Assessment of Sperm Viability and Computer-Assisted Motility Analysis in Budgerigars (Melopsittacus undulatus): Effect of Several In Vitro Processing Conditions.

In order to preserve endangered psittacine species, more basic and applied research in reproductive biology is required. Assisted reproductive technologies such as artificial insemination play an important role in parrots species conservation programs to overcome the problem of infertile eggs and male infertility. The aim of this study was to define an effective in vitro protocol in order to standardize the sperm quality evaluation in psittacines, studying Melopsittacus undulatus as model species. Semen was collected from twenty adult males by massage technique from May to June. Sperm concentration was measured by the spectrophotometric method. Sperm quality (sperm membrane integrity (SMI), motility, and kinetic parameters) was assessed on fresh semen. Three different experimental protocols were performed to compare the effects of various processing conditions on SMI, motility, and kinetic parameters. In protocol 1, test was performed by Lake extender with three different pH, 7.4 versus 8.2 versus 8.4, and two different equilibration temperatures after dilution of fresh semen (4°C versus 25°C). In protocol 2, two dilution rates of semen after collection were valuated, 1 : 3 versus 1 : 4, as well as three different semen storage temperatures (4°C versus 25°C versus 38°C) before sperm motility analysis with the computer-assisted sperm analysis (CASA). In protocol 3, two different Makler chamber temperatures (38 versus 41°C) during motility analysis were tested. A significant progressive improvement in spermatozoa motility and kinetic parameters was registered with pH 8.4. Progressive motility and all kinetic parameters were higher at 4°C equilibration temperature. Straightness (STR) kinetic parameter was better with 1 : 4 dilution rate. Total motile sperm was higher in 41°C Makler chamber. In this study, for the first time, the effects of different processing protocols on psittacines seminal quality analysis were investigated. Significant differences conditioning the effectiveness of analysis protocols have been described.

Concentration dependent effect of dimethylacetamide and N-methylacetamide on the quality and fertility of cryopreserved chicken semen.

The aim of this study was to compare the effect of two permeant-cryoprotectants, dimethylacetamide (DMA) and N-methylacetamide (NMA) used at different concentrations (0%, 2%, 4%, 6%) on the quality and fertility of post-thaw rooster semen. Ejaculates were processed in 7 treatments: Lake pre-freezing+0.1 M trehalose (LPF-T) (control treatment), LPF-T+2% DMA, LPF-T+4% DMA, LPF-T+6% DMA, LPF-T+2% NMA, LPF-T+4% NMA, LPF-T+6% NMA. Sperm quality [sperm membrane integrity (SMI), motility and kinetic parameters] was assessed before and after cryopreservation. Fertility and embryo viability were recorded. Increasing both DMA and NMA concentration from 2 to 6% improved SMI, total motile sperm, progressive motile sperm (PMS), VCL, VSL and VAP values. PMS recovery rates were significantly the highest in 6% DMA, 4% NMA and 6% NMA treatments. Semen cryopreserved with DMA produced the best fertility and embryo viability at 6%; progressive lower values were recorded at lower concentrations, with no viable embryos at 2%. Semen cryopreserved with NMA showed the best fertility values at 2% and lower values were recorded at higher concentrations; live embryos were found in all NMA treatments. Finally, NMA and DMA showed a similar positive concentration dependent effect of the quality of cryopreserved semen. NMA, not DMA, provided the highest fertility and embryo viability values at the lowest 2%. Therefore, the use of NMA is recommended in order to reduce the cryoprotectant concentration, with a concomitant reduction in the risk of toxicity, providing at the same time the adequate cryoprotective action to obtain viable embryos after artificial insemination of cryopreserved chicken semen.

Reproductive parameters in some captive-bred cockatoo species (genus Cacatua and Eolophus).

BACKGROUND: Cockatoo reproduction in captivity supplies a powerful tool to limit the economic motivation to capture endangered species from the wild; nevertheless, scientific data about reproductive parameters in cockatoos are very rare. The aim of the present work was to investigate the reproductive performance of different species of the Cacatua genus reared in the same facility to evaluate adaptability to captive breeding and to identify the main problems in ex situ conservation of some cockatoo species. METHODS: Data of 28 eggs from 19 reproductive pairs from 9 cockatoo species were analysed. Statistical analysis was carried out by SAS NPAR1WAY procedure: species was considered source of variation. RESULTS: Species effect does not significantly influence reproductive variables; differences were recorded in eggs fertility and embryo liveability. Bird adaptive ability to captive breeding has been described through reproductive parameters. CONCLUSION: Our results show the importance and the maintenance of natural species-specific behaviours and habits, and they underline the relevance of data collection about reproductive performance in endangered species kept in captivity to improve breeding management in conservation programmes.

Physical Parameters and Fatty Acids Profiles in Milanino, Mericanel Della Brianza, Valdarnese Bianca and Commercial Hybrids (Gallus Gallus Domesticus) Table Eggs.

The aim of the present study is to investigate the physical parameters and fatty acid composition and related nutritional parameters of market-procured table eggs from Milanino, Mericanel della Brianza and Valdarnese Bianca hens compared to two commercial hybrid strains' eggs to determine characterizing quality traits for traditional breeds conservation and valorization through high quality niche products. Fifty-four market eggs by three traditional breeds (Mericanel della Brianza-MRC; Milanino-MLN; and Valdarnese Bianca-VLD) and two commercial hybrid strains (Commercial Hybrid Brown-CHB; Commercial Hybrid White-CHW) have been analyzed-physical parameters, fatty acids profile and atherogenic and thrombogenic indexes were investigated. A General Linear Model-GLM was applied to data analysis with breed and genetic origin (traditional breed-TRD; commercial hybrid-HYB) as sources of variation. Two Principal Component Analyses (PCA) were carried out with physical parameters and fatty acid parameters as variables. Eggs produced by traditional breeds MRC MLN and VLD differentiate from eggs produced by commercial hybrids CHB and CHW in physical and chemical parameters (fatty acids parameters). The nutritional value of the traditional eggs has been demonstrated to be higher considering the yolk content, the PUFA fraction, the more favorable n6/n3 ratio and the atherogenic and thrombogenic indexes. Commercial layers' eggs revealed their higher commercial value based on weight, albumen content and percentage of edible content.

Effect of dimethylacetamide and N-methylacetamide on the quality and fertility of frozen/thawed chicken semen.

The aim was to compare the effect of dimethylacetamide (DMA) and N-methylacetamide (NMA) concentrations on the quality and fertility of post-thaw chicken semen. Ejaculates were obtained from 30 Hi-Line White roosters and processed according to the following treatments: lake pre-freezing extender + 0.1 M trehalose (LPF-T) + 6% DMA (control treatment), LPF-T + 9% DMA, LPF-T + 6% NMA, and LPF-T + 9% NMA. Sperm quality (viability, motility, and kinetic traits) was assessed before and after cryopreservation. A total of 15 laying hens per treatment were inseminated to assess fertility and embryo viability. Sperm cryopreserved in presence of DMA had significantly better in vitro quality compared to NMA, showing the highest proportion of viable and progressive motile sperm recovered after thawing. Furthermore, proportion of progressive motile sperm and the VCL, LIN, ALH, and WOB mean values were significantly improved in semen samples frozen/thawed with 6% compared to 9% cryoprotectant concentration. However, the best cryoprotective action on sperm quality played by DMA and the lowest cryoprotectant concentration did not translate into a concomitant advantage in in vivo semen fertility that showed no differences between cryoprotectant and cryoprotectant concentration treatments. Finally, the cryoprotectant DMA and NMA showed an opposite effect on embryo viability in comparison with the effect played on in vitro semen quality, being NMA more efficient than DMA on preserving viable embryos. The present results suggest the urgency to further decrease the cryoprotectant concentration in poultry semen freezing procedures and to assess the specific toxic effect of cryoprotectant on sperm integrity, fertility, and embryo development.