Overview on the role of dietary Spirulina platensis on immune responses against Edwardsiellosis among Oreochromis niloticus fish farms.

Edwardsiellosis is a bacterial fish disease that mostly occurs in freshwater farms and is characterized by a high mortality rate. Edwardsiella tarda strain was recovered from 17 fish out of 50 Nile tilapia, which were harboring clinical signs of systemic septicemia. The level of un-ionized ammonia (NH3) in the fish farm's water was 0.11-0.15 mg/L, which was stressful for the Nile tilapia.Sequencing of the gyrB1 gene confirmed that the isolate was E. tarda JALO4, and it was submitted to NCBI under the accession number PP449014. The isolated E. tarda harbored the virulence gene edw1 AHL-synthase (quorum sensing). In addition, the isolate was sensitive to trimethoprim and sulfamethoxazole mean while it was intermediate to florfenicol. The median lethal dose (LD50) of E. tarda JALO4 was determined to be 1.7 × 105 CFU/mL in Nile tilapia.In the indoor experiment, Nile tilapia (45.05 ± 0.4 g), which received dietary Spirulina platensis (5 and 10 g/kg fish feed), showed optimum growth and feed utilization. Meanwhile, after receiving dietary S. platensis, the fish's feed conversion ratio (FCR) was significantly enhanced compared to the control, which was 1.94, 1.99, and 2.88, respectively. The expression of immune-related genes interleukin (IL)-1ß and tumor necrosis factor (TNF)-α were upsurged in E. tarda-challenged fish with higher intensity in S. platensis groups. Dietary S. platensis at a dose of 10 g/kg fish feed could provide a relative protection level (RPL) of 22.2% Nile tilapia challenged against E. tarda. Nile tilapia experimentally infected E. tarda, drastically altering their behavior: higher operculum movement, low food apprehension, and abnormal swimming dietary S. platensis (10 g/kg fish feed) could rapidly restore normal status.It was concluded that Edwardsiellosis could alter Nile tilapia behavior with a high loss in fish population. Fish received dietary-S. platensis could rapidly restore normal behavior after E. tarda infection. It is recommended the incorporation of S. platensis at doses of 10 g/kg into the Nile tilapia diet to boost their immunity and counteract E. tarda infection.

Shy and Bold Fish Have the Same Preference for Light Color Selection.

Personality, which matters for animal welfare, demonstrates behavioral differences. Light is one of the most important factors in aquaculture. However, how fish personality affects light color selection is unclear. In this study, we tested the personality of yellow catfish Pelteobagrus fulvidraco juveniles and then quantified the selective behaviors of different personalities under six light colors: violet (410-420 nm), yellow (580-590 nm), green (550-560 nm), red (620-630 nm), blue (470-480 nm), and white. The results showed that juveniles preferred the yellow and green light over the other colors of light, probably due to different reasons. The average cumulative dwell time in yellow (32.81 ± 5.22%), green (21.81 ± 3.58%), and red (26.36 ± 4.89%) lights was significantly longer than the other light colors, and the average visit frequency in green light (32.00 ± 4.93%) was the most. Juveniles had the longest total moved distance in green light. Moreover, the results demonstrated that shy and bold individuals had the same preference for the green light. Bold individuals could find the preferred light colors rapidly and make quick decisions for light color selection. After identifying the preferred light colors, bold individuals reduced the frequency of exploration. This study provides a theoretical basis for the welfare of juvenile yellow catfish in aquaculture.

Archerfish foraging success varies with immediate competition level but not group size.

Group living can lead to kleptoparasitism, the theft of resources by competitors. Under such conditions, foragers may alter their behavior to minimize competition. However, it is unclear how such behavioral changes impact foraging performance. Archerfish (Toxotes spp.) are a good model for investigating the behavioral responses to kleptoparasitism, as their hunting method (shooting waterjets at insects perched above the water) leaves them vulnerable to theft. They must hit the target prey with sufficient force to dislodge it; thus, the prey may land some distance away from the shooter. Kleptoparasitism rates increase with group size in archerfish, and individuals alter their behavior around conspecifics. We investigated whether group size affected shooting success, using 7-spot archerfish T. chatareus. We considered a fish's shot to be successful if it knocked a fly, placed on a transparent platform above the tank, into the water. The probability of shooting success was modeled as a function of group size, aiming duration, nearest neighbor distance and position, and trial number. We found no effect of group size, aiming duration, or nearest neighbor distance or position on shooting success. Shooting success increased as trials progressed, likely due to the fish becoming more familiar with the task. We also found no change in the kleptoparasitism rate between group sizes. Instead, the likelihood of the shooter consuming the prey depended on the types of competition present at the time of shooting. We suggest that archerfish shooting behavior can be influenced by the presence of conspecifics in ways not previously considered.

Quantitative comparison of 2D and 3D monitoring dimensions in fish behavior analysis.

To improve the accuracy and efficiency of fish behavior assessment, this paper focuses on quantitatively exploring the variations and relationships between different monitoring dimensions. A systematic comparison was conducted between 3D and 2D behavioral factors using an infrared tracing system, during both day and night. Significant differences in swimming distance were observed among the different monitoring methods, as determined by two-way ANOVA and Tukey's test. A correction was applied to account for the disparities observed in 2D swimming distance, ensuring accurate measurements. These findings present a cost-effective and efficient approach for obtaining precise 3D distance data. Additionally, a kinematic factor called the "number of U-turns" was proposed to provide a more intuitive characterization of directional changes in fish swimming. Significant differences were observed between 2D and 3D data, with higher percentages of false U-turn counts and missing U-turn counts compared to correct counts in the 2D view. These findings suggest that reducing the monitoring dimension may impact the accurate estimation of swimming motion, potentially resulting in inaccurate outcomes. Finally, the statistical analyses of the non-linear properties of fractal dimension revealed significant differences among the various monitoring methods. This conclusion has practical implications for biologists and physicists, enabling them to improve the accuracy of behavioral phenotyping for organisms exhibiting 3D motion.

Differences in predator-avoidance behavior between two invasive gobies and their native competitors.

Globally, fish are frequently introduced beyond their native range. Some, like Ponto-Caspian gobies, are becoming invasive, achieving high colonization rates and constituting frequent prey for native predators. However, little is known about the effectiveness of antipredator behaviors of the invaders, which may shape their role in the invaded community and contribute to the invasion success. We compared antipredator behaviors of invasive gobies and native fish species after their detection by the predator, when the danger becomes direct. We studied 2 fish pairs, each consisting of an invasive and native species co-occurring in the environment and belonging to the same prey guild: (1) the racer goby Babka gymnotrachelus versus European bullhead Cottus gobio, (2) the monkey goby Neogobius fluviatilis versus gudgeon Gobio gobio, facing a naïve predator (the Eurasian perch Perca fluviatilis). We analyzed behaviors of single prey individuals (escaping, staying in shelter, and activity) and single predators (activity, searching, following, capturing, and latency to prey consumption). In the predator presence, the bullhead was less active and more often managed to escape after capture than the racer goby. The gudgeon escaped before the capture more often than the monkey goby. The predator succeeded later with the bullhead compared to racer goby, whereas no differences in ingestion time occurred between the gudgeon and monkey goby. The results suggest that, in terms of hunting effort of native predators, the invasive gobies are equivalent to or more profitable prey than their native analogs, which can facilitate the integration of the gobies into local food webs.

What acoustic telemetry can and cannot tell us about fish biology.

Acoustic telemetry (AT) has become ubiquitous in aquatic monitoring and fish biology, conservation, and management. Since the early use of active ultrasonic tracking that required researchers to follow at a distance their species of interest, the field has diversified considerably, with exciting advances in both hydrophone and transmitter technology. Once a highly specialized methodology, however, AT is fast becoming a generalist tool for those wishing to study or conserve fishes, leading to diversifying application by non-specialists. With this transition in mind, we evaluate exactly what AT has become useful for, discussing how the technological and analytical advances around AT can address important questions within fish biology. In doing so, we highlight the key ecological and applied research areas where AT continues to reveal crucial new insights and, in particular, when combined with complimentary research approaches. We provide a comprehensive breakdown of the state of the art for applications of AT, discussing the ongoing challenges, where its strengths lie, and how future developments may revolutionize fisheries management, behavioral ecology and species protection. Through selected papers we illustrate specific applications across the broad spectrum of fish biology. By bringing together the recent and future developments in this field under categories designed to broadly capture many aspects of fish biology, we hope to offer a useful guide for the non-specialist practitioner as they attempt to navigate the dizzying array of considerations and ongoing developments within this diverse toolkit.

Rhythm of relationships in a social fish over the course of a full year in the wild.

BACKGROUND: Animals are expected to adjust their social behaviour to cope with challenges in their environment. Therefore, for fish populations in temperate regions with seasonal and daily environmental oscillations, characteristic rhythms of social relationships should be pronounced. To date, most research concerning fish social networks and biorhythms has occurred in artificial laboratory environments or over confined temporal scales of days to weeks. Little is known about the social networks of wild, freely roaming fish, including how seasonal and diurnal rhythms modulate social networks over the course of a full year. The advent of high-resolution acoustic telemetry enables us to quantify detailed social interactions in the wild over time-scales sufficient to examine seasonal rhythms at whole-ecosystems scales. Our objective was to explore the rhythms of social interactions in a social fish population at various time-scales over one full year in the wild by examining high-resolution snapshots of a dynamic social network. METHODS: To that end, we tracked the behaviour of 36 adult common carp, Cyprinus carpio, in a 25 ha lake and constructed temporal social networks among individuals across various time-scales, where social interactions were defined by proximity. We compared the network structure to a temporally shuffled null model to examine the importance of social attraction, and checked for persistent characteristic groups over time. RESULTS: The clustering within the carp social network tended to be more pronounced during daytime than nighttime throughout the year. Social attraction, particularly during daytime, was a key driver for interactions. Shoaling behavior substantially increased during daytime in the wintertime, whereas in summer carp interacted less frequently, but the interaction duration increased. Therefore, smaller, characteristic groups were more common in the summer months and during nighttime, where the social memory of carp lasted up to two weeks. CONCLUSIONS: We conclude that social relationships of carp change diurnally and seasonally. These patterns were likely driven by predator avoidance, seasonal shifts in lake temperature, visibility, forage availability and the presence of anoxic zones. The techniques we employed can be applied generally to high-resolution biotelemetry data to reveal social structures across other fish species at ecologically realistic scales.

Enhancing Rearing of European Seabass (Dicentrarchus labrax) in Aquaponic Systems: Investigating the Effects of Enriched Black Soldier Fly (Hermetia illucens) Prepupae Meal on Fish Welfare and Quality Traits.

Within the modern aquaculture goals, the present study aimed to couple sustainable aquafeed formulation and culturing systems. Two experimental diets characterized by 3 and 20% of fish meal replacement with full-fat spirulina-enriched black soldier fly (Hermetia illucens) prepupae meal (HPM3 and HPM20, respectively) were tested on European seabass (Dicentrarchus labrax) juveniles during a 90-day feeding trial performed in aquaponic systems. The experimental diets ensured 100% survival and proper zootechnical performance. No behavioral alterations were evidenced in fish. Histological and molecular analyses did not reveal structural alterations and signs of inflammation at the intestinal level, highlighting the beneficial role on gut health of bioactive molecules typical of HPM or derived from the enriching procedure of insects' growth substrate with spirulina. Considering the quality traits, the tested experimental diets did not negatively alter the fillet's fatty acid profile and did not compromise the fillet's physical features. In addition, the results highlighted a possible role of spirulina-enriched HPM in preserving the fillet from lipid oxidation. Taken together, these results corroborate the use of sustainable ingredients (spirulina-enriched HPM) in aquaponic systems for euryhaline fish rearing.

Behavioral, biochemical, and endocrine responses of zebrafish to 30-min exposure with environmentally relevant concentrations of imidacloprid-based insecticide.

The imidacloprid-based insecticides (IBIs) are among the most used insecticides worldwide, and chronic and acute toxic effects (days exposure protocols) have been reported in several species in studies of IBIs at lethal concentrations. However, there is little information on shorter time exposures and environmentally relevant concentrations. In this study, we investigated the effect of a 30-min exposure to environmentally relevant concentrations of IBI on the behavior, redox status, and cortisol levels of zebrafish. We showed that the IBI decreased fish locomotion and social and aggressive behaviors and induced an anxiolytic-like behavior. Furthermore, IBI increased cortisol levels and protein carbonylation and decreased nitric oxide levels. These changes were mostly observed at 0.013 and 0.0013 µg·L-1 of IBI. In an environmental context, these behavioral and physiological disbalances, which were immediately triggered by IBI, can impair the ability of fish to evade predators and, consequently, affect their survival.

Infection of Atlantic tripletail Lobotes surinamensis (Teleostei: Lobotidae) by brain metacercariae Cardiocephaloides medioconiger (Digenea: Strigeidae).

Three juvenile Atlantic tripletail Lobotes surinamensis caught opportunistically in Charleston Harbor (South Carolina, USA) and maintained in captivity for over three months displayed an altered swimming behavior. While no direct causation can be demonstrated herein, fish were infected in their brain by strigeid trematode larvae (metacercariae) of Cardiocephaloides medioconiger, which were identified via ITS2 and 28S ribosomal RNA gene sequencing. Histology showed nonencysted metacercariae within the brain ventricle between the optic tectum and tegmentum, causing distortion of tegmental parenchyma. Aggregates of mononuclear inflammatory cells were in the ventricle adjacent to metacercariae. Metacercarial infection by Cardiocephaloides medioconiger has been reported from the brain and eyes of only two other fish species from the northern US Atlantic coast: the grey mullet Mugil cephalus and silverside Menidia menidia, but this identification is problematic and needs molecular verification. Atlantic tripletail is a new report as a second intermediate host for C. medioconiger and South Carolina is a new locality. Cardiocephaloides species in general have a low host specificity and infection by C. medioconiger could propagate to other fishes and affect neighboring natural ecosystems.

Sorbed environmental contaminants increase the harmful effects of microplastics in adult zebrafish, Danio rerio.

Aquatic animals ingest Microplastics (MPs) which have the potential to affect the uptake and bioavailability of sorbed co-contaminants. However, the effects on living organisms still need to be properly understood. The present study was designed to assess the combined effects of MPs and environmental contaminants on zebrafish (Danio rerio) health and behavior. Adult specimens were fed according to three different protocols: 1) untreated food (Control group); 2) food supplemented with 0.4 mg/L pristine polyethylene-MPs (PE-MPs; 0.1-0.3 mm diameter) (PEv group); 3) food supplemented with 0.4 mg/L PE-MPs previously incubated (PEi group) for 2 months in seawater. Analysis of contaminants in PEi detected trace elements, such as lead and copper. After 15 days of exposure, zebrafish underwent behavioral analysis and were then dissected to sample gills and intestine for histology, and the latter also for microbiome analysis. Occurrence of PEv and PEi in the intestine and contaminants in the fish carcass were analyzed. Both PEv- and PEi-administered fish differed from controls in the assays performed, but PEi produced more harmful effects in most instances. Overall, MPs after environmental exposure revealed higher potential to alter fish health through combined effects (e.g. proportion of microplastics, pollutants and/or microorganisms).

Chronic poly(l-lactide) (PLA)- microplastic ingestion affects social behavior of juvenile European perch (Perca fluviatilis).

Juvenile perch were exposed to 2 % (w/w) poly(l-lactide) (PLA) microplastic particles (90-150 µm) in food pellets, or 2 % (w/w) kaolin particles, and a non-particle control food over 6 months. Chronic ingestion of PLA microplastics significantly affected the social behavior of juvenile perch, evident as a significantly increased reaction to the vision of conspecifics. PLA ingestion did not alter life cycle parameters, or gene expression levels. In addition to reactions to conspecifics, fish that ingested microplastic particles showed tendencies to decrease locomotion, internal schooling distance, and active predator responses. The ingestion of natural particles (kaolin) significantly downregulated the expression of genes related to oxidative stress and androgenesis in the liver of juvenile perch, and we found tendencies to downregulated expression of genes related to xenobiotic response, inflammatory response, and thyroid disruption. The present study demonstrated the importance of natural particle inclusion and the potential behavioral toxicity of one of the commercially available biobased and biodegradable polymers.

Feeding habits of four-finger threadfin fish, Eleutheronema tetradactylum, and its diet interaction with co-existing fish species in the coastal waters of Thailand.

This study assessed the feeding habits of four-finger threadfin fish, Eleutheronema tetradactylum, and its diet relationship with other fish species in the tropics. Fish samples were collected from four locations along the coastal regions of Thailand. A whole year field sampling event was conducted to investigate the diet relationship of threadfin fish with other ten co-existing fish species in Pattani Bay during January 2021 and January 2022. E. tetradactylum was an active and specific predator with significant diet shift during ontogeny. Specifically, the juvenile fish fed largely on zooplankton especially Acetes/shrimp postlarvae, and small sized-fish fed on penaeid shrimps, while medium and large-sized fish shifted their diets to a combination of penaeid shrimp, fish and squid. Size and sex of fish as well as site of collection significantly affected gut fullness index and average number of food type (p < 0.05). Transitional sex fish predated almost entirely on other fishes (87.2%), whereas male and female fish fed mainly on penaeid shrimp (66.5%) and other fish (51.3%), respectively. Fish size and mouth opening controlled the size of prey, with the larger fish with larger mouth-opening fed primarily on the larger size of prey. Moreover, E. tetradactylum shared its diets inclusively with Epinephelus coioides, Johnius belangerii, Scomberomorus commerson, Scomberoides lysan, Otolithes ruber and Lutjanus russelli. Penaeid shrimp and teleost fish were the main food types shared by these fishes. This study provided important information on the feeding habits of E. tetradactylum and its diet relationship with other co-existing fish species living in the same habitat of a tropical coastal region.

Overwintering aggregation patterns of European catfish Silurus glanis.

Animal aggregation, particularly in large-bodied species, is both a fascinating and intriguing phenomenon. Here we analyzed the overwintering behavior of the European catfish, Silurus glanis Linnaeus, 1758, the largest freshwater fish in Europe. By tracking 47 subadults and adults in a shallow lake in southeastern France, we reported a consistent aggregative behavior across four successive winters. By implementing time series analysis and Cox proportional hazard models, we investigated the dynamics of these aggregations (formation, stability, dislocation), and the factors that govern it, whether external (temperature, time of the day) or specific to the fish (size, key individuals). These aggregations lasted 1.5-2 months and mainly took place in a single small 4 m-deep area whose environmental conditions (temperature, oxygen, substrate) did not differ from other parts of the lake. In some periods during winter, all tagged fish were aggregated, which suggests that a large proportion of the lake population gathered there. Low temperatures (below 9 °C) triggered the formation of aggregations. They became more stable with decreasing temperatures, while individuals more frequently left the aggregation, preferentially at dusk and at night, when temperatures increased. The largest individuals swam more frequently back and forth to the aggregation. Irrespective of their size, some individuals consistently arrived earlier in the aggregation in winter and left later. This predictable seasonal grouping of individuals and, more generally, the knowledge provided by such studies on how species use space have important operational value and are useful for species conservation as well as for species control.

Combined effects of high temperature and pesticide mixture exposure on free-swimming behaviors and hepatic cytochrome P450 1A expression in goldfish, Carassius auratus.

The synergy between multiple compounds and other stressors, including heat, creates volatility and greater unpredictability than standard single-chemical toxicity testing, especially in the case of pesticides and metabolites which might contain several noxious ingredients resulting in adverse ecological effects. To address this, the aim of this study was to examine the dose- and time-dependent effects of low- and high-dose pesticide mixture (metalachlor, linuron, isoproturon, tebucanazole, aclonifen, atrazine, pendimethalin, azinphos-methyl) and heat stress co-exposure (22°C control/32°C treatment for 4-week) on free-swimming behaviors and cumulative actionless time (CAT) of goldfish. Behavioral analysis showed a dose- and time-dependent decrease in distance swam, as well as a subsequent increase in CAT. Vertical and horizontal spatial behavioral use were affected under heat and pesticides co-exposure conditions. In 3- and 4-week(s) exposure groups, horizontal spatial behavioral use demonstrated elevated time spent in the lower third of the aquarium. Similarly, during 3- and 4-week(s) exposure (32°C control and 32°C high doses) vertical spatial behavioral use was found to increase time spent in the outermost edges of the aquarium. In all treatment groups, the final condition factor (KM) showed significant attenuation when compared to the initial KM. However, there was an unclear relationship between heat/pesticide co-exposure and growth most notably in 32°C high-dose groups. In addition, the expression of hepatic cytochrome P450 1A mRNA was significantly higher in pesticide-exposed groups. Taken together, data demonstrated that co-exposure with low- or high-dose pesticide mixture and heat stress significantly impacted natural swimming patterns, which over time might result in the broader population and ecological effects.

Eco-evolutionary dynamics of Atlantic cod spatial behavior maintained after the implementation of a marine reserve.

The effects of marine reserves on the life history and demography of the protected populations are well-established, typically increasing population density and body size. However, little is known about how marine reserves may alter the behavior of the populations that are the target of protection. In theory, marine reserves can relax selection on spatial behavioral phenotypes that were previously targeted by the fishery and also drive selection in favor of less mobile individuals. In this study, we used acoustic telemetry to monitor the individual spatial behavior of 566 Atlantic cod (Gadus morhua Linnaeus, 1758) moving within a marine reserve and a control site in southern Norway, starting 1 year before the implementation of the marine reserve and lasting up to 9 years after. Following a before-after-control-impact approach, we investigated changes in (1) survival, (2) selection acting on behavioral traits, and (3) mean behavioral phenotypes, after the implementation of the marine reserve. We focused on three behavioral traits commonly used to describe the mobility of aquatic animals: home range size, depth position, and diel vertical migration range. Survival increased after reserve implementation, but contrary to our expectations, it subsequently decreased to preprotection levels after just 3 years. Further, we found no significance in selection patterns acting on any of the three behavioral traits after reserve implementation. Although some changes related to water column use (the tendency to occupy deeper waters) were observed in the marine reserve after 9 years, they cannot unequivocally be attributed to protection. Our results show that survival and behavioral responses to marine reserves in some cases may be more complex than previously anticipated and highlight the need for appropriately scaled management experiments and more integrated approaches to understand the effects of marine protected areas on harvested aquatic species.

Gills Just Want to Have Fun: Can Fish Play Games, Just like Us?

It is common to observe play in dogs, cats, and birds, but have we been ignoring play in one of the most common house pets of all… fish? Aquarium fish are often used as meditative decoration in family households, but it could be that fish have similarly diverse behavioral repertoires as mammals and birds. To examine this theory, we conducted field tests at local pet stores where a range of aquarium fish species was tested for responsiveness to laser pointer stimuli. Out of 66 species of fish tested, over 80% showed a tendency to be interested in the moving laser spots, particularly red ones. Whether this behavior constitutes play is an active topic of investigation that we examine in this work.

Evaluation of Volitional Swimming Behavior of Schizothorax prenanti Using an Open-Channel Flume with Spatially Heterogeneous Turbulent Flow.

Effective fishway design requires knowledge of fish swimming behavior in streams and channels. Appropriate tests with near-natural flow conditions are required to assess the interaction between fish behavior and turbulent flows. In this study, the volitional swimming behavior of S. prenanti was tested and quantified in an open-channel flume with three (low, moderate, and high) flow regimes. The results showed that, when confronted with alternative flow regimes, S. prenanti preferred to select regions with low flow velocities (0.25−0.50 m/s) and turbulent kinetic energy (<0.05 m2/s2) for swimming, while avoiding high-turbulence areas. Moreover, S. prenanti primarily employed steady swimming behavior to search for flow velocities lower than the average current to conserve energy in low- and moderate-flow regimes. It is hypothesized that in regions with higher flow velocities, fish may change their swimming strategy from energy conservation to time conservation. Additionally, the average and maximum burst speeds of S. prenanti were 2.63 ± 0.37 and 3.49 m/s, respectively, which were 2.21- and 2.28-fold higher than the average (1.19 m/s) and maximum (1.53 m/s) burst speeds estimated from the enclosed swim chamber for fish of similar length. This study contributes a novel research approach that provides more reliable information about fish volitional swimming behavior in natural habitats, as well as recommendations for hydraulic criteria for fishways and the identification of barriers to fish migrations.

Chlorpyrifos and dichlorvos in combined exposure reveals antagonistic interaction to the freshwater fish Mrigal, Cirrhinus mrigala.

Toxicity imposed by organophosphate pesticides to the freshwater cultivable fish species mrigal (Cirrhinus mrigala) was assessed under laboratory conditions. Healthy juveniles were exposed to chlorpyrifos, dichlorvos, and their equitoxic mixture in geometric series. Median lethal concentrations of chlorpyrifos were found to be 0.906 (0.689-1.179), 0.527 (0.433-0.633), 0.435 (0.366-0.517) and 0.380 (0.319-0.450) mg/L and dichlorvos were found to be 38.432 (33.625-47.866), 22.477 (19.047-26.646), 12.442 (9.619-14.196) and 11.367 (9.496-13.536) mg/L after 24 h, 48 h, 72 h and 96 h of exposure respectively. Surprisingly, the joint toxicity of these organophosphates in the binary mixture was less than additive during most of the exposure periods. Behavioral changes exhibited by individual as well as mixture pesticide treatments were loss of schooling behavior, aggregating at corners of the test chamber, elevated opercular beatings, surplus mucus secretion, slight color changes and sudden and rapid body movements before death. Loss of fish equilibrium was noticed only in chlorpyrifos treated fish, whereas sluggish behavior was noticed only in mixture pesticide treatment. Such behavioral studies can be applied as a non-invasive bio-monitoring tool for water quality assessment for fish growth and development. Despite the same mode of action of both pesticides, the antagonistic action in the binary mixture is an interesting outcome of this research that requires further investigation for a lucid understanding of the joint toxicity mechanism of such pesticides.