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). Butts IAE, was also supported by the USDA National Institute of Food and Agriculture, Hatch project 1013854. Gallego V has a post-doc grant from the MICIU (Juan de la Cierva-Incorporacion; IJCI-201734200).Butts, IAE.; Hilmarsdóttir, GS.; Zadmajid, V.; Gallego Albiach, V.; Stottrup, JG.; Jacobsen, C.; Krüger-Johnsen, M.... (2020). Dietary amino acids impact sperm performance traits for a catadromous fish, Anguilla anguilla reared in captivity. Aquaculture. 518:1-12. https://doi.org/10.1016/j.aquaculture.2019.734602112518Abd-Elrazek, A. M., & Ahmed-Farid, O. A. H. (2017). Protective effect of L-carnitine and L-arginine against busulfan-induced oligospermia in adult rat. Andrologia, 50(1), e12806. doi:10.1111/and.12806Akiyama, T., Shiraishi, M., Yamamoto, T., & Unuma, T. (1996). Effect of Dietary Tryptophan on Maturation of Ayu Plecoglossus altivelis. Fisheries science, 62(5), 776-782. doi:10.2331/fishsci.62.776Alavi, S. M. H., Pšenička, M., Policar, T., Rodina, M., Hamáčková, J., Kozák, P., & Linhart, O. (2009). Sperm quality in male Barbus barbus L. fed different diets during the spawning season. Fish Physiology and Biochemistry, 35(4), 683-693. doi:10.1007/s10695-009-9325-7Asturiano, J. F., Sorbera, L. A., Carrillo, M., Zanuy, S., Ramos, J., Navarro, J. C., & Bromage, N. (2001). Reproductive performance in male European sea bass (Dicentrarchus labrax, L.) fed two PUFA-enriched experimental diets: a comparison with males fed a wet diet. Aquaculture, 194(1-2), 173-190. doi:10.1016/s0044-8486(00)00515-9Asturiano, J. F., Perez, L., Garzon, D. L., Penaranda, D. S., Marco-Jimenez, F., Martinez-Llorens, S., … Jover, M. (2005). Effect of different methods for the induction of spermiation on semen quality in European eel. Aquaculture Research, 36(15), 1480-1487. doi:10.1111/j.1365-2109.2005.01366.xBaeza, R., Mazzeo, I., Vílchez, M. C., Gallego, V., Peñaranda, D. S., Pérez, L., & Asturiano, J. F. (2014). Effect of thermal regime on fatty acid dynamics in male European eels (Anguilla anguilla) during hormonally-induced spermatogenesis. Aquaculture, 430, 86-97. doi:10.1016/j.aquaculture.2014.03.045Baeza, R., Mazzeo, I., Vílchez, M. C., Gallego, V., Peñaranda, D. S., Pérez, L., & Asturiano, J. F. (2015). Relationship between sperm quality parameters and the fatty acid composition of the muscle, liver and testis of European eel. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 181, 79-86. doi:10.1016/j.cbpa.2014.11.022Baeza, R., Peñaranda, D. S., Vílchez, M. C., Tveiten, H., Pérez, L., & Asturiano, J. F. (2015). Exploring correlations between sex steroids and fatty acids and their potential roles in the induced maturation of the male European eel. Aquaculture, 435, 328-335. doi:10.1016/j.aquaculture.2014.10.016Bahadorani, M., Tavalaee, M., Abedpoor, N., Ghaedi, K., Nazem, M. N., & Nasr-Esfahani, M. H. (2018). 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(2013). Evaluation of Methods to Determine Sperm Density for the European eel,Anguilla anguilla. Reproduction in Domestic Animals, 48(6), 936-944. doi:10.1111/rda.12189Støttrup, J. G., Jacobsen, C., Tomkiewicz, J., & Jarlbaek, H. (2012). Modification of essential fatty acid composition in broodstock of cultured European eelAnguilla anguillaL. Aquaculture Nutrition, 19(2), 172-185. doi:10.1111/j.1365-2095.2012.00967.xStøttrup, J. G., Tomkiewicz, J., Jacobsen, C., Butts, I. A. E., Holst, L. K., Krüger-Johnsen, M., … Kaushik, S. (2015). Development of a broodstock diet to improve developmental competence of embryos in European eel,Anguilla anguilla. Aquaculture Nutrition, 22(4), 725-737. doi:10.1111/anu.12299Dayal, J. S., Ahamad Ali, S., Thirunavukkarasu, A. R., Kailasam, M., & Subburaj, R. (2003). Nutrient an

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

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

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

Associations between Ovarian Fluid and Sperm Swimming Trajectories in Marine and Freshwater Teleosts: A Meta-Analysis

Marine and freshwater spawning environments present fish sperm with unique challenges, but for both, gametes often signal prior to contact via biochemical interactions through maternally-derived compounds (i.e. eggs and ovarian fluid; OF). For example, when OF is incorporated into the fertilization environment, sperm have been observed to exhibit changes in swimming trajectories (e.g. motility and velocity), yet it remains unclear whether the presence of this OF consistently improves sperm performance. The objectives of this study were to determine the overall effect of OF on sperm performance using meta-analysis. Published literature was searched for studies comparing sperm motility and/or velocity in the presence and absence of OF. For each study, the log response ratios (lnRR) was calculated, in which positive values indicate improved performance in the presence of OF. For motility, the overall effect size was non-significant (lnRR = 0.09, CL = −0.06, 0.24), whereas velocity was positively affected by OF (lnRR = 0.10; CL = 0.04, 0.17). When segregated by environment, for freshwater species there was a significant positive effect of the OF on velocity (lnRR = 0.18, CL = 0.07, 0.29), which translated to an increase in velocity of 20%. In contrast, no effect was detected for velocity in marine species (lnRR = −0.01, CL = −0.02, 0.01). Overall, there is evidence that OF improves sperm performance, although spawning environment and/or taxonomic factors are likely to moderate these sperm-OF interactions. Together, these results further our understanding of natural reproductive processes governing sperm performance, mating systems, and fertilization dynamics

Ovarian inseminated sperm impacts spawning success in zebrafish, Danio rerio (Hamilton, 1822) even in the absence of a male stimulus

Abstract Reproductive obstacles have led scientists to develop novel techniques/technologies for artificial reproduction. We aimed to investigate the possibility of propagating zebrafish females using sperm ovarian lavage with and without presence of male stimulus. This experiment consisted of several treatments: traditional spawning approaches with females and wild-type males (AB♀ × AB♂); no males present with non-manipulated females (AB♀); no males present with females inseminated with NaCl into ovarian lobes [AB♀(inj.NaCl)]; no males present with females inseminated with sperm from transgenic males into ovarian lobes [AB♀(inj.Tg♂)]; non-manipulated females kept separately from wild-type males (AB♀|AB♂); females kept separately from wild-type males and inseminated with NaCl into ovarian lobes [AB♀(inj.NaCl)|AB♂]; and females kept separately from wild-type males and inseminated with sperm from transgenic males into ovarian lobes [AB♀(inj.Tg♂)|AB♂]. There were no released eggs in both negative control treatments (AB♀ and AB♀|AB♂). Egg production increased (ranged from 0 to 28.5 eggs/female) when females were injected in the presence [AB♀ (inj.NaCl) |AB♂] or absence of male stimulus [AB♀ (inj.NaCl) and (AB♀(inj.Tg♂)]. A further increase in egg production [relative to AB♀, AB♀ (inj.NaCl), and AB♀|AB♂] was detected when females were inseminated with pooled sperm from transgenic males in the presence of male stimulus [AB♀(inj.Tg♂)|AB♂; ranged from 2.5 to 55 eggs/female] or when using traditional spawning approaches (AB♀ × AB♂; ranged from 25 to 131 eggs/female). Females inseminated with sperm produced embryos, although no differences were detected when females were inseminated with pooled sperm from transgenic males in presence (11.8 ± 16.3%) or absence (average = 12.6 ± 9.2%) of male stimulus. Traditional spawning approaches produced the most eggs (81.2 ± 42.3 per female) and highest fertilization rate (81.3 ± 10.4)

Impact of sperm density on hatch success for channel catfish (Ictalurus punctatus) ♀ × blue catfish (Ictalurus furcatus) ♂ hybrid production

Hybrid catfish, the progeny of channel catfish (Ictalurus punctatus) females × blue catfish (I. furcatus) males, are in high demand by the United States aquaculture industry because they outcompete each of the parent species for pond and raceway culture. Unfortunately, fry production can be a limiting factor due to a lack of natural hybridization between the two species and the necessity to sacrifice males for artificial fertilization. Males are costly to maintain in hatcheries and take longer to mature than channel catfish males. Therefore, it is essential to use the minimum quantity of sperm that allows for the highest fry production. In this study, channel catfish eggs from 9 females were collected from two different locations and fertilized with six different sperm to egg ratios ranging from 1.0 × 103:1 to 1.0 × 105:1. Embryos were then incubated under common environmental conditions until hatch. Overall average hatch success ± SE using sperm to egg ratios between 1 × 103 and 1.0 × 105 ranged from 18.3 ± 4.3 to 48.8 ± 8.3%. Overall, there was a significant effect of sperm to egg ratio (P 1 × 104:1 sperm per egg had no significant improvement on hatch success. Both study locations indicated significant impacts of sperm to egg ratio (P < .0001) on hatch success, which ranged from 18.2% to 57.3% at location 1 and from 18.6% to 32.3% at location 2. For location 1, ratios higher than 1 × 104:1 sperm per egg yielded no increase in hatch. This density threshold was lower for location 2, where no improvement at ratios higher than 5 × 103:1 were detected. Thus, lower sperm densities may be optimal under certain conditions. Maternal effects were responsible for up to 93.6% of the hatch variability, indicating the importance of individual females and egg quality for fry production. Based on these findings, we suggest that implementing the 1.0 × 104:1 sperm to egg ratio will support the standardization of in vitro fertilization methodologies for hybrid catfish production. Simultaneously, sperm economy and hatch capacity can be maximized for each blue catfish male