Cryopreservation of tambaqui semen using a dry shipper and a programmed freezing machine

Abstract

Tambaqui (Colossoma macropomum) is a native freshwater fish that is of great importance for Brazilian aquaculture. Because of this importance, several techniques have been developed to improve the reproduction of this species in captivity. One of these techniques is the cryopreservation of sperm. In an effort to increase the efficiency of cryopreservation protocols, researchers have tried to determine suitable diluting solutions and freezing methods, which will provide a better post-thaw sperm quality. Thus, this study aimed to evaluate the efficiency of different diluents and freezing methods for the cryopreservation of tambaqui (C. macropomum) sperm. Samples of fresh semen were diluted in different treatments (Glucose 5% + 10% Dimethyl sulfoxide -DMSO, Glucose 5% + 10% Methyl glycol -MG, BTS + 10% DMSO and BTS + 10% MG) at a 1:9 dilution rate and frozen in a programmed freezing machine and a dry shipper. The semen samples were thawed and evaluated for vitality, sperm morphology and kinetics. Cryopreserved semen with DMSO and using the programmed freezing machine provided a greater percentage of motile sperm (15.44 ± 1.04%) after thawing compared to the dry shipper (3.99 ± 0.55%), regardless of the diluent. Additionally, DMSO showed better sperm velocities than MG regardless of the freezing method and the extender employed. A higher percentage of living spermatozoa was observed when glucose (37.28 ± 1.32%) (regardless of the freezing method and cryoprotectant) and DMSO (37.98 ± 1.25%) was used in the programmed freezing machine. For morphology, a greater amount of normal spermatozoa (46.10 ± 1.82%) was observed when the semen was cryopreserved using a freezing machine programmed with DMSO as the cryoprotectant and Glucose or BTS (38.16 ± 1.9% and 39.26 ± 1.87%, respectively) as extenders. Therefore, we suggest the use of the DMSO (10%) cryoprotectant in association with the Glucose (5%) extended in the programmed freezing machine for cryopreservation of C. macropomum semen. ResumoO tambaqui (Colossoma macropomum) é uma espécie nativa de peixe de água doce de grande importância para aquicultura brasileira. Devido a isso, diversas técnicas têm sido desenvolvidas para aperfeiçoar a reprodução desta espécie em cativeiro, dentre elas a criopreservação de sêmen de peixe. Como uma forma de melhorar os protocolos de criopreservação, tem-se buscado utilizar soluções diluidoras e métodos de congelação adequados, proporcionando uma boa qualidade seminal pós-descongelação. Dessa forma, este estudo objetivou avaliar a eficiência de diferentes diluidores e métodos de congelação na criopreservação do sêmen de tambaqui (C. macropomum). As amostras de sêmen fresco foram diluídas em diferentes tratamentos (Glicose 5% + 10% Dimetilsufóxido -DMSO; Glicose 5% + 10% Metil glicol -MG; Beltsville Thawing Solution -BTS + 10% DMSO e BTS + 10% MG) na proporção 1:9 e congeladas em máquina de congelação programada e em Dry shipper. As amostras seminais foram descongeladas e avaliadas para vitalidade, morfologia e cinética espermát...

Ions and osmolality on post‐thaw sperm motility activation of the endangered Brycon insignis (Characiformes)

“…The use of activating solutions containing Na + ion to trigger fish sperm motility has been reported for many species. The NaCl solution has been used to trigger sperm motility of fish from different orders, such as characiforms B. insignis (present study; Viveiros et al, ), P. lineatus (Viveiros, Orfão, Maria, & Allaman, ) and Colossoma macropomum (Oliveira et al, ), cypriniforms Barbus barbus (Alavi, Rodina, Policar, & Linhart, ), Puntius conchonius (Hu, Zhang, Zhou, & Zhang, ), Vimba vimba (Alavi, Kozak, Hatef, Hamackova, & Linhart, ), Clinostomus elongatus (Butts, Alavi, Mokdad, & Pitcher, ) and Cyprinus carpio (Khara et al, ), and acipenseriforms Acipenser ruthenus (Alavi, Gela, Rodina, & Linhart, ), just to name a few. The NaHCO 3 solution has been used to trigger sperm motility of characiforms species, such as B. insignis (Viveiros et al, ), Leporinus obtusidens (Taitson, Chami, & Godinho, ) and Salminus brasiliensis (Weingartner, Zanandrea, & Zaniboni, ).…”

“…Colossoma macropomum (Oliveira et al, 2016), cypriniforms Barbus barbus (Alavi, Rodina, Policar, & Linhart, 2009), Puntius conchonius (Hu, Zhang, Zhou, & Zhang, 2009), Vimba vimba (Alavi, Kozak, Hatef, Hamackova, & Linhart, 2010), Clinostomus elongatus (Butts, Alavi, Mokdad, & Pitcher, 2013) and Cyprinus carpio (Khara et al, 2014), and acipenseriforms Acipenser ruthenus (Alavi, Gela, Rodina, & Linhart, 2011), just to name a few. The NaHCO 3 solution has been used to trigger sperm motility of characiforms species, such as B. insignis (Viveiros et al, 2011), Leporinus obtusidens (Taitson, Chami, & Godinho, 2008) and Salminus brasiliensis (Weingartner, Zanandrea, & Zaniboni, 2015).…”

Development of cryopreservation methods for silver barb ( Barbodes gonionotus , Bleeker) spermatozoa by manipulation of cooling rates in dry shipper

“…Regulation of freezing rates within a dry shipper for cryopreservation of fish semen would generate a practical and field‐friendly technique to any species. Dry shipper is known to have more consistent freezing rates than other simple freezing methods, based on freezing samples in the middle of the interior space of dry shipper (Chew & Zulkafli, ; Horokhovatskyi et al, ; Koh et al, ; Oliveira et al, ). Relatively high freezing rates observed during the first 10 s of monitoring in this study were similar to the study reported by Taitson et al () who froze neotropical fish sperm ( Leporinus obtusidens ) in the dry shipper (model CP65, Taylor‐Wharton) with freezing rates of 25.7–30.8°C/s.…”

“…Dry shipper has recently been used as a freezing apparatus for the development of fish sperm cryopreservation program for aquaculture and conservation (Chew & Zulkafli, ; Gallego et al, ; Koh et al, ; Oliveira et al, ; Velasco‐Santamaría et al, ). To our knowledge, this study is the first report describing the successful cryopreservation of silver barb sperm by means of the dry shipper.…”

“…The development of efficient and convenient cryopreservation method of fish sperm is advantageous for the aquaculture industry. Recently, dry shippers have been employed successfully and cost‐effectively in freezing of fish sperm (Koh et al, ; Oliveira et al, ; Taitson, Chami, & Godinho, ). However, most studies developed the freezing methods based on exposing fish sperm samples to relatively low temperatures of the vapour phase within the dry shipper at various exposure time ranging from 10 min to 48 hr prior to plunging in liquid nitrogen container (Chiacchio, Almeida, Leal, & Viveiros, ; Horokhovatskyi et al, ; Taitson et al, ).…”