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2 results on '"Trypan blue/Giemsa"'

1. Evaluation of buffalo semen by Trypan blue/Giemsa staining and related fertilityin vitro

Author

Bianca Gasparrini, E. Mariotti, L. Attanasio, A. De Rosa, L. Boccia, R. Di Palo, Boccia, Lucia, DI PALO, Rossella, DE ROSA, Anna, Attanasio, Laura, Mariotti, E., and Gasparrini, Bianca

Subjects
endocrine system, medicine.medical_specialty, 040301 veterinary sciences, animal diseases, media_common.quotation_subject, Semen, Fertility, Biology, Giemsa stain, 0403 veterinary science, Andrology, chemistry.chemical_compound, fluids and secretions, medicine, Trypan blue/Giemsa, Blastocyst, reproductive and urinary physiology, lcsh:SF1-1100, media_common, buffalo, Gynecology, urogenital system, 0402 animal and dairy science, semen, 04 agricultural and veterinary sciences, 040201 dairy & animal science, Sperm, Semen, Buffalo, Trypan blue/giemsa, Quality evaluation, quality evaluation, In vitro, medicine.anatomical_structure, Double staining, chemistry, Animal Science and Zoology, Trypan blue, lcsh:Animal culture
Abstract

The aim of this work was to verify the feasibility of an easy, quick double staining technique for evaluation of frozen-thawed semen to predict the fertilizing capability in vitro of buffalo bulls. In Experiment 1, frozen-thawed semen from 6 bulls was stained with double Trypan blue/ Giemsa and the incidence of acrosome-intact live (AIL), acrosome-intact dead (AID), acrosome-lost live (ALL) and acrosome-lost dead (ALD) sperm was recorded. In Experiment 2, sperm from the same bulls were used to fertilize in vitro matured oocytes. The data obtained confirm that there is a strong “bull effect” in buffalo species, with differences in the percentage of AIL sperm at thawing, in cleavage and blastocyst rates among bulls. Interestingly, it was found that this staining technique can be used for a preliminary screening to select semen to use for IVF, as shown by the correlation existent between the percentages of acrosome-intact viable sperm cells at thawing and the blastocyst yields for 4/6 bulls.

Published
2007
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2. Evaluation of In Vitro Capacitation of Buffalo Frozen/Thawed Sperm by Different Techniques

Author

Elkhawagah, A. R., Longobardi, V., Gasparrini, B., Sosa, G. A., Bifulco, G., Abouelroos, M.E. A., el-ghafar, A.E. Abd, Camapnile, G., Elkhawagah, A. R., Longobardi, V., Gasparrini, B., Sosa, G. A., Bifulco, G., Abouelroos, M.E. A., el-ghafar, A.E. Abd, and Camapnile, G.

Abstract

This study aimed to determine the most reliable method to evaluate capacitation of buffalo frozen/thawed sperm. Frozen/thawed sperm cells were incubated in Tyrode albumin lactate pyruvate medium (TALP) in absence of capacitating agents (control) and in presence of 10 µg/ml heparin for 2 and 4 h. Capacitation was assessed by Trypan blue/Giemsa after lysophosphatidylcholine (LPC) exposure, chlortetracycline (CTC) fluorescence assay and immune-localization of tyrosine phosphorylated protein. Furthermore, we evaluated the effect of heparin on penetration, cleavage rates and kinetics of embryo development after heterologous IVF. The percentage of LPC-induced acrosome reacted (AR)-sperm increased (P<0.05) with heparin compared to the control after 2 h (28.2 vs 24.4%, respectively) and 4 h (35.1 vs 32.0 %, respectively). No differences in CTC pattern B (capacitated sperm) were found between groups and incubation times (on average 63%). On the contrary, heparin decreased (P<0.01) the percentage of tyrosine phosphorylation pattern A after 2 and 4 h (34.3 and 35.3%, respectively) compared to the control (54.5 and 51.8%, respectively) and increased (P<0.01) that of pattern EA after 2 and 4 h (59.2 and 54.2 %, respectively) compared to the control group (44.7 and 45.2 %, respectively). Both cleavage and penetration rates, as well as the percentage of fast developing embryos, were higher (P<0.01) in the heparin-treated group (77.2, 80.4 and 74.0 %, respectively) compared to the control (56.6, 58.0 and 55.2 %, respectively). In conclusion, Trypan blue/Giemsa staining to evaluate LPC-induced AR and tyrosine protein phosphorylation assay can be successfully used to evaluate capacitation of buffalo frozen/thawed semen.

Published
2014

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