In vitro toxicology test system based on common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>) sperm analysis

The effect of heavy metals on the motility parameters of common carp sperm was investigated. In vitro test systems are widespread in ecotoxicology, and fish sperm can be a suitable model. For this reason, studies had been carried out in this topic; however, the published methods are not standard in several aspects (donor species, measured endpoint, etc.). In this study, a previously published toxicology-aimed sperm analysis protocol was tested to examine the effect of heavy metals (arsenic, cadmium, chromium, copper, mercury, nickel, zinc,) on common carp sperm. According to our results, PMOT is the most sensitive of the investigated parameters: dose-response was observed in case of each metal at low concentrations, already after 30 min of exposure. VCL was less sensitive: lower effects were observed at the same concentrations compared to PMOT. Among the examined parameters, LIN was the least affected: a dose-response was observed only in case of arsenic and mercury. The same sensitivity of motility parameters was observed on zebrafish sperm previously. Moreover, we found that PMOT, VCL, and LIN of common carp sperm were affected at the same concentrations as it had been observed in zebrafish, when the identical analytical protocol was applied. The only exception was As³⁺, where common carp sperm proved to be more sensitive: lower concentrations already reduced its motility parameters. Consequently, PMOT of common carp sperm is an accurate and fast bioindicator of aquatic pollution.     

Sperm cryopreservation of the noble Scallop Chlamys nobilis by a programmable freezing method: Effect of cryoprotectant

A study on Chlamys nobilis sperm cryopreservation by a programmable freezing method was conducted under laboratory condition. Four cryoprotectant agents (dimethyl sulfoxide [DMSO], methanol [MET], propanediol[PG] and ethylene glycol [EG]) and four concentrations (5%, 10%, 20% and 30%) were evaluated for their ability to retain sperm motility, movement characteristics and fertility. Results showed that cryopreserved sperm total motility produced by DMSO and MET at 5%, 10% and 20% were higher than other cryoprotectant treatment groups (CPA groups), as well as rapid sperm percentage. The curvilinear (VCL) and straight line (VSL) velocity produced by DMSO at 5% significantly higher than other CPA groups (p < 0.05), while no significant differences were found for average path (VAP) velocity. The lateral head displacement (ALH) in all CPA groups was similar and without significant difference (p > 0.05), as well as the beat‐cross frequency (BCF). A significant higher fertilization rate was produced in DMSO than that in MET at same concentration (p < 0.05), and no significant differences were found for differing concentrations of the same cryoprotectant (p > 0.05). Overall, 5%‐20% DMSO was more suitable for Chlamys nobilis sperm programmable cryopreservation when the calcium‐free Hanks’ balanced salt solution was used as the extender, and 10°C/min from 0°C to ?80°C was used as freezing rate. The findings presented in this study will benefit conservation programs for Chlamys nobilis.     

Fertilization strategies for Sea Bass Dicentrarchus labrax (Linnaeus, 1758): effects of pre‐incubation and duration of egg receptivity in seawater

Studying gamete biology can provide important information about a species fertilization strategy as well as their reproductive ecology. Currently, there is a lack of knowledge about how long sea bass Dicentrarchus labrax eggs can remain viable after being activated in seawater. The objectives of this study were to understand the effects of pre‐incubation of fresh and overripe sea bass eggs in seawater and to determine the duration of egg receptivity. Pooled eggs (fresh and overripe) from four females were pre‐incubated in seawater for 0 min (control), 0.5 min, 1 min, 3 min, 10 min and 30 min and then fertilized by pooled sperm from four males. The fresh eggs had a higher fertilization success than overripe eggs. Our results revealed a significant effect of pre‐incubation time for both the fresh (P < 0.01) and overripe eggs (P < 0.01). Fertilization success of eggs significantly declined for both these treatments after 3 min of pre‐incubation, which clearly indicates that sea bass eggs are able to be fertilized by sperm for up to 3 min after release into seawater. This study has particular importance for understanding fertilization strategies, reproductive potential, as well as reproductive ecology of sea bass.     

Assessment of different ionic adjustment strategies in low-salinity water on the growth of Litopenaeus vannamei and microbial community stoichiometry in a synbiotic nursery system

The aim of this study was to evaluate different ionic adjustment strategies in oligohaline water on the growth of Litopenaeus vannamei and C:N:P stoichiometric ratios of the floc microbial community (MC) in synbiotic nursery system. A 35-day culture (2000 PL's/m³) was carried out in 60 L units in a completely randomized experimental design, with three treatments: T1—seawater diluted to a salinity of 2.5 g/L (control), T2—water with a salinity of 2.5 g/L with potassium (K?) adjusted and T3—water at a salinity of 2.5 g/L with its Ca:Mg:K ratio adjusted to 1:3:1, each treatment in triplicates. The MC of flocs and the dissolved fraction (DF) was separated by filtration, where MC >1.6 µm and DF <1.6 µm. The juveniles reached a final weight of 0.40 ± 0.09–0.49 ± 0.04 g, survival above 80% and an average yield of 0.69 ± 0.18–0.81 ± 0.02 kg/m³, without significant differences among the treatments. A stabilizing trend of C:P, C:N and N:P ratios of MC was observed considering the variations of C:N:P in the DF, indicating a homeostatic behaviour of the floc MC, as occurs in systems with high nutrient availability. Our results indicate that the major ions initial concentrations (Ca²⁺: 25.07 mg/L, Mg²⁺: 89.75 mg/L and K?: 25.00 mg/L), total alkalinity 100.00 mg/L and total hardness 433.30 mg/L provide conditions that do not limit shrimp growth in oligohaline water synbiotic nursery system.     

Effect of vitamin E on spermatophore regeneration and quality of pond‐reared,black tiger shrimp (Penaeus monodon)

The objective of the present work was to evaluate the effects of different levels of vitamin E in feed on the spermatophore regeneration and quality of male Penaeus monodon. The experiment was carried out with the four following treatments: the basal diet no added vitamin E, the diet added 200, 600 and 1,000 mg/kg respectively. Spermatophore regeneration and quality were evaluated by spermatophore weight, sperm count and spermatophore absence rates, which male P. monodon were extruded spermatophore for feeding 20 and 40 days. In the experiment, the weight of the twice regenerated spermatophore of the males added to the vitamin E group was higher than that of the untreated control group. The weight of the first regenerated spermatophore with the addition of 1,000 mg/kg group was the highest and significantly higher than the control group (p < .05), but there was no significant difference among the three groups with different levels of vitamin E. The weight of the second regenerated spermatophore with the addition of 600 mg/kg group was the highest, followed by 1,000 mg/kg group, both of which were higher than the control group and the addition of 200 mg/kg group. Within the same group, the regeneration spermatophore weight showed overall upward trend as the feeding time, twice regenerate experiment spermatophore weight with added to the vitamin E groups were significantly higher than the initial value (p < .05), but three spermatophore weight of male shrimp at the control group had no significant difference. The sperm quantity and the percentage of normal sperm of the twice regenerated spermatophore of the males with added to the vitamin E group was higher than that of the untreated control group, and those of the addition of 200 mg/kg group was significantly higher than that of control group (p < .05). The total number of sperm and the percentage of living sperm of male shrimp in the experimental group decreased with the increase of vitamin E in the feed. Within the same group, the total number of sperm and the percentage of living sperm of male shrimp with added to the vitamin E groups showed overall upward trend as the feeding time and were significantly higher than the initial value (p < .05), but the control group was slightly down and had no significant difference. Comprehensive sperm weight, sperm quantity and living sperm percentage of three indicators, that adding 200 mg/kg of vitamin E in feed could effectively promote the spermatophore regeneration in the male P. monodon and improve the sperm quantity. The experimental results provide a scientific basis for the breeding of P. monodon.     

计算机辅助分析冷冻前后褐牙鲆精子运动特征

观察了褐牙鲆(Paralichthys olivaceus)精子在室温和低温下的活力与寿命,并应用计算机辅助精子分析系统(CASA)对超低温冷冻前后褐牙鲆精子的运动特征进行了分析,结果表明:褐牙鲆精子在室温(25℃)下,可存活4 d,在低温(4℃)下可存活7 d;鲜精的活力为(87. 74±5. 47)%,解冻后,精子的最高活性为(84. 00±3. 67)%;激活0. 5 min时,冻精与鲜精的运动精子占总精子数的百分率(MOT)无显著性差异(P>0. 05),但精子平均曲线运动速度(VCL)、平均直线运动速度(VSL)、平均路径运动速度(VAP)和精子运动路线的曲折程度(LIN)都有显著性差异(P <0. 05);激活4min和10min时,冻精与鲜精的MOT、VCL、VSL、VAP和LIN间都有显著性差异(P <0. 05)。鲜精激活0. 5 min后,直线运动、曲线运动、左右摆动和不运动的精子数目占总精子数的百分比分别为(24. 49±3. 87)%、(48. 53±4. 55)%、(24. 72±2. 86)%和(2. 27±1. 22)%;冻精激活0. 5min后,直线运动、曲线运动、左右摆动和不运动的精子数目占总精子数的百分比分别为(18. 58±1. 33)%、(35. 67±3. 00)%、(35. 24±2. 67)%和(10. 51±1. 33)%。随着激活时间的延长,褐牙鲆鲜精和冻精的运动状态均发生了改变,直线运动和曲线运动的精子数目逐渐减少,而不运动和左右摆动的精子数目逐渐增加。     

Minimally invasive evaluation of the anaesthetic efficacy of MS‐222 for ornamental discus fish using skin mucus biomarkers

Skin mucus has been demonstrated to provide stress biomarkers for evaluating the physiological status, providing new convenient and non‐invasive methods to detect stress response in fish. Here, we investigated the anaesthetic efficacy of tricaine methanesulphonate (MS‐222; 75–115 mg/L) for discus Symphysodon aequifasciata (34.27 ± 4.46 g; 8.10 ± 0.59 cm) using skin mucus stress biomarkers. The induction time, recovery time and respiratory frequency were also determined. According to the criteria for anaesthesia and recovery, discus fish to reach stage A3 (deep anaesthesia) within 3 min and to reach stage R4 (full recovery of normal behaviour) within 5 min were observed at 95–105 mg/L MS‐222. Respiratory frequency increased first and then decreased during MS‐222 exposure and increased after recovery. At 10 min after deep anaesthesia, a lower mucus glucose was only observed at 115 mg/L MS‐222. No change in mucus cortisol and increased lactate were observed in all treatments. Increased mucus protein was observed at 75, 85 and 95 mg/L MS‐222. At 10 min after recovery, increased mucus glucose and decreased mucus protein were observed at 85, 95 and 115 mg/L MS‐222, but increased mucus cortisol only at 115 mg/L and lactate only at 75 and 105 mg/L MS‐222. At 24 hr after recovery, mucus glucose returned to the initial level only at 75, 95 and 105 mg/L MS‐222, while cortisol at 75 and 85 mg/L and protein and lactate at 75 mg/L respectively. Overall, the effective dose of MS‐222 for discus fish has been suggested to be 95–105 mg/L.     

Biochemical responses of common carp,Cyprinus carpio,to transportation in plastic bags using thymol as a sedative agent

The present study aimed at assessing the effects of addition of thymol [0 (T0), 2.5 mg/L (T2.5), 5 mg/L (T5) and 10 mg/L (T10)] to water on biochemical and antioxidant responses of common carp, Cyprinus carpio, during 3-h transportation. The lowest dissolved oxygen and ammonia levels after transportation were related to T2.5 and T5 treatments respectively. T5 treatment exhibited lower cortisol/glucose responses after transportation and 24 h after that, whereas T2.5 and T10 exhibited elevations in these parameters at the same times. Hyperammonaemia and hyperchloraemia were observed in T2.5 treatment immediately after transportation, but such changes were observed in all treatments, 24 h after transportation. Addition of 10 mg/L thymol to water prevented uraemia after transportation, but all thymol-treated fish exhibited uraemia, 24 h after transportation. Thymol treatment failed to mitigate lactate elevation immediately after transportation; nevertheless, T2.5 and T10 had, respectively, the lowest and highest lactate levels, 24 h after transportation. T5 treatment inhibited alteration in superoxide dismutase activity, immediately after transportation, and T10 exhibited a similar effect, 24 h after transportation. Addition of 2.5 mg/L thymol to water significantly increased catalase activity immediately after transportation, but such a response was observed in all treatments, 24 h after transportation. T2.5 and T10 exhibited higher malondialdehyde levels than T0 immediately after transportation, but such a response was observed in all thymol-treated fish, 24 h after transportation. In conclusion, addition of 5 mg/L thymol is beneficial in reducing water ammonia concentration and stress in carp during transportation.     

Application of Computer‐assisted Sperm Analysis in Selecting the Suitable Solution for Common Carp,Cyprinus carpio L., Sperm Motility

The common carp, Cyprinus carpio L., sperm motility parameters were analyzed by using computer‐assisted sperm analysis system. The percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, µm/sec), average path velocity (VAP, µm/sec), the wobbling index (WOB, %), movement linearity (LIN, %), beat cross frequency (BCF, Hz), and amplitude of lateral head displacement (ALH, µm) were determined. Five activation solutions (As) were used to activate sperm movement. As 1 solution: 68 mM NaCl, 50 mM urea, 0.5% bovine serum albumin (BSA), pH: 7.7, 181 mOsm/kg; As 2 buffer: 100 mM NaCl, 10 mM Tris, 0.5% BSA, pH: 9.0, 199 mOsm/kg; As 3 solution: 86 mM NaCl, 0.5% BSA, pH: 7.4, 167 mOsm/kg; As 4 buffer: 5 mM KCl, 45 mM NaCl, 30 mM Tris, 0.5%, pH: 8.0, 160 mOsm/kg; and As 5 solution: distilled water with the addition of 0.5% BSA, pH: 7.3, <3 mOsm/kg. Among five tested solutions, a buffer with a pH of 9.0 and osmolality of approximately 200 mOsm/kg (As 2) was the most suitable. After its activation, a significant increase in MOT and ALH values was observed, which can be of importance to the effectiveness of egg fertilization .     

Effects of cysteine supplementation on the quality of cryopreserved sperm of South American silver catfish

The cryopreservation promotes cellular damage that could compromise sperm quality in terms of motility and fertility rates, which may be caused by oxidative stress. Thus, the aim of this study was to assess the effects of cysteine addition on post‐thaw sperm quality, DNA damage and indices of oxidative stress of the South American silver catfish (Rhamdia quelen) sperm, compared with the cryoprotectant solution without cysteine addition. Sperm collected from five males were cryopreserved in cryoprotectant solution (fructose 50 g/L, powdered milk 50 g/L and methanol 100 ml/L) containing different cysteine concentrations (0, 2.5, 5, 10 and 20 mM). After thawing, the following were measured: sperm motility, morphology, sperm viability, DNA damage, lipid peroxidation, concentration of carbonyl and sulfhydryl groups and the activity of SOD, CAT, GST and GPx enzymes. The lowest sperm motility was determined for semen cryopreserved with addition of 20 mM of cysteine. The control group had the lowest DNA damage and lipid peroxidation. The findings of this study show that cysteine addition had no positive effect on evaluated parameters. Therefore, the concentrations tested are not recommended for the supplementation of cryoprotectant solution for semen of R. quelen.     

Comparative study of the effects of heavy metals on embryonic development of zebrafish

This study aimed to investigate the effects of heavy metals, such as copper, zinc, iron, cobalt, chromium, aluminium, manganese and molybdenum, on zebrafish egg hatching rate and the subsequent survival rate of the larvae. Each metal was used in the various concentrations of0.05 mg/L, 50 mg/L and 500 mg/L to detect the threshold limit of tolerance. The study's results showed significant differences in the hatching and survival rate of larvae between the control group and most of the experimental groups when adding heavy metals in various concentrations. The highest concentration (500 mg/L) of the induced heavy metals tested in this study resulted in total egg or larval mortality within 96 hr and significantly lower egg or larval life expectancy. Among the tested metals, zinc and molybdenum had the most significant impact on hatching time, in contrast to iron that did not differentiate the results from the control group. Copper, cobalt, chromium, aluminium and manganese did affect zebrafish eggs depending on their exposure concentrations, which differentiated the results from the control group. Also, the study's findings demonstrate a significant dependence of the hatched egg heart rate on the metal concentration, and in particular, higher concentrations of metals correspond to higher heart rate. Zinc, Molybdenum and copper was proved to be the most toxic heavy metals causing increased hazard rate to mortality up to 96 hr and shorter life expectancy. Finally, skeletal deformities that occasionally observed did not significantly affect the total life expectancy of the hatched eggs.     

Fertilizing capacity and motility of tench Tinca tinca (L., 1758) sperm following cryopreservation

Experiments were carried out to develop an optimal cryopreservation protocol for tench sperm by testing the fertilizing capacity and motility parameters including progressive motility, curvilinear velocity (VCL) and linearity (LIN) of cryopreserved sperm. Three experiments were designed to this aim: first experiment where we tested the effects of two extenders (sugar‐based Grayling and ion‐based Kurokura 180) and two cryoprotectants (DMSO and methanol) on fertilization and hatching success; second where we tested the effect of cryoprotectant type (methanol or DMSO) in different concentrations (5%, 10% and 15%) on fertilization and hatching success; and third where we tested the effect of two cryoprotectants (methanol and DMSO) on sperm motility parameters (progressive motility, VCL and LIN) after 4 h post‐thaw storage (4°C). Sperm prepared with the sugar‐based Grayling extender displayed better fertilization and hatching rates independently of the applied cryoprotectant most likely due to glucose present which acted as an external cryoprotectant. Concerning cryoprotectant concentrations, the use of 10% methanol yielded the highest fertilization (85 ± 15%) and hatching (80 ± 13%) rates, which were significantly higher than in all other groups. During the post‐thaw storage time, 5% methanol, 10% methanol and 5% DMSO groups had significantly higher motility parameters than other groups and we observed no significant decline in any of the parameters during the storage time. Overall, we found that a sugar‐based extender in combination with methanol as cryoprotectant is suitable for the cryopreservation of tench sperm and allows storage of cryopreserved sperm for up to 4 h post thaw.     

Simultaneous effect of sex and dose on efficacy of clove oil,tricaine methanesulfonate, 2‐phenoxyethanol and propofol as anaesthetics in guppies,Poecilia reticulata (Peters)

We studied the simultaneous effect of sex and dose on anaesthesia efficacy to estimate, if possible, the lowest effective dose (LED) for clove oil, tricaine methanesulphonate (MS‐222), 2‐phenoxyethanol (2‐PE) and propofol in mature guppies. LED is the lowest dose needed to reach A5 stage in a mean time of 3 min, with mean recovery (R5) time of 5 min. We used four doses/anaesthetic: 25, 50, 75 and 100 mg/L for clove oil; 120,140,160 and 180 mg/L for MS‐222; 800, 1,000, 1,200 and 1,400 mg/L for 2‐PE, and 7.50, 8.25, 10.00 and 11.25 mg/L for propofol. Each dose was tested on 10 females and 10 males. Morbidity, mortality and behavioural changes were checked on two control groups (10 males and 10 females/group). Sedation (A3), A5 and R5 times were recorded. Significant interactive effect dose*sex on A5 time was found for each anaesthetic agent (p_dose*sex < .05). Except for MS‐222 (p_dose*sex = .284), significant interactive effect dose * sex on R5 time was found (p_dose*sex < .05). A5 time in females tended to be greater than in males, but, in general, R5 times were longer in males. Body size differences between males and females could explain these differences in MS‐222 on A5 time and for clove oil, 2‐PE and propofol on R5 time. No dose simultaneous meet LED′s conditions relating to both A5 and R5 times; therefore the lowest doses inducing A5 in a mean time of 3 min could be a safe guideline for anaesthetic procedure in both male and females.     

Trichlorfon is effective against Dawestrema cycloancistrium and does not alter the physiological parameters of arapaima (Arapaima gigas): A large Neotropical fish from the Amazon

This study investigated the in vitro and in vivo efficacy of trichlorfon against Dawestrema cycloancistrium, as well as its physiological effects on arapaima. Naturally parasitized arapaima gill arches were exposed in vitro to 100, 250, 500 and 750 mg/L of trichlorfon and a control group (only distilled water), in triplicate. Parasites were monitored, and mortality was used to determine the median effective concentration (EC₅₀). The 750 mg/L concentration demonstrated 100% in vitro efficacy against D. cycloancistrium after 60 min, while the intermediate (500 mg/L) and the lowest (100 and 250 mg/L) tested concentrations were completely efficient after 90 and 130 min, respectively. The EC_50-1h of trichlorfon for D. cycloancistrium was determined at 171.73 mg/L. Parasitized arapaima juveniles were exposed to a control group and 150 mg/L of trichlorfon in triplicate. Fish were exposed to two therapeutic baths for 60 min with 24-h intervals between treatments. Therapeutic baths with 150 mg/L of trichlorfon were 92.99% effective against D. cycloancistrium and did not bring about haematological alterations (erythrogram, white blood cell count, thrombogram, plasma glucose and total proteins). Therefore, 150 mg/L of trichlorfon can be used in therapeutic baths to control and treat D. cycloancistrium infestations with no physiological impairments for arapaima.     

Effect of Dimethyl sulfoxide (DMSO) and egg yolk on sperm motility,fertility and hatching rates of depik Rasbora tawarensis (Pisces: Cyprinidae) eggs after short‐term cryopreservation

Cryoprotectant is the crucial factor in the cryopreservation process. In general, there are two types of cryoprotectant, permeating and non‐permeating cryoprotectants. Dimethyl sulfoxide (DMSO) and egg yolk are common permeating and non‐permeating cryoprotectants respectively. Hence, the objective of the present study was to determine the best proportion of DMSO and egg yolk for the cryopreservation of Rasbora tawarensis sperm. A completely randomized experimental design was used in this study which involves two types of cryoprotectant and their combination at different concentrations, namely 5% DMSO, 5% egg yolk, 5% DMSO + 5% egg yolk and 2.5% DMSO + 2.5% egg yolk. Every treatment was conducted in three replicates. Combination of 5% DMSO + 5% egg yolk gave the best results cryoprotectant treatment had significant effects on sperm motility, fertilization and hatching rate of the R. tawarensis eggs (p < .05). It is concluded that the best proportion of cryoprotectants for sperm cryopreservation in this species is 5% DMSO + 5% egg yolk.     

Effects of elevated ambient histamine level on survival, growth, sexual maturity and tissue histamine accumulation of the mysis Neomysis awatschensis and Neomysis japonica Nakazawa

In Asia, trash fish have been routinely used in aquaculture often due to their local availability and lower costs compared to formulated feed. However, stale trash fish contain high levels of biogenic amines, which have been reported to be harmful to poultry and some aquatic animals. The present study elevated the effects of histamine levels in the water on survival, growth, sexual maturity and tissue histamine accumulation of two estuarine mysis, Neomysis awatschensis and Neomysis japonica Nakazawa. Newly hatched neonates of each mysis species were cultured in brackish water containing 0 (control), 5, 10 and 15 mg/L histamine until they reached sexual maturity. The results showed that compared to the control, survival of N. awatschensis was significantly lower in both 10 and 15 mg/L histamine treatments (69.5 and 62.5% vs. 86.5%). In the case of N. japonica Nakazawa, significantly lower survival was found for the 15 mg/L treatment (53.9% vs. 72.4%) (P < 0.05). In a separate experiment, neonates of each mysis species were cultured individually (15 neonates/treatment) to assess the sub-lethal effects of histamine. It was shown that at the level of 15 mg/L, histamine had a significant impact on body weight and length of sexual matured N. japonica Nakazawa, which were only 87.7 and 78.7% of that of the control. However, no significant difference in intermolt period was detected. In the case of N. awatschensis, no significant differences in both body weight and length among treatments were detected (P > 0.05) but the intermolt period was substantially prolonged at all histamine levels tested when compared to that of the control. Meanwhile, the timings required to reach sexual maturity for both male and female of the two mysis were generally increased with increasing histamine level, and such differences were often statistically significant (P < 0.05). Compared to the control, the tissue histamine concentration of the mysis increased significantly at histamine concentrations of 10 and 15 mg/L for N. japonica Nakazawa (P < 0.05). In the case of N. awatschensis, however, no significant differences were found among all treatments. Our findings indicate that histamine had clear negative effects on both mysis species although their responses and sensitivity appeared somewhat different. Our results suggest that further research is needed to assess the effects of water borne histamine on various aquatic animals.     

Evaluation of growth performance and lipid metabolism in zebrafish fed fructooligosaccharide using RNA sequencing

This study evaluated the effects of fructooligosaccharide (FOS) on growth performance and lipid metabolism in zebrafish (Danio rerio) by RNA sequencing. A total of 240 healthy zebrafish were randomly distributed into two groups. The control group was fed a basal diet, and the treatment group was fed a basal diet supplemented with 0.4% FOS. The results showed that there was no significant difference in growth performance (p > 0.05). The lipid content, total cholesterol, triglyceride, free fatty acid and low‐density lipoprotein were significantly lower (p < 0.05) in the liver of fish fed the 0.4% FOS diet than those of the fish fed the control diet, while the fish fed the 0.4% FOS diet had significantly higher (p < 0.05) high‐density lipoproteins than those of the control fish. Malic enzyme and fatty acid synthetase activities were significantly reduced by adding FOS. KEGG pathway analysis showed that the two pathways of steroid hormone biosynthesis and steroid biosynthesis were significantly enriched (p < 0.05). The profile of genes in these two pathways was affected by FOS in zebrafish. The results of the two pathways suggested new mechanisms underlying the lipid metabolism mechanism of FOS.     

Evaluation of sodium percarbonate as a bath treatment for amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a major health challenge for Atlantic salmon aquaculture globally. While freshwater bathing for 2 hr is effective in reducing infection severity, there is need for more rapid and lower cost alternatives. To this end, a combination of sodium percarbonate (SPC) in freshwater was examined for its treatment efficacy. Initial in vitro studies showed a reduction in amoeba viability when exposed for 30 min to freshwater containing >500 mg/L SPC. Subsequently, AGD‐affected salmon were bathed for 30 min in 16°C freshwater containing 100, 500 or 1,000 mg/L SPC, or for 2 hr in 16°C freshwater to mimic industry practice. Treatment at the highest SPC concentration caused extensive gill damage and substantial mortality. Neither occurred to a significant extent at lower SPC concentrations. Gill pathology of surviving fish 10 days post‐treatment (dpt) was comparable to or more severe than pre‐treatment, and significantly (p < .001) more severe than in 2 hr freshwater bathed fish. N. perurans DNA was confirmed by qPCR in all treatment groups at 10 dpt. The data indicate that a 30‐min exposure to SPC in freshwater is not a suitable alternative to existing freshwater treatment of AGD.     

The effects of the dietary biotin on zebrafish Danio rerio reproduction

An experiment was conducted to investigate the effects of dietary biotin on zebrafish Danio rerio reproduction. Biotin and avidin (biotin antagonist) were added to two isonitrogenous and isocaloric purified diets to provide molar avidin:biotin ratios of 0:1 (biotin‐sufficient diet) and 120:0 (biotin‐unsufficient diet). Each diet was fed to a group of males mean initial mass 0.262 g) and a group of females (mean initial mass 0.285 g) for 99 days. Following this feeding period, males fed biotin‐sufficient diet exhibited higher gonado‐somatic index, sperm density, sperm motility and sperm viability than those fed biotin‐unsufficient diet (P < 0.05). In the presence of biotin‐sufficient males, biotin‐sufficient females spawned more eggs (222.2 eggs) than biotin‐deficient females (18.8 eggs) (P < 0.05). The same pattern was observed with biotin‐deficient males (7.6 vs. 1.8 eggs) (P < 0.05). Biotin‐sufficient males generated a higher percentage of fertilized eggs (90% vs. 42%), hatching rate (62% vs. 27%), larvae survival (98% vs. 37%) and larvae length at 7 days post fertilization (4.4 mm vs. 4.2 mm) than biotin‐deficient males (P < 0.05) . Biotin status of the male is of high consideration for successful breeding in zebrafish, because it significantly impacts the reproductive performances of the female.     

Disinfection of almaco jack (Seriola rivoliana Valenciennes) eggs: Evaluation of three chemicals

Almaco jack (Seriola rivoliana Valenciennes) is an excellent candidate for aquaculture due to its fast growth rate and high market value. While S. rivoliana have adapted well to captivity, survival at early life stages can be improved to increase profitability during production. A wide range of variables cause larval mortalities but high bacterial loads in rearing tanks are often correlated with these losses. The aim of this study was to investigate the effect of egg disinfection on bacterial load and hatch rate of S. rivoliana. Disinfectants tested included formalin (F100 and F200; 100 and 200 mg/L, respectively, for 60 min), hydrogen peroxide (HPO; 300 mg/L for 10 min) and peracetic acid/hydrogen peroxide (PAA/HPO; 15.7 mg/L/39.6 mg/L for 1 min). Concentrations and contact times were determined based on current use in marine aquaculture and preliminary trials. Eggs treated with HPO and F100 had significantly higher hatch rates than the untreated control group. All treatments significantly decreased total Vibrio counts compared to untreated eggs; however, total bacterial counts were only decreased following treatments with PAA/HPO and F200. To prevent egg mortality due to bacterial overgrowth, consideration should be given to the use of surface disinfection using HPO or F100. Future studies should investigate the use of peracetic‐based products at lower doses.