Improvement Strategies in Ovine Artificial Insemination

P. 30-42Artificial insemination in ram is scarcely widespread comparing with other domestic species. This has been due not only to fertility results being irregular and low but also because of the difficulty in the application of enhancements such as the use of frozen‐thawed sperm. Although there is a lot of information on the use of different options to improve these AI results (such as transcervical application, the use of thawed sperm, etc.) commercial programmes can be classified on two general categories: those using refrigerated semen (15°C) by superficial intracervical deposition (vaginal), and, more restricted, those using thawed sperm by intrauterine deposition (laparoscopy). In the present work, we have summarized our viewpoint on three general research lines for the improvement of AI results in sheep: semen preservation, AI procedures and semen assessment. Briefly, in ram it is necessary to develop a medium term methodology of sperm refrigeration (3–5 days), which would allow the distribution of sperm doses to a widespread area. Nevertheless, it is also necessary to develop an intrauterine transcervical AI technique, which allows thawed semen to be applied by vaginal insemination. Besides, the low predictive value of classic assessment techniques limits the ability to adjust the number of spermatozoa per dose according to its actual fertility

Sperm Subpopulations in Iberian Red Deer Epididymal Sperm and Their Changes Through the Cryopreservation Process

P. 316–327We have applied a statistical protocol based on principal component analysis, clustering methods, and discriminant analysis for the identification of sperm subpopulations in computer-assisted sperm analysis (CASA) data. Samples were obtained from the cauda epididymis of 11 Iberian red deer and cryopreserved following a standard protocol. Motility by CASA was analyzed just after sperm recovery, just before freezing, and after thawing, and eight motility descriptors for each individual spermatozoon were recorded. Sperm viability and acrosomal status were also assessed. Subpopulation analysis was performed in four sequential steps: principal component analysis using the eight motility descriptors; nonhierarchical clustering analysis (k-means) using the first two principal components; hierarchical clustering analysis (UPGMA); and selection of the final number of clusters. Three clusters were obtained for each motility analysis: slow and nonlinear; rapid and linear; and rapid, high ALH, nonlinear. We detected variations in the clusters between treatments (initial, prefreezing and postthawed). Indeed, motility increased and linearity decreased in the prefreezing analysis. A discriminant analysis isolated three descriptors that were used again in the same statistical analysis, giving four clusters that resembled the pattern found in the first classification. We also performed a clustering analysis of the males according to prefreezing/postthawed variation of total motility, viability, and acrosomal status. The proportion of the linear subpopulations in the prefreezing treatment, in both clustering analyses, correlated positively with postthawed viability recovery. Our results show that clustering analysis of CASA data gives useful and practical information that is not obtained by conventional sperm analysis.S

Season effect on genitalia and epididymal sperm from Iberian red deer, roe deer and Cantabrian chamois

P. 1857-1875Seasonality deeply affects the physiology and behavior of many species, and must be taken into account when biological resource banks (BRBs) are established. We have studied the effect of seasonality on many reproductive parameters of free-ranging Iberian red deer, roe deer and Cantabrian chamois, living in Spain. Testicles from hunted animals were collected and sent to our laboratory at different times during the year. We recorded the weight and volume of testis, the weight of the epididymis and its separate parts (caput, corpus, and cauda), the weight of the sperm sample collected from the cauda epididymis, and several sperm parameters (sperm concentration, spermatozoa recovered, motility, HOS test reactivity, acrosomal status, and viability). We studied the data according to several periods, defined accordingly to each species. For red deer, we defined rut (mid-September to mid-October), post-rut (mid-October to mid-December), and non-breeding season (February). For roe deer, they were pre-rut (June), rut (July), post-rut (first fortnight of August), and non-breeding season (September). For chamois: non-breeding season (June to mid-September) and breeding season (October–November). The rut/breeding season yielded significantly higher numbers for almost all parameters. However, in the case of red deer, sperm quality was higher in the post-rut. For roe deer, testicular weight was similar in the pre-rut and in the rut, and sperm quality did not differ significantly between these two periods, although we noticed higher values in the rut. In the case of chamois, sperm quality did not differ significantly from the breeding season, but data distribution suggested that in the non-breeding season there are less males with sperm of good quality. On the whole, we find these results of interest for BRB planning. The best season to collect sperm in this species would be the breeding season. However, post-rut in red deer, pre-rut in roe deer, and non-breeding season in chamois could be used too, because of the acceptable sperm quality, despite the lower quantity salvaged. More in-depth research needs to be carried out on the quality of sperm salvaged at different times of the year in order to confirm these findings.S

A multifaceted provider-centred intervention versus usual care to improve the recognition and diagnosis of depression in primary health care: a hybrid study

Background: The aim of this study was to evaluate the impact of a multifaceted intervention to implement an adapted guideline for the management of depression in primary health care. Methods: A hybrid trial was carried out to determine the effect of a multicomponent provider centred intervention to improve the detection and diagnosis of depression in primary care, as part of the guideline implementation process, and to collect information about barriers and facilitators in a real-world context. Before the multicomponent intervention, a descriptive cross-sectional study was performed to assess the population prevalence of depression in the participating health centres and to detect possible differences. Subsequently, a quasi experimental two-phase study was carried out with a concurrent control group to assess the impact of the multicomponent intervention on the main outcomes (detection of depression, evaluation of its severity and the use of structured methods to support the diagnosis). Results: Nine-hundred seventy-four patients took part in the first phase. According to their clinical records, the prevalence of depression ranged from 7.2% to 7.9%, and there were no significant differences between the health centres scheduled to receive the intervention and those in the control group. In the experimental phase, 797 randomly selected participants received the multicomponent intervention. Adjusted multivariable analysis performed before the implementation revealed no significant differences in depression between the experimental and control groups. However, after the intervention, modest but significant differences were observed, which persisted at 1 year after the intervention. Conclusions: A multicomponent intervention for the implementation of a clinical guideline for the management of depression in primary care produced improvements in the identification of depression and in the degree of severity recorded.Funding for open access charge: Universidad de Málaga

DNA fragmentation assessment by flow cytometry and Sperm–Bos–Halomax (bright‐field microscopy and fluorescence microscopy) in bull sperm

P. 88-98The aim of this study was to find the relationship between fertility (as 90‐day non‐return rates) and DNA fragmentation assessed by two techniques [sperm chromatin structure assay (SCSA) and Sperm–Bos–Halomax (SBH)]. Furthermore, other quality parameters were achieved (motility, morphological abnormalities, cytoplasmic droplets, viability, capacitation and acrosomal and mitochondrial status) and their correlations with fertility were analysed. Bulls were divided into three fertility groups: high [non‐return rate (NRR) ≥ 80], medium (80  40). The results of this study indicate that there is a good correlation between fertility and different parameters of sperm quality (SBH and SCSA parameters, % of spermatozoa with head, neck and total abnormalities, and % of spermatozoa with proximal cytoplasmic droplets) and differences between fertility groups were observed in some of them (SBH and SCSA parameters and % of spermatozoa with head, neck and total abnormalities). In this sense, SBH parameters rendered good correlations with fertility (r = −0.42 using bright light microscope and r = −0.47 with fluorescence). Also, standard deviation of DNA fragmentation index (SD‐DFI) and DFIh (cells with High DNA fragmentation index) showed good correlations with fertility (r = −0.41 and r = −0.29). No correlations were observed between SCSA and SBH parameters. A multiple regression shows that four parameters (% of proximal cytoplasmic droplets, % of intact acrosomes in total population, SD‐DFI and percentage of fragmented DNA detected by bright light microscope) present a good predictive value of the fertility of sperm samples (r2 = 0.34, p < 0.001

Effects of cryopreservation on head morphometry and its relation with chromatin status in brown bear (Ursus arctos) spermatozoa

P. 1498-1506The Cantabrian brown bear (Ursus arctos) is a highly endangered species in Spain and basic studies are necessary in order to bank its germplasm. Sperm heads are mainly made up of chromatin, thus their shape depends partly on chromatin structure. Thawed semen from 10 bears was used to analyze chromatin status by sperm chromatin structure assay (SCSA) and head morphometry by the computer-assisted sperm morphology assessment (CASMA) system. Morphometry was analyzed before and after freezing–thawing in order to evaluate the effects of cryopreservation on sperm heads. Each spermatozoon was measured for four primary parameters (length, L; width, W; area, A; perimeter, P) and derived parameters (ellipticity: L/W, circularity: 4πA/P2, elongation: (L − W)/(L + W), regularity: πLW/4A). All the derived parameters significantly differed between bears. Likewise, cryopreservation affected head morphometry by reducing its size. Clustering based on morphometric parameters separated three subpopulations, one of them being significantly more influenced by the cryopreservation process. We obtained high correlations between head morphometry and SCSA parameters: standard deviation of DNA fragmentation index (SD-DFI) was correlated with perimeter and area (r = 0.75 and r = 0.62, respectively) and DFIm and DFIt (moderate and total DNA fragmentation index) were correlated with perimeter (r = 0.65 and r = 0.67, respectively). Nevertheless, classification of males according to SCSA or head morphometry did not completely agree so the two assays might explain male variability differently. We conclude that cryopreservation affected morphometry at least in a subset of spermatozoa. These results might improve future application of sperm banking techniques in this species.S

A pilot study on post-thawing quality of Iberian red deer spermatozoa (epididymal and electroejaculated) depending on glycerol concentration and extender osmolality

P. 1165-1172The optimization of cryopreservation extenders is a fundamental issue for adequately performing germplasm banking on wild species. We have tested two glycerol concentrations (4 and 8%), and three extender osmolalities (320, 380 and 430 mOsm/kg; before adding cryoprotectants), for cryopreservation of epididymal and ejaculated sperm samples from Iberian red deer. All the extenders were based on Tes–Tris and fructose (for osmolality adjustment), and complemented with 20% egg yolk. Epididymal and ejaculated sperm samples were obtained from the cauda epididymis (post-mortem) and using electroejaculation, respectively. Samples were diluted 1:1 with each extender and equilibrated for 2 h at 5 °C. Then, they were diluted down to 100 × 106 sperm/mL and frozen at −20 °C/min. Post-thawed samples were assessed for motility (CASA), HOS test, proportion of swollen (osmotically challenged) cells in the untreated sample, viability and acrosomal status. For epididymal samples, 8% glycerol rendered a slightly higher proportion of intact acrosomes on viable spermatozoa than 4%; regarding extender osmolality, 380 and 430 mOsm/kg rendered higher motility results, and the 430 mOsm/kg yielded the lowest proportion of swollen spermatozoa. For ejaculated samples, 4% glycerol yielded more viable spermatozoa than 8%; for extender osmolality, 320 mOsm/kg rendered the highest percentages of progressively motile and viable spermatozoa, although 380 mOsm/kg extender was not significantly different. These results show that sample source influences extender suitability, and that extenders should be isoosmotic or rather slightly hyperosmotic. Future studies should test multiple glycerol concentrations and extender osmolalities in order to adjust them to these kinds of sample.S

The World Spider Trait database : a centralised global open repository for curated data on spider traits

Publisher Copyright: © The Author(s) 2021. Published by Oxford University Press.Spiders are a highly diversified group of arthropods and play an important role in terrestrial ecosystems as ubiquitous predators, which makes them a suitable group to test a variety of eco-evolutionary hypotheses. For this purpose, knowledge of a diverse range of species traits is required. Until now, data on spider traits have been scattered across thousands of publications produced for over two centuries and written in diverse languages. To facilitate access to such data, we developed an online database for archiving and accessing spider traits at a global scale. The database has been designed to accommodate a great variety of traits (e.g. ecological, behavioural and morphological) measured at individual, species or higher taxonomic levels. Records are accompanied by extensive metadata (e.g. location and method). The database is curated by an expert team, regularly updated and open to any user. A future goal of the growing database is to include all published and unpublished data on spider traits provided by experts worldwide and to facilitate broad cross-taxon assays in functional ecology and comparative biology. Database URL:https://spidertraits.sci.muni.cz/.Peer reviewe

The World Spider Trait database: a centralized global open repository for curated data on spider traits

Spiders are a highly diversified group of arthropods and play an important role in terrestrial ecosystems as ubiquitous predators, which makes them a suitable group to test a variety of eco-evolutionary hypotheses. For this purpose, knowledge of a diverse range of species traits is required. Until now, data on spider traits have been scattered across thousands of publications produced for over two centuries and written in diverse languages. To facilitate access to such data, we developed an online database for archiving and accessing spider traits at a global scale. The database has been designed to accommodate a great variety of traits (e.g. ecological, behavioural and morphological) measured at individual, species or higher taxonomic levels. Records are accompanied by extensive metadata (e.g. location and method). The database is curated by an expert team, regularly updated and open to any user. A future goal of the growing database is to include all published and unpublished data on spider traits provided by experts worldwide and to facilitate broad cross-taxon assays in functional ecology and comparative biology.Fil: Pekár, Stano. Masaryk University; República ChecaFil: Wolff, Jonas O. University of Greifswald; AlemaniaFil: Cernecká, L'udmila. Slovak Academy of Sciences; ArgentinaFil: Birkhofer, Klaus. Brandenburgische Technische Universität Cottbus; AlemaniaFil: Mammola, Stefano. University of Helsinki; FinlandiaFil: Lowe, Elizabeth C.. Macquarie University; AustraliaFil: Fukushima, Caroline S.. University of Helsinki; FinlandiaFil: Herberstein, Marie E.. Macquarie University; AustraliaFil: Kucera, Adam. Masaryk University; República ChecaFil: Buzatto, Bruno A.. University of Western Australia; AustraliaFil: Djoudi, El Aziz. Brandenburgische Technische Universität Cottbus; AlemaniaFil: Domenech, Marc. Universidad de Barcelona; EspañaFil: Enciso, Alison Vanesa. Fundación Protectora Ambiental Planadas Tolima; ColombiaFil: Piñanez Espejo, Yolanda María Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; ArgentinaFil: Febles, Sara. No especifíca;Fil: García, Luis F. Universidad de la República; UruguayFil: Gonçalves Souza, Thiago. Universidad Federal Rural Pernambuco; BrasilFil: Isaia, Marco. Università di Torino; ItaliaFil: Lafage, Denis. Universite de Rennes I; FranciaFil: Líznarová, Eva. Masaryk University; República ChecaFil: Macías Hernández, Nuria. Universidad de La Laguna; EspañaFil: Fiorini de Magalhaes, Ivan Luiz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Malumbres Olarte, Jagoba. Universidade Dos Açores; PortugalFil: Michálek, Ondrej. Masaryk University; República ChecaFil: Michalik, Peter. ERNST MORITZ ARNDT UNIVERSITÄT GREIFSWALD (UG);Fil: Michalko, Radek. No especifíca;Fil: Milano, Filippo. Università di Torino; ItaliaFil: Munévar, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; ArgentinaFil: Nentwig, Wolfgang. University of Bern; SuizaFil: Nicolosi, Giuseppe. Università di Torino; ItaliaFil: Painting, Christina J. No especifíca;Fil: Pétillon, Julien. Universite de Rennes I; FranciaFil: Piano, Elena. Università di Torino; ItaliaFil: Privet, Kaïna. Universite de Rennes I; FranciaFil: Ramirez, Martin Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Ramos, Cândida. No especifíca;Fil: Rezác, Milan. No especifíca;Fil: Ridel, Aurélien. Universite de Rennes I; FranciaFil: Ruzicka, Vlastimil. No especifíca;Fil: Santos, Irene. No especifíca;Fil: Sentenská, Lenka. Masaryk University; República ChecaFil: Walker, Leilani. No especifíca;Fil: Wierucka, Kaja. Universitat Zurich; SuizaFil: Zurita, Gustavo Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; ArgentinaFil: Cardoso, Pedro. No especifíca

DNA fragmentation assessment by flow cytometry and Sperm-Bos-Halomax (bright-field microscopy and fluorescence microscopy) in bull sperm

The aim of this study was to find the relationship between fertility (as 90-day non-return rates) and DNA fragmentation assessed by two techniques [sperm chromatin structure assay (SCSA) and Sperm–Bos–Halomax (SBH)]. Furthermore, other quality parameters were achieved (motility, morphological abnormalities, cytoplasmic droplets, viability, capacitation and acrosomal and mitochondrial status) and their correlations with fertility were analysed. Bulls were divided into three fertility groups: high [non-return rate (NRR) ≥ 80], medium (80  40). The results of this study indicate that there is a good correlation between fertility and different parameters of sperm quality (SBH and SCSA parameters, % of spermatozoa with head, neck and total abnormalities, and % of spermatozoa with proximal cytoplasmic droplets) and differences between fertility groups were observed in some of them (SBH and SCSA parameters and % of spermatozoa with head, neck and total abnormalities). In this sense, SBH parameters rendered good correlations with fertility (r = −0.42 using bright light microscope and r = −0.47 with fluorescence). Also, standard deviation of DNA fragmentation index (SD-DFI) and DFIh (cells with High DNA fragmentation index) showed good correlations with fertility (r = −0.41 and r = −0.29). No correlations were observed between SCSA and SBH parameters. A multiple regression shows that four parameters (% of proximal cytoplasmic droplets, % of intact acrosomes in total population, SD-DFI and percentage of fragmented DNA detected by bright light microscope) present a good predictive value of the fertility of sperm samples (r2 = 0.34, p < 0.001).This work was supported in part by ABEREKIN S.A.Peer reviewe