Anesthesia by sprinkling method in the gills of tambaqui Colossoma macropomum does not influence intensity and morphology of monogeneans / Anestesia por meio de aspersão nas brânquias de tambaqui Colossoma maropomum não influencia a intensidade e a morfologia de monogeneas

Abstract The present study evaluates the influence of anesthesia on the parasitic fauna of monogenea fish parasites, as its intensity and viability. Two experiments were conducted: Evaluation of an anesthetic method by sprinkling eugenol directly on gills and evaluation of monogenea motility and viability; Comparison of immersion and directly sprinkling on the gills with benzocaine and eugenol followed by evaluation on parasite intensity. The results suggest that the anesthetic sprinkling didn't interfere in the parasite motility, morphology and body surface integrity analyzed by fluorescence method. The monogenean intensity in the gills was lower in fish anesthetized by immersion method compared to the sprinkling method and the control group. This method of anesthesia can be used in parasitological studies.

Resumo O presente estudo avalia a influência da anestesia sobre a fauna parasitária de monogeneas em peixes, sua intensidade e sua viabilidade. Dois experimentos foram realizados: Avaliação de um método anestésico por aspersão eugenol diretamente nas brânquias e avaliação da motilidade das monogeneas e sua viabilidade; e Comparação entre imersão e aspersão diretamente nas brânquias com benzocaína e eugenol, seguido de avaliação sobre a intensidade parasitária. Os resultados sugerem que a aspersão do anestésico não interferiu na motilidade, morfologia, superfície corporal e integridade do parasita, analisadas pelo método de fluorescência. A intensidade de monogenéticos nas brânquias foi menor nos peixes anestesiados pelo método de imersão em comparação com o método de aspersão e o grupo controle. O método de anestesia por aspersão nas brânquias pode ser utilizado em estudos parasitológicos.

Expression studies on NA+/K+-ATPase in gills of Penaeus monodon (Fabricius) acclimated to different salinities.

The decapod crustacean Penaeus monodon survives large fluctuations in salinity through osmoregulation in which Na+/K+-ATPase (NKA) activity in the gills plays a central role. Adult P. monodon specimens were gradually acclimatized to 5, 25 and 35‰ salinities and maintained for 20 days to observe long term alterations in NKA expression. Specific NKA activity assayed in gill tissues was found to be 3 folds higher at 5‰ compared to 25‰ (isosmotic salinity) and 0.48 folds lower at 35‰. The enzyme was immunolocalized in gills using mouse α-5 monoclonal antibody that cross reacts with P. monodon NKA α-subunit. At 5‰ the immunopositive cells were distributed on lamellar tips and basal lamellar epithelium of the secondary gill filaments and their number was visibly higher. At both 25‰ and 35‰ NKA positive cells were observed in the inter-lamellar region but the expression was more pronounced at 25‰. Gill architecture was normal at all salinities. However, the 1.5 fold increase in NKA α-subunit mRNA at 5‰ measured by quantitative RT-PCR (qRT-PCR) using EF1α as reference gene was not statistically significant. The study confirms the osmoregulating ability of P. monodon like other crustaceans at lower salinities. It is likely that significant increase in NKA transcript level happens at an earlier time point. At higher salinities all three methods record only marginal or no change from isosmotic controls confirming the hypothesis that the animal largely osmoconforms in hyperosmotic environment.

Carbaryl-induced alterations in biochemical metabolism of the prawn, Macrobrachium malcolmsonii.

The present study was performed to investigate the toxic impact of carbaryl on biochemical metabolism in the hemolymph, brain, hepatopancreas, gills and muscle of intermoult juveniles of the economically important prawn, Macrobrachium malcolmsonii. The concentration of glutathione S-transferase (GST) was found to be higher in test prawns when compared with controls. This suggests that a mechanism of detoxification was in operation to neutralise carbaryl toxicity. However, the toxic effect of carbaryl was not fully neutralised, and hence, alterations were recorded in basic biochemical metabolism of test prawns. The concentration of acetylcholinestrase (AchE) was found to be lower in test prawns than that of controls. Carbaryl toxicity resulted in utilisation of major biochemical constituents, such as total carbohydrate, glycogen, protein and lipid to generate required energy as an attempt to withstand the toxic stress. Glycogenolysis resulted in elevation of total free sugar level in the hemolymph of test prawns. While proteolysis led to elevation of total free amino acid level in test prawns. The content of total lipid have also been found lower in test prawns than that of controls. This suggests that carbaryl toxicity resulted in severe energy crises in test prawns. In the present study, toxic effects of carbaryl impair basic metabolic functions and hence pose a threat to the life of M. malcolmsonii.

The ultrastructure of the gastrodermis and the nutrition of the gill parasitic Atriaster heterodus Lebedev and Paruchin, 1969 (Platyhelminthes: Monogenea)

The gastrodermis of Atriaster heterodus Lebedev & Paruchin, 1969 (Polyopisthocotylea), a gill parasite from Diplodus argenteus (Valenciennes, 1830), is composed of "U"-shape hematin cells and a connecting syncytium, both having cytoplasmic lamellae. These cells show outgrowths and bent folds which were seen to enclose lumen material. The trapped material was then subjected to endocytosis. The nature of ingested food material was comparatively analysed by cytochemical and histochemical tests. Blood residues were detected in the gut but tests for mucins were negative. No intact erythrocytes were observed in the gut lumen.

The interaction between O2 and CO2 movements during aerobic exercise in fish

In resting rainbow trout, 99 per cent of the total CO2 excreted across the gills consisted of bicarbonate (HCO3-) dehydrated to CO2 in the red cell. This value decreased to 93 percent during exercise, the remainder being excreted as molecular CO2 which existed in the pre-branchial blood. HCO3- dehydration consumes a proton which can be supplied from hemoglobin (1-1b) through its buffer capacity or the Haldane effect. Bohr proton release from Hb upon oxygenation is maximal between 50 and 100 percent of Hb saturation, consistent with observations made in tench blood. At low swimming speeds, when venous blood 02 content (CvO2) was high, there was a small but insignificant acidosis as blood passed through the gills, indicating a greater release of protons than could be consumed by HCO3-. At higher swimming speeds, when CvO2 was low, there was a significant alkalosis in the arterial blood relative to the venous blood, indicating that fewer protons were released upon oxygenation than HCO3- ions were dehydrated to CO2. The disproportionate release of Bohr protons over the range of Hb-O2 saturation in the blood at the gills limits HCO3- dehydration during greater work loads, conserving the HCO3- buffer capacity of the blood and tissues.

Serotonergic vasomotor control in fish gills

Serotonin (5-hydroxytryptamine, 5-HT)-containing neuroepithelial cells (NECs) have been discovered in the gills of fish belonging to different phylogenetic groups, and an additional serotonergic innervation of the gill filaments is present in teleosts. The most apparent effect of serotonin is a bronchial vasoconstriction. For teleosts, it has been postulated that the serotonergic innervation of the proximal portion of the efferent filamental artery (EFA) and adjacent efferent lamellar arterioles (ELA) acts to constrict the vasculature. Thus, as perfusion pressure increases, more lamellae are recruited. In contrast, recent results from rainbow trout show that one precise action of serotonin is a vasoconstriction on the distal portion of the filament vasculature. This may explain why exogenously added serotonin impairs gas exchange in rainbow trout. In addition to constricting the respiratory (arterio-arterial) pathway, injection of serotonin into the Atlantic cod dilates the arterio-venous pathway, diverting blood flow to the arterio-venous pathway. The vasoconstrictory effects of serotonin can be blocked by the 5-HT-receptor antagonist methysergide, whereas the vasodilatory effects cannot.

Gill diffusion as a physiological mechanism for hydrogen peroxide elimination by fish

Hydrogen peroxide is metabolized by the specific enzymatic action of catalase and glutathione peroxidase in animal tissues. The relatively low catalase and glutathione peroxidase activities found in the blood of fish may be related to the ability of gills to eliminate hydrogen peroxide into the aquatic environment. Poecilia vellifera releases hydrogen peroxide apparently by gill diffusion into the environment, resulting in a steady-state H2O2 concentration of about 0.6 µM in the surrounding water. This physiological mechanism resembles ammonia excretion by teleost fish