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Background: Nerve Growth Factor (NGF) plays an important role in the reproductive system through its receptor's interaction (p75 NTR ). This paper aims to analyze the impact of NGF p75 NTR in epididymal and ejaculated rabbit semen during in vitro sperm storage., Methods: Semen samples from 10 adult rabbit bucks were collected four times (n = 40) and analyzed. NGF was quantified in seminal plasma, and the basal expression of p75 NTR in sperm was established (time 0). Moreover, we evaluated p75 NTR , the apoptotic rates, and the main sperm parameters, at times 2-4 and 6 h with or without the administration of exogenous NGF., Results: Based on the level of p75 NTR , we defined the threshold value (25.6%), and sperm were divided into High (H) and Normal (N). During sperm storage, p75 NTR of H samples significantly modulated some relevant sperm parameters. Specifically, comparing H samples with N ones, we observed a reduction in motility and non-capacitated cell number, together with an increased percentage of dead and apoptotic cells. Notably, the N group showed a reduction in dead and apoptotic cells after NGF treatment. Conversely, the NGF administration on H sperm did not change either the percentage of dead cells or the apoptotic rate., Conclusion: The concentration of p75 NTR on ejaculated sperm modulates many semen outcomes (motility, apoptosis, viability) through NGF interaction affecting the senescence of sperm.

The metabolism of polyunsaturated fatty acids (PUFAs) plays an important role in male reproduction. Linoleic and alpha-linolenic acids need to be provided in the diet and they are converted into long chain polyunsaturated fatty acids by steps of elongation and desaturation, exerted by elongases 2 (ELOVL2) and 5 (ELOVL5) and Δ5- (FADS1) and Δ6-desaturase (FADS2). This study aims to assess the gene expression and localization of enzymes involved in the synthesis of n-3 and n-6 long-chain PUFAs in control rabbits and those fed diets containing 10% extruded flaxseed. Enzyme and PUFA localization were assessed in the testes and epididymis by immunofluorescence. Testes showed high gene expression of FADS2, ELOVL2 and ELOVL5 and low expression of FADS1. Intermediate metabolites, enzymes and final products were differently found in Leydig, Sertoli and germinal cells. FADS2 was localized in interstitial cells and elongated spermatids; ELOVL5 in meiotic cells; FADS1 was evident in interstitial tissue, Sertoli cells and elongated spermatids; ELOVL2 in interstitial cells. Epididymal vesicles were positive for FADS1, ELOVL2 and ELOVL5 as well as docosahexaenoic, eicosapentaenoic, and arachidonic acids. This knowledge of fatty acids (FA) metabolism in spermatogenesis and the influence of diet on FA profile could help identify causes of male infertility, suggesting new personalized therapy., (© 2022. The Author(s).)

The present study evaluated the effects of feed supplemented with two dietary sources of n-3 polyunsaturated fatty acids (PUFAs; fish oil and extruded flaxseed) on the gut microbiota, caecal fermentations, gastrointestinal histology, and histochemistry in rabbits. Fifteen male New Zealand White rabbits were divided into three groups ( n = 5/group) and fed with different diets from weaning (35 days of age) until slaughtering (90 days of age): C group, fed with a commercial diet; F group, supplemented with 10% of extruded flaxseed; and O group, supplemented with 3.5% of fish oil. At slaughter, the content of the stomach, duodenum, jejunum, ileum, caecum, and colon was collected and analyzed by Next Generation 16S rRNA gene sequencing. Tissue samples of the same tracts were evaluated with histological and histochemical analysis. Ammonia and lactic acid in the caecum were also quantified. Twenty-nine operational taxonomic units (OTUs) were significantly different between groups. Groups receiving n-3 PUFAs supplementation showed an increase in Bacteroidetes and Lachnospiraceae in several gastrointestinal tracts, while Bacilli abundance, as well as Firmicutes/Bacteroidetes ratio, were reduced compared to the control group (for all p < 0.05). Caecal ammonia was lower in the F than C group ( p < 0.032), whereas no difference was found for lactic acid. Finally, histological evaluations revealed a mild hemorrhagic infiltration and vessels ectasia in the stomach mucosa of both F and O groups, but no effect of nutritional treatment was evidenced by the histochemical analyses. In conclusion, n-3 PUFAs supplementation could modify the rabbit gut microbiota and fermentation. The increase in beneficial bacterial populations may, at least partially, explain the positive effects of n-3 PUFAs diet supplementation on human and animals' health, although the appropriate dosage should be established.

In the last decades, several nutraceutical substances have received great attention for their potential role in the prevention and treatment of different diseases as well as for their beneficial effects in promoting the health of humans and animals. Goji berries (GBs) are the fruit of Lycium barbarum and other species of Lycium , used in traditional Chinese medicine, and they have recently become very popular in the Occidental world because of their properties, such as anti-aging, antioxidant, anticancer, neuroprotective, cytoprotective, antidiabetic, and anti-inflammatory activities. These effects are essentially evaluated in clinical trials in humans; in experimental animal models, such as mice and rats; and in cell lines in in vitro studies. Only recently has scientific research evaluated the effects of GBs diet supplementation in livestock animals, including rabbits. Although studies in the zootechnical field are still limited and the investigation of the GB mechanisms of action is in an early stage, the results are encouraging. This review includes a survey of the experimental trials that evaluated the effects of the GBs supplementation on reproductive and productive performances, immune system, metabolic homeostasis, and meat quality principally in the rabbit with also some references to other livestock animal species. Evidence supports the idea that GB supplementation could be used in rabbit breeding, although future studies should be conducted to establish the optimal dose to be administered and to assess the sustainability of the use of GBs in the diet of the rabbit., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Agradi, Draghi, Cotozzolo, Barbato, Castrica, Quattrone, Sulce, Vigo, Menchetti, Ceccarini, Andoni, Riva, Marongiu, Curone and Brecchia.)

Goji berries show health benefits, although the possible mechanisms of action, including compositional changes in the gut microbiome, are still not fully understood. The aim of this study was to evaluate the effect of Goji berry supplementation on microbiota composition and metabolites in the digestive tracts of rabbits. Twenty-eight New Zealand White rabbits were fed with a commercial feed (control group, C; n = 14) or the same diet supplemented with 3% of Goji berries (Goji group, G; n = 14), from weaning (35 days old) until slaughter (90 days old). At slaughter, samples from the content of the gastrointestinal tracts were collected and analyzed by Next Generation 16S rRNA Gene Sequencing to evaluate the microbial composition. Ammonia and lactic acid were also quantified in caecum. Results showed differences in microbiota composition between the groups for two phyla (Cyanobacteria and Euryarchaeota), two classes (Methanobacteria and Bacilli), five orders, fourteen families, and forty-five genera. Ruminococcaceae ( p < 0.05) and Lachnospiraceae ( p < 0.01) were more abundant in G than in C group. Lactobacillaceae also showed differences between the two groups, with Lactobacillus as the predominant genus ( p = 0.002). Finally, Goji berry supplementation stimulated lactic acid fermentation ( p < 0.05). Thus, Goji berry supplementation could modulate gastrointestinal microbiota composition and caecal fermentation.

The aim of this study was to analyze the relationship between the kinetic behavior, carcass characteristics, oxidative status (blood and meat), and meat fatty acid profiles of 6 organically reared slower growing chicken genotypes (SrG). One hundred male chickens of 6 SrG were used: Ranger Classic (RC), Ranger Gold (RG), Rowan Ranger (RR), RedJA (RJ), CY Gen 5 JA87 (CY), and M22 × JA87 (M). Twenty chickens/genotype were selected to analyze behavior, while, 15 individuals were slaughtered and different traits were analyzed in the blood and drumstick meat. The variables were grouped into different principal components: kinetic activity (PC1, with explorative attitude as the highest score), productive performance (PC2, carcass and head/feet yields), blood (PC3, carbonyls, and TBARS) and meat (PC6, thiols, and TBARS) markers, technological traits (PC4, pH, and color), proximate meat composition (PCA5, moisture, lipids, protein, and ash), fatty acid profile, and nutritional indexes (PC7, IP, and PUFAn-3). Uni- and bivariate analyses showed a strong positive association between kinetic behaviors and blood and meat oxidation and a medium positive association with fatty acid profile and nutritional indexes, whereas a negative association was found between productive performance loads and the technological traits of meat. Generalized linear models showed that all PCs were influenced by genotype. In particular, CY and M resulted as less active genotypes; conversely, RR showed more kinetic activity, whereas RJ, RG, and RC exhibited intermediate levels of activity. Cluster analysis of kinetic behavior and blood or meat oxidative status highlighted 2 groups: nonwalking (NW: CY and M) and walking (W: RC, RG, RR, and RJ) animals. However, in the W group, another was visualized, constituted by genotypes with high kinetic activity resulting in the worst oxidative balance (Walking not trained-genotypes, Wnt: RR and RJ). The present results confirmed that the kinetic behavior of SrG genotypes is negatively correlated with productive performance. Furthermore, a significant association between kinetic behavior and blood (positively correlated) or meat (negatively correlated) oxidative status was noted. Such differences are mainly due to the intrinsic response of the genotypes used (i.e., training-walking capacity)., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Goji berry shows a wide range of beneficial properties in human health, but only a few studies evaluated its effects in livestock animals. The objective of this research was to assess the effects of goji berry supplementation on the hormonal profile, productive, and reproductive performance of does. Two months before artificial insemination, 105 nulliparous does were randomly divided into three groups ( n = 35) based on the dietary treatment: commercial diet (C), or a diet supplemented with either 1% (G1), or 3% (G3) of goji berry, respectively. The results showed that receptivity was higher in G1 than in the C group ( p < 0.05). Trends toward significance for differences between the G1 and G3 groups in marginal means of LH concentrations ( p = 0.059), and between G1 and C in LH AUC values ( p = 0.078), were evidenced. Estrogen concentrations showed a more fluctuating trend but a significant interaction effect ( p < 0.001). The G1 group showed higher litter weight than C at birth ( p = 0.008) and weaning ( p < 0.001), as well as higher litter size at weaning ( p = 0.020). The G1 group also exhibited the highest mean milk production ( p < 0.01). In conclusion, goji berry influenced reproductive and productive performance, probably via modulating hormonal patterns and milk production in rabbits. However, further studies are needed to validate these preliminary results.

The demand for poultry meat, being cheaper than red meat, will drive worldwide production of this product. Accordingly, an increase in production up to 16% is expected in 2025, most of which will occur in developing countries. Most poultry meat production is realized with intensive production systems, and extensive rearing systems (ERS) of poultry (organic, free-range, and low-input) represent only a small portion of poultry production in the EU (about 5%). However, there is an increasing interest in such rearing systems to maintain the good image of product and environmental sustainability, improved animal welfare, and meat quality with an annual trend of growth of about 10%. The aims of this work were to summarize the activities and the viewpoint of the researchers of the Department of Agricultural, Food, and Environmental Science of the University of Perugia (Italy). One of the most important goals of the research unit was the challenge of identifying the best poultry genotypes for ERS, which are important not only for the food industry but also for the improvement of human nutrition. Only the definition of the best genotypes adapted to ERS through the measurement of a wide panel of traits-genetic, physiologic, and behavior-and not only relying on daily weight gain will allow us to achieve this goal., Competing Interests: The authors declare no conflict of interest.

Polyunsaturated fatty acid (PUFA) metabolism and tissue distribution is modulated by the oxidation of these molecules. This research aimed to investigate the implication of dietary n-3 PUFA supplementation (precursor and long-chain PUFA) on the PUFA profile and oxidative status of the liver, testis, and brain of adult rabbit bucks. Twenty New Zealand White rabbit bucks were divided into four experimental groups ( n = 5 per group) and were fed different diets for 110 days: control (CNT), standard diet containing 50 mg/kg alpha-tocopheryl acetate (vitamin E); CNT+, standard diet + 200 mg/kg vitamin E; FLAX, standard diet + 10% flaxseed + 200 mg/kg vitamin E; or FISH, standard diet + 3.5% fish oil + 200 mg/kg vitamin E. Antioxidants (enzymatic and non-enzymatic), oxidative status (malondialdehyde and isoprostanoids), and n-3 and n-6 PUFAs of tissues were analysed. A chain mechanism of oxidant/antioxidant molecules, which largely depended on the particular PUFA composition, was delineated in the different organs. The liver showed an oxidant/antioxidant profile and lipid pathways widely modulated by PUFA and vitamin E administration; on the other hand, the testis' oxidative profile rather than its lipid profile seemed to be particularly affected, an outcome opposite to that of the brain (modulation operated by dietary PUFA).

Forty-two New Zealand White rabbits ( n = 21/group) were fed with two different diets: a commercial diet (control group) and a diet supplemented with goji berries (3% w / w ). After slaughtering, the effect of dietary supplementation on microbiological, physico-chemical, and sensory characteristics of the rabbit loins, packed in an oxygen-permeable package, was evaluated at 6 h post mortem (day 0), after 4 and 10 days of refrigerated storage. No relevant results were obtained for pH and total volatile basic Nitrogen (TVBN) values but with regards to the color, some significant differences were observed between the groups. The goji berries (GBs) dietary supplementation had positive effects by reducing thiobarbituric acid reactive substances (TBARS) values in all the observations ( p < 0.001). Moreover, microbiological results showed that the supplementation had a significant impact on Lactobacillus spp. ( p < 0.001) prevalence, indeed the goji group had higher means on day 0 ( p < 0.05) and on day 4 ( p < 0.001) than the control group. Lastly, with regards to the consumer's test, the tasters assigned a higher score to GBs rabbit meatballs and the purchase interest increased when the rabbit diet was known. Overall, these results indicate that the goji berries inclusion in the rabbit diet could represent a valuable strategy to improve quality and sensory traits of meat.

The influence of NGF in male reproduction in some animal species and humans has already been assessed. Many of these effects are mediated by the distribution and abundance of tropomyosin receptor kinase A (TrKA) and p75 neurotrophin (p75NTR) receptors on sperm cells. The aim of this research was to investigate the role of NGF and its receptors, TrKA and p75NTR, in rabbit sperm outcomes during in vitro storage. Major semen traits (kinetic parameters, apoptotic, necrotic and live sperm) were recorded in rabbit semen samples from 0 to 12 h of storage (every 4 h). Three experimental hypotheses were formulated: i) sperm storage changes NGF receptor abundance in rabbit sperm; ii) TrKA and p75NTR differently modulate NGF signalling (assessed by the neutralisation of receptors); iii) NGF-receptor interactions show different responses during storage (evaluated by the addition of exogenous NGF). The results demonstrate that: (i) the receptor number changed in a time-dependent manner with a significant increase in p75NTR after 8-12 h of storage; ii) the neutralisation of NGF receptors largely affected VCL, apoptotic, necrotic and live cells during sperm storage, i.e. blockade of TrKA significantly increased speed, capacitation, necrosis and apoptosis, whereas blockade of p75NTR improved motility and live cells; iii) the addition of exogenous human NGF (100 ng/mL) at different time points of storage (0, 4, 8 h) differently influenced sperm traits i.e. NGF addition at time 0 positively affected all the pro-vital traits (kinetic, live cells) whereas, after 4-8 h, the effect of NGF was null or negative. In conclusion, NGF affects kinetic and other physiological traits (capacitation, apoptosis and necrosis) of rabbit sperm in a time-dependent manner. Most of these modifications are modulated by the receptors involved (TrKA or p75NTR), which changed considerably during sperm storage (increase of p75NTR)., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The objective of this study was to compare the effect of the dietary beer trub, alone or in combination with linseed, on meat quality, oxidative status and cholesterol content of rabbit. Eighty New Zealand White rabbits were divided at weaning (30 d) into four dietary groups: control (C) fed a standard diet, trub (T, 2% of lyophilized trub), linseed (L, 3% of extruded linseed), and trub-linseed in combination (TL, 2% of lyophilized trub and 3% of extruded linseed). At slaughtering (80 d) the oxidative parameters of blood and quality of Longissimus thoracis et lumborum were analysed. The in vivo lipid oxidation was similar in the supplemental groups, whereas it was higher in T, L and TL meat compared to C. Trub supplementation in rabbit diets slightly affected the fatty acid profile (MUFA) and worsened the oxidative status of meat, and its inclusion in combination with linseed did not counteract the oxidative instability induced by the higher PUFA content., Competing Interests: Declaration of Competing Interest We declare none conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The aim of this study was to verify the motivation of rabbit does to social contact or seclusion. The results of two different research activities assessed in Italy (experiment 1) and Germany (experiment 2) through the use of motivational cages are reported. In experiment 1, only the average time of occupation of the group or seclusion zone was recorded of four nulliparous does, while, in experiment 2, the group-housing system provided space for does with kits and consisted of four single areas (nest boxes with individual electronic nest box recognition systems). Experiment 1 showed that does spent a similar amount of time in seclusion or in group (49.61% vs 50.39%, respectively). On the contrary, in experiment 2, does with kits appeared to prefer spending time alone (71.90%) rather than in groups. The presence of kits probably stimulates a hierarchical and aggressive response of the dominant does, with the low-ranking does staying secluded to avoid violent interactions. In fact, in each reproductive cycle, one doe did stay in the group area whereas the other three does used this area in different percentages of time. Further researches are needed to find a good combination of the cage with the does' physiological phases., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

In the last two decades, the human sperm count linearly decreased in Western countries. Health problems, lifestyle, pollutants, and dietary behaviours are considered as the main risk factors, and the unbalance of dietary n -6/ n -3 fatty acids is one of the most relevant. The aim of the present research is to study the effect of different dietary sources of n -3 polyunsaturated fatty acids (PUFA) on reproductive traits using rabbit buck as the animal model. Fifteen rabbit bucks were assigned to three experimental groups: the control group, the FLAX group fed 10% extruded flaxseed, and the FISH group fed 3.5% fish oil for 110 days (50-day adaptation and 60-day experimental periods). Semen samples were collected weekly, whereas blood was collected every two weeks for the analytical determination of semen traits, oxidative status, fatty acid profiles, isoprostanes, neuroprostanes, and the immunocytochemistry of docosahexaenoic acid (DHA) and eicosapentaenoic (EPA) acid. At the end of the trial, the rabbits were killed and the testes were removed and stored for the analysis of fatty acid profile and immunocytochemistry. Results showed that dietary administration of n -3 PUFA improved the track speed of the sperm and increased the n -3 long-chain PUFA mainly confined in the sperm tail. Seminal plasma increased the thiobarbituric reactive substances (TBARs) by three times in the groups fed supplemental n -3, whereas the F 2 -isoprotanes (F 2 -IsoPs) and F 4 -neuroprostanes (F 4 -NeuroPs) were lower and higher, respectively, in both supplemented groups than in the control. The testes and sperm showed a higher DHA and EPA distribution in rabbits from the n -3 supplemented groups compared with the control. In conclusion, supplemental dietary n -3 PUFA improved sperm motion traits and resulted in an enrichment of membrane fatty acid in the sperm and testes of the rabbits. However, such an increased amount of PUFA negatively affected the sperm oxidative status, which was mainly correlated with the generation of F 4 -NeuroPs with respect to F 2 -IsoPs. Accordingly, the latter cannot be considered a good marker of oxidation when diets rich in n -3 PUFA are provided., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2019 Cesare Castellini et al.)

The improvement of rabbit gut microbiota by modifying nutritional components of the feed or favoring its early intake of feed has been previously investigated. The early administration of dehydrated alfalfa (A) or fresh grass (G) for rabbits, during the peri-weaning period (30 and 45 days of age), and their effect on performance and caecal microbiota compared to a standard diet (C) were evaluated. Until 15 days of age, nine litters/group were housed in the maternal cage and milked once per day. From 15 to 30 days, the young rabbits could consume both milk and solid feed (pelleted for C or supplemental feed for A and G). At 30 days of age, the rabbits were weaned and, until 45 days, were kept in single cages following the same dietary protocol. No significant changes were found in the milk intake or the individual weight of young rabbits at 30 and 45 days. The caecal Firmicutes/Bacteroidetes (bacterial phyla ratio) increased with age (from 2.43 to 6.05 on average, at 30 and 45 days). The Ruminococcaceae/Lachinospiraceae (bacterial family ratio) was highest in the A group at both ages, followed by G then C. The early administration of dehydrated alfalfa is a promising solution to improve health status by favoring an appropriate digestive microbiota.

Background: Fresh sprouts are healthy foods, low in fats and high in phytochemicals, but have a short shelf-life, hence the need for processing methods that preserve their nutritional value. This work was aimed at evaluating the effect of heat-drying (HD) and freeze-drying (FD) on the phytochemical and fatty acid profiles of alfalfa and flax sprouts, as compared to fresh material., Results: Both FD and HD reduced the phytochemical contents compared to fresh sprouts. FD better preserved phytoestrogens, phytosterols and total tocols compared to HD. However, phytoestrogen and tocol content remained quite high also in HD. The fatty acid profile was affected only by sprouts species, with higher amounts of α-linolenic acid in flax and linoleic acid in alfalfa sprouts., Conclusions: This work demonstrates that drying does not severely compromise the nutritional value of sprouts and provides a valid support for the choice of the drying method depending on the compound to be preserved, and taking into consideration the different cost of the methods. In addition, sprout powder is easy to handle and, due to its low volume, does not imply a decrease of feed consumption and energy intake, which is relevant in its use as a supplement in human and animal feeding. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Sixty New Zealand White weaned rabbits were divided into three groups and subjected to different dietary treatments: a standard diet for the control (C), a standard feed supplemented with 10% of plain olive leaves (OL) and a standard feed supplemented with 10% of selenium-fortified olive leaves (100 mg/L of foliar spray sodium selenate solution; SeOL). The productive performance was recorded at the time of slaughter (after 35 days); the carcass and meat traits were determined and estimated indexes of fatty acid metabolism were calculated. No significant differences were found on the rabbit productive performance and the physical-chemical characteristics of the meat. Both group of rabbits on the enriched diet showed leaner and thinner carcasses and a higher meat concentration of oleic acid. The estimated index of Δ5 + Δ6-desaturase, starting from n-6 fatty acids, was lower in both groups supplemented with leaves. The use of selenium-fortified olive leaves, positively affected the lipid oxidative stability of rabbit meat., (Copyright © 2018 Elsevier Ltd. All rights reserved.)