A morphological and cytochemical study of erythrocytes, thrombocytes and leukocytes in four freshwater teleosts

Erythrocytes, thrombocytes and leukocytes morphology and cytochemical staining were studied in big head carp Aristichthys nobilis , oscar Astronotus ocellatus , traíra Hoplias malabaricus and lambari Astyanax bimaculatus . Reticulocytes contained a granular material similar to residual RNA following staining with brilliant cresyl blue. Neutrophils, lymphocytes and monocytes were morphologically similar in all the four species. Thrombocytes were present in all the four species and were predominantly fusiform, whereas eosinophils occurred only in A. ocellatus . Aristichthys nobilis contained a leukocyte with unstained granules following Romanowsky-type staining, which stained intensely with periodic acid-Schiff (PAS). Glycogen granules were present in thrombocytes, neutrophils and eosinophils but not in monocytes or lymphocytes. Peroxidase staining was observed in neutrophils of A. ocellatus , H. malabaricus and A. bimaculatus but not in A. nobilis . Monocytes of A. ocellatus , H. malabaricus and A. bimaculatus stained positively for non-specific esterase, whereas those of A. nobilis did not stain. Thrombocytes and leukocytes in all four species were negative for alkaline phosphatase. Neutrophils of A. ocellatus and H. malabaricus may be involved in respiratory burst and may play an important microbicidal role.     

Nitrogen and phosphorus content of some temperate and tropical freshwater fishes

The nitrogen and phosphorus content of two temperate fishes, Rutilus rutilus and Perca fluviatilis , and six tropical fishes, Oreochromis niloticus , Cichla monoculus , Serrassalmus rhombeus , Plagioscion squamosissimus , Prochilodus brevis and Hoplias malabaricus , were investigated to test the hypothesis that variation in body P content and N:P ratio is related to body size. Regressions of %P and N:P ratios against fish size (length and mass) confirmed the hypothesis for P. fluviatilis and P. squamosissimus , suggesting that body size is an important factor driving body P content and N:P ratios in some fishes. Moreover, significant increases in %N and N:P ratio with body size was found for H. malabaricus , a common piscivorous fish of the Neotropics. Interspecific variation in %P and N:P ranged two-fold and significant differences ( P < 0·05) were found among the tested species. The mean ± s . d . elemental content across all fishes ( n = 170) was 10·35 ± 1·29% for N and 3·05 ± 0·82% for P, while the N:P ratio was 8·00 ± 2·14. Data on fish body nutrient content and ratio will improve parameterization of bioenergetics and mass balance models and help clarify the role of fishes in nutrient cycles in both temperate and tropical freshwaters.     

The fish fauna in tropical rivers: the case of the Sorocaba River basin, São Paulo, Brazil

A survey was carried out on the fish species in the Sorocaba River basin, the main tributary of the left margin of the Tietê River, located in the State of S?o Paulo, Brazil. The species were collected with gill nets. After identification of the specimens, their relative abundance, weight and standard length were determined. Up to the present moment there are not any studies that focus this subject in this hydrographic basin. Fifty-three species, distributed in eighteen families and six orders were collected. Characiformes were represented by twenty-eight species, Siluriformes by seventeen species, the Gymnotiformes by three species, Perciformes and Cyprinodontiformes by two species, and the Synbranchiformes by one species. Among the collected species there were two exotic. The most abundant species were Astyanax fasciatus and Hypostomus ancistroides. In relation to total weight the most representative species were Hoplias malabaricus and Hypostomus ancistroides. Cyprinus carpio, Prochilodus lineatus, Schizodon nasutus and Hoplias malabaricus were the most representative species in relation to average weight. Largest standard length were recorded for Sternopygus macrurus, Steindachnerina insculpta, Eigenmannia aff. virescens and Cyprinus carpio.     

The protective influence of ascorbic acid against the genotoxicity of waterborne lead exposure in Nile tilapia Oreochromis niloticus (L.)

The present study investigated the effects of lead (Pb) and ascorbic acid (AA) on Nile tilapia Oreochromis niloticus using the micronucleus (MN) and nuclear abnormality (NA) tests for periods of 7, 14 and 21 days. The MN frequencies in the erythrocytes, gill, liver and fin cells were analysed comparatively to evaluate the sensitivity and suitability of these different cell types. The NA shapes in erythrocytes were scored into blebbed nuclei (BL), lobed nuclei (LB), notched nuclei (NT) and binuclei (BN). It was observed that fish showed significant sensitivity to the different treatments. In general, the highest value of both MN and NA cells were significantly increased in the Pb-treated group followed by the combination of the Pb and AA-treated group. On the other hand, the MN and NA frequencies in erythrocytes were the most sensitive to the treatment and could provide more valuable information than those in gill, liver and fin cells. The frequencies of each NA shape in erythrocytes of all treatments were observed in the following ranked order NT > LB > BN > BL. The results demonstrated the efficacy of AA in reducing genotoxicity in fish induced by Pb. They showed the sensitivity and suitability of MN and NA frequencies in erythrocytes as pollution biomarkers.     

Karyological evidence for interspecific hybridization between Cichla monoculus and C. temensis (Perciformes, Cichlidae) in the Amazon

Cichla monoculus, Cichla temensis (peacock bass or tucunare), and its presumed hybrids, were cytogenetically analyzed. The fish were collected at three distinct sites in the central Amazon basin, namely in the Uatuma (C. monoculus, C. temensis and their natural hybrid), Jau (C. temensis), and Solimoes rivers (C. monoculus). The two species and the natural hybrid showed the same diploid number, 2n=48 acrocentric chromosomes. Single NORs were detected in the distal region of the long arm in all three species. However, in C. monoculus, the NOR was found on the second pair of the complement, in C. temensis, on the third pair and in the hybrid two NOR patterns were found, one on the second pair and the other on the third pair of chromosomes. The two species and the hybrid have their constitutive heterochromatin located in the pericentromeric region of all chromosomes and an interstitial C-band located on the largest chromosome pair. The great similarity in the chromosome number and morphology, chromosome size class differences, the NOR patterns and C-banding suggested chromosomal stasis during speciation and hybridization of Cichla.     

Spatial synchrony of a highly endemic fish assemblage (Segredo Reservoir, Igua?u River, Paraná State, Brazil).

In this study, patterns of spatial synchrony in population fluctuations (cross-correlation) of an endemic fish assemblage of a Neotropical reservoir (Segredo Reservoir, Igua?u River, Paraná State, Brazil) were reported. First, the level of population synchrony for 20 species was estimated. Second, population synchrony was correlated, using the Mantel test, with geographical distances among sites (n = 11) and also environmental synchrony (temperature). Nine species presented significant correlations between spatial synchrony and geographic distances (Astyanax sp. b, Astyanax sp. c, Pimelodus sp., Hoplias malabaricus, Crenicichla iguassuensis, Hypostomus derbyi, Hypostomus myersi, Rhamdia branneri, and R. voulezi). Considering the ecology of the species and the significant relationship between population and environmental synchronies, it seems that environmental stochasticity is the most plausible hypothesis in explaining the observed synchrony patterns.     

Evidence for a natural hybrid of peacock bass (Cichla monoculus vs Cichla temensis) based on esterase electrophoretic patterns

Esterase (Est) and esterase-D (Est-D) electrophoretic patterns identified by starch gel electrophoresis of skeletal muscle protein extracts of 184 specimens of three species of peacock bass, locally known as tucunares (Cichla monoculus, C. temensis and Cichla sp), plus four specimens of a supposed hybrid (C. monoculus vs C. temensis), collected from the Central Amazon, were examined to determine if they could aid in identifying a supposed hybrid between C. monoculus and C. temensis. Six zones of electrophoretic activity were found with these enzyme systems. The Est enzyme showed one zone of activity, formed by bands 1, 2 and 3, plus three zones of activity, presumably controlled by Est-1, 2 and 3 loci. The Est-D enzyme had two zones of activity, presumably controlled by Est-D1 and Est-D2 loci. Cichla monoculus and C. temensis shared band 2 and alleles Est-1(1), Est-2(1), Est-3(2), and Est-D1(1), and therefore these were useless for identifying hybrids between the two species. However, a probable hybrid pattern of bands 1, 2, and 3, presumably generated by a combination of pattern 12 from C. monoculus with pattern 23 from C. temensis, resulting from a possible cross between these two species, was detected. Although the Est-D2 locus cannot be considered an ideal diagnostic marker for identifying the supposed hybrid (C. monoculus vs C. temensis), as it is polymorphic, it proved to be useful for determining the origin of the hybrid, i.e., which parental species were involved in the hybridization process.     

Respiratory responses to hypoxia in relation to mode of life of two erythrinid species (Hoplias malabaricus and Hoplias lacerdae)

Predominantly, Hoplias malabaricus inhabits stagnant O₂ poor environments, whereas Hoplias lacerdae occurs in well-aerated streams. The present study evaluates the influence of mode of life on O₂ uptake and gill ventilation in equally-sized (300 g) specimens of this genus at 25° C. Comparing the species, H. lacerdae was characterized by the highest O₂ uptake and gill ventilation combined with a relatively higher cost of breathing and a lower O₂ extraction. Both species substantially increased ventilation in response to hypoxia with the difference that H. malabaricus exclusively augmented tidal volume, whereas H. lacerdae also increased breathing frequency.     

In situ evaluation of the genotoxic potential of the river Nile: II. Detection of DNA strand-breakage and apoptosis in Oreochromis niloticus niloticus (Linnaeus, 1758) and Clarias gariepinus (Burchell, 1822)

This work is part of a wider eco-toxicological study proposed to evaluate the biological impact of contaminants along the whole course of the river Nile, Egypt. Here we present data on the presence of DNA strand-breaks and apoptotic cells assessed by use of comet and diffusion assays in erythrocytes of Nile tilapia (Oreochromis niloticus niloticus) and African catfish (Clarias gariepinus). The results showed high degrees of DNA damage and increased frequencies of apoptotic nuclei in blood of fish collected from downstream compared with those sampled from upstream river Nile. Qualitative analysis revealed a shift in the frequency of DNA-damage classes towards higher damage levels correlating with the increasing pollution gradient. The degree of DNA damage measured by use of comet assay and diffusion assay exhibited seasonal variations. Both fish species showed significant increases in DNA damage during the summer. The results of our study indicated that the alkaline comet assay seems to be a useful technique for in situ genotoxic monitoring. At the same time the diffusion assay is sensitive enough to detect low frequencies of apoptotic nuclei. The results reveal species-specific differences in sensitivities, suggesting that Nile tilapia may serve as a more sensitive test species compared with the African catfish. Based on the outcome of the comet and diffusion assays, it can be concluded that the water quality of the river Nile with respect to the presence of genotoxic compounds needs to be improved, especially in its estuaries. As far as we know this is the first time that the comet and diffusion assays are used for genotoxic monitoring of the river Nile.     

Temporal patterns of resource partitioning among Cichla species in a Venezuelan blackwater river

In channel and floodplain habitats of the Cinaruco River, Venezuela, Cichla temensis was more abundant and larger than C. intermedia and C. orinocensis . Seasonal variation in hydrology influenced habitat use, spawning, and predator-prey interactions. The three piscivores partitioned habitat, with C. intermedia showing a strong affinity for structured habitats in the main channel during all water level fluctuations. C. orinocensis was most abundant in shallow areas with submerged structure in lagoons and, to a lesser extent, in low velocity regions of the channel, and C. temensis occupied a wide range of lotic and lentic habitats. During the low-water period, the feeding frequency and body condition of all three species declined, and this was related, in part, to preparation for spawning near the end of the low-water season. The diet of C. intermedia was least similar to its two congeners during falling and rising water. C. orinocensis and C. temensis had lowest diet overlap during the low-water conditions, the period when many individuals of these two species move into lagoons for nesting. Prey in stomachs were significantly larger during the falling-water than the rising-water period, and predation by Cichla and other large piscivores during the falling-water period may have reduced the abundance of large prey, particularly Semaprochilodus kneri . These migratory detritivorous fish were important prey for C. temensis during the falling-water period and probably contributed a substantial fraction of the annual energy intake for this species. Together, the three Cichla species consume a wide spectrum of prey from a diverse fish assemblage, but prey are subdivided based on habitat, prey type, and season.     

Micronuclei monitoring of fishes from Lake Paranoá, under influence of sewage treatment plant discharges

Fish micronuclei tests (MN) were used to evaluate the ability of wastewater from two municipal sewage treatment plants that empty into Lake Paranoá to cause genetic damage. There were no significant differences in the frequencies of micronuclei between the control and hypertrophic areas. In contrast, cyclophosphamide and mitomycin C, used to test the sensitivity of the biological assay, significantly increased the micronuclei counts in Tilapia rendalli, Oreochromis niloticus and Cyprinus carpio, T. rendalli was the most sensitive specie to both clastogens and C. carpio, the most resistant.     

Ureotelism is inducible in the neotropical freshwater Hoplias malabaricus (Teleostei, Erythrinidae).

Increased environmental pH decreases ammonia transport through the gills, impairing nitrogenous waste. The consequent toxicity is usually drastic to most fishes. A few species are able to synthesize urea as a way to detoxify plasma ammonia. We studied three teleosts of the family Erythrinidae living in distinct environments, and assumed the biochemical behaviors would be different in spite of their being closely related species. Adult fish collected in the wild were submitted to alkaline water and the urea excretion rate was determined. The specific activity of urea cycle enzymes was determined in liver samples of fish from neutral waters. The studied species Hoplias lacerdae, Hoplerithrynus unitaeniatus, and Hoplias malabaricus are ureogenic. Urea synthesis is not a metabolic way to detoxify ammonia in H. lacerdae and Hoplerithrynus unitaeniatus exposed to an alkaline environment. The plasma ammonia profile of both species showed two distinct biochemical responses. Urea excretion of H. malabaricus was high in alkaline water, and the transition to ureotelism is proposed. The nitrogen excretion rate of H. malabaricus was among the highest values reported and the high urea excretion leads us to include this species as ureotelic in alkaline water.     

The tilapiine fish stock of Lake Victoria before and after the Nile perch upsurge

Since the beginning of fisheries in Lake Victoria, two native tilapiine species, Oreochromis esculentus and Oreochromis variabilis , were the main target of the local fishermen. A continuous increase in fishing pressure led initially to a declining catch per unit of effort, and a smaller average fish size; eventually, there was a reduced landing of tilapiines. To boost the fisheries, three alien tilapiine species and the Nile perch Lates niloticus were introduced. Thirty years after its introduction, Oreochromis niloticus appeared to be the most successful tilapiine species. It replaced the indigenous tilapiines almost completely before the Nile perch came to dominate the ecosystem of Lake Victoria. Reduced fishing pressure on the tilapiines in the 1980s, due to the shift of the local fishery towards the Nile perch, resulted in an increase in the stock of O. niloticus and an increase in average fish size. Subsequently, the total mass of O. niloticus landed increased. The stocks of the indigenous tilapiines did not recover but declined to extremely low levels, or vanished from the main lake. Currently, these species still occur in satellite lakes of Lake Victoria, from which O. niloticus is absent. Nile perch feed on O. niloticus; however, the limited overlap in distribution between piscivorous Nile perch and O. niloticus of consumable sizes is probably an important factor in explaining the coexistence of the two species. The main cause of the disappearance of the native tilapiine species is presumed to be competitive dominance by O. niloticus .     

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 April 2012 - 31 May 2012

This article documents the addition of 123 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Brenthis ino, Cichla orinocensis, Cichla temensis, Epinephelus striatus, Gobio gobio, Liocarcinus depurator, Macrolophus pygmaeus, Monilinia vaccinii-corymbosi, Pelochelys cantorii, Philotrypesis josephi, Romanogobio vladykovi, Takydromus luyeanus and Takydromus viridipunctatus. These loci were cross-tested on the following species: Cichla intermedia, Cichla ocellaris, Cichla pinima, Epinephelus acanthistius, Gobio carpathicus, Gobio obtusirostris, Gobio sp. 1, Gobio volgensis, Macrolophus costalis, Macrolophus melanotoma, Macrolophus pygmaeus, Romanogobio albipinnatus, Romanogobio banaticus, Romanogobio belingi, Romanogobio kesslerii, Romanogobio parvus, Romanogobio pentatrichus, Romanogobio uranoscopus, Takydromus formosanus, Takydromus hsuehshanesis and Takydromus stejnegeri.     

Systematics, biogeography, and evolution of the Neotropical peacock basses Cichla (Perciformes: Cichlidae)

To investigate forces influencing diversification in Neotropical fishes, the phylogenetic relationships among species and populations of the cichlid genus Cichla were examined. Mitochondrial DNA was sequenced for 454 individuals of the 5 nominal Cichla species and several putative undescribed species. Phylogenetic analyses support the distinction of two major clades of Cichla. Clade A includes C. temensis and two undescribed species from the lower Amazonas and Xingu Rivers. Clade B includes C. orinocensis, C. monoculus, C. ocellaris. C, intermedia, and an undescribed species from the upper Madeira River. Species boundaries were relatively well-circumscribed for clade B, while incomplete lineage sorting was inferred for clade A. Three probable instances of introgression were observed, including a regional population of C. orinocensis from the Negro River that shows a history of introgression. Biogeographic patterns from Cichla are partially congruent with those seen in several other Neotropical fish clades, and the diversification of Cichla species is inferred to result from both vicariance and sympatric divergence.     

Effects of temperature on prey consumption and growth in mass of juvenile trahira Hoplias aff. malabaricus (Bloch, 1794)

The influence of temperature on prey consumption and growth in mass of juvenile trahira Hoplias aff. malabaricus were investigated. Consumption of small-sized lambari Astyanax altiparanae (mean standard length, L _S, 5·43 cm) varied from zero to 65 over a period of 30 days. Temperatures ranged from 14 to 34° C and the size of trahiras ranged from 17·5 to 24·7 cm L _S. Prey consumption differed significantly among temperatures. Trahiras at 18° C consumed significantly less than those at 30° C. A linear multiple regression model including temperature, prey consumption and L _S explained 89·4% of the variability in growth in mass. Some caution is suggested when inferring the impact of H. aff. malabaricus piscivory on assemblage structures in systems that, despite their location in tropical regions, are subjected to seasonal thermal variations.     

Influence of life history and seasonal hydrology on lipid storage in three neotropical fish species

The seasonal dynamics of energy (lipid) storage in three neotropical fish species with differing life histories were evaluated. Lipid content was substantially greater in the liver than in dorsal musculature of all three species. Two piscivores ( Cichla temensis and Serrasalmus manueli ) showed large, statistically significant seasonal fluctuations in liver lipid content. Liver lipid content increased during high water, through the falling water, and into the early dry season for both piscivores. Seasonal variation in dorsal muscle lipid content was large and statistically significant for C. temensis , but was small and non‐significant for S. manueli . Cichla temensis appeared to 'finance' costs associated with reproduction by accumulating lipids during the falling‐water period when migratory prey allowed the species to subsidize their energetic dynamics. Semaprochilodus kneri , a migratory algivore and detritivore, showed no significant seasonal variation in dorsal muscle lipid content and minimally significant seasonal variation in liver lipid content. Statistically significant effects of lipid content on δ¹³C was observed when tissue lipid content varied by >12%, while biological interpretation of food web statistics based on δ¹³C values appears robust to minor variations in lipid content. Nonetheless, when lipid content varied by larger amounts ( e.g . >35% for C. temensis and S. manueli liver tissue) lipids appeared to have a large potential effect on δ¹³C and food web statistics calculated from such measurements may have been biased. Surprisingly, even large variation in tissue lipid content did not affect δ¹⁵N.     

Toxicity and genotoxicity of wastewater from gasoline stations

The toxicity and genotoxicity of wastewater from eight gasoline stations in Brasília, Brazil's capital city, was studied by assessing chromosomal aberrations, chromosomal malsegregation and the mitotic index in Alliumcepa root cells, and the occurrence of micronucleus and nuclear abnormalities in peripheral erythrocytes of tilapia fish (Oreochromis niloticus). The content of gasoline station effluents was also analyzed based on several physico-chemical parameters. None of the wastewater samples was genotoxic to A. cepa root cells, although cell proliferation was significantly inhibited, especially at the highest concentrations. Likewise, no micronuclei were observed in O. niloticus peripheral erythrocytes, even after exposure to high concentrations, but there was an increase in the number of nuclear abnormalities and fish mortality. These results show that although the effluent from gasoline stations is processed by an oil/water separation system before being discharged into the main sewage system, the wastewater still contains toxic compounds.     

Recent introgressive hybridization revealed by exclusive mtDNA transfer from Oreochromis leucostictus (Trewavas, 1933) to Oreochromis niloticus (Linnaeus, 1758) in Lake Baringo, Kenya

Nuclear DNA and mtDNA polymorphisms were surveyed in various species of East African Oreochromis . In Lake Baringo, where only Oreochromis niloticus baringoensis is present, alien mtDNA haplotypes were observed, apparently the result of introgressive hybridization with Oreochromis leucostictus . This introgression is not accompanied by any substantial or recorded transfer of nuclear genes into O. n. baringoensis .     

Age structure and growth of peacock cichlids from rivers and reservoirs of Venezuela

Cichla temensis collected in Venezuela attained a mean standard length of 440 mm, and C. orinocensis and C. intermedia mean lengths of 309 and 321 mm, respectively. There was little evidence of size differences between sexes. Opaque zones on otoliths corresponded to the period of peak of reproductive activity and low fish body condition ( K ). When aggregated across sites, linear estimates for growth of fish >age I were 57·5, 27·6, and 19·6 mm year¹ for C. temensis , C. orinocensis , and C. intermedia , respectively. Fish from the Cinaruco River appeared to spawn at age I, and prior to first maturation had grown 25, 21, and 23 mm month¹, respectively, for C. temensis , C. orinocensis , and C. intermedia . There was little intersite variation in growth rates, and differences in sizes of conspecifics from different sizes were due to age differences. Cichla in captivity spawn several times per year. The present results indicate that the annual cycle or tropical wet/dry seasonality promotes reproductive seasonality in Cichla . The energetic cost of reproduction and brood defence probably constrains most individuals to one spawn per year. Conversely, populations living in relatively aseasonal reservoir environments may have several spawns per year, in which case growth zones on hard structures are difficult to interpret.