Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative stress generated through the entire process of cryopreservation. Recently, artificial or natural nanovesicles including liposomes and exosomes, respectively, have shown regenerative capabilities to repair damaged sperm during the freeze-thaw process. Exosomes possess a potential pleiotropic effect because they contain antioxidants, lipids, and other bioactive molecules regulating and repairing spermatozoa. In this review, we highlight the current strategies of using nanoparticles and nanovesicles (liposomes and exosomes) to combat the cryoinjuries associated with semen cryopreservation.

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The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation

Antimicrobial activity of silver-carbon nanoparticles on the bacterial flora of bull semen

Equine Reproductive Physiology, Breeding and Stud Management

Advances in applications of color Doppler ultrasonography in the andrological assessment of domestic animals: A review.

Assessment of correlations and concentrations of salivary and plasma steroids, testicular morphometry, and semen quality in different climatic conditions in goats

This study investigated correlations among ram semen quality parameters, testicular vascularity and echotexture. Measurements were performed biweekly for a year, before (BE) and after ejaculation (AE), on seven Chios breed rams [three rams (group A) and four rams (group B) of 2-6 and 9-13 years old, respectively], considering breeding (1) and non-breeding period (2). Hemodynamics [Pulsatility Index (PI), Resistive Index (RI)] were evaluated in each testis. Images of testicular parenchyma were analysed to evaluate echogenicity [Mean Value (MV), Contrast (Con), Gray Value Distribution (GVD), Run Length Distribution (RunLD), Long Run Emphasis (LRunEm)]. Semen volume (V), concentration (C), kinetics (CASA), morphology (Sperm Blue), viability (Eosin-Nigrosin) and DNA fragmentation (Acridine Orange) were assessed. In group A/2, semen V was correlated to left testis PI and right testis RI, BE (r = 0.4, p = 0.013/r = 0.6, p < 0.001, respectively), and to right testis PI and RI, AE (r = 0.5, p < 0.001). In A/1/AE, semen C was correlated to left testis PI (r = -0.5, p = 0.003). In group A/1/BE, RI of both testes was correlated to total and tail abnormalities (left: r = -0.5, p = 0.008/ p = 0.004, respectively - right: r = -0.4/r = -0.5, p = 0.01/ p = 0.008, respectively). In group B/1/BE, sperm tail and total abnormalities were correlated to left testis Con (r = 0.5, p = 0.02/ p < 0.001, respectively), whereas in A/2/BE, they were correlated to left testis LRunEm (r = -0.8/r = -0.7, p < 0.001, respectively). In group A/1/BE, DNA damage was correlated to right testis LRunEm (r = -0.6, p < 0.001) and tail abnormalities to left testis PI (r = -0.5, p = 0.03). In group B/1/AE, right testis LRunEm, GVD, RunLD, were correlated to sperm DNA integrity (r = -0.9, p < 0.001). Immotile spermatozoa and viability were correlated to left testis Con in group B/2/AE (r = -0.6/r = 0.6, p < 0.001) and to right testis MV in group A/1/AE (r = -0.6/r = 0.6, p < 0.001). In conclusion, alterations in both ram testes blood flow and echotexture seem to be moderately correlated with sperm assessed parameters, depending on season and ram age.

Effects of testicular hemodynamic and echogenicity changes on ram semen characteristics

Impact of old age and season on Chios ram semen quality

This study examined the effect of Fe3O4 nanoparticles on boar semen. Beltsville thawing solution without antibiotics was used to extend ejaculates from 5 boars (4 ejaculates/boar). Semen samples of control group (C) and group with Fe3O4 (Fe; 0.192 mg/mL semen) were incubated under routine boar semen storage temperature (17 °C) for 0.5 h and nanoparticles were removed by a magnetic field. Before and after treatment, aliquots of all groups were cultured using standard microbiological methods. The samples after treatment were stored (17 °C) for 48 h and sperm parameters (computer-assisted sperm analyzer (CASA) variables; morphology; viability; hypo-osmotic swelling test (HOST); DNA integrity) were evaluated at storage times 0, 24, 48 h. Semen data were analyzed by a repeated measures mixed model and microbial data with Student’s t-test for paired samples. Regarding CASA parameters, Fe group did not differ from C at any time point. In group C, total motility after 24 h and progressive motility after 48 h of storage decreased significantly compared to 0 h. In group Fe, linearity (LIN) after 48 h and head abnormalities after 24 h of storage increased significantly compared to 0 h. The microbiological results revealed a significant reduction of the bacterial load in group Fe compared to control at both 24 and 48 h. In conclusion, the use of Fe3O4 nanoparticles during semen processing provided a slight anti-microbiological effect with no adverse effects on sperm characteristics.

https://www.mdpi.com/2079-4991/10/8/1568/pdf

Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

Η κρυοσυντήρηση του σπέρματος του κριού αποσκοπeί στη γeνeτική βeλτίωση μέσω της τeχνητής σπeρματέγχυσης, στην άρση των γeωγραφικών πeριορισμών, στην προστασία απeιλούμeνων μe eξαφάνιση φυλών και στη διατήρηση της βιοποικιλότητας. Η διαδικασία κατάψυξης όμως, eπιφέρeι δομικές, βιοχημικές και λeιτουργικές μeταβολές στα σπeρματοζωάρια. Συγκeκριμένα, βλάπτονται οι κυτταρικές μeμβράνeς και αυξάνeται η διαπeρατότητά τους, μeταβάλλeται η δομή της χρωματίνης τους και προκαλeίται υπeροξeίδωση καθώς παράγονται δραστικές μορφές οξυγόνου. Οι αλλαγές αυτές, eπηρeάζουν αρνητικά τη γονιμοποιητική ικανότητα των σπeρματοζωαρίων του κριού και την πρώιμη eμβρυική ανάπτυξη. Στη βιβλιογραφία eντοπίζονται πολυάριθμeς eπιστημονικές προσπάθeιeς βeλτίωσης της γονιμότητας του κρυοσυντηρημένου σπέρματος του κριού. Πρωταρχικό και κοινό σημeίο διeρeύνησης, υπήρξe η σύνθeση των αραιωτικών μέσων. Τα αραιωτικά του σπέρματος πρέπeι να έχουν, μeταξύ άλλων, κατάλληλη ρυθμιστική ικανότητα και pΗ ώστe να προστατeύουν τα σπeρματοζωάρια από την οσμωτική και τη θeρμική καταπόνηση. Υπό αυτό το πρίσμα, έγινe προσθήκη και μeλέτη πληθώρας ουσιών μe ποικίλeς ιδιότητeς και αποτeλέσματα. Μeταξύ αυτών, χρησιμοποιήθηκαν κρυοπροστατeυτικά που διαπeρνούν (γλυκeρίνη, διμeθυλο-σουλφοξeίδιο) ή όχι (κρόκος αυγού, αποβουτυρωμένο γάλα) την κυτταρική μeμβράνη, σάκχαρα (γλυκόζη, λακτόζη, τρeχαλόζη, ραφινόζη), άλατα (κιτρικό νάτριο, κιτρικό οξύ) και αντιοξeιδωτικά (αμινοξέα, βιταμίνeς, ένζυμα). Άλλοι παράγοντeς που μeλeτώνται eκτeνώς, αφορούν στη θeρμοκρασία και στο χρόνο συντήρησης στα eπιμέρους στάδια της κατάψυξης, στο ρυθμό πτώσης της θeρμοκρασίας και στο πρωτόκολλο αναθέρμανσης. Τα αποτeλέσματα των eρeυνών eίναι eνθαρρυντικά, όχι μόνο ως προς τις διαδικασίeς κατάψυξης/αναθέρμανσης του σπέρματος, αλλά και ως προς τις ωφέλιμeς ιδιότητeς των προσθeτικών. Παρά την πρόοδο που eπιτeύχθηκe μέχρι σήμeρα, eξακολουθούν να υφίστανται κeνά, μέχρις ότου eπιτeυχθeί η διeυρυμένη χρήση του κρυοσυντηρημένου σπέρματος του κριού στην αναπαραγωγική διαχeίριση των ποιμνίων.

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Current status and advances in ram semen cryopreservation

Contents
The objective of this study was to investigate the quality of frozen-thawed semen from different bull breeds. Commercial frozen-thawed bull semen samples (26 per breed, 130 totally) of five breeds (Holstein [Η], Brown Swiss [BS], Limousin [L], Belgian Blue [BB], Blonde d' Aquitaine [BA]) were used. After thawing, each semen sample was subjected to thermal resistance test (TR) for 0.5 and 1 hr at 38°C and hypo-osmotic swelling test (HOST) for 1 hr at 150 mOsm at 37°C. Additionally, all samples were evaluated at times 0 hr (thawing), 0.5 hr (TR), 1 hr (TR) for kinetics by CASA [progressive, immotile, rapid, medium, slow moving spermatozoa, curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), linearity (LIN), straightness (STR), beat cross-frequency (BCF), amplitude of lateral head displacement (ALH), wobble (WOB)]. Moreover, directly after thawing, all semen samples were evaluated for morphometry, morphology, viability and DNA fragmentation. Statistical analysis was conducted using a mixed model for repeated measures. The results showed (a) higher VCL after thawing in H, L breeds compared to BB and BA, (b) higher VAP after thawing in L compared to BB, BA, (c) higher values of progressive spermatozoa after TR in H, BS compared to BB, BA, (d) higher values of rapid spermatozoa after thawing and 0.5 hr of TR in H, BS, L compared to BB, BA, (e) lower viability in BA after thawing compared to H, BS, BB, (f) lower morphological abnormalities in H compared to L, BB, (g) higher head length in Η compared to BB. No significant differences were observed in the results from HOST and DNA fragmentation between breeds. In conclusion, quality characteristics of frozen-thawed bull semen are dependent on the breed. Frozen semen from BB and BA breeds should be handled more carefully after thawing, as it is more sensitive to stress.

Aikaterini Ntemka

Papers

Effects of testicular hemodynamic and echogenicity changes on ram semen characteristics

Impact of old age and season on Chios ram semen quality

Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

Current status and advances in ram semen cryopreservation

Breed Differences of Bull Frozen-Thawed Semen