BackgroundThe availability of the bovine genome sequence and SNP panels has improved various genomic analyses, from exploring genetic diversity to aiding genetic selection. However, few of the SNP on the bovine chips are polymorphic in buffalo, therefore a panel of single nucleotide DNA markers exclusive for buffalo was necessary for molecular genetic analyses and to develop genomic selection approaches for water buffalo. The creation of a 90K SNP panel for river buffalo and testing in a genome wide association study for milk production is described here.MethodsThe genomes of 73 buffaloes of 4 different breeds were sequenced and aligned against the bovine genome, which facilitated the identification of 22 million of sequence variants among the buffalo genomes. Based on frequencies of variants within and among buffalo breeds, and their distribution across the genome, inferred from the bovine genome sequence, 90,000 putative single nucleotide polymorphisms were selected to create an Axiom® Buffalo Genotyping Array 90K.ResultsThis 90K “SNP-Chip” was tested in several river buffalo populations and found to have ∼70% high quality and polymorphic SNPs. Of the 90K SNPs about 24K were also found to be polymorphic in swamp buffalo. The SNP chip was used to investigate the structure of buffalo populations, and could distinguish buffalo from different farms. A Genome Wide Association Study identified genomic regions on 5 chromosomes putatively involved in milk production.ConclusionThe 90K buffalo SNP chip described here is suitable for the analysis of the genomes of river buffalo breeds, and could be used for genetic diversity studies and potentially as a starting point for genome-assisted selection programmes. This SNP Chip could also be used to analyse swamp buffalo, but many loci are not informative and creation of a revised SNP set specific for swamp buffalo would be advised.

Resveratrol is one of the most investigated natural polyphenolic compounds and is contained in more than 70 types of plants and in red wine. The widespread interest in this polyphenol derives from its antioxidant, anti-inflammatory and anti-aging properties. Several studies have established that resveratrol regulates animal reproduction. However, the mechanisms of action and the potential therapeutic effects are still unclear. This review aims to clarify the role of resveratrol in male and female reproductive functions, with a focus on animals of veterinary interest. In females, resveratrol has been considered as a phytoestrogen due to its capacity to modulate ovarian function and steroidogenesis via sirtuins, SIRT1 in particular. Resveratrol has also been used to enhance aged oocyte quality and as a gametes cryo-protectant with mainly antioxidant and anti-apoptotic effects. In males, resveratrol enhances testes function and spermatogenesis through activation of the AMPK pathway. Furthermore, resveratrol has been supplemented to semen extenders, improving the preservation of sperm quality. In conclusion, resveratrol has potentially beneficial effects for ameliorating ovarian and testes function.

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To increase the sustainability of trout farming, the industry requires alternatives to fish-based meals that do not compromise animal health and growth performances. To develop new feeds, detailed knowledge of intestinal morphology and physiology is required. We performed histological, histochemical, immunohistochemical and morphometric analysis at typical time points of in vivo feeding trials (50, 150 and 500 g). Only minor changes occurred during growth whereas differences characterized two compartments, not linearly distributed along the intestine. The first included the pyloric caeca, the basal part of the complex folds and the villi of the distal intestine. This was characterized by a significantly smaller number of goblet cells with smaller mucus vacuoles, higher proliferation and higher apoptotic rate but a smaller extension of fully differentiated epithelial cells and by the presence of numerous pinocytotic vacuolization. The second compartment was formed by the proximal intestine and the apical part of the posterior intestine complex folds. Here we observed more abundant goblet cells with bigger vacuoles, low proliferation rate, few round apoptotic cells, a more extended area of fully differentiated cells and no pinocytotic vacuoles. Our results suggest that rainbow trout intestine is physiologically arranged to mingle digestive and absorptive functions along its length.

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Simple SummaryGrazing activity is fundamental to natural grassland biodiversity preservation. Increasing summer aridity decreases the grassland pastoral value, negatively affecting animal morpho-functional features and production with detrimental effects on the extensive sheep farming sustainability. Since adipokines represent a link between a subject’s energy availability and tissue metabolism, we investigated the presence and distribution of the system composed of the adipokine apelin and its receptor in the mammary gland of sheep during the period between maximum flowering and maximum dryness of the pasture, providing a group of sheep with food supplementation. This work represents a part of a wider study aimed to buffer the negative effects of increasing summer drought stress on farm income and to maintain the grassland biodiversity. Our findings improve the knowledge of apelin/receptor system function on the sheep mammary gland and this could be a useful tool in the farm management practices.AbstractSheep are the most bred species in the Central Italy Apennine using the natural pastures as a trophic resource and grazing activity is fundamental to maintain the grassland biodiversity: this goal can be reached only ensuring an economical sustainability to the farmers. This study aimed to investigate the apelin/apelin receptor system in ovine mammary gland and to evaluate the differences induced by food supplementation, in order to shed light on this system function. A flock of 15 Comisana x Appenninica adult dry ewes were free to graze from June until pasture maximum flowering (MxF). From this period to pasture maximum dryness (MxD), in addition to grazing, the experimental group (Exp) was supplemented with 600 g/day/head of cereals. Apelin and apelin receptor were assessed by Real-Time PCR and immunohistochemistry on the mammary glands of subjects pertaining to MxF, MxD and Exp groups. They were detected in alveolar and ductal epithelial cells. The pasture maximum flowering group showed significant differences in apelin expression compared with experimental and MxD groups. Apelin receptor expression significantly differed among the three groups. The reduced apelin receptor expression and immunoreactivity levels during parenchyma involution enables us to hypothesize that apelin receptor plays a modulating role in the system control.

24Developmental competence determines the oocyte capacity to support initial embryo 25 growth, but the molecular mechanisms underlying this phenomenon are still ill-26 defined. Changes in microRNA (miRNA) expression pattern have been described 27 during follicular growth in several species. Therefore, aim of this study was to 28 investigate whether miRNA expression pattern in cow oocyte and follicular fluid (FF) 29 is associated with the acquisition of developmental competence. Samples were 30 collected from ovaries with more than, or fewer than, 10 mid-antral follicles (H-and 31 L-ovaries) because previous studies demonstrated that this parameter is a reliable 32 predictor of oocyte competence. After miRNA deep sequencing and bioinformatic 33 data analysis, we identified 58 miRNAs in FF and 6 in the oocyte that were 34 differentially expressed between H-and L-ovaries. Overall, our results indicate that 35 miRNA levels both in FF and in the ooplasm must remain within specific thresholds 36 and that changes in either direction compromise oocyte competence. Some of the 37 miRNAs found in FF (miR-769, miR-1343, miR-450a, miR-204, miR-1271 and miR-38 451) where already known to regulate follicle growth and their expression pattern 39 indicate that they are also involved in the acquisition of developmental competence. 40Some miRNAs were differentially expressed in both compartments but with opposite 41 patterns, suggesting that miRNAs do not flow freely between FF and oocyte. Gene 42Ontology analysis showed that the predicted gene targets of most differentially 43 expressed miRNAs are part of a few signalling pathways. Regulation of maternal 44 mRNA storage and mitochondrial activity seem to be the processes more 45 functionally relevant in determining oocyte quality. In conclusion, our data identified a 46 few miRNAs in the follicular fluid and in the ooplasm that modulate the oocyte 47Recently, microRNAs (miRNAs), which regulate gene expression at the mRNA level, 65 have been associated with folliculogenesis and oogenesis [10, 11]. MiRNAs, which 66 range in size from 18 to 25 nucleotides (nt), have been found in the different 67 compartments of ovarian follicles, including granulosa cells [12, 13], theca cells [14], 68 follicular fluid and the oocyte itself [15]. Studies on the role of miRNAs during follicle 69 development in humans [16][17][18], mice [19, 20], cattle [10, 21, 22], pigs [23] and 70horses [24] suggest that they regulate the cellular differentiation processes which 71 occur during follicular development. 72 Follicular fluid and germinal vesicle oocyte collection 111Ovaries were collected at a commercial abattoir and were transported to the 112 laboratory in warmed (27-30°C) Dulbecco Phosphate Buffered Saline (PBS). Ovaries 113 were classified into low and high antral follicle count categories according to the 114 methods used in previous works [28, 29]. Briefly, the ovaries were assigned to high 115 antral follicle count ovaries (H ovaries) when more than 10 mid-antral follicles (2-5 116 mm in dia...

We previously showed that, according to the frequency and distribution of specific cell types, the rainbow trout (RT) intestinal mucosa can be divided in two regions that form a complex nonlinear three-dimensional (3D) pattern and have a different renewal rate. This work had two aims. First, we investigated whether the unusual distribution of cell populations reflects a similar distribution of functional activities. To this end, we determined the protein expression pattern of three well-defined enterocytes functional markers: peptide transporter 1 (PepT1), sodium–glucose/galactose transporter 1 (SGLT-1), and fatty-acid-binding protein 2 (Fabp2). Second, we characterized the structure of RT intestinal stem-cell (ISC) niche and determined whether the different proliferative is accompanied by a different organization and/or extension of the stem-cell population. We studied the expression and localization of well-characterized mammal ISC markers: LGR5, HOPX, SOX9, NOTCH1, DLL1, and WNT3A. Our results indicate that morphological similarity is associated with similar function only between the first portion of the mid-intestine and the apical part of the complex folds in the second portion. Mammal ISC markers are all expressed in RT, but their localization is completely different, suggesting also substantial functional differences. Lastly, higher renewal rates are supported by a more abundant ISC population.