Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome
In remote alpaca breeding regions, access to advanced sperm analysis laboratories is limited. This study validates a practical cytometric method for evaluating sperm viability and acrosomal integrity in epididymal alpaca sperm using early fluorochrome staining, formaldehyde fixation, and intermediat...
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MDPI AG
2025-08-01
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| Online Access: | https://www.mdpi.com/2076-2615/15/15/2282 |
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| author | Alexei Santiani Miguel Cucho Josselyn Delgado Javier Juárez Luis Ruiz Shirley Evangelista-Vargas |
| author_facet | Alexei Santiani Miguel Cucho Josselyn Delgado Javier Juárez Luis Ruiz Shirley Evangelista-Vargas |
| author_sort | Alexei Santiani |
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| description | In remote alpaca breeding regions, access to advanced sperm analysis laboratories is limited. This study validates a practical cytometric method for evaluating sperm viability and acrosomal integrity in epididymal alpaca sperm using early fluorochrome staining, formaldehyde fixation, and intermediate storage. Thirty-two testes were transported at 5 °C, and spermatozoa were collected from the cauda epididymis. After morphometric screening, 26 samples were included. Aliquots were stained with Zombie Green (viability) and FITC–PSA (acrosomal integrity), at time zero. Each aliquot was divided for cytometric analysis at T0 (immediately), T24 (24 h after formaldehyde fixation) and T1w (1 week post-fixation). Fixed samples showed higher viability and acrosomal integrity values (T24: 70.75%, 97.24%; T1w: 71.80%, 97.21%) than T0 (67.63%, 95.89%). This may reflect fluorescence alterations associated with fixation. Strong correlations and Bland–Altman analysis confirmed consistency across time points. This method enables accurate sperm quality evaluation up to one week after collection, offering a useful tool for reproductive monitoring in field conditions without immediate analysis. Further research on ejaculated semen and field protocols is recommended. |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-08-01 |
| publisher | MDPI AG |
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| series | Animals |
| spelling | doaj-art-8396f87f1e9f48a5b3e00d4d000c933e2025-08-20T04:00:49ZengMDPI AGAnimals2076-26152025-08-011515228210.3390/ani15152282Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using FluorochromeAlexei Santiani0Miguel Cucho1Josselyn Delgado2Javier Juárez3Luis Ruiz4Shirley Evangelista-Vargas5Laboratory of Animal Reproduction, Faculty of Veterinary Medicine, Universidad Nacional Mayor de San Marcos (UNMSM), 2800 Circunvalación Av., San Borja, Lima 15021, PeruLaboratory of Animal Reproduction, Faculty of Veterinary Medicine, Universidad Nacional Mayor de San Marcos (UNMSM), 2800 Circunvalación Av., San Borja, Lima 15021, PeruLaboratory of Animal Reproduction, Faculty of Veterinary Medicine, Universidad Nacional Mayor de San Marcos (UNMSM), 2800 Circunvalación Av., San Borja, Lima 15021, PeruLaboratory of Animal Reproduction, Faculty of Veterinary Medicine, Universidad Nacional Mayor de San Marcos (UNMSM), 2800 Circunvalación Av., San Borja, Lima 15021, PeruLaboratory of Animal Reproduction, Faculty of Veterinary Medicine, Universidad Nacional Mayor de San Marcos (UNMSM), 2800 Circunvalación Av., San Borja, Lima 15021, PeruGrupo de Investigación en Biotecnología Reproductiva Animal, Dirección General de Investigación, Desarrollo e Innovación, Universidad Científica del Sur (UCSUR), Panamericana Sur Km. 19, Villa el Salvador, Lima 15018, PeruIn remote alpaca breeding regions, access to advanced sperm analysis laboratories is limited. This study validates a practical cytometric method for evaluating sperm viability and acrosomal integrity in epididymal alpaca sperm using early fluorochrome staining, formaldehyde fixation, and intermediate storage. Thirty-two testes were transported at 5 °C, and spermatozoa were collected from the cauda epididymis. After morphometric screening, 26 samples were included. Aliquots were stained with Zombie Green (viability) and FITC–PSA (acrosomal integrity), at time zero. Each aliquot was divided for cytometric analysis at T0 (immediately), T24 (24 h after formaldehyde fixation) and T1w (1 week post-fixation). Fixed samples showed higher viability and acrosomal integrity values (T24: 70.75%, 97.24%; T1w: 71.80%, 97.21%) than T0 (67.63%, 95.89%). This may reflect fluorescence alterations associated with fixation. Strong correlations and Bland–Altman analysis confirmed consistency across time points. This method enables accurate sperm quality evaluation up to one week after collection, offering a useful tool for reproductive monitoring in field conditions without immediate analysis. Further research on ejaculated semen and field protocols is recommended.https://www.mdpi.com/2076-2615/15/15/2282alpacaspermatozoafield-based evaluationflow cytometryviabilityacrosomal integrity |
| spellingShingle | Alexei Santiani Miguel Cucho Josselyn Delgado Javier Juárez Luis Ruiz Shirley Evangelista-Vargas Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome Animals alpaca spermatozoa field-based evaluation flow cytometry viability acrosomal integrity |
| title | Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome |
| title_full | Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome |
| title_fullStr | Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome |
| title_full_unstemmed | Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome |
| title_short | Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome |
| title_sort | field compatible cytometric assessment of epididymal alpaca sperm viability and acrosomal integrity using fluorochrome |
| topic | alpaca spermatozoa field-based evaluation flow cytometry viability acrosomal integrity |
| url | https://www.mdpi.com/2076-2615/15/15/2282 |
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