Chicken Gene Research Articles

Polymorphism of ghrelin genes among four Nigerian chicken populations as tool for improvement of chickens

Research on polymorphism of ghrelin (GHRL), an acylated peptide that stimulates the release of growth hormone from the pituitary, has shown that it is associated with some essential growth genes in chicken, hence veritable in genomic selection. There is porous information on the polymorphism of GHRL genes among Nigerian chickens. The objective of this study was to determine polymorphism of ghrelin gene among four Nigerian chicken populations. Blood sample (0.5 ml) was collected from the wing vein of 102 birds for DNA extraction. Tested PCR products were sequenced following Macrogen INC. The SNPs were determined using the sequence alignment program, CLUSTAL W, implemented in MEGA software. The sequence results showed 25 SNPs. The Nigerian chickens varied in polymorphic sites from Ogun chickens (348 and 535) to Nsukka chickens (558 and 696) at the GHRL 1 and GHRL2 loci respectively. Polymorphic sites and diversity were higher among the Nsukka chicken than other populations of the Nigerian chickens. SNPs common to one geographic varied in another. The result revealed that genomic selection based on ghrelin SNPs may yield higher predictive accuracy, while Nsukka chickens could be useful for the creation of more superior lines for the global poultry industry. Since past studies have revealed that ghrelin gene stimulates the pituitary and hypothalamus for releasing growth factor (GF) hormone and have strong effect on many organs, we suggest an association study of ghrelin gene and growth traits.

Metagenomic Insights into Chicken Gut Antibiotic Resistomes and Microbiomes.

ABSTRACTThe chicken gut microbiota, as a reservoir of antibiotic resistance genes (ARGs), poses a high risk to humans and animals worldwide. Yet a comprehensive exploration of the chicken gut antibiotic resistomes remains incomplete. In this study, we established the largest chicken gut resistance gene catalogue to date through metagenomic analysis of 629 chicken gut samples. We found significantly higher abundance of ARGs in the Chinese chicken gut than that in the Europe. tetX, mcr, and blaNDM, the genes resistant to antibiotics of last resort for human and animal health, were detected in the Chinese chicken gut. The abundance of ARGs was linearly correlated with that of mobile genetic elements (MGEs). The host-tracking analysis identified Escherichia, Enterococcus, Staphylococcus, Klebsiella, and Lactobacillus as the major ARG hosts. Especially, Lactobacillus, an intestinal probiotic, carried multiple drug resistance genes, and was proportional to ISLhe63, highlighting its potential risk in agricultural production processes. We first established a reference gene catalogue of chicken gut antibiotic resistomes. Our study helps to improve the knowledge and understanding of chicken antibiotic resistomes for knowledge-based sustainable chicken meat production.IMPORTANCE The prevalence of antibiotic resistance genes in the chicken gut environment poses a serious threat to human health; however, we lack a comprehensive exploration of antibiotic resistomes and microbiomes in the chicken gut environment. The results of this study demonstrate the diversity and abundance of antibiotic resistance genes and flora in the chicken gut environment and identify a variety of potential hosts carrying antibiotic resistance genes. Further analysis showed that mobile genetic elements were linearly correlated with antibiotic resistance genes abundance, implying that we should pay attention to the role played by mobile genetic elements in antibiotic resistance genes transmission. We established a reference genome of gut antibiotic resistance genes in chickens, which will help to rationalize the use of drugs in poultry farming.

Study of Co1 and BIK BCL2 Gene Analysis in Gorontalo Local Chicken

Analysis studies of Co1 and BIK BCL2 genes in local Gorontalo chickens were carried out using real-time PCR. This study aims to detect the presence of Co1 and BIK BCL2 genes in local Gorontalo chickens so that they can be used as information on gene diversity that can be used as genetic markers to study the diversity of local chicken species in Indonesia. The method used in this study is the SYBR green real-time PCR method. The research conducted found that the concentration values of the extracted samples were in the range of 68.15-68.30 ng/µL. Meanwhile, the purity values measured at wavelength A260/A280 were obtained with purity ranges between 1.867-1.923. The amplification results of the Co1 target gene show that the Ct value is in the range of 20.90-21.20, while for Tm, it is 84.10-84.20. In the BIK BCL2 target gene analysis, the Ct value was in the range of 16.15-16.20, while for Tm, it was 91.10-91.20. From these results, it can be concluded that all research samples were sampled from six different regions detected with CT and Tm values from the real-time analysis carried out obtained homogeneous values and no more than 2 degrees of difference between samples. The research novelty was using Nisi chicken as the native, one of the local chickens in Indonesia as the object of study. Besides, we used a BIK BCL 2 gene as the marker to determine the genetic diversity of the local chicken. A suggestion for further research is to explore more diverse genes to obtain more data on genetic diversity.

Black Pepper or Radish Seed Oils in a New Combination of Essential Oils Modulated Broiler Chickens' Performance and Expression of Digestive Enzymes, Lipogenesis, Immunity, and Autophagy-Related Genes.

Optimal combinations of essential oils (EOs) can enhance performance and maintain poultry productivity. The effects of EOs with black pepper oil (BPO) or radish seed oil (RSO) on performance and the expression of digestive enzymes, lipogenesis, immunity, and autophagy-related genes in broiler chickens were explored. Six dietary treatments for 300 one-day-old chicks were allocated as follows: controls were fed a basal diet, one group was fed an EO-supplemented diet (1.5 g/kg diet of parsley, mint, and carrot seed oils (1:1:1)), and other groups received Eos + BPO0.25, Eos + BPO0.5, Eos + RSO0.25, and Eos + RSO0.5 treatments, with a basal diet containing EOs plus BPO or RSO at the level of 0.25 or 0.5 g/kg, respectively. Supplementation with 0.5 g/kg of EOs plus BPO or RSO resulted in the most improved maximum BWG and FCR in broiler chickens. The lactobacilli population was increased in Eos + BPO0.5, followed by EOs + RSO0.5, unlike in the control. The highest expression of the CCK and PNLIP genes was identified in the Eos + BPO group. The FAS and ACC genes were upregulated, while the IgA and IL-10 genes were downregulated, with EOs plus RSO or BPO. The group that received Eos + BPO0.5, followed by Eos + RSO0.5, displayed patterns of higher expression for atg5, atg7, and atg12, with lower expression of mTOR. In summary, a new combination of EOs with 0.5 g/kg BPO had potential growth-promoting and immune-boosting effects in broiler chickens.

Effects of dietary palm oil on broiler chicken productive performance and carcass characteristics: a comprehensive review

Palm oil is a natural energy source ingredient in poultry diets that offers a broad range of beneficial effects on the performance of broiler chickens. This review was conducted to highlight the impact of palm oil as a feed ingredient on growth performance and carcass quality, as well as the biochemical, antioxidant activity and tissue fatty acids (FA) composition of broiler chickens. Palm oil inclusion in broiler chickens' rations contributes significantly to the high metabolisable energy (ME) of feed formulation, increases feed palatability and decreases digesta passage rate in the intestine. The reviewed literature indicated that dietary palm oil has a beneficial effect on broiler chickens' overall growth performance traits. The addition of palm oil can also improve the heat tolerance of chickens reared in high ambient temperature conditions. Regardless of breed and breeding conditions, palm oil exhibits good oxidative stability in broiler chickens due to the presence of prevalent phytonutrient elements in this oil. The inclusion of palm oil increased palmitic (C16:0) and oleic (C18:1) acids in tissue deposits, which improves meat stability and quality. Moreover, molecular studies have revealed that higher mRNA expression of several lipid-related hepatic genes in broiler chickens fed palm oil. Nonetheless, dietary palm oil can influence FA deposition in tissues, modulate lipoprotein and triglycerides (TG) levels, and cytokine contents in the blood serum of broiler chickens.

Antimicrobial activity of sophorolipids against Eimeria maxima and Clostridium perfringens, and their effect on growth performance and gut health in necrotic enteritis

The in vitro antimicrobial activity of sophorolipids (SLs) against Eimeria maxima and Clostridium perfringens, and the in vivo effects of SLs on growth performance and gut health in necrotic enteritis (NE)-afflicted broiler chickens were studied. To test the direct killing effects of SLs on enteric pathogens, 2.5 × 105 freshly prepared sporozoites of each Eimeria acervulina, E. maxima, and E. tenella were placed in each well of a 96-well plate, and the vegetative stage of Clostridium perfringens was prepared at 1 × 109 cfu/well. Four different SLs (C18:1 lactonic diacetyled SL [SL1], C18:1 deacetyled SL [SL2], C18:1 monoacetyled SL [SL3], and C18:1 diacetyled SL [SL4]), and 2 anticoccidial chemical controls, decoquinate and monensin, were evaluated at 3 dose levels (125 µg/mL, 250 µg/mL, and 500 µg/mL). Samples were incubated at 41°C for 3 h, and microbial survival ratios were measured by using a cell counter to quantify the number of live microbes stained by fluorescent dye. A total of 336 (0-day-old) male commercial broiler chickens were used to assess the effects of SLs in vivo. Chickens were randomly allocated to 6 treatment groups (7 chickens per cage, 8 cages per treatment) as follows: a control group which received a basal diet (CON), a negative control group (NC) which received a basal diet and NE challenge, and 4 SL treatment groups with NE (NC+SL1, NC+SL2, NC+SL3, and NC+SL4). The inclusion rates of SLs in each group were 200 mg/kg of feed. NE-induced chickens were orally infected with E. maxima (10,000 oocysts/chicken) on d 14, followed by C. perfringens (1 × 109 cfu/chicken) on d 19. Disease parameters measured included gut lesion scores, intestinal cytokine production, and level of tight junction protein expression. Data were analyzed using a Mixed Model (PROC MIXED) in SAS. In vitro (Experiment 1), all SLs dose-dependently decreased (P < 0.001) the viability of the three species of Eimeria sporozoites and C. perfringens. In vivo (Experiment 2), dietary SLs increased (P < 0.001) body weight and average daily gain of broiler chickens infected with NE. Dietary SL1 and SL4s increased (P < 0.05) feed conversion ratio compared to NC. Furthermore, SL1 and SL4 decreased (P < 0.05) gut lesion scores in combination with increased expression of IL1β, IL8, TNFSF15, and IL10 genes (P < 0.05) in NE-afflicted chickens. Overall, dietary SLs promoted growth performance, intestinal immune responses, and intestinal barrier integrity of NE-afflicted, young broiler chickens.

Molecular cloning and characterization of partial promoter of Acetyl coenzyme A carboxylase B gene in chicken

Molecular cloning and characterization of partial promoter of Acetyl coenzyme A carboxylase B gene in chicken

Molecular genetic diversity and bioinformatic analysis of Leucocytozoon sabrazesi based on the mitochondrial genes cytb, coxI and coxIII and co-infection of Plasmodium spp.

Leucocytozoon sabrazesi is an intracellular haemoprotozoan parasite responsible for leucocytozoonosis, which is transmitted by insect vectors and affects chickens in tropical and subtropical areas in many countries. It causes huge economic losses due to decreased meat and egg production. In the present study, we used nested PCR to determine the genetic diversity of L. sabrazesi based on the cytb, coxI, coxIII and concatenated genes in chickens in Thailand. In addition, we found co-infections between L. sabrazesi and Plasmodium spp. (P. gallinaceum or P. juxtanucleare) in chickens that were not identified by microscopic examination of blood smears. The phylogenetic analysis indicated that L. sabrazesi cytb and coxIII genes were conserved with similarity ranging from 99.9 to 100% and 98 to 100%, respectively whereas the coxI gene was diverse, with similarities ranging from 97 to 100%. These findings ascertained the nucleotide analysis of the cytb, coxI, coxIII and concatenated sequences in which 4, 8, 10 and 9 haplotypes were found, respectively. In addition, it was found that the large number of synonymous substitutions and conservative amino acid replacements in these mitochondrial genes occurred by non-synonymous substitution. The evolutionary analysis of the Ka/Ks ratio supported purifying selection and the negative values of both Fu’s Fs and Tajima’s D indicate selective sweep especially for the coxI gene. The entropy and Simplot analysis showed that the genetic variation in populations of Plasmodium spp. was higher than in Leucocytozoon. Hence, the nucleotide sequences of three mitochondrial genes could reflect the evolutionary analysis and geographic distribution of this protozoan population that switches hosts during its life cycle.

Influence of Nano-Methionine supplementation in drinking water on growth performance, lipid metabolism, and related gene expression in broiler chicken.

The study aimed to determine how Nano-Methionine (Nano-Meth) affected growth, lipid metabolism, and relative gene expression for acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), growth hormone receptor (GHR), insulin-like growth factor receptor-1 (IGFR-1), myostatin (MSTN), and cholecystokinin (CCK) genes in broiler chickens. A total of 100 1-day-old broilers were randomly assigned into 2 groups: 1) the control group received drinking water without any supplements, and 2) the Nano-Meth group received 10 ml/l of 5% Nano-Meth starting from 1 day old until 35 days old (the end of the experiment). Nano-Meth improved final body weight, weight gains, feed intake, and feed conversion ratio. Compared to the control group, Nano-Meth significantly lowered the serum levels of triglyceride, cholesterol, very low-density lipoprotein, and low-density lipoprotein in chickens. Nano-Meth significantly increased the serum levels of total protein, albumin, high-density lipoprotein, and glucose more than the control group. Nano-Meth lowered the mRNA gene expression of ACC, FAS, MSTN, and CCK but increased that of GHR and IGFR-1. We concluded that supplementation with Nano-Meth enhances growth performance and decreases lipid accumulation in broiler chickens.

Short-wavelength light induces broiler’s behavioral and physiological syndrome through a misaligned eating rhythm

Previous work shows that long-wavelength light has a robust circadian rhythmic pattern in the expression of clock genes of chickens, whereas short-wavelength light leads to an arrhythmic oscillation of some clock genes (e.g., cClock, cCry1, cCry2, cPer2, and cPer3). However, knowledge about the consequences of LED lights on the physiological and behavioral phenotype was still not clear. This experiment hypothesizes that short-wavelength light disturbs chickens’ eating rhythm and leads to the wrong time to eat, resulting in metabolic syndrome. “Meihuang” broilers were housed in monochromatic LED blue light, green light, yellow light, red light, or white light with a very low dose (15 lx). Multiply physiological parameters were measured and the 24-h eating behavior was determined. The effects of LED light on physiological status and behavioral phenotype showed a wavelength-dependent manner. Short-wavelength light significantly decreased the level of total triglycerides and total cholesterol but increased triiodothyronine concentration. Inversely, long-wavelength light increased the triglycerides and total cholesterol and reduced the level of triiodothyronine. Further, it was found that short-wavelength light significantly boosted body weight compared with long-wavelength light, despite equivalent levels of food intake. Short-wavelength light-induced 23.4% and 14.1% of food consumption during subjective nights, but long-wavelength light did not. These results imply that when chickens eat matters, not just what they eat. Thus, low as 15 lx of blue light exposure during the typical dark period is sufficient to lead an individual to eat at the “wrong” time, causing metabolic dysfunction. Blue light should be cautiously considered to be used in the poultry breeding process. Keywords: short-wavelength light, broiler, behavior, physiology, environmental control, circadian rhythm, intermittent eating DOI: 10.25165/j.ijabe.20221503.6456 Citation: Yang Y F, Liu Q, Wang S Y, Zeng L, Pan C H, Jado A, et al. Short-wavelength light induces broiler’s behavioral and physiological syndrome through a misaligned eating rhythm. Int J Agric & Biol Eng, 2022; 15(3): 47–54.

Effects of probiotic feed additives (biosol and Zemos) on growth and related genes in broiler chickens

The goal of this study was to look into the effect of two probiotics as feed additives: Biosol (lipotropic factor containing probiotics) and/or Zemos, and to determine the best regimen for using both probiotics to improve growth performance, carcass traits, blood parameters, and transcription of some growth and immunity-related genes in broiler chickens. A commercial hatchery provided 400 one-day-old Cobb broiler chicks. The chicks were divided into four equal groups at random, each with 100 birds. Group 1 served as a negative control. Group 2 was given Probiotic 1(Biosol) in drinking water at a dose of 120 g/10,000 chick each day. Group 3 received probiotic 2 (Zemos) at a dose of 0.25 mL/L of drinking water for 3 days/week. Group 4 was given combined probiotics (Biosol + Zemos), with the same weekly dose of each probiotic alternatively. During the experiment, all chickens were fed and hydrated ad libitum and exposed to 24 h of light. Our findings revealed that chicks fed on diets supplemented with Biosol and Biosol + Zemos showed an increase in body weight gain and lower feed conversion rates. Total protein levels were elevated while cholesterol and triglyceride levels dropped, whereas, no effect was detected in the uric acid levels among those groups. Furthermore, the carcass traits of Biosol and Biosol + Zemos increased as a result of the dietary supplementations, however, the weight of the drumstick showed no difference among the four groups. Both probiotics influenced the cecal count of lactobacilli, total aerobes, E. coli, and Enterococci in, as well as the transcript levels of the MTOR, SMYD, TLR-4, and NBN genes. It is possible to conclude that supplementing broilers with probiotics and Biosol, in particular, can increase their growth performance, and improve the biochemical characteristics of the blood and transcript levels of the genes under investigation. Highlights Supplementation of the broiler diet with probiotics is essential Biosol and Zemos increased the growth performance Both probiotics improved the biochemical blood parameters Biosol up-regulated growth and immunity related genes

GHR/HindIII Locus Polymorphisms in Intron-2 GHR Gene of Papua Local Chicken

This study aimed to detect single nucleotide polymorphisms (SNPs) in intron-2 on growth hormone receptor (GHR) gene in Papua local chickens using the PCR-RFLP method to study its relationship with growth characteristics. Data on the bodyweight of 49 chickens aged 1, 2, 3, and 4 months (22 males, 27 females) and DNA samples were used for this study. The DNA fragment of size 718 bp in intron-2 of the GHR gene from the study chicken was successfully amplified using a pair of specific primers. The PCR-RFLP/HindIII analysis results found this locus's two genotypes (HindIII++ and HindIII--). HindIII+ and HindIII- alleles were 0.02 and 0.98, respectively.

Dual Host and Pathogen RNA-Seq Analysis Unravels Chicken Genes Potentially Involved in Resistance to Highly Pathogenic Avian Influenza Virus Infection.

Highly pathogenic avian influenza viruses (HPAIVs) cause severe systemic disease and high mortality rates in chickens, leading to a huge economic impact in the poultry sector. However, some chickens are resistant to the disease. This study aimed at evaluating the mechanisms behind HPAIV disease resistance. Chickens of different breeds were challenged with H7N1 HPAIV or clade 2.3.4.4b H5N8 HPAIV, euthanized at 3 days post-inoculation (dpi), and classified as resistant or susceptible depending on the following criteria: chickens that presented i) clinical signs, ii) histopathological lesions, and iii) presence of HPAIV antigen in tissues were classified as susceptible, while chickens lacking all these criteria were classified as resistant. Once classified, we performed RNA-Seq from lung and spleen samples in order to compare the transcriptomic signatures between resistant and susceptible chickens. We identified minor transcriptomic changes in resistant chickens in contrast with huge alterations observed in susceptible chickens. Interestingly, six differentially expressed genes were downregulated in resistant birds and upregulated in susceptible birds. Some of these genes belong to the NF-kappa B and/or mitogen-activated protein kinase signaling pathways. Among these six genes, the serine protease-encoding gene PLAU was of particular interest, being the most significantly downregulated gene in resistant chickens. Expression levels of this protease were further validated by RT-qPCR in a larger number of experimentally infected chickens. Furthermore, HPAIV quasi-species populations were constructed using 3 dpi oral swabs. No substantial changes were found in the viral segments that interact with the innate immune response and with the host cell receptors, reinforcing the role of the immune system of the host in the clinical outcome. Altogether, our results suggest that an early inactivation of important host genes could prevent an exaggerated immune response and/or viral replication, conferring resistance to HPAIV in chickens.

Clostridium perfringens-Induced Host-Pathogen Transcriptional Changes in the Small Intestine of Broiler Chickens.

Clostridium perfringens is an important opportunistic pathogen that may result in toxin-mediated diseases involving food poisoning/tissue gangrene in humans and various enterotoxaemia in animal species. It is a main etiological agent for necrotic enteritis (NE), the most financially devastating bacterial disease in broiler chickens, especially if raised under antibiotic-free conditions. Importantly, NE is responsible for losses of six billion US dollars annually in the global poultry industry. To investigate the molecular mechanisms of C. perfringens-induced pathogenesis in the gut and its microbiome mRNA levels in C. perfringens-infected and non-infected hosts, we used RNA sequencing technology to perform transcriptional analysis of both host intestine and microbiome using our NE model. The growth rate was significantly impaired in chickens infected by C. perfringens. In total, 13,473 annotated chicken genes were differentially expressed between these two groups, with ninety-six genes showing statistical significance (|absolute fold changes| > 2.0, adjusted p value < 0.05). Genes involved in energy production, MHC Class I antigen, translation, ribosomal structures, and amino acid, nucleotide and carbohydrate metabolism from infected gut tissues were significantly down-regulated. The upregulated genes were mainly engaged in innate and adaptive immunity, cellular processes, genetic information processing, and organismal systems. Additionally, the transcriptional levels of four crucial foodborne pathogens were significantly elevated in a synergic relationship with pathogenic C. perfringens infection. This study presents the profiling data that would likely be a relevant reference for NE pathogenesis and may provide new insights into the mechanism of host-pathogen interaction in C. perfringens-induced NE infection in broiler chickens.

Identification of novel microRNAs in Rous sarcoma Virus (RSV) and their target sites in tumor suppressor genes of chicken.

A small non-coding, evolutionarily conserved regulatory RNA molecule known as microRNA (miRNA) regulates various cellular activities and pathways. MicroRNAs remain evolutionarily conserved in different species of same taxa. They are present in all organisms including viruses. Viral miRNAs are small, less conserved and less stable and have higher negative minimal folding free energy than miRNAs of different organisms. The size of viral precursor miRNA is approximately 60-119 nucleotides in length. The structure of the mature miRNA sequences is predicted by using higher negative MFE (ΔG) value. Rous sarcoma Virus (RSV), named after its inventor Peyton Rous, has been known for causing tumors in the chicken for which it is known as an oncogenic retrovirus. Using specific criteria we have predicted 5 potential miRNAs in RSV which targeted 8 tumor suppressor genes in Gallus gallus. This study aims to predict the potential miRNAs, secondary structures and their targets for better understanding of the regulatory network of Rous sarcoma virus miRNA in forming sarcoma.

Dietary phosphorus level regulates appetite through modulation of gut and hypothalamic expression of anorexigenic genes in broiler chickens

Two experiments were designed to elucidate gut and hypothalamic molecular regulation of appetite by dietary phosphorus (P) concentration in broiler chickens. Birds (192 Cobb-500 broiler chickens) were randomly assigned to 3 experimental diets in experiment 1 (Exp. 1) and 24 broiler chickens were randomly assigned to 3 treatment groups in Exp. 2. Each diet comprised 8 replicate cages, with either 8 birds (Exp. 1) or 1 bird (Exp. 2) per replicate cage. In Exp. 1, diets contained 1.2 (P-deficient), 2.8 (P-marginal) or 4.4 (P-adequate) g/kg non-phytate P (nPP). In Exp. 2, birds fed the P-adequate diet were pair-fed (PF) to the feed consumption levels of birds fed the P-deficient diet. Feed intake and BW gain (P < 0.001) decreased in birds fed the P-deficient diet in Exp. 1. Birds fed the P-deficient diet had similar feed intake and BW gain with PF group fed the P-adequate diet (Exp. 2) but was significantly lower (P < 0.001) than birds fed the P-adequate diets. Sodium-phosphate cotransporter (NaPi-IIb) mRNA was upregulated (P < 0.05) in both experiments. Conversely, cholecystokinin (CCK) mRNA was downregulated (P < 0.01) in birds fed P-deficient diets. Anorexia-related hypothalamic cholecystokinin receptor (CCKAR) and melanocortin receptors (MC3R and MC4R) were upregulated (P < 0.05) in birds fed P-deficient diets, in both experiments. The current data show that dietary P deficiency decreases feed intake in broiler chickens by altering the expression of anorexigenic genes in the gut and hypothalamus of broiler chickens.

Virulence Comparison of Salmonella enterica Subsp. enterica Isolates from Chicken and Whole Genome Analysis of the High Virulent Strain S. Enteritidis 211.

Background: Salmonella enterica is one of the common pathogens in both humans and animals that causes salmonellosis and threatens public health all over the world. Methods and Results: Here we determined the virulence phenotypes of nine Salmonella enterica subsp. enterica (S. enterica) isolates in vitro and in vivo, including pathogenicity to chicken, cell infection, biofilm formation and virulence gene expressions. S. Enteritidis 211 (SE211) was highly pathogenic with notable virulence features among the nine isolates. The combination of multiple virulence genes contributed to the conferring of the high virulence in SE211. Importantly, many mobile genetic elements (MGEs) were found in the genome sequence of SE211, including a virulence plasmid, genomic islands, and prophage regions. The MGEs and CRISPR-Cas system might function synergistically for gene transfer and immune defense. In addition, the neighbor joining tree and the minimum spanning tree were constructed in this study. Conclusions: This study provided both the virulence phenotypes and genomic features, which might contribute to the understanding of bacterial virulence mechanisms in Salmonella enterica subsp. enterica. The first completed genomic sequence for the high virulent S. Enteritidis isolate SE211 and the comparative genomics and phylogenetic analyses provided a preliminary understanding of S. enterica genetics and laid the foundation for further study.

Genetic Polymorphisms of Chicken Antiviral Mx1 and TVB Genes in Indigenous and Giriraja Chicken

Background: Recent developments in molecular genetics lead to addressing certain poultry diseases via breeding for disease resistance. The present study was carried to identify and compare the genetic polymorphism in Chicken Mx1 and TVB genes among the indigenous and Giriraja chicken using PCR-RFLP technique. Methods: Blood samples were collected from 50 indigenous and 50 Giriraja birds and DNA isolation was done by Phenol: Chloroform: Isoamyl alcohol method. PCR amplification of Chicken Mx1 (exon 14) and Chicken TVB (exon 3) genes was carried out followed by RFLP analysis. Result: PCR product sizes of 301 bp and 303 bp of Mx1 and TVB genes, respectively were successfully amplified. RFLP analysis of Mx1 gene with Hyp8I restriction enzyme revealed three genotypes AA, AB and BB. In indigenous birds genotypic frequencies of AA, AB and BB were 0.314, 0.493 and 0.194, respectively and gene frequencies were 0.56 and 0.44 for alleles A and B, respectively. In Giriraja birds, genotypic frequencies for AA, AB and BB were 0.27, 0.499 and 0.23, respectively and gene frequencies were 0.52 and 0.48 for alleles A and B, respectively. RFLP analysis of TVB gene with NlaIII restriction enzyme revealed two genotypes viz., AA and AB. In indigenous birds genotypic frequencies of AA and AB were 0.81 and 0.18, respectively and gene frequencies were 0.9 and 0.1 for alleles A and B, respectively. In Giriraja birds genotypic frequencies for AA and AB were 0.774 and 0.211, respectively and gene frequencies were 0.88 and 0.12 for alleles A and B, respectively.

Immune-Related Gene Expression Profiling of Broiler Chickens Fed Diets Supplemented with Vinification Byproducts: A Valorization Approach II.

Simple SummaryThe valorization of grape byproducts appears to be a sustainable bioeconomic strategy that could promote the substitution of synthetic with natural antioxidant compounds in the food and feed industry. The nutritional physiology of broiler chickens requires special attention to the interactions between nutrients and antioxidant mechanisms since the stressor signals of factory farming could impair the immune status, resulting in detrimental effects on broilers’ performance. The objective of this study was to assess the inclusion of grape byproducts (grape pomace, wine lees, and stem extract) on the transcriptional profiling of genes regulating the immune system in the liver, bursa of Fabricius, and spleen. The dietary supplementation of grape byproducts rich in polyphenolic compounds decreased the mRNA levels of the predominant pro-inflammatory receptor in the liver, while in the spleen, the stem extract diet upregulated the aforementioned receptor. Upregulation of interleukin 8 was observed in the bursa of Fabricius and spleen of the stems extract-fed broilers. Although grape byproducts depicting a sustainable source of bioactive compounds with vast antioxidant potential, there were unveiled preliminary insights for immune stimulation at the transcriptional level.The valorization of vinification byproducts portrays a promising bioprocess for the enrichment of animals’ diet with bioactive compounds, such as polyphenols, which could regulate the immune response. Therefore, the impact of dietary grounded grape pomace (GGP), wine lees extract (WYC), and grape stem extract (PE) on the relative transcript level of immune related genes of broiler chickens were examined. Two hundred forty, one-day-old as hatched (male/female) chicks (Ross 308) were allocated to four dietary groups, with four replicate pens each with 15 birds. Birds were fed either a basal diet (CON) or the basal diet supplemented with 2.5% GGP, or 0.2% WYC, or 0.1% PE for 42 d. The relative expression of immune-related genes was investigated using a real-time PCR platform. The mRNA levels of Toll-like Receptor 4 (TLR4) were downregulated (p = 0.039) in the liver of broilers fed the GGP-containing diet compared to the CON, while in the spleen of PE-fed broilers, TLR4 was significantly upregulated (p = 0.043). The mRNA levels of interleukin 8 (IL8) tended to upregulate (p = 0.099) in the bursa of Fabricius and were significantly increased (p = 0.036) in the spleen of broilers fed the PE diet. Vinification byproducts depict a promising sustainable source of polyphenols for the poultry feed industry, but more research is needed under field conditions.

Insights Into mRNA and Long Non-coding RNA Profiling RNA Sequencing in Uterus of Chickens With Pink and Blue Eggshell Colors

The blue egg is both of biological interest and economic importance for consumers, egg retailers, and scientists. To date, the genetic mechanisms underlying pigment have mainly focused on protein-coding genes. However, the underpinning mechanism of non-coding RNAs on the pigment deposition among different eggshell colors remains unknown. In this study, RNA sequencing was employed to profile the uterine gland transcriptome (lncRNA and mRNA) of 15 Changshun blue eggshell layers, to better understand the genetic mechanisms of deposition of blue eggshell color. Results showed that differentially expressed mRNAs, GO terms, and KEGG pathways among pink-eggshell and blue-eggshell chickens were mainly targeting immune- and transporter-related terms with the SLC family, IgJ, CD family, and MTMR genes. Furthermore, the progesterone-mediated oocyte maturation and cortisol synthesis and secretion pathway with targeted gene PGR and Pbx1 were significantly enriched between blue- and pink-eggshell chickens. Integrating analysis of lncRNA and mRNA profiles predicted 4 and 25 lncRNA–gene pairs by antisense and cis analysis. They were relative to immune, nerve, and lipids and amino acid metabolisms, porphyrin, and chlorophyll metabolism with targeted gene FECH and oxidative phosphorylation and cardiac muscle contraction pathways with targeted gene COX6A1. Within blue-eggshell chickens, the GO terms hindbrain tangential cell migration and phosphatidylinositol monophosphate phosphatase activity with targeted gene Plxna2 and MTRM1 were identified. Integrating analysis of lncRNA and mRNA profiles predicted 8 and 22 lncRNA–gene pairs. Most pathways were mainly enriched on lipid-related metabolisms as found in mRNA sequencing. The lncRNAs did exert similar functions in color formation by modulating pigment disposition and immune- and lipid-related metabolisms. Our results provide a catalog of chicken uterine lncRNAs and genes worthy of further studies to understand their roles in the selection for blue eggshell color layers.