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Atmospheric particulate matter (APM) produced by the steel industry comprises a complex mixture of particles that includes a wide variety of metals and metallic nanoparticles. These particles settle out onto areas surrounding the industries. There is evidence that this 'settleable' APM (SePM) may cause air-to-water cross-contamination with significant effects on aquatic biota. Recent investigations have reported sublethal impacts on the gill structure and blood oxygen-carrying capacity of fishes, which raises the hypothesis that there will be consequences for gas exchange capacity and ability to support aerobic activities. Therefore, we investigated the effects of an environmentally relevant level of SePM contamination on swimming performance and associated aerobic metabolic rates in Nile tilapia, Oreochromis niloticus. Short-term exposure (96 h) to SePM reduced critical swimming speed, energetic efficiency of aerobic swimming, standard metabolic rate, maximum metabolic rate, and aerobic scope. The compromised swimming performance could have adverse ecological effects by limiting foraging ability, predator evasion, territorial protection, and migration. The impairments to aerobic capacity could also affect overall fish performance by influencing long-term energy balance and allocation to growth and reproduction. Thus, despite being sublethal, SePM contamination is considerably debilitating, and if its limiting effects are not compensated for in the longer term, this may reduce the survival and fitness of fish populations.

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This study investigated the activity of lactated dehydrogenase (LDH), malate dehydrogenase (MDH) enzymes and the levels of glucose, protein and triglyceride in bullfrog tadpoles after exposure to 1 µg L-1 of zinc (Zn), copper (Cu) and cadmium (Cd) isolated and combined for 2 and 16 days. Zn, Cu + Cd and Zn + Cu + Cd increased the activity of the LDH (2 and 16 days) and MDH (2 days) enzymes in the liver; and MDH increased in the kidney after 16 days in all co-exposed groups compared to the control. Glucose increased in the liver in the Zn and Cu groups at 2 and 16 days of exposure and decreased in the kidney (groups Cd, Zn + Cd and Cu + Cd) and muscle (Cd) at 2 days of exposure. After 2 days of exposure, the protein increased in the liver (Zn), in the kidney in all groups exposed to metals except in the groups exposed to Cd and Zn + Cu + Cd, which did not change and decreased in muscle in all the groups exposed to isolated metals. Regarding triglycerides, the kidney and muscle were the most affected, leading to a decrease in the Zn, Cu and Cd groups and in the Zn + Cu (16 days) and Zn + Cu + Cd groups (2 days). The anaerobiosis and aerobiosis were activated in the liver and kidney after short-term exposure (2 days) and in the kidney, the aerobic metabolism was activated after chronic exposure (16 days). The metals caused toxicity and were higher in co-exposure to metals with a potential to cause metabolism damage in L. catesbeianus.

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The gill structure of the Amazonian fish Arapaima gigas (Cuvier 1829) shows ontogenetic changes during development, particularly due the transition from the aquatic to the obligatory air breathing mode of respiration. However, three main cell types can be found in the gills: mitochondrial rich cells, pavement cells and mucous cells (MCs). The MCs are involved in the secretory pathway. The functions of the secreted molecules include mechanical protection of epithelia, protection against parasites and bacterial infection, and role on ion regulation. In this study, we analysed mucous cell location and mucous cell type, based on pH, during the development of A. gigas. Using samples obtained from the environment, gills were collected and fixed in buffered solution. Histological techniques for the identification of MCs were performed Alcian Blue (AB) and periodic acid-Schiff (PAS). The results showed the presence of PAS+ and AB+ cells in the whole filament in all examined fish. In animals less than 50 g, few MCs were present, and no differences were observed in AB+ and PAS+ cells. In animals weighing close to 500 g, more PAS+ cells than AB+ cells were observed, and in animals that weighed more than 1,000 g, more AB+ cells than PAS+ cells were observed. These observations may be a result of the ontogenetic changes in the gill epithelia, which can change the osmorespiratory compromise in ion regulation functions as well the glycosaminoglycans secreted by PAS cells, which in large animals can play a role in the protection against parasites and bacterial infection.

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Radiocystis fernandoi R28 strain is a cyanobacterium which produces mostly the RR and YR microcystin variants (MC-RR and MC-YR, respectively). The effects of crude extract of the R. fernandoi strain R28 were evaluated on the protein phosphatases and on the structure and ultrastructure of the liver of the Neotropical fish, Hoplias malabaricus, after acute and subchronic exposure. Concomitantly, the accumulation of the majority of MCs was determined in the liver and muscle. The fish were exposed to 120.60 MC-RR+MC-LR kg-fish-1 (=100µg MC-LReq kg-fish-1) for 12 and 96h (one single dose, acute exposure) and 30days (one similar dose every 72h, subchronic exposure). MCs did not accumulate in the muscle but, in the liver, MC-YR accumulated after acute exposure and MC-RR and MC-YR accumulation occurred after subchronic exposure. Protein phosphatase 2A (PP2A) activity was inhibited only after subchronic exposure. Acute exposure induced liver hyperemia, hemorrhage, changes in hepatocytes and cord-like disorganization. At the ultrastructural level, the decreasing of glycogen and lipid levels, the swelling of mitochondria and whirling of endoplasmic reticulum suggested hepatocyte necrosis. Subchronic exposure resulted in a complete disarrangement of cord-like hepatocytes, some recovery of mitochondria and whirling endoplasmic reticulum and extensive connective tissues containing fibrous materials in the liver parenchyma. Despite microcystin toxicity and liver alterations, no tumor was induced by MCs. In conclusion, the increased algal mass of R. fernandoi in tropical freshwater, producing mainly MC-RR and MC-YR variants, results in fish liver impairments.

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Application of sewage sludge in agricultural lands is a growing practice in several countries due to its numerous benefits to soil and crops, where chemical and pathogen levels are determined by corresponding legislation. However, the presence of contaminants in residues must always be controlled before application due to their dangerous effects over the ecosystem and potential risks to human health. The main objective of this study was to integrate biological and chemical analysis in order to help elucidating the residue potential toxic, cytotoxic, and mutagenic effects. We evaluate samples of sewage sludge before and after the sanitizing treatment with lime in cytokinesis-block assay using CHO-k1 culture cells. The sanitizing treatment promoted a decrease in pathogen levels, which is the main purpose of this process. Even with chemical levels below the established by environmental agencies, results showed sewage sludge ability to enhance genotoxic and mutagenic effects, proving that residue should be handled with caution in order to minimize its environmental and human risk.

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The present study investigated the structural changes in the rat calcaneal tendon (CT), superficial flexor tendon (SFT), and deep flexor tendon (DFT) in response to jump exercises and anabolic androgenic steroids (AAS). Animals were divided into four groups: sedentary, trained, AAS-treated sedentary rats, and AAS-treated trained animals. Training increased the volume density (Vv%) of blood vessels in all regions of the CT and DFT, cell Vv% in the peritendinous sheath of the proximal and distal regions of the SFT and proximal region of DFT, and cell Vv% in the tendon proper of the proximal and distal regions of the SFT and DFT. The combination of AAS and load exercises showed little increased blood vessel Vv% at the proximal region of the CT, intermediate region of the SFT, and all regions of the DFT as opposed to an increase in adipose cell Vv% in the CT proximal region. The AAS reduced the levels of hydroxyproline in the proximal region of the DFT and in the distal region of the STF. In conclusion, exercise promoted benefits to the adaptation of the tendons to overload. These effects were absent when load exercise was combined with AAS. The abusive consumption of AAS contributes to tendon inertness and rigidity, and increases the potential risk of injury.

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Avaliaram-se os efeitos dos níveis de suplementação de colina em dietas para frangos de corte, de 22 a 40 dias de idade, sobre o desempenho, a composição de carcaças e fígados, bem como as características macroscópicas e histopatológicas dos fígados. O período de criação foi de um a 40 dias de idade, sendo que o período experimental foi de 22 a 40 dias de idade. Até o 21º dia de idade, todas as aves receberam a mesma ração à qual foram adicionados 400mg colina/kg. No 22º dia, os frangos foram distribuídos em cinco tratamentos, nos quais os níveis testados de suplementação de colina foram: zero, 400, 800, 1.200 e 1.600mg/kg, com seis repetições de 30 aves cada. As rações experimentais, formuladas à base de milho, farelo de soja e farinha de carne/ossos, foram isonutritivas, com exceção dos níveis de colina. O delineamento experimental foi o inteiramente ao acaso, e a estimativa da suplementação de colina foi avaliada pelos modelos de regressão. Os níveis de suplementação de colina apresentaram efeito linear pelo teste F (P<0,05) para o consumo de ração. Os demais parâmetros avaliados não sofreram influência da suplementação (P>0,05). O nível de 1.600mg colina/kg de ração resulta em maior consumo de ração de frangos de corte aos 40 dias de idade.

The effects of choline supplemental levels were evaluated in diets for broilers on performance, carcass and liver composition and liver histopathology characteristics from 22 to 40 days-old. The period of creation was one to 40 days of age, and the experimental period was 22 to 40 days-old. Until 21 days of age all birds received the same diet added to 400mg choline/kg. On day 22, broilers were allotted to five treatments (zero, 400, 800, 1200 and 1600 mg/kg), with six repetitions of 30 birds each. The experimental diets formulated based on corn, soybean meal and meat/bones meal were isonutritive, except for the choline levels. The experimental design was completely randomized and the estimation of choline supplementation was evaluated by regression models. The levels of choline supplementation showed a significant linear effect for the F test (P<0.05) for feed intake. The other evaluated parameters were not influenced by supplementation (P>0.05). The supplementation of 1600mg choline/kg in the diet results in higher feed intake of broilers at 40 days old.

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Avaliaram-se os efeitos dos níveis de suplementação de colina em dietas para frangos de corte, de 22 a 40 dias de idade, sobre o desempenho, a composição de carcaças e fígados, bem como as características macroscópicas e histopatológicas dos fígados. O período de criação foi de um a 40 dias de idade, sendo que o período experimental foi de 22 a 40 dias de idade. Até o 21º dia de idade, todas as aves receberam a mesma ração à qual foram adicionados 400mg colina/kg. No 22º dia, os frangos foram distribuídos em cinco tratamentos, nos quais os níveis testados de suplementação de colina foram: zero, 400, 800, 1.200 e 1.600mg/kg, com seis repetições de 30 aves cada. As rações experimentais, formuladas à base de milho, farelo de soja e farinha de carne/ossos, foram isonutritivas, com exceção dos níveis de colina. O delineamento experimental foi o inteiramente ao acaso, e a estimativa da suplementação de colina foi avaliada pelos modelos de regressão. Os níveis de suplementação de colina apresentaram efeito linear pelo teste F (P<0,05) para o consumo de ração. Os demais parâmetros avaliados não sofreram influência da suplementação (P>0,05). O nível de 1.600mg colina/kg de ração resulta em maior consumo de ração de frangos de corte aos 40 dias de idade.(AU)

The effects of choline supplemental levels were evaluated in diets for broilers on performance, carcass and liver composition and liver histopathology characteristics from 22 to 40 days-old. The period of creation was one to 40 days of age, and the experimental period was 22 to 40 days-old. Until 21 days of age all birds received the same diet added to 400mg choline/kg. On day 22, broilers were allotted to five treatments (zero, 400, 800, 1200 and 1600 mg/kg), with six repetitions of 30 birds each. The experimental diets formulated based on corn, soybean meal and meat/bones meal were isonutritive, except for the choline levels. The experimental design was completely randomized and the estimation of choline supplementation was evaluated by regression models. The levels of choline supplementation showed a significant linear effect for the F test (P<0.05) for feed intake. The other evaluated parameters were not influenced by supplementation (P>0.05). The supplementation of 1600mg choline/kg in the diet results in higher feed intake of broilers at 40 days old.(AU)

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The effects of oviposition time, which represents different positions of the follicles in a sequence of posture, were studied on egg weight, chick weight, egg weight loss during incubation, hatchability, embryo mortality and infertility . A total of 19, 820 eggs, layed on the same day, from Cobb® broiler breeders aging 46-week-old were used. The eggs were classified according to the oviposition time posture (corresponding to the time of collection) from 8:30 am, 10:30 am, 12:30 pm, 14:30 pm and 16:30 pm, which represented the folowing treatments: 1st, 2nd, 3rd, 4th and 5th collects, respectively. The experimental design was completely randomized with five treatments and 36 repetitions, and each tray of 96 eggs was considered a repetition. The normal and homogeneous data were submitted to analysis of variance and means compared by Tukey test. The non-normal and non-homogeneous averages were compared by Kruskal-Wallis test. It is concluded that the time of egg laying has influence on egg weight and chick weight, but does not affect hatchability, fertility and embryonic development.

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The effects of oviposition time, which represents different positions of the follicles in a sequence of posture, were studied on egg weight, chick weight, egg weight loss during incubation, hatchability, embryo mortality and infertility . A total of 19, 820 eggs, layed on the same day, from Cobb® broiler breeders aging 46-week-old were used. The eggs were classified according to the oviposition time posture (corresponding to the time of collection) from 8:30 am, 10:30 am, 12:30 pm, 14:30 pm and 16:30 pm, which represented the folowing treatments: 1st, 2nd, 3rd, 4th and 5th collects, respectively. The experimental design was completely randomized with five treatments and 36 repetitions, and each tray of 96 eggs was considered a repetition. The normal and homogeneous data were submitted to analysis of variance and means compared by Tukey test. The non-normal and non-homogeneous averages were compared by Kruskal-Wallis test. It is concluded that the time of egg laying has influence on egg weight and chick weight, but does not affect hatchability, fertility and embryonic development.(AU)

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O presente experimento foi conduzido em lote de matrizes Cobb desde 46 até 60 semanas de idade. Os tratamentos foram definidos pelas dietas (com e sem adição de 6ppm de cantaxantina na dieta das matrizes) e pelos períodos de armazenamento dos ovos (três e sete dias), em delineamento inteiramente ao acaso, em arranjo fatorial 2x2 (duas dietas x dois períodos de armazenamento). A cantaxantina dietética elevou o escore de cor da gema de sete para 14. O armazenamento dos ovos por sete dias prejudicou a qualidade dos ovos, promoveu oxidação de ácidos graxos da gema e retardou o desenvolvimento embrionário.(AU)

This trial was done with Cobb broiler breeders. Treatments were defined by the diets (with and without 6ppm canthaxanthin added to broiler breeder diets) and periods of egg storage (three and seven days) in a completely randomized factorial 2 x 2 design (two diets x two periods of egg storage). The dietary canthaxanthin increased the yolk color score from 7 to 14. Egg storage for seven days reduced egg quality, promoted oxidation of yolk fatty acids and delayed embryonic development.(AU)

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O presente experimento foi conduzido em lote de matrizes Cobb desde 46 até 60 semanas de idade. Os tratamentos foram definidos pelas dietas (com e sem adição de 6ppm de cantaxantina na dieta das matrizes) e pelos períodos de armazenamento dos ovos (três e sete dias), em delineamento inteiramente ao acaso, em arranjo fatorial 2x2 (duas dietas x dois períodos de armazenamento). A cantaxantina dietética elevou o escore de cor da gema de sete para 14. O armazenamento dos ovos por sete dias prejudicou a qualidade dos ovos, promoveu oxidação de ácidos graxos da gema e retardou o desenvolvimento embrionário.

This trial was done with Cobb broiler breeders. Treatments were defined by the diets (with and without 6ppm canthaxanthin added to broiler breeder diets) and periods of egg storage (three and seven days) in a completely randomized factorial 2 x 2 design (two diets x two periods of egg storage). The dietary canthaxanthin increased the yolk color score from 7 to 14. Egg storage for seven days reduced egg quality, promoted oxidation of yolk fatty acids and delayed embryonic development.

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This study investigated the relationship between contaminant body burden and the oxidative stress status of the gills and livers of two wild fish species in the Furnas Hydroelectric Power Station (HPS) reservoir (Minas Gerais, Brazil). Gills and livers presented similar pathways of metals and organochlorine bioaccumulation. During June, organochlorines were associated with lipid peroxidation (LPO), indicating oxidative stress due to the inhibition of the antioxidant enzymes superoxide dismutase and glutathione peroxidase. In the most polluted areas, metal concentrations in the liver were associated with metallothionein. During December, contaminants in the gills and liver were associated with catalase activity and LPO. Aldrin/dieldrin was the contaminant most associated with oxidative damage in the livers of both species. This integrated approach shed light on the relationship between adverse biological effects and bioaccumulation of contaminants inputted by intensive agricultural practices and proved to be a suitable tool for assessing the environmental quality of man-made reservoirs.

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Through integrating chemical, biochemical and morphological analyses, this study investigated the effects of multiple pollutants on the gill mitochondria-rich cells (MRCs) in two fish species, Astyanax fasciatus and Pimelodus maculatus, collected from five sites (FU10, FU20, FU30, FU40 and FU50) in the Furnas Hydroelectric Power Station reservoir. Water analyses revealed aluminum, iron and zinc as well as organochlorine (aldrin/dieldrin, endosulfan, heptachlor/heptachlor epoxide and metolachlor) contamination at all of the sites, with the exception of FU10. Copper, chrome, iron and zinc were detected in the gills of both species, and aldrin/dieldrin, endosulfan and heptachlor/heptachlor epoxide were detected in the gills of fish from all of the sites, with the exception of FU10. Fish collected at FU20, FU30 and FU50 exhibited numerous alterations in the surface architecture of their pavement cells and MRCs. The surface MRC density and MRC fractional area were lower in fish from FU20, FU30, FU40 and FU50 than in those from the reference site (FU10) in the winter, and some variability between the sites was observed in the summer. The organochlorine contamination at FU20 and FU50 was associated with variable changes in the MRCs and inhibition of Na(+)/K(+)-ATPase (NKA) activity, especially in P. maculatus. At FU30, the alterations in the MRCs were associated with the contaminants present, especially metals. A multivariate analysis demonstrated a positive association between the biological responses of both species and environmental contamination, indicating that under realistic conditions, a mixture of organochlorines and metals affected the MRCs by inhibiting NKA activity and inducing morphological changes, which may cause an ionic imbalance.

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The impact of acute (48 h) and subchronic (14 days) exposures to environmentally realistic atrazine concentrations (2, 10 and 25 µg L(-1)) were evaluated on the gills of Prochilodus lineatus by assessing the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), the levels of reduced glutathione (GSH) and lipid peroxide (LPO) as well as the histopathological damage. Acute and subchronic exposure to atrazine at 2 or 25 µg L(-1) did not change the activities of GST, SOD, CAT or GPx or the concentrations of GSH and LPO; however, subchronic exposure to 10 µg L(-1) increased the activity of GST, SOD and CAT and the LPO level. Histopathological indexes indicated normal gill function with scattered epithelial changes after acute and chronic exposure to 2 or 10 µg L(-1) of atrazine; however, fish chronically exposed to 25 µg L(-1) of atrazine, although had scattered lesions, the severity of lesions resulted in slightly to moderately gill damage. Acute exposure to atrazine decreased the type 3 MCs (containing acid mucosubstances with sulfate esters) in fish exposed to 2 or 10 µg L(-1) and increased the type 4 MCs (containing all types of mucosubstances) in fish exposed to 25 µg L(-1). Chronic exposure to atrazine reduced the type 3 MCs in fish exposed to 10 or 25 µg L(-1). The gills showed a low sensitivity to atrazine after acute exposure. However, the persistence of atrazine in water (subchronic exposure) promoted an increase of LPO levels in the gills and increased the frequency and severity of histopathological changes. The decreased density of type 3 MCs in fish exposed to atrazine suggests a mechanism to wash toxic substances away from the gill surface.

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The effects of the herbicide atrazine on the gill of the freshwater fish Prochilodus lineatus were evaluated after exposure of fish to 2, 10 and 25 µg L(-1) atrazine during 48 h (acute exposure) and 14 d (subchronic exposure). Ions and osmolality were measured in plasma and gill samples were taken to determine the Na(+)/K(+)-ATPase (NKA) and carbonic anhydrase (CA) activities and for morphological analysis. Plasma osmolality and Na(+) and Cl(-) ions changed depending on atrazine concentration, but atrazine exposure had no effect on the Na(+)/Cl(-) ratio. NKA activity did not change after atrazine exposure, but CA activity decreased in fish exposed to 25 µg L(-1) for 14 d. Gill MRC density decreased after acute exposure but did not change in fish exposed to the subchronic treatment. The MRC density at the epithelial surface increased in fish exposed to 25 µg L(-1), and the MRC fractional area (MRCFA) increased in fish exposed to 10 µg L(-1). The changes in MRCs provide evidence of morphological adjustments to maintain ionic homeostasis in spite of the inhibition of CA activity at the highest atrazine concentration.

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In order to verify the influence of chronic and acute ambient oxygen levels from egg to adult stage of the zebrafish, in vivo oxygen consumption (MO2), critical tensions of oxygen (Pcrit), heart rate (fH) and total body lactate concentration (Lc) were determined for Danio rerio (Hamilton, 1822) raised at 28 degrees C under normoxic (7.5 mgO2.L-1 or 80 mm.Hg-1) and hypoxic conditions (4.3 mgO2.L-1) and exposed to acute hypoxia during different developmental stages. Our findings confirmed that very early stages do not respond effectively to ambient acute hypoxia. However, after the stage corresponding to the age of 30 days, D. rerio was able to respond to acute hypoxia through effective physiological mechanisms involving aerobic and anaerobic metabolism. Such responses were more efficient for the fishes reared under hypoxia which showed that D. rerio survival capability increased during acclimation to mild hypoxia. Measurements of body mass and length showed that moderate hypoxia did not affect growth significantly until the fish reached the stage of 60 days. Moreover, a growth delay was verified for the hypoxic-reared animals. Also, the D. rerio eggs-to-larvae survival varied from 87.7 to 62.4% in animals reared under normoxia and mild hypoxia, respectively. However, the surviving animals raised under moderated hypoxia showed a better aptitude to regulate aerobic and anaerobic capacities when exposed to acute hypoxia.

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Deformation of biological tissues may occur during histological processing and results in loss of accuracy when quantitative information about cells, tissues and organs is necessary. In this study, the gill tissue from armored catfish (Pterygoplichthys anisitsi) was quantified in each step of processing using the stereological principles. During processing for glycol methacrylate embedding, gill tissue from shrinks significantly but regains its original dimensions after sectioning.

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Deformation of biological tissues may occur during histological processing and results in loss of accuracy when quantitative information about cells, tissues and organs is necessary. In this study, the gill tissue from armored catfish (Pterygoplichthys anisitsi) was quantified in each step of processing using the stereological principles. During processing for glycol methacrylate embedding, gill tissue from shrinks significantly but regains its original dimensions after sectioning.

Deformações nos tecidos podem ocorrer durante o processamento histológico e resultar em informações errôneas quando há necessidade de dados quantitativos sobre células, tecidos e órgãos. Neste estudo, o tecido branquial do cascudo (Pterygoplichthys anisitsi) foi quantificado em cada etapa do processamento utilizando os princípios de estereologia. O tecido branquial reduziu significativamente durante processamento histológico com metacrilato, mas retornou às suas dimensões iniciais depois de seccionadas, o que indica não ocorrer nenhuma perda na informação quantitativa do tecido.

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During low-water period, freshwater stingray Paratrygon aiereba collected in the whitewater (WW) of the River Amazon showed higher urea content, osmolality, Na(+) and Cl(-) concentrations in plasma and perivisceral fluid than those caught in blackwater (BW) of the River Negro. Gills and kidney Na(+)-K(+)-ATPase activities were significantly lower in WW than in BW fish. The high level of kidney Na(+)-K(+)-ATPase activity in P. aiereba may minimize ion loss and generate diluted solute-free urine in ion-poor BW environment.

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