid, chronic respiratory diseases mostly affect birds in the dry season (November- March). While Newcastle diseases, Fowl pox, Fowl typhoid, chronic respiratory diseases mostly affect birds in the dry season (November- March).In conclusion, seasons of the year affect poultry and its production in particular in Owerri urban. It is recommended that farmers should make sure they adopt proper roofing methods, as well as maintain good sanitation/hygiene practices to minimize heat and the occurrence and spread of diseases which may adversely affect birds across seasons.

The avian alimentary tract has evolved into different histologic structures to accommodate the physical and chemical features of several food types and flight requirements. We compared the esophagus, proventriculus, and gizzard of the domestic fowl,Gallus gallus domesticus(GGD) and kestrels,Falco tinnunculus(FT) using immunohistochemistry and scanning electron microscopy with various stains and lectins [Dolichos biflorusagglutinin (DBA) andRicinus communisagglutinin I (RCA120)], andα-smooth muscle actin (α-SMA). The esophagus of GGD demonstrated thickened epithelium, muscularis mucosae, and inner circular longitudinal tunica muscularis layers; moderate outer longitudinal tunica muscularis layers; and a true crop. In contrast, the esophagus of FT showed a thin epithelium, no muscularis mucosae, moderate inner longitudinal and thick outer circular tunica muscularis layers, and no true crop. In the proventriculus, the nature of the secretion in GGD was neutral, but that of FT was acidic and neutral. In the gizzard, the muscle coat of GGD byα-SMA had no muscularis mucosae, unlike FT, which had muscularis mucosae. In summary, there are many histologic differences between GGD and FT to meet their different physiologic needs, such as feeding.

The purpose of the research is analyzing ways and factors of trichinellosis causative agent Trichinella pseudospiralis transmitted in the Russian Federation. Materials and methods. The main trichinellosis monitoring stages, methods of veterinary and sanitary examination for trichinellosis, and parameters for neutralization of the pathogen are given. Results and discussion. The most likely circulation patterns of trichinellosis pathogen T. pseudospiralis in natural and synanthropic biocenoses, and the key links that ensure the activity of infection foci are presented.

Aim: The study was designed to elucidate the effects of quercetin in an extender on oxidative stress, mitochondrial activity and quality of Indian red jungle fowl (Gallus gallus murghi) sperm durin...

In fact, Free radical oxidation is one of the required factors of homeostasis as far as the active forms of oxygen, which have unpaired electron, perform useful functions, they are necessary intermediaries of numerous processes of normal cells’ functioning such as biosynthesis of prostaglandins and leukotriene as well as other biologically active substances. Also, they take part in pathological processes such as: inflammation, fever and others. Free radical oxidation is the process of direct oxygen transferring to the substrate forming peroxide, ketones, aldehydes, etc. Some amino acids, proteins, water and carbohydrates prone to the processes of peroxidation, but crucial place is granted to lipids at the expanse of fatty acids which are included to their composition. The article highlights the information concerning the analysis of the state of antioxidant system of hens in the process of feeding them with subtoxic doses of fungicide of Derosal (BAYER, Germany) (0.1 LD50 for a fowl, 900 mg/kg of the body mass) which active ingredient is benzimidazole carbamate (carbendazim, BMC). It presents the changes of these indicators in the process of using pharmacological and natural antioxidants. The analysis has figured out that giving Derosal to the hens without corrections (II group) has been followed with the activation of the process of lipids’ peroxidation (POL) in the blood plasma, that is followed with the increase of concentration of primary and secondary lipoperoxidation products – diene conjugates (DC) and TBA-active products on the average up to 21.0–25.0 %, catalase activity – up to 38.5 % and decrease of superoxide dismutase activity (SOD) up to 21.8 % and АОА – up to 17.8 %, that shows the adverse effects of pesticides on the system of antioxidant protection of the organism. In the fowls’ groups III, IV and V the adding of antioxidant E-selenium, ascorbic acid and thermally untreated buckwheat seeds as the source of quercetins and anthocyanins has led to the induction of antioxidant resources that is shown by the decrease in comparison with the control of content of diene conjugates, malonic dialdehyde and catalase (in average up to 13.6–33.6–18.0 % accordingly) and compensatory strengthening of superoxide dismutase activity and general activity of antioxidant (АОА) on the average up to 44.9–72.1–76.5 % and 15.3–13.4–21.7 % accordingly. In comparison with group II the activity of catalase has been lower as well as the activity of superoxide dismutase has been higher in all the groups almost in 1.5–2 times (Р ≤ 0.001).

The prototype fowl glioma-inducing virus (FGVp) causes fowl glioma and cerebellar hypoplasia in chickens. In this study, we investigated whether a strain of avian leukosis virus (ALV), associated w...

Many studies have been carried out to investigate the morphological structure of the syrinx in many bird species. However, the cellular organization of the syrinx in the fowls and pigeons is still unclear. The current study revealed that in fowl and pigeon, the syrinx is formed of three main parts including tympanum (cranial) part, intermediate syringeal part, and bronchosyringeal (caudal) part, in addition to pessulus and tympaniform membranes. A great variation in the structural characteristics of syrinx of fowl and pigeon was recorded. In fowl, the tympaniform membranes showed a characteristic distribution of elastic and collagen fibers which increase the elasticity of tympaniform membranes. Moreover, the bony pessulus helps the medial tympaniform membranes to be stiffer, vibrate more strongly so that louder sound will be generated. In pigeon, the lateral tympaniform membrane is of greater thickness so that the oscillation of this membrane is reduced and the amplitude is lower. Moreover, the pessulus is smaller in size and is formed mainly of connective tissue core (devoid of cartilaginous or bony plates), resulting in the failure of stretching and vibrating of the medial tympaniform membranes, that leads to the generation of deeper sound. Electron microscopic examination of the syringes of fowls and pigeons revealed numerous immune cells including dendritic cells, plasma cells, mast cells, and lymphocytes distributed within syringeal mucosa and invading the syringeal epithelium. Telocytes were first recorded in the syrinx of fowls and pigeons in this study. They presented two long telopodes that made up frequent close contacts with other neighboring telocytes, immune cells, and blood capillaries.

The domestic fowl, as a species, is considered a unique experimental model in immunological research where the humoral and cellular components of the immune response can be delineated by selective surgical bursectomy of newly hatched chicks combined with sublethal whole-body X-irradiation. Search for an effective, safe, and a short-acting general anesthetic for application in such young birds is, therefore, essential. Ketamine is a phencyclidine derivative with analgesic and anesthetic properties in both mammalian and avian species. Studies on its efficacy on young or naturally small birds, however, are apparently lacking. Our previous studies indicated that dose levels as high as 51 mg/kg and 37.2 mg/kg are not lethal to 2-day- and 6-day-old Hisex White domestic fowl chicks when given intramuscularly, respectively. Results of the present study show that the onset of ketamine anesthesia varied between 40 - 98 seconds in 2-day-old chicks of this breed depending upon dose level. The dose and response are inversely related (y = -19.3x + 107.5) where “y” is the time for onset of anesthesia in seconds and “x” is the dose as mg/bird. The time for recovery from anesthesia also varied (6.6 - 74.7 min) depending upon dose level. The dose and response, however, are positively correlated (y = 20.92x - 21.15). Doses ranging between 43.77 to 47.74 mg/kg are considered optimum to produce anesthesia with onset in 74 - 77 sec and recovery within 12 - 15 min. It is concluded that Ketamine has the potential for application as a general anesthetic for domestic fowl chicks for bursectomy. Critical clinical tests are necessary to evaluate further the performance of this drug when administered to young chicks.

Every year, billions of egg layer chicks around the world are hatched under highly stressful, industrial circumstances. Here, it is investigated how the stressful procedure in the commercial hatchery, including incubation, hatching, processing, and transport affects the chicks with regards to traits relevant for the egg production industry. These traits were compared to those of a control group hatched in a small incubator and handled gently och quietly in a quiet room without any processing and transport. The chicks were weighed at hatch and at eight additional time points: 4 days, 1 week (w), 2 w, 3 w, 5 w, 8 w, 20 w and 25 w of age. Feather pecking was studied at 15 w of age and damages to the feathers and injuries on the comb and wattle were assessed at 25 w of age. From 19 w of age, eggs were collected on three days per week, counted and weighed. Chicks from a commercial hatchery had a lower hatch weight than control chicks (pFunding: FORMAS [2016-01728]

Single nucleotide polymorphisms (SNPs) are widely used in genome-wide association studies and population genetics analyses. Next-generation sequencing (NGS) has become convenient, and many SNP-calling pipelines have been developed for human NGS data. We took advantage of a gap knowledge in selecting the appropriated SNP calling pipeline to handle with high-throughput NGS data. To fill this gap, we studied and compared seven SNP calling pipelines, which include 16GT, genome analysis toolkit (GATK), Bcftools-single (Bcftools single sample mode), Bcftools-multiple (Bcftools multiple sample mode), VarScan2-single (VarScan2 single sample mode), VarScan2-multiple (VarScan2 multiple sample mode) and Freebayes pipelines, using 96 NGS data with the different depth gradients of approximately 5X, 10X, 20X, 30X, 40X, and 50X coverage from 16 Rhode Island Red chickens. The sixteen chickens were also genotyped with a 50K SNP array, and the sensitivity and specificity of each pipeline were assessed by comparison to the results of SNP arrays. For each pipeline, except Freebayes, the number of detected SNPs increased as the input read depth increased. In comparison with other pipelines, 16GT, followed by Bcftools-multiple, obtained the most SNPs when the input coverage exceeded 10X, and Bcftools-multiple obtained the most when the input was 5X and 10X. The sensitivity and specificity of each pipeline increased with increasing input. Bcftools-multiple had the highest sensitivity numerically when the input ranged from 5X to 30X, and 16GT showed the highest sensitivity when the input was 40X and 50X. Bcftools-multiple also had the highest specificity, followed by GATK, at almost all input levels. For most calling pipelines, there were no obvious changes in SNP numbers, sensitivities or specificities beyond 20X. In conclusion, (1) if only SNPs were detected, the sequencing depth did not need to exceed 20X; (2) the Bcftools-multiple may be the best choice for detecting SNPs from chicken NGS data, but for a single sample or sequencing depth greater than 20X, 16GT was recommended. Our findings provide a reference for researchers to select suitable pipelines to obtain SNPs from the NGS data of chickens or nonhuman animals.

The rapid emergence of virulent and multidrug-resistant (MDR) non-typhoidal Salmonella (NTS) enterica serovars are a growing public health concern globally. The present study focused on the assessment of the pathogenicity and antimicrobial resistance (AMR) profiling of NTS enterica serovars isolated from chicken processing environments at wet markets in Dhaka, Bangladesh. A total number of 870 samples consisting of carcass dressing water (CDW), chopping board swabs (CBS), and knife swabs (KS) were collected from 29 wet markets. The prevalence of Salmonella was found to be 20% in CDW, 19.31% in CBS and 17.58% in KS, respectively. Meanwhile, the MDR Salmonella was found to be 72.41%, 73.21% and 68.62% in CDW, CBS, and KS, respectively. All isolates were screened by polymerase chain reaction (PCR) for eight virulence genes, namely invA, agfA, IpfA, hilA, sivH, sefA, sopE, and spvC. The S. Enteritidis and untyped Salmonella isolate harbored all virulence genes while S. Typhimurium isolates carried six virulence genes except sefA and spvC. Phenotypic resistance revealed decreased susceptibility to ciprofloxacin, streptomycin, ampicillin, tetracycline, gentamycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid and azithromycin. Genotypic resistance showed higher prevalence of plasmid mediated blaTEM followed by tetA, sul1, sul2, sul3, and strA/B genes. Harmonic and symmetrical trend was observed among the phenotypic and genotypic resistance patterns of the isolates. The research findings anticipate that MDR and virulent NTS enterica serovars are prevailing in the wet market environments which can easily enter into the human food chain. There was a resilient and significant correlation existent among the phenotypic and genotypic resistance patterns and virulence genes of Salmonella isolate recovered from carcass dressing water, chopping board swabs, and knife swabs (p < 0.05), respectively.

Chagas disease is mainly transmitted by triatomine insect vectors that feed on vertebrate blood. The disease has complex domiciliary infestation patterns and parasite transmission dynamics, influenced by biological, ecological, and socioeconomic factors. In this context, feeding patterns have been used to understand vector movement and transmission risk. Recently, a new technique using Liquid chromatography tandem mass spectrometry (LC-MS/MS) targeting hemoglobin peptides has showed excellent results for understanding triatomines’ feeding patterns. The aim of this study was to further develop the automated computational analysis pipeline for peptide sequence taxonomic identification, enhancing the ability to analyze large datasets data. We then used the enhanced pipeline to evaluate the feeding patterns of Triatoma dimidiata, along with domiciliary infestation risk variables, such as unkempt piles of firewood or construction material, cracks in bajareque and adobe walls and intradomiciliary animals. Our new python scripts were able to detect blood meal sources in 100% of the bugs analyzed and identified nine different species of blood meal sources. Human, chicken, and dog were the main blood sources found in 78.7%, 50.4% and 44.8% of the bugs, respectively. In addition, 14% of the bugs feeding on chicken and 15% of those feeding on dogs were captured in houses with no evidence of those animals being present. This suggests a high mobility among ecotopes and houses. Two of the three main blood sources, dog and chicken, were significantly (p < 0.05) affected by domiciliary infestation risk variables, including cracks in walls, construction material and birds sleeping in the intradomicile. This suggests that these variables are important for maintaining reproducing Triatoma dimidiata populations and that it is critical to mitigate these variables in all the houses of a village for effective control of these mobile vectors.

Fowl adenoviruses (FAdVs) are a large group of viruses of different serotypes. They are responsible for inclusion body hepatitis, adenoviral gizzard erosion, and hepatitis hydropericardium syndrome. The present study presents a comprehensive overview of FAdVs in Egypt, with a focus on the epidemiological features of virus serotypes across the country. We conducted molecular investigation of multiple FAdV species based on the genetic signature of hypervariable regions 1–4 in the loop1 (L1) region of the hexon gene. Epidemiologically, the Nile Delta governorates showed high positivity of FAdVs, which were more commonly found in broilers than in layers. Genetically, species D and serotype 8a/E dominated, and the findings also revealed the emergence of new FAdV serotypes 1, 3, and 8b. The comparative analysis of hypervariable regions in the L1 region of the hexon gene revealed variables specific to each virus serotype. In silico predictions of L1 region revealed variations in the molecular structure and predicted the antigenic epitopes which may affect the cross-antigenicity between the different FAdV species and serotypes., Fowl adenovirus, Loop1, HVRs, Egypt.

Magnolol is a bioactive polyphenolic compound commonly found in Magnolia officinalis. The aim of this study is to clarify the contribution of the magnolol additive on the growth performance of Linwu ducklings aging from 7 to 28 d, comparing to the effects of antibiotic additive (colistin sulphate). A total of 325, 7-d-old ducklings were assigned to 5 groups. Each group had 5 cages with 13 ducklings in each cage. The ducklings in different groups were fed with diets supplemented with 0, 100, 200 and 300 mg/kg magnolol additive (MA) (Control, MA100, MA200 and MA300) and 30 mg/kg colistin sulphate (CS30) for 3 weeks, respectively. Parameters regarding to the growth performance, intestinal mucosal morphology, serum biochemical indices, antioxidant and peroxide biomarkers and the expression levels of antioxidant-related genes were evaluated by one way ANOVA analysis. The results showed that 30 mg/kg colistin sulphate, 200 and 300 mg/kg magnolol additive improved the average final weight (P = 0.045), average daily body weight gain (P = 0.038) and feed/gain ratios (P = 0.001) compared to the control group. 200 and 300 mg/kg magnolol additive significantly increased the villus height/crypt depth ratio of ileum, compared to the control and CS30 groups (P = 0.001). Increased serum level of glucose (P = 0.011) and total protein (P = 0.006) were found in MA200 or MA300 group. In addition, comparing to the control and CS30 groups, MA200 or MA300 significantly increased the levels of superoxide dismutase (P = 0.038), glutathione peroxidase (P = 0.048) and reduced glutathione (P = 0.039) in serum. Moreover, the serum and hepatic levels of 8-hydroxy-2’-deoxyguanosine (P = 0.043 and 0.007, respectively) were lower in all MA groups compared to those of the control and CS30 group. The hepatic mRNA expression levels of superoxide dismutase-1, catalase and nuclear factor erythroid-2-related factor 2/erythroid-derived CNC-homology factor were also increased significantly in MA200 and MA300 groups (P < 0.05). Taken together, these data demonstrated that MA was an effective feed additive enhancing the growth performance of Linwu ducklings at 7 to 28 d by improving the antioxidant and intestinal mucosal status. It suggested that MA could be a potential ingredient to replace the colistin sulphate in diets.

Fowl adenoviruses (FAdVs), which are distributed worldwide, have caused considerable economic losses to poultry farms. Co-infection with FAdVs and other avian pathogens has been reported previously. However, the pathogenicity of different serotypes of FAdVs causing co-infection remains unclear. Herein, strain HN from FAdV species C serotype 4 (FAdV-4) and strain AH720 from species E serotype 8a (FAdV-8a) were used to assess the pathogenicity of their co-infection in specific-pathogen-free (SPF) chickens. Compared with chickens infected with FAdV-4 alone, those co-infected with FAdV-4 and FAdV-8a showed similar clinical symptoms, mortality rates and degree of tissue lesions, and notably decreased viral loads of HN. Conversely, the viral loads of AH720 increased markedly in the co-infection group compared with that in chickens infected with AH720 strain alone. Increased viral loads of AH720 in the liver were suspected to contribute to the pathogenicity of chickens co-infected with the HN and AH720 strains. This was further investigated by histopathology and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining analyses. Collectively, these data indicated that co-infection with FAdV-4 and FAdV-8a suppresses the replication and proliferation of FAdV-4 but enhances the replication and proliferation of FAdV-8a in chicken liver. This study will provide valuable information for the further investigation of the interactions between FAdV-4 and FAdV-8a during co-infection.

Red grape (Vitis vinifera L.) pomace’s (RGP) beneficial bioactive compounds could improve growth and meat quality traits in chickens and thus valorize RGP waste that is usually disposed in landfills to the detriment of the environment. This study investigated the effect of RGP inclusion in diets of Hy-line Silver Brown cockerels on physiological and meat quality responses. Five isonitrogenous and isocaloric diets were formulated by mixing a standard grower diet with RGP at 0 (G0), 15 (G15), 30 (G30), 45 (G45) and 60 g/kg (G60). A total of 250, 5-week-old cockerels (304.6 ± 6.57 g live-weight) were evenly allocated to 25 pens replicated 5 times per experimental diet. No linear and quadratic trends (P > 0.05) were observed for overall feed intake, body weight gain, feed conversion ratio, and meat quality traits as dietary RGP levels increased. Erythrocytes linearly decreased (P < 0.05), whereas mean corpuscular hemoglobin and urea linearly increased (P < 0.05) with RGP levels. There were significant quadratic effects for glucose, phosphorus, total protein, albumin, globulin, and cholesterol, from which a maximum RGP inclusion level was calculated to be 43 g/kg. In conclusion, dietary red grape pomace had no adverse effect on physiological parameters and meat quality traits of Hy-line Silver Brown cockerels. However, including red grape pomace beyond 43 g/kg could compromise serum biochemical parameters of the birds.

In this study fowl poxvirus has been isolated from naturally infected birds. This virus induced respiratory distress, 2, 3 and 4 days post inoculation, in intratracheally inoculated 8 week old chickens, these respiratory distress include congestion of tracheal mucosa, increased tracheal secreation and whitish-yellowish pastules appeared at ninth day post inoculation in the upper part of the trachea forming diphtheritic or pseudodiphtheritic membrane. Using scanning electron microscopy to study the changes that occurs in the epithelial surface of trachea. It was found that the virus causes proliferation of epithelial cells forming cluster like papilloma, deciliation and activation of goblet cells.

Outbreaks of inclusion body hepatitis have emerged in Morocco since 2013 and has resulted in significant economic losses to poultry farms. Three isolates of the causative virus, Fowl adenonovirus (FAdV)were characterized from chickens with IBH, but their pathogenicity has never been investigated. In this work, the pathogenicity of an isolate FAdV 11 (MOR300315 strain) was evaluated by inoculating a group of 40 SPF chickens at 3 days of age by oral route. A group of 40 chicks injected with phosphate-buffered saline solution was used as a control group. The infected chickens showed decreased weight gain from 3dpi. Necropsy displayed pallor and enlargement in liver, swelling and slight hemorrhage in kidney and spleen at 6 dpi. Histopathological changes were mainly characterized by severe and extensive hepatic necrosis associated with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The FAdV was reisolated in chicken embryo fibroblast cell culture from liver tissue homogenate of infected chicken from 3 to 6 dpi. Viral DNA was detected by PCR in liver, kidney, spleen and cloacal swabs from 3 to 13 dpi. Antibody response against inoculated FAdV was appeared from 9 dpi. These results confirmed that the FAdV 11 strain is pathogenic in chicken. This study is the first experimental infection of FAdV 11 in chicken in Morocco, which increase our understanding of its pathogenicity in chickens and indicate that preventive measures against FAdV infection in poultry farms should be implemented in Morocco.

International audience; The depth of intravaginal insemination to achieve optimum fertility with frozen-thawed semen is highly species specific in birds and differ even in breed and/or strains of a species. Therefore, study was designed to evaluate the influence of intravaginal insemination depths (2 and 4 cm) on fertility outcome in Indian red jungle fowl. Semen collected from eight mature cocks was pooled, diluted in extender and cooled to 4 degrees C. Glycerol (20%) was added to chilled semen, equilibrated for 10 min and cryopreserved. After 3 days of storage, frozen semen was thawed in water bath at 37 degrees C for 30 s. After glycerol removal, intravaginal Inseminations were performed at the depth of 2 and 4 cm. The no. of fertilized eggs (31.4 +/- 1.6 vs. 27.7 +/- 1.8), fertility rate (65.7 +/- 3.6 vs. 58.8 +/- 4.0), no. of hatched chicks (27.8 +/- 1.9 vs. 23.5 +/- 1.6), hatchability of set eggs (58.8 +/- 4.3 vs. 49.7 +/- 3.2) and hatchability of fertilized eggs (88.4 +/- 2.8 vs. 84.3 +/- 2.2) were recorded higher with intravaginal depth of 4 cm compared to 2 cm. It is concluded that intravaginal insemination at the depth of 4 cm enhances the fertility outcomes of the frozen-thawed Indian red jungle fowl semen.

Hunters in Europe gather non-survey game species population estimates to inform wildlife management, however, the quality of such estimates remains unclear. We compared estimates of game density, realized annual intrinsic growth rates, and period mean growth rates between hunter obtained data and data obtained by targeted survey methods for four species in Poland from 1960 to 2014. Raw hunter estimates were strongly positively correlated to spotlight counts of red fox (18 years of monitoring), strip counts of brown hare (21 years) and grey partridge (25 years), male call counts of partridge (24 years), and complete counts of roe deer (49 years), and not related to spotlight counts of brown hare (15 years). Realized annual intrinsic growth rates derived from hunter estimates were strongly positively related to annual intrinsic growth rates derived from strip counts of grey partridge and complete counts of roe deer, but only weakly or not related to strip counts of brown hare, spotlight counts of red fox and brown hare, and male call counts of grey partridge. The period length at which the period mean growth rates derived from hunter estimates and estimates from other methods were strongly correlated was largely variable among methods and species. In the roe deer, correlation between these variables was strong across all years, while in smaller game species the period mean growth rates based on hunter estimates and other methods had the strongest association in period lengths of 6 to 11 years. We conclude that raw hunter estimates convey largely similar information to that provided by other targeted survey methods. Hunter estimates provide a source of population data for both the retrospective and prospective analysis of game population development when more robust estimates are unavailable.

The avian influenza virus H13 subtype circulates primarily in waterfowl. To explore the ability of the H13 virus to cross the host barriers, we genetically analysed two H13 isolates from wild birds in China and evaluated the infectivity of these subtypes in chickens. Genetic and molecular analyses showed differences in the lineages and amino acid sequences between the two subtypes; A/mallard/Dalian/DZ-137/2013 (H13N6) belonged to Group I, while A/Eurasian Curlew/Liaoning/ZH-385/2014 (H13N8) belonged to Group III. The nucleotide sequence results showed high homology (approximately 96.9%-100%) to sequences in GenBank. Neither H13 isolate replicated in adult chickens or 20-day-old chicks; however, the H13N8 strain replicated in 1- and 10-day-old chicks. Viruses were recovered from the nasal turbinate, tracheal, lung and colon tissues of chicks at 1, 3 and 5 days post-inoculation. The H13N6 isolates replicated inefficiently in 1-day-old chicks and did not replicate in 10-day-old chicks. Serological surveillance results showed that domestic chickens had a 4.6%-10.4% (15/328-34/328) positive antibody titre to the H13 virus. H13N6 and H13N8 isolates replicated in mammalian cell lines, including 293T, Madin-Darby canine kidney and chicken embryo fibroblast cells. Our results suggest that the AIV H13 subtype may cross the host barrier from wild waterfowl to land fowl.

Infectious bursal disease virus (IBDV), a double-stranded RNA virus, causes immunosuppression and high mortality in 3–6-week-old chickens. Innate immune defense is a physical barrier to restrict viral replication. After viral infection, the host shows crucial defense responses, such as stimulation of antiviral effectors to restrict viral replication. Here, we conducted RNA-seq in avian cells infected by IBDV and identified TRIM25 as a host restriction factor. Specifically, TRIM25 deficiency dramatically increased viral yields, whereas overexpression of TRIM25 significantly inhibited IBDV replication. Immunoprecipitation assays indicated that TRIM25 only interacted with VP3 among all viral proteins, mediating its K27-linked polyubiquitination and subsequent proteasomal degradation. Moreover, the Lys854 residue of VP3 was identified as the key target site for the ubiquitination catalyzed by TRIM25. The ubiquitination site destroyed enhanced the replication ability of IBDV in vitro and in vivo. These findings demonstrated that TRIM25 inhibited IBDV replication by specifically ubiquitinating and degrading the structural protein VP3., Author summary Owing to the immunosuppression and high mortality caused by infectious bursal disease virus (IBDV) in chickens, hosts urgently boost their antiviral immune responses to resist IBDV infection. Here, we identified the host antiviral factor TRIM25, which defended against IBDV infection by direct targeting the viral protein VP3 for subsequent ubiquitination and degradation. To our knowledge, this is the first report of host TRIM25 restricting IBDV replication. Additionally, we found that Lys854 of VP3 was the target site ubiquitinated by TRIM25. Mutation in the ubiquitination site of VP3 abrogated the degradation of VP3 and further enhanced IBDV replication, but did not influence its virulence. Our work not only uncovers an all-important molecular mechanism of chicken TRIM25 against IBDV but also provides new insights for developing high-titer live IBDV vaccines.

Fowl aviadenovirus (FAdV) species D and E are associated with inclusion body hepatitis (IBH); species C, serotype 4 (hereafter, FAdV4) is associated with hepatitis–hydropericardium syndrome (HHS) in young chickens. Outbreaks of HHS have led to significant losses in the poultry industry in several countries, predominantly in China. In April 2020, FAdV4 was detected in a remote backyard flock in California. In a mixed flock of chickens of various breeds and ages (6 mo to 2 y old), 7 of 30 were found dead within a week without premonitory signs. One additional bird died after the flock was relocated to fresh pasture, bringing the total mortality to 8 of 30 (27%). Postmortem examination of 3 birds revealed good body condition scores and active laying. One chicken had subtle hemorrhages throughout the liver, and the other 2 had diffusely dark mahogany livers. On histopathology, 2 chickens had hepatic necrosis with hepatocytes containing large, mostly basophilic, intranuclear inclusion bodies, identified by electron microscopy as 82.2-nm diameter adenoviral particles. Virus isolation and genomic sequencing performed on a liver sample revealed strains with 99.9% homology to FAdV4 isolates reported from China. To our knowledge, FAdV4 has not been reported in the United States to date. Furthermore, the chickens affected here were all adults and exhibited a variation of serotype 4 disease in which IBH was present but not hydropericardium.

Spermatogenesis is a complex cellular process that includes many subcellular events that are essential for the production of healthy spermatozoa. Autophagy is a physiological process that plays a significant role in the process of spermatogenesis; however, autophagy during avian spermatogenesis has not yet been reported. In the current study, we characterized in vivo autophagy throughout the process of domestic fowl spermatogenesis. Autophagy-specific markers, including microtubule-associated protein light chain 3 (LC3), sequestosome 1 (p62), and autophagy-related 7 (Atg7), were used to confirm the occurrence of autophagy in testicular germ cells. The protein expression of Atg7, LC3, and p62 in domestic fowl testes was confirmed by Western blotting. The immunohistochemical staining indicated a strong localization of LC3 and Atg7 within spermiogenic cells (intermediate and late spermatids) and primary spermatocytes. However, poorly expressed in cells (spermatogonia) that were located near the basement membrane. The immunofluorescence staining results showed the opposite tendency for LC3 and p62. LC3 was more strongly localized within the elongated spermatids, while p62 was strongly localized within the early spermatids. Moreover, the ultrastructural components of autophagy were revealed by transmission electron microscopy. Well-developed autophagosomes and multivesicular bodies were found to be prominent in primary spermatocytes (zygotene and pachytene) and spermiogenic cells. Furthermore, other vesicular structures, such as early endosomes and amphisomes, were also observed during spermatogenesis. The above findings collectively suggest that autophagy is active during spermatogenesis and that the level of autophagy increases from the basal to the luminal regions of the seminiferous tubules of domestic fowl testes. We propose that autophagic pathways may be involved in multiple functions to sustain spermatogenesis.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is considered to be one of the most important causes of foodborne diseases. Objective: The current examination was performed to examine the distribution of staphylococcal cassette chromosome mec (SCCmec) and Panton-Valentine leukocidin (PVL) gene amongst the MRSA strains isolated from raw fowl meat samples. Materials and Methods: A total of 240 fowl meat samples were collected and cultured. MRSA strains were identified using cefoxitin and oxacillin susceptibility tests. DNA samples extracted from the MRSA strains were subjected to polymerase chain reaction (PCR) for detection of SCCmec and PVL gene. Results: Twenty-two out of 240 (9.16%) raw fowl meat samples were positive for S. aureus strains. Twelve out of 22 S. aureus strains (54.54%) were determined as MRSA strains. The incidence of MRSA strains in raw chicken, turkey, quail, and ostrich meat samples was 66.66%, 50%, 50%, and 33.33%, respectively. The incidence of SCCmec IVa, SCCmec IVd, and SCCmec V was 50%, 8.33% and 41.66%, respectively. The applied method failed to detect SCCmec types I, II, III, IVb, and IVc. The incidence of the PVL gene amongst the MRSA strains was 75%. Conclusion: The presence of SCCmec IV and SCCmec V and PVL gene revealed occurrence of community-associated MRSA (CA-MRSA) in fowl meat samples. Further studies are required to find additional epidemiological aspects of the MRSA strains in fowl meat samples.

The poultry red mite (PRM) is one of the most economically important ectoparasites of laying hens globally. This mite can have significant deleterious effects on its fowl host including distress, anemia, reduced egg production, and reduced egg quality. This study was conducted to evaluate the influence of PRM on the serum protein profile in laying hens and its effect on the acute phase proteins (APPs) to assess their potential as biomarkers for mite infestation.\ud \ud Three APPs: alpha-1 acid glycoprotein (AGP), serum amyloid-A (SAA), and ceruloplasmin (CP) were measured in serum samples collected from laying hens at 12 and 17 wk of age, and then for up to 4 mo after a challenge with PRM (starting at 18.5 wk of age). The serum protein profile (SDS-PAGE/nanoflow HPLC electrospray tandem mass spectrometry) and concentration of individual serum proteins (SDS-PAGE-band densitometry) were also compared.\ud \ud Post challenge there was a positive correlation (r = 0.489; P < 0.004) between the levels of SAA and the PRM numbers. The levels of SAA steadily increased after the PRM challenge and were significantly different than the pre-challenge levels at 28, 32, and 36 wk of age (P < 0.01). The PRM numbers also peaked around 31-33 wk of age. The results for AGP and CP in comparison were inconsistent. Proteomics revealed the presence of 2 high molecular weight proteins in the serum between 12 and 17 wk of age. These were identified as Apolipoprotein-B and Vitellogenin-2, and their increase was commensurate with the onset of lay. No other major differences were detected in the protein profiles of blood sera collected pre and post challenge. We conclude that SAA could be used as a useful biomarker to monitor PRM infestation in commercial poultry flocks and that PRM infestation does not disrupt the production of the major proteins in the serum that are associated with egg formation.

Toxoplasmosis is a zoonotic diseases caused by Toxoplasma gondii ,which spread out by cats and it can be found in fowl. It because the fowl swallowing food which had been infected by Toxoplasma gondii oocyst. The most consumpted fowl is chicken.The source of data and agriculture information center showed that consumption of chicken was increased about 10,20%. It could be a potential increasement of Toxoplasma gondii infection. The objective of this research was indentificate the infection of Toxoplasma gondii cyst on ras chicken brain. The type of this research was a descriptive research, observe by examining 30 samples of the ras chicken brain by randomized sampling techniques. Based on research on 30 samples of the ras chicken brain, obtained positive results as much as 2 samples (6.6%) cyst infected of Toxoplasma gondii and others 28 samples of ras chicken brain (93.4%) were not infected with Toxoplasma gondii cysts. Positive results of Toxoplasma gondii can be caused due to maintenance system and cleanliness of chicken coop was much less, so there were carrier vectors contaminate to feed place, so there was a potential infection of Toxoplasmosis.It can be concluded that been 2 samples of chicken brain being infected cyst of Toxoplasma gondii or about 6,6% and there were 28 samples or 93,4% of chicken brain that were not infected by cyst of Toxoplasma gondii . Infection circle of Toxoplasmosis can be avoided by manage maintenance system of chicken and keep the cage cleamliness, also keep the personal hygiene and feeds.

Inclusion body hepatitis (IBH) is, in some cases, a fatal disease affecting fowl by adenovirus strains which are subdivided into 5 species (A-E). In the current study, we investigated sequences from the Loop L1 region of the hexon gene of sequences of adenovirus field stains 1/A and 11/D isolated from a poultry flock co-infected with IBH and avian reoviruses ARVs. In early 2021, an epidemiologic survey highlighted the coinfection adenoviruses with other viruses (orthoreovirus infection) as being particularly deleterious within the poultry industry. Here, we investigated the Loop L1 HVR1-4 region of the hexon gene with relative synonymous codon usage (RSCU) designation and RSCU inclusive of all the mutations. These are the first results that have been presented on fowl adenovirus species A and D with simultaneous reovirus infection in 38-days old broiler chickens in Poland.

Fowl adenoviruses (FAdV), detected during routine diagnostic investigations from 38 countries (5 continents) over a decade, were partially sequenced and grouped by phylogenetic analysis. The partial polymerase gene nucleotide sequences of the 365 fowl adenovirus isolates resulted in the following species distribution: 11% FAdV-A; 3% FAdV-B; 2% FAdV-C; 34% FAdV-D; and 50% FAdV-E. Noticeably, only 79 of the detected strains could be associated with adenovirus-specific pathologic conditions: 62 (79%) with inclusion body hepatitis; 9 (11%) with gizzard erosion; and 8 (10%) with hepatitis hydropericardium syndrome. The remainder of the FAdV strains was detected as concomitant infection from other disease conditions almost exclusively in boilers of 27 to 42 d of age: the majority of them was FAdV-E followed by FAdV-D, and to a lesser extent of FAdV-A, B, and C, the latter ones have not been associated with any of the established adenovirus-caused syndromes in our collection. The highest ratio of coinfections was observed for FAdV-B (62%), while it was about 30% for the rest of the FAdV species. The most frequent coinfection, in connection with all FAdV species, was with the avian infectious bronchitis virus. The presented database will serve as the basis for comparative whole genome and cross-neutralization analysis of selected FAdV isolates.

Circular RNAs (circRNAs) play important roles in adipogenesis. However, studies on circRNA expression profiles associated with the development of abdominal adipose tissue are lacking in chickens. In this study, 12 cDNA libraries were constructed from the abdominal adipose tissue of Chinese domestic Gushi chickens at 6, 14, 22, and 30 weeks. A total of 1,766 circRNAs were identified by Illumina HiSeq 2500 sequencing. These circRNAs were primarily distributed on chr1 through chr10 and sex chromosomes, and 84.95% of the circRNAs were from gene exons. Bioinformatic analysis showed that each circRNA has 35 miRNA binding sites on average, and 62.71% have internal ribosome entry site (IRES) elements. Meanwhile, these circRNAs were primarily concentrated in TPM < 0.1 and TPM > 60, and their numbers accounted for 18.90% and 80.51%, respectively, exhibiting specific expression patterns in chicken abdominal adipose tissue. In addition, 275 differentially expressed (DE) circRNAs were identified by comparison analysis. Functional enrichment analysis showed that the parental genes of DE circRNAs were primarily involved in biological processes and pathways related to lipid metabolism, such as regulation of fat cell differentiation, fatty acid homeostasis, and triglyceride homeostasis, as well as fatty acid biosynthesis, fatty acid metabolism, and glycerolipid metabolism. Furthermore, ceRNA regulatory networks related to abdominal adipose development were constructed. The results of this study indicated that circRNAs can regulate lipid metabolism, adipocyte proliferation and differentiation, and cell junctions during abdominal adipose tissue development in chickens through complex ceRNA networks between circRNAs, miRNAs, genes, and pathways. The results of this study may help to expand the number of known circRNAs in abdominal adipose tissue and provide a valuable resource for further research on the function of circRNAs in chicken abdominal adipose tissue.

Intensification of agricultural practices has drastically shaped farmland landscapes and generally caused a decline in spatial and temporal heterogeneity, thus leading to changes in habitat quality and food resources and a decline for most farmland birds Europe-wide. The relationship between complex landscape changes and habitat preferences of animals still remains poorly understood. Particularly, temporal and spatial changes in diversity may affect not only habitat choice but also population sizes. To answer that question, we have looked into a severely declining typical farmland bird species, the grey partridge Perdix perdix in a diverse farmland landscape near Vienna to investigate the specific habitat preferences in respect to the change of agricultural landscape over two decades and geographic scales. Using a dataset collected over 7.64 km² and between 2001 and 2017 around Vienna, we calculated Chesson’s electivity index to study the partridge’s change of habitat selection over time on two scales and between winter and spring in 2017. Although the farmland landscape underwent an ongoing diversification over the two decades, the grey partridges declined in numbers and shifted habitat use to less diverse habitats. During covey period in winter, partridges preferred also human infrastructure reservoirs such as roads and used more diverse areas with smaller fields than during breeding where they selected harvested fields but surprisingly, avoided hedges, fallow land and greening. Known as best partridge habitats, those structures when inappropriately managed might rather function as predator reservoirs. The avoidance behaviour may further be a consequence of increasing landscape structuring and edge effects by civilisation constructions. Besides, the loss in size and quality of partridge farmland is altered by crop choice and pesticides reducing plant and insect food. With declining breeding pairs, the grey partridge does not seem to adjust to these unsustainable landscape changes and farmland practices.

Microvascularization of domestic fowl kidneys was studied using scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Two types of nephrons, mammalian-type (MT) and reptilian-type (RT) nephrons and their glomerular structure were analysed quantitatively by 3D morphometry. A significant difference in shape and size between the MT and RT glomeruli was found. The mean diameter of the RT glomeruli was about 56 µm, while that of MT glomeruli was significantly larger, namely about 80 µm. The afferent arterioles in mammalian-type glomeruli usually bifurcated into two lobular branches and formed a complex glomerular capillary network with numerous loops. Reptilian-type glomeruli consisted of a single capillary forming few loops and leaving the glomerulus as efferent arteriole. Diameters of afferent and efferent arteriolar replicas were similar in all three kidney divisions of MT and RT nephrons. The absence of the interconnecting branches between the MT nephron capillaries at the gross inspection suggests that the mammalian-type nephron glomeruli, although more complex than the reptilian type, are not equivalent to those in mammalian kidneys.

Avian hepatitis E virus (aHEV) is associated with hepatitis-splenomegaly syndrome, big liver and spleen disease and hepatic rupture haemorrhage syndrome. However, the knowledge about aHEV in commercial layer chickens in Nigeria is scarce. In this study, 460 serum samples obtained from 36 apparently healthy commercial layer chicken flocks in three states (Ogun, Osun and Oyo States) of southwestern Nigeria were analysed by enzyme linked immunosorbent assay for the presence of anti-aHEV immunoglobulin Y (IgY) antibodies. In total, the overall seroprevalence of anti-aHEV antibodies was 14.6%. The serological analysis revealed that 75% of the flocks examined were positive for anti-aHEV IgY antibodies from chickens of various ages in all three states. The percentage of the seropositive chickens in the three states varied from flock to flock ranging from 60% to 88.8% and seropositive chickens were detected at any age (24–52 weeks of age) without significant differences between the age groups. This is the first report assessing the presence of aHEV antibodies in chickens from Nigeria. The detection of anti-aHEV antibodies in commercial layer chickens in this study emphasizes the importance of serosurveillance in disease monitoring due to the economic threat posed by aHEV as a result of decreased egg production and increased mortality in affected commercial layer chicken farms. However, further studies are essential to reveal the clinical implications and to assess the real burden of aHEV in Nigeria.

Telocytes (TCs), a novel type of interstitial cells, were identified in various animals. Since TCs have not observed in avian skin, hence, we carried out immunohistochemistrical and transmission electron microscopical studies in the skin of the silky fowl to investigate the TCs. TCs appear as CD34, c-Kit, and PDGFRα immunopositive. The elongated TCs with 2 long and thin telopodes (Tps) are located in the dermis. Generally, a TC possesses a fusiform, ovoid and polygonal cell body with 2 Tps (lengths = 5.27–21.85 μm), which are uneven in thickness including thick sections – podoms (diameters = 0.40–0.47 μm) and thin sections – podomers (diameters = 0.03–0.04 μm). TCs/Tps are observed frequently in close proximity to neighboring cell types/structures, such as adipocytes, collagen fibers, and capillaries. Under a magnified field, homocellular TCs/Tps contacts are observed through gap junctions (distances = 0.01–0.05 μm), whereas some of TCs/Tps have heterocellular close contacts by point contacts with surrounding cells, including stem cells and melanocytes. The multivisicular bodies, especially exosomes (diameters = 0.09–0.23 μm) releasing from TCs/Tps are observed in close proximity to TCs/Tps. Our results illustrated that the novel type of interstitial cells – TCs are present in the dermis of the silky fowl, and they have special structural relationships with surrounding cell types. The study provides histological evidence for TCs involvement in intercellular communication, skin regeneration, and pigmentogenesis in avian skin.

Arcobacter butzleri is an emerging zoonotic food-borne and water-borne pathogen that can cause diarrhea in humans. The global prevalence of A. butzleri infection is underestimated, and little is known about their phenotypic and genotypic characterization. The aim of this study was to determine antimicrobial susceptibility (AST) profiles, detect related virulence genes, and classify sequence type (ST) of A. butzleri isolates obtained from human stool and food samples. A total of 84 A. butzleri isolates were obtained from human diarrheal (n = 25), non-diarrheal (n = 24) stool, and food (n = 35) samples in Thailand. They were evaluated for phenotypic identification by conventional microbiological procedures and AST by Kirby-Bauer disc diffusion method as well as virulence genes detection. Representative isolates from each origin were selected based on the presence of virulence genes and AST profiles to analyze genetic diversity by multilocus sequence typing (MLST). All isolates showed resistance to nalidixic acid 40.5% (34/84), ciprofloxacin 11.9% (10/84), azithromycin 8.3% (7/84), and erythromycin 3.6% (3/84). Regarding the ten virulence genes detected, cj1349, mviN and pldA had the highest prevalence 100% (84/84), followed by tlyA 98.8% (83/84), cadF 97.6% (82/84), ciaB 71.4% (60/84), hecA and hecB 22.6% (19/84), iroE 15.5% (13/84) and irgA 10.7% (9/84), respectively. Three virulence genes were present among A. butzleri isolates of human diarrheal stool and food samples, with a significant difference observed among isolates; hecB [36% (9/25) and 8.6% (3/35)], hecA [36% (9/25) and 5.7% (2/35)], and irgA [24% (6/25) and 2.9% (1/35)] (p < 0.05), respectively. The hecA and hecB virulence genes functions are related to the mechanism of hemolysis, while irgA supports a bacterial nutritional requirement. MLST analysis of 26 A. butzleri isolates revealed that 16 novel STs exhibited high genetic diversity. The results of this study is useful for understanding potentially pathogenic and antimicrobial-resistant A. butzleri in Thailand. The pathogenic virulence markers hecB, hecA, and irgA have the potential to be developed for rapid diagnostic detection in human diarrheal stool. No significant relationships among STs and sources of origin were observed. Little is known about A. butzleri, the mechanism of action of these virulence genes, is a topic that needs further investigation.

Domestic ducks are of paramount importance as a cheap source of protein in rural India. Andaman local duck (ALD) is an indigenous avian genetic resource of Andaman and Nicobar islands (ANI) and is mainly distributed in Middle and Northern parts of these islands. Negligence has brought this breed on the edge of extinction necessitating immediate conservation efforts. Here, we report the genetic diversity, population structure and matrilineal genetic root of ALD. Partial mtDNA D-loop sequences were analyzed in 71 ALD samples and analysis revealed 19 polymorphic sites and 13 haplotypes. Estimated haplotype (Hd ± SD) and nucleotide diversity (π ± SD) were 0.881 ± 0.017 and 0.00897 ± 0.00078 respectively. The high genetic diversity of ALD indicates introgression of genetic material from other local duck breeds. In addition, it can be postulated that ALD bearing high genetic diversity has strong ability to adapt to environmental changes and can withstand impending climate change. Phylogenetic and network analysis indicate that ALD falls under Eurasian clade of mallard and ALD forms three clusters; one cluster is phylogenetically close to Southeast Asian countries, one close to Southern part of mainland India and the third one forms an independent cluster. Therefore, ALD might have migrated either from Southeast Asian countries which enjoy a close cultural bondage with ANI from time immemorial or from Southern part of India. The independent cluster may have evolved locally in these islands and natural selection pressure imposed by environmental conditions might be the driving force for evaluation of these duck haplotypes; which mimics Darwin’s theory of natural selection. The results of the study will be beneficial for formulating future breeding programme and conservation strategy towards sustainable development of the duck breed.

Repeated outbreaks due to H3N1 low pathogenicity avian influenza viruses (LPAIV) in Belgium were associated with unusually high mortality in chicken in 2019. Those events caused considerable economic losses and prompted restriction measures normally implemented for eradicating high pathogenicity avian influenza viruses (HPAIV). Initial pathology investigations and infection studies suggested this virus to be able to replicate systemically, being very atypical for H3 LPAIV. Here, we investigate the pathogenesis of this H3N1 virus and propose a mechanism explaining its unusual systemic replication capability. By intravenous and intracerebral inoculation in chicken, we demonstrate systemic spread of this virus, extending to the central nervous system. Endoproteolytic viral hemagglutinin (HA) protein activation by either tissue-restricted serine peptidases or ubiquitous subtilisin-like proteases is the functional hallmark distinguishing (H5 or H7) LPAIV from HPAIV. However, luciferase reporter assays show that HA cleavage in case of the H3N1 strain in contrast to the HPAIV is not processed by intracellular proteases. Yet the H3N1 virus replicates efficiently in cell culture without trypsin, unlike LPAIVs. Moreover, this trypsin-independent virus replication is inhibited by 6-aminohexanoic acid, a plasmin inhibitor. Correspondingly, in silico analysis indicates that plasminogen is recruitable by the viral neuraminidase for proteolytic activation due to the loss of a strongly conserved N-glycosylation site at position 130. This mutation was shown responsible for plasminogen recruitment and neurovirulence of the mouse brain-passaged laboratory strain A/WSN/33 (H1N1). In conclusion, our findings provide good evidence in natural chicken strains for N1 neuraminidase-operated recruitment of plasminogen, enabling systemic replication leading to an unusual high pathogenicity phenotype. Such a gain of function in naturally occurring AIVs representing an established human influenza HA-subtype raises concerns over potential zoonotic threats., Author summary Avian influenza viruses (AIV) of subtype H3N1 caused outbreaks among gallinaceous poultry associated with an unusual high mortality in Belgium in 2019. Although those strains shared properties of a high pathogenicity (HP)-like phenotype in chicken experiments, still the viral hemagglutinin endoproteolytic cleavage site (HACS) matches that of canonical low pathogenicity (LP) AIV. Above all, the HACS motif is the molecular marker to distinguish low from high pathogenicity phenotypes in H5 and H7 AIV. Monobasic HACS motifs restrict the LPAIV to use tissue-specific host proteases and therefore confine their organ tropism to the respiratory and digestive tracts. By contrast, the polybasic HACS of HPAIV provides access to the ubiquitously expressed prohormone convertase furin, resulting in a grossly broadened organ tropism. Here, we show that the systemic spread of the Belgian H3N1 chicken viruses is independent from tissue-specific proteases and that their proteolytic HA activation is processed by plasminogen and associated to the loss of a conserved N-glycosylation site at position 130 of the neuraminidase. To date, this mechanism was only described for the mouse-adapted laboratory H1N1 strain WSN/33. Our data, however, provide evidence that this mechanism can also evolve in natural circulating AIV, as well as in human strains, suggesting a certain zoonotic potential.

Background Poultry represent a widely held economic, nutritional, and sociocultural asset in rural communities worldwide. In a recent longitudinal study in western Kenya, the reported mean number of chickens per household was 10, with increases in flock size constrained principally by mortality. Newcastle disease virus is a major cause of chicken mortality globally and hypothesized to be responsible for a large part of mortality in smallholder flocks. Our goal was to determine the impact of routine Newcastle disease virus (NDV) vaccination on flock size and use this data to guide programs to improve small flock productivity. Methods We conducted a factorial randomized controlled trial in 537 households: in 254 households all chickens were vaccinated every 3 months with I-2 NDV vaccine while chickens in 283 households served as unvaccinated controls. In both arms of the trial, all chickens were treated with endo- and ecto parasiticides every 3 months. Data on household chicken numbers and reported gains and losses were collected monthly for 18 months. Results Consistent with prior studies, the overall flock size was small but with increases in both arms of the study over time. The mean number of chickens owned at monthly census was 13.06±0.29 in the vaccinated households versus 12.06±0.20 in the control households (p = 0.0026) with significant gains in number of chicks (p = 0.06), growers (p = 0.09), and adults (p = 0.03) in the vaccinated flocks versus the controls. Household reported gains were 4.50±0.12 total chickens per month when vaccinated versus 4.15±0.11 in the non-vaccinated controls (p = 0.03). Gains were balanced by voluntary decreases, reflecting household decision-making for sales or household consumption, which were marginally higher, but not statistically significant, in vaccinated households and by involuntary losses, including mortality and loss due to predation, which were marginally higher in control households. Conclusion Quarterly NDV vaccination and parasiticidal treatment resulted in an increase in flock size by a mean of one bird per household as compared to households where the flock received only parasiticidal treatment. While results suggest that the preventable fraction of mortality attributable to Newcastle disease is comparatively small relatively to all-cause mortality in smallholder households, there was a significant benefit to vaccination in terms of flock size. Comparison with previous flock sizes in the study households indicate a more significant benefit from the combined vaccination and parasiticidal treatment, supporting a comprehensive approach to improving flock health and improving household benefits of production in the smallholder setting.

Fowl adenoviruses (FAdV) are important infectious pathogens responsible for causing substantial economic losses to the poultry industry worldwide. One hundred and forty-six FAdV strains were continuously collected and analysed from 2013 to 2019 to understand the epidemiological change and nature of the virus in South Korea from two different standpoints, before and after the release of multiple commercial FAdV-4 vaccines. Phylogenetic analysis of the hexon loop-1 gene sequences showed that 92 strains belonged to FAdV-C (63%), 35 strains to FAdV-E (24%), 18 strains to FAdV-D (12.3%), and one strain to FAdV-A (0.7%), respectively. We provide evidence that the dominant FAdV serotype has recently changed from FAdV-4 to FAdV-8b, as reflected in the proportion of each serotype in field cases in 2019 (18.5% and 77.8%, respectively). The newly emerged FAdV-8b cluster was significantly noticeable compared to the old FAdV clusters, indicating that the development of a vaccine for FAdV-8b may be necessary. Overall, this new insight into FAdV prevalence provides a foundation for strategic control and the development of efficient vaccines against FAdV cases in chickens in South Korea.

Pen enrichment for broiler (meat-type) chickens is one of the potential strategies to stimulate locomotion and consequently contribute to leg health and welfare. This study was designed to evaluate effects of using a plethora of pen enrichments (barrier perches, angular ramps, horizontal platforms, large distance between feed and water and providing live Black Soldier fly larvae in a dustbathing area) on tibia characteristics, locomotion, leg health and home pen behaviour of fast and slower-growing broiler chickens. The experiment was set up as a 2 x 2 factorial arrangement with a total of 840 male broiler chickens in a complete randomized design (7 replicates per treatment and 30 chickens per replicate) with the following treatments: 1) pen enrichment (enriched pen or non-enriched pen); 2) broiler strain (fast-growing Ross 308 or slower-growing Hubbard JA 757). Home pen behaviour and use of enrichment were observed. At approximately 1400 and 2200 gram body weight, two chickens per pen were randomly selected and slaughtered, to investigate tibia morphological, biophysical and mechanical characteristics and leg health. Pen enrichment positively affected tibia biophysical characteristics, e.g., osseous volume (Δ=1.8 cm3, P=0.003), total volume (Δ=1.4 cm3, P=0.03) and volume fraction (Δ=0.02 %, P=0.002), in both fast and slower-growing chickens, suggesting that pen enrichment particularly affects ossification and mineralization mechanisms. Accordingly, locomotion and active behaviours were positively influenced by pen enrichment. However, pen enrichment resulted in lower body weight gain in both strains, which might be due to higher activity or lower feed intake as a result of difficulties of crossing the barrier perches. Regarding the strain, slower-growing chickens showed consistently more advanced tibia characteristics and more active behaviour than fast-growing chickens. It can be concluded that pen enrichment may lead to more activity and better bone development in both fast and slower-growing chickens.

The Eastern Pamir (eastern Tajikistan) is a high-mountain plateau with elevations up to 7000 m, currently characterized by extremely severe environmental conditions and harboring a specialized montane fauna, which in part is shared with that of the Tibetan Plateau. The modern bird fauna of High Asia comprises a diversity of both ancient and recently diverged endemics, and thus is of general importance for historical biogeography and understanding the origin of modern high mountain ecosystems. However, the past history of the Central Asian highland avian communities remains practically unknown, as no fossil bird assemblages from high elevation areas were previously reported. In particular, it remains completely unexplored how birds responded to late Quaternary climatic fluctuations. Here we report the first fossil bird fauna from the High Asia and the first fossil birds from Tajikistan. An assemblage from the late Pleistocene through middle Holocene of Istykskaya cave (4060 m) in Eastern Pamir surprisingly comprises a remarkable diversity of waterbirds, including a few species that are completely absent from High Asia today and that were not reported globally from such high altitudes. The diversity of waterbirds incudes taxa of various ecological preferences (shorebirds, underwater and surface feeders, both zoophagous and phytophagous), strongly indicating the presence of a productive waterbody at the vicinity of the site in the past. These observations correspond to recent palaeoclimatic data, implying increased water availability in this region, currently occupied by high mountain semi-deserts. Our findings for the first time show that milder environmental conditions of late Quaternary attracted lowland species to the Central Asian highland wetlands. The reported assemblage yet contains several characteristic highland taxa, indicating a long-time persistence of some Central Asian montane faunistic elements. In particular, it includes the Tibetan Sandgrouse (Syrrhaptes tibetanus), a highly-specialized montane dweller, which is for the first time found in the fossil record.

Salmonella is a zoonotic pathogen that persists in poultry. Salmonella vaccines that can be delivered in-ovo can be cost-effective and can decrease Salmonella load in poultry. This study evaluates the efficacy of a Salmonella chitosan-nanoparticle (CNP) vaccine, administered in-ovo, in broilers. CNP vaccine was synthesized with Salmonella Enteritidis (SE) outer-membrane-proteins (OMPs) and flagellin proteins. At embryonic-d18, one-hundred-thirty-six eggs were injected with 200μl PBS or 1000μg CNP into the amniotic cavity. At d1-of-age, 132 chicks were allocated in 6 pens/treatment with 11 chicks/pen. At d7, birds were orally challenged with 1×109 CFU/bird SE. At d1, 8h-post-challenge, d14, and d21, serum anti-SE-OMPs IgY were analyzed. At d14 and d21, cloacal swabs and bile anti-SE-OMPs IgA, CD4+/CD8+-T-cell ratios, and ceca SE loads were analyzed. At d21, cecal tonsil IL-1β, IL-10, and iNOS mRNA were analyzed. Body-weight-gain (BWG) and feed-conversion-ratio (FCR) were recorded weekly. Data were analyzed by Student’s t-test at P+/CD8+-T-cell ratios between vaccinated birds compared to control. There were no significant differences in IL-1β, IL-10, iNOS mRNA between vaccinated birds compared to control. Findings demonstrate that the in-ovo administration of CNP vaccine can induce an antigen-specific immune response against SE and can decrease SE cecal load in broilers.