Trehalose in extenders for cryopreservation of Tambaqui (Colossoma macropomum) sperm.

BACKGROUND: Sugars may act as either energy substrates or non-penetrating cryoprotectants. OBJECTIVE: Inclusion of non-penetrating trehalose was tested in extenders for the cryopreservation of Tambaqui (Colossoma macropomum) sperm. MATERIALS AND METHODS: Sperm was extended 1/9 (v/v) in Beltsville Thawing Solution (BTS) with 10% DMSO (control) or 50, 100, 150 and 200 mM trehalose without 10% DMSO. Post-thawed sperm quality was evaluated, including fertilization and hatching rates, sperm motility, motility period and viability, integrity of sperm membrane and DNA, and mitochondrial functionality. RESULTS: Extenders with 100 - 150 mM trehalose achieved fertilization and hatching rates similar to those of the 10% DMSO-treated sperm samples. Trehalose at 100 and 150 mM provides better protection than 10% DMSO treatment for sperm motility, viability, DNA integrity and mitochondrial functionality. Fertilization and hatching rates were highly correlated (r = 0.95, P < 0.001). CONCLUSION: The addition of 100 - 150 mM trehalose in extender can replace 10% DMSO for the cryopreservation of C. macropomum sperm. doi.org/10.54680/fr22510110312.

GERMPLASM BANKING AND ITS ROLE IN THE DEVELOPMENT OF THE FISH GENETIC IMPROVEMENT PROGRAMME IN BRAZIL.

BACKGROUND: Over the last ten years, Brazilian fish farming has become more focused, resulting in the development of genetic improvement programmes (GIP) for two South American species Colossoma macropomum (tambaqui) and Pseudoplatystoma reticulatum (cachara). OBJECTIVE: To describe the action plan used for setting up the GIP and to detail the germplasm bank composition. MATERIALS AND METHODS: Semen of both species was collected, frozen and transported between locations in Brazil. To start the programme, full and half-sib families of both species were established from 120 males and 60 females. RESULTS: New species-specific protocols for semen cryopreservation s were established of value to commercial application in fish farming. CONCLUSION: Germplasm banking has enabled the exchange of biological material and reduced the overall GIP costs. Germplasm banking can be very important to the dissemination of the selected genetic material of these species among fish farmers.

Sperm quality of the Amazon catfish Leiarius marmoratus (Gill, 1870) after cold storage.

This study aimed at assessing the sperm quality of the Amazon catfish, Leiarius marmoratus ¸ after refrigeration without extenders. After capturing the animals and stripping of semen, the following parameters were analyzed: progressive motility, motility quality score, duration of motility and sperm morphology. An aliquot of fresh semen from each male was kept at room temperature (28 ± 2°C) as a control, for further comparison with cooled semen. The semen from each animal was stored in extenders-free individual syringes. The syringes were kept in ice within polystyrene boxes at 13 ± 2°C. For both fresh and cooled semen, seminal parameters were evaluated every one-hour interval, reaching seven hours of analysis. Fresh semen showed a significant decrease in motility, motility quality score and duration of motility remaining viable only for three hours. Progressive motility of the cooled semen displayed a negative linear pattern (P<0.05). The duration of motility increased (P<0.05), reaching its peak after three hours of storage. The motility quality score showed a quadratic pattern. No statistical differences were observed when sperm morphology was assessed (P>0.05), even though the mean values of total abnormalities have increased over the storage time. Further studies focusing on the application of this technique should be performed, including the addition of extenders and cryoprotectants for preservation of the sperm over longer periods.

Increasing storage capability of pacu (Piaractus mesopotamicus) embryos by chilling: development of a useful methodology for hatcheries management.

Cryopreservation of fish gametes has been studied extensively in the last few decades, but the successful cryopreservation of fish embryos remains elusive. However, recent studies using short-term chilling techniques have shown that it is possible to store embryos at low temperatures with no significant loss in viability. Information on cryopreservation of Neotropical freshwater fish embryos has so far been very limited in the literature. In the present study, chilling protocols for storage of pacu embryos at -8°C for up to 24 h were studied using different concentrations of sucrose in methanol. Embryos tolerated the subzero temperature for up to 6 h with no adverse effects (P > 0.05). After 12 h chilling, hatching rate of 64.0 +/- 3.5 percent was recorded. Low temperature storage of pacu embryos by chilling is detailed here for the first time. Further studies are needed to extend the storage time and to improve the hatching rate.

Use of amides as cryoprotectants in extenders for frozen sperm of tambaqui, Colossoma macropomum.

Amides were tested as cryoprotectants in comparison with glycerol and DMSO (more traditional cryoprotectants) for recovery of Colossoma macropomum (tambaqui fish) sperm. Milt was extended in Beltsville Thawing Solution, then frozen with the addition of 2%, 5%, 8%, or 11% of: (1) dimethylacetamide (DMA), (2) dimethylformamide (DMF), (3) methylformamide (MF), or with 5% glycerol or 10% dimethylsulfoxide. Fertilization rates were greatest (P<0.001) with amides; 8% DMF (91.6±1.3%), 5% DMF (88.9±1.6%), and 8% MF (83.0±1.6%), which did not significantly differ among themselves, when compared with glycerol (51.6±2.4%) and DMSO (61.9±3.1%). The best hatching rates (P<0.001) also occurred for 5% or 8% DMF and 8% MF (79.1±3.1, 87.6±1.5, and 74.8±3.0, respectively) and were also similar (P>0.05). For such treatments, both fertilization and hatching rates were similar (P>0.05) to those with fresh sperm (91.7±1.4 and 87.4±1.4, respectively). The best sperm motility across extenders (at least 55.7%) was with 5%, 8%, and 11% DMF (P<0.001). Those same treatments, along with 11% MF, provided the longest (P<0.001) period of motility (at least 1 min). The greatest sperm integrity (more than 54%) was with 5% and 11% MF and with DMA and DMF at all tested concentrations (P<0.001). The greatest (P<0.001) sperm viability (at least 31%) was for 5%, 8%, and 11% DMA, and with 8% and 11% MF, and also for DMF at all tested concentrations. Sperm DNA integrity was best (more than 50%) for 2%, 5%, and 8% MF and for DMA and DMF at all concentrations (P<0.001), whereas 2% DMA, 11% MF, 11% DMF, and the three amides at both 5% and 8% yielded the highest mitochondrial functionality (at least 44%; P<0.001); thus, 8% MF and both 5% and 8% DMF were the cryoprotectants with the best postthaw quality for C. macropomum sperm.

Qualitative parameters of the piapara semen (Leporinus elongatus Valenciennes, 1850).

Qualitative parameters of piapara semen (Leporinus elongatus) were evaluated before and after hormonal induction with carp pituitary extract at 2.5 mg.kg(-1) of live weight. The progressive motility, the spermatic vigor and the lifetime of the spermatozoa were higher before the hormonal induction (P > 0.05). The percentage of normal spermatozoa and spermatozoa with secondary pathologies did not differ (P > 0.05) between treatments: before induction (44.0 and 44.4%, respectively) and after-induction (44.3 and 46.7%, respectively). However, the percentage of primary pathologies was higher (P < 0.05) for the semen collected before induction than for the semen collected after induction; the estimates were 12.2 and 8.0%, respectively. The most frequent pathologies were the taillessness with the frequencies of 27.4 and 36.3% followed by the headlessness for which the estimates were 10.1 and 3.9%, before and after induction respectively. The semen collected before the hormonal induction presented better qualitative parameters.