In the hatchery-bred tambaqui Colossoma macropomum, spontaneous semen release does not occur, and hand-stripping produces reduced semen volume. The goal of this work is to evaluate the effects of hormonal induction with carp pituitary extract (CPE) on both qualitative (visual aspect, pH, motility, viability and morphological abnormalities) and quantitative (volume, concentration and number of spermatozoa per ejaculate) traits of tambaqui semen. Eleven males were treated with CPE (induced), and 11 were left untreated as a control (non-induced). All analysed parameters except motility and percentage of viable spermatozoa presented significant differences (p < 0.05) between the induced and non-induced treatments. CPE induction resulted in a 25-fold increase in semen volume and a 10-fold increase in the number of spermatozoa collected. However, both sperm concentration and the frequency of sperm with morphological abnormalities (commonly detached heads or bent tails) were significantly lower in CPE-induced fish. The hormonal induction of tambaqui males with CPE is efficient and positively influences some qualitative and quantitative properties of semen. Additionally, semen collection via gentle abdominal massage occurs more readily in CPE-induced fish.

Seminal features of tambaqui were evaluated after hormonal induction of spermiation with common carp pituitary extract. Seventeen adult (6.1 ± 0.9 kg, 62 ± 6 cm) males were collected from earthen ponds and transported to indoor concrete tanks. Semen was evaluated according to volume, pH, osmolality, motility, concentration, viability, sperm morphometry and morphological abnormalities. The semen of tambaqui was white and milky. The volume was 10.2 ± 5.1 mL at pH 8.0 ± 0.1, and yielded a concentration of 9.1 · 10 9 spermatozoa mL )1 while seminal plasma osmolality was 260 ± 7.3 mOsm kg )1 . The percentage of viable sperm cells was determined with 97.0 ± 2.0% and 87.0 ± 8.0% using an eosin-nigrosin staining and fluorescent live-dead kit (propidium iodide and SYBR-14), respectively. A negative correlation ()0.65) between semen pH and sperm motility was observed suggesting this feature might influence the tambaqui sperm kinetics. Morphometrically the spermatozoa were on average 35.48 ± 1.55 lm long with a roundish head (mean length: 2.73 ± 0.21 lm; mean width: 2.58 ± 0.18 lm; n = 250 spermatozoa), without an acrosome, and presented a long midpiece (2.90 ± 0.52 lm) and flagellum (29.84 ± 1.63 lm). About 15.8% of the spermatozoa carried morphological abnormalities, with bent tail being the most frequent defect (7.81 ± 3.12%). The characterization of tambaqui semen favors the development of more precise and efficient procedures for its analysis and utilization in controlled breeding.

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Two new species, Hyphessobrycon frickei Guimarães, Brito, Bragança, Katz & Ottoni sp. nov. and H. geryi Guimarães, Brito, Bragança, Katz & Ottoni sp. nov., are herein described, based on seven different and independent species delimitation methods, and on molecular and morphological characters, making the hypothesis of these new species supported from an integrative taxonomy perspective. They belong to the “Rosy tetra” clade, which is mainly characterized by the presence of a dark brown or black blotch on the dorsal fin and the absence of a midlateral stripe on the body. These two new species are distinguished from the other members of this clade mainly by the arrangement, shape and color pattern of humeral and dorsal-fin spots, as well as by other characters related to scale counts and body pigmentation. The placement of the new species within the “Rosy tetra” clade was based on the combination of morphological character states mentioned above and corroborated by a molecular phylogenetic analysis using the mitochondrial gene cytochrome oxidase subunit 1. In addition, a new clade (here termed Hyphessobrycon copelandi clade) within the “Rosy tetra” clade is proposed based on molecular data, comprising H. copelandi, H. frickei sp. nov., H. geryi sp. nov. and a still undescribed species. Our results corroborate the occurrence of hidden species within the “Rosy tetra” clade, as suggested by previous studies.