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This study provides a comparative assessment of fresh semen quality and sensitivity to cryopreservation in several Italian chicken breeds. The research involved 145 roosters from 13 breeds. The results showed a wide variability in all the parameters considered among the different breeds, especially in the quantitative variables of fresh semen, such as volume and concentration. For the qualitative characteristics (sperm membrane integrity and motility parameters), the variability across breeds was more pronounced for frozen than fresh semen. Interestingly, apart from total motility in fresh semen, breed had a significant effect on all semen quality parameters in both fresh and thawed ejaculates. Considering the overall qualitative characteristics, the Robusta maculata, Siciliana, and Mericanel della Brianza breeds produced ejaculates with better semen quality compared to other Italian breeds. By evaluating the main parameters of semen quality, our results underline the potential of these traits to influence the reproductive success and genetic conservation. The Bionda piemontese, Bianca di Saluzzo, Livorno bianca, Pepoi, and Siciliana breeds showed better resilience to cryopreservation, suggesting the need for breed-specific protocols to optimise semen quality after thawing. Importantly, the research highlights the central role of semen quality for both immediate fertilisation success and long-term conservation efforts. Future studies integrating OMICS technologies could elucidate molecular markers influencing breed-specific differences, helping to refine cryopreservation techniques and improve conservation strategies for indigenous Italian chicken breeds. This work contributes valuable insights to global efforts aimed at safeguarding poultry genetic diversity and sustainability.

Specific characteristics of the eggs from nucleus populations of Ancona (AN), Bianca di Saluzzo (BS), Bionda Piemontese (BP), Livorno, Mericanel della Brianza (MB), Mugellese (MU), Siciliana (SI) and Valdarnese (VA) chicken breeds kept in University Research Centres across Italy were evaluated. The physical characteristics considered were whole egg weight and shell, yolk and albumen proportion. The chemical characteristics considered were dry matter (DM), ash and protein content of the yolk and albumen, and the yolk fat. Among the breed groups evaluated in this study, BS and BP produce large eggs that are rich in total crude protein and lipids, with abundant egg whites. MB and MU produce small eggs with a high yolk percentage. AN and SI hens produce medium-small-sized eggs. The heaviest yolks were observed in AN eggs, and the highest shell percentage was found in SI eggs. LB and VA produce medium-large-sized eggs with low yolk lipid content.

The creation of genetic resource cryobanks provides a crucial link between in situ and ex situ techniques to improve the efficiency of conservation programs. Aim of the present review is to describe all the activities developed for the implementation of the first Italian Semen Cryobank of Autochthonous Chicken and Turkey Breeds. These activities can be classified into three main topics: (1) identification of species-specific semen freezing/thawing reference procedures; (2) drafting Standard Operative Procedures (SOP) for the implementation of the semen cryobank; (3) storage of semen doses from Italian chicken and turkey breeds to establish the cryobank. Several trials have been developed to identify a specie-specific semen cryopreservation protocol for chickens and turkeys. The major results are reviewed and a final reference protocol described. Taking into consideration the FAO guidelines for cryoconservation of animal genetic resources, SOP were drafted with the aim to provide technical guidance and logistical support on the choice of priority breeds, selection of birds for semen production, infrastructures and storage sites, birds and semen management, cryopreservation process and doses traceability. Lastly, the Italian Semen Cryobank was created. A total of 112 semen doses from 22 cockerels of three breeds, and 74 doses from 12 turkey males of three breeds were stored in the Cryobank. Breed specific semen quality parameters assessed before and after cryopreservation are reported. The described activities provide information and tools useful for the implementation of semen cryobanking in avian species and might be transferred also to other species after appropriate adaptations.HIGHLIGHTS Implementation of the first Italian Semen Cryobank of Autochthonous Chicken and Turkey Breeds Drafting Standard Operative Procedures provides technical guidance and logistical support on the design and establishment of the cryobank Semen cryobank is a precious genetic reservoir and could be useful to safeguard genetic variability in small population in vivo conserved

Maintenance or improvement of sperm quality during storage could prevent the loss of fertilizing capacity associated with stored turkey semen. Therefore the optimization of stored turkey semen could be useful to breeder industry since the commercial production of this bird relies almost entirely on artificial insemination. Previous research have shown that He-Ne laser irradiation in mammalian sperm increased the motility (Stato, 1986), decreased the mortality, promoted the acrosome reaction, which have a pivotal role in assisted fecundating programmes as therapy for resolving infertility in domestic animals..........

Considering the beneficial effects of n-3 polyunsaturated fatty acids (PUFA) for human health (Leskanich and Noble, 1997) several studies have been carried out to enrich animal products with these compounds. Both eggs and poultry meat enriched with n-3 PUFA may be considered as valid n-3 PUFA alternative sources to natural occurring fish products (Meluzzi et al., 2001; Sirri et al., 2002)...

This study was designed to compare Cortalap® extender with tris-citric acid-glucose (TCG), Lepus® and Merk III®, to assess the in vitro preservability of rabbit spermatozoa stored for 72 h at 5°C. The best extender identified in vitro was then compared with fresh semen to determine fertility and prolificacy rates in vivo. Eight semen pools were split into four subsamples and each of them was diluted to a ratio of 1:10 (volume:volume) with four different extenders (Lepus®, Cortalap®, TCG and Merk III®) and stored at 5°C for 72 h. Sperm motility, viability, sperm membrane and acrosome integrity were evaluated in both fresh and chilled semen at 4, 24, 48 and 72 h of storage. The results showed that Cortalap® was the best extender to preserve the quality of rabbit semen in vitro over 72 h in comparison with the other extenders (P® was used for an artificial insemination (AI) trial to be compared with fresh semen. Two groups of does (n=30 each) were inseminated with fresh and chilled semen. The fertility and prolificacy rates for the does inseminated with chilled semen were significantly lower compared with those inseminated with fresh semen (P® proved to be the best extender to preserve semen quality in vitro after 72 h at 5°C, the in vivo results showed that its use is not recommended in AI programmes.

Semen of the domestic turkey cannot be stored longer than 6 h without a loss of fertilizing capability. The improvement of long-term liquid storage procedures of semen is important since the commercial production of turkey relies almost entirely on artificial insemination. Therefore, studies improving storage regimens would allow longer storage and consequently hen fertility (Iaffaldano and Meluzzi, 2003). Since the search for an optimal extender composition for semen storage is still in progress, the aim of this paper has been to study the effects of different extenders on the quality of turkey semen during the storage for 48 h at 5°C.

The effects of different prefreezing semen concentrations on post-thawing quality of turkey semen cryopreserved with the pellet method were investigated. Ten pooled semen samples were each divided into 6 subsamples and diluted with Tselutin extender to obtain a final concentration of 0.5, 1, 2, 3, 4 and 5x109/mL respectively. Subsamples were cooled, added with 8% of dimethylaceta- mide as cryoprotectant and, after 5 minutes of equilibration, seminal aliquots of 80 μL were directly dropped into a liquid nitrogen bath to form frozen pellets. Thawing was performed in few seconds at 75°C. Sperm motility (Accudenz® swim-down test), viability (SyBr-Propidium Iodide staining) and sperm susceptibility to osmotic stress (Hyposmotic-water test) were assessed. Cryopreservation caused an overall loss of sperm quality, however differences in seminal parameters due to the different sperm concentration were observed in turkey spermatozoa after thawing: spermatozoa diluted to 4x109/mL showed significant higher values in mobility, viable and osmotic resistant spermatozoa compared to the other concentrations. This study showed that the post-thaw quality of turkey semen cryopreserved by pellets method was affected differently in relation to prefreezing sperm concentration.

Fish production requires great efforts and energies and causes different impacts/effects on the ecosystems. The development of aquaculture is possible, in the medium and long term, only if this is done responsibly, and if the production is reached from both an economic and environmental point of view. In our study we have worked on fry production using 1.5 g sized Dicentrarchus labrax obtained rearing larvae for 120 days after the hatching of the eggs.

Neuroglia has been historically considered the “glue” of the nervous system, as the ancient Greek name suggests, being simply referred as non-neuronal cells, with supporting functions for neurons in the CNS of mammalian and lower vertebrates. All around the world, approximately 283 cell lines were obtained from fish, yet none of these was from the brain of Sparus aurata, neither in cell lines nor as primary culture. Here we describe a novel in vitro reproducible neuroglial marine model for establishing primary neuroglial cell cultures, by dissociating the whole brain of seabream juveniles. We showed that proliferating neural stem cells produced alongside three generating lineages, such as neuronal precursor cells, astroglial precursor cells and oligodendroglia precursor cells, which developed respectively neurons, astrocytes and oligodendrocytes. The radial glia, finely described by morphological studies and immunochemical antigen expression, showed a peculiar spatial distribution, giving rise simultaneously both to astrocytes and neuronal precursors within a highly proliferative assemblate. Radial glia cells were assessed by glial fibrillary acidic protein (GFAP) and vimentin reactivity, astrocytes by GFAP, neurons by the neuron-specific markers for ubiquitin carboxy-terminal hydrolase 1 (UCHL1) and intermediate filament associated protein (NF), whereas myelinating oligodendrocytes were immunostained with anti-myelin basic protein (MBP) and anti-O4. Our findings suggest that seabream neuroglial cells gain in 3-4 weeks of culturing proliferation, neuroglial differentiation, and oligodendrocyte maturation with myelination, thus disclosing on the possibility that mixed neuroglial cultures can accelerate the maturation of oligodendrocytes and the regeneration of CNS injury in fish.

Among all know mycotoxins, aflatoxin B1 is one of the most studied for its hepatotoxic, carcinogenic, mutagenic, teratogenic, and immunosuppressant effects. This work aimed to study the effects of AFB1 acute and chronic exposure on CYP1A and GST enzymes induced in vitro on S. aurata hepatocytes by immunoblot analysis, thus relating the cytotoxic effects leading to cell death by apoptotic studies. Immunofluorescence analysis revealed that cell damage was not recoverable but permanent, as the cellular repair systems were unable to recover the induced toxic insult. Our results showed detection of several CYP1A bands, enlightening an indirect correlation between induction of CYP1A with dose and time of exposure. The decreased expression of CYP1A over prolonged exposure times, along with high toxic concentration, could be related with lethal damage observed on hepatocytes by contrast phase and immunofluorescence analysis. A particular pattern of expression was found for GST isoforms upon AFB1 exposure, identifying each isoform profile two different kind of toxic insult. The 65 KDa and the 49 KDa bands being suggestive for markers of acute and chronic response respectively. Interestingly, apoptosis induction, considered an early lesion to DNA, was found associated with the chronic damage along with the low toxic concentration. The new cell model from S. aurata has been proven to be a useful and valid tool to further investigate the modulated response of liver phase I and II enzymes to AFB1.