Sea perch (Lateolabrax japonicus) UBC9 augments RGNNV infection by hindering RLRs-interferon response.

Small ubiquitin-like modifier (SUMO) is a reversible post-translational modification that regulates various biological processes in eukaryotes. Ubiquitin-conjugating enzyme 9 (UBC9) is the sole E2-conjugating enzyme responsible for SUMOylation and plays an important role in essential cellular functions. Here, we cloned the UBC9 gene from sea perch (Lateolabrax japonicus) (LjUBC9) and investigated its role in regulating the IFN response during red-spotted grouper nervous necrosis virus (RGNNV) infection. The LjUBC9 gene consisted of 477 base pairs and encoded a polypeptide of 158 amino acids with an active site cysteine residue and a UBCc domain. Phylogenetic analysis showed that LjUBC9 shared the closest evolutionary relationship with UBC9 from Paralichthys olivaceus. Tissue expression profile analysis demonstrated that LjUBC9 was significantly increased in multiple tissues of sea perch following RGNNV infection. Further experiments showed that overexpression of LjUBC9 significantly increased the mRNA and protein levels of RGNNV capsid protein in LJB cells infected with RGNNV, nevertheless knockdown of LjUBC9 had the opposite effect, suggesting that LjUBC9 exerted a pro-viral effect during RGNNV infection. More importantly, we found that the 93rd cysteine is crucial for its pro-viral effect. Additionally, dual luciferase assays revealed that LjUBC9 prominently attenuated the promoter activities of sea perch type Ⅰ interferon (IFN) in RGNNV-infected cells, and overexpression of LjUBC9 markedly suppressed the transcription of key genes associated with RLRs-IFN pathway. In summary, these findings elucidate that LjUBC9 impairs the RLRs-IFN response, resulting in enhanced RGNNV infection.

Environmentally relevant lead alters nuclear integrity in erythrocytes and generates oxidative stress in liver of Anabas testudineus: Involvement of Nrf2-Keap1 regulation and expression of biomarker genes.

Genotoxic and hepatotoxic effects of lead (Pb) on a freshwater fish, climbing perch (Anabas testudineus) were studied at an environmentally relevant concentration (43.3 ppm). The genotoxic potential of Pb was confirmed by micronucleus study, with increased frequencies of erythrocytic nuclear alterations like lobed, blebbed, notched, fragmented, and micronuclei were observed in erythrocytes in treated groups as compared to control. Inorganic Pb induces oxidative stress which is a consequence of elevated level of Reactive Oxygen Species. Hepatotoxicity was assessed both by the oxidative stress and cellular responses that emerged due to the toxic assault of Pb in the liver, the most important detoxifying organ. Upregulation of xenobiotic metabolizing enzyme like catalase was evident after 15, 30, and 90 days of exposure, and a profound effect was observed on 30th days. The level of lipid peroxidation and reduced glutathione was increased after Pb exposure. Histoarchitectural damages of liver were distinctly evident in treated fish. Western blot analysis confirmed the expressional alterations of stress-responsive marker proteins like Nrf2, Keap1, Hsp70, and Nqo1. Pb exposure resulted in increased expression of Hsp70, Nrf2, and Nqo1, whereas Keap1 was downregulated, suggesting the involvement of Nrf2-Keap1 regulation as a cytoprotective mechanism against Pb toxicity.

Brobdingnagians and Goliaths: two forms of gigantism in fish.

Two forms of gigantism are differentiated in fish, Brobdingnagian and Goliathan gigantism, the former applying to populations whose individuals are all larger than is typical for the taxon, the latter to single individuals within a population. While Brobdingnagian gigantism is largely explained by various ecological and evolutionary rules, Goliathan gigantism is not. A mechanistic hypothesis is proposed which explains Goliathan gigantism in terms of the reduction of oxygen requirements of individual fish via moving to cooler temperatures and/or acquiring larger, more energy-dense prey, which enable them to get bigger, and, in the process, sometimes generate bimodal size distributions that may qualify as gradual forms between Goliathan and Brobdingnagian gigantism. This mechanism, which relies on the manner in which their gill surface area grows, is more likely to operate in fish that can get big in the first place than in fish that remain small.

Feed form and perch design do not interact to production performance, gastrointestinal tract traits, behaviour and welfare of laying hens reared in enriched cages.

1. It was hypothesised that perch material and design may affect utility and maintenance energy demand in laying hens, affecting their feed form preferences and daily feed consumption. Accordingly, perch design and feed form on hen performance, gastrointestinal tract functions and some behavioural and welfare-related traits were studied in laying hens (ATAK-S) reared in enriched colony cages from 24 to 40 weeks of age.2. The experiment was a 2 × 2 factorial investigating two perch materials and design (circular steel or mushroom-shaped plastic) and feed form (mash or crumble). A total of 396 hens were randomly assigned to one of the four treatment groups with nine replicates each (11 birds per replicate).3. Except for feeding behaviour and prevalence of foot pad dermatitis at 40 weeks of age, the modification of the perch design did not have a significant effect on the traits examined. Mushroom-shaped plastic perches reduced feeding behaviour (p < 0.01) and the incidence of foot pad dermatitis at 40 weeks of age (p < 0.001).4. Performance traits were not affected by feed form. Intake, final body weight and FCR for crumble-fed laying hens were greater than those fed mash (p < 0.01).5. Hens fed mash had higher (p < 0.01) relative gizzard weights along with lower (p < 0.05) pH values, pancreatic chymotrypsin, amylase and lipase activities (p < 0.05), and duodenal absorption surface areas (p < 0.01). Ultimately, this gave higher protein digestibility (p < 0.05) compared to those receiving crumble.6. In conclusion, in enriched cage rearing systems, mashed feed was preferred over crumble to efficiently maintain productive performance. Compared to circular steel, plastic mushroom-shaped perches were associated with better footpad health and welfare.

Dietary supplementation of VA enhances growth, feed utilization, glucose and lipid metabolism, appetite, and antioxidant capacity of Chinese perch (Siniperca chuatsi).

The aim of this study was to investigate the effect of dietary vitamin A on juvenile Chinese perch (Siniperca chuatsi). Chinese perch were fed with five experimental diets containing 0, 20, 40, 60, and 80 mg VA·kg-1 for 8 weeks. Results showed that dietary vitamin A significantly influenced the fish's growth, feed utilization, glucose and lipid metabolism, appetite, and antioxidant capacity. Vitamin A-supplemented groups had higher weight gain rate (WGR) and specific growth rate (SGR) compared to the control group. Feed conversion ratio (FCR) was also lower in the vitamin A-supplemented groups. Dietary vitamin A had no significant effect on the survival rate (SR). Compared to the control group, fish fed with vitamin A had increased feed intake (FI), and the expression of appetite-promoting genes (npy and agrp) was significantly higher in the 40 mg VA·kg-1 group. Vitamin A also enhanced the utilization of dietary protein by Chinese perch. The serum glucose content of the fish fed with 40 mg VA·kg-1 diet was significantly higher than that of the control group and 20 mg VA·kg-1 diet, indicating that the promoting effect of VA on gluconeogenesis was greater than that on glycolysis. Additionally, dietary vitamin A increased the expression of lipid metabolism-related genes (hl and fas) and antioxidant genes (nrf2 and gpx) in the fish. These results suggest that the optimal vitamin A requirement of juvenile Chinese perch bream was estimated to be 37.32 mg VA·kg-1 based on broken-line regression analysis of WGR. In conclusion, this study provides valuable insights into the potential benefits of dietary vitamin A on the growth, metabolism, and antioxidant capacity of Chinese perch.

Dissolved oxygen and ammonia affect ammonia production via GDH/AMPK signaling pathway and alter flesh quality in Chinese perch (Siniperca chuatsi).

The dissolved oxygen (DO) and ammonia are crucial to the growth of Chinese perch (Siniperca chuatsi). Information on the effects of DO and total ammonia nitrogen (TAN) in regulating ammonia nitrogen excretion and flesh quality in Chinese perch is scanty. This study aimed to evaluate the effects of dissolved DO at oxygen levels of 3 mg/L and 9 mg/L, as well as the TAN concentrations of 0.3 mg/L and 0.9 mg/L on ammonia excretion and flesh quality. Results showed that the ammonia contents in plasma, muscle, and liver of the 9 mg/L DO group were significantly higher than those of the 3 mg/L DO group (P < 0.05). However, the expression of AMPK-related signaling pathway genes (gdh, lkb1, and ampd) and flesh quality indicators (gumminess, chewiness, hardness) in the 9 mg/L DO group were significantly lower than those in the 3 mg/L DO group. Under long-term exposure to 0.9 mg/L TAN, the ammonia contents in plasma and gill filaments, as well as muscle flesh quality (resilience, gumminess, chewiness, cohesiveness), were significantly lower than those in the 0.3 mg/L TAN group (P < 0.05). However, the activities of GDH and AMPD enzymes in the 0.9 mg/L TAN group were significantly higher than those in the 0.3 mg/L TAN group. In summary, when fish are exposed to 3 mg/L DO and 0.9 mg/L TAN in the environment for a long time, their amino acids are used for transamination and deamination, resulting in insufficient energy supply for Chinese perch, whereas 9 mg/L DO and 0.9 mg/L TAN caused deterioration of the flesh quality.

Addition of α-ketoglutaric acid (AKG) reduces deamination in Chinese perch (Siniperca chuatsi) fed with fermented soybean meal as a substitute for fishmeal.

To alleviate amino acid imbalances in fermented soybean meal as a replacement for fishmeal feeds, this study evaluated the effects of adding lysine (Lys), methionine (Met), and α-ketoglutaric acid (AKG) to fermented soybean meals for Chinese perch. Chinese perch (34 ± 3 g) were fed five diets for 66 days (fishmeal as the protein source of the basal diet [FM]; fermented soybean meal as a substitute for 30% fishmeal in the soybean meal diet [FSM]; addition of crystalline Lys and Met [AA]; addition of α-ketoglutaric acid [AKG]; and simultaneous addition of crystalline Lys, Met, and AKG [BA] to the soybean meal diet). At the end of the feeding trial, the FSM group had the highest feeding rate and the lowest weight gain rate among all the groups. The FM group had the highest protein retention and the lowest feed efficiency among the groups. The mRNA transcription level of genes related to the AMP-activating protein (AMPK) signaling pathway and amino acid response (AAR) signaling pathway (lkb1, atf4, and gcn2) were highest in the AA group (P < 0.05) but lower in the AKG and BA groups. In the AKG group, the mRNA transcription level of the gluconeogenesis pathway-related gene (pepck and g6pase) was significantly higher than that in the other four groups, but the mRNA transcription level of genes related to amino acid catabolism (gdh and ampd) was lower. Among all the groups, the FSM group had the lowest mRNA transcription level of genes associated with the mammalian target of rapamycin (mTOR) signaling pathway (mtor and s6k). These findings imply that the feeding rate of Chinese perch in the fermented soybean meal group was the highest, but the protein retention was the lowest, while the addition of Lys, Met, and AKG improved protein retention. In conclusion, the addition of AKG to fermented soybean meal as a fishmeal substitute reduced amino acid deamination, enhanced gluconeogenesis, and increased protein deposition, which contributed to the growth of Chinese perch, alleviated amino acid imbalances, and improved the feed utilization of Chinese perch.

Methyltransferase-like 3 suppresses red spotted grouper nervous necrosis virus and viral hemorrhagic septicemia virus infection by enhancing type I interferon responses in sea perch.

Methylation at the N6 position of adenosine (m6A) is the most abundant internal mRNA modification in eukaryotes, tightly associating with regulation of viral life circles and immune responses. Here, a methyltransferase-like 3 homolog gene from sea perch (Lateolabrax japonicus), designated LjMETTL3, was cloned and characterized, and its negative role in fish virus pathogenesis was uncovered. The cDNA of LjMETTL3 encoded a 601-amino acid protein with a MT-A70 domain, which shared the closest genetic relationship with Echeneis naucrates METTL3. Spatial expression analysis revealed that LjMETTL3 was more abundant in the immune tissues of sea perch post red spotted grouper nervous necrosis virus (RGNNV) or viral hemorrhagic septicemia virus (VHSV) infection. LjMETTL3 expression was significantly upregulated at 12 and 24 h post RGNNV and VHSV infection in vitro. In addition, ectopic expression of LjMETTL3 inhibited RGNNV and VHSV infection in LJB cells at 12 and 24 h post infection, whereas knockdown of LjMETTL3 led to opposite effects. Furthermore, we found that LjMETTL3 may participate in boosting the type I interferon responses by interacting with TANK-binding kinase. Taken together, these results disclosed the antiviral role of fish METTL3 against RGNNV and VHSV and provided evidence for understanding the potential mechanisms of fish METTL3 in antiviral innate immunity.

Effects of flow velocity on the growth performance, antioxidant activity, immunity and intestinal health of Chinese Perch (Siniperca chuatsi) in recirculating aquaculture systems.

The cultivation of Chinese Perch (Siniperca chuatsi) in recirculating aquaculture systems (RASs) has become a common trend. To explore the effect of flow velocity on the growth performance, antioxidant activity, immunity and intestinal health of Chinese Perch in RAS, 240 Chinese Perch with an initial weight of 70.66 ± 0.34 g were selected and randomly divided into 4 groups: control group [CK, 0 body length per second (bl/s)], low flow velocity (LF, 0.4 bl/s), middle flow velocity (MF, 0.8 bl/s) and high flow velocity (HF, 1.2 bl/s) for a 56-days experiment. The results showed that the flow velocity significantly increased the weight gain rate and feed intake in Chinese Perch. At 1.2 bl/s, the flow velocity increased the intestinal trypsin content and intestinal villus length. Furthermore, the relative expression of appetite-related genes showed a tendency to increase, and the relative expression of appetite-inhibiting genes had a significant decrease in HF. Regarding immune-related indicators, the activities of alanine aminotransferase (ALT) and aspartate transaminase (AST) were significantly higher in MF and HF. However, the activities of lysozyme (LZM) significantly decreased. Moreover, the activities of total superoxide dismutase (T-SOD) and catalase (CAT) were significantly higher in the CK group than in the other groups. Excessive flow velocity also caused the mRNA level of most immune-relevant genes to markedly decrease. With regard to intestinal health, the intestinal content sequencing results showed that MF could increase the intestinal diversity index of Chinese Perch. In addition, with increasing flow velocity, the relative abundance of Proteobacteria gradually increased, while the proportion of Firmicutes decreased. In conclusion, although the high flow velocity could promote growth, feeding, and digestion, inhibit fat deposition and increase the intestinal microbial abundance, the flow velocity caused stress, which leads to a decline in immunity and increases the death rate and the risk of intestinal disease in Chinese Perch. These findings provide theoretical support for the development of RASs for Chinese Perch.

Molecular characterization, transcriptional regulation of sea perch Moloney leukemia virus 10 and its antiviral function against VHSV.

Moloney leukemia virus 10 (MOV10) is a conserved RNA helicase and has multiple biological functions in mammals, but its role remains poorly understood in bony fish. Here, we cloned a MOV10 homolog from sea perch (Lateolabrax japonicus), which contained 23 exons and 22 introns, with an open reading frame of 3000 bp encoding 1000 amino acids. Tissue distribution analysis showed that MOV10 was high expressed in blood of sea perch. Promoter analysis revealed several putative multiple transcription factors binding sites, including upstream transcription factor 1, GATA-box, transcription initiation factor IIB, activator protein 1 and two interferon (IFN) stimulated response elements. Further analysis found that IFNc, IFNh, and IFNγ could not only activate IFN regulatory factor (IRF) 1 expression which in turn led to the induction of MOV10, but also prompted the expression of IRF10 to hinder excessive MOV10 expression. Moreover, IRF2 also suppressed MOV10 expression that was initiated by IRF1. Viral hemorrhagic septicemia virus (VHSV) infection upregulated MOV10 expression in vivo and in vitro, which in turn, enhanced IFNh expression and exhibited strong antiviral activity against VHSV proliferation. This study provides a basis to investigate the immune escape of VHSV by affecting the biological function of transcription factors in the signaling pathways associated with antiviral molecules.

Preparing for the future offspring: European perch (Perca fluviatilis) biosynthesis of physiologically required fatty acids for the gonads happens already in the autumn.

Long-chain polyunsaturated fatty acids (PUFA) are critical for reproduction and thermal adaptation. Year-round variability in the expression of fads2 (fatty acid desaturase 2) in the liver of European perch (Perca fluviatilis) in a boreal lake was tested in relation to individual variation in size, sex, and maturity, together with stable isotopes values as well as fatty acids (FA) content in different tissues and prey items. ARA and DHA primary production was restricted to the summer months, however, perch required larger amounts of these PUFA during winter, as their ARA and DHA muscle content was higher compared to summer. The expression of fads2 in perch liver increased during winter and was higher in mature females. Mature females stored DHA in their gonads already in late summer and autumn, long before the upcoming spring spawning period in May. Lower δ13CDHA values in the gonads in September suggest that these females actively synthesized DHA as part of this reproductive investment. Lower δ13CARA values in the liver of all individuals during winter suggest that perch were synthesizing essential FA to help cope with over-wintering conditions. Perch seem able to modulate its biosynthesis of physiologically required PUFA in situations of stress (fasting or cold temperatures) or in situations of high energetic demand (gonadal development). Biosynthesis of physiologically required PUFA may be an important part of survival and reproduction in aquatic food webs with long cold periods.

Swimming against the stream: A systems approach to rebuilding fishing stocks.

Solutions occurring within complex systems such as recovery of species are urgently needed. One path forward involves action agendas that extend across the full range of stakeholder groups. Approaches that can foster cooperative behavior across a range of vested interests can create environments supporting species recovery. This paper reports the stakeholder identification process used to gather divergent opinions. A total of 923 priorities from 239 stakeholders were identified. Consensus approaches were utilized, resulting in 25 top priorities, selected by 61 stakeholders. This study demonstrates how consensus areas for action can be illuminated and supported by diverse stakeholder groups whose relationships have previously been indicated as antagonistic. Results indicate support for a range of actions that can be implemented to protect iconic fish species such as pearl perch and snapper in Southeast Queensland. Stakeholders supported actions that; change fishing practices for groups (recreational, commercial and charter); extended monitoring and reporting; improved compliance; changes to fishing management practices; extended fishing control measures and the construction of additional artificial reef habitats.

Synthesis of spirulina loaded chitosan nanoparticles from prawn, Macrobrachium nipponense shell for extending the shelf life of pike-perch (Sander lucioperca) fillet during refrigerated storage.

BACKGROUND: This study was aimed to synthesize polymeric chitosan nanoparticles (CSNPs) from Macrobrachium nipponense shells using sodium triphosphate (TPP) as a crosslinker that was incorporated with spirulina extract (SPE) to improve the shelf life of pike-perch during refrigerated storage (4 °C). RESULTS: The encapsulation efficiency (EE) of SPE-loaded CSNPs decreased from 67% to 32%, and loading capacity (LC) was increased (10-14%) depending on their loaded SPE concentrations. The initial burst effect, followed by a slow-release at pH 7 (24 h), was observed. Free SPE and SPE incorporated CSNPs decreased microbial counts (total viable count, total psychotropic count, pseudomonas, and lactic acid bacteria) compared to control and unloaded CSNPs. Samples treated with free SPE or SPE-loaded CSNPs showed higher changes in odor, color, TVB-N (total volatile basic nitrogen), and TBA (thiobarbituric acid) compared with the unloaded CSNPs batch (P ≤ 0.05) until the tenth day of storage. However, fish fillets coated with SPE-loaded CSNPs had the highest overall consumer acceptability and the lowest values for TVB-N and TBA at the end of storage (14th  day). Controlled release of bioactive compounds in batches treated with SPE-CSNPs could delay the microbial degradation and enhance chemical reactions (TBA and TVB-N) in comparison to pure SPE during storage time. CONCLUSION: The incorporation of SPE in polymeric CSNPs can be considered as a promising material for controlled delivery of natural bioactive agents, and preservation of Pike perch quality during refrigerator storage. © 2022 Society of Chemical Industry.

Efficacy of hydrogen peroxide to reduce Gyrodactylus species infestation density on four fish species.

OBJECTIVE: The ability to effectively treat parasitic infestations of fish is of high importance for fish culture facilities. However, tools or approved therapies for treating infestations on fish are limited. This paper summarizes results from four separate clinical field studies that evaluated the efficacy of hydrogen peroxide (H2 O2 ; 35% PEROX-AID) for reducing Gyrodactylus spp. infestation density. METHODS: Three species of Gyrodactylus were studied (G. salmonis, hosts: Brook Trout Salvelinus fontinalis and Lake Trout S. namaycush; G. freemani, host: Yellow Perch Perca flavescens; G. hoffmani, host: Fathead Minnow Pimephales promelas) before and after the application of immersion H2 O2 therapy. RESULT: Parasite density was significantly reduced for each parasite × host combination to which H2 O2 therapy was applied. Two clinical field studies in salmonids were found to demonstrate substantial effectiveness that enabled 35% PEROX-AID approval. CONCLUSION: Further assessments of Gyrodactylus spp. could expand the use of H2 O2 for controlling these parasites in aquaculture. Specifically, H2 O2 was effective at all levels tested (50 or 75 mg H2 O2 /L for 60 min for the Yellow Perch and Fathead Minnow clinical field studies; 100 or 150 mg H2 O2 /L for 30 min regardless of salt pre-treatment for the Brook Trout study; and 100 mg H2 O2 /L for 30 min or 50 mg H2 O2 /L for 60 min for the Lake Trout study).

Sea perch (Lateolabrax japonicus) autophagy related gene 5 promotes RGNNV infection via inhibiting RLRs-interferon signaling pathway.

Autophagy-related gene 5 (Atg5), an essential component of autophagy machinery, is associated with innate immune responses. Here, the Atg5 of sea perch (Lateolabrax japonicus) (LjAtg5) was cloned and its role in regulating autophagy and interferon (IFN) response during red-spotted grouper nervous necrosis virus (RGNNV) infection was investigated. The LjAtg5 cDNA encoded a polypeptide of 275 amino acids with an APG5 domain, and had the closet genetic relationship with Micropterus salmoides Atg5. Autophagic detection showed LjAtg5 was conserved in inducing cell autophagy. Spatial expression analysis revealed LjAtg5 had a higher expression level in liver, brain, and kidney tissues of RGNNV-infected sea perch compared with the control group. In RGNNV-infected LJB cells, overexpression of LjAtg5 significantly increased the mRNA and protein levels of capsid protein, whereas knockdown of LjAtg5 led to the opposite effect, indicating LjAtg5 played a pro-viral role during RGNNV infection. Furthermore, dual luciferase reporter assay revealed LjAtg5 significantly suppressed the activation of sea perch type I IFN promoter in vitro, and overexpression of LjAtg5 strongly weaken the expression of genes related to the RIG-I-like receptors (RLRs) signaling pathway and IFN stimulated genes. These results suggested LjAtg5 promoted RGNNV infection by negatively regulating RLRs-IFN signaling pathway.

Altered Larval Yellow Perch Swimming Behavior Due to Methylmercury and PCB126 Detected Using Hidden Markov Chain Models.

Fish swimming behavior is a commonly measured response in aquatic ecotoxicology because behavior is considered a whole organism-level effect that integrates many sensory systems. Recent advancements in animal behavior models, such as hidden Markov chain models (HMM), suggest an improved analytical approach for toxicology. Using both new and traditional approaches, we examined the sublethal effects of PCB126 and methylmercury on yellow perch (YP) larvae (Perca flavescens) using three doses. Both approaches indicate larvae increase activity after exposure to either chemical. The middle methylmercury-dosed larvae showed multiple altered behavior patterns. First, larvae had a general increase in activity, typically performing more behavior states, more time swimming, and more swimming bouts per second. Second, when larvae were in a slow or medium swimming state, these larvae tended to switch between these states more often. Third, larvae swam slower during the swimming bouts. The upper PCB126-dosed larvae exhibited a higher proportion and a fast swimming state, but the total time spent swimming fast decreased. The middle PCB126-dosed larvae transitioned from fast to slow swimming states less often than the control larvae. These results indicate that developmental exposure to very low doses of these neurotoxicants alters YP larvae overall swimming behaviors, suggesting neurodevelopment alteration.

Cloning of gonadotropin Gph-alpha, FSH-beta and LH-beta subunits and seasonal profiles of steroid hormones in wild-caught Nile perch, Lates niloticus.

The Nile perch (np; Lates niloticus) is a freshwater teleost species with a potential for aquaculture in freshwater surroundings. However, wild-caught breeders have persistently failed to spawn spontaneously in captivity. Cloning of the gonadotropin subunits and analysing seasonal variation in reproductive hormone levels for a 1-year period were done to gain knowledge on the physiological basis underlying the reproductive biology of np. The ß-follicle-stimulating hormone (FSH-ß) and ß-luteinizing hormone (LH-ß) subunits and their common α-glycoprotein (Gph-α) subunit were cloned using 3' and 5' RACE-PCR. The nucleotide sequences of the npgph-α, npfsh-ß, and nplh-ß subunits were 664, 580 and 675 nucleotides in length, encoding peptides of 124, 120 and 148 amino acids, respectively. The deduced amino acid sequence of each mature subunit showed high similarity with its counterparts in other teleost. Sequence analysis showed that npFSH-ß is more similar to higher vertebrate FSH-ßs than to higher vertebrate LH-ßs. Heterologous immunoassay was calibrated to analyse pituitary LH levels. While the LH immunoassay showed parallelism of npLH with that of tilapia (ta), no parallelism for FSH was found. Levels of pituitary LH were higher in females at gonadal stages of vitellogenic oocytes, mature secondary oocytes and mature tertiary oocytes with migrating nucleus than in pre-vitellogenic oocytes and early and late perinucleolus oocytes. Using competitive steroid ELISA, variations in the levels of the steroid hormones 11-ketotestosterone (11-KT) in males and E2 in females were characterized in relation to month and reproductive index of Nile perch. Our findings show that in females, gonadosomatic index and plasma E2 were highly correlated (R2 = 0.699, n = 172) and peaked from September to November while in males, the gonadosomatic index and plasma 11-KT peaked from October to November. In female fish, both steroid hormones were detected in the plasma but greatly varied in concentrations. E2 in particular, increased with the developmental stage of the gonads. The levels of steroid hormones, E2 and 11-KT in females and males respectively increased with fish size (total lengths) and suggest that females mature at a body length of 40-59 cm than their counter part males that mature at a total length of 60-70 cm. Taken together, we describe seasonal endocrine differences in wild-caught adult Nile perch which could potentially be exploited to manipulate the reproductive axis in cultured breeders.

Dose-Dependent Cytotoxicity of Polypropylene Microplastics (PP-MPs) in Two Freshwater Fishes.

The massive accumulation of plastics over the decades in the aquatic environment has led to the dispersion of plastic components in aquatic ecosystems, invading the food webs. Plastics fragmented into microplastics can be bioaccumulated by fishes via different exposure routes, causing several adverse effects. In the present study, the dose-dependent cytotoxicity of 8−10 µm polypropylene microplastics (PP-MPs), at concentrations of 1 mg/g (low dose) and 10 mg/g dry food (high dose), was evaluated in the liver and gill tissues of two fish species, the zebrafish (Danio rerio) and the freshwater perch (Perca fluviatilis). According to our results, the inclusion of PP-MPs in the feed of D. rerio and P. fluviatilis hampered the cellular function of the gills and hepatic cells by lipid peroxidation, DNA damage, protein ubiquitination, apoptosis, autophagy, and changes in metabolite concentration, providing evidence that the toxicity of PP-MPs is dose dependent. With regard to the individual assays tested in the present study, the biggest impact was observed in DNA damage, which exhibited a maximum increase of 18.34-fold in the liver of D. rerio. The sensitivity of the two fish species studied differed, while no clear tissue specificity in both fish species was observed. The metabolome of both tissues was altered in both treatments, while tryptophan and nicotinic acid exhibited the greatest decrease among all metabolites in all treatments in comparison to the control. The battery of biomarkers used in the present study as well as metabolomic changes could be suggested as early-warning signals for the assessment of the aquatic environment quality against MPs. In addition, our results contribute to the elucidation of the mechanism induced by nanomaterials on tissues of aquatic organisms, since comprehending the magnitude of their impact on aquatic ecosystems is of great importance.

Season and species influence stable isotope ratios between lethally and non-lethally sampled tissues in freshwater fish.

The field of stable isotope ecology is moving away from lethal sampling (internal organs and muscle) towards non-lethal sampling (fins, scales and epidermal mucus). Lethally and non-lethally sampled tissues often differ in their stable isotope ratios due to differences in metabolic turnover rate and isotopic routing. If not accounted for when using non-lethal tissues, these differences may result in inaccurate estimates of resource use and trophic position derived from stable isotopes. To address this, the authors tested whether tissue type, season and their interaction influence the carbon and nitrogen stable isotope ratios of fishes and whether estimates of species trophic position and resource use are affected by tissue type, season and their interaction. This study developed linear conversion relationships between two fin types and dorsal muscle, accounting for seasonal variation. The authors focused on three common temperate freshwater fishes: northern pike Esox lucius, yellow perch Perca flavescens and lake whitefish Coregonus clupeaformis. They found that fins were enriched in 13 C and depleted in 15 N compared to muscle in all three species, but the effect of season and the interaction between tissue type and season were species and isotope dependent. The estimates of littoral resource use based on fin isotope ratios were between 13% and 36% greater than those based on muscle across species. Season affected this difference for some species, suggesting the potential importance of using season-specific conversions when working with non-lethal tissues. Fin and muscle stable isotopes produced similar estimates of trophic position for northern pike and yellow perch, but fin-based estimates were 0.2-0.4 trophic positions higher than muscle-based estimates for lake whitefish. The effect of season was negligible for estimates of trophic position in all species. Strong correlations existed between fin and muscle δ13 C and δ15 N values for all three species; thus, linear conversion relationships were developed. The results of this study support the use of non-lethal sampling in stable isotope studies of fishes. The authors suggest that researchers use tissue conversion relationships and account for seasonal variation in these relationships when differences between non-lethal tissues and muscle, and seasonal effects on those differences, are large relative to the scale of isotope values under investigation and/or the trophic discrimination factors under use.

Effects of barrier perch access and early dietary protein and energy dilution on some welfare parameters, tibiotarsus measurements, fear and mobility level in broiler chickens.

1. This experiment determined the effect of increasing mobility in broiler chickens by placing barrier perches between feeders and drinkers. In addition, the limitation of early weight gain by dietary energy and protein dilution on some welfare parameters, tibiotarsus measurements, fear and mobility level was examined.2. A total of 504 male, one-day-old broiler chickens (Ross 308) were randomly allocated to four treatments with three replicate pens per treatment and 42 broiler chickens per pen as a 2 × 2 factorial arrangement. Treatments included feeding the basal control diet between 0-42 days or a diet diluted by 10% energy and 20% crude protein fed between 0-21 d, with the control diet fed between 22-42 d. The second factor was the presence or absence of barrier perches. All treatments were allocated as a completely randomised design. Welfare parameters (foot pad dermatitis, hock burn, gait score, feather score, breast blister), tibiotarsus measurements (bone mineral content, bone mineral density, fluctuating asymmetry and relative fluctuating asymmetry), tonic immobility and mobility level were recorded.3. Results showed that access to a barrier perch and the diluted diet increased the mobility in broiler chickens. However, access to a barrier perch had no significant effect on tibiotarsus and welfare parameters. Broiler chickens had better gait scores (P < 0.05) and lower foot pad dermatitis incidence (P < 0.01) in groups fed the diluted diet. The diluted diet had no significant effect on bone mineral density but reduced the tibiotarsus bone mineral content (P < 0.05).4. In conclusion, the diluted diet provided positive effects in terms of leg health due to weight gain limitations in the early period, thus improving broiler chicken welfare.