The effects of feeding supplementation on the nutritional quality of milk and cheese from sheep grazing on dry pasture.

The effect of feeding supplementation on the nutritional characteristics of milk and cheese was studied in dairy sheep grazing on low mountain dry-grasslands during summer in typical sub-Mediterranean conditions of aridity. The control group (CG) of 25 sheep grazed on grass, while the experimental group (EG) of 25 sheep grazed on grass and received 600 g a day of a barley and corn mixture. Daily milk production showed a less pronounced decrease in EG than in CG (p ˂ 0.0368). After one month of supplementation, the concentrations of retinol and α-tocopherol in milk and cheese from EG were higher than CG (p < 0.05). Supplementary feeding had a positive effect on the fatty acid composition of the sheep milk. For the first time, positive effects on the volatile composition were found in EG cheese, displaying lower percentages of carboxylic acids associated with the pungent and rancid odour with respect to CG.

Dietary supplementation with olive mill wastewaters induces modifications on chicken jejunum epithelial cell transcriptome and modulates jejunum morphology.

BACKGROUND: The Mediterranean diet is considered one of the healthier food habits and olive oil is one of its key components. Olive oil polyphenols are known to induce beneficial effects in several pathological conditions, such as inflammatory bowel disease, and to contrast the proliferation of cancer cells or hypercholesterolemia. Polyphenols are also present in waste products derived from the olive industry: olive mill wastewaters (OMWW) are rich in polyphenols and there is an increasing interest in using OMWW in animal nutrition. OMWW are attributed with positive effects in promoting chicken performance and the quality of food-derived products. However, a tissue-specific transcriptome target analysis of chickens fed with OMWW has never been attempted. RESULTS: We explored the effect of dietary OMWW on the intestinal function in broilers. A morphological analysis of the jejunum revealed that OMWW reduced crypt depth, whereas no significant modifications were observed for villus height and the villus height/crypt depth ratio. An RNA Sequencing analysis was performed on isolated, intestinal, epithelial cells and 280 differentially expressed genes were found using a count-based approach. An enrichment analysis revealed that the majority of up regulated genes in the OMWW group were over-represented by the regulation of viral genome replication-related GO-Terms, whereas down regulated genes were mainly involved in cholesterol and lipid metabolism. CONCLUSIONS: Our study showed how an industrial waste product can be recycled as a feed additive with a positive relapse. OMWW dietary supplementation can be a nutritional strategy to improve chicken performance and health, prevent intestinal damage, enhance innate immunity and regulate cholesterol metabolism and fat deposition.

Gene co-expression networks in liver and muscle transcriptome reveal sex-specific gene expression in lambs fed with a mix of essential oils.

BACKGROUND: Essential oil (EO) dietary supplementation is a new strategy to improve animal health. EO compounds have antiparasitic, antimicrobial, antiviral, antimycotic, antioxidant and anti-inflammatory proprieties. Nutrigenomics investigations represent innovative approaches in understanding the relation between diet effect and gene expression related to the animal performance. Few nutrigenomics studies have used a high-throughput RNA-Sequencing (RNA-Seq) approach, despite great potential of RNA-Seq data in gene expression quantification and in co-expression network analyses. Our aim is to use the potential of RNA-Sequencing data in order to evaluate the effect of an EO supplementary diet on gene expression in both lamb liver and muscle. RESULTS: Using a treatment and sex interaction model, 13 and 4 differentially expressed genes were identified in liver and muscle respectively. Sex-specific differentially expressed (DE) genes were identified in both sexes. Using network based analysis, different clusters of co-expressed genes that were highly correlated to the diet were detected in males vs. females, in agreement with DE analysis. A total of five regulatory genes in liver tissue associated to EO diet were identified: DNAJB9, MANF, UFM1, CTNNLA1 and NFX1. Our study reveals a sex-dependent effect of EO diet in both tissues, and an influence on the expression of genes mainly involved in immune, inflammatory and stress pathway. CONCLUSION: Our analysis suggests a sex-dependent effect of the EO dietary supplementation on the expression profile of both liver and muscle tissues. We hypothesize that the presence of EOs could have beneficial effects on wellness of male lamb and further analyses are needed to understand the biological mechanisms behind the different effect of EO metabolites based on sex. Using lamb as a model for nutrigenomics studies, it could be interesting to investigate the effects of EO diets in other species and in humans.

Influence of Maternal and Postweaning Linseed Dietary Supplementation on Growth Rate, Lipid Profile, and Meat Quality Traits of Light Sarda Lambs.

The effects of dietary extruded linseed (EL) on growth performance, meat quality, and lipid profile of Semimembranosus and Longissimus lumborum muscles of 81 Sarda lambs were studied in a 3 × 3 design: EL content (0%, 10%, and 20%) of maternal dietary concentrate fed from 20 d to parturition to 60 d of lactation and EL content (0%, 10%, 20%) of lamb concentrate fed after weaning for 30 d. The basal diet was composed of alfalfa and meadow hay during pregnancy and alfalfa hay during lactation. At slaughter, carcass and meat quality were evaluated. Sensory quality of Semimembranosus from 0% and 20% EL lambs was assessed. Both maternal and postweaning diets affected growth performance, with higher body weights recorded with the 10% EL concentrate. Carcass and meat quality were not affected by diet. Saturated and monounsaturated FA decreased and n-3 polyunsaturated FA increased with increasing EL content in lamb diet. An increase in vaccenic and rumenic acid was associated with the EL content of the maternal diet. Both diets increased the n-6/n-3 FA ratio. No differences in acceptability were detected by consumers among groups. It is concluded that EL supplementation and early life nutrition can influence performance and FA metabolism in growing lambs.

Influence of manufacturing procedure on the compositional and sensory properties of n-3 fatty acid-enriched pecorino cheese.

Pecorino cheeses were produced using two different manufacturing procedures (either from raw milk or from milk previously subject to a thermisation at 65 °C for 10 s with the addition of starter culture) using the milk obtained from three groups of ewes fed three different concentrates: (1) control, (2) control enriched with 100 g/kg of extruded linseed, and (3) control enriched with 200 g/kg of extruded linseed. All concentrates were administered with alfalfa hay ad libitum. The extruded linseed-enriched diets increased the concentration of n-3 fatty acids, conjugated linoleic acid, and monounsaturated and polyunsaturated acids in raw milk and thermised milk cheese and decreased the concentration of saturated fatty acids. Changes in the manufacturing procedure did not influence the fatty acid composition of the cheeses but markedly influenced their sensory properties. In particular, heat-treating the milk and adding a starter reduced the differences in cheese odour, flavour and toughness induced by the diet when raw milk was used. Cheeses made with thermised milk and the addition of starter culture were more uniform and obtained a higher score in the preference test compared with the corresponding raw milk cheeses. An appropriate manufacturing process, therefore, can be a possible strategy to obtain cheeses with improved health-promoting properties and an unaltered acceptance level by consumers.

Olive Mill Waste-Water Extract Enriched in Hydroxytyrosol and Tyrosol Modulates Host-Pathogen Interaction in IPEC-J2 Cells.

The dietary supplementation of olive oil by-products, including olive mill waste-water (OMWW) in animal diets, is a novel application that allows for their re-utilization and recycling and could potentially decrease the use of antibiotics, antimicrobial resistance risk in livestock species, and the occurrence of intestinal diseases. Salmonella serovar typhimurium is one of the most widespread intestinal pathogens in the world, causing enterocolitis in pigs. The aim of this study was to investigate the effect of an OMWW extract enriched in polyphenols (hydroxytyrosol and tyrosol) in the immune response of an intestinal porcine epithelial cell line (IPEC-J2) following S. typhimurium infection. Cells were pre-treated with OMWW-extract polyphenols (OMWW-EP, 0.35 and 1.4 µg) for 24 h and then infected with S. typhimurium for 1 h. We evaluated bacterial invasiveness and assayed IPEC-J2 gene expression with RT-qPCR and cytokine release with an ELISA test. The obtained results showed that OMWW-EP (1.4 µg) significantly reduced S. typhimurium invasiveness; 0.35 µg decreased the IPEC-J2 gene expression of IL1B, MYD88, DEFB1 and DEFB4A, while 1.4 µg down-regulated IL1B and DEFB4A and increased TGFB1. The cytokine content was unchanged in infected cells. This is the first study demonstrating the in vitro immunomodulatory and antimicrobial activity of OMWW extracts enriched in polyphenols, suggesting a protective role of OMWW polyphenols on the pig intestine and their potential application as feed supplements in farm animals such as pigs.

Dietary supplementation with olive oil co-products rich in polyphenols: a novel nutraceutical approach in monogastric animal nutrition.

In recent years, the increased demand for agri-food products to feed livestock species has stimulated research to identify novel solutions for the valorization of natural waste, according to the modern concept of a circular economy. Numerous studies have shown the use of plant-derived and agro-industrial co-products that are sources of bioactive molecules for preparing animal feeds. Supplementation with co-products derived from the extraction of olive oil (i.e., olive pomace, olive mill wastewater, olive cake and olive leaf) in diet has been widely considered in recent decades, because these wastes are produced in high quantity and their re-use represents an innovative economic and environmental strategy. Olive oil co-products are characterized by various bioactive molecules such as polyphenols, carbohydrates, proteins, and lipids. Among them, polyphenols are the nutraceuticals most studied, showing to promote health effects in both humans and animals. Olive oil co-products and their phenolic extracts have shown many beneficial and promising effects when added to the diets of monogastric animals, by improving performance parameters and maintaining the oxidative status of meat and derived products. This review provides an update on the use of olive co-products in monogastric animal (swine, poultry and rabbit) diets and their effects on the productive performance, meat quality characteristics and gut health status.

Effect of heating, microbial fermentation, and enzymatic hydrolysis of soybean meal on growth performance, nutrient digestibility, and intestinal microbiota of weaned piglets.

The macromolecular proteins, anti-nutritional factors, and allergens contained in soybean meal (SBM) have a negative impact on the growth of weaned piglets. The objective of this study was to investigate the effects of heating, microbial fermentation, and enzymatically hydrolyzed SBM on the growth performance, nutrient digestibility, serum biochemistry, intestinal morphology, volatile fatty acids, and microbiota of weaned piglets. After the preparation of soaked SBM (SSBM), enzymatically hydrolyzed SBM (ESBM), and microbial fermented and enzymatically hydrolyzed SBM (MESBM), 72 weaned piglets were randomly allocated to three groups for a 21-d trial. In the three groups, 17% of conventional SBM in basal corn-soybean meal diet was replaced by an equivalent amount of SSBM (control group), ESBM, or MESBM. The results showed that the contents of glycinin, ß-conglycinin, trypsin inhibitor, and proteins above 20 kDa were significantly decreased in ESBM and MESBM, compared with SSBM, and the surface of ESBM and MESBM had more pores and fragmented structure. In the second week and throughout the entire experimental period, the diarrhea index was reduced (P < 0.01) in ESBM and MESBM in contrast with SSBM. Furthermore, the inclusion of ESBM and MESBM in the diet improved the apparent total tract digestibility of dry matter and crude protein (P < 0.05), and increased the abundances of the genera Lactobacillus and Clostridium_sensu_stricto_1, respectively. Metagenomic sequencing further identified that members of six species of Proteobacteria, four species of Clostridiales, and three species of Negativiautes were enriched in the colon of piglets fed MESBM, while two bacterial species, Lachnoclostridium and Lactobacillus_points, were enriched in the colon of piglets fed ESBM. In conclusion, replacing SSBM with ESBM or MESBM in the diet decreased the diarrhea index, which could be associated with improved nutrient digestibility and microbial composition.

With the development of pig industry, liquid feeding is becoming more widely used. Therefore, this study explores that liquid-state fermentation through enzymatic hydrolysis and microbial fermentation reduces the level of antigenic protein in soybean meal (SBM). In the present study, dietary supplementation with enzymatically hydrolyzed SBM (ESBM) or microbial fermented and enzymatically hydrolyzed SBM (MESBM) effectively decreased diarrhea index, enhanced nutrient digestibility, and improved the composition and stability of intestinal flora in weaning piglets. Our study not only contributes to the efficient utilization of SBM, but also provides new insights into its application in liquid feeding for livestock farming.

Nonpharmacological Treatment Strategies for the Management of Canine Chronic Inflammatory Enteropathy-A Narrative Review.

Chronic inflammatory enteropathy (CIE) refers to a heterogeneous group of idiopathic diseases of the dog characterised by persistent gastrointestinal (GI) clinical signs. If conventional dietary treatment alone would be unsuccessful, management of CIE is traditionally attained by the use of pharmaceuticals, such as antibiotics and immunosuppressive drugs. While being rather effective, however, these drugs are endowed with side effects, which may impact negatively on the animal's quality of life. Therefore, novel, safe and effective therapies for CIE are highly sought after. As gut microbiota imbalances are often associated with GI disorders, a compelling rationale exists for the use of nonpharmacological methods of microbial manipulation in CIE, such as faecal microbiota transplantation and administration of pre-, pro-, syn- and postbiotics. In addition to providing direct health benefits to the host via a gentle modulation of the intestinal microbiota composition and function, these treatments may also possess immunomodulatory and epithelial barrier-enhancing actions. Likewise, intestinal barrier integrity, along with mucosal inflammation, are deemed to be two chief therapeutic targets of mesenchymal stem cells and selected vegetable-derived bioactive compounds. Although pioneering studies have revealed encouraging findings regarding the use of novel treatment agents in CIE, a larger body of research is needed to address fully their mode of action, efficacy and safety.

Extracellular Vesicles from Animal Milk: Great Potentialities and Critical Issues.

Other than representing the main source of nutrition for newborn mammals, milk delivers a sophisticated signaling system from mother to child that promotes postnatal health. The bioactive components transferred through the milk intake are important for the development of the newborn immune system and include oligosaccharides, lactoferrin, lysozyme, α-La, and immunoglobulins. In the last 15 years, a pivotal role in this mother-to-child exchange has been attributed to extracellular vesicles (EVs). EVs are micro- and nanosized structures enclosed in a phospholipidic double-layer membrane that are produced by all cell types and released in the extracellular environment, reaching both close and distant cells. EVs mediate the intercellular cross-talk from the producing to the receiving cell through the transfer of molecules contained within them such as proteins, antigens, lipids, metabolites, RNAs, and DNA fragments. The complex cargo can induce a wide range of functional modulations in the recipient cell (i.e., anti-inflammatory, immunomodulating, angiogenetic, and pro-regenerative modulations) depending on the type of producing cells and the stimuli that these cells receive. EVs can be recovered from every biological fluid, including blood, urine, bronchoalveolar lavage fluid, saliva, bile, and milk, which is one of the most promising scalable vesicle sources. This review aimed to present the state-of-the-art of animal-milk-derived EV (mEV) studies due to the exponential growth of this field. A focus on the beneficial potentialities for human health and the issues of studying vesicles from milk, particularly for the analytical methodologies applied, is reported.

Blood thyroid hormones, insulin and leptin, metabolites and enzymes in transition dairy ewes, as affected by dietary linseed and physiological stage.

This study investigated the effects of dietary linseed and physiological state on blood concentrations of some metabolic hormones and indicators, in transition dairy ewes. From 21 d before lambing to 60 d post-partum, ewes were provided with one of three isoenergetic and iso­nitrogenous pelleted concentrates, without (CTR, n = 21), or with 100 (EL-10, n = 22) or 200 g/kg (EL-20, n = 22) of extruded linseed. Animals were fed alfalfa hay ad libitum and had access to mixed pasture. Plasma thyroid hormones and insulin concentrations were not affected by diet and significantly changed by time. The last week of pregnancy T3 (2.27 ± 0.15 ng/mL) and T4 (102.63 ± 2.85 ng/mL) were lower than at 4 weeks before lambing (T3: 3.27 ± 0.27 ng/mL; T4: 125.89 ± 2.63 ng/mL). After lambing, T3 rose to be highest at 4 and 8 weeks of lactation (3.41 ± 0.22). Mean insulin peaked 2 weeks after lambing (0.31 ± 0.02 ng/mL) vs. late pregnancy (0.22 ± 0.01 ng/mL) and progressing lactation (0.18 ± 0.01 ng/mL). Plasma Leptin concentration (2.43 ± 0.03 ng/mL) was not affected by diet nor time. All the blood metabolites and enzymes investigated showed significant time x treatment interaction. Differences of several haematological parameters were found in EL vs. CTR (cholesterol, triglycerides, total protein, bilirubin, ALP, ALT), however, in most cases the values were fairly within the reference physiological ranges. Metabolic hormones are confirmed to be mainly linked to the different physiological states, energy intake and variations of energy balance, without clear effects by different sources of energy and quality of dietary lipids.

Case Report: Oral Fecal Microbiota Transplantation in a Dog Suffering From Relapsing Chronic Diarrhea-Clinical Outcome and Follow-Up.

The present case report describes the effects of orally administered fecal microbiota transplantation (FMT) (frozen capsules) in a dog suffering from relapsing chronic diarrhea, needing a continuous low prednisolone dose to maintain the condition under acceptable control. Through FMT, we aimed at evaluating the possibility of improving the clinical score and/or reducing/suspending steroid administration. During a first period of strict monitoring (21 days), the canine inflammatory bowel disease activity index (CIBDAI) score passed from mild to clinically insignificant disease. Furthermore, two additional gastrointestinal signs that had been reported, bloating and episodes of painful defecation, rapidly improved (bloating) or even resolved (painful defecation). The patient was then followed for 18 months (to the authors' knowledge, the longest follow-up time ever reported in a dog), during which no serious relapses occurred and no increase in prednisolone dose was necessary. No adverse clinical effects were ever reported during monitoring. The present description provides a further experience increasing those already present in the veterinary literature, in which an agreement on how to use FMT has not yet been achieved although strongly needed and recommended.

Unveiling a hotspot of genetic diversity in southern Italy for the endangered Hermann's tortoise Testudo hermanni.

BACKGROUND: Hotspots of intraspecific genetic diversity represent invaluable resources for species to cope with environmental changes, and their identification is increasingly recognized as a major goal of conservation ecology research. However, even for iconic and endangered species, conservation strategies are often planned without thorough information on the geographic patterns of genetic variation. Here, we investigated the spatial patterns of genetic variation of the endangered Hermann's tortoise Testudo hermanni in the Italian Peninsula by genotyping 174 individuals at 7 microsatellite loci, with the aim to contribute to planning effective conservation strategies. RESULTS: Ordination-based and Bayesian clustering analyses consistently identified three main genetic clusters, one spread in the central and northern part of the peninsula, and two restricted to southern Italy and Sicily, respectively. The highest levels of genetic diversity were found in populations of the southern cluster and, in particular, at the northern edges of its distribution (He > 0.6, Ar > 2.8 ), that correspond to areas of putative secondary contact and admixture between distinct lineages. Our results clearly identify a hotspot of genetic diversity for the Hermann's tortoise in southern Italy. CONCLUSION: We inferred the evolutionary history and the spatial patterns of genetic variation of the Hermann's tortoise in the Italian Peninsula. We identified three main genetic clusters along the peninsula and a hotspot of intraspecific diversity in southern Italy. Our results underline the urgent need for conservation actions to warrant the long-term persistence of viable tortoise populations in this area. Furthrmore, these data add further evidence to the role of southern Italy as a biodiversity hotspot for temperate fauna, claiming for higher consideration of this area in large scale conservation programs.

Detection of Testudinid alphaherpesvirus, Chlamydia spp., Mycoplasma spp., and Salmonella spp. in free­ranging and rescued Italian Testudo hermanni hermanni.

Testudo hermanni is included as near­threatened in the Red List of the International Union for Conservation of Nature, while T. hermanni hermanni is considered endangered in the Italian Red List. Appropriate management of smuggled or seized wild individuals is recommended before their reintroduction into the wild. Accordingly, a health monitoring study was carried out. During 2014­2016, 133 oral swabs and 121 cloacal swabs were collected from a total of approximately 180 free­ranging and rescued T. hermanni hermanni from eight different Italian regions to investigate the presence of DNA of Testudinid alphaherpesvirus (TeAHV), Chlamydia spp. and Mycoplasma spp. in the oral cavity, and Salmonella spp. isolates in the cloaca. Mycoplasma spp. was detected in 52 out of 87 (59.77%) of rescued and in 1 out of 46 free­ranging (2.17%) individuals; 33 out of 53 (62.26%) Mycoplasma spp. positive samples were typed as M. agassizii by PCR. Salmonella spp. was isolated from 45 out of 121 (37.19%) cloacal swabs, typed into 14 serovars, and characterized for complete antimicrobial susceptibility. A significantly different distribution of Salmonella spp. isolates was found in 2016 in comparison with 2014 and 2015, without any difference between free­ranging and rescued tortoises. All the tested tortoises were negative for TeAHV and Chlamydia spp. These results are considered a baseline information critical to monitor the dynamics of these microorganisms in free­ranging and rescued populations of T. h. hermanni, and to correctly approach the management of rescued animals and possible relocation programs.

Dietary Supplementation of Olive Mill Waste Water Polyphenols in Rabbits: Evaluation of the Potential Effects on Hepatic Apoptosis, Inflammation and Metabolism through RT-qPCR Approach.

Agro-industrial processing for the production of food or non-food products generates a wide range of by-products and residues rich in bioactive compounds including polyphenols. The concentration of these by-products is sometimes higher than in the original raw material as in the case of olive mill waste water (OMWW), one of the main by-products of olive oil extraction. Polyphenols are secondary plant metabolites that regulate the expression of specific inflammatory genes, transcriptional factors and pro/anti-apoptotic molecules, thus modulating the signaling pathways essential for cell health and homeostasis. The liver plays a key role in regulating homeostasis by responding to dietary changes in order to maintain nutritional and physiological states. In this study a nutrigenomic approach was adopted, which focuses on the effects of diet-health-gene interactions and the modulation of cellular processes, in order to evaluate the expression of the genes (AGER, BAX, COX2, IL1B, PPARA, PPARG, SIRT1, TNFA) involved in these interactions in the livers of rabbits fed with a diet supplemented with OMWW (POL) or without supplements (control, CTR). The RT-qPCR analysis showed the down-regulation of SIRT1, TNFA, AGER, BAX and PPARA transcripts in the POL group compared to the CTR group. These results show that OMWW dietary supplementation prevents cell death and tissue deterioration in rabbits.

Differential Effects of Dietary Oregano Essential Oil on the Inflammation Related Gene Expression in Peripheral Blood Mononuclear Cells From Outdoor and Indoor Reared Pigs.

Intensive farming systems represent a stressful environment for pigs and negatively influence neuroendocrine functions, behavior, and performance. Outdoor farming is an alternative option, which is thought to imply several beneficial effects for the animal. Dietary essential oils are known to be an innovative strategy to improve pig health and performance, and oregano essential oil (ORE) possesses beneficial effects due to its antimicrobial, anti-fungal, and antioxidant properties. We tested the effect of dietary ORE on peripheral blood mononuclear cells (PBMCs) in 36 growing pigs, either reared under indoor or outdoor conditions. Quantitative real-time PCR (RT-qPCR) assay was used to evaluate the effect of diet (control vs. ORE) and the time of sampling (T1-120 days vs. T2-190 days) on the expression of inflammatory and immune-related genes (TNF, IL1ß, IL8, IL18, IL10, IL1RN, STAT3, HSP90, ICAM-1, and NFKB1). Under outdoor condition, the majority of transcripts were upregulated (p < 0.05), assuming a general inflammatory status (TNF, HSP90, NFKB1, IL1ß, and STAT3). However, an interaction between diet and the farming system was observed: HSP90, NFKB1, and STAT3 were downregulated (p < 0.05) in the outdoor reared pigs when fed the ORE diet. Our study showed that bioactive compounds of ORE exert their activity, especially when the animals are exposed to stressful stimuli. Dietary ORE can be an acceptable strategy to help pigs tolerate the stress related to the harsh, outdoor, rearing conditions.

The Use of Ascophyllum nodosum and Bacillus subtilis C-3102 in the Management of Canine Chronic Inflammatory Enteropathy: A Pilot Study.

The aim was to assess the effects of Ascophyllum nodosum (AN) with/without Bacillus subtilis C-3102 as alternative treatments for Chronic Inflammatory Enteropathy (CIE) of dogs. Fourteen CIE patients, which had received the same control (CTR) diet, were enrolled to serially receive three diets: (1) hydrolysed protein (HP) diet; (2) 4.0% AN supplemented HP (HPA) food, (3) HPA diet fortified with 125 billion B. subtilis C-3102 spores/10 kg body weight (HPAB diet). Clinical outcome was assessed by Canine Inflammatory Bowel Disease Activity Index (CIBDAI), whereas gut microbiota compositional variations were investigated via 16S rRNA gene analysis, and faecal fermentation end-products by liquid chromatography. Higher abundances of the Ruminococcaceae and Rikenellaceae families were shown in HPA relative to CTR treatment, with Bacillus genus being differentially abundant on HPAB diet. Concentrations of acetate were higher (p < 0.05) in dogs fed HPA compared to CTR diet, and amounts of isovalerate and isobutyrate were greater (p < 0.05) in HPA compared to HP food. A tendency for higher amounts of faecal butyrate was found for the HPAB treatment (p = 0.06). Comprehensively, while displaying potentially positive effects on faecal fermentations, the tested substances failed to improve CIBDAI scores and microbial richness in CIE dogs.

Mapping the geographic origin of captive and confiscated Hermann's tortoises: A genetic toolkit for conservation and forensic analyses.

The illegal trade has been threatening tortoise populations worldwide for decades. Nowadays, however, DNA typing and forensic genetic approaches allow us to investigate the geographic origin of confiscated animals and to relocate them into the wild, providing that suitable molecular tools and reference data are available. Here we assess the suitability of a small panel of microsatellite markers to investigate patterns of illegal translocations and to assist forensic genetic applications in the endangered Mediterranean land tortoise Testudo hermanni hermanni. Specific allelic ladders were created for each locus and tested on several reference samples. We used the microsatellite panel to (i) increase our understanding of the population genetic structure in wild populations with new data from previously unsampled geographic areas (overall 461 wild individuals from 28 sampling sites); (ii) detect the presence of non-native individuals in wild populations; and (iii) identify the most likely geographic area of origin of 458 confiscated individuals hosted in Italian seizure and recovery centers. Our analysis initially identified six major genetic clusters corresponding to different geographic macro-areas along the Mediterranean range. Long-distance migrants among wild populations, due to translocations, were found and removed from the reference database. Assignment tests allowed us to allocate approximately 70 % of confiscated individuals of unknown origin to one of the six Mediterranean macro-areas. Most of the assigned tortoises belonged to the genetic cluster corresponding to the area where the respective captivity center was located. However, we also found evidence of long-distance origins of confiscated individuals, especially in centers along the Adriatic coast and facing the Balkan regions, a well-known source of illegally traded individuals. Our results clearly show that the microsatellite panel and the reference dataset can play a beneficial role in reintroduction and repatriation projects when confiscated individuals need to be re-assigned to their respective macro-area of origin before release, and can assist future forensic genetic applications in detecting the illegal trade and possession of Testudo hermanni individuals.

Effects of Dietary Polyphenols from Olive Mill Waste Waters on Inflammatory and Apoptotic Effectors in Rabbit Ovary.

The aim of this study was to evaluate the effect of dietary polyphenols on the expression of the effectors involved in inflammation and apoptosis in rabbit ovary. New Zealand White female rabbits were fed a basal control diet (CTR), or the same diet supplemented with a polyphenolic concentrate (POL, 282.4 mg/kg) obtained from olive mill waste waters. The follicle counts and the relative mRNA (RT-qPCR) and protein (immunohistochemistry) expression of the effectors involved in inflammation (cyclooxygenase-2; interleukin-1beta; tumor necrosis factor-alpha, TNFA) and apoptosis (BCL2-associated X protein, BAX), detected in the ovaries of both groups, were examined. The POL diet increased the primary and total follicles number. Cyclooxygenase-2 gene expression was higher (p < 0.05) in the POL group than in the CTR group, whereas BAX was lower (p < 0.05) in POL than CTR. Immunohistochemistry revealed the presence of all the proteins examined, with weaker (p < 0.05) COX2 and BAX signals in POL. No differences between the CTR and POL groups were observed for IL1B and TNFA gene and protein expression. These preliminary findings show that dietary polyphenols modulate inflammatory and apoptotic activities in rabbit ovary, regulating cyclooxygenase-2 and BAX expression, thus suggesting a functional involvement of these dietary compounds in mammalian reproduction.

Plasmatic Profile of Pregnancy-Associated Glycoprotein (PAG) during Gestation and Postpartum in Sarda and Lacaune Sheep Determined with Two Radioimmunoassay Systems.

This study was carried out to determine ovine pregnancy-associated glycoprotein (oPAG) levels in the plasma of Sarda and Lacaune ewes throughout gestation and in the first month postpartum, using two heterologous radioimmunoassays (RIA-706 and RIA-srPool) and to study the correlations between PAG levels and fetal gender and number. On Day 18 of pregnancy, PAG concentrations were detected in 90.1% and 80.8% of Sarda pregnant ewes with RIA-706 and RIA-srPool, respectively; and in 90% and in 75% of Lacaune pregnant ewes with RIA-706 and RIA-srPool, respectively. From Day 30, PAG concentrations were detected in all pregnant ewes by using both RIA methods. In the postpartum period, the PAG concentrations in Sarda ewes decreased rapidly reaching minimal levels (<1 ng/mL) on day 28 using both RIA-706 and RIA-srPool. In Lacaune ewes, PAG-706 levels were higher than PAG-srPool from parturition until the last day of observation (Day 28 postpartum). It was also observed that mean concentrations were higher in multiple than in single pregnancies in Sarda and Lacaune ewes. Moreover, due to the rapid disappearance of PAG concentration following lambing, a cut-off limit in postpartum animals was not required as a means for detecting a new pregnancy.