Viabilidade do sêmen de tatus-peba (Euphractus sexcinctus) centrifugado e diluído em Tris ou agua de coco em pó

Sousa, Patrícia da Cunha; Santos, E.A.A.; Silva, Andréia Maria; Castelo, Thibério de Souza; Peixoto, Gislayne Christianne Xavier; Freitas, Carlos Iberê Alves; Silva, Alexandre Rodrigues

doi:10.1590/0103-8478cr20131015

Cited by 3 publications

(3 citation statements)

References 16 publications

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“…On a recent study developed by our team, we demonstrated that a Tris extender is more effective in maintaining armadillo's sperm viability for 120 min at 34 o C than a coconut water extender (Sousa et al 2014). For this reason, we chose the Tris as the diluting medium for this first attempt on armadillos' sperm freezing.…”

Section: Discussionmentioning

confidence: 99%

“…In fact, an old study had yet revealed that armadillo sperm surface present some difficulties in incorporate fluorescent markers (Bedford & Millar 1978). In this sense, a diluted sample was used as the control for the experiment, once extender promoted the reduction in medium viscosity and allowed the sperm rolleaux dissolution (Sousa et al 2014), thus allowing the interaction among sperm membrane surface and fluorophores. This technique revealed that the mitochondria of the six-banded armadillo's sperm seems to be very sensible to the cryoinjuries, and this statement corroborates to the findings derived from the transmission electron microscopy.…”

Section: Discussionmentioning

confidence: 99%

“…Each sample of fresh semen was diluted with Tris [Tris-hydroxymethyl aminometane, 3.028g, citric acid monohydrate 1.78g and D-fructose 1.25g], plus 20% egg yolk (Sousa et al 2014). Samples were cooled for 40 min in a 5 L isothermic box (Isoplast ® , Fortaleza, CE, Brazil), at 15°C, and further transferred to an incubator (Quimis ® , Diadema, SP, Brazil) for 30 min to reach 4°C, being evaluated at this moment.…”

Section: Methodsmentioning

confidence: 99%

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Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

et al. 2016

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RESUMO.-[Identificação de danos ultraestruturais e funcionais em espermatozoides de tatus-peba (Euphractus sexcinctus) oriundos da criopreservação.] O objetivo deste estudo foi criopreservar o sêmen de tatus-peba (Euphractus sexcinctus) em diluente Tris-gema e glicerol, e determinar os danos causados pelo processo de congelação-descongelação, utilizando marcadores fluorescentes e análise ultraestrutural. As amostras de sêmen (n=11) coletadas de 4 tatus-peba adultos por eletroejaculação foram criopreservadas em diluente Tris acrescido de 20% de gema de ovo e 3% de glicerol, em curva rápida de congelação. A análise clássica das amostras foi realizada após a diluição, refrigeração e descongelação, seguida por análise de fluorescência, utilizando uma combinação de sondas flu- The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide -PI and Hoechst -H342), and mitochondrial activity (CMXRos -Mito Tracker Red ® ). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol. Além disso, as imagens obtidas por microscopia eletrônica de transmissão revelaram que espermatozoides congelados-descongelados apresentaram membranas plasmáticas danificadas, modificações nucleares como alterações na cromatina, e alterações acrossomais relativas à capacitação espermática. Em conclusão, este estudo é a primeira tentativa de criopreservação de sêmen em uma espécie de tatu, e nos auxiliou a identificar pontos críticos no processo de ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

et al. 2016

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Ex Situ Approaches for the Conservation of Genetic Resources in the Superorder Xenarthra

Fernandes

Praxedes

Freitas

et al. 2023

Biopreservation and Biobanking

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Monitoring the reproductive physiology of six‐banded armadillos (Euphractus sexcinctus, Linnaeus, 1758) through different techniques

Campos

Peixoto

Lima

et al. 2016

Reprod Domestic Animals

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The aim of this study was to monitor the oestrous cycle using vaginal cytology, ultrasound and measurement of hormone levels associated with the modification of external genitalia in female Euphractus sexcinctus. Five adult female six-banded armadillos were used for the study. Every three days, we chemically restrained the animals with a combined dose of ketamine and xylazine for 90 days. On each occasion, we conducted vaginal cytology and monitored the alterations in the vulval appearance. In addition, we obtained blood samples for serum estradiol and progesterone analysis and evaluated the ovaries by ultrasonography (8 MHz). As results, at least two entire cycles were monitored per female as based on external oestrous signs. We determined that six-banded armadillos' oestrous cycle lasts 23.5 ± 3.12 days, comprising 8.8 ± 1.4 days for oestrogen phase, in which we verified vaginal bloody discharge, vulvar oedema, presence of mucus and ease of introduction of the swab. During oestrus, females presented an oestrogen peak of 240.66 ± 12.69 pg ml(-1) , on average, with a positive visualization of ovary follicles by ultrasound. The progesterone phase lasts 15.62 ± 2.1 days, characterized by the absence of bloody secretion and difficulty in introducing the swab; there was verification of a progesterone plateau of 10.83 ± 1.86 ng ml(-1) , on average, with identification of corpora lutea in 60% of the ovaries. This is apparently the first description of the six-banded armadillos' oestrous cycle, which proves the efficiency of a multiparametric analysis to monitor it.

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Viabilidade do sêmen de tatus-peba (Euphractus sexcinctus) centrifugado e diluído em Tris ou agua de coco em pó

Abstract: Viabilidade do sêmen de tatus-peba (Euphractus sexcinctus) centrifugado e diluído em Tris ou agua de coco em pó.Ciência Rural, v.44, n.9, set, 2014.

Cited by 3 publications

References 16 publications

Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

Ex Situ Approaches for the Conservation of Genetic Resources in the Superorder Xenarthra

Monitoring the reproductive physiology of six‐banded armadillos (Euphractus sexcinctus, Linnaeus, 1758) through different techniques

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