Variability of Bile Baseline Excitation-emission Fluorescence of Two Tropical Freshwater Fish Species.

The quantification of pollutant metabolites in fish bile is an efficient approach to xenobiotic pollution monitoring in freshwaters since these measurements directly address exposure. Fluorescence excitation-emission matrix spectroscopy (EEMS) has demonstrated to be a highly specific and cost-effective technique for polycyclic aromatic hydrocarbon (PAH) and PAH-metabolite identification and quantification. EEMS ability to quantify these compounds strongly depends on the intensity and variability of the bile baseline fluorescence (BBF). We found large differences in BBF among Aequidens metae (AME) individuals and of these with Piaractus orinoquensis (PIO). Moreover, BBF was large enough that solvent dilutions of over 1:400 were needed to avoid inner filter effects. We used parallel factor analysis (PARAFAC) to model the intra- and inter-species BBF variability. PARAFAC successfully decomposed the EEMS set into three fluorophores present in all samples, although in concentrations spreading over ~ 3 orders of magnitude. One of the factors was identified as tryptophan. Tryptophan and Factor 2 were covariant and much more abundant in AME than in PIO, while Factor 3 was ~ 6 times more abundant in PIO than in AME. Also, tryptophan was ~ 10x more abundant in AME specimens immediately caught in rivers than in their laboratory-adapted peers. The PARAFAC decomposition effectiveness was confirmed by the positive proportionality of scores to dilution ratios. A large inner filter indicates that Factor 2 is as strong a light absorber as tryptophan. Our results stress the need to include bile matrix variable components for the detection and quantification of pollutant metabolites using PARAFAC.

Seroprevalence of exposure to SARS-CoV-2 in domestic dogs and cats and its relationship with COVID-19 cases in the city of Villavicencio, Colombia.

Background: Since the beginning of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak, different animal species have been implicated as possible intermediate hosts that could facilitate the transmission of the virus between species. The detection of these hosts has intensified, reporting wild, zoo, farm, and pet animals. The goal of this study was to determine the seroprevalence of anti-SARS-CoV-2 immunoglobulins (IgG) in domestic dogs and cats and its epidemiological association with the frequency of coronavirus disease 2019 (COVID-19) patients in Villavicencio, Colombia. Methods: 300 dogs and 135 cats were randomly selected in a two-stage distribution by clusters according to COVID-19 cases (positive RT-qPCR for SARS-CoV-2) within the human population distributed within the eight communes of Villavicencio. Indirect enzyme-linked immunosorbent assay (ELISA) technique was applied in order to determine anti-SARS-CoV-2 IgG in sera samples. Kernel density estimation was used to compare the prevalence of COVID-19 cases with the seropositivity of dogs and cats. Results: The overall seroprevalence of anti-SARS-CoV-2 IgG was 4.6% (95% CI=3.2-7.4). In canines, 3.67% (95% CI=2.1-6.4) and felines 6.67% (95% CI=3.6-12.18). Kernel density estimation indicated that seropositive cases were concentrated in the southwest region of the city. There was a positive association between SARS-CoV-2 seropositivity in pet animals and their habitat in Commune 2 (adjusted OR=5.84; 95% CI=1.1-30.88). Spearman's correlation coefficients were weakly positive ( p=0.32) between the ratio of COVID-19 cases in November 2020 and the results for domestic dogs and cats from the eight communes of Villavicencio. Conclusions: In the present research cats were more susceptible to SARS-CoV-2 infection than dogs. This study provides the first positive results of anti-SARS-CoV-2 ELISA serological tests in domestic dogs and cats in Colombia with information about the virus transmission dynamics in Latin America during the COVID-19 pandemic.

Liver biomarkers response of the neotropical fish Aequidens metae to environmental stressors associated with the oil industry.

The Acacias River in Colombia receives large volumes of industrial effluents mostly derived from the oil industry. To contribute to the study of the possible effects of industrial wastewaters on the aquatic environment and particularly on fish populations, a native neotropical fish, Aequidens metae was used as a sentinel species. Wild specimens of A. metae were caught at three different places of the Acacias River taking as reference the point of discharge of an oil industry effluent; upstream, downstream, and at the vicinity of the discharge pipe. A fourth sampling site was chosen as a reference site away from urban settlements. Samplings were performed twice, during the rainy and dry seasons. After anesthesia animals were weighted and measured, and humanely sacrificed. Livers were extracted, frozen on site and transported to the laboratory. Condition indices were calculated. Total protein content and the detoxification 7-ethoxyresorufin-O-deethylase (EROD) enzyme activity were estimated. Histopathological alterations were also evaluated. Water quality was estimated through the measurement of several variables. Results obtained evidenced that the highest induction in EROD activity and the strongest histological alterations in liver of the monitored fish appeared during the dry seasons at the discharge site and downstream to this point.

Biochemical responses and proximate analysis of Piaractus brachypomus (Pisces: Characidae) exposed to phenanthrene.

Polycyclic Aromatic Hydrocarbons (PAHs) are complex compounds generated from industrial and anthropogenic activities, although natural phenomena are also responsible for their presence in the environment. The aim of this study was to evaluate the effect of phenanthrene on proximate composition and antioxidant activity in Piaractus brachypomus during a subacute exposure. Fish were exposed intraperitoneally to phenanthrene (0.1, 1.0, and 10 µg g-1) and a solvent control (0 µg g-1). Muscle tissue, liver and bile were collected at 0 h, 11 days and 21 days. In dorsal muscle, parameters such as dry matter, protein, crude fat, and pH did not show alterations with exposure to phenanthrene (p ≥ 0.05). At 11 days, a significant increase of hepatic lipid peroxidation was observed in fish exposed to 10 µg g-1. Additionally, a dose-dependent response was observed in the muscle, although no significant differences were observed in the activity of catalase and lipid peroxidation. Phenanthrene metabolites in bile were analyzed by fixed fluorescence at 260/380 nm (excitation/emission) wavelengths and levels indicated a dose-dependent response. Likewise, bioaccumulation of phenanthrene in bile was observed for 21 days, suggesting slow metabolism of this xenobiotic at 10 µg g-1. This study provides important information on the oxidative effects generated by phenanthrene exposure for 21 days in P. brachypomus, generating fundamental information on adverse effects induced by PAHs on the most important Colombian native fish species. These data contribute to the development of additional research in environments contaminated with PAHs and can be applied to monitor areas contaminated with oil spills.

Biochemical and histological alterations in Aequidens metae (Pisces, Cichlidae) and Astyanax gr. bimaculatus (Pisces, Characidae) as indicators of river pollution.

The water quality from rivers near to cities has decreased drastically over the decades due to the environmental pollution. The Ocoa River on its way through the Villavicencio city, Colombia, receives large volumes of domestic and industrial wastewater. In order to establish the effect of the contamination, biochemical and histopathology biomarkers were evaluated in the gills and the liver of two native fish species. Astyanax gr. bimaculatus and Aequidens metae were caught in three sites of the Ocoa River (S1, S2 and S3) and in a reference River during the rainy and dry season. A. metae showed to be more sensitive to water pollution. In general, the rainy season induced a greater negative impact in the fish monitored. At site S3 (after crossing the city where urban runoff and industrial effluents are discharge and close to a landfill), increased inhibition of the antioxidant response and lipid membranes damage were observed; in addition, in this site the histopathological alterations were greater in both fish species. The impact observed in this study on fish health provoked for the Ocoa River contamination demands the need to begin strategies to solve the problem of discharge of domestic and industrial wastewater in aquatic environments in Colombia.

Physiological and enzymatic responses of Chlorella vulgaris exposed to produced water and its potential for bioremediation.

In South America, Colombia is known as an important oil-producing country. However, the environmental impact of crude oil industry has not been studied deeply and few studies have been carried out for evaluating responses of algae and its adaptation under specific conditions. Enzymatic and physiological effects in Chlorella vulgaris and its potential for bioremediation after exposure to produced water (PW) were assessed using different PW concentrations (0, 25, 50, 75 and 100%) and crude oil. Variables such as cell density, growth rate (µ), percentage of growth inhibition (% I), chlorophyll a and b and cell diameter were evaluated during 5 days. Furthermore, enzymatic biomarkers such as superoxide dismutase (SOD) and catalase (CAT) were also measured. Results showed that the treatment with 100% PW had the highest cell density and µ; similarly, 25% PW treatment had a similar behaviour, being these two treatments with the highest growth. A dose-dependent response was seen for chlorophyll a and b and cell diameter, showing significant differences between treatments and the control. Different levels of SOD and CAT were observed in algae exposed to PW. At 24 h, an increase in SOD and CAT activity was observed, probably due to effects caused by xenobiotics. After 72 h, a decrease in the activity of both enzymes was observed. The results evidenced that C. vulgaris can adapt easily to PW, showing an increase on its growth and stabilisation in its antioxidant activity. Additionally, cell diameter results and decrease of hydrocarbons and phenols show the potential of these algae to degrade xenobiotics from PW.

Hábitos alimenticios de Leporinus friderici (Anostomidae: Teleostei) durante un ciclo hidrobiológico en el río Vaupés, Colombia / Feeding habits of Leporinus friderici (Anostomidae: Teleostei) during a hydrobiological cycle in Vaupés River, Colombia

ResumenLa especie más importante de la pesquería del río Vaupés es un pez conocido como Warakú tres puntos (Leporinus friderici), un carácido migratorio representativo del departamento del Vaupés, Colombia, de importancia para la seguridad alimentaria de comunidades indígenas. Con el propósito de contribuir en el conocimiento biológico básico de la especie, el presente estudio determinó los hábitos y preferencias alimenticias de Leporinus friderici durante un ciclo hidrobiológico (marzo 2014 a mayo 2015) en el río Vaupés, Colombia. Fueron muestreados tres lugares, dos a lo largo del río Vaupés (comunidades indígenas de Yacayacá y Santa Cruz) y uno en el río Cuduyarí afluente del río Vaupés (comunidad de Piracemo). Se analizaron 316 contenidos estomacales aplicando los métodos de frecuencia de ocurrencia (FO), índice gravimétrico (W), coeficiente de vacuidad (CV) e índice de importancia relativa (IIR). De igual modo, se determinaron parámetros físicos y químicos del agua superficial y profunda. La profundidad del río en época de aguas ascendentes fue de 3.7 ± 0.6 m, aguas altas de 5.9 ± 1.4 m, aguas descendentes 4.6 ± 1.3 m y en aguas bajas 2.4 ± 1.0 m; se registró mayor acidez y turbidez del agua en las ascendentes. El CV fue de 14.5 %; sin embargo, el CV alcanzó un valor del 40 % en aguas ascendentes. En general se observó una mayor FO de material vegetal en los estómagos de los individuos muestreados (44.4-66.7 %), seguido por los insectos (21.1-33.3 %, dietas secundarias). Los índices gravimétricos reflejaron que el material vegetal fue el ítem consumido en mayor cantidad, seguido de los insectos y en casi igual proporción invertebrados y material animal. Las diferencias en la composición de la dieta confirman la naturaleza oportunista de esta especie con predominio de hábitos omnívoros.Este estudio puede ser utilizado como parte integral del conocimiento de la ecología trófica de la especie, con el fin de crear estrategias para la protección del L. friderici en la región del Vaupés.

AbstractThe most important fish species of the Vaupes river is known as a Warakú tres puntos (Leporinus friderici), the most representative migratory Characidae from the Vaupés state, Colombia, with a significant importance in food security of indigenous communities. To contribute with the knowledge of its basic biology, we determined the habits and food preferences of Leporinus friderici, during one hydrobiological cycle (March 2014 to May 2015) in the Vaupés river, Colombia. Three sites were sampled, two of them along the Vaupés river (Yacayacá and Santa Cruz communities) and the other one in the Cuduyarí river, tributary of the Vaupés river (Piracemo community). Physical and chemical water parameters from the surface and depth level of the river were determined. Stomach contents from 316 fish were analyzed by frequency of occurrence (FO), gravimetric index, vacuity index (VI) and relative importance index (RII). Vaupés's average depth in ascending water period was of 3.7 ± 0.6 m, during rainy season of 5.9 ± 1.4 m, in descending water period of 4.6 ± 1.3 m and during dry season of 2.4 ± 1.0 m. The higher acidity and turbidity were observed during the ascending water period. The average VI was 14.5 % for the period, but it reached 40 % in descending waters. In general, a FO of plant material was the most frequent (44.4-66.7 %), followed by insects (21.1-33.3 %, secondary diets). The gravimetric indices showed that plant material was the most consumed item, followed by insects, and in almost in equal proportion, invertebrates and animal material. The differences in diet composition confirm the opportunistic nature of this species with a predominance of omnivorous habits. This study can be used as an integral part of the feeding ecology of L. friderici knowledge, in order to develop strategies for its protection in the Vaupés region because of its importance for local communities.

Using genotoxic and haematological biomarkers as an evidence of environmental contamination in the Ocoa River native fish, Villavicencio-Meta, Colombia.

The studies evaluating the impact of environmental pollution on aquatic organisms have had a growing concern in recent decades. The Ocoa River runs through Villavicencio city, receiving the wastewater produced in the city. To date studies reporting the effect and the consequences of Ocoa River pollution on aquatic organisms inhabiting this water body are unknown. Haematological and genotoxicity biomarkers were evaluated in peripheral blood of two native fish species Astyanax gr. bimaculatus and Aequidens metae during rainy and dry season at three different sites of Ocoa River called Nacimiento (site 1, before entering the city), Centauros (site 2, inside the city), Caño Seco (site 3, after the city). Also, fish from a reference site with little likelihood of contamination were sampled. Alterations such as decrease in erythrocyte count, lymphocytes, haemoglobin concentration and haematocrit percentage, and increased thrombocytes and neutrophils counts, together with nuclear abnormalities in erythrocytes such as increased frequency of micronuclei, lobed, blebbed and notched nuclei and binucleate cells were observed in fish from the site 3, followed by the site 2 in rainy season. These results shown that during rainy season, sites 2 and 3 receive domestic and industrial wastewater leading to genotoxic and haematological changes in the monitored organisms.

Fecundidad y fertilidad de Macrobrachium amazonicum (Héller 1862) (Decápoda, Palaemonidae) del Piedemonte Llanero Colombiano / Fecundity and fertility of Macrobrachium amazonicum (Héller 1862) (Decápoda, Palaemonidae) of Colombian Piedemonte Llanero

Objetivo. Determinar la fecundidad y fertilidad de Macrobrachium amazonicum del Piedemonte llanero. Materiales y métodos. Los animales fueron capturados en una granja piscícola del municipio de Restrepo-Meta, Colombia y llevados a la estación piscícola del Instituto de Acuicultura, Universidad de los Llanos. Se utilizaron hembras ovígeras a las cuales se les determinó el peso corporal (g) y longitud total (mm), así como su fecundidad (número de huevos/ hembra) y fertilidad (número de larvas/hembra). En los huevos se midieron el diámetro y longitud, para calcular su volumen. Resultados. La fecundidad varió entre 120 a 549 huevos con una media de 331 ± 93. Con respecto a la fertilidad, las hembras evaluadas tuvieron 287 ± 19 larvas, con un intervalo entre 102 a 703. Tanto para fecundidad como para fertilidad se observó que existe una relación lineal positiva significativa (r2 entre 0.85 y 0.91) con respecto al peso corporal y la longitud total de las hembras (p< 0.001). El diámetro de los huevos fue de 750.7 ± 2.9 μm (rango: 500 – 1000 μm) y el largo 974.2 ± 3.6 μm (525 – 1200 μm). El tamaño de los huevos fue de 296.2 ± 9.7 μm3. Conclusiones. Los valores de la fecundidad y fertilidad para Macrobrachium amazonicum en el Piedemonte llanero son inferiores a los resultados obtenidos en otros estudios; sin embargo, el tamaño de sus huevos es mayor. Los resultados permiten inferir que la especie se reproduce durante todo el año y que su fácil manejo permitiría su cultivo en cautiverio.

Objective. To determine the fecundity and fertility of Macrobrachium amazonicum (freshwater shrimp) of the Piedemonte llanero. Materials and methods. The animals were caught at a fish farm, in the municipality of Restrepo, Meta, Colombia and transported to the fish station of the Institute of Aquaculture of the University of the Llanos. The body weight (g), and total length (mm) of mature females and the fecundity and fertility (number of eggs and larvae/female, respectively) were determined. The diameter and the length of the eggs were determined to calculate their volume. Results. The fecundity varied from 120 to 549 eggs with an average of 331 ± 93. Regarding fertility, the females showed 287 ± 19 larvae, with an interval between 102 and 703. A highly positive lineal regression (r2 between 0.85 and 0.91) was observed between fecundity and fertility, and body weight and total length (p < 0.001). The egg diameter was 750.7 ± 2.9 µm (range: 500 - 1000 µm) and egg length of 974.2 ± 3.6 µm (525-1200 µm). The egg size was 296.2 ± 9.7 µm3. Conclusions. The determination of fecundity and fertility for Macrobrachium amazonicum in the Piedemonte Llanero was lower to the results obtained in other studies; however, the size of the eggs was higher. The results could infer that the species breeds throughout the year and allows easy handling and farming in captivity.

Effect of culture medium on growth and protein content of Chlorella vulgaris / Efecto del medio de cultivo sobre el crecimiento y el contenido proteico de Chlorella vulgaris / Efeito do diferentes meios de cultura sobre o crescimento e o conteúdo proteico em Chlorella vulgaris

The use of live food in larviculture and fingerling stages has become an excellent option in pisciculture to reduce mortality and production costs. Live food is cost-effective, nutritious, and it has enzymes necessary for fish development. Objective: this study evaluated the effect of several culture media (chu10 , NPK complex fertilizer, worm humus, and equine manure) on growth and protein content of Chlorella vulgaris. Methods: 3 L volumes with constant light and aeration were used for algae culture. Six replicates were used per each treatment. One mL of C. vulgaris was added to each experimental unit to obtain 8.3 x 10(6) cel/mL as initial density. Algae counts were conducted every two days using the Neubauer chamber to determine cell density, parameters such as temperature, dissolved oxygen, pH, nitrates, and phosphates were measured. Crude protein was determined with the Kjeldahl method. The assay lasted 82 days. Results: microalgae reached the highest density with complex fertilizer NPK (10.9 ± 1.6 x 10(6) cel/mL) on day 22, followed by worm humus (5.3 ± 1.1 x 10(6) cel/mL) on day 48, equine manure (4.9 ± 0.9 x 10(6) cel/mL) on day 18, and the last was chu10 (2.2 ± 0.6 x 10(6) cel/mL) on day 12. The highest protein content was found in algae grown in worm humus (56.8%), and the equine manure (32.5%), in contrast, complex fertilizer NPK had the lowest value (16.8%). Conclusion: this study showed that organic media are a good choice for the culture of C. vulgaris, providing proper growth and high protein content of microalgae.

En piscicultura, el uso de alimento vivo durante las fases de larvicultura y alevinaje se ha convertido en una opción para reducir la tasa de mortalidad y los costos de producción, debido a que es económico, nutritivo y posee enzimas necesarias para el desarrollo de las larvas de peces. Objetivo: el presente trabajo evaluó el efecto de diferentes medios de cultivo (chu10, fertilizante complejo NPK, humus de lombriz y equinaza) sobre el crecimiento y el contenido proteico en Chlorella vulgaris. Métodos: para su cultivo se utilizaron volúmenes de 3 L, con luz y aireación constante. De cada tratamiento se realizaron seis réplicas y en cada unidad experimental se adicionó 1 mL de C. vulgaris con una densidad de 8.3 x 10(6) cel/mL. Para determinar la densidad celular se realizaron conteos cada dos días haciendo uso de cámara de Neubauer. Se midió temperatura, oxígeno disuelto, pH, nitratos y fosfatos. La proteína cruda se determinó por el método de Kjeldahl. La duración total del ensayo fue de 82 días. Resultados: la microalga alcanzó su mayor densidad con fertilizante complejo NPK (10.9 ± 1.6 x 10(6) cel/mL) en el día 22, seguido del humus de lombriz (5.3 x 106 ± 1.1 x 10(6) cel/mL) en el día 48, equinaza (4.9 ± 0.9 x 10(6) cel/mL) en el día 18, y por último con el chu10 (2.2 ± 0.6 x 10(6) cel/mL) en el día 12. Se encontró mayor contenido proteico en las células cultivadas en humus de lombriz (56.8%) y equinaza (32.5%); a diferencia, el del fertilizante complejo NPK fue muy bajo (16.8%). Conclusiones: este estudio permite concluir que los medios orgánicos son una buena opción para cultivar C. vulgaris, otorgando un adecuado crecimiento y un alto contenido proteico de la microalga.

Na piscicultura, o uso de alimento vivo durante as fases da cultura de larvas e juvenis têm sido convertidos em uma opção para reduzir a taxa de mortalidade e os custos de produção, porque é econômico, nutritivo e possui as enzimas necessárias para o desenvolvimento das larvas de peixe. Objetivo: o objetivo desta pesquisa foi avaliar a efeito de diferentes meios de cultura (chu10, fertilizante mineral complexo NPK, vermicomposto e esterco equino) sobre o crescimento e o conteúdo proteico em Chlorella vulgaris. Métodos: para a cultura, foram utilizados volumes de 3 L, com luz e aeração permanente. De cada tratamento, realizaram-se seis repetições e em cada unidade experimental, adicionou-se 1 mL de C. vulgaris com uma densidade de 8.3 x 10(6) cel/mL. Para determinar a densidade celular, realizaram-se contagens a cada dois dias usando a câmara de Neubauer. Aferiu-se a temperatura, oxigênio dissolvido, pH, nitratos e fosfatos. A proteína bruta foi determinada por o método de Kjeldahl. A duração total do ensaio foi de 82 dias. Resultados: os resultados mostraram que a microalga alcançou sua maior densidade com o fertilizante complexo NPK (10.9 ± 1.6 x 10(6) cel/mL) no dia 22, seguido do vermicomposto (5.3 ± 1.1 x 10(6) cel/mL) no dia 48, o esterco equino (4.9 ± 0.9 x 10(6) cel/mL) no dia 18 e por último com o chu10 (2.2 ± 0.6 x 10(6) cel/mL) no dia 12. Foi encontrado maior conteúdo proteico nas células cultivadas em vermicomposto (56.8%) e esterco equino (32.5%); a diferença, do fertilizante complexo NPK que foi muito baixo (16.8%). Conclusões: este estudo permitiu concluir que os meios orgânicos são uma boa opção para cultivar C. vulgaris, obtendo um crescimento adequado e um alto conteúdo proteico da microalga.

Requerimientos nutricionales de peces ornamentales de agua dulce: una revisión / Nutritional requirements of freshwater ornamental fish: a review

The knowledge of nutritional requirements in ornamental fish species is essential to improve the productive development; however, the nutritional information of these species is scarce and sometimes this information is extrapolated from results obtained from non-ornamental fish species. In ornamental fish, a correct formulation of the diet improve the nutrient digestibility and supply the metabolic needs, reducing the maintenance cost and at the same time the water pollution. Inert food such as meal powder, flakes, milk powder, bovine heart and liver, tubifex worms, as well as live food including Artemia sp., rotifers and Moina have been used extensively in ornamental fish feeding with a diverse range of nutritional values and productive properties. In contrast with farmed fish, skin pigmentation is a mandatory characteristic in ornamental fish and the use of dietary supplements with carotenoids is recommended. The aim of this document is to review the specific nutritional requirements which are indispensable to improve economical and productive potential of freshwater ornamental fish.

Bezafibrate, a lipid-lowering pharmaceutical, as a potential endocrine disruptor in male zebrafish (Danio rerio).

Fibrates are pharmaceuticals commonly used to control hypercholesterolemia in humans and they are frequently detected in the freshwater environment. Since cholesterol is the precursor of all steroid hormones, it is suspected that low cholesterol levels will impact steroidogenesis. However, the effect of fibrates on fish reproductive endocrinology is not clear; therefore the aim of the present study was to evaluate the effect of bezafibrate (BZF) on gonadal steroidogenesis and spermatogenesis of zebrafish (Danio rerio). For this purpose, adult males were exposed orally to 1.7, 33 and 70 mg BZF/g food for 21 days. Blood and gonads were collected after 48 h, 7 days and 21 days to evaluate plasma cholesterol and plasma 11-ketotestosterone (11-KT). The expression of gonadal genes involved in the steroidogenesis was quantified to determine a potential mechanism of action, likewise the effect on spermatogenesis was evaluated by examining gonadal histopathology. A time dependent monotonic decrease in the plasma cholesterol concentration was observed in fish exposed to BZF. Plasma 11-KT decreased significantly after 21 days of exposure in fish exposed to the high concentration of BZF. Different gene expression patterns were observed: down-regulation in ppara and pparg mRNA levels was observed in fish exposed to the higher concentrations after 48 h; however, the expression of pparg increased after 21 days. After 21 days an increase in the star and cyp17a1 mRNA expression was observed in fish exposed to 70 mg BZF/g food. Sampling time and bezafibrate concentration explained 52.4% and 20%, respectively, of the gene expression variability. Gonadal histology revealed the presence of germ cell syncytia in the tubular lumen of fish exposed to bezafibrate and also an increased number of cysts containing spermatocytes, which indicate testicular degeneration. The study shows that bezafibrate exerts a hypocholesterolemic effect in adult male zebrafish and its potential as an endocrine disruptor due to its effect on the gonadal steroidogenesis and spermatogenesis.

Endosulfan affects health variables in adult zebrafish (Danio rerio) and induces alterations in larvae development.

Adult zebrafish (Danio rerio) were exposed to 0 (control), 0.16 or 0.48µg/L of the insecticide, endosulfan, for 28days. Haematology, whole body ions, thiobarbituric acid reactive substances (TBARS), Na(+)K(+)-ATPase, organ histology and reproduction were assessed in adults. The resulting offspring were examined for latent effects on development (heart rate and morphometrics). On day 14, adult fish exposed to 0.16µg/L endosulfan showed significantly lower red blood cell counts than those exposed to 0.48µg/L endosulfan; adult fish exposed to 0.16 ug/L also showed elevated TBARS compared to controls. Both concentrations of endosulfan caused a 4.0 fold increase in Na(+)K(+)-ATPase activity compared to controls (ANOVA, p<0.05). On day 14, the livers of fish exposed to endosulfan had fewer, enlarged hepatocytes, with cell diameters greater than the controls (ANOVA, p<0.05). Morphological alterations in the progeny of fish exposed to endosulfan were observed. Heart beat frequency was significantly lower in larvae from exposed adults to 0.16 µg/L compared to the control (ANOVA, p<0.05). These findings show that sublethal exposure to endosulfan causes adverse sublethal effects in adult D. rerio, and effects on the development of their offspring.

EFICIENCIA ANÉSTESICA DE 2-FENOXIETANOL, BENZOCAINA, QUINALDINA Y METASULFONATO DE TRICAINA EN ALEVINOS Y JUVENILES DE CACHAMA BLANCA (Piaractus brachypomus) / EFFICIENCY OF 2-PHENOXYETHANOL, BENZOCAINE, QUINALDINE AND TRICAINE METHASULPHONATE AS AN ANESTHESIA FOR Pirapitinga (Piaractus brachypomus) FINGERLINGS AND JUVENILE FISHES

Objetivo. Evaluar la efectividad anestésica de 2-fenoxietanol, benzocaina, quinaldina y metanosulfonato de tricaina (MS-222) en alevinos y juveniles de cachama blanca (Piaractus brachypomus). Materiales y métodos. Alevinos (7.3 ± 6.8 g) y juveniles (49.6 ± 28.4 g) fueron expuestos a 2-fenoxietanol (200, 400 y 600 ppm), benzocaina (50, 100 y 150 ppm), quinaldina (7.5, 15 y 30 ppm) o MS-222 (100, 150 y 200 ppm) (n=12), para evaluar la duración de la inducción y recuperación, frecuencia opercular y la sobrevivencia postanestesia. Resultados. El desarrollo corporal y la concentración afectaron el periodo de inducción, siendo mayor en juveniles que en alevinos (p<0.05) y menor a medida que aumentó la concentración de anestésico; sin embargo, alevinos expuestos a MS-222 mostraron períodos de inducción más largos que los juveniles; y alevinos expuestos a 2-fenoxietanol mostraron períodos de inducción iguales con las tres concentraciones (p>0.05), mientras que juveniles expuestos a MS-222 mostraron el menor tiempo de inducción con la menor concentración (p<0.05). El tiempo de recuperación fue inferior a 2 min, con 2-fenoxietanol, MS-222, quinaldina y 50 ppm de benzocaina; pero altas concentraciones de benzocaina mostraron largos períodos de recuperación. No se observó tendencia en la variación de la frecuencia opercular ni mortalidad durante o después de la exposición a los anestésicos. Conclusiones. En alevinos el 2- fenoxietanol (400 ppm) y en juveniles el MS-222 (100 ppm), serían las sustancias de elección para procedimientos cortos, pero deberá recurrirse a altas concentraciones de benzocaina cuando el propósito de la anestesia sea un procedimiento prolongado.

Objective. To evaluate the efficiency of 2-phenoxyethanol, benzocaine, quinaldine and tricaine methasulphonate (MS-222) as an anesthesia for pirapitinga (Piaractus brachypomus) fingerlings and juvenile fishes. Materials and methods. Fingerlings (7.3 ± 6.8 g) and juveniles (49.6 ± 28.4 g) were exposed to 2-phenoxyethanol (200, 400 and 600 ppm), benzocaine (50, 100 y 150 ppm), quinaldine (7.5, 15 y 30 ppm) or MS-222 (100, 150 y 200 ppm) (n=12) in order to evaluate the induction and recovery time, opercular frequency and post-anesthesia survival. Results. Induction time was longer in juveniles than in fingerlings (p<0.05) and decreased in proportion to the increase in the concentration of anesthesia. However, induction time in fingerlings exposed to MS-222 was longer than in juveniles. Fingerlings exposed to 2-phenoxyethanol had similar induction times at the three concentrations (p>0.05), whereas the juveniles exposed to the low MS-222 concentration had lower induction time (p<0.05). The recovery time was less than 2 min. using 2- phenoxyethanol, MS-222, quinaldine and 50 ppm of benzocaine, but was longer with higher benzocaine concentrations. No variation in opercular frequency or mortality was observed during or after anesthesia. Conclusions. In fingerlings and juveniles, 2-phenoxyethanol (400 ppm) and MS-222 (100 ppm), respectively, are considered the anesthetics of choice for short procedures. Higher concentrations of benzocaine should be used for longer procedures.

Behavioural and gill histopathological effects of acute exposure to sodium chloride in moneda (Metynnis orinocensis).

To evaluate the toxicity of sodium chloride (NaCl), juveniles and adult Metynnis orinocensis were exposed for 96h to 0, 5, 10, 15, 20 or 40gL(-1) of salt. Food intake, behaviour, opercular frequency (OF), mortality, body weight and gill microscopic alterations were evaluated. Behavioural changes were observed in fish exposed to concentrations higher than 10gL(-1). Juveniles and adults showed a progressive decrease in the OF and body weight. Food intake decreased in concentrations below 15gL(-1). Juveniles and adults exposed to 15, 20 or 40gL(-1) had 100% mortality. Lamellar congestion, hyperplasia and fusion were the common microscopic alterations at higher concentrations. The gill congestion severity increased with salt concentration. The LC(50) for juveniles and adults were 10.5gL(-1) and 10.8gL(-1), respectively. These results suggest that salt concentrations lower than 5gL(-1) are safe for preventive and therapeutic practices in Metynnis orinocensis; whereas prolonged exposure higher than 10gL(-1) is deleterious in this species.

Determinación del espermatocrito y efecto del volumen de la dosis seminante sobre la fertilidad en yamú (Brycon amazonicus) / Determination of spermatocrit and effect of volume of insemination dose on fertility in yamú (Brycon amazonicus)

El yamú (Brycon amazonicus . Spix & Agassiz 1829) es un pez omnívoro de rápido crecimiento y carne de alta calidad y aceptación comercial. Para estandarizar el espermatocrito (SPCTO), 23 muestras de semen fueron centrifugadas a 14000 g durante 1, 3, 5, 7 ó 10 min. El tiempo de centrifugación tuvo un efecto significativo sobre el valor del espermatocrito (SPCTO) (p<0.05). Para conocer la concentración espermática (CE) durante la época reproductiva y su relación con el SPCTO, la espermiación de 40 machos fue inducida con extracto de hipófisis de carpa (4.0 mg.kg -1). La CE fue evaluada mediante cámara de Neubauer (dilución 1:1200) y el SPCTO por centrifugación a 14000 g durante 5 min. La relación entre SPCTO y CE fue positiva y altamente significativa (y= 1.69 + 0.59x; r2= 0.74; p<0.0001), pero no se observaron diferencias estadísticas (p>0.05) entre los meses evaluados (marzo, abril y mayo). Finalmente, para determinar el efecto del volumen de la dosis seminante sobre la fertilidad, 2 g de oocitos fueron seminados con 3, 15, 45, 90 o 180 mL de semen (n= 12), cuya CE fue previamente ajustada para contener 50000 espermatozoides (sptz)/oocito, utilizando como diluyente plasma seminal homólogo. Los porcentajes de fertilidad fueron 83, 86, 86, 85 y 87 por ciento, respectivamente (p>0.05).

Protocolo para la crioconservación de semen de yamú (Brycon amazonicus Spix & Agassiz 1829) / Protocol for cryopreservation of yamú (Brycon amazonicus) semen

La crioconservación es una herramienta importante para el almacenamiento de gametos por tiempo indefinido, especialmente de especies con hábitats y poblaciones amenazadas. En peces, es además una alternativa para optimizar los procesos reproductivos en cautiverio de especies con maduración gonadal asincrónica y ciclos reproductivos estacionales como el yamú (Brycon amazonicus - Spix & Agassiz 1829), un carácido distribuido en las cuencas de los ríos Amazonas, Orinoco y Essequibo. Este reporte presenta el protocolo para la crioconservación de semen en esta especie, obtenido después de evaluar diferentes sustancias crioprotectoras, curvas de congelación y volúmenes de empaque. Su eficiencia ha sido demostrada en varios experimentos a nivel de laboratorio y de granjas comerciales, lográndose tasas de fertilidad cercanas a aquellas con semen fresco, constituyéndose en una importante herramienta metodológica para la crioconservación seminal de la especie.

Evaluación de diferentes crioprotectores para la crioconservación de espermatozoides de yamú (Brycon amazonicus) / Evaluation of different cryoprotectors for cryopreservation of yamú (Brycon amazonicus) spermatozoa

El uso de dimetilsulfoxido (DMSO), etilenglicol (ETG), propilenglicol (PPG) y metanol (MET) para la crioconservación de semen de yamú (Brycon amazonicus - Spix & Agassiz 1829) fue evaluadocon el objetivo de determinar la calidad post-descongelación y la fertilidad in vitro del semen. Para este propósito, machos y hembras, sexualmente maduros fueron inducidos a la maduración final de lasgónadas por medio de extracto de hipófisis de carpa. El semen obtenido fue evaluado y diluido (1:4) en una solución a base de yema de huevo (12 por ciento) y glucosa (5.5 por ciento). Se utilizó cada crioprotector aconcentraciones de: 3.75, 5, 7.5, 10, 11.25 y 15 por ciento para un total de 24 tratamientos. El semen diluido fue empacado en pajillas de 0.5 mL para ser congelado en vapores de nitrógeno líquido (NL) a -194 ºC durante 30 min y luego transferidas y almacenadas hasta por 10 días en un termo de NL para su evaluación. Para las pruebas de fertilidad, las pajillas (n= 6) de cada uno de los tratamientos fueron descongeladas a 35 ºC por 60 seg y posteriormente evaluadas. Se encontró que los tratamientos DMSO-5 por ciento, ETG-3.75 por ciento, PPG-3.75 por ciento y MET-3.75 por ciento presentaron los mayores porcentajes de movilidad (PM) y tiempos de activación (TA), siendo DMSO-5 por ciento el más alto (PM 76 ± 2.4; TA 88.2 ± 6.1). De otro lado, los tratamientos PPG-3.75 por ciento y MET-3.75 por ciento mostraron los mayores porcentajes de fertilidad (50 ± 1.4 y 47.8 ± 1.4, respectivamente), pero fueron inferiores (p<0.05) cuando se compararon con el semen fresco (64.7 ± 1.7). En general se observó que a medida que aumentó la concentración del crioprotector, la calidad seminal postdescongelación disminuyó, sin observarse relación directa entre la movilidad espermática y la fertilidad.

Efectos del sistema de conservación sobre la fertilidad de oocitos de yamú (Brycon amazonicus) durante cortos períodos de almacenamiento / Effects of the conservation system on eggs fertility of yamú (Brycon amazonicus) during shortterm storage

El objetivo fue evaluar los efectos del sistema de conservación sobre la fertilidad de oocitos de yamú (Brycon amazonicus) durante cortos períodos de almacenamiento. Se utilizaron hembras y machosadultos tratados con extracto de hipófisis de carpa para estimular la ovulación y aumentar el volumen seminal. Oocitos obtenidos de la misma hembra fueron sometidos a tres sistemas de conservación: in situ (IS, mantenidos dentro de la cavidad ovárica), ex situ a temperatura ambiente (T°A) y ex situ a temperatura de refrigeración (T°R). El diámetro ovocitario (DO) y la viabilidad fueron evaluados a los 0 (control), 15, 30, 60 y 120 min de almacenamiento. El DO fue medido con reglilla acoplada al ocular del estéreomicroscopio y la viabilidad mediante prueba de fertilidad, seminando 3 g de oocitos con 250 mL de semen fresco. También fueron medidos la longitud total y el volumen del saco vitelino de las larvas. Bajo los tres sistemas de conservación, DO aumentó con el tiempo, siendo mayor (p<0.001) que el control a partir de los 15 min de almacenamiento. Los mayores DO fueron observados en oocitos conservados a T°R (p<0.05). Hasta 60 min de almacenamiento, en los sistemas de conservación IS y a T°A, la fertilidad no varió (p>0.05) con respecto al control. A los 120 min de almacenamiento, todos los sistemas de conservación mostraron fertilidad menor que el control; sin embargo, en el sistema T°R disminuyó más rápidamente, siendo menor (p<0.05) que el control a los 30 min. Aunque las larvas no presentaron evidentes alteraciones morfológicas, las del control fueron más largas (p<0.01) que aquellas de oocitos conservados. En conclusión, oocitos ovulados de yamú pueden mantenerse viables hasta por 60 min, conservándolos en la cavidad ovárica o a temperatura ambiente. En contraste, la refrigeración deterioró rápidamente la fertilidad de oocitos ovulados.

Efecto del peso corporal sobre el consumo de oxígeno en yamú (Brycon amazonicus Spix & Agassiz 1829): reporte preliminar / Effects of the body weight on the routine oxygen consumption of yamú (Brycon amazonicus): preliminary report

Con el propósito de evaluar los efectos del peso corporal sobre el consumo de oxígeno en yamú (Brycon amazonicus), ejemplares de 100, 200, 400, 800 y 1600 g de peso corporal fueron alojados en un respirómetro de 172 L de capacidad y registrado el consumo de oxígeno cada cinco minutos durante dos a cuatro horas, utilizando una sonda multiparamétrica YSI 556. La temperatura del agua se mantuvo a 28 ± 1 °C. Los resultados mostraron una relación inversa entre la tasa de consumo de oxígeno y el peso corporal, ya que los animales de menor talla registraron un mayor consumo de oxígeno por unidad de peso que aquellos de pesos superiores. Esta relación generó la siguiente ecuación de regresión: Y= 583.7 - 64.9 Ln(X), donde Y es el consumo de oxígeno en mg/kg/h y Ln(X) el logaritmo natural del peso corporal expresado en gramos (r2= 0.92, n= 5).