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4 results on '"Trypan blue/Giemsa"'

1. Effect of Osteopontin on Buffalo Sperm Capacitation.

Author

Mariotti, E., Novoa, V. Suárez, Boccia, L., Rubessa, M., Di Francesco, S., De Blasi, M., Longobardi, V., and Gasparrini, B.

Subjects
OSTEOPONTIN, SPERMATOZOA, WATER buffalo, FERTILIZATION in vitro, ACROSOME reaction, SPERM motility, HEPARIN, REPRODUCTION
Abstract

The aim of this study was to evaluate the effect of osteopontin (OPN) to induce capacitation of buffalo spermatozoa in vitro. Capacitation was assessed by the ability of sperm to undergo the acrosome reaction (AR) after incubation with lysophosphatidylcholine (LPC), a known inducer of AR only on capacitated spermatozoa. Buffalo frozen-thawed spermatozoa were incubated for 2 and 4 h in TALP medium (control) or in TALP medium supplemented with 0.01 mM heparin, or with 0.1, 1, 10 μg/mL OPN. Two-hours incubation with 1 or 10 μg/mL OPN significantly increased (P<0.05) the percentage of AR compared to the control group whereas heparin and 0.1μg/mL OPN provided intermediate results. After 4 hours incubation the same higher concentrations tested caused a further increase (P <0.01) compared with the control. Furthermore, after 4 hours the treatment with heparin enhanced capacitation compared with the control, as indicated by higher percentages of spermatozoa with AR (P <0.05). These results show the possibility to significantly improve the efficiency of capacitation in vitro in buffalo species by incubating sperm in the presence of OPN. [ABSTRACT FROM AUTHOR]

Published
2010

2. 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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3. 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

4. Evaluation of stallion semen by trypan blue /giemsa staining and related fertility

Author

SANTESE G., BOCCIA L., SPADETTA M., DE ROSA A., ATTANASIO, LAURA, GASPARRINI, BIANCA, CAMPANILE, GIUSEPPE, DI PALO, ROSSELLA, W. Martin Rosset, Santese, G., Boccia, L., Spadetta, M., Attanasio, Laura, DE ROSA, A., Gasparrini, Bianca, Campanile, Giuseppe, and DI PALO, Rossella

Subjects
fertility, stallion, Trypan blue/Giemsa
Abstract

The double staining Trypan blue/Giemsa has been used to evaluate the semen of two stallions (A and B) bred in the same farm. In order to compare the fertility rate of the last mating season with the results of this staining technique, semen samples were collected at 10 days intervals and stained according to Kovács and Foote (1992). Sperm cells were then classified as: live with intact acrosome (VCA), live with acrosome reacted (VAR), dead with intact acrosome (MCA), dead with acrosome reacted (MAR), dead with damaged acrosome (MAD), only sperm head (ST). Mating data and pregnancy rate at 40 days of gestation were collected in order to evaluate the first cycle pregnancy rate and the seasonal pregnancy rate. Differences of pregnancy rate between stallions were analyzed by Chi Square test. Differences of sperm cells percentage in the staining classes were analyzed by ANOVA, after the angular transformations. Seasonal pregnancy rate were different between stallions (85,7% vs 52,9% in A vs B; P

Published
2006

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