Effects of Hypoxia Stress on Survival, Antioxidant and Anaerobic Metabolic Enzymes, and Related Gene Expression of Red Swamp Crayfish Procambarus clarkii.

The red swamp crayfish Procambarus clarkii is the most reared shrimp in China, but it is often affected by hypoxia stress in the process of seedling culture and adult crayfish culture. The oxygen consumption rate and asphyxiation point of juvenile crayfish (1.17 ± 0.03 g) and subadult crayfish (11.68 ± 0.11 g) at different temperatures (20, 22, 24, 26, and 28 °C) were studied. The survival, glycolysis, and expression of antioxidant genes were compared under 24 h acute hypoxia stress (1, 2, and 3 mg/L) and normal dissolved oxygen (7.5 mg/L). The results showed that the oxygen consumption rate and asphyxiation point of juvenile and subadult crayfish increased with increasing temperatures (20-28 °C). At the same temperature, the oxygen consumption rate and asphyxiation point of juvenile crayfish were significantly higher than those of subadult crayfish (p < 0.05). Within 24 h, the three hypoxia stress environments did not lead to the death of crayfish, indicating that P. clarkii has a strong ability to adapt to hypoxia. Hypoxia stress significantly affected the activities of antioxidant and anaerobic metabolic enzymes and gene expression in juvenile and subadult crayfish. The activities of the superoxide dismutase (SOD), catalase (CAT), and lactate dehydrogenase (LDH) and the content of lactic acid (LD) in the hepatopancreas of juvenile and subadult crayfish in the hypoxia stress groups increased significantly. The expression levels of SOD mRNA, CAT mRNA, Hsp70 mRNA, and crustin 4 mRNA in the hepatopancreas of juvenile and subadult crayfish in the hypoxia stress groups were significantly higher than those in the control group (p < 0.05), and the higher the degree of hypoxia stress, the higher the expression of each gene. The results showed that the antioxidant system of juvenile crayfish was more sensitive to hypoxia environments, and hypoxia stress resulted in increased stress levels in juvenile crayfish and subadult crayfish.

Comparative Analysis of the Symbiotic Microbiota in the Chinese Mitten Crab (Eriocheir sinensis): Microbial Structure, Co-Occurrence Patterns, and Predictive Functions.

Symbiotic microorganisms in the digestive and circulatory systems are found in various crustaceans, and their essential roles in crustacean health, nutrition, and disease have attracted considerable interest. Although the intestinal microbiota of the Chinese mitten crab (Eriocheir sinensis) has been extensively studied, information on the symbiotic microbiota at various sites of this aquatic economic species, particularly the hepatopancreas and hemolymph, is lacking. This study aimed to comprehensively characterize the hemolymph, hepatopancreas, and intestinal microbiota of Chinese mitten crab through the high-throughput sequencing of the 16S rRNA gene. Results showed no significant difference in microbial diversity between the hemolymph and hepatopancreas (Welch t-test; p > 0.05), but their microbial diversity was significantly higher than that in the intestine (p < 0.05). Distinct differences were found in the structure, composition, and predicted function of the symbiotic microbiota at these sites. At the phylum level, the hemolymph and hepatopancreas microbiota were dominated by Proteobacteria, Firmicutes, and Acidobacteriota, followed by Bacteroidota and Actinobacteriota, whereas the gut microbiota was mainly composed of Firmicutes, Proteobacteria, and Bacteroidota. At the genus level, Candidatus Hepatoplasma, Shewanella, and Aeromonas were dominant in the hepatopancreas; Candidatus Bacilloplasma, Roseimarinus, and Vibrio were dominant in the intestine; Enterobacter, norank_Vicinamibacterales, and Pseudomonas were relatively high-abundance genera in the hemolymph. The composition and abundance of symbiotic microbiota in the hemolymph and hepatopancreas were extremely similar (p > 0.05), and no significant difference in functional prediction was found (p > 0.05). Comparing the hemolymph in the intestine and hepatopancreas, the hemolymph had lower variation in bacterial composition among individuals, having a more uniform abundance of major bacterial taxa, a smaller coefficient of variation, and the highest proportion of shared genera. Network complexity varied greatly among the three sites. The hepatopancreas microbiota was the most complex, followed by the hemolymph microbiota, and the intestinal microbiota had the simplest network. This study revealed the taxonomic and functional characteristics of the hemolymph, hepatopancreas, and gut microbiota in Chinese mitten crab. The results expanded our understanding of the symbiotic microbiota in crustaceans, providing potential indicators for assessing the health status of Chinese mitten crab.

The feeding habits of small-bodied fishes mediate the strength of top-down effects on plankton and water quality in shallow subtropical lakes.

The proliferation of small-bodied fishes in lakes is often accompanied by deterioration of water quality and ecosystem function. However, the potential impacts of different types of small-bodied fish species (e.g., obligate zooplanktivores and omnivores) on subtropical lake ecosystems in particular have been overlooked mainly due to their small size, shorter life spans and lower economic value. Therefore, we conducted a mesocosm experiment to elucidate how plankton communities and water quality respond to different types of small-bodied fishes, including a common zooplanktivorous fish (thin sharpbelly Toxabramis swinhonis) and other small-bodied omnivorous fishes (bitterling Acheilognathus macropterus, crucian carp Carassius auratus and sharpbelly Hemiculter leucisculus). During the experiment, the mean weekly total nitrogen (TN), total phosphorus (TP), chemical oxygen demand (CODMn), turbidity, chlorophyll-a (Chl.α) and trophic level index (TLI) values were generally higher in treatments where fish were present compared to treatments where fish were absent, but responses varied. At the end of the experiment, phytoplankton abundance and biomass and the relative abundance and biomass of cyanophyta were higher while the abundance and biomass of large-bodied zooplankton were lower in the fish-present treatments. Moreover, the mean weekly TP, CODMn, Chl.α and TLI values were generally higher in treatments with the obligate zooplanktivore, thin sharpbelly, when compared to treatments with omnivorous fishes. Also, the ratio of zooplankton to phytoplankton biomass was the lowest, and the ratio of Chl.α to TP was the highest in treatments with thin sharpbelly. Collectively, these general findings indicate that an overabundance of small-bodied fishes can have adverse effects on water quality and plankton communities and that small-bodied zooplanktivorous fishes likely induce stronger top-down effects on plankton and water quality than omnivorous fishes. Our results emphasise that small-bodied fishes should be monitored and controlled if overabundant when managing or restoring shallow subtropical lakes. From the perspective of environmental protection, the combined stocking of different piscivorous fish species that feed in different habitat types could be a way forward to control small-bodied fishes with different feeding habits, but more research is needed to assess the feasibility of this approach.

Effect of dietary protein levels on the growth, enzyme activity, and immunological status of Culter mongolicus fingerlings.

A 65-day growth trial was conducted to investigate the dietary protein requirements for Culter mongolicus fingerlings. Isolipidic and isoenergetic diets were formulated with five dietary protein levels (32%, 37%, 42%, 47%, and 52%). Each diet was assigned to triplicate groups of 70 C. mongolicus fingerlings (0.99±0.08 g). The results indicated that weight gain and specific growth rate (SGR) increased with increasing dietary protein levels up to 47%. The activities of intestinal trypsin and lipase were the lowest in the 32% protein and 52% protein groups, while amylase activity reduced markedly in the 47% protein group. These results suggest that different dietary protein levels may cause different transformations of nutrients. The activities of superoxide dismutase (SOD) and lysozyme were not affected by varying dietary protein levels, except for those in the 32% protein group. In contrast, the content of malondialdehyde (MDA) increased with increasing dietary protein levels and reaching a maximum in the 52% protein group, suggesting that MDA accumulation depends on the protein concentration and the potential oxidative stress. Taken together, based on the broken-line analysis of SGR, we recommended the optimum dietary protein for C. mongolicus fingerlings to be 48.97%~49.31%.

Manipulation of fish community structure effectively restores submerged aquatic vegetation in a shallow subtropical lake.

Fish community manipulation and regulation has been largely overlooked as a mitigation strategy for restoring submerged aquatic vegetation (SAV) in shallow lakes of the middle and lower Yangtze River Basin (MLYRB). An in-situ fish exclusion experiment and a large-scale lake manipulation were conducted to test the hypothesis that the reasonable removal of benthivorous and herbivorous fish would facilitate the restoration and reconstruction of SAV in shallow lakes within the MLYRB. The in-situ exclusion experiment was conducted from April to October in 2017. Electrofishing was used to remove benthivorous and herbivorous fish from the exclosures. SAV were then artificially planted in the same pattern and density in both exclosures and adjacent open sites, and responses were measured for seven consecutive months. The mean percent coverage and biomass of SAV in the exclosures increased quickly and remained significantly higher than those in open sites over the duration of the experiment. Water quality also improved as turbidity, chlorophyll-a, total phosphorus and total nitrogen in the exclosures remained significantly lower than those in the open sites. After the in-situ experiment, a larger scale manipulation of fish in the entire submerged macrophyte zone (SMZ) was implemented from 2017 to 2020. After removing more than 2/3 of the benthivorous and herbivorous fish biomass by October 2020 in the SMZ, both the species richness and spatial coverage of SAV increased from 2 to 9 and from 1.7% to 32.2%, respectively. Our results provided clear evidence that fish are strong regulators of SAV productivity and that their reasonable removal facilitates ecological recovery. Therefore, we propose that fish community manipulation as implemented in this study be given more attention in addition to the reduction of external nutrient loading when designing projects to restore SAV in shallow lakes of the MLYRB.

Etiological characteristics of "tail blister disease" of Australian redclaw crayfish (Cherax quadricarinatus).

In November 2019, an acute disease outbreak in Australian redclaw crayfish (Cherax quadricarinatus) occurred in a farm in Hubei, China, with a cumulative mortality rate of over 80%. One of the characteristic symptoms of the disease was blisters on the tail. This symptom is also common in diseased Procambarus clarkii every year in this country, but the causative agent has not been determined. This study analyzed the etiological characteristics of this disease. Bacterial isolation and identification combined with high-throughput sequencing analysis were conducted to obtain the microbiota characteristics in the hemolymph, hepatopancreas, and intestines. Results showed that this outbreak was caused by infection from Aeromonas hydrophila and Aeromonas veronii. The underlying cause was stress imposed on crayfish during transferring from outdoor pond to indoor pond because of temperature drops. Aeromonas infection caused remarkable changes in the structure of the microbial composition in the hemolymph, hepatopancreas, and intestines of the crayfish. The abundance of Aeromonas in the hemolymph of the sick crayfish was as high as 99.33%. In particular, KEGG metabolic pathway analysis showed that some antibiotic synthesis, enterobactin biosynthesis, and myo-inositol degradation pathways were abundant in healthy crayfish hemolymphs, which may be the mechanism of maintaining crayfish health. Conversely, inhibition of these pathways led to the disorder of microbiota structure, finally leading to the occurrence of diseases. To the knowledge of the authors, this study was the first to use high-throughput amplicon sequencing targeting the 16S rRNA gene to find the causative bacteria in aquatic animals. This protocol can provide more comprehensive and reliable evidence for pathogen identification, even if the pathogenic bacteria are anaerobes or other hard-to-culture bacteria.

Reproductive pattern and population dynamics of commercial red swamp crayfish (Procambarus clarkii) from China: implications for sustainable aquaculture management.

BACKGROUND: The red swamp crayfish, Procambarus clarkii (Girard, 1852), is one of the most promising freshwater species for aquaculture in China. Understanding its reproductive pattern and population dynamics is crucial for sustainable management, but there is currently a lack of fundamental knowledge of commercial P. clarkii populations. Therefore, the purpose of this study was to investigate the reproductive pattern and population dynamics of commercial P. clarkii throughout the yearly cycle. METHODS: A total of 2,051 crayfish (1,012 females and 1,039 males) were collected from March 2016 to February 2017 in the area of Selection and Reproduction Center of Crayfish. The reproductive pattern was evaluated by the gonadosomatic index (GSI), hepatosomatic index (HSI), ovarian development and fecundity. Growth, mortality rates and exploitation rate were estimated by electronic length frequency analysis by R package "TropFishR" based on data of cephalothorax length (CTL). RESULTS: Our results demonstrated that spawning activities of P. clarkii took place from September to November, with a mean fecundity of 429 ± 9 eggs per female. There were two recruitments yearly, a major one from October to November and a minor one from March to May. With respect to population growth, five growth cohorts were identified for both females and males. Crayfish grew faster but attained smaller asymptotic maximum CTL as indicated by higher growth coefficient (K), growth parameter index (Ø') and lower asymptotic CTL (Linf ). The estimates of total mortality rate (Z), natural mortality rate (M) and fishing mortality rate (F) were 1.93, 1.02, 0.91 year-1 for females and 2.32, 0.93, 1.39 year-1 for males, which showed that the mortality of male crayfish was mainly caused by fishing. The estimates of exploitation rate (E) indicated that male crayfish were overexploited, with the values of 0.47 and 0.60 year-1 for females and males, respectively. DISCUSSION: P. clarkii spawned from September to November while two recruitments were observed yearly. We inferred that some eggs, prevented from hatching by low water temperature in winter, were more likely to hatch in the next spring. Moreover, the fishing mortality rate was relatively high for males, which might be related to the males-directed selection during the reproductive period. The higher values of exploitation rate in our study confirmed that males P. clarkii were overexploited and were under high fishing pressure. We thus suggest reducing fishing intensity on immature crayfish and avoid sex selection during the reproductive period to improve the overall sustainability of commercial P. clarkii populations.

Impacts of hatchery-reared mandarin fish Siniperca chuatsi stocking on wild fish community and water quality in a shallow Yangtze lake.

Mandarin fish Siniperca chuatsi, a valuable piscivorous fish, have been stocked into many lakes in China since the 1990s. This study did the first attempt to evaluate the ecological effects of hatchery-reared mandarin fish stocking in the Yangtze River basin lakes. Our study demonstrated a significant change in fish community composition after mandarin fish stocking, but no fish extinction was observed. No significant difference was observed in the total density of 13 forage fish before and after mandarin fish stocking, but the total biomass showed a significant decline after mandarin fish stocking. Significant differences in length-frequency distributions were observed for Carassius auratus, Pseudorasbora parva and Toxabramis swinhonis captured before and after stocking mandarin fish. No significant change in habitat distribution was detected before and after mandarin fish stocking. A marked decline in total nitrogen and a slight decline in total phosphorus were observed while a slight increasing trend for Secchi depth was found after stocking. Our findings suggested that mandarin fish stocking can increase predation pressure on forage fish and subsequently optimize the food web structure. Also, mandarin fish stocking has the potential to improve water quality and may be a feasible strategy to alleviate eutrophication of shallow Yangtze lakes.

Species-specific variations in reproductive traits of three yellow catfish species (Pelteobagrus spp.) in relation to habitats in the Three Gorges Reservoir, China.

The reproductive biology of three yellow catfish congeners was studied in the Three Gorges Reservoir of the Yangtze River, China. We compared reproductive traits among the lentic, transitional, and lotic zones. A total of 4502 individuals of the three species was collected, and the sex ratio, size at 50% maturity, spawning season, fecundity, and egg size were determined. Results showed that populations inhabiting the lotic zone spawned earlier than those inhabiting the lentic zone. For the three species, fecundities were significantly higher for populations in the lotic zone than for those in the lentic and transitional zones (P < 0.05). Pelteobagrus vachelli (Richardson) and P. fulvidraco (Richardson) displayed an obvious trade-off between egg size and fecundity, whereas P. nitidus (Sauvage et Dabry) produced the largest eggs in the lotic zone. Sex ratios were significantly different among zones (P < 0.05, for each species), but the bias patterns were different. Sizes at 50% maturity of female P. nitidus and P. vachelli were the largest in the lotic zone and the smallest in the transitional zone, but was similar among zones for P. fulvidraco. Overall results suggest that the three yellow catfish species developed different reproductive traits among the three habitats in the TGR, whereas the variations reflected further interspecific differences. Our study indicates the importance of riverine habitats for the conservation of species of fish, even for species such as these eurytopic catfish inhabiting the upper reach of the Yangtze River. This study further suggests that species-specific responses should be considered when evaluating the influences of new hydropower projects, even for such closely related species of fish.

Acute Toxicity of an Emerging Insecticide Pymetrozine to Procambarus clarkii Associated with Rice-Crayfish Culture (RCIS).

This study aims to evaluate the acute toxicity of pymetrozine to juvenile Procambarus clarkii. Two 96-h toxicity tests were conducted to assess the lethal concentration 50 (LC50) values, behaviors, and histopathology (at 50% of the 96 h LC50) after pymetrozine exposure. The results showed high toxicity of pymetrozine to juvenile P. clarkii in a dose and time dependent manner, with a decreasing LC50 from 1.034 mg/L at 24 h to 0.479 mg/L at 96 h. The maximum allowable concentration (MAC) of pymetrozine for P. clarkii was 0.106 mg/L. Behavioral abnormalities were observed in pymetrozine-treated crayfish, such as incunabular hyperexcitability, subsequent disequilibrium, lethargy, and increased defecation. Significant lesions were observed in all pymetrozine-treated tissues, including: (1) in gill, hemocytic infiltration and 33.27% of epithelial cells lesions; (2) in perigastric organs, 64.37%, 29.06%, and 13.99% of tubules with lumen atrophy, vacuolation, and cell lysis, respectively; (3) in heart, 2.5%, 8.55% and 7.74% of hemocytic infiltration, vacuolization, and hyperplasia, respectively; (4) in stomach, 80.82%, 17.77%, 6.98%, 5.24% of cuticula swelling, vacuolization, muscle fragmentation, hemocytic infiltration, respectively; (5) in midgut, 7.45%, 10.98%, 6.74%, and 13.6% of hyperplasia, tissue lysis and vacuolation, hemocytic infiltration, muscle fracture; and (6) in abdominal muscle, 14.09% of myofiber fracture and lysis. This research demonstrates that pymetrozine is highly toxic to juvenile P. clarkii, with significant effects on mortality, behavior and histopathology at concentrations of ≤1.1 mg/L, while the estimated practical concentration of pymetrozine in rice-crayfish culture water was around 20 times lower than the calculated MAC.

Mixture Toxicity of Bensulfuron-Methyl and Acetochlor to Red Swamp Crayfish (Procambarus clarkii): Behavioral, Morphological and Histological Effects.

The mixture of bensulfuron-methyl and acetochlor (MBA) has been widely applied as a rice herbicide in China, but the mixture toxicity of MBA to aquatic organisms is largely unknown. The current study aims to investigate the acute effects of MBA to juvenile red swamp crayfish, Procambarus clarkii. Firstly, a 96 h semi-static exposure was conducted to determine the Lethal Concentration 50 (LC50) values at 24, 48, 72 and 96 h, as well as to assess the behavioral and morphological effects. A second 96 h exposure was conducted at an MBA concentration of 50% of the 96 h LC50 (72.62 mg/L) to assess the histological changes in the gill, perigastric organ, muscle, heart, stomach, and midgut. The results showed that MBA exhibited low acute toxicity with the 24, 48, 72 and 96 h LC50 values of 191.25 (179.37-215.75), 166.81 (159.49-176.55), 154.30 (148.36-160.59) and 145.24 (138.94-151.27) mg/L, respectively. MBA-exposed crayfish showed body jerk, belly arch, equilibrium loss, body and appendage sway, and lethargy; and the dead crayfish showed dark gray or grayish-white body color and separated cephalothorax and abdomen. At 72.62 mg/L, MBA exposure caused significant histopathological alterations, mainly including the cuticular and epithelial degeneration of all the gills; atrophy of tubule lumina and cellular vacuolation of the perigastric organs (61.15 ± 9.90% of the tubules showed lesions); epithelial hyperplasia (48.40 ± 9.00%), myocardial fibers and epithelial cell lysis (17.30 ± 2.01%), and hemocytic infiltration of the hearts; cuticular swelling (15.82 ± 2.98%) and vacuolate connective tissue (11.30 ± 2.47%) of the stomachs; atrophied bladder cell and fragmented longitudinal muscles (95.23 ± 4.77%) of the midguts; and slight myofibers fragmentation and lysis (7.37 ± 0.53%) of the abdominal muscles. Our results indicate that MBA can cause behavioral, morphological and histopathological effects on juvenile P. clarkii at relatively high concentrations, but its acute toxicity is low compared with many other common herbicides.

Maintaining economic value of ecosystem services whilst reducing environmental cost: a way to achieve freshwater restoration in China.

Freshwater fisheries are central to food security in China and this remains one of the most important priorities for the growing human population. Thus, combining ecosystem restoration with economics is pivotal in setting successful conservation in China. Here, we have developed a practical management model that combines fishery improvement with conservation. For six years, a ban on fertilizer and a reduction of planktivorous fish stocking along with the introduction of both mandarin fish Siniperca chuatsi and Chinese mitten crab Eriocheir sinensis was apparent in Wuhu Lake, a highly eutrophic lake located in the middle reaches of the Yangtze River. Annual fish yield decreased slightly after the change in management, whereas fisheries income increased 2.6 times. Mandarin fish and Chinese mitten crab accounted for only 16% of total fisheries production but for 48% of total fisheries income. During this six year period, water clarity increased significantly from 61 cm to 111 cm. Total nitrogen, total phosphorus and chlorophyll decreased significantly from 1.14 to 0.84 mg/L, 0.077 to 0.045 mg/L, and 21.45 to 11.59 µg/L respectively, and macrophyte coverage increased by about 30%. Our results showed that the ecological status of shallow lakes could be rapidly reversed from eutrophic to oligotrophic using simple biomanipulation, whilst maintaining fisheries economic value. It also offers a better approach to shallow fisheries lake management in Asia where traditionally the stocking of Chinese carp and use of fertilizers is still popular.

Rapid life-history diversification of an introduced fish species across a localized thermal gradient.

Climatic variations are known to engender life-history diversification of species and populations at large spatial scales. However, the extent to which microgeographic variations in climate (e.g., those occurring within a single large ecosystem) can also drive life-history divergence is generally poorly documented. We exploited a spatial gradient in water temperatures at three sites across a large montane lake in southwest China (Lake Erhai) to examine the extent to which life histories of a short-lived fish species (icefish, Neosalanx taihuensis) diversified in response to thermal regime following introduction 25 y prior. In general, warmwater icefish variants grew faster, had larger adult body size and higher condition and fecundity, but matured at smaller sizes. Conversely, coldwater variants had smaller adult body size and lower condition, but matured at larger sizes and had larger eggs. These life-history differences strongly suggest that key ecological trade-offs exist for icefish populations exposed to different thermal regimes, and these trade-offs have driven relatively rapid diversification in the life histories of icefish within Lake Erhai. Results are surprisingly concordant with current knowledge on life-history evolution at macroecological scales, and suggest that improved conservation management might be possible by focusing on patterns operating at microgeographical, including, within-ecosystem scales.

Factors influencing the grass carp gut microbiome and its effect on metabolism.

Gut microbiota have attracted extensive attention recently because of their important role in host metabolism, immunity and health maintenance. The present study focused on factors affecting the gut microbiome of grass carp (Ctenopharyngodon idella) and further explored the potential effect of the gut microbiome on metabolism. Totally, 43.39 Gb of screened metagenomic sequences obtained from 24 gut samples were fully analysed. We detected 1228 phylotypes (116 Archaea and 1112 Bacteria), most of which belonged to the phyla Firmicutes, Proteobacteria and Fusobacteria. Totally, 41335 of the detected open reading frames (ORFs) were matched to Kyoto Encyclopedia of Genes and Genomes pathways, and carbohydrate and amino acid metabolism was the main matched pathway deduced from the annotated ORFs. Redundancy analysis based on the phylogenetic composition and gene composition of the gut microbiome indicated that gut fullness and feeding (i.e. ryegrass vs. commercial feed, and pond-cultured vs. wild) were significantly related to the gut microbiome. Moreover, many biosynthesis and metabolism pathways of carbohydrates, amino acids and lipids were significantly enhanced by the gut microbiome in ryegrass-fed grass carp. These findings suggest that the metabolic role played by the gut microbiome in grass carp can be affected by feeding. These findings contribute to the field of fish gut microbial ecology and also provide a basis for follow-up functional studies.

Effects of growth hormone (GH) transgene and nutrition on growth and bone development in common carp.

Limited information is available on effects of growth hormone transgene and nutrition on growth and development of aquatic animals. Here, we present a study to test these effects with growth-enhanced transgenic common carp under two nutritional conditions or feeding rations (i.e., 5% and 10% of fish body weight per day). Compared with the nontransgenic fish, the growth rates of the transgenic fish increased significantly in both feeding rations. The shape of the pharyngeal bone was similar among treatments, but the transgenic fish had relatively smaller and lighter pharyngeal bone compared with the nontransgenic fish. Calcium content of the pharyngeal bone of the transgenic fish was significantly lower than that of the nontransgenic fish. Feeding ration also affected growth rate but less of an effect on bone development. By manipulating intrinsic growth and controlling for both environment (e.g., feeding ration) and genetic background or genotype (e.g., transgenic or not), this study provides empirical evidence that the genotype has a stronger effect than the environment on pharyngeal bone development. The pharyngeal bone strength could be reduced by decreased calcium content and calcification in the transgenic carp.

Increased food intake in growth hormone-transgenic common carp (Cyprinus carpio L.) may be mediated by upregulating Agouti-related protein (AgRP).

In fish, food intake and feeding behavior are crucial for survival, competition, growth and reproduction. Growth hormone (GH)-transgenic common carp exhibit an enhanced growth rate, increased food intake and higher feed conversion rate. However, the underlying molecular mechanisms of feeding regulation in GH-transgenic (TG) fish are not clear. In this study, we observed feeding behavior of TG and non-transgenic (NT) common carp, and analyzed the mRNA expression levels of NPY, AgRP I, orexin, POMC, CCK, and CART I in the hypothalamus and telencephalon after behavioral observation. We detected similar gene expression levels in the hypothalamus of TG and NT common carp, which had been cultured in the field at the same age. Furthermore, we tested the effects of GH on hypothalamus fragments in vitro to confirm our findings. We demonstrated that TG common carp displayed increased food intake and reduced food consumption time, which were associated with a marked increase in hypothalamic AgRP I mRNA expression. Our results suggest that elevated GH levels may influence food intake and feeding behavior by upregulating the hypothalamic orexigenic factor AgRP I in GH-transgenic common carp.

[Relationships between spatial distribution of two dominant small-sized fishes and submerged macrophyte cover in Niushan Lake of China].

By using a set of pelagic gillnets with eight mesh sizes, an investigation was made on the spatial distribution of small fishes in submerged macrophyte habitats in a shallow macrophytic lake (Niushan Lake) in the middle reach of Yangtze River in summer, 2005. The fish composition, abundance, and size structure were examined along a biomass gradient of the most dominant submerged macrophyte Potamogeton maackianus. A total of 1124 individuals from 13 fish species were caught during the study period. According to the abundance and occurrence, sharpbelly Hemiculter leucisculus and redfin culter Cultrichthys erythropterus were identified as the two dominant small pelagic fishes in the lake. There existed dome-like relationships between the fish species richness and Shannon diversity index and the submerged macrophyte biomass within its observed range. For the two dominant small fishes, their abundance was significantly positively correlated with macrophyte biomass, and the average sizes of the individuals of H. leucisculus and C. erythropterus were larger in un-vegetated habitat but smaller in heavily vegetated habitats, indicating that the young individuals tended to live in dense submerged macrophyte covers. Other two habitat factors, i. e., water depth and distance to shore, had little effects on the spatial distribution of the two fish species. It was inferred that P. maackianus cover should be the important refuge habitat for the two dominant small-sized fishes in Niushan Lake, and it would be necessary to protect or restore the submerged macrophyte covers including P. maackianus.