Haematological assessment of the effects of the anaesthetic MS 222 in natural and neutralized form in three freshwater fish species: interspecies differences

Interspecies haematological differences to MS 222 and neutralized MS 222 anaesthesia were investigated in Sarotherodon mossambicus, Cyprinus carpio and Salmo gairdneri acclimated under identical laboratory conditions. Anaesthesia with MS 222 resulted in a 'chemical stress' in all fish, as was evident from changes in the haematological profiles of the animals. Such species specific variations in the haematology persisted throughout the whole experiment protocol which employed different concentrations of the anaesthetic. The use of neutralized MS 222, whereby aquarium water quality remained unchanged, improved the haematological profile. Possible reasons for the interspecies differences observed are discussed.     

Haematological assessment of the effects of the anaesthetic MS 222 in natural and neutralized form in three fresh water fish species: haemoglobin electrophoresis, ATP levels and corpuscular fragility curves

The effects of MS 222 and neutralized MS 222 anaesthesia on haemoglobin electrophoresis, erythrocyte adenosine triphosphate (ATP) levels and corpuscular fragility curves were studied in Cyprinus carpio, Sarotherodon mossambicus and Salmo gairdneri . Haemoglobin electrophoresis showed no significant intra-species differences in the percentage composition of the various fractions for any concentration of MS 222 and neutralized MS 222 used. Significant interspecies differences were, however, still observed. ATP levels showed intra and interspecies differences ascribed to the response of the fish species to MS 222-induced stress and not to actual changes in erythrocyte ATP concentrations. Differences were also observed in corpuscular fragility curves for all three species when using MS 222 or neutralized MS 222 compared to curves obtained without the use of the anaesthetic, but the mechanisms involved are not clear.     

The effect of MS 222 an anaesthetic on the peroxide metabolism enzymes in erythrocytes of freshwater and marine fish species

1. The effect of 70 mg/l and 35 mg/l MS 222 an anaesthetic on the enzymes: superoxide dismutase (SOD), catalase (C) and peroxidase (P) were estimated in erythrocytes of Cyprinus carpo, a freshwater fish and Dicentrarchus labrax, a marine fish. 2. The end of the summer, at 16 degrees C MS 222 in concentration 70 mg/l caused an enhancement of the SOD and peroxidase activities and a decrease of the catalase activity. 3. In the autumn at 22 degrees C SOD and peroxidase activities in erythrocytes of Dicentrarchus labrax are normally higher than at 16 degrees C. On the contrary MS 222 causes no significant modification of enzymatic activities measured, but an increase in the dispersion of the results. 4. At 13 degrees C in the spring, MS 222 has no immediate influence on the activity of these enzymes, whilst at the same temperature at the beginning of winter, SOD is the only one activated. 5. It seems that in experiments concerning peroxide metabolism enzymes the use of anaesthetic MS 222 is not advisable.     

Haematological evaluation of the anaesthetic benzocaine hydrochloride in the freshwater fish Cyprinus carpio L.

The effects of anaesthetization with different concentrations of benzocaine hydrochloride (BH) and neutralized benzocaine hydrochloride (NBH) were studied on the haematology of Cyprinus carpio. Due to its acidic nature and resultant effects on aquarium water, BH produces haemoconcentration effects with disturbances in acid-base function. The use of NBH, whereby water quality effects were drastically reduced, improved the general haematological profile. In contrast, haemodilution resulted when blood was sampled without the use of an anaesthetic agent.     

The uptake of the anaesthetic benzocaine hydrochloride by the gills and the skin of three freshwater fish species

The uptake of benzocaine hydrochloride and neutralized benzocaine hydrochloride by the skin and the gills of Cyprinus carpio, Oreochromis mossambicus and Salmo gairdneri were studied. The differences observed can mainly be ascribed to degree of ionization and the lipid solubility of the anaesthetic.     

Effects of anaesthesia with MS 222, neutralized MS 222 and benzocaine on the blood constituents of rainbow trout,Salmo gairdneri†

Rainbow trout (Salmo gairdneri Richardson) were subjected to 15 min anaesthesia with unbuffered MS 222, neutralized MS 222 and benzocaine with and without physical stress. Blood samples were taken through cannulae inserted into the dorsal aorta. The Hct values and Hb concentrations increased with all the anaesthetics, which also caused swelling of erythrocytes. The initial values were restored within 4–12 h of recovery. Each anaesthetic elevated the blood lactate concentration, but the initial level was regained within 12 h. The blood glucose level decreased the most during anaesthesia with unbuffered MS 222, but the initial level was rapidly restored. Benzocaine caused the least hypoglycaemia during anaesthesia, but the subsequent hyperglycaemia, as in the fish anaesthetized with neutralized MS 222, lasted 24 h. Neutralized MS 222 and benzocaine elevated the plasma K + concentration more rapidly than unbuffered MS 222. The initial levels were regained in 4 days. All anaesthetics raised the Mg ⁺⁺ concentration. The Po₂ in the dorsal aorta decreased during anaesthesia with unbuffered MS 222 by about 85 mmHg, while the Pco₂ increased by about 1.5 mmHg. Their initial levels were regained within 20 min. During anaesthesia the pH value decreased by 0.3 units and returned to the initial value within 2–4 h of recovery. MS 222 seemed to be an asphyxiant.     

The environmental preferences of three species of Australian freshwater fish in relation to the effects of riparian degradation

To assess the likely effects on three coexisting species of Australian freshwater fish of riparian loss, we examined the temperature, light, and habitat preferences of each species in relation to commonly documented effects of riparian degradation on stream environments. Such effects include reduced shade, instream structure, and water depth and increased temperature and invasive instream vegetation. Xiphophorus helleri, Gambusia holbrooki, and Melanotaenia duboulayi differed significantly in mean swimming depth, preferences for light and habitat, and in their patterns of behavioural change through the day. Values of interspecific spatial overlap (25–58%) indicated that the overall environmental preferences of G. holbrooki, X. helleri, and M. duboulayi were reasonably distinct. Habitat alterations associated with riparian removal are likely to favour the exotic species G. holbrooki over the native species M. duboulayi, but the results for X. helleri suggest that not all poeciliid␣species are strong indicators of degraded conditions.     

The physiological responses of blood during thermal adaptation in three freshwater fish species

Sarotherodon mossambicus , Cyprinus carpio and Salmo gairdneri were acclimatized at temperatures of 15, 20 and 25° C in order to study physiological responses of blood to temperature fluctuations in the laboratory. Cyprinus carpio exhibited the greater ability to survive at these temperatures. Sarotherodon mossambicus experienced osmoregulatory collapse at 15° C which also occurred in trout at 25° C. This was associated with acid-base malfunction in the trout.     

Hormonal and ion regulatory response in three freshwater fish species following waterborne copper exposure

We evaluated effects of sublethal copper exposure in 3 different freshwater fish: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gibel carp (Carassius auratus gibelio). In a first experiment we exposed these fishes to an equally toxic Cu dose, a Cu level 10 times lower than their 96 h LC₅₀ value: 20, 65, and 150 µg/L Cu. In a second series we exposed them to the same Cu concentration (50 µg/L). Na⁺/K⁺-ATPase activity in gill tissue was disturbed differently in rainbow trout then in common and gibel carp. Rainbow trout showed a thorough disruption of plasma ion levels at the beginning of both exposures, whereas common carp and gibel carp displayed effects only after 3 days. Rainbow trout and common carp thyroid hormones experienced adverse effects in the beginning of the exposure. The involvement of prolactin in handling metal stress was reflected in changes of mRNA prolactin receptor concentrations in gill tissue, with an up regulation of this mRNA in rainbow trout and a down regulation in gibel carp, which was more pronounced in the latter. Overall, rainbow trout appeared more sensitive in the beginning of the exposure, however, when it overcame this first challenge, it handled copper exposure in a better manner then common and gibel carp as they showed more long term impacts of Cu exposure.     

Comparative analysis of reproductive traits in 65 freshwater fish species: application to the domestication of new fish species

Based on an extensive literature search (1,000 references), the objectives of the present study were to establish a numerical clustering of temperate freshwater fish based on their reproductive traits and to evaluate whether it was possible to extrapolate zootechnical knowledge among species belonging to the same cluster. About 65 species were classified into ten homogeneous clusters from the analysis of 29 reproductive traits, among which the most important were temperature during spawning, egg incubation and larval rearing, degree-days for incubation, larval size upon hatching, spawning season, and parental care. From this typology, a rather regular continuum of reproductive clusters emerges with two obvious endpoints. Between these two extremes, species could be ordered chiefly according to temperature requirement, spawning season and parental care. In conclusion, this new typology, differing significantly from all others proposed earlier, may now serve as a possible framework to help enhancing the domestication of new species by comparison to species belonging to the same cluster.     

Fish-SPRICH: a database of freshwater fish species richness throughout the World

There is growing interest in large-scale approaches to ecology, for both plants and animals. In particular, macroecological studies enable examination of the patterns and determinants of species richness of a variety of groups of organism throughout the world, which might have important implications for prediction and mitigation of the consequences of global change. Here, we provide richness data for freshwater fishes, which, with more than 13,000 described species, comprise a quarter of all vertebrate species. We conducted an extensive literature survey of native, non-native (exotic), and endemic freshwater fish species richness. The resulting database, called Fish-SPRICH, contains data from more than 400 bibliographic sources including published papers, books, and grey literature sources. Fish-SPRICH contains richness values at the river basin grain for 1,054 river basins covering more than 80% of the earth’s continental surface. This database is currently the most comprehensive global database of native, non-native and endemic freshwater fish richness available at the river basin grain.     

The genetic structure of fish: differences in the intraspecific distribution of biochemical genetic variation between marine, anadromous, and freshwater species

Biochemical genetic markers have been increasingly used for inferences on the population genetic structure of various fish, but little attention has been paid to relative differences in the distribution of variability within species in different groups of fish. Nested gene diversity analyses of multiple locus electrophoretic data from the literature for 19 species (6 marine species, 4 anadromous salmonids, and 9 freshwater species (5 salmonids)), mainly from Scandinavia and North America, show that (i) average heterozygosity is higher in marine species than in freshwater species, (ii) the average fraction of the total gene diversity allocated between localities increases in the order marine (1.6%), anadromous (3.7%), and freshwater species (29.4%), and (iii) the fraction of the total gene diversity associated with differences between geographic clusters of localities is generally smaller than that between localities within clusters. This finding is consistent with that the differentiation observed in most species is of postglacial origin, and that the timespan available since deglaciation has been too short to result in the accumulation of substantial differences between clusters.     

Toxicity of intermittent chlorination to freshwater fish: Influence of temperature and chlorine form

Fingerling size Salmo gairdneri, Oncorhynchus kisutch, Notemigonus crysoleucas, Cyprinus carpio, and Ictalurus punctatus were exposed in the laboratory three times daily for up to seven days to pulses of either free chlorine or monochloramine. This regime simulated conditions often encountered in the outfall of steam electric generating plants which chlorinate intermittently. LC₅₀'s, LT₅₀'s and response isopleths giving various percentage mortalities, were computed from the bioassays. S. gairdneri, O. kisutch, and I. punctatus were the most sensitive to both types of chlorine. C. carpio were most resistant and the N. crysoleucas were intermediate in sensitivity. Temperature had relatively little effect on the toxicity of intermittent chlorine to the species tested. In this type of test regime, free chlorine was three to fourteen-fold more toxic (depending on the species) than monochloramine. Water quality criteria for the protection of fish should, in the future, take this differential toxicity into consideration.     

Midazolam and fentanyl-fluanisone: assessment of anaesthetic effects in laboratory rodents and rabbits

The anaesthetic effects of a combination of midazolam, a water-soluble benzodiazepine, and fentanyl-fluanisone were assessed in laboratory rodents and rabbits. This combination of drugs produced good surgical anaesthesia in all of the species investigated. An important advantage of the combination was that it could be administered as a single intraperitoneal injection.     

Ontogenetic changes in the toxicity and efficacy of the anaesthetic MS222 (tricaine methanesulfonate) in zebrafish (Danio rerio) larvae

Median lethal (LC(50)) and effective (EC(50)) concentrations for 1-h and 24-h exposures to the anaesthetic MS222 (tricaine methanesulfonate) were determined for zebrafish Danio rerio larvae ranging in age from 3 days postfertilization (dpf) to 9 dpf. Cessation of heart beat was used as the indicator of death (LC(50)) while failure to respond to direct mechanical stimulation of the head region was taken as an indication of deep anaesthesia (EC(50)). 1-h LC(50)s, 1-h EC(50)s and 24-h EC(50)s all decreased gradually but significantly (all P<0.01) with age. Mean values for 1-h LC(50)s were 1633 mg L(-1) and 730 mg L(-1), respectively, for 3 dpf and 9 dpf larvae. Mean value for 1-h and 24-h EC(50)s were 106 mg L(-1) and 100 mg L(-1), respectively, at 3 dpf and 65 mg L(-1) and 31 mg L(-1), respectively, at 9 dpf. The gradual increase with age in sensitivity to the anaesthetic implied by these indicators is probably a reflection of ontogenetic changes in the activity of detoxification pathways. Mean values for the 24-h LC(50) also decreased significantly (P<0.001) with age, from 566 mg L(-1) at 3 dpf to 64 mg L(-1) at 9 dpf. However, unlike the other indicators, the decrease was not gradual but occurred in a step-like fashion with virtually all of the change occurring between 4 dpf and 7 dpf. This sharp increase in sensitivity coincides with the shift in the major site of systemic ionoregulatory activity from the skin to the gills. The implications of these ontogenetic changes in lethal and effective levels for researchers or others intending to use the anaesthetic with fish larvae are discussed.     

Conservation of freshwater fish resources of India: new approaches, assessment and challenges

The freshwater resources of India are currently experiencing an alarming decline in fish biodiversity due to several factors and as a result, a sizeable portion of fresh water fishes have been categorized as threatened. This emphasizes an immediate need for initiating research and actions for alternative management techniques to protect these aquatic systems. One such option that has potential to protect freshwater ecosystem from numerous threats is the creation of freshwater aquatic sanctuary (FAS) within protected area network. Though similar conservation practices are well established in the terrestrial and marine ecosystem, however, the work on freshwater systems has been very slow and negligible. In the present communication we conceptualized the need and approach for developing FAS within the protected area network based on our observations in the water bodies of the selected wildlife sanctuaries in Northern India as well as success stories of some other countries. In this study we assessed the fish diversity in the selected protected areas of Northern India. The assessment indicated that these sanctuaries harbor 28.26–31.13% of freshwater fishes, which are threatened in other areas. Apart from Indian Major Carps, Tor putitora, Chitala chitala, Pangasius pangasius, Clupisoma gerua, Ailia coila, Aorichthys aor, Wallago attu, Rhinomugil corsula, Ompok pabda, Ombok pabo etc. were the important species encountered in the protected waters. The various issues related to FAS including objectives, approach, potential tools, implementation and management are discussed towards saving endangered fish germplasm resources. Approaches, tools and modus operandi proposed in this communication could be utilized by other developing countries in the region.     

Effective reassessments of freshwater fish species: a case study in a Mediterranean peninsula

Hydrobiologia - IUCN Red List assessments for fish species can quickly become out of date. In recent years molecular techniques have added new ways of obtaining information about species...     

Morphological variation of the digestive tract: a feeding behaviour response in a freshwater fish species

Environmental Biology of Fishes - Plasticity of resource use represents an important strategy for fish species living in unstable freshwater environments. Trophic polymorphism (i.e. variation in...