Morphology, morphometry and ultrastructure of captive six-banded armadillo (Euphractus sexcinctus) sperm

Sousa, Patrícia Cunha; Santos, E.A.A.; Bezerra, José Artur Brilhante; Lima, Gabriela Liberalino; Castelo, T.S.; Fontenele‐Neto, José Domingues; Silva, Alexandre Rodrigues

doi:10.1016/j.anireprosci.2013.05.015

Cited by 14 publications

(14 citation statements)

References 22 publications

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“…The sperm structure of the head (width and length), midpiece (length) and tail (length) were measured, as well as the overall size of the cell (Sousa et al. ).…”

Section: Methodsmentioning

confidence: 99%

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Morphometry, Morphology and Ultrastructure of Ring‐tailed Coati Sperm (Nasua nasua Linnaeus, 1766)

Silva

Magalhães

Ribeiro³

et al. 2015

Reprod Domestic Animals

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The ring-tailed coati (Nasua nasua) is a procyonid whose population is in sharp decline. Therefore, studies are needed to better understand the reproduction of this animal. For this reason, this study aimed to evaluate the morphology, morphometry and sperm ultrastructure of ring-tailed coati sperm. Four captive adult males were used for this study. Slides stained with Bengal Rose were used for the morphometric and morphologic analyses. The length and width of the head were measured, as well as the length of the midpiece and tail and the total length of the sperm. Scanning electron microscopy and transmission electron microscopy were used for the ultrastructural analyses. The most obvious morphological abnormalities observed were coiled tails (6.1 ± 8.7%) and the lack of acrosomes (5.4 ± 4.4%). Regarding the morphometry, the measurements of the head (length × width), midpiece (length) and tail (length) were (mean ± SD) 6.2 ± 0.4 × 8.1 ± 0.6 μm, 14.1 ± 0.5 and 63.9 ± 4.1 μm, respectively, and the total length of the sperm was 86.1 ± 4.3 μm. Through electron microscopy, the presence of electron-lucent points in the nucleus and the presence of approximately 55 mitochondrial spirals in the midpiece were identified. The data obtained in this study provide detailed information on the sperm characteristics of coatis and may inform future research on germplasm conservation, both for this species and other threatened procyonids.

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“…The sperm structure of the head (width and length), midpiece (length) and tail (length) were measured, as well as the overall size of the cell (Sousa et al. ).…”

Section: Methodsmentioning

confidence: 99%

“…Different fields were randomly selected, evaluated by transmission electron microscopy (JEOL 1010, Japan) and photographed for later analysis and description (Sousa et al. ).…”

Section: Methodsmentioning

confidence: 99%

Morphometry, Morphology and Ultrastructure of Ring‐tailed Coati Sperm (Nasua nasua Linnaeus, 1766)

Silva

Magalhães

Ribeiro³

et al. 2015

Reprod Domestic Animals

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“…For morphometric analysis (Sousa et al . ), we pooled samples derived from both retrieval methods for each animal and then evaluated one Bengal rose smear from each animal. Images of a hundred normal sperm from random fields was captured through light microscopy (1000× of magnification) connected to image analyser software (ImageJ software, Wayne Rasband – National Institute of Health, Maryland, United States).…”

Section: Methodsmentioning

confidence: 99%

Characterization of epididymal sperm from Spix's yellow‐toothed cavies (Galea spixii Wagler, 1831) recovered by different methods

et al. 2016

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da Silva, A.M., Bezerra, J.A.B., Campos, L.B., Praxedes, E.C.G., Lima, G.L. and Silva, A.R. 2017. Characterization of epididymal sperm from Spix's yellow-toothed cavies (Galea spixii Wagler, 1831) recovered by different methods. -Acta Zoologica (Stockholm) 98: 285-291.The aim of the present study was to characterize the epididymal sperm of Spix's yellow-toothed cavy (Galea spixii) through two different recovery methods. Nine sexually mature males were euthanized and the complexes, testes-epididymis, were dissected. For each animal, one epididymis was processed by flotation method and the other was processed by retrograde flushing method, both using a TES-based buffered media. After recovery, we evaluated the sperm for motility, vigour, viability, functional membrane integrity and morphology. Morphometric data from the different sperm regions were evaluated by using an appropriate software. After recovery, both methods provide similar values for all the sperm parameters, aiming the recovery of more than 300 9 10 6 sperm, presenting >50% motile sperm, with normal morphology and functional membrane. The total sperm length in this sperm was 48.87 AE 0.1, and the sperm head presented 9.4%, on average. A notable characteristic was the prominent acrosome found in the G. spixii sperm. In conclusion, we demonstrate that either flotation or retrograde flushing methods are suitable for the recovery of sperm from cauda epididymis of Spix's yellow-toothed cavies.

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“…The sperm morphological analysis was conducted on semen smears stained with Bengal Rose, and 200 cells/slide were observed through light microscopy under magnification of 1000× (Sousa et al 2013). For ultrastructural analysis, two pools comprised of a semen sample from each animal were used; one of the pools was of fresh samples and the other, thawed.…”

Section: Methodsmentioning

confidence: 99%

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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Morphology, morphometry and ultrastructure of captive six-banded armadillo (Euphractus sexcinctus) sperm

Cited by 14 publications

References 22 publications

Morphometry, Morphology and Ultrastructure of Ring‐tailed Coati Sperm (Nasua nasua Linnaeus, 1766)

Morphometry, Morphology and Ultrastructure of Ring‐tailed Coati Sperm (Nasua nasua Linnaeus, 1766)

Characterization of epididymal sperm from Spix's yellow‐toothed cavies (Galea spixii Wagler, 1831) recovered by different methods

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

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