Transcriptome analysis and identification of genes associated with individual fertilisation rate differences in hen infundibulum.

1. Fertilisation rate is closely related to the reproductive performance and economic status of chicken laying breeders. In this study, two flocks (n = 1,029 in population I and n = 358 in population II) in the later laying period were used for investigating the individual differences among population fertilisation rates (FRs).2. The funnel and distal parts of the infundibulum were collected from nine individuals (five with low FR and four with high FR from population II) and RNA-sequencing (RNA-seq) method was used to investigate the transcriptome differences in fertilisation. Differences in fertilisation regulation were investigated by comparing the different parts (funnel and distal) of the infundibulum between the low FR and high FR groups.3. There were notable individual FR differences in both categories. Some individuals had a relative high FR (≥90%) for a long time (>5 days after AI), contrarily, some individuals lost the ability to fertilise eggs in a very short time.4. Differentially expressed genes (DEGs), such as DUSP7, PPP1R3B, FYB, OVA, OVALX and OVALY may be associated with sperm functional regulation, fertilisation and early-stage fertilised ovum development processes. DEGs such as AVBD1, AVBD2, AVBD6, NFATC2 and BANK1 indicated a severe immune response to sperm survival and fertilisation in the oviduct.5. The results suggested that individual differences should be considered in the breeding and reproduction process. The DEGs identified in this study may promote our understanding of different fertilisation regulation in laying breeders.

Effects of dietary substitution of peanut meal for soybean meal on egg production, egg quality, oxidative status, and yolk fatty acid profile in laying ducks.

There is an urgent need to evaluate and introduce alternatives for the unsustainable and traditional feed stuffs in poultry. This study evaluated the effects of graded replacement of soybean meal (SBM) by peanut meal (PNM) on egg production, egg quality, oxidative status, and yolk fatty acid profile in laying ducks. In total, 360 Longyan ducks aged 21 weeks were allocated to five treatments, each containing six replicates of 12 birds. Birds were fed the diets containing PNM replacing 0 (control), 25, 50, 75 or 100% of SBM for 16 weeks. With the increase of PNM level, egg production was improved (quadratic, P < 0.05), egg weight and feed consumption were decreased, feed conversion ratio and egg mass were impaired at 100% PNM, and yolk colour was enhanced (quadratic, P < 0.05). Plasma malondialdehyde concentration was increased at 100% PNM, while plasma glutathione concentration was decreased (quadratic, P < 0.01) as PNM substitution increased. Total cholesterol content in yolk decreased (P < 0.01) in response to increased PNM substitution. The contents of saturated fatty acids C20:0 and C22:0 in yolk increased (linear or quadratic, P < 0.05) with increased PNM substitution, but the contents of saturated fatty acid C18:0, monounsaturated fatty acid C22:1, and polyunsaturated fatty acids C18:3n-3 and C22:6n-3 in yolk were decreased (linear, P < 0.05) as PNM replacement increased. The contents in yolk of polyunsaturated fatty acid C18:2n-6, C20:2n-6, C20:3n-6, C20:4n-6, and total amount of polyunsaturated fatty acids increased with the highest contents obtained at 75% PNM substitution. The ratio between n-6 and n-3 increased (linear, P < 0.05) as PNM substitution increased. The transcript abundance in liver of peroxisome proliferators-activated receptors γ and fatty acid synthase showed quadratic (P < 0.05) responses with PNM replacement, with the highest expression of both genes being obtained with 75% substitution with PNM. Replacement of dietary SBM with 100% PNM decreased egg production and antioxidant capacity and increased the ratio of omega fatty acid in yolk between n-6 and n-3 in laying ducks. The obtained results indicate that PNM can be used to replace up to 75% of SBM in the diet of laying ducks without negative effects on the egg-laying production or egg quality. The regression model indicated that the maximal egg mass was obtained at no more than 67.6% replacement of SBM with PNM in the diet of laying ducks.

Effects of dietary inclusion of cassava starch-extraction-residue meal on egg production, egg quality, oxidative status, and yolk fatty acid profile in laying ducks.

This study was designed to evaluate the effects of different dietary levels of cassava starch extraction residue meal (CReM) on egg production, egg quality, oxidative status, egg yolk fatty acid profile, and hepatic expression of fatty acid metabolism-related genes. In total, 288 Longyan laying ducks aged 21 wk with similar BW were randomly assigned to 4 dietary treatments, each consisting of 6 replicates of 12 birds. The birds were fed a typical corn-soybean meal diet, which contained 0% (control), 5%, 10%, and 15% CReM, mainly replacing wheat bran, and the experiment lasted for 16 wk. The tested CReM levels did not show significant effects on the egg production, nonmarketable egg percentage, egg weight, daily egg mass, and FCR (g feed: g egg), but daily feed intake was reduced with increased CReM level (linear P < 0.001, quadratic P < 0.05). Yolk color increased (linear and quadratic, P < 0.01) with the increase in CReM level, but the Haugh unit, yolk proportion, albumen proportion, shell proportion, eggshell thickness, and eggshell strength were unaffected. Yolk contents of C11:0 and C12:0 (linear, quadratic, P < 0.01) and total saturated fatty acids increased, and the C22:1 level decreased (linear P < 0.01, quadratic P < 0.05) with the increase in CReM level, but the total monounsaturated fatty acids, the individual and total polyunsaturated fatty acids and n-6 and n-3 fatty acids, triglycerides, and total cholesterol in egg yolk were not affected. Hepatic gene expression revealed a significant increase in peroxisome proliferators-activated receptors γ (linear, quadratic, P < 0.001), but the expression of fatty acid synthase, sterol regulatory element binding protein 1 and apolipoprotein A1 genes were unaffected by CReM level. In conclusion, the results of the current study indicated that the CReM could be included up to 15% in laying duck diets without negative effects on the egg-laying rate, egg quality, and oxidative status. Dietary inclusion of CReM increased the yolk content of total saturated fatty acids and SOD activity in the liver.

Testicular transcriptome analysis under the dietary inclusion of l-carnitine reveals potential key genes associated with oxidative defense and the semen quality factor in aging roosters.

l-carnitine (LC) has been widely studied as a natural antioxidant molecule for treating low-fertility gametes. However, the molecular mechanism of the effect of LC supplementation on the testes of aging cocks has not been evaluated. Therefore, the objective of this study was to reveal the mechanism of testicular oxidative defense induced by LC supplementation in relation to the semen quality factor (SQF) in the aging cock through a transcriptome study conducted from a new perspective. A total of 24 Jinghong cocks at 55 wk of age were randomly divided into 2 groups (n = 12). All cocks were fed a standard isocaloric and isonitrogenic breeder diet (control, LC-0), and the other group was supplemented with LC at 150 mg/kg/d (treated, LC-150) for 12 wk. Remarkably, seminal characteristics and enzymes, sex hormones, and cock fertility related to testicular oxidative defense were considerably improved by LC supplementation. LC-150 testes showed the differential upregulation and downregulation of 97 and 90 transcripts, respectively, compared with LC-0 testes. Most upregulated transcripts were involved in testicular oxidative defense and spermiogenesis optimization, whereas the downregulated genes were responsible for oxidative stress, in contrast to the SQF. Functional analysis of the transcriptionally altered genes indicated the testicular deregulation of long-chain fatty acid and lipid peroxidation, enhancing fatty acid breakdown to release ATP production via ß-oxidation. These findings could lay the foundation for the discovery of new molecular markers of SQF-associated LC supplementation and potential targets for therapeutic intervention to optimize fertility in aging cocks.

The capability of coenzyme Q10 to enhance heat tolerance in male rabbits: evidence from improved semen quality factor (SQF), testicular oxidative defense, and expression of testicular melatonin receptor MT1.

Heat stress (HS) has a great influence on the etiology of male infertility. Coenzyme Q10 (CoQ10), known to have powerful antioxidant effects, has been reported to have such actions that are effective to treat infertility caused by HS. The aim of the present study was to investigate the antioxidative effect of CoQ10 on sperm quality, testicular antioxidant activities, and male fertility under HS. For this purpose, 18 mature male rabbits (aged 22 wk) of the Sinai Gabali breed were equally divided into 3 groups and placed at temperature-humidity index of 29 for 8 wk at a farm. The supplementation of CoQ10 at 0, 10, and 20 mg/kg of body weight was done in the first, second, and third groups, respectively. The results showed that the supplementation of CoQ10 had significant (P < 0.05) effect on semen quality factor (SQF) and testicular antioxidant activities by the supplementation of CoQ10. Moreover, a significant improvement in the concentration of testosterone, integrity of testicular DNA, and the expression of melatonin receptors was also observed, which were consistent with a significant improvement in buck fertility. The prolificacy was significantly increased (P < 0.05) in females when inseminated from bucks that were treated with CoQ10. Our results suggest that CoQ10 tends to decrease oxidative stress by enhancing testicular antioxidant activities, which are considered the most important factors for a buck's fertility. Hence, CoQ10 could be a suitable feed supplement to increase fertility, through enhancing the semen quality, in male rabbits and reducing the harmful effects of HS.

Faecal microbiome sequences in relation to the egg-laying performance of hens using amplicon-based metagenomic association analysis.

Exploring the composition and structure of the faecal microbial community improves the understanding of the role of the gut microbiota in the gastrointestinal function and the egg-laying performance of hens. Therefore, detection of hen-microbial interactions can explore a new breeding marker for the selection of egg production due to the important role of the gut microbiome in the host's metabolism and health. Recently, the gut microbiota has been recognised as a regulator of host performance, which has led to investigations of the productive effects of changes in the faecal microbiome in various animals. In the present study, a metagenomics analysis was applied to characterise the composition and structural diversity of faecal microbial communities under two selections of egg-laying performance, high (H, n = 30) and low (L, n = 30), using 16S rRNA-based metagenomic association analysis. The most abundant bacterial compositions were estimated based on the operational classification units among samples and between the groups from metagenomic data sets. The results indicated that Firmicutes phylum has higher significant (P < 0.01) in the H group than in the L group. In addition, higher relative abundance phyla of Bacteroides and Fusobacteria were estimated in the H group than the L group, contrasting the phyla of Actinobacteria, Cyanobacteria and Proteobacteria were more relative abundance in the L group. The families (Lactobacillus, Bifidobacterium, Acinetobacter, Flavobacteriaceae, Lachnoclostridum and Rhodococcus) were more abundant in the H group based on the comparison between the H and L groups. Meanwhile, three types of phyla (Proteobacteria, Actinobacteria and Cyanobacteria) and six families (Acinetobacter, Avibacterium, Clostridium, Corynebacterium, Helicobacter and Peptoclostridium) were more abundant in the L group (P < 0.01). Overall, the selection of genotypes has enriched a relationship between the gut microbiota and the egg-laying performance. These findings suggest that the faecal microbiomes of chickens with high egg-laying performance have more diverse activities than those of chickens with low egg-laying performance, which may be related to the metabolism and health of the host and egg production variation.

Bilateral apex pubis distance：a novel index for follicular development and egg laying status in domestic hens (Gallus gallus domesticus).

1. Measures of the bilateral apex pubis (BAP) in White Leghorn (WLH) hens aged from 16 to 64 weeks (from the time ovaries were completely undeveloped to senility) were ranked into various groups so the relationship between external traits (BAP distance, comb length, comb height, wattle length, wattle height, body weight) and internal traits (largest ovarian follicle diameter (LFD) and oviduct weight and length) could be determined.2. Using correlation and regression analysis with data from 16 weeks of age to the onset of lay, measurements of the apex pubis showed a strong correlation with LFD (r = 0.80, P < 0.01) and oviduct weight (r = 0.80, P < 0.01).3. Multiple comparisons among WLH hens from 16 to 43 weeks of age showed that the BAP distance increased in all groups ranked by LFD or by laying period.4. In the late laying period (>60 weeks of age), BAP distance showed a significant decrease (P < 0.01) in long-term non-laying hens compared to laying hens.5. These findings suggest BAP distance can serve as an index for evaluating the onset and probable stages of follicular development during sexual maturation and/or aid in the identification and selection of non-laying hens during reproduction.

Nutritional impacts of dietary oregano and Enviva essential oils on the performance, gut microbiota and blood biochemicals of growing ducks.

Nowadays, there is much legislation in the world devoted to restrict the use of synthetic antibiotics in the poultry industry, which could reduce performance rate and production profits. Various phyto-biotic growth promoters have been proposed to serve as antibiotic alternatives with emphasis on plant extracts and essential oils. This study was conducted to assess the impacts of using the oregano essential oil (OEO) (comprised of 5% thymol and 65% carvacrol) and Enviva essential oil (EEO) (4.5% cinnamaldehyde and 13.5% thymol) as phytobiotic feed additives (PFA) on growth performance, cecal microbiota and serum biochemicals of growing ducks. In total, 800 11-day-old ducklings, housed in 20 floor pens, were allotted randomly into five dietary treatments: (i) A basal diet (BD) (control), (ii.) BD+50 mg EEO/kg, (iii.) BD+100 mg EEO/kg, (iv.) BD+150 mg OEO/kg and (v.) BD+300 mg OEO/kg diet. The growth performance traits were studied between 11 and 42 days of age. At the experiment end, 40 ducks were slaughtered (eight/ treatment) and cecal digesta and blood samples were collected to estimate the cecal bacterial populations and serum blood biochemicals. The results indicated that the tested levels of OEO and EEO did not display any significant effect (P>0.05) on the duck's final BW, BW gain, growth rate, feed intake, feed conversion ratio or survivability rate. Besides, the different levels of EEO and OEO decreased the cecal populations of Coliforms (P<0.01), total aerobes (P<0.01) and lactose-negative Enterobacteria (P<0.05) in comparison with those of the control group. Finally, the tested EEO and OEO levels did not show any significant effect on the serum variables; in terms of total protein, albumin, globulin, total cholesterol, triglycerides, alanine aminotransferase and aspartate aminotransferase. In conclusion, the antimicrobial effect of the OEO and EEO against the cecal microbiota has been proven, while they did not display significant effects on the growth performance or blood variables of growing ducks.