RESUMEN Introducción: El extracto de cereza tiene una gran cantidad de antocianinas y flavonoides con efectos antioxidantes. Se ha demostrado que sus altas características antioxidantes reducen los marcadores de dolor muscular retardado (DOMS) y el daño muscular inducido por el ejercicio (EIMD) para mejorar la recuperación después del ejercicio. Objetivo: Verificar los efectos del extracto de cereza en el daño muscular posterior al ejercicio. Métodos: Se realizaron búsquedas sistemáticas en Google scholar, Medline y Scopus hasta febrero de 2022. Se aplicó la herramienta de colaboración Cochrane para determinar los riesgos de sesgo. Resultados: Los resultados mostraron que la administración de extracto de cereza no tuvo un impacto decreciente en los niveles de creatina quinasa en general: (WMD = 12,85 UI. L-1, IC del 95%: −35,94, 61,64; P = 0,606). Se observó una considerable heterogeneidad entre los artículos (prueba Q de Cochran = 990,80, P = 0,000, I2 = 96,7 %). Sin embargo, el consumo de extracto de cereza tiene un efecto significativo de reducción del dolor (WMD = −6,105 mm; IC del 95%: −11,193 −1,017; p = 0,019). Conclusión: El consumo de extracto de cereza fue eficaz para reducir el dolor muscular de aparición tardía entre los participantes en el análisis global y de subgrupos. Así, el extracto de cereza puede ser una alternativa complementaria en la recuperación después de los ejercicios. Nivel de evidencia II; Estudios terapéuticos - Revisión de manuscritos. Introducción DOMS (DOMS EIMD (EIMD Objetivo Métodos scholar 2022 sesgo Resultados general WMD 1285 12 85 12,8 UI L1, L1 L 1, 1 L-1 95% 95 3594 35 94 −35,94 61,64 6164 61 64 0,606. 0606 0,606 . 0 606 0,606) prueba 99080 990 80 990,80 0000 000 0,000 I 967 96 7 96, %. % %) embargo 6105 6 105 −6,10 mm 11193 11 193 −11,19 −1,017 1017 017 0,019. 0019 0,019 019 0,019) Conclusión subgrupos Así ejercicios II manuscritos 202 128 8 12, L- 9 359 3 −35,9 61,6 616 060 0,60 60 9908 99 990,8 00 0,00 610 10 −6,1 1119 19 −11,1 −1,01 101 01 001 0,01 20 −35, 61, 06 0,6 990, 0,0 −6, 111 −11, −1,0 2 −35 0, −6 −11 −1, −3 − −1

RESUMO Introdução: O extrato de cereja tem uma alta quantidade de antocianinas e flavonóides contendo efeitos antioxidantes. Suas altas características antioxidantes demonstraram reduzir os marcadores de dor muscular retardada (DOMS) e dano muscular induzido pelo exercício (EIMD) para melhorar a recuperação após o exercício. Objetivo: Verificar os efeitos do extrato de cereja nos danos musculares pós-exercício. Métodos: Google scholar, Medline e Scopus foram sistematicamente pesquisados até fevereiro de 2022. A ferramenta de colaboração da Cochrane foi aplicada para determinar os riscos de viés. Resultados: Os resultados mostraram que a administração do extrato de cereja não teve um impacto decrescente nos níveis de creatina quinase em geral: (WMD = 12,85 IU. L-1, 95% CI: −35,94, 61,64; P = 0,606). Uma heterogeneidade considerável foi observada entre os artigos (teste Q da Cochran = 990,80, P = 0,000, I2 = 96,7 %). Porém, há um efeito redutor significativo na sensação de dor pelo consumo de extrato de cereja (WMD = −6,105 mm; 95% CI: −11,193 −1,017; p = 0,019). Conclusão: O consumo de extrato de cereja foi efetivo na redução de dores musculares de início tardio entre os participantes, na análise geral e nos subgrupos. Assim, o extrato de cereja pode ser uma alternativa complementar na recuperação após os exercícios. Nível de evidência II; Estudos terapêuticos - Revisão de manuscritos. Introdução DOMS (DOMS EIMD (EIMD Objetivo pósexercício. pósexercício pós pós-exercício Métodos scholar 2022 viés Resultados WMD 1285 12 85 12,8 IU L1, L1 L 1, 1 L-1 95 CI 3594 35 94 −35,94 61,64 6164 61 64 0,606. 0606 0,606 . 0 606 0,606) teste 99080 990 80 990,80 0000 000 0,000 I 967 96 7 96, %. % %) Porém 6105 6 105 −6,10 mm 11193 11 193 −11,19 −1,017 1017 017 0,019. 0019 0,019 019 0,019) Conclusão participantes subgrupos Assim exercícios II manuscritos 202 128 8 12, L- 9 359 3 −35,9 61,6 616 060 0,60 60 9908 99 990,8 00 0,00 610 10 −6,1 1119 19 −11,1 −1,01 101 01 001 0,01 20 −35, 61, 06 0,6 990, 0,0 −6, 111 −11, −1,0 2 −35 0, −6 −11 −1, −3 − −1

ABSTRACT Introduction: Cherry extract has a high amount of anthocyanins and flavonoids containing antioxidant effects. Its high antioxidant characteristics have been shown to reduce markers of delayed muscle soreness (DOMS) and exercise-induced muscle damage (EIMD) to improve recovery after exercise. Objective: Verify the effects of the cherry extract on post-exercise muscle damage. Methods: Google scholar, Medline, and Scopus were systematically searched until February 2022. The Cochrane Collaboration tool was applied to determine the risks of bias. Results: The results showed that cherry extract administration did not have a decreasing impact on creatine kinase levels overall: (WMD = 12.85 IU. L-1, 95% CI: −35.94, 61.64; P = 0.606). Considerable heterogeneity was observed among the articles (Cochran's Q-test = 990.80, P = 0.000, I2 = 96.7 %). However, there is a significant reducing effect on pain sensation by the consumption of cherry extract (WMD = −6.105 mm; 95% CI: −11.193 −1.017; p = 0.019). Conclusion: Cherry extract consumption effectively reduced late-onset muscle pain among participants in the overall and subgroup analysis. Thus, the cherry extract may be a complementary alternative in recovery after exercise. Level of evidence II; Therapeutic studies - Manuscript review. Introduction DOMS (DOMS exerciseinduced exercise induced EIMD (EIMD Objective postexercise post Methods scholar Medline 2022 bias Results WMD 1285 12 85 12.8 IU L1, L1 L 1, 1 L-1 95 CI 3594 35 94 −35.94 61.64 6164 61 64 0.606. 0606 0.606 . 0 606 0.606) Cochrans Cochran s Qtest Q test 99080 990 80 990.80 0000 000 0.000 I 967 96 7 96. %. % %) However 6105 6 105 −6.10 mm 11193 11 193 −11.19 −1.017 1017 017 0.019. 0019 0.019 019 0.019) Conclusion lateonset late onset analysis Thus II review 202 128 8 12. L- 9 359 3 −35.9 61.6 616 060 0.60 60 9908 99 990.8 00 0.00 610 10 −6.1 1119 19 −11.1 −1.01 101 01 001 0.01 20 −35. 61. 06 0.6 990. 0.0 −6. 111 −11. −1.0 2 −35 0. −6 −11 −1. −3 − −1

ABSTRACT Gut microbiota plays an important role in animal health, production and diseases. Little is known about whether heat stress alters the composition of quail gut microbiota; therefore, we studied the effects of heat stress on growth performance, antioxidant functions, heat shock proteins and caecal microbiota. Two groups of 40 (20-day-old) quails were set up, including a control reared at 24 ± 2°C and a heat stress group subjected to heat stress at 36 ± 2°C for 4 h per day for 7 consecutive days. We found that heat stress significantly elevated the relative expression levels of HSP70 and HSP90 mRNA in the thymus, bursa and spleen by real-time fluorescence quantitative PCR assay, indicating successful establishment of the experimental model. Heat stress was found to have an effect on gut microbiota composition. At the genus level, Alistipes were significantly increased in the heat stress group. PICRUSt2 function prediction revealed that most of the KEGG pathways with high temperature-induced abundance differences are metabolism-related. These data indicated that heat stress reduced the production performance of quails by affecting antioxidant functions, as well as the composition and structure of the intestinal microbiota. The results of this study provide technical information for conducting research on heat stress prevention and control techniques in quails. health diseases therefore functions 20dayold dayold 20 old (20-day-old up 2 2C C 3 days HSP HSP7 HSP9 thymus realtime real time assay model level PICRUSt temperatureinduced temperature induced metabolismrelated. metabolismrelated metabolism related. related metabolism-related

ABSTRACT Fat deposition is higher in fast growing chickens than in slow growing chickens. The liver is the major organ for lipogenesis and fat deposition in chickens, although genetic background, age, and gender also influence fat deposition. In the present study, we aimed to explore the molecular mechanisms underlying fat deposition in liver and abdominal fat. We determined the expression abundances of the key genes regulating fat metabolism in fast-growing (FG) broilers (Cobb) and slow-growing (SG) broilers (HS1) and found that ACC, FAS, PGC-1α, PPARγ, SREBP-1c and PLIN1genes were expressed in the abdominal fat and liver tissues of FG and SG. ANOVA analysis showed that the breed, age, and tissue factors influenced the expressions of ACC, FAS, PGC-1α, PPARγ, SREBP-1c, and PLIN1 genes in the liver and abdominal fat of FG and SG. Also, the expressions of PPARγ and PLIN1 in the liver of SG were higher than that of FG. The results suggest that the differences in adipocyte development and adipose deposition between breeds are due to genetic factors.

ABSTRACT Heparan sulfate proteoglycans (HSPGs) are present on the cell surface and in the extracellular matrix in all metazoans. HSPGs interact with growth factors and receptors through heparan sulfate (HS) chains. The sulfation pattern of heparan sulfate chains influences signaling events mediated by heparan sulfate proteoglycans located on the cell surface. SULF1 and SULF2 are two endo-sulfatases that can cleave specific 6-O-sulfate groups within the heparan chains. To determine their possible roles in tissues and satellite cells in vitro, their expression pattern was examined in tissues from 40-day-old chickens and in satellite cells from the breast muscles of 1-week-old and 2-week-old chickens using RT-PCR and immunocytochemistry analyses. The SULF1 and SULF2 transcripts were widely distributed in various tissues. Upon increasing culture times in chicken´s primary skeletal muscle satellite cells, SULF1 and SULF2 expression in 1-week-old chickens was significantly higher than in 2-week-old chickens, suggesting that sulfatases play a key role in satellite cell development. Therefore, our findings increase our knowledge of sulfatase expression diversity and provide a solid basis for further research concerning this molecular mechanism.

ABSTRACT The aim of this study was to investigate the effects of glutamine (Gln) on the intestinal mucosal structure and immune cells of broilers infected with Salmonella Enteritidis. 160 1-d-old commercial Arbor Acres (AA) broilers were randomly selected to receive one of four treatments, each of which had 5 replicates. Each replicate consisted of 8 chicks subjected to a 21-d feeding trial. Group I served as the unchallenged (CON). All birds in groups II (SCC) - IV were challenged with 2.0 × 104 CFU/mL of S. Enteritidis. The birds in groups III and IV were treated with 0.5% and 1.0% Gln. The results showed that S. Enteritidis infection led to a decrease in the relative length and weight, villus height:crypt depth (VH:CD) of the jejunum and ileum, the number of intraepithelial lymphocyte cells, and number of goblet cells and an increase in the number of mast goblet cells compared with the measurements of these parameters in the CON group (p<0.05). In addition, the Gln groups had increased relative length and weight, VH:CD of the jejunum and ileum, numbers of intraepithelial lymphocyte cells, and numbers of goblet cells and decreased crypt depth in the jejunum and ileum and numbers of mast goblet cells compared with the measurements of these parameters in the SCC group (p<0.05). It was concluded that Gln added to broiler diets can effectively alleviate the intestinal mucosal damage caused by S. Enteritidis infection and improve its normal defense barrier function.

ABSTRACT The melanocortin 1 receptor (MC1R) gene plays a key role in controlling the deposition of melanin. In mammals, the MC1Rgene is regarded as a major candidate gene in the control of melanin formation. In domestic animals, the MC1R gene mainly controls the expression of coat, skin, and plumage color in mammals and birds. In order to breed chickens with dark-green shank faster, we screened the molecular markers for shank color in a HS chicken population by exploring the relationship between polymorphism of the MC1R gene and three different shank colors (light green, dark green and yellow). Two primer pairs for code region of the MC1R gene were designed in the basic of chicken genomic sequence. DNA sequencing was performed to detect the polymorphisms and PCR was used to amplify DNA fragment. Sequences analysis indicated that 7 SNPs were predominant the three HS chicken populations with different shank color, including g.18,287,945C>T, g.18,288,088T>C, g.18,288,150G>A, g.18,288,303A>G, g.18,288,512G>A, g.18,288,513T>C, and g.18,288,520A>C. Association analysis revealed that the dark-green shank population showed moderate polymorphism, whereas the light-green shank population showed low polymorphism among overall 7 SNPs and that SNP6 (g.18,288,513T>C) may be significantly associated with three different shank colors in HS chickens. The haplotype CTGGACA had the largest haplotype frequencies, accounting for 56.22%, and the haplotype combination H1H1 is mainly distributed in the dark-green shank population, and may be used as molecular maker for marker-assisted selection of shank color in HS chickens.

ABSTRACT The present study was conducted on one-day-old broilers to investigate the effects of methionine deficiency on B lymphocytes and immunoglobulins (sIgA, IgA, IgG, IgM) in the cecal tonsil of Cobb broiler chicken. Methods including immunohistochemistry (IHC), western blot (WB), quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) and Enzyme-Linked Immuno Sorbent Assay (ELISA) were used, and the study lasted for 6 weeks. It was found that the IgA+ B lymphocytes mainly existed in the diffuse lymphoid tissues and lymphoid follicles of the cecal tonsil. The data indicated that the number of IgA+ B lymphocytes was decreased in the methionine deficiency group, and the WB and qRT-PCR results suggested that the protein and mRNA expression of CD19 were reduced in the methionine deficiency group. Besides, the contents of sIgA, IgA, IgG and IgM determined by ELISA decreased in the methionine deficiency group. It could be concluded that methionine deficiency exerts significantly negative effects on the humoral immune function of the intestinal mucosal immunity.

ABSTRACT There is increasing evidence that bone morphogenetic protein 6 (BMP6) plays critical roles in regulating various stages of ovarian follicle development in mammals. However, the mechanisms of regulation of BMP6 in the chicken ovary remain unclear. In this study, mRNA and protein expression level of BMP6 in chicken ovarian follicles at different development stages were determined by qRT-PCR and western blot separately. Different concentrations of BMP6 protein and FSH were added to the culture medium, and the effects to proliferation of granulose cells were detected, further effect on expression pattern of progesterone synthesis associated genes were also analyzed by qRT-PCR and Western blotting and the secretion of progesterone was detected by ELISA. The results showed that mRNA and protein expression level of BMP6 increased significantly in the follicle with the development of follicle (p<0.05) and reached a peak at F1 follicle. Adding concentration of 50ng/ml and 100ng/ml of BMP6 protein promoted significantly the proliferation of granulosa cells (p<0.05), as well as up-regulated the expression of Steroid hormone synthesis acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) genes in mRNA and protein level. Meanwhile, the secretion of progesterone was significantly higher in the group that added BMP6 and FSH separately than blank control group (p<0.05) and reached a peak in the group that both added BMP6 and FSH. Collectively, these findings highlight that BMP6 is associated with proliferation of follicular cells and the synthesis of progesterone, which indicated that it took an important role in the follicular development of chicken.

ABSTRACT In this study, a method utilizing PCR-restriction fragment length polymorphism (PCR-RFLP) of a mitochondrial gene was developed for the identification of chicken (Gallus gallus), quail (Coturnix coturnix), and common pigeon (Columba livia) meat. PCR products of ~440 bp were obtained from the 12S rRNA gene of these three birds using a pair of universal primers. The three terrestrial birds can be distinguished using one restriction endonuclease, Alu I, which was selected based on species-specific variations in the mt 12S rRNA gene sequence using 9 newly-obtained and 44 published chicken, quail and pigeon sequences. This method was also successfully used to identify commercial quail and pigeon meat products, which were found to be adulterated with chicken meat. Additionally, our method had relatively high sensitivity for detecting a meat mixture. Ten percent of chicken meat in the mixed quail and pigeon sample was detectable. This assay can be useful for the accurate identification of meats from terrestrial birds, avoiding mislabeling or fraudulent species substitution in meat products.

ABSTRACT Reticuloendotheliosis virus (REV) infection has frequently affected the poultry industry in recent years. The infection with REV weakens the immune responses of the infected poultry. It is reported that Enteromorphaclathrata polysaccharides are capable of regulating immune function. In order to investigate the immuno regulatory effects of Enteromorphaclathrata polysaccharides (EPS) on the response of REV-infected broilers to a live attenuated Newcastle disease (ND) vaccine. Broilers were intraperitoneally injected with REV at one day of age, subcutaneously infected with EPS at 2 days of age, and vaccinated by nasal drip with a live attenuated ND (Lasota strain) vaccine at 5 days of age. Immune organ index, secretory immunoglobulinA (SIgA), peripheral blood heterophil to lymphocyte ratios (H/L ratio), peripheral blood lymphocyte transformation rates, and interferon-gamma (IFN-γ) and interleukin-2 (IL-2) levels were measured at 3, 7, 14, 21, 28, 35, 42, and 56 days of age. The results showed that EPS increased the immune organ index, and the secretion of small intestine secretory immunoglobulin A, serum ND antibody titers, blood H/L ratio, peripheral blood lymphocyte transformation rates, and IL-2 and IFN-γ levels. These results indicate that EPS are able to enhance the immune responses of chickens both to REV infection and to ND vaccination. Therefore, Enteromorphaclathrata polysaccharides can be considered as an immune regulator in the future.

ABSTRACT BMP6, a member of the subfamilies of the morphogenetic proteins (BMPs), plays a crucial role in osteogenic and chondrocyte differentiation in vitro and stimulates chondrogenesis, making chondrocytes differen-tiate on their terminal stage. The objective of this study is to explore the relationship between polymorphism of BMP6 gene and slaughter traits in chicken respectively. We screened the exonic and intronic regions of BMP6 gene by DNA pool construction and amplified DNA fragment by PCR, and finally, we got nine SNPs. Association analysis revealed that BMP6 had no significant association among all slaughter traits in Yellow bantam chicken. However, BMP6 had a significant difference with femur weight, tibia weight, femur length (p<0.05), and was extremely significant with tibia length (p<0.01) in Avian chicken. Moreover, femur perimeter also had significant correlation with BMP6 in Avian chicken. These results provide useful information for further investigation on the function of chicken BMP6 gene.

OBJECTIVE: We attempted to evaluate maternal thyroid function in a new self-sequential longitudinal reference interval (SLRI) which we established recently. By this method, we analysed the correlation between pregnancy outcome, neonatal thyroid stimulating hormone (TSH) level and maternal thyroid diseases. METHODS: A total of 1744 pregnant women participated in the study and 1747 babies were born from those women (three bore twins). The levels of TSH, free thyroxine (FT4) and thyroid peroxidase antibodies (TPO-Ab) of mothers were quantified by electrochemistry immunoassay (ECL). The levels of neonatal blood TSH were detected by time-resolved fluorescence immunoassay (TRFIA). All data were collected and statistically analysed by SPSS 13.0 software. RESULTS: With our new SLRI method, we found that 0.11%~3.84% pregnant women would get thyroid diseases. Subclinical hypothyroidism was the most common maternal thyroid disorder. Being positive for thyroid peroxidase antibodies was a significant risk factor of subclinical hypothyroidism during pregnancy. The median, P2.5~P97.5, and interquartile range (IQR) of neonatal TSH (N-TSH) of 1747 babies were 2.72 mIU/L, 0.10~8.01 mIU/L and 2.62 mIU/L, respectively; 28.6% of pregnant women with thyroid diseases developed pregnancy complications. The prevalence was significantly higher than in the normal thyroid function group (p< 0.001). The levels of N-TSH were low correlated with maternal TSH levels (p < 0.05), but there were no significant correlations between N-TSH and maternal FT4 and maternal TPO-Ab (p > 0.05). CONCLUSIONS: Thyroid disorders, especially subclinical hypothyroidism, are common in pregnant women. These disorders are associated with pregnancy and fetal outcome. Routine maternal thyroid function screening is important and should be recommended.

OBJETIVO: Intentamos evaluar la función tiroidea materna en un nuevo intervalo de referencia longitudinal auto-secuencial (SLRI) que establecimos recientemente. Por este método, analizamos la correlación entre el resultado del embarazo, el nivel de la hormona estimulante de la tiroides (TSH) en neonatos, y las enfermedades tiroideas maternas MÉTODOS: Un total de 1744 mujeres embarazadas participó en el estudio y 1747 bebés nacieron de esas mujeres (tres de ellas tuvieron gemelos). Los niveles de TSH, la tiroxina libre (FT4), y los anticuerpos de la peroxidasa tiroidea (TPO-Ab) de las madres, fueron cuantificados mediante inmunoensayo electroquímico (ECL). Los niveles de TSH en la sangre de los neonatos, fueron determinados mediante inmunoensayo por fluorescencia resuelto en el tiempo (TRFIA). Todos los datos fueron recogidos y analizados estadísticamente usando el software SPSS 13.0 RESULTADOS: Con nuestro nuevo método SLRI, encontramos que 0.11%~3.84% de las mujeres embarazadas contraerán enfermedades tiroideas. El hipotiroidismo subclínico fue el trastorno de la tiroides materna más común. Ser positivo a los anticuerpos de la peroxidasa tiroidea fue un factor de riesgo significativo del hipotiroidismo subclínico durante el embarazo. La mediana, P2.5~P97.5, y el rango intercuartil (IQR) de la TSH (N-TSH) neonatal de los 1747 bebés fueron 2.72 mIU/L, 0.10~8.01 mIU/L y 2.62 mIU/L respectivamente. El 28.6% de las mujeres embarazadas que tenían enfermedades tiroideas, desarrollaron complicaciones del embarazo. La prevalencia fue significativamente más alta que en el grupo con función tiroidea normal (p < 0.001). Los niveles de N-TSH fueron bajos en correlación con los niveles de TSH maternos (p < 0.05), pero no hubo ninguna correlación significativa entre la N-TSH y la FT4 materna, y la TPO-Ab materna (p > 0.05). CONCLUSIÓNS: Los trastornos tiroideos, especialmente el hipotiroidismo, son comunes en las mujeres embarazadas.Estos trastornos se hallan asociados con el resultado del embarazo y el resultado fetal. El tamizaje de rutina de la función tiroidea materna es importante y debe recomendarse.