Tomato pomace, a by-product generated during tomato processing, was collected at a large tomato processing industry. The by-product was mainly constituted of tomato skin (61.5%), and presented high moisture content (66.58 g.100g wet basis). Among the nutrients, the highest content was of dietary fibre, followed by proteins and fat (50.74, 20.91, 14.14 g.100g d.w., respectively). The pomace has high in vitro antioxidation capacity, especially when measured with the TEAC assay (224.81 μmol Trolox equivalent 100g d.w.). This is due especially to the high amount of lycopene remaining in the by-product after processing (446.9 μg.g d.w). The waste was fractioned into skin and seed fractions by sedimentation, resulting in the increase of lycopene yield by 55%, when using skin fraction as the source material in place of the whole pomace. This by-product shows great potential for being used as a source of the ingredients of high nutritional value, especially dietary fibre and lycopene.

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Currently, aquaculture has been growing due to factors such as increased fish consumption and production, population growth, higher family income and urbanization (Food and Agriculture Organization of the United Nations (FAO), 2019). Therefore, studies that focus on investigating the reproductive biology of fish have been intensified to develop species conservation and genetic improvement programmes (Denniston, Michelet, & Godke, 2000), as well as to enable an increase in aquaculture production and reduce pressure on natural stocks. Thus, species belonging to genus Prochilodus, like Prochilodus brevis, a rheophilic species native to Northeast Brazil, have shown increasing potential for use in Brazilian aquaculture (Peixe Br, 2019) to meet the great demand for fish, as they are among the most important species in both commercial and subsistence continental

Background: Seminal cryopreservation is a technique that optimizes aquacultural production, as it requires less breeding and enables reproduction outside of the breeding season. This technique also helps to preserve species, thus reducing the pressure on the natural stocks. Several studies have sought to develop freezing protocols that result in semen of a good quality. However, some studies do not evaluate the ability of frozen semen to produce viable larvae. Therefore, the aim of this study was to verify the fertilizing capacity of the frozen semen of Prochilodus brevis.Materials, Methods & Results: Semen from twenty adult males of the Brazilian bocachico was collected and evaluated to establish the total motility, curvilinear velocity, straight linear velocity, average path velocity, membrane integrity, pH, and concentration. Six pools were formed, each of which was diluted in a freezing medium containing 5% glucose with 10% dimethyl sulfoxide (DMSO) or 5% glucose with 10% methyl glycol (MG). The samples were loaded into 0.25 mL French straws, frozen in a dry shipper, and stored in a liquid nitrogen canister. The semen was then thawed and evaluated to establish the total motility, curvilinear velocity, straight linear velocity, average path velocity, and membrane integrity. For the fertilization test, four females were used. The oocytes from each female were divided into three batches and fertilized with either fresh or cryopreserved semen. The rates of fertilization, hatching, and larval survival were then measured. Data were expressed as the mean ± standard deviation and analyzed using SAS (2002). The frozen semen with glucose + DMSO was significantly higher (P < 0.001) than the frozen semen with glucose + MG, in all seminal quality parameters evaluated (63.95 ± 15.88% and 25.36 ± 3.53% for the motility, 36.38 ± 7.02 μm.s-1 and 20.45 ± 2.84 μm.s-1 for the curvilinear velocity, 19.26 ± 2.74 μm.s-1 and 3.03 ± 1.40 μm.s-1 for the straight linear velocity, 25.70 ± 6.51 μm.s-1 and 6.90 ± 1.12 μm.s-1 for the average path velocity, and 63.58 ± 6.95% and 35.58 ± 11.26% for the membrane integrity, respectively). MG were very close to zero and not statistically significant. Regarding these same parameters, there were no significant differences (P > 0.05) when the fresh semen was compared to the cryopreserved semen with glucose + DMSO (36.25 ± 2.5% and 29.16 ± 5.64% for the fertilization rate, 38.56 ± 11.23% and 29.33 ± 11.75% for the hatching rate, and 11.59 ± 5.16% and 7.63 ± 5.46% for the larval survival rate, respectively).Discussion: This is the first study of the artificial fertilization of Prochilodus brevis using cryopreserved semen. Seminal quality parameters are important for predicting the success of the cryopreservation technique, however, in vivo tests are essential to confirm such success. Thus, obtaining larvae is a major step towards the standardization of a cryopreservation protocol for a particular species. It is known that cryopreservation reduces the seminal quality but is a necessary process for the conservation of male gametes in the long term and, as shown in this study, good results can be obtained. In this study, the best results were obtained with the inclusion of DMSO in the freezing medium. This effect can be attributed to DMSO having a very low molecular weight, which decreases the formation of ice crystals. Considering the results obtained, we concluded that it is feasible to obtain larvae of the Brazilian bocachico using frozen semen in a 5% glucose solution with 10% DMSO.

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SummaryProchilodus brevis is a rheophilic species with a threatened natural population that promotes studies aimed at optimizing reproduction in captivity. The correct quantity of inseminating dose and activating solution volume significantly improves fertilization rates, thereby increasing productivity in captivity. The objective of this study was to determine the proportion of sperm per oocyte and the ideal volume of activating solution to be used in the assisted fertilization of P. brevis. Gametes were collected and fertilization performed in two steps. In step 1, the ideal proportion of spermatozoa was determined based on the fertilization rate:oocyte by testing six doses of semen: D1 = 30 × 103, D2 = 150 × 103, D3 = 300 × 103, D4 = 3 × 106, D5 = 5 × 106, and D6 = 10 × 106. In step 2, the fertilization and hatching rates were evaluated using different volumes of activating solution (V1 – 25 ml, V2 – 50 ml, V3 – 75 ml,V4 – 100 ml, V5 – 125 ml, and V6 – 150 ml). A linear regression equation was estimated from steps 1 and 2. The Student–Newman–Keuls test was used to compare the means. In step 1, the percentage of fertilization increased linearly, reaching a plateau of 51.69%. In step 2, the best fertilization rates were obtained with an estimated ideal volume of 75.64 ml per 2 ml of oocytes. Therefore, the proportion of 928,410.29 sperm:oocyte, associated with the volume of 75.64 ml of water per 2 ml of oocytes, provided the maximum reproductive performance for P. brevis.

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The study aimed to evaluate the in vitro antioxidant action of glycosaminoglycans (GAGs) from the skin of Oreochromis niloticus, and to determine their ideal concentration to supplement the sperm freezing medium of Prochilodus brevis. In experiment 1, the in vitro antioxidant properties of GAGs were verified through the analysis of DPPH, chelating ferrous ability, and total antioxidant capacity. In experiment 2, milt pools were formed, which were frozen in solution supplemented or not with different GAGs concentrations: 0 (control), 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg mL-1 (total of 10 treatments). The samples were evaluated for membrane integrity, DNA integrity, sperm morphology, and sperm kinetics. The results of experiment 1 showed that the GAGs exhibited, with the increase of the concentration, significant antioxidant action, for all the evaluated tests, mainly in the chelating ferrous ability. In experiment 2, it was observed that the increase of GAGs concentration decreased kinetic parameters (P < 0.05), however, the control and 0.5 mg mL-1 GAGs concentration showed similar results. For the other parameters (membrane integrity, DNA integrity, and sperm morphology), there was no decrease in results with the increase of GAGs concentration. In conclusion, GAGs extracted from O. niloticus skin have antioxidant action, and the concentration of 0.5 mg mL-1 was the most adequate to supplement the P. brevis sperm-freezing medium.