Effect of Dietary Amaranth (Amaranthus hybridus chlorostachys) Supplemented with Enzyme Blend on Egg Quality, Serum Biochemistry and Antioxidant Status in Laying Hens.

A feeding trial was performed to assess the effects of dietary raw amaranth (Amaranthus hybridus chlorostachys) grain (RAG), with or without an enzyme blend, on the productive performance, blood biochemistry, and antioxidant status in laying hens. The trial was conducted following a completely randomized design by factorial method, including five levels of RAG (0, 10, 20, 30, and 40%, respectively) and two levels of enzyme blend (0 -E and 0.025 +E %). A total of 960 White Leghorn (Hy-line W-36) laying hens (56 weeks of age) were divided into 10 groups with eight repetitions, including 12 birds. The trial period was ten weeks. Results showed that RAG levels in feed (>10%) led to a significant decrease in blood total cholesterol (TC), but they also significantly decreased feed conversion ratio (FCR) (p ˂ 0.05) as measured by feed intake (FI), hen daily production (HDP), egg weight (EW), and mass (EM), leading to overall worse productivity compared to the control group. On the contrary, the addition of the enzyme blend led to an improvement in the investigated production traits (p ˂ 0.05), with the exception of HDP. The enzyme blend was also capable of recovering productive performance when combined with low concentrations of RAG (10%) (p ˂ 0.05), and RAG × enzyme blend groups showed the lowest values of TC (p ˂ 0.05). Moreover, the interaction effects for atherogenic index (LDL/HDL) indicated a significant and promising reduction in response to the addition of RAG both in the presence and absence of the enzyme blend (p ˂ 0.05), and this additive also significantly reduced levels of egg yolk cholesterol (p ˂ 0.05). In summary, the evidence gathered in this trial showed that dietary RAG had positive effects on egg quality characteristics, leading to the production of low-cholesterol eggs, and, at the same time, it may improve the health status of laying hens. Furthermore, the addition of an enzyme blend allowed feeding up to 10% RAG in the diet, leading to an optimal balance between animal productivity and the beneficial effects of RAG.

Supplementation of broiler chicken diets with bovine lactoferrin improves growth performance, histological parameters of jejunum and immune-related gene expression.

The aim of this study was to investigate the influence of dietary supplementation of bovine lactoferrin (bLF) on growth performance, carcass traits, histomorphology of jejunum, immune function and hepatic and splenic gene expression of interferon-gamma (IFN-γ) and interleukine-2 (IL-2) in broiler chickens. A total of 240 one-day-old Ross 308 male broiler chickens were randomly allotted into six dietary treatments with four replicate pens (10 chicks per pen) and fed experimental diet in 3 feeding phases (starter: d 0-10, grower: d 11-24 and finisher: d 25-42). The experimental treatments were (1) corn-soya bean meal-based basal diet (control), (2-5) basal diet supplemented with 200, 400, 600, 800 mg/kg bLF, respectively, and (6) basal diet supplemented with 200 mg/kg oxytetracycline (OTC). The average body weight gain (ABWG) of broilers fed 800 mg/kg bLF was 8.48% higher than those fed a corn-soybean meal-based diet during the starter period (d 0-10) (linear effect, p = 0.002; quadratic effect, p = 0.24). Average daily feed intake (ADFI) and the feed conversion ratio (FCR) were not affected (p>0.05) by bLF supplementation. At 42 days of age, the breast meat percentage and carcass yield of broilers fed 800 mg/kg bLF compared with the control group significantly increased by 9.51% and 6.03% respectively (p < 0.05). Compared with the chicks fed the control diet, the chicks fed diets supplemented with bLF had higher villus height, muscle thickness and villus surface area (p > 0.05). Dietary bLF inclusion increased the total immunoglobulin (IgT) titre against sheep red blood cells (SRBCs) antigen (linear effect, p = 0.031; quadratic effect, p = 0.035) and improved the phytohaemagglutinin-P (PHA-P)-skin test of broilers. Compared with the control, bLF enhanced the gene expression of IFN-γ in spleen (p = 0.048, linear effect, p = 0.009; quadratic effect, p = 0.093) and liver (p = 0.012, linear effect, p = 0.008; quadratic effect, p = 0.01) and IL-2 expression in spleen (p = 0.021, linear effect, p = 0.026; quadratic effect, p = 0.103). The bLF supplementation had no effect on IL-2 gene expression in liver (p > 0.05, linear effect, p = 0.213; quadratic effect, p = 0.159). In conclusion, we found that supplementation of broiler diets with 800 mg/kg bLF can improve the growth performance, carcass yield, cell-mediated and antibody-mediated immune responses and enhance the IL-2 and IFN-γ gene expression of broilers.

Investigation of chicken housekeeping genes using next-generation sequencing data.

Accurate normalization of the gene expression assays, using housekeeping genes (HKGs), is critically necessary. To do so, selection of a proper set of HKGs for a specific experiment is of great importance. Despite many studies, there is no consensus about the suitable set of HKGs for implementing in the quantitative real-time PCR analyses of chicken tissues. A limited number of HKGs have been widely used. However, wide utilization of a little number of HKGs for all tissues is challenging. The emergence of high-throughput gene expression RNA-seq data has enabled the simultaneous comparison of the stability of multiple HKGs. Therefore, employing the average coefficient of variations of at least three datasets per tissue, we sorted all reliably expressed genes (REGs; with FPKM ≥ 1 in at least one sample) and introduced the top 10 most suitable and stable reference genes for each of the 16 chicken tissues. We evaluated the consistency of the results of five tissues using the same methodology on other datasets. Furthermore, we assessed 96 previously widely used HKGs (WU-HKGs) in order to challenge the accuracy of the previous studies. The New Tuxedo software suite was used for the main analyses. The results revealed novel, different sets of reference genes for each of the tissues with 17 common genes among the top 10 genes lists of 16 tissues. The results did disprove the suitability of WU-HKGs such as Actb, Ldha, Scd, B2m, and Hprt1 for any of the tissues examined. On the contrary, a total of 6, 13, 14, 23, and 32 validated housekeeping genes (V-HKGs) were discovered as the most stable and suitable reference genes for muscle, spleen, liver, heart, and kidney tissues, respectively. Although we identified a few new HKGs usable for multiple tissues, the selection of suitable HKGs is required to be tissue specific. The newly introduced reference genes from the present study, despite lacking experimental validation, will be able to contribute to the more accurate normalization for future expression analysis of chicken genes.

Bayesian threshold analysis of litter size in sheep

Background: Litter size at birth (LSB) is one of the most important economic traits in sheep and could be used in genetic improvement schemes for meat production. LSB is inherently a categorical trait and should be analysed with threshold models. Objective: Bayesian threshold models were used to analyze sheep LSB to estimate genetic parameters. Methods: Data was based on 7,901 LSB records from 14,968 dams and 682 sires collected from 1986 to 2012 at Makouie Sheep Breeding Station in Iran. Means of posterior distributions (MPDs) of LSB's genetic parameters were estimated, and the best-fitted models were selected using the deviance information criterion. Results: In the repeated measurement analysis, the estimated direct and maternal heritabilities, and permanent environmental effect (±SE), according to the best-fitted model (model 5), were 0.01 (0.010), 0.02 (0.014), and 0.01 (0.011), respectively. In the univariate analysis, the best estimates of direct and maternal heritabilities were 0.12 (0.064) and 0.08 (0.045), respectively. An increasing trend for direct and maternal heritabilities was observed in parity 2 (0.15 (0.082) and 0.25 (0.083), respectively). In the bivariate analysis, the best estimates of direct and maternal heritabilities for LSB were 0.03 (0.027) and 0.22 (0.041), respectively. The direct and maternal genetic correlations among parities were 0.25 (0.054) and 0.12 (0.021), respectively. Conclusions: The results showed a considerable influence of environmental factors on LSB in each parity of sheep; also, statistically different genetic parameters (p<0.05) were obtained from one parity to another, indicating the different and large influences of genetic and environmental factors for each parity.

Antecedentes: El tamaño de la camada al nacer (LSB) es inherentemente un rasgo categórico y debe analizarse con modelos de umbral. El LSB es uno de los rasgos de producción de carne más importantes en las ovejas y podría usarse en esquemas de mejora genética para la producción de carne. Objetivo: Se utilizaron modelos de umbral bayesiano para analizar el tamaño de la camada de ovejas al nacer (LSB) y estimar parámetros genéticos. Métodos: Los datos se basaron en 7.901 registros de LSB de 14.968 ovejas y 682 carneros recolectados de 1986 a 2012 en la estación de cría de ovejas Makouie en Irán. Se estimaron las medias de distribuciones posteriores (MPD) de los parámetros genéticos de LSB y se seleccionaron los modelos mejor ajustados utilizando el criterio de información de desviación. Resultados: En los análisis de medición repetida, la heredabilidad materna y directa estimada y el efecto ambiental permanente (±SE), según el modelo mejor ajustado (modelo 5), fueron 0,01 (0,010), 0,02 (0,014) y 0,01 (0,011), respectivamente. En el análisis univariado, las mejores estimaciones de heredabilidad directa y materna fueron 0,12 (0,064) y 0,08 (0,045), respectivamente. Se observó una tendencia creciente de heredabilidades directas y maternas en la paridad 2 (0,15 (0,082) y 0,25 (0,083), respectivamente). En el análisis bivariado, las mejores estimaciones de heredabilidad directa y materna para LSB fueron 0,03 (0,027) y 0,22 (0,041), respectivamente. Las correlaciones genéticas directas y maternas entre partos fueron 0,25 (0,054) y 0,12 (0,021), respectivamente. Conclusiones: Los resultados mostraron una influencia considerable de los factores ambientales sobre el LSB en cada parto de las ovejas; además, se obtuvieron parámetros genéticos estadísticamente diferentes (p<0.05) de un parto a otro, indicando las diferentes y grandes influencias de factores genéticos y ambientales para cada parto en ovejas. Los resultados de este estudio se pueden precisar aún más utilizando datos de SNP de todo el genoma sobre diferentes partes para manejar una amplia gama de problemas relacionados con la interacción del entorno genético del rasgo LSB.

Antecedentes: O tamanho da ninhada ao nascer (LSB) é inerentemente uma característica categórica e deve ser analisada com modelos de limiar. LSB é uma das características mais importantes de produção de carne em ovinos e pode ser usado em esquemas de melhoramento genético para a produção de carne. Objetivo: Modelos de limiar bayesiano foram usados para analisar o tamanho da ninhada de ovelhas ao nascer (LSB) para estimar parâmetros genéticos. Métodos: Os dados foram baseados em 7.901 registros LSB de 14.968 ovelhas e 682 carneiros coletados de 1986 a 2012 na Estação de Criação de Ovinos Makouie no Irã. Médias de distribuições posteriores (MPDs) dos parâmetros genéticos de LSB foram estimadas e os modelos mais bem ajustados foram selecionados usando o critério de informação de desvio. Resultados: Nas análises de medidas repetidas, as herdabilidades diretas e maternas estimadas e o efeito do ambiente permanente (±SE), de acordo com o modelo mais bem ajustado (modelo 5), foram 0,01 (0,010), 0,02 (0,014) e 0,01 (0,011), respectivamente. Na análise univariada, as melhores estimativas das herdabilidades direta e materna foram 0,12 (0,064) e 0,08 (0,045), respectivamente. Uma tendência crescente para as herdabilidades direta e materna foi observada na paridade 2 (0,15 (0,082) e 0,25 (0,083), respectivamente). Na análise bivariada, as melhores estimativas de herdabilidades direta e materna para LSB foram 0,03 (0,027) e 0,22 (0,041), respectivamente. As correlações genéticas diretas e maternas entre os partos foram 0,25 (0,054) e 0,12 (0,021), respectivamente. Conclusões: Os resultados mostraram uma influência considerável dos fatores ambientais na LSB em cada paridade de ovelhas; também, parâmetros genéticos estatisticamente diferentes (p<0,05) foram obtidos de uma paridade para outra, indicando as diferentes e grandes influências de fatores genéticos e ambientais para cada paridade em ovinos. Os resultados deste estudo podem ser ainda mais definidos usando dados SNPs de todo o genoma em diferentes partes para lidar com uma ampla gama de problemas relacionados à interação do ambiente genético do traço LSB.

Single-nucleotide polymorphism detecting of some candidate genes related to lipid metabolism in Booroola Merino-Afshari sheep by Bayesian model averaging.

This study was designed to identify single-nucleotide polymorphisms (SNPs) of some candidate genes related to lipid metabolism and their association with carcass fat in male crossbred lambs. Hence, 96 of almost 11-month-old Booroola Merino-Afshari crossbred lambs (first-generation backcross) were used by considering their phenotypic carcass traits. Then, DNA was extracted and DNA targets were amplified using designed specific primers by PCR procedure. Identification of potential SNPs was done by a direct sequencing method for LEP, FABP4, DGAT1, GH, and TRIB3 genes using the sequencing-RFLP procedure. Then, the most probable statistical models based on additive and genotypic effects of identified SNPs in each trait were obtained by the Bayesian model averaging (BMA) approach of R software (Ver. 3.3.1) to assess the association of SNPs with traits. Detected SNPs in this study included two SNPs in exon 3 of LEP, one SNP in exon 2 of TRIB3, one SNP in intron 2 of FABP4, one SNP in 5' UTR of DGAT1, and two SNPs in 3' UTR of GH genes. For carcass weight trait, one of the identified SNP genotypes in the LEP (c.587G > A) had a higher probability in the model. Carcass weight of lambs with GA genotype was 2.46 kg heavier than GG genotype. Also, two genes of TRIB3 and GH2 had the highest probability in the models of fat tail and waste weight, respectively. Based on the results, these polymorphisms can be used in the marker-assisted selection of breeding programs and designing DNA chips for genomic selection.

Modeling lactation curves and estimation of genetic parameters in Holstein cows using multiple-trait random regression models.

We compared the goodness of fit of three mathematical functions (including: Legendre polynomials, Lidauer-Mäntysaari function and Wilmink function) for describing the lactation curve of primiparous Iranian Holstein cows by using multiple-trait random regression models (MT-RRM). Lactational submodels provided the largest daily additive genetic (AG) and permanent environmental (PE) variance estimates at the end and at the onset of lactation, respectively, as well as low genetic correlations between peripheral test-day records. For all models, heritability estimates were highest at the end of lactation (245 to 305 days) and ranged from 0.05 to 0.26, 0.03 to 0.12 and 0.04 to 0.24 for milk, fat and protein yields, respectively. Generally, the genetic correlations between traits depend on how far apart they are or whether they are on the same day in any two traits. On average, genetic correlations between milk and fat were the lowest and those between fat and protein were intermediate, while those between milk and protein were the highest. Results from all criteria (Akaike's and Schwarz's Bayesian information criterion, and -2*logarithm of the likelihood function) suggested that a model with 2 and 5 coefficients of Legendre polynomials for AG and PE effects, respectively, was the most adequate for fitting the data.

Evaluation of the Effect of miR-26b Up-Regulation on HbF Expression in Erythroleukemic K-562 Cell Line.

BACKGROUND: The major hemoglobin in the fetus is hemoglobin F (HbF) (α2γ2), whereas in adult humans, hemoglobin A (α2ß2) is predominately expressed. Several studies have indicated that expression of the HbF subunit γ-globin might be regulated post-transcriptionally. This could be done by small non-coding RNAs called microRNAs which target mRNAs in a sequence-specific manner and lead to translational repression or mRNA decay. The aim of this study is to evaluate the effect of miR-26b up-regulation on γ-globin gene expression in K-562 cell line. METHODS: These cells were grown in RPMI 1640 and pre miR-26b and were transfected within K-562 cell line using lentiviral vector. After RNA extraction and cDNA synthesis in selected days, miRNA up-regulation was confirmed by miRNA real time PCR and then γand ßchain and GATA-1 expression were investigated by RT and QRT-PCR. RESULTS: The viability of cells before transfection was 90%. Three and 7 days after transfection, through the use of relative Q-PCR, the γ chain expression increased 3.7, 6.8 and 3.8 folds and GATA-1 expression increased 2.1, 6.0 and 8.0 in comparison with untransfected cells. CONCLUSION: The data suggest that miR-26b can be involved in the increase of γ-globin gene expression in K-562 cell line. We suggest that miR-26b may be a significant therapeutic target for increasing HbF levels in patients with sickle cell disease and ß-thalassemia.

The Effect of miR-210 Up-regulation on Proliferation and Survival of Mouse Bone Marrow Derived Mesenchymal Stem Cell.

BACKGROUND: Bone marrow derived mesenchymal stem cells (MSCs) are a population of multipotent progenitors which have the capacity of proliferation and differentiation into mesenchymal lineage cells. Hypoxia could promote the proliferation of MSCs. Micro-RNAs are endogenous RNAs that can play an important role in some processes such as proliferation and differentiation. MiR-210 could help for better proliferation of MSCs since this miRNA could activate HIF pathway. In current study we investigated if MSCs can preserve their differentiation and proliferation ability under normoxic conditions by upregulation of miR-210. MATERIALS AND METHODS: MSCs isolated from C57 BL/6 mice by flushing it's femurs into the cell culture media. After 72 hours, MSCs which are plastic adherent cells were attached to the flask and non-adherent cells were removed. Subsequently, MSCs induced to differentiate into osteocytes and adipocytes with specific differentiation media in order to confirm their identity and multipotency. Then miR-210 was inserted in Lentiviruse vectors and affected MSCs. In each passage, the number and viability of cells were evaluated. RESULTS: Comparison between miR-210 infected MSCs with control cells showed that miR-210 has ability to increase proliferation of MSCs significantly. CONCLUSION: We showed that miR-210 has ability to induce proliferation of MSCs without any negative effect on their differentiation abilities. Further studies are needed for evaluation of probable effects of miR-210 mechanisms on MSCs proliferation.

Chemical composition and the nutritive value of pistachio epicarp (in situ degradation and in vitro gas production techniques).

The nutritive value of pistachio epicarp (PE) was evaluated by in situ and in vitro techniques. Chemical analysis indicated that PE was high in crude protein (11.30%) and low in neutral detergent fiber (26.20%). Total phenols, total tannins, condensed tannins and hydrolysable tannins contents in PE were 8.29%, 4.48%, 0.49% and 3.79%, respectively. Ruminal dry matter and crude protein degradation after 48 hr incubation were 75.21% and 82.52%, respectively. The gas production volume at 48 hr for PE was 122.47 mL g(-1)DM. As a whole, adding polyethylene glycol (PEG) to PE increased (p < 0.05) gas production volumes, organic matter digestibility and the metabolizable energy that illustrated inhibitory effect of phenolics on rumen microbial fermentation and the positive influence of PEG on digestion PE. The results showed that PE possessed potentials to being used as feed supplements.