Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C

ContentsThe aim of this study was to evaluate powdered coconut water extender (ACP-106c; ACP Servic¸os Tecnolo´gicos Ltda, ACP Biotecnologia, Fortaleza, Ceara´, Brazil) as a diluent for freezing dog semen and the fertility after vaginal insemination of semen frozen therein. Ten ejaculates were collected from five dogs, evaluated fresh, diluted in ACP-106c, 10% egg yolk and 6% glycerol, cooled and frozen. In the first phase of the study, straws with frozen semen were thawed and immediately subjected to the same analysis as the fresh semen and, in addition, to Computer-Assisted Semen Analysis (CASA). In phase 2, 10 bitches that had been subjected to natural breeding during a preceding oestrous cycle were vaginally inseminated with thawed semen that had been re-diluted in ACP-106c. After thawing, a mean of 77% sperm motility was obtained through subjective analysis and 77.3% through CASA. Following artificial insemination, a 60% pregnancy rate was observed, resulting in a 50% parturition rate and a mean litter size of 3.4 (SEM 0.6), with 47.1% males and 52.9% females. ACP-106c can be successfully used for freezing canine semen, and vaginal deposition of such semen yields similar pregnancy rates to those reported in other studies.

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“…The diluted semen was then submitted to the freezing process previously described (Mota Filho et al. ).…”

Section: Methodsmentioning

confidence: 99%

“…The semen was diluted to achieve a final concentration of 200 9 10 6 sperm cells/ml, after the ACP-yolk-glycerol extender had been brought to a temperature of 27°C. The diluted semen was then submitted to the freezing process previously described (Mota Filho et al 2011).…”

Section: Semen Dilution and Processingmentioning

confidence: 99%

Intravaginal Artificial Insemination in Bitches Using Frozen/Thawed Semen after Dilution in Powdered Coconut Water (ACP‐106c)

Uchoa

Silva

Filho

et al. 2012

Reprod Domestic Animals

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“…A morfologia espermática foi realizada utilizando 10μL da amostra em 150μL do corante Rosa de Bengala, sendo realizado esfregaço úmido e avaliado através de microscopia óptica no aumento de 1000x, contando-se 200 células que foram classificadas como normal ou anormal (Mota Filho et al 2011).…”

Section: Methodsunclassified

“…Os parâmetros mensurados na avaliação computadorizada das amostras foram: motilidade total, motilidade progressiva, VAP, VCL, VSL, STR e LIN (Mota Filho et al 2011, Uchoa et al 2012.…”

Section: Methodsunclassified

Refrigeração do epidídimo canino a 4ºC e recuperação dos espermatozoides epididimários utilizando ACP-106c

et al. 2013

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. 2013. Objetivou-se avaliar a qualidade dos espermatozoides recuperados da cauda do epidídimo após a refrigeração do complexo testículo-epidídimo (CTE) de cães usando o diluidor ACP-106c. Foram utilizados 60 cães machos adultos, com peso de 10-20 kg.Após a eutanásia, removeu-se o CTE que foi imerso em solução fisiológica 0,9% e transportado em caixa térmica ao laboratório a 30°C. Para a refrigeração e recuperação dos espermatozoides epididimários, os 60 pares do CTE foram divididos em 4 grupos, de acordo com o tempo de refrigeração do CTE e posterior recuperação espermática: G0h, G6h, G12h e G18h, em que cada par do CTE permaneceu por zero, seis, doze ou dezoito horas a 4°C, respectivamente. A recuperação dos espermatozoides da cauda do epidídimo foi realizada pela técnica de flutuação utilizando-se o diluidor ACP-106c ou Tris. Para cada epidídimo foi adicionado 1,0 mL de um dos dois diluidores, pré-aquecidos a 37°C por 5 minutos. Em seguida foram centrifugados a 800g/5 minutos para remoção dos resíduos celulares. Avaliou-se The study aimed to assess the quality of sperm recovered from the epididymal cauda after cooling the complex testis-epididymis (CTE) of dogs using ACP-106c extender. Sixty adult male dogs, weighing 10-20 kg were used. After euthanasia, CTE was removed and it was immersed in 0.9% saline and transported to the laboratory in cooler at 30°C. For cooling and recovery of epididymal spermatozoa, the 60 pairs of CTE were divided into four groups, according to the refrigeration time of the CTE and subsequent sperm recovery: G0h, G6h, G12h and G18h, wherein each pair of CTE remained zero, six, 12 or 18 hours at 4°C respectively. The recovery of sperm epididymal tail was conducted by flotation technique using ACP-106c or Tris extender. For each epididymis, it was added 1.0 mL of either extenders, preheated to 37°C for 5 minutes. They were then centrifuged at 800g/5 minutes to remove the cell debris. Morphology, functionality and total and progressive motility, and parameters obtained by CASA were evaluated. Data were analyzed by ANOVA followed by Turkey test (P <0.05). In all parameters assessed, there was no difference between the extenders used (P>0.05). The values of total motility groups G0h, G6H, G12H, and G18h for ACP-106c were 84.4±7.7, 81.6±11.6, 88.3±6.5 and 69.5±16.9 respectively, and for Tris 85.2±8.7, 77.4±14.3, 79.0±17.8 and 65.4±17.9 respectively. A decrease in sperm quality was observed after 18 hours of cooling in both extenders. Thus it can be concluded that the ACP-106c may be used to recover the epididymal spermatozoa chilled and may be viable for up to 12h cooling.INDEX TERMS: Refrigeration, epididymis, dogs.

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“…To ensure a reliable standard, powder coconut water was developed. The use of ACP® (powdered coconut water) as a component in cryoprotectant solutions has been tested for birds (Rondon et al ) and mammals (Mota‐Filho et al ; Silva et al ). ACP‐104 is a product that was developed specifically for teleost fish (Viveiros et al ).…”

mentioning

confidence: 99%

Use of ACP‐104 at Different Dilutions for Sperm Cryopreservation of Dourado, Salminus brasiliensis

Zanandrea¹,

Weingartner²,

Zaniboni‐Filho³

2014

J World Aquaculture Soc

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The dourado, Salminus brasiliensis, is an important fish in South American river basins. The fish‐farming potential and concerns for its conservation in watersheds has stimulated studies with this species. This study compared the effect of different dilutions of two cryoprotectant solutions on the quality of cryopreserved dourado sperm (duration of motility/motility rate) post‐thawing using three different activating solutions. The cryoprotectant solution ACP‐104 was compared with the cryoprotectant solution most commonly used for dourado, which consists of a mixture of dimethyl sulfoxide, glucose, egg yolk, and distilled water. The semen was mixed following dilutions of semen : cryoprotectant solution: 1:3, 1:9, 1:15, 1:21, and 1:27. The cryopreserved samples were activated using distilled water, 0.45% NaCl or 1% NaHCO3. The results showed differences in thawed semen using the different activating solutions and cryoprotectant solutions. The glucose‐based cryoprotectant not only resulted in sperm motility equal to or better than that produced by ACP‐104, but the quality was not affected by the dilution. However, the sperm quality improved with increasing dilutions for ACP‐104. Under the conditions tested, the standard cryoprotectant solution is recommended for the cryopreservation of dourado semen because it requires a lower solution volume and, therefore, reduced storage space for the same volume of semen.

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Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C

Cited by 14 publications

References 20 publications

Intravaginal Artificial Insemination in Bitches Using Frozen/Thawed Semen after Dilution in Powdered Coconut Water (ACP‐106c)

Intravaginal Artificial Insemination in Bitches Using Frozen/Thawed Semen after Dilution in Powdered Coconut Water (ACP‐106c)

Refrigeração do epidídimo canino a 4ºC e recuperação dos espermatozoides epididimários utilizando ACP-106c

Use of ACP‐104 at Different Dilutions for Sperm Cryopreservation of Dourado, Salminus brasiliensis

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