The experiment was conducted to evaluate the metabolic activity through MTT (3[4,5-dimethylthiazol-2-y1]-2,5-diphenyltetrazolium bromide) reduction assay of Indian red jungle fowl sperm during cryopreservation process. In addition, MTT test was compared with sperm motility and viability data. Sperm analysis was made at pre-dilution, post -dilution, post-cooling, post-equilibration and post freeze-thawing stages. Pearson's correlation coefficient showed significant (P<0.01) positive association of MTT reduction rate with motility (r=0.85) and viability (r=0.84). A significant (P<0.05) decline was observed in motility, viability and metabolic status after each stage of cryopreservation. The highest (P<0.05) stage-specific percent decrease in motility and viability were observed after freeze-thawing. Nevertheless, stage-specific percent decrease in metabolic status differ (P<0.05) after dilution (5.93 +/- 0.12), cooling (8.41 +/- 0.03), equilibration (19.1 +/- 0.04) and freeze-thawing (30.9 +/- 0.08). The total decrease in motility, viability and metabolic activity after the complete freeze-thawing cycle were calculated as 56%, 47% and 52%, respectively. It is concluded that MTT reduction assay is simple and reliable method to evaluate mitochondrial redox activity in sperm of this threatened species.

1. Birchen and Blue Leonesa are two endangered chicken breeds mainly raised in Curueno Valley in North Spain. The establishment of a germplasm bank to guarantee the preservation of these breeds is needed. However, cockerels from different breeder flocks can show variance in semen cryoresistance. 2. The following work focused on the sperm characterisation and cryopreservation of Birchen and Blue Leonesa cockerels from four different breeders. A total of 30 semen pools were analysed. Besides conventional sperm analysis, including motility by computer-aided sperm analysis (CASA) and DNA fragmentation by TUNEL, the present study tested a double staining method (MitoTracker(TM) Green FM/propidium iodide). This gave simultaneous assessment of plasma and acrosomal and mitochondrial membranes, which were previously validated by SYBR-14/PI, CASA, aniline blue and TUNEL. 3. No significant differences were found among fresh semen variables between breeds and breeders. For post-thawed variables, significant differences (P < 0.05) were found between breeders in sperm viability (58.0 +/- 1.90 breeder D vs. 35.2 +/- 7.41 breeder A, 37.2 +/- 4.09 breeder B and 22.3 +/- 5.92 breeder C) and DNA fragmentation (62.4 +/- 9.91 breeder C vs. 31.8 +/- 7.08 breeder B and 24.5 +/- 5.49 breeder D). The lowest DNA fragmentation values for semen from breeder D birds were coincident with higher integrity of the mitochondrial membrane. 4. The results revealed higher sperm cryoresistance in the cockerels from one of the breeders, possibly due to differences in management system (e.g. diet, housing, control of stress elements and pathogens, reproduction practices or maintenance of genetic diversity). These differences may determine the sperm freezability, and thus the effectiveness of developing a germplasm bank.

The present work aimed to evaluate the chromatin compaction of rooster spermatozoa along the male reproductive tract, and to study the vas deferens lining cells, potentially involved in sperm maturation. Chromomycin A3 (CMA3) was used to determine the chromatin compaction of spermatozoa from testis (T), proximal (including epididymis, V1), intermediate (V2) and distal (V3) vas deferens, and ejaculate (E). Six Birchen Leonesa roosters were used. E was obtained in vivo by dorso-ventral massage. V1, V2 and V3 sperm were obtained post mortem (six pairs of vasa deferentia), by flushing. T was obtained by washing the testes, cut in halves. The fixed cells were stained with CMA3 and propidium iodide for flow cytometry assessment. Results showed higher (P < 0.01) median fluorescence intensity (lower chromatin compaction) of T (693.8 ± 30.2) than V1 (546.3 ± 17.7), V2 (515.1 ± 12.1), V3 (517.6 ± 12.3) and E (491.4 ± 16.1). Regarding the percentage of intensely stained cells, T differs (P < 0.05) from V2, V3 and E, V1 differs (P < 0.05) from V3 and E, while V2, V3 and E do not differ. The histological analysis revealed secretory capacity of the vas deferens. Our findings specified that the transit though the vas deferens results in high percentage of compacted chromatin spermatozoa in E.

A classical chicken semen diluent (Lake's 7.1 diluent) was modified to have lowered osmolalities (ranging from 290 to 410 mOsm/kg). The modified medium with physiological osmolality of 325 mOsm/kg allowed cold storage of fresh semen for several days with very little loss of membrane integrity and motility, while high os-molalities inhibited motility. This modified medium was then used as base for freezing medium to test effects of the type and concentration of cryoprotective agent (CPA), and the cooling rate (CR). A number of CPAs (methylformamide, methylacetamide, dimethylformamide (DMF), dimethylacetamide (DMA), dieth-ylformamide, and propylene glycol) were first compared by freezing semen with 0.6 mol/l of the respective CPA at a cooling rate of 250 degrees C/min. Post-thaw motility and membrane integrity were highest with DMA and DMF. Finally, in more detailed factorial experiments, semen from individual cocks or pooled semen was frozen using CRs of 4, 50, 250, and 440 degrees C/min and DMA concentrations ([DMA]) of 0.4, 0.6, 1.0, and 1.5 mol/l. Straws from each semen sample x treatment combination were divided for semen assessment at three different research groups for sperm motility, membrane integrity, kinked tails, and DNA fragmentation, using microscopy, com-puter assisted motility analysis, and flow cytometry. There were clear effects of both CR and [DMA] and their interaction. CRs 50 and 250 degrees C/min gave best post-thaw sperm performance. Higher DMA concentrations gave better post-thaw membrane integrity, but concentrations above 1.0 mol/l can decrease sperm velocity or even inhibit sperm motility. Therefore [DMA] may best be 0.6-1.0 mol/l at a CR of 50-250 degrees C/min.

Over the last century, several reproductive biotechnologies beyond the artificial incubation of eggs were developed to improve poultry breeding stocks and conserve their genetic diversity. These include artificial insemination (AI), semen storage, diploid primordial germ cell (PGC) methodologies, and gonad tissue storage and transplantation. Currently, AI is widely used for selection purposes in the poultry industry, in the breeding of turkeys and guinea fowl, and to solve fertility problems in duck interspecies crosses for the production of mule ducklings. The decline in some wild game species has also raised interest in reproductive technologies as a means of increasing the production of fertile eggs, and ultimately the number of birds that can be raised. AI requires viable sperm to be preserved in vitro for either short (fresh) or longer periods (chilling or freezing). Since spermatozoa are the most easily accessed sex cells, they are the cell type most commonly preserved by genetic resource banks. However, the cryopreservation of sperm only preserves half of the genome, and it cannot preserve the W chromosome. For avian species, the problem of preserving oocytes and zygotes may be solved via the cryopreservation and transplantation of PGCs and gonad tissue. The present review describes all these procedures and discusses how combining these different technologies allows poultry populations to be conserved and even rapidly reconstituted.

Background Currently, there is a high demand for efficient pig embryo cryopreservation procedures in the porcine industry as well as for genetic diversity preservation and research purposes. To date, vitrification (VIT) is the most efficient method for pig embryo cryopreservation. Despite a high number of embryos survives in vitro after vitrification/warming procedures, the in vivo embryo survival rates after embryo transfer are variable among laboratories. So far, most studies have focused on cryoprotective agents and devices, while the VIT effects on porcine embryonic gene expression remained unclear. The few studies performed were based on vitrified/warmed embryos that were cultured in vitro (IVC) to allow them to re–expand. Thus, the specific alterations of VIT, IVC, and the cumulative effect of both remained unknown. To unveil the VIT-specific embryonic alterations, gene expression in VIT versus (vs.) IVC embryos was analyzed. Additionally, changes derived from both VIT and IVC vs. control embryos (CO) were analyzed to confirm the VIT embryonic alterations. Three groups of in vivo embryos at the blastocyst stage were analyzed by RNA–sequencing: (1) VIT embryos (vitrified/warmed and cultured in vitro), (2) IVC embryos and (3) CO embryos. Results RNA–sequencing revealed three clearly different mRNA profiles for VIT, IVC and CO embryos. Comparative analysis of mRNA profiles between VIT and IVC identified 321, differentially expressed genes (DEG) (FDR < 0.006). In VIT vs. CO and IVC vs. CO, 1901 and 1519 DEG were found, respectively, with an overlap of 1045 genes. VIT-specific functional alterations were associated to response to osmotic stress, response to hormones, and developmental growth. While alterations in response to hypoxia and mitophagy were related to the sum of VIT and IVC effects. Conclusions Our findings revealed new insights into the VIT procedure-specific alterations of embryonic gene expression by first comparing differences in VIT vs. IVC embryos and second by an integrative transcriptome analysis including in vivo control embryos. The identified VIT alterations might reflect the transcriptional signature of the embryo cryodamage but also the embryo healing process overcoming the VIT impacts. Selected validated genes were pointed as potential biomarkers that may help to improve vitrification.

The depth of intravaginal insemination to achieve optimum fertility with frozen-thawed semen is highly species specific in birds and differ even in breed and/or strains of a species. Therefore, study was designed to evaluate the influence of intravaginal insemination depths (2 and 4 cm) on fertility outcome in Indian red jungle fowl. Semen collected from eight mature cocks was pooled, diluted in extender and cooled to 4 degrees C. Glycerol (20%) was added to chilled semen, equilibrated for 10 min and cryopreserved. After 3 days of storage, frozen semen was thawed in water bath at 37 degrees C for 30 s. After glycerol removal, intravaginal Inseminations were performed at the depth of 2 and 4 cm. The no. of fertilized eggs (31.4 +/- 1.6 vs. 27.7 +/- 1.8), fertility rate (65.7 +/- 3.6 vs. 58.8 +/- 4.0), no. of hatched chicks (27.8 +/- 1.9 vs. 23.5 +/- 1.6), hatchability of set eggs (58.8 +/- 4.3 vs. 49.7 +/- 3.2) and hatchability of fertilized eggs (88.4 +/- 2.8 vs. 84.3 +/- 2.2) were recorded higher with intravaginal depth of 4 cm compared to 2 cm. It is concluded that intravaginal insemination at the depth of 4 cm enhances the fertility outcomes of the frozen-thawed Indian red jungle fowl semen.

During cryopreservation sperm encounter oxidative stress due to higher production of ROS molecules and insufficient natural antioxidant defence system. Therefore, present study was designed to identify the effects of various glutathione (GSH) concentrations on Indian red jungle fowl (Gallus gallus murghi) sperm quality and fertility pre-freezing and post-thaw incubation hours. Semen was collected from eight cocks and qualified semen ejaculates having motility >65% were pooled after initial evaluation. Semen was divided in four aliquots, diluted with red fowl extender (1:5) at 37 degrees C having GSH 0 mM (control), 0.1 mM, 0.5 mM and 1.0 mM, cryopreserved and stored at (similar to 196 degrees C) in liquid nitrogen. Semen quality was assessed at post dilution, cooling, equilibration, and freeze-thawing at 0, 2 and 4 h of incubation at 37 degrees C. Sperm motility, plasma membrane integrity, viability, acrosome integrity and mitochondrial function were recorded highest (P < 0.05) with 0.5 mM GSH in extender at post-dilution, cooling, equilibration, freeze-thawing and 0, 2 and 4 h of incubation. Lipid peroxidation in sperm and seminal plasma were recorded lowest (P < 0.05) with 0.5 mM GSH during cryopreservation stages and postthawing incubation. Moreover, antioxidant activities (total antioxidant potential and free radical scavenging capacity) were recorded highest (P < 0.05) in extender having 0.5 mM GSH. Fertility rates were recorded higher (P < 0.05) with 0.5 mM GSH compared to control. It is concluded that 0.5 mM GSH in extender improves sperm structural (sperm viability, plasma membrane integrity and acrosome integrity), functional integrity (motility, mitochondrial function) and fertility parameters of Indian red jungle fowl through enriching antioxidant potential and ameliorating the oxidative stress. (C) 2021 Elsevier Inc. All rights reserved.

The present study investigates the efficacy of dimehtlyformamide (DMF) as a permeable cryoprotectant and its effect on quality and fertility of Indian red jungle fowl sperm. Semen was collected from eight mature roosters, pooled, divided into five aliquots and diluted with red fowl extender having DMF (0%, 4%, 6%, 8% and 10%). Diluted semen samples were cooled from 37 °C to 4 °C, 20% glycerol added to control (0% DMF), equilibrated for 10 min and filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min and plunged into liquid nitrogen. Sperm motility, plasma membrane functionality, viability and acrosome integrity were assessed at post dilution, cooling, equilibration and freeze-thawing stage of cryopreservation. Cryopreservation stages had negative effects (P < 0.05) on semen quality parameters. Percentages of sperm motility, plasma membrane functionality, viability and acrosome integrity were recorded highest in extender having 8% DMF at post-dilution, cooling, equilibration and freeze-thawing stage. Fertility results after artificial insemination were recorded higher (P < 0.05) with 8% DMF compared to 20% glycerol. Dimehtlyformamide (8%) in red fowl extender improves the post thaw semen quality and fertility in Indian red jungle fowl and can be used effectively to avoid the contraceptive effects of glycerol.

Recent reports showed a positive correlation between frozen-thawed rooster sperm DNA integrity and the concentrations of valine in seminal plasma. The present study evaluated the effect of supplementing valine to semen extender for freezing sperm of 2 endangered local Spanish chicken breeds with different sperm cryoresistance: Red Villafranquina (VF) showing low sperm DNA integrity after cryopreservation and Quail Castellana that shows higher DNA integrity. One pool of semen per breed was obtained twice a week for 10 wk (n = 40, 20 per breed). Each pool was divided into 2 fractions. One of these fractions was frozen in presence of valine as additive in the extender (concentration 10 mmol), whereas the other was used as control. The evaluation of the samples before and after freezing-thawing included motility (CASA-Mot system), viability (propidium iodide and SYBR-14), DNA integrity (terminal deoxynucleotidyl transferase dUTP nick end labeling), and fertility rate (percentage of eggs with blastoderm development after artificial insemination). Supplementation of valine increased several motility variables of fresh semen. In VF breed, valine increased percentage of progressive motile sperm (P = 0.025), curvilinear velocity (P = 0.033), straight-line velocity (P 5 0.040), and average path velocity (P = 0.033), whereas progressive motile sperm (P = 0.019), curvilinear velocity (P = 0.006), straight-line velocity (P = 0.003) and average path velocity (P = 0.004) were improved in the Quail Castellana breed. Valine addition increased the DNA integrity of cryopreserved semen (decreased post-thaw DNA fragmentation) in both breeds, with a significant effect (P = 0.002) in VF (36.3% VF-control vs 31%VF-valine). As expected, Quail Castellana cryopreserved sperm control showed higher fertility rate (34.4% 6 12.1) than VF cryopreserved sperm control (16.1% 1 6.2). Supplementing valine to the freezing extender doubled (P 5 0.026) the fertility rate of VF (32.6% 6 12.2) compared with the control (16.1% 1 6.2). In conclusion, supplementation of valine to chicken freezing extenders shows a positive effect on DNA fragmentation and fertilizing ability of frozen-thawed sperm, with a better response in a breed considered as the lowest freezer in our conservatory.

Suite à un appel d’offre du Commissariat Général à la Stratégie et à la Prospective du gouvernement français, l’INRA a analysé les possibilités d’évolution des pratiques et des systèmes agricoles français vers des systèmes de production plus durables conciliant performances productives, économiques, environnementales et sociales. Une méthodologie d’aide à la conception et à l’évaluation de systèmes a été développée à cette occasion. Elle s’appuie sur un répertoire de plus de 200 pratiques agricoles élémentaires dont l’impact a été qualifié sur 35 indicateurs de performances, ainsi que sur une table de compatibilité entre pratiques élémentaires. Deux études de cas fondées sur des modalités différentes d’utilisation de cette méthodologie sont présentées : i) la conception pas à pas de système de production par incorporation progressive de pratiques élémentaires à partir d’une pratique d’entrée à effet majeur, pour accroître l’autonomie alimentaire en production bovine laitière en valorisant la prairie ; ii) l’évaluation à postériori des performances de systèmes en transcrivant sous forme de pratiques élémentaires des leviers d’action et marges de progrès identifiés à dire d’expert pour améliorer la compétitivité de la filière poulet de chair par la mise en place de deux systèmes de production très contrastés mais complémentaires. Les tendances d’évolution des performances des systèmes de production ainsi simulés ou reconstitués sont discutées. Dans les situations d’impact ambivalent d’une pratique sur une performance, le recours au descriptif détaillé des pratiques permet le plus souvent de réduire l’incertitude en prenant en compte les situations associées à chaque type d’impact.

The aim of the present work was to examine the influence of access to pasture in an outdoor housing system on rooster sperm quality and response to cryopreservation and to examine the possible correlation between values for sperm quality variables and welfare indicators. Two groups of Black-barred Andaluza and Red-barred Vasca roosters were housed in an outdoor system, with one group given daily access to a grazing area containing plant species that typically grow on uncultivated Mediterranean land. Semen was collected once per week from each group, and the following sperm quality variables were assessed: sperm volume, appearance, concentration, motility, membrane integrity, acrosome integrity, and morphological abnormalities. In addition, two welfare indicators were examined: the heterophil/lymphocyte (H/L) ratio, and the duration of tonic immobility (TI). Ejaculates from the birds with access to pasture had higher percentages of sperm showing progressive motility (P = 0.019), and returned a higher motility index (P = 0.035). Unexpectedly, the H/L ratio was also higher in these birds. Virtually no differences were seen between the treatment groups with respect to sperm quality after freezing-thawing, although the semen of the Red-barred Vasca birds with access to pasture did show a higher percentage of progressive motility (P = 0.023) than the birds of the same breed with no such access. Significant correlations were detected between the H/L ratio and sperm motility (r = 0.420, P = 0.038), the sperm motility index (r = 0.526, P = 0.002), and progressive motility (r = 0.467, P = 0.003). No differences were seen between the treatment groups with respect to the duration of TI. In conclusion, access to pasture improved fresh sperm motility.

Egg yolk is a good external cryoprotectant of mammalian sperm and some wild bird's sperm, but, at least in domestic breeds of chicken (Gallus gallus domesticus), it may inhibit eventual fertilization of ova when high concentrations are used. We hypothesized that egg yolk can protect the sperm from cryo-induced damages providing adequate fertilization in one phylogenetic wild ancestor of current chicken breeds: the Indian red jungle fowl (Gallus gallus murghi). To test the hypothesis, the present study was designed to evaluate different concentrations of egg yolk in extender in comparison with glycerol. Semen collected from Indian red jungle fowl cocks (n = 8) was cryopreserved using different egg yolk concentrations (10%, 15%, 20% and 25%) or 20% glycerol (control group) following routine protocol of cryopreservation. During cryopreservation, sperm motility (67.5 ± 2.5%), plasma membrane integrity (66.3 ± 2.4%), viability (58.8 ± 1.3%) and acrosomal integrity (60.0.8 ± 2.0%) were recorded highest in an extender with 15% egg yolk compared to other experimental extenders and control at post-dilution, cooling, equilibration and thawing. The no. of fertilized eggs (26.6 ± 0.7, 21.6 ± 1.2), percent fertility (55.9 ± 4.4, 46.5 ± 2.2), no. of hatched chicks (23.6 ± 1.0, 17.2 ± 1.0), percent hatch (49.5 ± 3.2, 37.1 ± 2.5%) and hatchability of the fertile eggs (89.4 ± 2.2, 79.7 ± 3.7) were recorded higher (P < 0.05) with semen cryopreserved with 15% egg yolk compared to control (20% glycerol). It is concluded that 15% egg yolk can be used in cryopreservation protocol of Indian red jungle fowl sperm.

Density-gradients centrifugation (DGC) and filtration columns (FC) are used to separate deformed or dead sperm, debris, and other cells that may negatively affect the fertilizing capacity of sperm in fresh, chilled and frozen/thawed semen. The present study was conducted to evaluate the suitability of DGC (BoviPure®, Percoll® and Accudenz®) and FC (Sephadex G-15®) sperm selection procedures for fresh-extended and cold-stored ram semen by assessment of post-treatment sperm quality variables. Twenty normospermic ejaculates from ten adult Merino rams were used. Sperm concentration of recovered cells was greater (P < 0.001) after BoviPure treatment than other procedures in both fresh and cold semen. With the Sephadex method, there were more desirable values than with use of DGC procedures in several sperm motility variables measured by using the CASA system. In non-refrigerated semen samples, the percentage of progressive sperm motility (%PSM) after Sephadex filtration was greater (P < 0.05) than after BoviPure treatment; the straightline velocity (VSL) value after Sephadex filtration was greater (P < 0.01) than after Accudenz treatment; the amplitude of lateral head displacement (ALH) after Sephadex and Accudenz treatment was less than non-filtered semen (P < 0.001) and after Percoll (P < 0.01) and BoviPure (P < 0.05) treatments. In cold-stored semen samples, the %PSM after Sephadex filtration was greater than non-filtered (P < 0.05) semen and after BoviPure (P < 0.05), Percoll (P < 0.05) and Accudenz (P < 0.001) treatments. It is concluded that Sephadex column filtration can be used to select ram sperm in non-refrigerated and cooled semen, because percentage progressively motile sperm and some other sperm motility characteristics are greater with use of this techniques as compared with use of DGC methods.

It was hypothesized that dimethyleacetamide (DMA) can be used as an alternate to glycerol for cryopreservation of Indian red jungle fowl semen. Four concentrations of DMA (4%, 6%, 8% and 10%) in extender were compared with previously optimized cryopreservation protocol based on 20% glycerol (control) for Indian red jungle fowl. Sperm motility, plasma membrane integrity, viability, and acrosome integrity were assessed at the stage of post-dilution, cooling, equilibration, and freeze-thawing. The whole experiment was repeated/replicated for five times independently. Sperm motility, plasma membrane integrity, viability and acrosome integrity were recorded highest (P < 0.05) at post-dilution, cooling, equilibration, and freeze-thawing in extender having 6% DMA compared to control and other experimental extenders. The highest (P < 0.05) recovery rates of all aforementioned parameters were also recorded in extender having 6% DMA; thus, 6% DMA was further compared with control (20% glycerol) for fertility after artificial insemination. Eggs were collected for five days after artificial insemination with semen cryopreserved in extender containing 6% DMA and control. The higher no. of fertilized eggs, fertility, no. of hatched eggs, hatch (%) and hatchability were recorded with semen cryopreserved in extender having 6% DMA compared to control. It is concluded that 6% DMA maintained higher post-thaw quality and fertility of Indian red jungle fowl semen and is a better replacement of glycerol.

The endogenous peptides and small proteins present in chicken sperm were identified in the context of the characterization of a fertility-diagnostic method based on the use of ICM-MS (Intact Cell Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry). The interpretation and description of these data can be found in a research article, "Intact cell MALDI-TOF MS on sperm: a molecular test for male fertility diagnosis" (Soler et al., 2016) [1], and raw data derived from this analysis have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE: PXD002768. Here, we describe the inventory of all the molecular species identified, along with their biochemical features and functional analysis. This peptide/protein catalogue can be further employed as reference for other studies and reveal that the use of proteomics allows for a global evaluation of sperm cells functions.

The Indian Red Jungle Fowl is a wild native gallus subspecies of Southern Asia. Semen has never been studied in this species. In order to better know the male reproductive capacities, experiments were conducted to study the semen characteristics, impact of ejaculate collection frequencies, and timing of collection on sperm quality parameters. Mean sperm concentration 800 million/mL, total sperm per ejaculate (0.015 billion), motility (63.5 %), live/total sperm (92.4 %), intact acrosome (75.5 %), and plasma membrane integrity (89.2 %) were recorded. Percentage of abnormal sperm (head, mid-piece, and tail) was 8.1 % and recovered mainly mid-piece abnormalities. The motile sperm percentage was positively correlated with intact acrosomes (r = 0.34) and plasma membrane integrity (r = 0.41). Total sperm per ejaculate (billion) was maximum at 72 h of collection followed by 24 and 48 h of collection. Daily and weekly sperm production (billion) was found maximum at 24 h of collection compared to 12, 48, and 72 h of collection. Sperm motility was higher at 24, 48, and 72 h of collection compared to 12 h of collection, but the number of live sperm were higher at 12 h of collection compared to 24, 48, and 72 h. Sperm concentration was better in the morning time, while the values for sperm viability and plasma membrane integrity were higher in the semen collected at evening time. In conclusion, the Indian Red Jungle Fowl shows a semen production quantitatively relatively low for the species as compared to domestic chicken and contrasted parameters of quality. The semen production is affected by the frequency of collection with an optimum for a daily collection preferentially held in the evening period. These results may now be used for artificial insemination and conservation program.

The fertilizing capacity of pure, fresh avian semen may disappear in just half an hour, hindering its successful use in artificial insemination (AI) projects. Longer storage requires the use of infra-physiological temperatures and of semen diluents that help preserve the spermatozoa but that do not interfere with their fertilizing capacity. This study examines the effect on sperm quality of storing red-legged partridge sperm for 3 h at 5°C with 2 different semen extenders: 1) a medium referred to as L&R-84, composed of sodium glutamate, glucose, magnesium acetate, potassium acetate, and polyvinylpyrrolidone, and 2) Lake 7.1 medium, composed of sodium glutamate, glucose, magnesium acetate, potassium citrate, and N,N-Bis(2-hydroxyethyl)taurine (BES). Extending with L&R-84 returned better curvilinear velocity (P < 0.01), straight-line velocity (P < 0.01), average path velocity (P < 0.01), linearity (P < 0.05), straightness (P < 0.05), and wobble (P < 0.05) values, while extending with the Lake 7.1 medium was associated with higher percentages (P < 0.001) of motile sperm. The fertility rate was higher (P < 0.05) when birds were inseminated with L&R-84-extended sperm than with Lake 7.1-extended sperm. The mean number of penetrations of perivitelline layer samples (taken from above the germinal disc) was also higher for the L&R-84-extended sperm (P < 0.05). These results show L&R-84 can be recommended as an extender for red-legged partridge semen to be stored for at least 3 h at 5°C.

Following the bidding by the General Commission for Future Strategy of the French government, INRA has analyzed the different evolutions possible in French practices and agriculture systems towards a more sustainable production that manages to combine productive, economic, environmental and social performances. A method to help design and evaluating the systems was thus developed. This method is based on over 200 basic agricultural practices whose impact was qualified on 35 performance indicators as well as on a compatibility index between these elementary practices. Two studies based on different uses of this method are presented: i) the step by step conception of a production system by progressively incorporating elementary practices from one entry practice with major effect, to increase food autonomy in dairy cattle production by valorizing grasslands; ii) evaluating the system performances a posteriori by transcribing the action levers and progress margins identified by an expert panel into elementary practices in order to improve competition of the broiler industry through the development of two very different but complementary production systems. The tendencies of the simulated or rebuilt performance evolution of these production systems are discussed. When situations present an ambivalent impact of a practice on one performance, using the detailed description of these practices often allows reducing the uncertainty by considering the situations associated with each type of impact.