Effects of taurine, cysteine and melatonin as antioxidant supplements to the freezing medium of Prochilodus brevis sperm.

Cryopreservation consist of a set of methods to preserve cells and tissues by drastically reducing the temperature. Among some undesired effects, cryopreservation might generate reactive oxygen species that lead to an increase of oxidative stress, causing damage to cells. This study aimed to test taurine, cysteine, and melatonin on the freezing of Prochilodus brevis sperm and assess its effects on post-thawed sperm quality. Sperm was collected and seven pools were formed (n = 7). They were diluted (1:9) in standard medium (5% glucose, 10% dimethyl sulfoxide and 5% egg yolk) supplemented or not (control) with taurine (0.3, 1.0, 3.16 or 10.0 mM), cysteine (0.3, 1.0, 3.16 or 10.0 mM) or melatonin (0.6, 1.12, 2.0 or 3.56 mM). Post-thawed sperm was evaluated for kinetic (total motility, velocities, and percentage of rapid cells), morphology and membrane and DNA integrity. Differences were found when melatonin was used as an antioxidant. For the variables rapid sperm and sperm velocities, 3.56 mM melatonin presented higher results than the control (melatonin 0 mM). Melatonin 2 mM was similar to 3.56 mM on rapid sperm, average path velocity (VAP) and curvilinear velocity (VCL). No difference was found between concentration 0 mM (control) and taurine treatments. As for cysteine, 0.3 mM presented the best results for rapid sperm than 10 mM, and higher VCL and VAP than 1 mM. Melatonin 3.56 mM presented higher results on kinetic parameters (rapid motility, VCL, VSL and VAP) than other tested antioxidants. Therefore, melatonin 3.56 mM is recommended to be added to the sperm freezing medium of P. brevis.

Acute and subchronic effects of petroleum on the freshwater fish Hoplias aff. malabaricus / Efeitos agudo e subcrônico do petróleo no peixe de água doce Hoplias aff. malabaricus

Abstract Petroleum water soluble fraction (WSF) impairs organisms, but damages may vary among cell and tissue levels. The aim of the present study was to evaluate the acute (24 h, 48 h, 72 h) and subchronic effects (36 days) of WSF (0%, 25% and 100%) in juveniles of the Neotropical top predator fish Hoplias aff. malabaricus. The effects of WSF were evaluated at a molecular level using the comet assay and micronucleus test for genome damage; and at a morphological level through histological identification of liver pathologic lesions. In both acute and subchronic exposure we found low levels of DNA damage (< 10% of comet tail) and non-significant frequency of micronucleus in WSF exposed fish. The most significant liver lesions in WSF exposed fish were fatty vacuolization, hypertrophy and focal necrosis. Since these tissue injuries were progressive and persistent, their irreversibility may negatively affect fish recruitment, even in a such resistant top predator.

Resumo A fração solúvel de petróleo (WSF) prejudica os organismos, porém os danos podem variar entre os níveis celular e tecidual. O objetivo do presente estudo foi avaliar o efeito agudo (24 h, 48 h e 72 h) e subcrônico (36 dias) da WSF (0%, 25% e 100%) em juvenis do peixe neotropical predador topo Hoplias aff. malabaricus. Os efeitos da WSF foram avaliados no nível molecular utilizando o ensaio do cometa e o teste do micronúcleo para o dano genômico e no nível morfológico através da identificação histológica de lesões patológicas no fígado. Em ambas exposições (aguda e subcrônica) encontramos baixos níveis de dano no DNA (< 10% de DNA na cauda do cometa) e frequência de micronúcleos não significativa em peixes expostos a WSF. As lesões mais significativas no fígado dos peixes expostos a WSF foram a vacuolização lipídica, hipertrofia e focos de necroses. Como estas lesões foram progressivas e persistentes, sua irreversibilidade pode afetar negativamente o recrutamento dos peixes, mesmo sendo um predador topo resistente.

Supplementation of sulfate polysaccharides in the seminal cooling medium of common curimata (Prochilodus brevis).

BACKGROUND: The use of sulfated polysaccharides (PS) in seminal cooling is known to improve seminal quality. OBJECTIVE: To evaluate the effect of different concentrations of PS, extracted from the macroalgae Gracilaria domigensis as a supplement to the seminal cooling medium of the reophilic fish Prochilodus brevis (common curimatã). MATERIALS AND METHODS: Five semen pools were diluted in ACP-104 (treatment T1), in BTS® (T2) and in BTS® with different concentrations of PS (0.5 [T3]; 1.0 [T4] and 1.5 [T5]). The samples were cooled for different times (0, 6, 24, 48, 72, 96 and 120 h) and after each hour they were analyzed for: morphology, membrane integrity, DNA integrity and sperm kinetics. RESULTS: There were no significant differences between the treatments containing different concentrations of sulfated polysaccharides. Regarding the different cooling times, it was possible to observe that after hour 96, there was a reduction in the parameters of sperm kinetics. For DNA integrity there was no significant difference in relation to the treatments nor in relation to the hours. For membrane integrity, a reduction was noted as of hour 96, but there was no influence of polysaccharides. For the sperm morphology, there was no statistical difference between the hours, however the BTS was better than the ACP-104. CONCLUSION: It is concluded that the use of polysaccharides in seminal cooling has no negative effect on sperm parameters and proves that seminal cooling keeps the material viable for up to 72 hours. Doi: 10.54680/fr23410110512.

Conditions for transplantation of primordial germ cells in the yellowtail tetra, Astyanax altiparanae.

Biotechniques, including surrogate propagation derived from primordial germ cell (PGC) transplantation, are valuable tools for the reconstitution of endangered fish species. Although promising, there are no previous studies reporting such approaches using neotropical fish species. The aim of this study was to establish germline chimeras in neotropical fish by using the yellowtail tetra Astyanax altiparanae as a model species of the order Characiformes. Germline chimeras were obtained after transplantation of PGCs cultivated under different conditions: saline medium and supplemented with DMEM, amino acids, vitamins, glutamine, pyruvate, and fetal bovine serum, and subsequently transplanted into A. altiparanae triploids and triploid hybrids from the cross between A. altiparanae (♀) and A. fasciatus (♂). The results indicate ectopic migration in host embryos after transplantation of PGCs cultivated in saline medium. However, PGCs cultivated in supplemented medium migrated to the region of the gonadal ridge in 4.5% of triploid and 19.3% in triploid hybrid. In addition, the higher expression of dnd1, ddx4 and dazl genes was found in PGCs cultivated in supplemented culture medium. This indicates that the culture medium influences the maintenance and development of the cultivated cells. The expression levels of nanos and cxcr4b (related to the differentiation and migration of PGCs) were decreased in PGCs from the supplemented culture medium, supporting the results of ectopic migration. This is the first study to report the transplantation of PGCs to obtain germline chimera in neotropical species. The establishment of micromanipulation procedures in a model neotropical species will open new insights for the conservation of endangered species.

Distribution of metals in different environmental compartments and oxidative stress biomarkers in Bryconops caudomaculatus (Osteichthyes: Characiformes) from a bauxite mining area in the Eastern Amazon.

The Eastern Amazon is rich in bauxite ore. The extraction and processing of bauxite lead to the mobilization of Aluminum (Al) and other metals in environmental. We evaluated the metals (Al, Mn, Ba, and Cr) concentration in tissue, water, and sediment associated with antioxidant and oxidative damage responses in Bryconops caudomaculatus. The samplings were done in two hydrological periods (post-rain and post-dry periods) and at three points, located at two rivers: one in the surroundings of the mining area (P1) and other inside the mining area, upstream (P2), and downstream (P3). Defense antioxidant system biomarkers analyzed were total antioxidant capacity (ACAP) and glutathione-S-transferase (GST) activity. As an oxidative damage biomarker, the lipoperoxidation (LPO) was evaluated. Metals concentrations in the water and sediment were higher in the post-rain period compared to post-dry period. The water samples were acidic, with dissolved Al concentrations above the values established by local legislation at all points. In the gills, the metals accumulation was higher in fish from in the surrounding and upstream sites, and in the liver, was higher in fish from downstream site. Fish from the surrounding had increased antioxidant defenses, with higher ACAP in all tissues and higher GST in the gills. Consequently, they had lower levels of LPO. Fish from the mining area had decreased antioxidant defenses, with lower ACAP in all tissues and lower GST in the gills. Consequently, they had higher levels of LPO, indicating oxidative stress. The fish muscle was not responsive to GST and LPO at all sites. We conclude that the oxidative stress observed in the gills and liver of B. caudomaculatus from the area modified by the mining activity reflected the local anthropogenic impact status.

Seeds of Cucurbita maxima and Carica papaya are effective for controlling monogeneans in the gills of Leporinus macrocephalus.

This study was carried out to evaluate the anthelminthic efficacy of seeds of Cucurbita maxima and Carica papaya for controlling monogeneans in the gills of Leporinus macrocephalus, besides hepatosomatic and splenosomatic index and condition factor of host. The fish were fed with seeds of C. maxima or C. papaya for seven days, and these treatments did not cause any mortality among them. Jainus leporini, Urocleidoides paradoxus, Urocleidoides eremitus and Tereancistrum parvus were the monogeneans found, and their prevalence in fish fed with seeds of C. papaya was 100%, while in fish fed with C. maxima the prevalence was 42.8%. Fish fed with seeds of C. papaya showed decreased in intensity and abundance of monogeneans, while fish fed with seeds of C. maxima presented decreased in abundance. Feeding of L. macrocephalus with seeds of C. maxima or C. papaya had efficacy of 69.6 and 67.8%, respectively. The hepatosomatic index of fish fed with seeds of C. maxima or C. papaya was not affected by the treatments. However, the splenosomatic index and condition factor of fish fed with C. maxima seeds decreased. Seeds of C. maxima and C. papaya may be used for controlling monogeneans of L. macrocephalus in fish farming.

A prospective randomized controlled trial comparing the effect and safety of Piranha and VersaCut morcellation devices in transurethral holmium laser enucleation of the prostate.

PURPOSE: Holmium laser enucleation of the prostate (HoLEP) is reported to be widely used in the surgical treatment of benign prostatic hyperplasia (BPH), which consists of two procedures: enucleation and morcellation. This study is to examine the efficiency and safety of two different morcellator systems within a cohort of men undergoing HoLEP for BPH. METHODS: A total of 210 consecutive patients undergoing HoLEP and morcellation procedures were enrolled. Individuals were randomly divided into 2 separated groups: the first group (105 patients) was performed with a nephroscope using a mechanical Versacut morcellator and the second (105 patients) was performed with the new morcellation system, nephroscopes and Piranha morcellator. Perioperative characteristics were studied and analyzed. RESULTS: The morcellation time and the morcellation rate was similar when the prostate volume (PV) ≤ 60 mL while the morcellation time was significantly shorter and the morcellation rate was higher in the Piranha group with PV > 60 mL. No significant difference was observed according to the bladder irrigation time, indwelling catheter time, and discharge time. CONCLUSION: Piranha morcellator presents a higher efficiency for the prostate over 60 mL.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues.

The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

Acute and subchronic effects of petroleum on the freshwater fish Hoplias aff. malabaricus.

Petroleum water soluble fraction (WSF) impairs organisms, but damages may vary among cell and tissue levels. The aim of the present study was to evaluate the acute (24 h, 48 h, 72 h) and subchronic effects (36 days) of WSF (0%, 25% and 100%) in juveniles of the Neotropical top predator fish Hoplias aff. malabaricus. The effects of WSF were evaluated at a molecular level using the comet assay and micronucleus test for genome damage; and at a morphological level through histological identification of liver pathologic lesions. In both acute and subchronic exposure we found low levels of DNA damage (< 10% of comet tail) and non-significant frequency of micronucleus in WSF exposed fish. The most significant liver lesions in WSF exposed fish were fatty vacuolization, hypertrophy and focal necrosis. Since these tissue injuries were progressive and persistent, their irreversibility may negatively affect fish recruitment, even in a such resistant top predator.

Effects of hydroxy-selenomethionine on performance, innate immune system and antioxidant defense of tambaqui (Colossoma macropomum) exposed to a physical stressor.

Selenium (Se) is a mineral with natural antioxidant properties that constitutes a number of enzymes with a fundamental role in the immunity and antioxidant systems and may confer a protective role against oxidative stress in fish following exposure to physical stressors. Adopting an integrated approach, this study investigated simultaneously the role of hydroxy-selenomethionine (OH-SeMet) supplementation in performance, hematological parameters, innate immune, antioxidant capacity and tissue Se retention of tambaqui (Colossoma macropomum) and the possible protective role of dietary selenium when fish are exposed to a physical stressor (transport). Juvenile specimens (15.71 ± 1.90 g) were fed one of five diets: a basal unsupplemented diet (0.0 mg kg-1 Se) or diets supplemented with OH-SeMet to provide 0.3, 0.6, 0.9 and 1.2 mg kg-1 Se of diet for 75 days prior to subjection of fish to transport stress. Dietary supplementation with Se in the form of OH-SeMet for 75 days did not affect the production performance of juvenile tambaqui, but increased innate immunity parameters (oxidative burst) from the Se inclusion level of 0.6 mg kg-1 and induced the activation of the antioxidant defense system (GPX, GSH and GST) especially at the Se inclusion level of 0.9 mg kg-1. In addition, the Se content in the fillet rose significantly, as the OH-SeMet contents in the diet were increased. The stress caused by transport resulted in alterations in hematological parameters, blood protein profile and immune and enzymatic responses in the species. However, Se supplementation at 0.9 mg kg-1 had a positive effect, increasing innate immunity and activating antioxidant defenses (CAT and GPx, especially) after this physical stressor was applied. These results demonstrate that, when submitted to transport stress, juvenile tambaqui use Se stored in the muscle and dietary supplementation with OH-SeMet at 0.9 mg kg-1 improves the innate immunity and antioxidant system parameters of fish after transport. These findings reinforce the need for supplementing hydroxy-selenomethionine in commercial diets for tambaqui to ensure tissue Se reserves as a contingency in cases of stress.

Ovicidal effect of essential oils of Lippia alba, Lippia sidoides and Lippia gracilis on the acanthocephalan Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae).

Acanthocephalosis in tambaqui Colossoma macropomum culture is a limiting factor for the production of the species, being the most recurrent, which makes it necessary to develop strategies to control parasitosis even in the early stages of parasite development. This study aimed to elucidate the morphological characteristics of eggs of the acanthocephalan Neoechinorhynchus buttnerae and the ovicidal effect of the essential oils of Lippia alba, Lippia sidoides, and Lippia gracilis. The parasites were collected from hosts naturally infected with N. buttnerae, and the eggs obtained from the female 'parasites' abdominal cavity were added to 12-well plates containing the essential oils at different dilutions. After the eggs (N = 300) were separated and measured (length and width) with the aid of an optical microscope coupled to a digital program, the viability of the treated eggs was monitored 24 h after treatments, L. gracilis essential oil was 100% effective at the highest concentration, whereas L. sidoides essential oil showed over 50% efficacy at 2 mg/ml, reaching 100% efficacy at all higher concentrations. Lippia. alba essential oil did not show satisfactory efficacy in preventing N. buttnerae egg hatching.

Carbohydrate tolerance in Amazon tambaqui (Colossoma macropomum) revealed by NMR-metabolomics - Are glucose and fructose different sugars for fruit-eating fish?

In the present study, two approaches were followed to evaluate the metabolic responses of tambaqui (Colossoma macropomum), a frugivorous species, to intraperitoneal (IP) administration of glucose (GLU) and fructose (FRU) in fed (FED) and 10-day fasted (FAST) fish. Glucose and fructose tolerance tests were performed to assess the carbohydrate utilization and complementary NMR-metabolomics analyses were done to elucidate the impacts of sugar mobilization on the metabolic profile of plasma, liver and muscle. Blood was sampled from FED groups at 0, 3, 6 and 24 h; and at 0 and 24 h from FAST groups. Significant differences were observed in the hyperglycaemic peak between sugars at 3 h (GLU - 13.7 ± 2.0 mM vs. FRU - 8.7 ± 1.1 mM; saline 6.3 ± 0.6 mM) and on the return to normoglycaemia (GLU - 8.5 ± 2.2 mM vs. FRU - 5.2 ± 0.9 mM; saline 4.9 ± 0.6 mM) 6 h after IP on the FRU fish. The NMR-metabolomics approach allowed to conclude that tambaqui seems to be more responsive to the feeding regime (FED vs. FAST) than to the injected sugar (FRU vs. GLU). From the studied tissues, plasma showed no significant variations between feeding regimes at 24 h after IP, while muscle and liver revealed some variations on the final metabolome profile between FED and FAST groups. The metabolome variations between feeding regimes are indicative of changes on the amino acid utilization. Fish from FAST group seem to utilize amino acids as energy source rather than for protein synthesis and muscle growth. Variations on glucose concentration in muscle can also indicate different utilization of the sugars depending on the feeding regime.

Sulfated polysaccharides from seaweed as a supplement to Prochilodus brevis sperm freezing medium.

BACKGROUND: Using sulfated polysaccharides (SP) in fish sperm freezing medium promotes cell maintenance. OBJECTIVE: To evaluate the effect of different SP concentrations, extracted from two seaweeds (Gracilaria domingensis and Ulva fasciata), as a supplement to the sperm freezing medium of Prochilodus brevis. MATERIALS AND METHODS: Five semen pools were diluted in a solution composed of 5% glucose, 10 % dimethyl sulfoxide (DMSO) and different SP concentrations (0, 0.5, 1.0, 1.5, 2.0, 2.5 or 3.0 mg/mL). The samples were cryopreserved and, after 7 days, rewarmed and analyzed for morphology, plasma membrane integrity, DNA integrity, mitochondrial activity and sperm kinetics [total motility, progressive motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), and wobble (WOB)]. RESULTS: There was no interaction between seaweed and SP concentrations. Similar effects were observed with SP extracted from the two seaweeds, regardless of concentration. When comparing the SP concentrations, regardless of the seaweed, 1.0 mg/mL SP showed better results for VCL and VSL. For VAP and WOB, 1.0 mg/mL SP showed better results, but differed from 3.0 mg/mL. LIN followed the same pattern, but differed from SP at 2.5 and 3.0 mg/mL. For progressive motility, 1.0 mg/mL G. domingensis showed superior results compared to the control. For mitochondrial activity, G. domingensis was superior to U. fasciata, regardless of concentration. The lowest concentrations (0.5 and 1.0 mg/mL) showed the best results, regardless of the seaweed. However, the control was superior to all treatments tested. CONCLUSION: G. domingensis SP at the lowest concentrations might be a potential supplement to the P. brevis freezing medium. doi.org/10.54680/fr22210110412.

Dietary Euterpe oleracea Mart. attenuates seizures and damage to lipids in the brain of Colossoma macropomum.

The Amazonian açai fruit (Euterpe oleracea) has shown promising anticonvulsant properties, comparable to those of diazepam (BDZ) in in vivo models submitted to pentylenetetrazole (PTZ). PTZ is a classic convulsant agent used in studies for the purpose of screening anticonvulsants and investigating the mechanisms of epilepsy. Herein, we aimed to determine, for the first time, the effect of dietary administration of lyophilized E. oleracea (LEO) on PTZ-induced seizures, using juvenile Colossoma macropomum fish (9.1 ± 1.5 g) as a model. A control diet (0.00% LEO) and two levels of LEO inclusion were established: 5.00% and 10.0% LEO (w/w). Fish were divided into five groups (n = 5): control (0.9% physiological solution; i.p.), PTZ (PTZ 150 mg kg-1; i.p.), PTZ LEO 5.00%, PTZ LEO 10.0%, and BDZ-PTZ (BDZ: diazepam 10 mg kg-1; i.p.). In addition to the electroencephalography (EEG), the lipid peroxidation (TBARS) was quantified in the brain, along with the characterization of behavioral responses. Fish receiving PTZ showed intense action potential bursts (APB), which overlapped with a hyperactive behavior. In PTZ LEO 5.00% and 10.0% groups, convulsive behavior was significantly reduced compared to the PTZ group. Fish fed 5.00% or 10.0% LEO and exposed to PTZ showed less excitability and lower mean amplitude in tracings. The inclusion of 10.0% LEO in the diet prevented the increase in mean amplitude of the EEG waves by 80%, without significant differences to the quantified mean amplitude of the BDZ-PTZ group. TBARS concentration was reduced by 60% in the brain of fish fed 10.0% LEO-enriched diets relative to the PTZ-administered group. The results of this study demonstrated the anticonvulsant and protective roles of LEO to the brain, and the dietary inclusion of LEO seems to be promising for the formulation of functional diets. Results of this study may boost the interest on the anti-seizurogenic properties of Euterpe oleracea, including the development of new approaches for the prevention of seizures in humans and animals with low epileptic threshold.

Sperm characteristics, peroxidation lipid and antioxidant enzyme activity changes in milt of Brycon orbignyanus cryopreserved with melatonin in different freezing curves.

The objective of this study was to evaluate extender supplemented with melatonin and freezing curves on the antioxidant enzyme activity, peroxidation lipid and sperm characteristics of cryopreserved Brycon orbignyanus milt. Males (n = 16) and females (n = 5) were hormonally induced with two doses (0.5 mg and 5.0 mg kg-1) of carp pituitary extract, and their gametes were collected by light abdominal massage. The fresh milt was diluted at a ratio of 1:4 (milt:extender) in the following solutions: (Control) 10% methyl glycol (MG) + 5% Beltsville thawing solution; (M1) Control + 1 mM melatonin; and (M2) Control + 2 mM melatonin. The freezing curves were C1 (automated freezer) and C2 (dry shipper for 24 h). After each curve was recorded, the straws were transferred to a liquid nitrogen container until the analyses were performed. The samples were thawed in a water bath (60 °C, 8 s) and evaluated using the Sperm Class Analyzer software for the parameters total motility, progressive motility, curvilinear velocity, straight line velocity, mean displacement velocity, straightness, and linearity. The following were also measured: motility time, vitality, morphology, oxidative stress (lipid peroxidation, activity of catalase and superoxide dismutase, quantification of nitric oxide), and fertilization and hatching rates. The data were analyzed within R by one-way analysis of variance and Tukey's test for comparison of means (p < 0.05). A significant difference (p < 0.05) was observed between the solutions in vitality, morphology, motility, and fertilization rate, the solutions with melatonin having the best values. Total motility, progressive motility, and motility time were significantly different. Among oxidative stress markers, only lipid peroxidation and superoxide dismutase activity showed an effect of the curve × solution interaction (p < 0.05), the solutions with melatonin yielding the lowest values. The fertilization and hatching rates were also higher under the melatonin treatments, regardless of the curve. Melatonin 2 mM and slow curve are indicated for the cryopreservation of fish species sperm as it led to the slowest detrimental spermatozoa effects and better fertilization and hatching rates.

Ascorbic Acid Supplementation Improves Skeletal Muscle Growth in Pacu (Piaractus mesopotamicus) Juveniles: In Vivo and In Vitro Studies.

In fish, fasting leads to loss of muscle mass. This condition triggers oxidative stress, and therefore, antioxidants can be an alternative to muscle recovery. We investigated the effects of antioxidant ascorbic acid (AA) on the morphology, antioxidant enzyme activity, and gene expression in the skeletal muscle of pacu (Piaractus mesopotamicus) following fasting, using in vitro and in vivo strategies. Isolated muscle cells of the pacu were subjected to 72 h of nutrient restriction, followed by 24 h of incubation with nutrients or nutrients and AA (200 µM). Fish were fasted for 15 days, followed by 6 h and 15 and 30 days of refeeding with 100, 200, and 400 mg/kg of AA supplementation. AA addition increased cell diameter and the expression of anabolic and cell proliferation genes in vitro. In vivo, 400 mg/kg of AA increased anabolic and proliferative genes expression at 6 h of refeeding, the fiber diameter and the expression of genes related to cell proliferation at 15 days, and the expression of catabolic and oxidative metabolism genes at 30 days. Catalase activity remained low in the higher supplementation group. In conclusion, AA directly affected the isolated muscle cells, and the higher AA supplementation positively influenced muscle growth after fasting.

A sodium alginate bilayer coating incorporated with green propolis extract as a powerful tool to extend Colossoma macropomum fillet shelf-life.

Fish deterioration imposes great economic losses and serious human health hazards. The objective of this work was to evaluate the effect of a sodium alginate bilayer coating incorporated to the green propolis extract in shelf-life, physical-chemical properties, microbiological properties and sensory acceptance of Colossoma macropomum fillets. Additionally, the chemical composition, along with the antioxidant and antibacterial activities of Brazilian green propolis extract (GPE) were investigated. GPE showed promising antioxidant and antibacterial activities. Twenty-seven metabolites were identified by gas chromatography (GC-MS), which mainly comprised terpenoids (52.14%). Cyclolaudenol was the major constituent of the GPE and it is described for the first time in green propolis extracts. C. macropomum fillets treated with the sodium alginate bilayer coating showed high sensory acceptance, reduced microbial deterioration and extended shelf-life (up to 11 days) during cold storage. Taken together, these results show that GPE can be a great alternative of a natural preservative for fish coating.

Low temperature stress in a cultured fish (Piaractus mesopotamicus) fed with Pyropia columbina red seaweed-supplemented diet.

This study aimed to analyze the cold stress effects (in terms of hematology, energy reserves, and oxidative stress) in Piaractus mesopotamicus (pacú) and their mitigation by a Pyropia columbina red seaweed-supplemented diet. For this purpose, juvenile fish were fed with a control (CD) or a red seaweed-supplemented diet (RD) for 60 days, and then, the animals were exposed to a low temperature (14 °C) and a control temperature (24 °C) for 24 h. The cold shock generated an increase of hemoglobin levels in fish fed with both diets. In CD-fed fish, plasmatic triglycerides, cholesterol, and hepatic glycogen decreased after the thermal shock; meanwhile, the animals fed with RD showed decreased hepatic proteins, but increased cholesterol and hepatic glycogen. Regarding oxidative stress, antioxidant enzymes augmented their activity in the liver, intestine, and gills; meanwhile, lipid oxidative damage was observed in the liver and intestine of fish exposed to 14 °C and fed with both diets. Pacú was sensitive to cold shock, but no mitigation effects were observed in fish fed with the supplemented diet. Further research should target higher concentrations of P. columbina in supplemented diets to take advantage of this valuable resource.

Molecular characterization of different preproGnRHs in Astyanax altiparanae (Characiformes): Effects of GnRH on female reproduction.

Gonadotropin-releasing hormone (GnRH) is a key molecule in the initiation of the hypothalamic-pituitary-gonadal axis. Thus, knowledge about GnRH may contribute to the effectiveness of species reproduction. Using a Neotropical tetra Astyanax altiparanae as a fish model species, the GnRH forms were characterized at the molecular level and the role of injected GnRHs in vivo was evaluated. The full-length complementary DNA (cDNA) sequences of preproGnRH2 (612 bp) and preproGnRH3 (407 bp) of A. altiparanae were obtained, and the GnRH1 form was not detected. The cDNA sequences of preproGnRH2 and preproGnRH3 were found to be conserved, but a change in the amino acid at position 8 of the GnRH3 decapeptide of A. altiparanae was observed. All the injected GnRHs stimulated lhß messenger RNA (mRNA) expression but not fshß mRNA expression, and only GnRH2 was able to increase maturation-inducing steroid (MIS) levels and possibly stimulate oocyte release. Furthermore, only GnRH2 was able to start the entire reproductive hormonal cascade and induce spawning.

Effects of cold stress on juvenile Piaractus mesopotamicus and the mitigation by ß-carotene.

This study investigated the effects of cold stress on morphometrical and hematological biomarkers, energy metabolism, and oxidative stress in different tissues of P. mesopotamicus, and the protective role of ß-carotene. Fish were fed with a control diet (CD) and the same diet supplemented with 105 mg/kg ß-carotene (BD) for 60 days. After the feeding trial, fish fed CD or BD diets were exposed to control (24 °C) and low temperature (14 °C) for 24 h. Fish (CD and BD) exposed to thermal stress showed lower hepatosomatic index. The hemoglobin increased only in CD-fed fish exposed to 14 °C. Increased glycemia, plasmatic protein depletion, and decreased hepatic glycogen were observed in fish fed the CD, while only the lipid levels in liver were augmented in BD-fed fish exposed at 14 °C. Regarding the oxidative stress, increased antioxidant enzymes activity and lipid peroxidation were observed in CD-fed fish exposed to cold. The two-way ANOVA showed an interaction between dietary treatment and temperature for glucose and oxidative stress biomarkers, with the highest values recorded in 14 °C-exposed fish fed with the CD. Our study demonstrated that cold stress had the greatest impact on fish oxidative status, and ß-carotene reduces harmful effects induced by cold in P. mesopotamicus.