Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth

The cryopreservation of rabbit semen is a valuable strategy for genetic resource preservation and efficient artificial insemination, but outcomes remain inconsistent, partly due to variations in sperm concentration per dose. This study aimed to evaluate the in vivo effects of different sperm concent...

Full description

Saved in:
Bibliographic Details
Main Authors: Michele Di Iorio, Giusy Rusco, Fabrizio Lauriola, Emanuele Antenucci, Alessandra Roncarati, Silvia Cerolini, Michele Schiavitto, Nicolaia Iaffaldano
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Veterinary Sciences
Subjects:
Online Access:https://www.mdpi.com/2306-7381/12/7/678
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849713402646102016
author Michele Di Iorio
Giusy Rusco
Fabrizio Lauriola
Emanuele Antenucci
Alessandra Roncarati
Silvia Cerolini
Michele Schiavitto
Nicolaia Iaffaldano
author_facet Michele Di Iorio
Giusy Rusco
Fabrizio Lauriola
Emanuele Antenucci
Alessandra Roncarati
Silvia Cerolini
Michele Schiavitto
Nicolaia Iaffaldano
author_sort Michele Di Iorio
collection DOAJ
description The cryopreservation of rabbit semen is a valuable strategy for genetic resource preservation and efficient artificial insemination, but outcomes remain inconsistent, partly due to variations in sperm concentration per dose. This study aimed to evaluate the in vivo effects of different sperm concentrations (15, 25, 35, 55, and 75 million per straw) on fertility, prolificacy, and offspring growth in nulliparous and multiparous does. A total of 384 rabbit females were inseminated using frozen–thawed semen, and their reproductive performance was compared with fresh semen. Fertility and kindling rates varied with sperm concentration and parity: nulliparous does showed the highest fertility at 15 million sperm/straw (84.4%), while multiparous does reached peak values at 25–55 million/straw (78.1–81.3%). Litter size and live-born kits were consistently higher in multiparous than in nulliparous does. Offspring body weight at 19 and 60 days was influenced by both sperm concentration and maternal parity, with better growth generally observed in multiparous groups. Weaning success remained high across all groups. Our results indicate that sperm concentrations ranging from 15 to 35 × 10<sup>6</sup>/straw are the most suitable for cryopreservation, as they maintain high fertility, prolificacy, and offspring growth, comparable to fresh semen. These results confirm that optimizing sperm concentration during cryopreservation improves reproductive efficiency and that tailoring insemination strategies to the physiological status of the female enhances outcomes. The results provide useful recommendations for improving cryopreservation techniques in rabbit breeding programs.
format Article
id doaj-art-db9848460f41453f94f3c73eebef9321
institution DOAJ
issn 2306-7381
language English
publishDate 2025-07-01
publisher MDPI AG
record_format Article
series Veterinary Sciences
spelling doaj-art-db9848460f41453f94f3c73eebef93212025-08-20T03:13:58ZengMDPI AGVeterinary Sciences2306-73812025-07-0112767810.3390/vetsci12070678Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring GrowthMichele Di Iorio0Giusy Rusco1Fabrizio Lauriola2Emanuele Antenucci3Alessandra Roncarati4Silvia Cerolini5Michele Schiavitto6Nicolaia Iaffaldano7Department of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis snc, 86100 Campobasso, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis snc, 86100 Campobasso, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis snc, 86100 Campobasso, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis snc, 86100 Campobasso, ItalySchool of Biosciences and Veterinary Medicine, University of Camerino, Via Circonvallazione 93/95, 62024 Matelica, ItalyDepartment of Veterinary Medicine and Animal Science, University of Milan, Via dell’Università 6, 26900 Lodi, ItalyItalian Rabbit Breeders Association (ANCI-AIA), Volturara Appula, 71030 Foggia, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis snc, 86100 Campobasso, ItalyThe cryopreservation of rabbit semen is a valuable strategy for genetic resource preservation and efficient artificial insemination, but outcomes remain inconsistent, partly due to variations in sperm concentration per dose. This study aimed to evaluate the in vivo effects of different sperm concentrations (15, 25, 35, 55, and 75 million per straw) on fertility, prolificacy, and offspring growth in nulliparous and multiparous does. A total of 384 rabbit females were inseminated using frozen–thawed semen, and their reproductive performance was compared with fresh semen. Fertility and kindling rates varied with sperm concentration and parity: nulliparous does showed the highest fertility at 15 million sperm/straw (84.4%), while multiparous does reached peak values at 25–55 million/straw (78.1–81.3%). Litter size and live-born kits were consistently higher in multiparous than in nulliparous does. Offspring body weight at 19 and 60 days was influenced by both sperm concentration and maternal parity, with better growth generally observed in multiparous groups. Weaning success remained high across all groups. Our results indicate that sperm concentrations ranging from 15 to 35 × 10<sup>6</sup>/straw are the most suitable for cryopreservation, as they maintain high fertility, prolificacy, and offspring growth, comparable to fresh semen. These results confirm that optimizing sperm concentration during cryopreservation improves reproductive efficiency and that tailoring insemination strategies to the physiological status of the female enhances outcomes. The results provide useful recommendations for improving cryopreservation techniques in rabbit breeding programs.https://www.mdpi.com/2306-7381/12/7/678rabbit spermatozoacryopreservation protocolin vivo assessmentfertilitypostnatal development
spellingShingle Michele Di Iorio
Giusy Rusco
Fabrizio Lauriola
Emanuele Antenucci
Alessandra Roncarati
Silvia Cerolini
Michele Schiavitto
Nicolaia Iaffaldano
Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
Veterinary Sciences
rabbit spermatozoa
cryopreservation protocol
in vivo assessment
fertility
postnatal development
title Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
title_full Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
title_fullStr Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
title_full_unstemmed Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
title_short Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
title_sort validating sperm concentration in rabbit cryopreservation protocol implications for fertility litter size and offspring growth
topic rabbit spermatozoa
cryopreservation protocol
in vivo assessment
fertility
postnatal development
url https://www.mdpi.com/2306-7381/12/7/678
work_keys_str_mv AT michelediiorio validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT giusyrusco validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT fabriziolauriola validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT emanueleantenucci validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT alessandraroncarati validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT silviacerolini validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT micheleschiavitto validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth
AT nicolaiaiaffaldano validatingspermconcentrationinrabbitcryopreservationprotocolimplicationsforfertilitylittersizeandoffspringgrowth