ABSTRACT
In complex environments, fish often suffer from reduced physiological functioning due to starvation, which may have a significant effect on their behavioral adaptive strategies to predator attacks. We selected qingbo (Spinibarbus sinensis, which prefers flowing water habitats) and demasone cichlid (Chindongo demasoni, which prefers still water habitats), to investigate the differences in group distribution and dynamics between the two species when faced with a simulated predation attack under different trophic states (fasted for 2 weeks or fed). We chose to conduct our experiments in a six-arm maze that included a central area and six arms of equal length and width and to obtain evidence of how the fish used the various areas of the maze to respond to simulated predation attacks. We found that the two fish species differed in their responses to simulated predation attacks under different trophic states. The group structure of the two species was relatively stable, and the effect of fasting on the qingbo group was not significant, whereas the demasone cichlid group was more susceptible to the effects of fasting, shelter and a simulated predation attack. In an environment with shelter, both species had the same anti-predator strategy and tended to enter the shelter arm to hide after encountering a simulated predation attack. However, differences in the anti-predator strategies of the two species emerged in the no-shelter environment, with the qingbo tending to enter the arm to hide, whereas the demasone cichlid group chose to enter the central area to congregate, and this phenomenon was more pronounced in the fasted group. In conclusion, our research shows that even group-stable fish may shift their anti-predation strategies (i.e., entering a shelter to hide shifts to aggregating in situ into a shoal) when starved and that the worse the swimming ability of the fish, the more affected they are by starvation.
ABSTRACT
Understanding how animal collectives and societies form and function is a fundamental goal in animal biology. To date, research examining fish shoaling behavior has focused mostly on the general principles and ecological relevance of the phenomeon, while the ways in which physiological state (e.g., nutrition) affects collective behavior remain overlooked. Here, we investigated the shoaling behavior of common minnows (Phoxinus phoxinus) with three different nutritional states (control treatment: fasting for 24â¯h, fasting treatment: fasting for 7 days, and digestion treatment: 1â¯h after satiation feeding) across two ecological contexts (i.e., without and with food). No effects of either nutritional state or context were found on swimming speed, but the acceleration was greater in the digestion group than in the control group, with that in the fasting group being intermediate. Similar to change tendency in group length and group width of shoals, both interindividual distance and nearest neighbor distance were also greater in the fasting group than in the digestion group, suggesting that fasting and digestion may exert opposite driving forces on group cohesion. However, neither nutritional state nor context influenced the group area, group speed, group percent time moving, or group polarization. Both the foraging efficiency and the percentage of food items consumed by the fish shoals were greater in the fasting and control groups than in the digestion group. Our study suggested that one week of hunger and the energetically demanding stage of food digestion tend to have opposite influences on group shape, while the social foraging context does not influence the individual and group behavior of fish.
Subject(s)
Behavior, Animal , Nutritional Status , Social Behavior , Animals , Behavior, Animal/physiology , Nutritional Status/physiology , Cyprinidae/physiology , Feeding Behavior/physiology , Swimming/physiology , Fasting/physiologyABSTRACT
Heated effluent injection, cold hypolimnetic water inputs from dams, and extreme weather events can lead to unpredictable temperature fluctuations in natural waters, impacting fish performance and fitness. We hypothesized that fish exposed to such unpredictable fluctuations would exhibit weaker growth and enhanced thermal tolerance compared to predictable conditions. Qingbo (Spinibarbus sinensis) was selected as the experimental subject in this study. The qingbo were divided into a constant temperature group (C, 22 ± 0.5 °C), a predictable temperature fluctuation group (PF, 22 ± 4 °C, first warming, then cooling within a day) and an unpredictable temperature fluctuation group (UF, 22 ± 4 °C, the order of warming or cooling is random). After 40 days of temperature acclimation, the growth, metabolic rate, spontaneous activity, thermal tolerance, plasma cortisol concentration and liver hsp70 level of the fish were measured. Unexpectedly, neither the PF nor the UF group showed decreased growth compared to the C group. This could be attributed to the fact that temperature variation did not lead to a substantial increase in basic energy expenditure. Furthermore, feeding rates increased due to temperature fluctuations, although the difference was not significant. Both the PF and UF groups exhibited increased upper thermal tolerance, but only the UF group exhibited improved lower thermal tolerance and higher liver hsp70 levels compared to the C group. The qingbo that experienced unpredictable temperature fluctuations had the best thermal tolerance among the 3 groups, which might have occurred because they had the highest level of hsp70 expression. This may safeguard fish against the potential lethal consequences of extreme temperatures in the future. These findings suggested that qingbo exhibited excellent adaptability to both predictable and unpredictable temperature fluctuations, which may be associated with frequent temperature fluctuations in its natural habitat.
Subject(s)
Acclimatization , Temperature , Animals , Acclimatization/physiology , Thermotolerance , Hydrocortisone/blood , Hydrocortisone/metabolism , Liver/metabolism , Liver/physiology , HSP70 Heat-Shock Proteins/metabolism , Basal Metabolism , Energy MetabolismABSTRACT
Several studies have reported that animals' personalities are often correlated with individual differences in cognition. Here, we tested whether personality is related to cognition across species, focusing on 10 freshwater fishes and a task relevant for fitness, the ability to discriminate shoal size. Bolder species exhibited more 'shuttle' behavior for information sampling during shoal selection and showed high performance (HP) in the numerical discrimination than shyer species, i.e., low performance (LP) species. Species at both the high and low ends of sociability showed LP, possibly due to loosened selection pressure because of either no need to perform shoal size discrimination tasks frequently in nature for very high sociability species or decreased willingness and motivation to join and stay within shoals for very low sociability species. Notably, the numerical discrimination was sensitive to the numerical contrast ratio in LP species but not in HP species, suggesting that the numerical system used for size discrimination also varied between species. Overall, we demonstrated the interspecies relationship between personality and shoal size discrimination across fish species, suggesting an evolutionary link between numerical abilities and behavior.
Subject(s)
Choice Behavior , Fishes , Animals , Cognition , Personality , Fresh WaterABSTRACT
Limited aerobic scope (AS) during digestion might be the main constraint on the performance of bodily functions in water-breathing animals. Thus, investigating the postprandial changes in various physiological functions and determining the existence of a shared common pattern because of possible dependence on residual AS during digestion in freshwater fish species are very important in conservation physiology. All species from slow-flow habitats showed impaired swimming speed while digesting, whereas all species from fast-flow habitats showed strong swimming performance, which was unchanged while digesting. Only two species from slow-flow habitats showed impaired heat tolerance during digestion, suggesting that whether oxygen limitation is involved in the heat tolerance process is species-specific. Three species from slow- or intermediate-flow habitats showed impaired hypoxia tolerance during digestion because feeding metabolism cannot cease completely under hypoxia. Overall, there was no common pattern in postprandial changes in different physiological functions because: (1) the digestion process was suppressed under oxygen-limiting conditions, (2) the residual AS decreased during digestion, and (3) performance was related to residual AS, while digestion was context-dependent and species-specific. However, digestion generally showed a stronger effect on bodily functions in species from slow-flow habitats, whereas it showed no impairment in fishes from fast-flow habitats. Nevertheless, the postprandial change in physiological functions varies with habitat, possibly due to divergent selective pressure on such functions. More importantly, the present study suggests that a precise prediction of how freshwater fish populations will respond to global climate change needs to incorporate data from postprandial fishes.
ABSTRACT
Microplastics (< 5 mm) are widely found in organisms and have the potential harm to ecosystems. Despite their widespread prevalence in environments, there is high individual varation in the abundance of microplastics found in individuals of the same species. In the present study, juvenile cichlid fish (Chindongo demasoni) were chosen to determine the ingestion personality for microplastics in the laboratory. The visible implant fluorescent tags were used for individual recognition. The fish were fed with microplastic fiber, pellet, and food for comparison. Our results showed that the observation of the behaviors of fish could be successfully matched with subsequent measurements for each individual through the tag method in microplastic research. The difference in the abundance of fiber (0-27 items/ind.) among fish individuals was also observed in our study. Meanwhile, the abundance of fiber showed a positive correlation with the average speed and covered area of fish, which indicates the degree of activity of fish. Moreover, fish with higher speed or a front position had higher capturing times for pellet. Our results suggest that the swimming behaviors of fish affect their ingestion of microplastics, and active fish had a higher likelihood of ingesting microplastics, which might be one of the reasons for the common phenomena, i.e., great individual differences observed in microplastic studies.
Subject(s)
Cichlids , Water Pollutants, Chemical , Animals , Microplastics/toxicity , Plastics , Environmental Monitoring/methods , Ecosystem , Swimming , Water Pollutants, Chemical/toxicity , Fishes , EatingABSTRACT
Curdlan was effective in alleviating quality deterioration of frozen dough during storage. This research explored the mechanisms from perspectives of fermentation properties, water state and gluten structure of frozen dough during storage, and the performance of corresponding steamed bread. Results showed that curdlan addition improved the gas-releasing capability and gas-holding capability of frozen dough, meanwhile enhanced the specific volume and textural properties of corresponding steamed bread. The melting enthalpy and NMR results demonstrated that curdlan restricted the conversation of bound water into freezable water, and inhibited the moisture migration in frozen dough. Frozen dough with 0.5% curdlan had significantly lower gluten macropolymers (GMP) depolymerization degree and free sulfhydryl (SH) content than the control, indicating that curdlan alleviated the depolymerization of GMP. Microstructure results proved that the deterioration of the structure was retarded by curdlan. This study contributes to understanding the theories for curdlan alleviating the deterioration of frozen dough during storage.
ABSTRACT
Phenotypic plasticity has been identified as a major mechanism of response to changing temperatures. Parental effects are potentially important drivers of ecological and evolutionary dynamics, while developmental plasticity also plays a key role in generating phenotypic variation. However, little is known of the interaction between parental effects and developmental plasticity on the thermal phenotypes of fishes with different reproductive modes (i.e. oviparous vs. viviparous). To understand the contributions of inter- and intra-generational plasticity of thermal phenotypes (preferred temperature, avoidance temperatures, critical thermal thresholds) in fishes with different reproductive modes, we carried out a factorial experiment in which both breeding parents and offspring were exposed to lower (22 °C) or higher (28 °C) temperatures, using zebrafish (Danio rerio) and guppies (Poecilia reticulata) as representative oviparous and viviparous species. We found that offspring thermal preference and avoidance of both species were significantly influenced by parental effects and developmental plasticity, with higher thermal preference and avoidance consistent with higher background (parental) temperature treatments. However, parental effects were only found to impose significant effect on the thermal tolerances of guppies. The findings suggest that phenotypic plasticity, both within and across generations, may be an important mechanism to adapt to rapid climate changes, and that future temperature fluctuations may impose more profound effects on viviparous fish species in general.
ABSTRACT
Flow stimulation before release into the wild may contribute to improved survivability of farmed fish. However, the effects of flow stimulation on the survival rate of fish depend on the fish species and exercise regime, such as exercise type, duration, and intensity. In this study, juvenile Percocypris pingi swam for 18 h per day for 8 weeks under different water speeds, 3 cm s-1 (control) and 1, 2, and 4 body lengths (bl) s-1 , at 20°C. Then, parameters related to the growth rate, swimming capacity, spontaneous activity, and immune function were measured. We found that (1) continuous flow stimulation had no significant influence on the growth but was conducive to the increase in the relative carcass mass; (2) continuous flow stimulation at 2 or 4 bl s-1 enhanced the aerobic swimming capacity (Ucrit ), which may be due to an increase in anaerobic exercise capacity (endurance time) rather than to changes in maximum metabolic rate and aerobic scope; (3) continuous flow stimulation at 4 bl s-1 led to a significant increase in spontaneous activity, which was mainly due to the higher percent time spent moving as compared with the controls; and (4) continuous flow stimulation at 2 bl s-1 may contribute to improving the nonspecific immune parameter (lysozyme activity) in juvenile P. pingi. Our findings suggest that continuous flow stimulation at 2 or 4 bl s-1 for 18 h per day for 56 days at 20°C before release in wild may be a suitable training regime for improving the survival rate of cultured juvenile P. pingi.
Subject(s)
Cyprinidae , Swimming , Animals , Swimming/physiologyABSTRACT
LncRNA HOTAIR exhibited different effects in human cancers. However, the role of HOTAIR was not reported in osteosarcoma. This study aimed to explore the function of HOTAIR in osteosarcoma. Firstly, we examined HOTAIR expression in breast cancer tissues by the RT-qPCR assay and examined HOTAIR protein expression via immunocytochemistry, to chemical assay, and Western blot. Then, for further exploring the function of HOTAIR, we also examined it by CCK-8 and transwell assays. Downregulation of HOTAIR was detected in osteosarcoma, which predicted poor prognosis of patients with osteosarcoma. Moreover, cell migration, invasion, and proliferation were suppressed by HOTAIR overexpression in osteosarcoma. Furthermore, LPR5 was a direct target of HOTAIR, which was upregulated in osteosarcoma. Especially, the upregulation of LPR5 could impair the suppressive effect of HOTAIR in breast cancer. HOTAIR was found to negatively regulate the EMT and Wnt/ß-cadherin pathways in osteosarcoma. HOTAIR repressed the progression of osteosarcoma via regulating LPR5 and suppressing the Wnt/ß-cadherin pathway. Our findings will provide a positive reference for studying the function of HOTAIR in osteosarcoma.
ABSTRACT
Growth and energy metabolism are highly flexible in fish species in response to food availability, and these two traits depend to some extent on the social rearing environment (e.g., isolated vs. group rearing). Currently, how social rearing environments influence flexibility in metabolic rate of fish and their ecological consequences (e.g., somatic growth) remain largely unknown. Here, we investigated how social isolation (i.e., group-reared vs. isolation-reared) and food availability (i.e., high vs. low) affect metabolic rates, growth and their correlations in a group-living fish, grass carp (Ctenopharyngodon idella), which were subjected to a 4-week growth experiment. The metabolic rates (e.g., standard metabolic rate, SMR; maximum metabolic rate, MMR; aerobic scope, AS = MMR-SMR) and morphology (e.g., body mass and length) of the fish in four treatments were measured at the beginning and end of the growth experiment, and then the growth parameters (e.g., food intake, FI; feeding efficiency, FE; and specific growth rate, SGR) were also obtained. We found that social isolation did impair growth of fish with individuals showing a lower SGR compared to those group-reared fish irrespective of food availability. However, the growth advantage of group-reared fish under two food availabilities did not result from their FIs or FEs. Metabolic rates (i.e., SMR) seemed to decrease in response to social isolation, but increased greater when fish were reared at high food ration. These shifts in metabolic rates were positively linked with individual differences in somatic growth; individuals who increased metabolic rates more grew faster, while those who increased their metabolic rates less or even reduced had a lower growth, but these links were independent on both social isolation and food ration. These results suggested that social isolation can inhibit the growth of individual fish, but not the AS. Flexibility in metabolic rates could confer a growth advantage under changing food availability, but the links between variation in energy metabolism and growth were not altered by social deprivation. Our study demonstrates the importance of metabolic plasticity accounting for inter-individual difference in growth performance under the challenges of changing food resource.
Subject(s)
Carps , Animals , Energy Metabolism/physiology , Food , Social IsolationABSTRACT
Exercise and aerobic scope in fishes have attracted scientists' attention for several decades. While it has been suggested that aerobic scope may limit behavioral expression and tolerance to environmental stressors in fishes, the exact importance of aerobic scope in an ecological context remains poorly understood. In this review, we examine the ecological relevance of aerobic scope by reconsidering and reanalyzing the existing literature on Chinese freshwater fishes across a wide-range of habitats and lifestyles. The available evidence suggests that natural selection in fast-flowing aquatic habitats may favor species with a high aerobic scope and anaerobic capacity for locomotion, whereas in relatively slow-flowing habitats, hypoxia tolerance may be favored at the cost of reduced locomotor capacity. In addition, while physical activity can usually cause fishes from fast-flowing habitats to reach their aerobic metabolic ceiling (i.e., maximum metabolic rate), possibly due to selection pressure on locomotion, most species from slow-flowing habitats can only reach their metabolic ceiling during digestion, either alone or in combination with physical activity. Overall, we suggest that fish exhibit a continuum of metabolic types, from a 'visceral metabolic type' with a higher digestive performance to a 'locomotion metabolic type' which appears to have reduced capacity for digestion but enhanced locomotor performance. Generally, locomotor-type species can either satisfy the demands of their high swimming capacity with a high oxygen uptake capacity or sacrifice digestion while swimming. In contrast, most visceral-type species show a pronounced decrease in swimming performance while digesting, probably owing to conflicts within their aerobic scope. In conclusion, the ecological relevance of aerobic scope and the consequent effects on other physiological functions are closely related to habitat and the lifestyle of a given species. These results suggest that swimming performance, digestion and hypoxia tolerance might coevolve due to dependence on metabolic traits such as aerobic scope.
Subject(s)
Fishes , Swimming , Animals , Digestion , Ecosystem , Hypoxia , Swimming/physiologyABSTRACT
Learning and memory abilities and their roles in group decision-making have important ecological relevance in routine activities such as foraging and anti-predator behaviors in fish species. The aims of the present study were to explore individual spatial learning abilities of juvenile cichlids (Chindongo demasoni) in a foraging context, and to explore the influence of heterogeneity of memory information among group members on group performance in a six-arm radiation maze. In the context of an association between landmarks and food, learning ability was evaluated by the speed and accuracy of reaching the arm with food during seven days of reinforcement, and memory retention was tested at intervals of 2, 5, 8 and 11 days of detraining. Then, the speed and accuracy of an eight-member group with different proportions of memory-trained fish were measured. Both speed and accuracy of individual fish improved significantly and linearly in the first five days of training and leveled off between five and seven days, with values 60% shorter (in speed) and 50% higher (in accuracy) compared to those of the first day. Neither speed nor accuracy showed any decrease after 11 days of detraining, suggesting memory retention of the spatial task. When measured in a group, the speed and accuracy of the majority of the group (more than half) in reaching the arm with food changed linearly with an increasing ratio of trained members. This shows that cichlids can acquire associative learning information through a training process, and group behavior of cichlids seems not likely be determined by a minority of group members under a foraging context.
ABSTRACT
The Chinese giant salamander (Andrias davidianus), one of the largest extant amphibian species, has dramatically declined in the wild. As an ectotherm, it may be further threatened by climate change. Therefore, understanding the thermal physiology of this species should be the priority to formulate related conservation strategies. In this study, the plasticity in metabolic rate and thermal tolerance limits of A. davidianus larvae were studied. Specifically, the larvae were acclimated to three temperature levels (7 °C, cold stress; 15 °C, optimum; and 25 °C, heat stress) and two diet items (red worm or fish fray) for 20 days. Our results indicated that cold-acclimated larvae showed increased metabolic capacity, while warm-acclimated larvae showed a decrease in metabolic capacity. These results suggested the existence of thermal compensation. Moreover, the thermal tolerance windows of cold-acclimated and warm-acclimated larvae shifted to cooler and hotter ranges, respectively. Metabolic capacity is not affected by diet but fish-fed larvae showed superiority in both cold and heat tolerance, potentially due to the input of greater nutrient loads. Overall, our results suggested a plastic thermal tolerance of A. davidianus in response to temperature and diet variations. These results are meaningful in guiding the conservation of this species.
ABSTRACT
Many studies have found that aerobic exercise training at a moderate water velocity can improve the growth, swimming performance and survival rate of fish. To investigate the effects of aerobic exercise training on the growth, swimming performance, antipredation ability and immune parameters of rock carp, juveniles were placed in training channels with different water velocities (i.e., 3 cm s-1, 1 (body length s-1) bl s-1, 2 bl s-1 and 4 bl s-1) for 6 weeks. Then, the specific growth rate, critical swimming speed (Ucrit) and its metabolism, constant acceleration speed (Ucat), survival rate under predation, spleen index, lysozyme (LZM) activity and immunoglobulin (IgM) level were measured. Training showed no significant effect on the length-specific growth rate, weight-specific growth rate, Ucrit, maximum metabolic rate (MMR), metabolic scope (MS), Ucat or spleen index. The resting metabolic rates (RMRs) of the 2 bl s-1 and 4 bl s-1 training groups were significantly higher than those of the control group and 1 bl s-1 training group. The survival rate of the 1 bl s-1 training group in the presence of predators was significantly higher than that of the control group but significantly lower than those of the 2 bl s-1 and 4 bl s-1 training groups. The LZM activity of the 4 bl s-1 training group was significantly higher than that of the control group. The IgM level of the 2 bl s-1 training group was significantly higher than that of the control group. These data indicate that aerobic exercise training does not improve the growth and swimming performance of juvenile rock carp but can improve their antipredation ability and immunologic function.
ABSTRACT
Physicochemical properties and chemical composition of Chinese perilla seed oil has been characterized in this study. The result showed that both the cold press oil and the solvent extracted oil possessed low acid value and peroxide value. The fatty acid composition result showed that the oil has high content of linolenic acid (C18:3) up to 66.4 g/100 g, followed by linoleic acid (C18:2) of 15.3 g/100 g. The total triacylglycerol (TAG) profiles results showed that the oil contained 20 TAGs including 17 regioisomers, including LnLnLn (35.8 g/100 g), LLnLn (20.2 g/100 g), LLLn (17.7 g/100 g) and PLnLn (14.9 g/100 g) (Ln, linolenic acid; L, linoleic acid; P, palmitic acid). With content of only 0.57 g/100 g oil, the unsaponifiable matters were mainly composed of phytosterols, squalene, tocopherol, alcohols and hydrocarbons. The total phytosterols content was 0.39 g/100 g oil, in which ß-sitosterol has high content of 0.31 g/100 g oil.
Subject(s)
Chemical Phenomena , Linoleic Acid/analysis , Perilla frutescens/chemistry , Phytosterols/analysis , alpha-Linolenic Acid/analysis , Alcohols/analysis , Antioxidants/analysis , Hydrocarbons/analysis , Isomerism , Liquid-Liquid Extraction/methods , Palmitic Acid/analysis , Plant Oils/chemistry , Plant Oils/isolation & purification , Squalene/analysis , Tocopherols/analysis , Triglycerides/analysis , alpha-Linolenic Acid/chemistry , alpha-Linolenic Acid/isolation & purificationABSTRACT
The present study aimed to reveal the effect of fasting (21 days) on collective movement and interaction dynamics in both homogeneous (eight members fed a commercial diet or deprived of food) and heterogeneous (four fed + four starved members) shoals of juvenile qingbo (Spinibarbus sinensis). The authors of this study measured the shoaling behaviour in both a commonly used rectangular open arena with no spatial complexity and a radial arm maze. When measured in the open arena, the starved shoals had a faster swimming speed and acceleration rate and a longer interindividual distance than the fed shoals, possibly because of the elevated foraging motivation. Nonetheless, the values of the heterogeneous groups were similar to those of the fed groups. Furthermore, in contrast to the fish in homogeneous shoals, the starved fish in heterogeneous shoals showed a slower acceleration rate and speed than fed members in heterogeneous shoals. These results, combined with the relationships of variables at the among- and within-shoal levels, suggested that starved fish limited their motion in heterogeneous shoals to maintain group cohesion but that the fed fish contributed more to maintaining shoal structure, possibly because of the higher energy expenditure required for movement changes. When monitored in a radial arm maze, starved shoals showed more fission-fusion episodes without sacrificing group cohesion, as they adaptively adjusted the frequency and duration of each majority choice. The among-shoal variation revealed that the heterogeneous groups showed less variation in the open arena but more variation in the radius maze than did the homogeneous groups. This difference might arise because dominant members have opposite effects on shoal behaviour and consensus decisions. In conclusion, the present study showed opposite effects of feeding states on collective behaviour between homogeneous and heterogeneous shoals, possibly because of the complicated interactions among members with different energy storage levels and foraging motivations. Furthermore, the heterogeneous groups showed a difference between shoal behaviour in the open area and exploration in the radial arm maze. Future studies manipulating the personality composition of starved and fed members of heterogeneous groups might yield interesting results.
Subject(s)
Cyprinidae , Social Behavior , Animals , Fasting , SwimmingABSTRACT
Almost all prey live in habitats with predators with different hunting modes; however, most studies on predation have investigated the effects of only one predator at a time. In this study, we aimed to investigate whether qingbo (Spinibarbus sinensis), a common cyprinid fish, responds differently to active hunting and ambush predators and how qingbo responds when both types of predators coexist. Juvenile qingbo were subjected to catfish (Clarias fuscus, active hunter) exposure, snakehead fish (Channa argus, ambush hunter) exposure, or mixed predator exposure (catfish and snakehead coexistence) for a duration of 60 days. Then, their growth, behaviors, swimming performance, and metabolism were measured. Qingbo subjected to active hunting predator exposure exhibited decreased activity and predator inspection and improved fast-start escape performance compared to those in the control group. However, none of the parameters of the fish subjected to ambush predator exposure changed significantly. Fish subjected to mixed predator exposure exhibited improved fast-start escape performance but increased maintenance energy expenditure, whereas no changes were observed in any of the behavioral variables. Qingbo showed a stronger anti-predator response to active hunting predators than to ambush predators, suggesting that the fish exhibit a stronger anti-predator response to a current direct threat than to a potential threat (a predator exists nearby but seldom presents in attack behavior). Additionally, the response of prey fish to multiple predators was quite complex, and the coexistence and interaction of multiple predator species with different hunting modes may lead to serious stress responses and confound the prey's behavioral responses to each predator.