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Toll-like receptors (TLRs) are one of the important immunes signaling pathways that participate in the activation of host immune response after detecting microbial pattern molecules. Despite important expression profile TLRs genes in chicken erythrocytes, studies have been lacking. This study investigated the expression profile of TLRs pathway immune gene in chicken erythrocytes in response to Mycoplasma synoviae. The purpose of the current in-vitro study was to determine the chicken erythrocytes interaction with M. synoviae using a transmission electron microscope (TEM). The mRNA gene expression of TLR1, 2, 3, 4, 5, 7, 15, MHC I, II, and MyD88 in M. synoviae infected chicken erythrocytes was determined using quantitative real-time PCR (qRT-PCR) at four different time intervals (0, 2, 6 and 10 h) post-infection and compared to uninfected controls. The mRNA expression of TLRs such as TLR1, 2, 3, 15, and MHC II were significantly upregulated at 6 and 10 h post-infection in infected chicken erythrocytes. However, significantly upregulated expression of TLR5 and MHC I were noted at 2, 6, and 10 h while TLR4 and MyD88 mRNA expression was also significantly upregulated but at different time intervals. This study provides the first evidence of upregulated expression of TLR signaling pathway genes in M. synoviae infected chicken erythrocytes. These results provide new insights on M. synoviae infection resistance mechanisms and the role of TLR signaling immune genes in the control of the host immune response. [ABSTRACT FROM AUTHOR]

Tibial dyschondroplasia (TD) is one of the most common problems in broiler chickens. Chicken erythrocyte is the most abundant cell found in blood vessels, in which host defense resistance (HDR) and pattern recognition receptors (PRRs) play a significant role in the immune response. However, the function of HDR and PRRs in thiram induce TD chicken is not fully known yet. Therefore, this study was designed to evaluate the effect of recombinant glutathione-S-transferase A3 (rGSTA3) protein on HDR and PRRs in the thiram-induced TD chicken. One hundred twenty arbor acres (AA+) broiler chickens of seven-day-old were equally divided into six groups. Group A, B, and C were treated with 0, 20, 50 μg.kg-1 of rGSTA3 protein, respectively. Group D, E and F were treated with 0, 20, 50 μg.kg-1 of rGSTA3 protein + thiram 100 μg.kg-1. The results showed that chicks treated with thiram showed a higher TD score with typical lesions in the proximal growth plate than the rGSTA3 supplementary group. A morphological characteristic of TD was measured by TD index. Chicken erythrocytes constitutively expressed transcripts for HDR and PRRs genes. Moreover, the quantitative real time PCR (qRT-PCR) results showed that HDRs and PPRs expressions were suppressed by thiram, which found upregulated in rGSTA3 protein supplementary group. These findings demonstrated that mRNA expressions of HDRs and PPRs are highly related to thiram. Moreover, rGSTA3 protein increased the immune response of HDRs and PPRs. Conclusively the results of this study provide the strong evidence that rGSTA3 protein can promote an immune response in chicken erythrocytes by up regulating the expression of HDR and PRRs genes. [ABSTRACT FROM AUTHOR]

Mycoplasma synoviae is one of the most important pathogens in the poultry industry and often causes diseases of a chronic and persistent nature. However, there is limited data available on Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway genes expression in chicken erythrocytes infected with Mycoplasma synoviae (M. synoviae). Therefore, the aim of the current in-vitro study was to determine the interaction between chicken erythrocytes and M. synoviae using Transmission electron microscope (TEM) and further to investigate the mRNA gene expression of MyD88 (Myeloid differentiation primary response 88), CCL5 (C-C Motif Chemokine Ligand 5), MDA5 (melanoma differentiation-associated protein 5) IKBKE (inhibitor of nuclear factor kappa-B kinase subunit epsilon), NFKBIA (NF-kappa-B inhibitor alpha), NFKBIE (NF-kappa-B inhibitor epsilon), Interferon Alpha (IFN-α), cMGF (chicken myelomonocytic growth factor), and TRAF6 (Tumor necrosis factor receptorassociated factor 6) in chicken erythrocytes infected with M. synoviae using quantitative real-time PCR (qRT-PCR) at four different time intervals (0, 2, 6 and 10 h) post-infection and compared to uninfected controls. The results indicated that M.synoviae interacted efficiently in chicken erythrocytes, which strongly induced the up-regulation of NF-κB pathway and other immune system genes in response to early bacterial infection such as NFKBIA, NFKBIE, IKBKE, CCL5, MDA5, MyD88, and TRAF6 at different h's interval. Whereas cMGF and IFN-α expression were significantly downregulated during early time intervals such as 0 h, 2 h and 6 h while later on the expression level significantly increased. These results will lead to increased insights on M. synoviae infection resistance mechanisms and the role of NF-κB signaling pathway and other immune system genes in the control of the host immune response. [ABSTRACT FROM AUTHOR]

Carcass weight, as a measure of meat yield, and body measurements are directly correlated traits in livestock. However, longitudinally collected phenotype records of local breeds are not comprehensive. The research was performed on Qinchuan bull population to understand their growth and development, and data from Qinchuan bull that was weighed and measured at birth, 6, 12, 18, and 24 months of age was analyzed. Furthermore, Logistic, Brody, Gompertz, and Bertallanffy were used to fit the growth curves for weight and body size traits. The results showed that the four curve models have good fitting degrees for the weight and body size (R2 > 0.99), and the Bertallanffy model exhibited a good fit to the measured data of body weight, and the model estimated the inflection point of body weight as (5.43 months of age, 122.01 kg). Particularly, the limited mature body weight can reach 557.8 kg by the Brody model. Body weight was significantly positively correlated with body height, hip height, body length, chest circumference, abdominal girth, and calf girth (p < 0.0001), and the correlation between body weight and body length was the highest (r = 0.975). The regression equation predicting body weight was Y = −275.691 + 3.28 X3 + 1.311 X4 − 0.397 X5. [ABSTRACT FROM AUTHOR]

For its role in the mediation of myoblast proliferation, fibroblast growth factor receptor 1 (FGFR1) was considered a functional candidate gene for growth performance in Tibetan sheep. Via the polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) approach, four single nucleotide polymorphisms (SNPs) including g.14752C > T (intron 1), g.45361A > G (intron 7), g.49400A > G (3'UTR region) and g.49587A > T (3'UTR region), were identified in 422 ewes. The association analysis demonstrated that individuals carrying the AA genotype of g.49400A > G had significantly greater withers height, length than those with GG genotype (p < 0.05). Individuals with genotype AA of g.49587A > T had significantly greater weight and chest circumference than those with genotype TT (p < 0.01). Additionally, the individuals with Hap1/1 diplotypes (CAAA-CAAA) were highly significantly associated with weight and chest circumference than Hap1/2 diplotypes (CAAA-CAAT) (p < 0.05). The quantitative real-time polymerase chain reaction (qPCR) analysis revealed that the FGFR1 was detectable expressed in muscle tissues within three different age stage. Remarkably higher mRNA expression was detected at fetal lamb stage as compared with adult ewes (p < 0.01). The outcome of this research confirmed that both g.49400A > G and g.49587A > T of FGFR1 were involved in growth-related traits, which may be considered to be genetic markers for improving the growth traits of Tibetan sheep. [ABSTRACT FROM AUTHOR]

Aim and Background. Tibial dyschondroplasia (TD) is a tibiotarsal bone disorder of rapid growing poultry species. The complete pathogenicity of TD remains unknown; therefore, a study was conducted to screen possible role of proliferation-related gene expression in thiram-induced tibial TD chickens. Materials and Methods. Three hundred sixty (n = 360) broiler chickens were equally divided into control (C) and thiram (T) groups. Furthermore, the C and T groups were dividedinto 8-, 9-, 11-, and 13-day-old chickens. Results. Clinically, it was observed that broiler chickens of group T had abnormal posture, gait, and lameness, and histopathological results revealed dead and abnormal chondrocytes of T group on day 6. Real-time qPCR results showed that HDAC1, MTA1, H4, and PCNA genes were significantly expressed (P < 0.05). HDAC1 was upregulated on days 1, 2, 4, and 6 (P < 0.01); MTA1 was upregulated on days 1 and 2 (P < 0.01); H4 was upregulated on days 2 and 4 (P < 0.01), and PCNA was downregulated on days 1, 2, and 4 (P < 0.01). Furthermore, IHC results of HDAC1 protein were significantly (P < 0.01) expressed in proliferative zone of day 1 and hypertrophic zone of day 6. MTA1 protein was significantly (P < 0.01) expressed on days 1, 2, and 6 in all zones, except prehypertrophic zone of day 2. Conclusion. In conclusion, the mRNA expressions of HDAC1, MTA1, H4, and PCNA were differentially expressed in the chondrocytes of thiram-induced TD chickens. HDAC1 and MTA1 protein expression found involved and responsible in the abnormal chondrocytes' proliferation of broiler chicken. [ABSTRACT FROM AUTHOR]

Avian influenza virus (H9N2) is one of the major cause of illness in chickens with potential threats to human health. Chicken erythrocytes are known as most abundant circulating cells in the body which participate in inflammation and immune responses. Influenza viruses interact with the host cells at cell surface, leading to cell death in human and animals through apoptosis. Current study was designed to investigate the expression levels of apoptosis-related genes in the H9N2 infected chicken erythrocytes. qRT-PCR and Western blot techniques were performed to determine the expression levels of apoptosis-related genes including Bax, Bcl-2 BAG-1, BAG-3 and STAT3 in H9N2 infected erythrocytes of chickens for incubation time intervals of 0, 2, 6 and 10 h. The expression of all the genes related to apoptosis were significantly up-regulated at different time periods, except of Bcl-2 which was significantly down-regulated at 6 and 10 h of infection. To conclude, there are alterations in expression levels of apoptosis-related genes in chicken’s erythrocytes at different time intervals that are highly related to the development of H9N2 infection. This study may provide a new biomarker to detect H9N2 infection in chickens. [ABSTRACT FROM AUTHOR]

Marek's disease virus (MDV) is a highly cell-associated, oncogenic alpha herpesvirus that causes Marek's disease (MD) in chickens. Chicken erythrocytes are nucleated cells of myeloid origin which contribute to gases exchange, transportation and immunity. The objective of this study was to reveal the immune responses of MDV-infected erythrocytes of chicken by transcriptome analysis. Twenty specific pathogenfree (SPF) chickens were divided into two groups as control and treatment. The chickens of treatment group were inoculated with MDV, and blood samples of two groups were obtained at 14 and 22 days post-infection (dpi) for RNA extraction. Transcriptome analysis was performed to explore the immune responses of chicken erythrocytes to MDV infection at 14 dpi and 22 dpi. In total, 129.6 million reads of two groups at two time points were mapped against the reference genome (Gallus gallus 5.0). A significant differential pattern of the immune response of chicken erythrocytes was identified by the GO and KEGG databases between the two phases of MDV infection. The results indicated that cytokine-cytokine receptor interaction, toll-like receptor signaling pathway, mitogen-activated protein kinases (MAPK) signaling pathway, Janus kinase/signal transducers and activators of transcription (JAK-STAT) signaling pathway were involved in the response to MDV infection. The expression levels of all most differentially expressed genes (DEGs) in these four pathways were upregulated significantly at 14 and 22 dpi, indicating that the immune responses of erythrocytes were induced by MDV. This study may help to elucidate the molecular mechanism of chicken erythrocytes relate to immune responses in MDV infection. [ABSTRACT FROM AUTHOR]

This paper presented the results on the study of insulin receptor substrate 1 (IRS1) polymorphisms in Chinese black Tibetan sheep. Via DNA direct sequencing, four variations within 3′ untranslated region (UTR) of IRS1, including g.9382T > G, g.9569T > G, g.9572C > T and g.9695A > C were detected in the black Tibetan sheep population. Based on the χ2 test, those four loci deviated from Hardy–Weinberg equilibrium (p < 0.05). In g.9569T > G locus, genotype of GG possessed advantage on body weight (p < 0.05). In g.9572C > T locus, individuals with genotype of TT homozygous mutation decreased significantly on body weight, withers height, body length and chest circumference (p < 0.05 or p < 0.01). In g.9695A > C locus, the body weight and chest circumference were also higher in AA carriers than in CC carriers (p < 0.05). Our results provided evidence that polymorphisms in IRS1 were associated with growth efficiency traits by quantitative genetic analysis, and may be used for marker-assisted selection in Chinese indigenous sheep. [ABSTRACT FROM AUTHOR]

Chicken's erythrocytes are most abundant cells circulating in the body which participate in inflammation and immune responses. However, expression levels of prostaglandins-related genes in H9N2 infected chicken's erythrocytes have not been studied yet. So, the present study was designed to investigate the PG-related genes in erythrocytes of H9N2 infected chickens. In current study, qRT-PCR and Western blot analysis were performed to identify the expression levels of PG-related genes (COX-2, GSTA3, PTGDS, PTGES, PTGER3 and PTGER4) in erythrocytes of H9N2 infected chickens at time intervals of 0, 2, 6 and 10 h and P < 0.05 was considered as statistically significant. Expression levels were increased significantly in case of COX-2 at 0 and 6 h, GSTA3 and PTGDS at 0 h and PTGER4 at 2 h of infection, while decreased expression was observed in case of COX-2 at 2 h, GSTA3 at 10 h, PTGDS at 6 and 10 h, PTGES at 2 and 10 h, PTGER3 at 0, 2, 6 and 10 h and PTGER4 at 6 and 10 h of infection. Besides, western blot analysis of COX-2 protein expression further supported the mRNA results. Moreover, this was first study indicating the expression levels of PG-related genes in the H9N2 infected chicken's erythrocytes. This study may provide a new biomarker to detect H9N2 in chickens. Future studies are in process to know the morphological features, and to evaluate the mechanism exhibited by erythrocytes in response to H9N2 with new experimental evidence in chickens. [ABSTRACT FROM AUTHOR]

Chicken erythrocyte, which is the most abundant cell subset in blood vessel, also exists in cartilage zone. It has recently been shown that chicken erythrocytes constitutively express toll-like receptors (TLRs), cytokines, and avian β-defensins (AvBDs). Host defense peptides (HDPs) are important effector molecules in innate immunity. In this study, thiram has been used to induce tibial dyschondroplasia (TD) and the chickens were injected with different doses of recombinant glutathione-S-transferase A3 (rGSTA3) proteins. We explored the responses of HDPs in erythrocytes to thiram-induced TD and rGSTA3 protein by quantitative real time PCR (qRT-PCR). The results showed many HDPs expressions were suppressed by thiram-induced TD, and the expressions of HDPs were upregulated by rGSTA3 protein. These findings demonstrated that mRNA expressions of HDPs are highly related to thiram-induced TD, and rGSTA3 protein enhanced the immune response of chicken erythrocytes by up-regulating HDPs expression. Future studies may be aimed at elucidating the association between the expression of these peptides and the development of the TD. [ABSTRACT FROM AUTHOR]

The research was conducted to investigate the immunomodulatory and growth promoting effects of basil (Ocimum basilicum) and ascorbic acid in heat stressed broiler chicken. A total of 360 (Hubbard) oneday old broilers were reared under heat stress (38 ± 1°C) and randomly divided into three groups i.e. A, B and C as control, basil seed at 5g/kg and ascorbic acid at 200 mg/kg supplementation, respectively. Improvement of intestinal villus seen in basil supplementary group (P>0.05), and significant increase in the absorption of crude protein (CP), crude fibre (CF), and metabolized energy (ME). However, the values of red blood cells and packed cell volume were non-significant whereas white blood cells, haemoglobin, and new castle disease antibody titer were significantly higher in basil supplementary group. The weight gain and feed conversion ratio significantly improved in basil treated group, while ascorbic acid and basil significantly decreased the water intake. Body temperature and mortality percentage were significantly decreased in ascorbic acid and basil supplementary group as compared to control. Dressing percentage significantly increased in both supplementary groups. It is concluded that supplementation of basil at 5g/kg feed, promotes the growth, improves the intestinal villus size, feed efficiency and immunity of heat stressed broiler chicken. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is one of the most toxic metals released into the environment. Here, we investigated the protective role of Zn 2+ and/or N-acetyl- L -cysteine (NAC) against Cd cytotoxicity in the erythrocytes of Arbor Acres (AA) broiler chickens. Four hundred one-day-old AA chickens were divided into 12 groups for in vitro and in vivo studies. Zn 2+ and/or NAC was given to the Cd exposed AA chickens to assess their protective roles. This was accomplished by investigating nuclear morphological abnormalities, oxidative stress (SOD, CAT, GPx, GSH and T-AOC), cell apoptosis, ROS accumulation and mitochondrial membrane potential (MMP). Results showed that Cd led to dose- and time-dependent cytotoxicity in the erythrocytes of AA chickens characterized by morphological abnormalities, nucleus damage, increased apoptosis rate and antioxidants depletion. Zn 2+ or NAC significantly decreased the erythrocyte apoptosis, ROS production and mitochondrial membrane depolarization caused by Cd. SOD, CAT, GPx, GSH and T-AOC activities significantly decreased both in serum and erythrocytes of Cd exposed AA chickens. The supplementation with Zn 2+ or NAC alleviated Cd induced oxidative stress through promoting SOD or GPx/GSH activities respectively. NAC presented a better role in reducing apoptosis, improving antioxidant activities more than Zn 2+ in vitro. The combined use of Zn 2+ and NAC enhanced cytoprotection in Cd exposed erythrocytes of AA chickens compared to Zn 2+ or NAC alone. In conclusion, Zn 2+ and NAC exerted remarkable protective roles in Cd exposed erythrocytes of AA chickens by inhibiting cell apoptosis and oxidative stress, and this provides a promising approach to antagonize Cd poisoning in poultry. [ABSTRACT FROM AUTHOR]

• TD is an avian welfare problem cause lameness in fast-growing broiler chickens. • Therapeutic role of Celecoxib investigated in thiram-induced TD chickens. • The expression level of COX-1/2, MMP-13 and ColX genes were investigated. • Celecoxib plays a therapeutic role in the welfare problem of TD. • Celecoxib supplementation prevented chondrocytes of thiram induced TD chickens from damage. The pathogenicity of tibial dyschondroplasia (TD) is unknown; therefore, it has not been completely evacuated. In this study, we planned to find the possible therapeutic role of Celecoxib supplementation in thiram-induced TD chickens. Three hundred sixty (n = 360) broiler chickens were divided into 3 groups as control (C), thiram (T) and Celecoxib supplementary group (ST) for day 1 (8 day old), 2 (9 day old), 4 (11 day old) and 6 (13 day old). Thiram 100 mg·kg−1 was given in group (T) along with basal diet to induce TD. Supplementation of Celecoxib at 40 mg·kg−1 was given in group (ST) along with the basal diet and thiram. Clinically it was observed that broiler chickens of group T had more difficulty in locomotion than group ST. Histopathological results revealed that damaged chondrocytes increased from day 1 to 6 in group T. However, Celecoxib supplementation to thiram induced TD chickens reduced the tibial lesions significantly. Cyclooxygenase-1 (COX-1) gene was highly expressed at different stages (P < 0.01) while its protein expression was seen at a lower level. Cyclooxygenase-2 (COX-2) gene and protein expression were also changed at different stages. Expression of matrix metalloproteinase-13 (MMP-13) gene were observed high on day 1 (P < 0.01), which insignificantly changed on day 2 and 4. MMP-13 protein expression found reduced parallel to its gene expression (P < 0.01), on day 2. Type X collagen (ColX) gene expression was upregulated (P < 0.01). After adding Celecoxib in the diet, both COX-1 and COX-2 gene (P < 0.01) and their protein expression expressed similar to the control group, and to some extent observed parallel to MMP-13 gene (P < 0.01) & protein expression. In conclusion, changes in the expression of COX-1/2, MMP-13 and ColX might be related to TD occurrence. The dietary supplementation of Celecoxib plays a therapeutic role to overcome the welfare problem of TD. [ABSTRACT FROM AUTHOR]

The present study was designed to evaluate the efficiency of herbs on the immunity related-genes and health of broiler chickens challenged with coccidiosis. Total (one-day-old, 450) broiler chickens were allotted into 9 groups (n = 50/group). Group A controlled without supplementation, Group B, C, D, E, F, and G were supplemented with Aloe barbadensis (5ml/L), Tinaspora Cordifolia (20g/L), Bambusaarun-dinacea (5g/L), Emblica officinalis (10g/L), Ferulafoetida regal (500mg/L), Tamarindusindica (50mg/L) respectively. Group H and I were supplemented with a mixture of all 6 herbs/plants at 2ml/L; mixed with distilled water and another with citric acid. Anticoccidial effect of different herbs on comparative average values of bloody diarrhea score, lesion score, oocysts score, and survival percentage were recorded. The survival rate of coccidiosis challenged broiler chickens were observed significantly (P<0.05) high in group B, H, and I. Higher level of intestinal histomorphology, blood hematology, E. coli, and Lactobacillus observed (P<0.05) in group H and I. Antibody titer results (ND and H7) suggested that the titer was high for group B and I. Hormone profiling results of T3, T4 and AST was observed higher in group H. mRNA expression of toll-like receptors 1,2, 3, 4, 5 (TLR1, TLR2, TLR3, TLR4, TLR5) genes were validated on RT-PCR, which were significantly different (P<0.05) upregulated in group H and I. In conclusion, supplementation of herbs mixture (Group H and I) at the dose 2ml/L to the coccidiosis challenged broiler chickens overall improved the health and immunity by regulating the mRNA expression of immunity-related toll-like receptors. [ABSTRACT FROM AUTHOR]

• mRNA expression of HSL gene was quantified different tissue in two age groups of Qinchuan cattle. • Five SNPs of bovine HSL gene were identified within exon. • Effects of single markers/haplotype combinations on intramuscular fat content were analyzed. Hormone-sensitive lipase (HSL) was considered as an essential enzyme in glucolipid metabolism. It has been proposed to be a lead candidate gene for genetic markers of lipid deposition in livestock. The aim of this study was to identify sequence variants (SVs) of the bovine HSL gene and evaluate the relations to intramuscular fat in two indigenous Chinese beef cattle breeds. Expression analysis by quantitative real-time polymerase chain reactions (qPCR) indicated that expression levels of bovine HSL gene were highest in the perirenal fat and heart within two different age stage (adult and calf), respectively. Five SVs were identified by direct DNA sequencing, which included four missense mutations (g.16563C>T, g.16734G>A, g.16896A>G, g.17388G>T) in exon 8 and a synonymous mutation (g.17402C>T) in exon 9. Population genetic analysis showed that except for g.16563C>T and g.17402C>T, all the other detected SVs strongly affected the bovine intramuscular fat content (P < 0.01 or P < 0.05). The individuals with Hap5/5 diplotypes (CC-GG-GG-GG-CC) was highly significantly associated with intramuscular fat content than the other diplotypes (P < 0.01). The above results suggested that the HSL gene can used as potential candidate markers gene for the beef breed improvement through marker assisted selection in Chinese cattle breeds. [ABSTRACT FROM AUTHOR]

The therapeutics having ability to target cancer cells specifically and exhibit nominal cytopathic effect on normal healthy cells are highly significant for cancer therapeutic applications. Recombinant porcine natural killer lysin (rpNK-lysin) has proven cationic anti-bacterial and anti-tumor peptide. Herein, we report its anti-invasion and anti-metastasis effects on hepatocellular carcinoma (HCC) cells in vitro. We first investigate the maximum non-toxic concentration (MNTC) of rpNK-lysin for the normal hepato cells (L-02). Using MNTC rpNK-lysin, we explore anti-proliferative, anti-adhesive, anti-invasive and anti-metastatic effect of rpNK-lysin on three different HCC cells lines (SMMC-7721, 97-H and HepG2) through MTT, wound-healing, adhesion and invasion assay along with mRNA and protein expression. The results reveal that rpNK-lysin has potential to specifically inhibit HCC cells growth in a dose and time-dependent manner with a little cytopathic effect on the L-02 cells, effectively reduce migration, adhesion and invasion ability of HCC cells. rpNK-lysin significantly reduce Fascin1 expression, which subsequently decrease β-catenin expression and metaloproteinases (MMP-2 and MMP9). This study suggest that MNTC rpNK-lysin has an anti-invasion and anti-metastasis effect on HCC cells in vitro through inhibition of Fascin 1 expression which regulates Wnt/β-catenin signaling pathway by inducing β-catenin degradation and subsequently results in suppression of MMP-2 and MMP9 expression. [ABSTRACT FROM AUTHOR]