Dietary thyme essential oil (Thymus vulgaris) changes serum stress markers, enzyme activity, and hematological parameters in gibel carp (Carassius auratus gibelio) exposed to silver nanoparticles

Gibel carp (Carassius auratus gibelio) juveniles with an average weight of 8.73± 2.1 g were fed a basal diet (control) and three experimental diets (T200, T400 and T800), containing 200, 400 and 800 mgkg^-1, respectively of thyme essential oil (Thymus vulgaris) for six weeks. After six weeks feeding trial, both control and thyme essential oil-added feed groups were exposed to sub-acute toxicity of silver nanoparticles (Ag-NPs) for a period of 96 h. At 96 h after exposure, the number of erythrocytes (RBC), leucocyte (WBC), hematocrit (Hct), hemoglobin (Hb), the differential leukocyte count in blood and alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity, HDL-cholesterol, LDL-cholesterol, cortisol and lactate in serum were measured. The results revealed feeding a thyme essential oil diet significantly conferred resistance to oxidative stress with sub-acute toxicity of Ag-NPs. Thyme essential oil enhanced the RBC and WBC count, hematocrit and hemoglobin content, at both low (T400) and high doses (T800). Biochemical analysis showed that serum ALT and AST activities, and LDL-cholesterol in the thyme essential oil-added feed groups were significantly reduced with increasing dietary thyme essential oil. Plasma HDL-cholesterol was significantly elevated by a high-dose of dietary thyme essential oil (T800). Furthermore, cortisol and lactate levels in serum significantly decreased throughout the thyme essential oil-added feed groups compared to the control. In summary, the use of thyme essential oil at 400 and 800 mgkg^-1, as dietary supplements, has potential to decrease oxidative stress of gibel carp providing resistance to non-fatal effects of pollutant by Ag-NPs

A brief exposure to low pH prior to refrigerated storage reduces the motility and viability of goldfish sperm (Carassius auratus, Linnaeus, 1758)

Optimal conditions of short-term storage of fish sperm are important for successful artificial reproduction protocols or incubation of sperm in a cryoprotecting medium for successful sperm cryopreservation. The influence of acid or alkaline incubating conditions on short term storage of goldfish sperm was investigated in a series of experiments. Goldfish sperm was diluted an immobilizing solution (1: 3) and incubated for 1 h in acidic, pH 6.5 (AC group) or alkaline, pH 8.5 (AL group) conditions. Subsequently, the sperm of both groups was further diluted (1: 3) in the immobilizing solution and the pH was adjusted to pH 7.51 (±0.02) and 7.56 (±0.07) in the AC and AL group respectively. The samples were kept in sealed bags and stored at 4°C. Viability (%) and motility (%) was estimated in samples obtained daily for the next 4 days. The results indicate that even a brief exposure of sperm to acid acidic conditions prior to storage accelerated the decline of sperm viability and motility during refrigeration

Effects of Density Stress on Growth Indices and Survival Rate of Goldfish ( )

Abstract: To evaluate the effect of stocking density on survival and growth rate of goldfish (Carassius auratus), after 2 weeks adaptation to the new condition, 126 of fish with initial body weight 3.50±0.27 g were distributed randomly among 12 aquaria (76 liter) to stocking densities 6, 9, 12 and 15 fish per aquarium, respectively. The experiments were continued for 5 months in triplicates. Fish were hand-fed two times daily up to 3% body weight/day. Biometric characteristics of fish including total length (cm), weight (g), growth rate (g), specific growth rate (g/day), feed conversion ratio (FCR) and condition factor were determined every two weeks. After the end of the trial period, the average weight of fish in each group was recorded 14.79±2.61, 15.98±0.32, 33.15±1.47 and 14.2±0.30 g, respectively. With the increasing density of goldfish, growth indices, including secondary weight, weight gain, specific growth rate and feed conversion ratio showed a significant difference (P<0.05), but condition factor did not show significant different (P<0.05). Survival rate were not significantly different among different densities (P>0.05). The results of this study indicate that density is significantly affected the growth, but had no significant effect on survival

Dietary amino acids impact sperm performance traits for a catadromous fish, Anguilla anguilla reared in captivity

Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modifications of the content of this paper are prohibited.[EN] Little is known about the role of dietary amino acids on male reproductive performance and gamete quality in fishes. Thus, the objective of this study was to investigate how "enhanced" feeds (EH-4, EH-5, EH-6), with modified amino acid composition, and the standard on-growing diet (DAN-EX) impact body composition, milt biochemistry, and sperm performance in male European eel, Anguilla anguilla. The fatty acid composition of EH4, EH-5, and EH-6 was similar but differed to that in DAN-EX, while amino acid composition varied between all four diets. Diet did not influence organ-somatic indices (e.g. HSI, GSI), while males fed EH-4 were heavier than other groups. Arginine, alanine, and lysine were the most abundant amino acids in milt (>11%), followed by glycine, aspartic acid, valine, glutamic acid, and leucine ( >5.66%). Diet impacted milt arginine, serine, proline, methionine, and histidine levels. Specifically, males fed DAN-EX, EH-4, and EH-5 had the highest percentages of arginine, while males fed EH-4 to EH-6 had higher percentages of serine. Proline was most abundant in males fed DAN-EX, EH-5, and EH-6. Both methionine and histidine were detected at low percentages ( 0.5 mL) for fertilization procedures. Spermatocrit (43.1 +/- 1.80%) did not differ between the diets (ranged from 37.57 to 47.21%). Dietary regime had an impact on sperm motility, such that eels fed EH-5 and EH-6 had the greatest percentage of motile cells. In addition, fish fed EH-5 and EH-6 (or DAN-EX) had the fastest swimming sperm. Spermatogenic maturity index of hormonally treated eels varied within groups but did not differ between dietary treatment groups after 9 weeks of injections (ranged from 0.54 to 0.80). The most interesting amino acids to scrutinize from PCA plots were proline, histidine, and valine as well as lysine and arginine. Here, eels with highly motile sperm had milt with high relative proportions of proline, histidine, and valine, but were particularly low in lysine and arginine. Together, our findings add evidence that certain amino acids regulate milt biochemistry, and that male ejaculate traits may be promoted by amino acid intake. Further studies to evaluate effects of supplemented amino acid diets on fertilization ability and inter-linked early developmental stages are required.This study was funded by the Innovation Fund Denmark under grant agreements no. 5184-00093B (EEL-HATCH) and 7076-00125B (ITSEEL). 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An overview of the reproductive cycle of cultured specimens of a potential candidate for Mediterranean aquaculture, Umbrina cirrosa

The shi drum (Umbrina cirrosa), belonging to the Sciaenidae family of fish (Nelson et al., 2016) has attracted attention as it is an excellent diversification species in Mediterranean aquaculture to combat the low prices of the most cultured species due to the appearance of new producing countries in the Mediterranean area. In this manuscript we describe for the first time the reproductive cycle of captive shi drum specimens that are able to spontaneously spawn in captivity. Our data show that spermatogenesis started in one year old shi drum spe- cimens, although the fish were not mature enough to complete the reproductive cycle. Thus, three year old specimens showed multiple spawns with around 65% of the eggs fertilized. In mature males, levels of 11-ke- totestosterone (11KT), testosterone (T) and progesterone (P) were detected in serum and gonad, while 17β- estradiol (E2) was only detected in gonad. Testicular levels of E2 and P suggest that these hormones might have a role in spawning and in the subsequent spermatogonial proliferation. In mature females, however, P and E2 were detected in serum and gonad, while T and 11KT were only detected in gonad. The pattern shown by P in the ovary indicates a link between this hormone and ovarian renewal and a delay in vitellogenesis, which would explain why the vitellogenic stage of shi drum is quite short. The level of T and 11KT in the ovary suggests a role in oocyte maturation and spawning. The differences observed between sexes in the prevalence of some infection and the important economic losses produced by some vertically transmitted pathogens; have increase the at- tention to the interactions between the immune and reproductive systems. In this study we observed differences between sexes in the protease and bactericide activity levels. Thus, this manuscript represents the most complete overview to date of the shi drum reproductive cycle.Versión del editor2,04

The Development of a New Experimental Facility for Pyrolysis Liquid Biofuel (Bio-oil) Combustion

Bio-oil is made from pyrolysis of waste biomass, which provides a carbon-neutral combustion energy alternative. This study focuses on developing burners for use with bio-oil. The previous burner using bio-oil/ethanol mixtures had a small combustion chamber with no refractory lining. It was not feasible to stabilize flames of pure bio-oil and the pollutant emissions were relatively high. In this study, the burner is made more comparable to actual bio-oil burners by implementing a refractory-lined combustion chamber and increasing the size of the chamber to investigate the relationship between burner design and combustion performance of bio-oil. After making some modifications in the new burner configuration and adjusting the operating parameters, stable flames of pure bio-oil are achieved with no major nozzle coking problems, and almost no unburned hydrocarbons and low CO emissions (below 10 ppm) in the exhaust gases. The effects of operating conditions on the pollutant emissions are also investigated.M.A.S

The Development of a New Experimental Facility for Pyrolysis Liquid Biofuel (Bio-oil) Combustion

Bio-oil is made from pyrolysis of waste biomass, which provides a carbon-neutral combustion energy alternative. This study focuses on developing burners for use with bio-oil. The previous burner using bio-oil/ethanol mixtures had a small combustion chamber with no refractory lining. It was not feasible to stabilize flames of pure bio-oil and the pollutant emissions were relatively high. In this study, the burner is made more comparable to actual bio-oil burners by implementing a refractory-lined combustion chamber and increasing the size of the chamber to investigate the relationship between burner design and combustion performance of bio-oil. After making some modifications in the new burner configuration and adjusting the operating parameters, stable flames of pure bio-oil are achieved with no major nozzle coking problems, and almost no unburned hydrocarbons and low CO emissions (below 10 ppm) in the exhaust gases. The effects of operating conditions on the pollutant emissions are also investigated.M.A.S

Ovarian fluid and its impacts on spermatozoa performance in fish: A review

Factors such as gamete quality can profoundly affect fertility, but the spawning micro-environment that surrounds the spermatozoa and eggs during gamete contact has largely been neglected. In fishes, understanding these gametic interactions is crucial because each female creates a unique spawning environment by simultaneously expelling her distinct ovarian fluid (OF) along with an egg batch. In turn, OF has been shown to influence spermatozoa performance traits by modifying spermatozoa behaviors and fertilization outcomes. Here, we shed light on these gametic interactions by overviewing literature on OF and how it impacts spermatozoa performance traits. Fish OF is clear or has slight coloration and can constitute ≤10–30% of egg mass. Viscosity of the OF is ∼2- to 3-fold higher than water and its pH ranges 6.2 to 8.8. Osmolality of the OF is lower in freshwater (190–322 mOsmol/kg) than marine species (289–514 mOsmol/kg). Na+ (98.3–213.7 mmol/L) and Cl− (89.8–172.7 mmol/L) are predominant ions in OF, while K+ (1.7–19.3 mmol/L), Mg2+ (0.4–8.1 mmol/L), and Ca2+(0.5–9.7 mmol/L) ions are detected at lower concentrations. Protein levels can be high in OF and exhibit intra- and inter-species variation (54–826 mg/100 mL). Fish OF also contains a series of organic components and substances that enhance and/or attract sperm towards the vicinity of an egg. OF can also differentially impact sperm based on genetic relatedness of mates, male phenotype (i.e. alternative reproductive tactics), or geographic origin. To conclude, when testing further reproductive paradigms, we suggest a shift from classic spermatozoa activation medium (water only) to more natural spawning media, which encompass OF-spermatozoa interactions

Outcomes of in vitro fertilization with frozen-thawed sperm: An analysis of post-thaw recovery of sperm, embryogenesis, offspring morphology, and skeletogenesis for a cyprinid fish

Gamete cryopreservation causes cellular damage and death. This study develops cryopreservation techniques for Levantine scraper, and deciphers how early offspring development is affected when eggs are sired with fresh and frozen-thawed sperm. Cryopreserved sperm did not affect embryogenesis at 2- and 4-cell stages, but impaired embryonic development at 8-cell stage. Embryonic viability decreased at organogenesis, where only 34-49% of embryos showed viability with frozen-thawed sperm. Hatching success and percentage of normal hatched embryos declined when fertilized with frozen-thawed sperm. Considering only frozen-thawed cells the DMSO-5%, METH-5%, and METH-10% treatments yielded highest hatch, while METH-5% and PG-5% yielded the most normal hatched embryos. Larval spinal torsion was higher for fresh than frozen-thawed sperm, where larvae with spinal torsion showed vertebral fusion and shape alterations during exogenous feeding. Both fresh and cryopreserved treatments showed abnormalities in caudal skeleton, while rates of defective yolk-sacs were higher for cryopreserved sperm, where larvae with defective yolks showed oversized yolk extension. Percentage of larvae with defective heads/eyes were also higher for cryopreserved sperm. Results show how frozen-thawed sperm impairs embryonic/larvae development and identifies frequency and position of abnormalities. Future studies should investigate how sperm DNA damage may have caused these alterations. This article is protected by copyright. All rights reserved

Growth, survivorship, and predator avoidance capability of larval shortnose sturgeon (Acipenser brevirostrum) in response to delayed feeding.

Larval shortnose sturgeon, reared at 17°C, were subjected to delayed feeding treatments of 0, 5, 10, 15, 18, and 23 days post-yolk absorption to examine effects of food deprivation on growth, survival, swimming activity, and escape capabilities. Starvation affected growth and survival but despite degree of starvation, larvae were able to resume growth and experience high survivorship following feeding. Specific growth rate based on larval dry weight for the period directly following first feeding was highest for the day 15 and 18 delayed feeding treatments. There were no differences in survival between the 0 and 5 day treatments, however survival was reduced to 71.2%, 45.4%, and 28.8% for 10, 15, and 18 day delayed feeding treatments, respectively. Shortnose sturgeon had a point-of-no-return (PNR; 55.7% initiated feeding) at ~19 days (or 42 days post-fertilization) following the full absorption of yolk. Mean percent swimming activity and swimming speeds showed an interaction between delayed feeding treatment and larval age, such that no differences were detected at 1 and 6 days post-yolk absorption, while these swimming behaviors generally increased or spiked as feeding was delayed for 10, 15, and 18 days post-yolk absorption. At 23 days post-yolk absorption, only swimming speed increased for larvae that were denied food for 18 days. While there was an interaction between delayed feeding treatments and age for proportion of larvae exhibiting an escape response, generally, larvae from all feeding treatments exhibited a positive escape response. There were also interactions between delayed feeding treatments and age post-yolk absorption for mean and maximum escape speeds, such that less aggressive escape responses were typically detected the longer larvae were denied food. Our research suggests that larval shortnose sturgeon increase physical activity during periods of starvation to find a food patch while remaining vigilant but maybe not as capable to defend against a predatory attack as fed individuals