Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for ∼10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare. © 2002 Elsevier Science B.V. All rights reserved.

Effect of fish density during transportation on stress and mortality of juvenile Tambaqui Colossoma macropomum

The increased demand for juvenile tambaqui Colossoma macropomum for grow-out ponds and stocking programs in the Amazon state of Brazil has increased the transportation of this species. This study was designed to determine the optimum density of juvenile tambaqui during transportation in closed containers. Fish (51.9 ± 3.3 g and 14.9 ± 0.4 cm) were packed in sealed plastic bags and transported for 10 h at four densities: 78, 156, 234, and 312 kg/m3. After transportation, fish from each density were kept in separate 500-L tanks for 96 h. Mortality, 96-h cumulative mortality, water quality, and blood parameters (hematocrit, plasma cortisol, and glucose) were monitored. Fish mortality after transportation was significantly lower at densities of 78 and 156 kg/m3 than at 234 and 312 kg/m3. Cumulative mortality was significantly lower at a density of 78 kg/m3. Dissolved oxygen after 10 h of transportation remained high at a density of 78 kg/m3, but reached critically low values at all other densities. Ammonia concentration was highest at the lowest density and was lower at higher densities. Carbon dioxide concentration was lowest at the density of 78 kg/m3 but higher in the other treatments. Plasma glucose and cortisol increased significantly immediately after transportation at densities of 156, 234, and 312 kg/m3, returning to control values by 24 h. The best density for juvenile tambaqui during a 10-h transportation haul in a closed container was 78 kg/m3. At this density there was no fish mortality, water quality was kept within acceptable values, and fish were not stressed.

Short interspersed repetitive elements (SINEs) from the cichlid fish, Oreochromis niloticus, and their chromosomal localization by fluorescent in situ hybridization

In the present study, we describe the cloning and characterization of a new SINE-like element from O. niloticus (ROn-2) and show the distribution of this SINE and a previously isolated SINE, ROn-1, in the chromosomes of O. niloticus. The ROn-2 element is 359 base pairs (bp) in length, contains short direct terminal repeats, a tRNA-related region similar to tRNA Val and tRNA Arg, a tRNA-unrelated region, and a poly-A tail. Analysis of the chromosomal distribution of ROn-1 and ROn-2 by fluorescent in situ hybridization showed that both SINE sequences are present in all chromosomes of tilapia, and organized in small clusters. The only exception was a large cluster of ROn-1 repeats found in the middle of the long arm of chromosome 1. In view of our data we discuss the hypothesis that the absence of large clusters of SINE sequences and the structural composition of these sequences may explain the absence of base-specific fluorochrome bands in the chromosomes of tilapia.

Factors affecting fish diversity and abundance in drying ponds and lagoons in the upper Paraná River basin, Brazil

Monthly samples were taken from April 1992 to March 1993 of fish assemblages present in six permanent lagoons and four ponds, fragmented from them during the drought period and during the terminal phases of desiccation, to evaluate changes in their diversity, abundance and dominance. Their relationships with hydrological and environment conditions were examined. A total of 63 fish species were identified. Species composition and abiotic factors were heterogeneous among the different water bodies. The proportion of piscivores and oxygen concentration were the determining factors for structuring the assemblages. The impact of piscivory upon fish diversity was neutral at the beginning and negative at the end of the dry season. In the terminal phase of desiccation, diversity was higher in ponds that contained a moderate proportion of piscivores than in these containing a higher proportion.

Animais aquáticos de importância médica no Brasil

The injuries caused by venomous and poisonous aquatic animals may provoke important morbidity in the victim. The cnidarians (jellyfishes, especially cubomedusas and Portuguese-Man-of-War) caused nearly 25% of 236 accidents by marine animals, while sea urchins were responsible for about 50% and catfish, stingrays and scorpionfish nearly 25%). In freshwater, stingrays and catfish cause injuries with a very similar mechanism to the poisoning and the effects of the toxins of marine species. In a series of about 200 injuries observed among freshwater fishermen, nearly 40% were caused by freshwater catfish, 5% freshwater stingrays and 55% by traumatogenic fish, such as piranhas and trairas. The author presents the aquatic animals that cause injuries to humans in Brazil, the clinical aspects of the envenoming and the first measures for the control of the severe pain observed mainly in the accidents caused by cnidarians and venomous fishes.

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 São 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 off. virescens and Cyprinus carpio.

Utilization of fish protein hydrolysate in prepared diets for pacu, Piaractus mesopotamicus, larvae

Growth and survival rates of pacu, Piaractus mesopotamicus, larvae fed prepared diets containing different animal protein sources were evaluated. Four diets with the same level of crude protein (CP) (36%) and calories (4.02 kcal gross energy/g of diet) were fed to the larvae. Diets were formulated to contain one of four protein sources: (1) fish meal (FM), (2) tilapia residue silage (TS), (3) protein hydrolysate from tilapia residue (HT), and (4) eviscerated tilapia residue (HET). Larvae were fed Artemia nauplii for six days, prior to the start of the study, and the prepared diet was supplied from day 7 until the study concluded. Variance analysis showed no significant differences (P > 0.05) for survival rates and larval final lengths among treatments. However, final average weights were significantly different (P < 0.05 for larvae fed FM and HT. Average survival rates were relatively high and ranged from 68.1% to 73.9%. After the live food was replaced by prepared diets, no larval growth was observed for any treatment. Fish protein hydrolysate (HT and HET) and fish silage showed potential to be used as ingredients in the diet of pacu larvae. However, hydrolysate inclusion levels, processing methods to minimize nutrient lixiviation, and the best moment to replace live food with an inert diet (weaning) need further investigation. © 2003 by The Haworth Press, Inc. All rights reserved.

Transforming open mining pits into fish farms: Moving towards sustainability

The legacy of mining activities has typically been land 'returned to wildlife', or, at some sites, degraded to such an extent that it is unsuitable for any alternate use. Progress towards sustainability is made when value is added in terms of the ecological, social and economic well-being of the community. In keeping with the principles of sustainable development, the innovative use of flooded open pits and tailings impoundments as commercial, recreational or ornamental fish farms should be considered in some locations, as it could make a significant contribution to the social equity, economic vitality and environmental integrity of mining communities. This article highlights the growing significance of aquaculture and explores the benefits and barriers to transforming flooded pits and impoundments into aquaculture operations. Among other benefits, aquaculture may provide a much-needed source of revenue, employment and, in some cases, food to communities impacted by mine closure. Further, aquaculture in a controlled closed environment may be more acceptable to critics of fish farming who are concerned about fish escapes and viral transmissions to wild populations. Despite the potential benefits, aquaculture in flooded pits and impoundments is not without its complications - it requires a site-specific design approach that must consider issues ranging from metals uptake by fish, to the long-term viability of the aquatic system as fish habitat, to the overall contribution of aquaculture to sustainability. © 2004 United Nations. Published by Blackwell Publishing.

Lipoic acid and ascorbic acid affect plasma free amino acids selectively in the teleost fish pacu (Piaractus mesopotamicus)

Most studies on the antioxidants, lipoic acid (LA) and ascorbic acid (AA), focused on species that, unlike teleost fish, are not scurvy-prone, and are able to synthesize AA. The antioxidant properties of LA may make it useful in aquaculture nutrition, but several effects must first be investigated, and we address here plasma free amino acids (FAA). In mammals, LA and AA in high doses were claimed to alter plasma FAA profile; to our knowledge, however, no data are available in fish. We therefore studied the effects of dietary LA and AA on plasma FAA in the South American teleost fish pacu, which is being used increasingly in aquaculture. LA treatment decreased concentrations of 18 of 23 individual FAA; specifically, dispensable and total FAA were significantly affected. Ornithine was elevated (+26%) in LA-treated fish and significantly decreased ratios of plasma [Arg]/[Orn] and other individual [FAA]/[Orn] were observed. LA and AA both affected sulfur FAA concentrations. Plasma cystine levels were significantly increased in the LA-supplemented groups. AA had little effect on most amino acids, and no interaction with LA was detected. AA supplementation did, however, significantly lower taurine (-42%) and cystathionine (-31%) levels in plasma. No effect on the branched chain:aromatic amino acid ratios was observed. The data indicate that at the dietary level studied, LA and AA independently affect selected plasma FAA in pacu, and suggest that any use of LA in particular as a dietary supplement should take into account an altered plasma FAA profile.

Feeding and condition factor of characidiin fish in Ribeirão Grande system, Southeastern Brazil

Feeding, seasonal changes in visceral fat and condition factor were compared in two species of characidiin fishes, Characidium lauroi and C. alipioi from Ribeirão Grande system, southeastern Brazil. Five streams of Ribeirão Grande system were sampled (22° 47' 08 S, 45° 28' 17W). The samples were taken four times per site, from July, 2001 to April, 2002: winter (July 2001), spring (October 2001), summer (February 2002) and autumn (April 2002). Quantitative collections were made with an electro-fishing device powered by a generator with maximum capacity of 1,500 V and 8.7 A of 60 Hz alternating current. Ephemeroptera nymphs, Diptera larvae (Chironomidae, Simuliidae), Plecoptera nymphs, Trichoptera larvae (Hydroptilidae, Psychoyiidae), terrestrial insects (Coleoptera, Isoptera, Hemiptera [Heteroptera, Homoptera]), Megaloptera larvae (Corydalidae), Arachnida, Ostracoda and vegetal debris were found in both species' diets. Visceral fat declined in February, coinciding with the decline of the condition factor in both species. The increased feeding from summer to fall provides fat accumulation. During subsequent seasons, fish may utilize visceral fat reserves for maintenance and reproduction.

Effect of dietary supplementation with vitamin E and stocking density on macrophage recruitment and giant cell formation in the teleost fish, Piaractus mesopotamicus

The effect of dietary supplementation with 0, 100 and 450 mg of vitamin E (DL-α tocopheryl acetate)/kg of a dry diet on the kinetics of macrophage recruitment and giant cell formation in the pacu, maintained at different stocking densities (5 kg/m3 and 20 kg/m3), was investigated by insertion of round glass coverslips into the subcutaneous connective tissue. After a feeding period of 18 weeks, the coverslips were implanted and later removed for examination at 2, 7 and 15 days post-implantation. Fish fed diets supplemented with 450 mg of vitamin E showed an increase (P<0.05) in the accumulation of macrophages, foreign body giant cells and Langhans type cells. The kinetics of macrophage recruitment and giant cell formation on the glass coverslips appeared to be strongly influenced by vitamin E supplementation, since fish fed a basal diet and held at high stocking densities showed low numbers of adhering cells on the coverslips, and high concentrations of plasma corticosteroids. On the other hand, fish given a diet supplemented with 450 mg of vitamin E did not show a similar difference in plasma cortisol concentrations related to stocking density. The effect of cortisol concentrations on carbohydrate metabolism, analysed by assessment of plasma glycaemia, was not clear. Blood glucose concentrations did not vary substantially with the different treatments examined. These results suggest that vitamin E may contribute to the efficiency of the fish's inflammatory response by increasing macrophage recruitment and giant cell formation in the foreign body granulomatous reaction. Vitamin E appeared to act on the stress response of pacus by preventing a stress-related immunosuppression. © 2005 Elsevier Ltd. All rights reserved.

Puncture wounds by driftwood catfish during bucket baths: Local habits of riverside people and fish natural history in the Amazon

Objective. - To assess the prevalence of stings by small spiny driftwood catfish (carataí) of the genus Centromochlus (Auchenipteridae) accidentally caught in buckets during bucket bathing by riverside people along the Brazilian Amazon and to determine the probability of catching specimens of these fish during random throws of a bucket into the river. Methods. - We interviewed 27 adult residents living at the confluence of the Negro and Solimões rivers in Brazil regarding whether or not they had ever been stung by driftwood catfish while bucket bathing. To assess the likelihood of catching catfish in bathing buckets, we randomly threw a typical plastic bucket used for bathing in 4 series of 10 throws into the river at dusk or night around a floating house. Results. - Seventeen of the 27 subjects (63%) reported being injured by driftwood catfish during bucket bathing. Three individuals (17.6%) had been injured 2 to 3 times, totaling 23 puncture accidents. All stings occurred at dusk or early night. In the 4 series of 10 bucket throws, we caught 3 driftwood catfish (in 1 series we did not catch any fish). Thus, the chance of catching a driftwood catfish in a single bucket throw at dusk was slightly less than 10%. Conclusions. - The prevalence of stings by driftwood catfish to people bucket bathing in this section of the Brazilian Amazon is high, partly because of the relatively high chances of catching these small catfish during random throws of a bathing bucket into the river.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

Cytogenetic characterization of the silverside fish Odontesthes regia (Humboldt, 1833) (Teleostei: Atheriniformes: Atherinopsidae) from Iquique, Chile

This paper describes the karyotype of Odontesthes regia by means of Giemsa staining, C-banding, to reveal the distribution of the constitutive heterochromatin, and by Ag-staining and fluorescent in situ hybridization (FISH), to locate ribosomal genes (rDNA). The chromosome diploid modal count in the species was 2n = 48. The karyotype is composed of one submetacentric pair (pair 1), 16 subtelocentric pairs (pairs 2 to 17), and 7 acrocentric pairs (pairs 18 to 24). With the exception of pair 1 it was not possible to classify the homologous chromosomes accurately because differences in chromosome size were too slight between adjacent pairs. The distribution of C-banded heterochromatin allowed for a more accurate matching of the majority of chromosomes of the subtelocentric series. Silver staining of metaphase spreads allowed for the identification of Nucleolus Organizer Regions (Ag-NOR) on pair 1. FISH experiments showed that 18S rDNA sequences were located, as expected, in the same chromosome pair identified as the Ag-NOR-bearing one.

Bacterial contamination of the catfish (Cathorops agassizii - Valenciennes, 1839 and Genidens genidens - Agassiz, 1829) sting

This work aimed to study the bacterial contamination in stings of the catfish Genidens genidens and Cathorops agassizii found in the São Vicente estuarine system (São Paulo State, Brazil). For bacteriological analyses, we used fish samples distributed into a group of 50 specimens (25 C. agassizii and 25 G. genidens) and a group of 14 specimens (7 C. agassizii and 7 G. genidens). Results showed contamination of 13 different bacterial species of Enterobacteriaceae, being Klebsiella pneumoniae the most frequent bacteria (26.80%) followed by Enterobacter sp and Escherichia coli (16.27%), and Serratia marcescens, Serratia sp. and Proteus mirabilis (1.16%). Gram-positive bacteria as well as fungi were not detected in the samples. According to the Gram-negative species characterized and with regard to the environmental conditions, it can also be considered that accidents with these catfish stings may develop significant acute secondary infections in humans.