Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders

Lima, David Baruc Cruvinel; Silva, T.F.P.; Cortez, Annira Aquino; Pinto, José Nicodemos; Magalhães, Francisco Felipe de; Caldini, B.N.; Silva, L.D.M.

doi:10.1590/1678-4162-8653

Cited by 10 publications

(11 citation statements)

References 18 publications

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“…Estudos mostram que o uso do ACP como diluente seminal é uma alternativa promissora e eficaz na conservação de importantes características espermáticas em várias espécies animais [2,9,14,18,22]. Atualmente, poucos são os estudos sobre resfriamento seminal de peixes que envolvem o uso do ACP-104 como diluente, existindo apenas estudos com Prochilodus lineatus [22] e Colossoma macropomum [15], sendo os resultados de cinética semelhantes aos encontrados na presente pesquisa.…”

Section: Discussionunclassified

“…Atribui-se o sucesso do ACP como diluente seminal à sua composição, com poucos fosfolipídeos e rica em proteínas, sais, açúcares, vitaminas e fatores de crescimento, como o ácido indol-3-acético (AIA) [2]. Estudos demonstraram que o AIA estimula e mantém uma maior motilidade para sêmen de caprinos e ovinos [14], além de proporcionar um maior número de espermatozoides com membrana íntegra [9] e com morfologia normal [2].…”

Section: Discussionunclassified

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Influence of Cooling Time and Diluents on the Freezability of Prochilodus brevis semen

Nascimento

Leite-Castro

Montenegro

et al. 2017

Acta Scientiae. Vet.

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Background: Prochilodus brevis is a rheophilic fish of economic and ecological importance. However, anthropic action has made its population vulnerable. Thus, the development of reproductive biotechnologies, such as seminal conservation, is necessary to subsidize their fish farming. However, seminal collections are often performed in places with few laboratory resources, demanding studies to determine the maximum time for which sperm can be cooled, as well as its process until frozen. Thus, the present study aimed to evaluate the influence of cooling time and the presence of dilution solutions on cryopreservation of P. brevis semen.Material, Methods & Results: After seminal collection, nine pools were formed and analyzed for seminal pH, concentration, membrane integrity, morphology and spermatic kinetics - motility, curvilinear velocity (VCL), average path velocity (VAP) and straight line velocity (VSL). After the analysis of the pools in natura (control 1), they were processed as follows: 1)- immediate freezing (control 2); 2)- cooling: undiluted, diluted in coconut water powder (ACP-104) or diluted in 5% glucose, followed by cooling at different times (6, 12, 24 or 48 h); 3)- Post-refrigeration freezing: the pools were diluted in their respective diluents and 10% dimethyl sulfoxide. After 15 days, the samples were thawed and analyzed for the aforementioned parameters. For the cooled and post-thawed semen, a completely randomized design with 2 (diluent × cooling time) and 3 (storage form × cooling time and storage form × diluent) factors, respectively, was utilized. ANOVA and Dunnett tests were applied to compare the means. In case of seminal cooling, there was no difference (P > 0.05) in sperm motility between control 1 and the undiluted and diluted treatments in ACP-104 for up to 24 h. After 48 h, only the VCL of the sample diluted in ACP-104 was similar (P > 0.05) to that of control 1. When comparing forms of storage (undiluted, diluted in ACP-104 or diluted in glucose) and cooling times, the undiluted samples and the samples diluted in ACP-104 were better (P < 0.05) for all the kinetics parameters analyzed, than those diluted in glucose after 24 h. After 48 h, the cooled semen diluted in ACP-104 presented greater (P < 0.05) motility than the other treated semen samples. The samples diluted in glucose for 48 h presented lower spermatic velocity (P < 0.05) than those subjected to other treatments. Regardless of the diluent used, the post-thawed semen and the cooled semen diluted for 6 h, presented higher sperm kinetic values (P < 0.05) than those of control 2 and other treated samples. Overall, the samples diluted in ACP-104 showed satisfactory results when cooled for up to 48 h or cooled for up to 6 h and frozen.Discussion: This is the first study that froze semen from P. brevis after cooling. Although glucose is a commonly used diluent during seminal freezing and has good post-thawing stability for this species, it is not recommended for cooling before seminal freezing, as prolonged exposure of spermatozoa to glucose may cause osmotic stress to sperm cells. Conversely, good results with ACP-104 might be because of its rich composition, mainly the presence of indole-3-acetic acid (IAA), an auxin with proven potential for seminal conservation of other species. Therefore, for fertilization trials, it is recommended to use ACP-104 as diluent for seminal cooling of P. brevis for up to 48 h or semen that has been frozen after cooling in ACP-104 for a maximum of 6 h.

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Section: Discussionunclassified

Influence of Cooling Time and Diluents on the Freezability of Prochilodus brevis semen

Nascimento

Leite-Castro

Montenegro

et al. 2017

Acta Scientiae. Vet.

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“…The recovered epididymal was transferred to a 1.5‐ml conical tube, preheated at 37°C. Each sample was previously evaluated for total motility and progressive motility by computer system analysis (CASA), using the Sperm Class Analyser ® software (SCA Microptic SL, version 5.3.0.1), previously configured for the species (Filliers et al,; Lima et al, ).…”

Section: Methodsmentioning

confidence: 99%

Morphological and morphometric characterization of domestic cat epididymal sperm

Barbosa

Silva

Tabosa

et al. 2019

Reprod Domestic Animals

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Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm2, while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.

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“…In prepubertal animals, different techniques may be used to obtain sperm, as semen collection with an artificial vagina (Lambo et al., ), electroejaculation (Henry et al., ) or urethral catheterization (Lueders, Luther, Scheepers, & Horst, ), as well as the recovery of epididymal spermatozoa (Lima et al., ). On the other hand, cryopreservation of testicular fragments represents the only possibility to preserve the fertility of prepubertal animals.…”

Section: Introductionmentioning

confidence: 99%

Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Lima

Silva

Morais

et al. 2016

Reprod Domestic Animals

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The cryopreservation of testicular tissue is presented as the only alternative for the preservation of genetic material from prepubertal animals. However, this biotechnology is still being tested. The objective of this study was to evaluate the effect of different associations of cryoprotectants and the potential of cell proliferation after vitrification of testicular tissue of prepubertal cats. Five testicular pairs from five prepubertal cats were used, and each pair was divided into four fragments. Of these, one fragment composed of the control group (CG) and the rest were distributed in experimental groups according to the associations of cryoprotectants to be tested (dimethyl sulphoxide (DMSO)/glycerol (GLY); ethylene glycol (EG)/GLY) or DMSO/EG) in a final cryoprotectant concentration of 5.6 m. The fragments were submitted to vitrification, and after one week, fragments were heated and processed for histomorphological evaluation and quantification of nucleolar organizer regions (NORs). DMSO/GLY did not differ from CG and was superior to the other vitrified groups, as to cell separation and degree of shrinkage of the basal membrane. Concerning cell differentiation, visibility of the nucleus and nuclear condensation, all the vitrified groups were inferior to CG; however, DMSO/EG was inferior to DMSO/GLY and EG/GLY, which did not differ among themselves. CG was superior to all groups in quantification of NORs. DMSO/EG was inferior to all others, and there was no difference between DMSO/GLY and EG/GLY. The association DMSO/GLY presented the best preservation of tissue integrity and potential of cell proliferation after vitrification of the testicular tissue of prepubertal cats.

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Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders

Cited by 10 publications

References 18 publications

Influence of Cooling Time and Diluents on the Freezability of Prochilodus brevis semen

Influence of Cooling Time and Diluents on the Freezability of Prochilodus brevis semen

Morphological and morphometric characterization of domestic cat epididymal sperm

Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

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