Bee pollen in zebrafish diet affects intestinal microbiota composition and skin cutaneous melanoma development.

Bee pollen is recommended as dietary supplement due to immunostimulating functions including antioxidant, anti-inflammatory and anti-carcinogenic properties. Nevertheless, the effectiveness of such properties is still not well understood. As diet can be associated with animal performance, microbiota modulation and potentially factor for cancer, this study aimed to analyze if bee pollen could influence growth, gut microbial and skin cutaneous melanoma development in zebrafish. Control diets based on commercial flakes and Artemia were compared with the same diet supplemented with bee pollen. Fish weight gain, increased length, intestinal bacteria metagenomics analysis, serum amyloid A gene expression and cutaneous melanoma transplantation assays were performed. Bee pollen affected microbiota composition and melanoma development. Differential abundance revealed higher abundance in the control group for Aeromonadaceae family, Aeromonas and Pseudomonas genus, A. sobria, A. schubertii, A. jandaei and P. alcaligenes species compared with pollen diet group. Pollen group presented higher abundance for Chromobacterium genus and for Gemmobacter aquaticus, Flavobacterium succinicans and Bifidobacterium breve compared with control group. Unexpectedly, fish fed with bee pollen showed higher tumor growth rate and larger tumor size than control group. This is the first study to report intestinal microbial changes and no protective cancer properties after bee pollen administration.

Cannibalism rate and mLeptin expression are influenced by photoperiod and diets in Piracanjuba, Brycon orbignyanus (Valenciennes, 1850) larvae.

Piracanjuba (Brycon orbignyanus) is a species with great productive potential, and during its larval phase, it presents intense cannibal activity. The photoperiod and diet are primary feed behaviour and cannibalism modulators to fishes. This experiment aimed to verify the effect of different photoperiods and diets in Piracanjuba larviculture. Larvae were kept under different photoperiods - 12 h light: 12 h dark (12 L: 12D); 24 h light:00 h dark (24hL: 00D) - Larvae were fed with Artemia nauplii and a formulated micro-diet in a factorial scheme for 10 days, and at the end of the experimental period, the influences of the treatments on performance and quantitative expression of mLeptin and mBmall1 were evaluated. In order to quantify the expression of mLeptin and mBmall1, qPCR adopting ß-actin and Elongation Factor 1 as endogenous genes was used. The primers for all the analysed transcripts were obtained through multiple sequences alignments of different fish species. It was observed that the diet and photoperiod influence the performance of Piracanjuba (B. orbignyanus) larvae in the initial phase of larviculture. Feeding with artemia nauplii and the photoperiod of 24 L:00D reduce cannibalism rates in intensive Piracanjuba larviculture. The results on the rate of cannibalism, rate of survival and the relative expression of mLeptin are related to the survival rate of the larvae, and it is inversely proportional to the cannibalism rate. The expression levels of mBmall1 showed a correlation with the final weight of the larvae. Piracanjuba Larvae under a photoperiod of 24 light and fed Artemia nauplii showed more significant levels of mLeptin expression.

Effects of melatonin supplementation on the quality of cryopreserved sperm in the neotropical fish Prochilodus lineatus.

The cryopreservation process causes damage to sperm structures and supplementation of the cryoprotective medium is an alternative to reduce these damages. The aim of this study was to determine the effect of melatonin supplementation on post-thaw sperm quality in Prochilodus lineatus. The cryoprotective medium was supplemented with 2.00, 2.75, 3.50, and 4.25 mM melatonin, and the control group (without melatonin). Sperm motility parameters, membrane integrity, sperm morphology, oxidative stress (lipid peroxidation and enzymatic activity), and fertilization capacity were analyzed in the post-thaw sperm. Samples cryopreserved with 2.00 mM melatonin yielded higher sperm motility rate than other treatments with the addition of melatonin. Sperm curvilinear velocity (VCL) and average path velocity (VAP) were higher in samples containing 2.00 mM melatonin than in other treatments. Samples from control and with 2.00 mM melatonin presented higher membrane integrity and morphological normality than samples containing 4.25 and 2.75 mM melatonin, respectively. Regarding oxidative stress, lower lipid peroxidation (LPO) occurred in 2.75 and 3.50 mM melatonin, compared to control. While higher enzyme activity of catalase (CAT) occurred in the control than in other treatments, no differences were observed in the activity of superoxide dismutase (SOD). Higher fertilization and hatching rates occurred at 2.75 mM melatonin compared to 4.25 mM. Although no significant differences were observed in LPO between the control and samples supplemented with 2 mM melatonin, it was observed that this dosage of melatonin allowed higher VCL and VSL and reduced values of CAT. However, as there are no differences in motility and fertilization rates between the control and the 2 mM concentration, it is suggested that further studies be carried out with lower concentrations of melatonin in order to determine its effectiveness as an antioxidant in the sperm of this species.

Sperm characterization and cryopreservation of the endangered freshwater fish Chirostoma estor (Atheriniformes).

The knowledge of the physiology of sperm of an endangered species allows the implantation of reproductive biotechnologies that aim at conservation. The aim of this study was to characterize fresh sperm and evaluate different cryopreservation solutions for sperm in Chirostoma estor. The characterization of Chirostoma estor fresh sperm (n = 22 males) was performed through analyzes of sperm concentration, membrane integrity, sperm morphology, motility rate, motility quality score, and motility duration. For cryopreservation (n = 42 males), 3 extenders (BTS™, MIII™, or Androstar Plus™) in combination with 2 permeable cryoprotectants (dimethyl sulfoxide (DMSO) or methyl glycol (Methyl)) were used. Analyzes of post-thaw sperm were performed as described for fresh sperm and additionally the fertilization rate analysis was performed. Fresh sperm presented a sperm concentration of 29.2 × 109 spermatozoa/mL, membrane integrity of 82.4%, and morphologically normal cells of 53%. After glucose activation (150 mM) a motility rate of 87.5%, sperm quality score of 5.0, and a duration of motility of 285 s were observed. For post-thaw sperm, MIII + Methyl and Androstar + Methyl solutions resulted in the highest motility rates of 40-48%. No differences were observed for motility duration, membrane integrity, and sperm morphology. Samples cryopreserved in Methyl (12-20%) showed a higher fertilization rate than DMSO, independently of the extender. In conclusion, the fresh sperm collected artificially from Chirostoma estor presents a compatible quality to carry out fertilization and can be cryopreserved in the commercial extenders MIII™ and Androstar Plus™ together with the cryoprotectant Methyl glycol.

Sex identification of the ornamental amazon fish Astronotus ocellatus by videoceloscopy and gonadal biopsy.

This study was conducted to evaluate and validate the efficacy and safety of videoceloscopy and gonadal biopsy as sexing methods for the A. ocellatus. A total of 31 adult individuals were used. Florfenicol (50 mg/kg) and morphine (5 mg/kg) were administered intramuscularly during the pre-surgical period. Animals were maintained in a supine position preceding a ventral midline incision and endoscope optics were then utilized for gonad visualization and sex identification. A gonadal fragment was collected using laparoscopic forceps and conditioned in 10 % formalin. To suture the cavity, polyamide yarn was used in a simple and continuous pattern. At 15 days subsequent to surgery, healing was evaluated, and the stitches were removed. Videoceloscopy accuracy and gonadal biopsy effectiveness were 97 % and 83 %, respectively. Total time devoted in the videoceloscopy, gonadal biopsy and surgery was longer for animals identified as males compared to females The survival rate was 100 %. There were differences regarding food consumption at 24 and 36 h post-surgery when compared to control specimens (pre-surgical) Regarding position in the water column, differences were observed at 24 and 72 h after surgery when compared individually to the control specimens. There were differences for interaction behavior at 24, 36 and 60 h, and regarding search for hiding places at 12 and 24 h after surgery in relation to the control specimens. The applied videoceloscopy and gonadal biopsy surgical techniques are, therefore, effective and safe for A. ocellatus sexing procedures.

Aceite de clavo como anestésico para el pez Pacu (piaractus mesopotamicus) / Clove oil as an anesthetic for pacu fish (piaractus mesopotamicus)

Con el propósito de minimizar el daño causado a los peces por el uso de sustancias anestésicas, se ha evaluado, en este trabajo, la concentración más adecuada de aceite de clavo como anestésico para el pez pacu (Piaractus mesopotamicus). Fueron utilizados 40 peces, con peso promedio de 716,18 g. Los tratamientos fueron compuestos de soluciones con diferentes concentraciones de aceite de clavo, 30, 60, 90, 120 y 150 mg/L. Fueron utilizados dos acuarios de vidrio: uno con 20 litros de agua con la solución anestésica para cada tratamiento y otro con 20 litros de agua limpia, para el regreso de los animales de la condición de sedación. Se observaron de forma individual: el tiempo hasta llegar a la anestesia profunda y el tiempo de recuperación de la anestesia profunda. El diseño experimental fue completamente aleatorizado con cinco tratamientos (concentraciones de aceite de clavo) y ocho repeticiones (peces). Los tiempos hasta anestesia profunda e hasta la recuperación total de los peces fueron evaluados por regresión, con 5% de significancia. Se observó un comportamiento linear de los tiempos hasta la anestesia profunda y hasta la recuperación total de los peces a concentraciones crecientes de aceite de clavo. La concentración de 30 mg/L es suficiente para la inducción anestésica de los animales, por otra parte, las concentraciones más elevadas, hasta 150 mg/L, causaron inducciones aún más rápidas, sin pérdida en las condiciones para la recuperación de los peces(AU)

In order to minimize damage to fish caused by the use of anesthetic substances, this study was to evaluate the concentration of clove oil as an anesthetic most appropriate for pacu (Piaractus mesopotamicus). A total of 40 fishes, with average weight of 716.18 g. The treatments were: anesthetic procedures with solutions of different concentrations of clove oil, 30, 60, 90, 120 and 150 mg.L-1. We used two glass aquariums: one with 20 liters of water containing the anesthetic solution for each treatment and the other with 20 liters of clean water, the animals return to the condition of sedation. Were observed individually: the time to reach deep anesthesia and recovery time of deep anesthesia. The experimental design was completely randomized design with five treatments (concentrations of clove oil) and eight replicates (individuals). The data were submitted to regression, with 5% significance level. There was a linear time of anesthesia and return of fish to increasing concentrations of clove oil, and 30 mg.L-1 is sufficient for the induction of the animals, but higher concentrations of up to 150 mg.L-1, still show inductions faster, without loss in time and conditions for the recovery of fish(AU)

Parámetros morfométricos en el rendimiento de los componentes corporales de Tilapia del Nilo (Oreochromis niloticus) / Morphometric Measurements in the Yield of Body Component in Nile of Tilapia (Oreochromis niloticus)

Con el propósito de estudiar los efectos de los parámetros morfométricos en el rendimiento corporal de la tilapia del Nilo (Oreochromis niloticus), se realizó este trabajo en la Universidad Federal de Lavras-MG, Brasil. Fueron mantenidos 1000 juveniles de linajes Supreme y Chitralada, con peso inicial entre 0,5 y 1,0 g, en separado, en dos tanques de cemento (40 m3). Fueron retiradas muestras y las medidas del alto, ancho y perímetros de las regiones del primer radio de las aletas dorsal, anal y de la circunferencia anterior del pedúnculo caudal, así como el largo de la cabeza y longitud total, fueron obtenidas de 78 peces de linaje Supreme y 93 de Chitralada. Los peces fueron pesados y disecados para obtención de los rendimientos de escamas y aletas, vísceras, cabeza, canal, piel, filete y residuo. Fueron hechos los análisis de correlación, el agrupamiento de variables en el dendograma y la regresión múltiple de los datos por el método backward para el estudio de la variación en los rendimientos corporales. Los rendimientos de los componentes corporales, en general, están poco relacionados con las medidas morfométricas. Los rendimientos de la canal y filete fueron calculados por las ecuaciones de regresión: RCAR = 43,2426 + 79,6399 LC2/CP (R2=0,096) y RFSP = 36,028 – 1,7415 AC1/LC2 (R2 = 0,014). Los análisis de regresión indicaron que el ancho a nivel del primer radio de la aleta anal, con relación a largura padrón (LC2/CP) y la altura a nivel del primer radio de la aleta dorsal, con relación al ancho del primer rayo de la nadadera anal (AC1/LC2) contribuyeron para la variación, respectivamente, en los rendimientos de canal y filete (AU)

The aim of this experiment was to study the influence of the morphometric measurements on characteristics of weight and on body yields of Nile tilapia (Oreochromis niloticus), an experiment was conducted in the pisciculture sector of the Federal University of Lavras, MG. Fingerlings of the Supreme and the Chitralada strains, with an initial weight of 0.5 a 1.0 g, were stocked and reared separately in two ponds (40 m3). During fish growth, samples were taken in 78 individuals of the Supreme strain and 93 of the Chitralada. Measurements of height, width and perimeters on the regions of the first ray of the dorsal and anal fins and on the anterior circumference of the caudal peduncle as well as the head and standard lengths were obtained. In addition, fish were weighted and dissected for obtaining the weights and yields of scales, fins, viscera, head, carcass, skin, fillet and residue. The correlation and grouping analysis of the data obtained revealed a low association degree between the morphometric measurements with the yelds of body components. There was a similarity between head and standard lengths and between height and perimeter of each region. The regression analysis was accomplished for body yelds as related with the morphometric relationship by backward method. The relation between the first-ray anal fin width and CP (LC2/CP) and between the first-ray dorsal f n and LC2 (AC1/LC2) were related to carcass (RCAR) and fillet (RFSP) yields, respectively. These yields were estimated as, RCAR = 43.2426 + 79.6399 LC2/CP (R2=0.096) and RFSP = 36.028 – 1.7415 AC1/LC2 (R2 = 0,014). The morphometric relationship was that better explained the yeld of body components had have the measurements LC2/CP and AC1/LC2, respectively, of the carcass and fillet yelds (AU)