A new species of Huffmanela (Nematoda: Trichosomoididae) parasitizing Colomesus psittacus (Tetraodontiformes) from Marajó Island, Pará State, Brazil / Uma nova espécie de Huffmanela (Nematoda: Trichosomoididae) parasitando Colomesus psittacus (Tetraodontiformes) da Ilha de Marajó, Pará, Brasil

Abstract The puffer fish Colomesus psittacus, is not commercialized on Marajó Island. They are captured as bycacth and discarded dead in the environment in artisanal fisheries that occur in the estuaries of northern Brazil. In this sense, the objective was to identify the parasites present in the gills and to evaluate the histopathological alterations caused by these nematodes of the genus Huffmanela. Fifty-five fish were analyzed, and thirty-five specimens showed the parasite in the gills. Morphological characteristics suggest that it is a new species of the genus Huffmanela, and the histopathological exams showed an edematous inflammation in the secondary lamella and the presence of eggs of this nematode, which is the first record of this parasite in C. psittacus in Brazil.

Resumo O baiacu amazônico Colomesus psittacus é um peixe de importância comercial para a aquariofilia, além de ser capturado como bycacth e descartado morto no ambiente nas pescarias artesanais que ocorrem nos estuários da região Norte do Brasil. Nesse sentido, objetivou-se identificar e descrever nematódeos do gênero Huffmanela em baiacus oriundos da ilha de Marajó, bem como as alterações histopatológicas causadas por eles. Cinquenta e cinco peixes foram analisados, e trinta e cinco espécimes apresentaram o parasita nas branquias. Características morfológicas sugerem tratar-se de uma nova espécie do gênero Huffmanela, e os exames histopatológicos demonstraram uma inflamação edematosa na lamela secundária e presença de ovos desse nematódeo. Esse fato se torna o primeiro registro desse gênero parasito em C. psittacus no Brasil.

Biochemical and molecular impacts of glyphosate-based herbicide on the gills of common carp.

Glyphosate (GLY)-based herbicide, one of the most widely used herbicides, might cause a series of environmental problems and pose a toxicological risk to aquatic organisms. However, data on the potential hazard and toxicity mechanism of GLY to fish gills are relatively scarce. In this study, a subacute toxicity test of common carp (Cyprinus carpio L.) treated with commercial GLY at 52.08 and 104.15 mg L-1 for 7 d was conducted. The results revealed that GLY exposure significantly inhibited Na+/K+-ATPase and increased AST and ALT activities in the fish gills. The biochemical assays results revealed that GLY treatment remarkably altered the transcriptional levels of HSP70 and HSP90; inhibited the activities of SOD, CAT, GPx, GR, and T-AOC; reduced the contents of GSH, but remarkably promoted MDA and PC contents, suggesting that GLY exposure induced oxidative stress and lipids and proteins damage in the carp gills. Further research revealed that GLY exposure also promoted expression of NF-κB, iNOS, IL-1ß, IL-6, IL-8, and TNF-α; altered the levels of IL-10 and TGF-ß, indicating that GLY exposure induced inflammatory response in the fish gills. Additionally, we found that GLY exposure activated apaf-1 and bax and inhibited bcl-2, induced caspase-9 and caspase-3 expression and caused remarkable histological damage in the fish gills. These results may further enriches the toxicity mechanistic theory of GLY to fish gills, which may be useful for the risk assessment of GLY and aquatic organism protection.

Histological evaluation of vital organs of the livebearer Jenynsia multidentata (Jenyns, 1842) exposed to glyphosate: A comparative analysis of Roundup® formulations.

Roundup formulations are herbicides whose active principle is glyphosate. However, these formulations are potentially more toxic to non-target organisms than pure glyphosate. This study aimed to evaluate and compare the toxic potential of the Roundup formulations through histological alterations in fish. Thus, males and females of the neotropical fish species Jenynsia multidentata (Jenyns, 1842) were exposed for 24 or 96 h to the Roundup Original® (RO), Roundup Transorb® (RT) or Roundup WG® (RWG) formulations, at a fixed concentration of 0.5 mg/L of glyphosate. This concentration is close to the maximum glyphosate limits found in the environment and is non-lethal to J. multidentata. The three formulations caused histological damage to the liver, gills and brain of J. multidentata, which increased over the exposure time. Differences in the histological alterations between females and males were observed in the liver and brain. Females were more tolerant to RO and RT than RWG. Males did not exhibit these differences in sensitiveness with formulations. The RWG caused more damage in the liver and gills and RT in the brain. Overall, there were differences in the toxicity of RO, RT and RWG and the toxic effect was presented through histological damage, reinforcing the usefulness of histological biomarkers for Roundup® toxicity. The comparison of the toxic potential of glyphosate-based herbicides is important because it could give support to the governmental organizations to set protective rules for the water ecosystems and human health, as well as to reduce the use of highly toxic formulations in agriculture.

Acute lion's mane jellyfish, Cyanea capillata (Cnideria: Scyphozoa), exposure to Atlantic salmon (Salmo salar L.).

Jellyfish-induced gill pathology relies upon occasional diagnostic observations yet the extent and impact of jellyfish blooms on aquaculture may be significant. Idiopathic gill lesions are often observed in apparently healthy fish. This study exposed Atlantic salmon (Salmo salar L.) smolts to macerated Cyanea capillata at 2.5 and 5 g/L for 2 hr under controlled laboratory conditions. Blood chemistry and gill histopathology were examined over a subsequent 4-week period. Fish showed an acute response to the presence of jellyfish, including characteristic external "whiplash" discoloration of the skin and acute increases in blood electrolytes and CO2 concentration; however, these were resolved within 4 days after exposure. Histopathologically, gills showed first an acute oedema with epithelial separation followed by focal haemorrhage and thrombus formation, and then progressive inflammatory epithelial hyperplasia that progressively resolved over the 4 weeks post-exposure. Results were consistent with the envenomation of gills with cytotoxic neurotoxins and haemolysins known to be produced by C. capillata. This study suggests that many focal hyperplastic lesions on gills, especially those involving focal thrombi, may be the result of jellyfish stings. Thus, the presence of jellyfish and their impact may be severe and understated in terms of marine fish aquaculture and fish welfare.

Proliferation, survival and cell death in fish gills remodeling: From injury to recovery.

Pollutants found dispersed in water can cause irritations on the gills, challenge the immune system and prejudice the welfare of the fish. Here we investigated molecules linked to proliferation, survival, and cell death, as well as inflammatory and vascular control, in a model of fish gill remodeling, from injury to recovery. We assessed the gill histology and immunohistochemistry for PCNA, iNOS, HSP70, and Bax in Hypostomus francisci obtained from a river subjected to chronic anthropic influences and then after they were placed in water of good quality. A total of 30 H. francisci adult individuals were collected and distributed into two groups: euthanized on the day of capture (group 1) and maintained for 30 days in an aquarium (group 2). In all the fish from group 1, the primary and secondary lamellae showed hypertrophy of the respiratory epithelium, lamellar fusion, lifting of the epithelium, aneurysm, hyperemia, and vascular congestion. On the other hand, in all the fish from group 2, restoration of gill integrity was observed, and the primary and secondary lamellae showed a simple epithelium, absence of lamellar fusion, hypertrophy, and aneurysm. Gills of fish from group 1 had higher frequency of cells immunopositive for PCNA, iNOS, HSP70, and Bax than those of fish from group 2 (p < 0.05). The molecular and cellular mechanisms from injury to recovery were proposed, with a balance between survival and cell death signals being essential for determining the gill structure. In addition, the findings indicate that recovery of the structural organization of gills is possible if fishes are maintained in good-quality water, indicating the importance of the conservation of aquatic environments.

Physiological responses and changes in gene expression in the large yellow croaker Larimichthys crocea following exposure to hypoxia.

Organisms at all levels of evolutionary complexity react to hypoxic stress. To clarify the effects of acute hypoxia on physiological and biochemical responses of Larimichthys crocea, we measured the activity levels of the antioxidant enzymes superoxide dismutase and catalase, hemoglobin concentration, functional indices of the liver (aspartate transaminase, alanine transaminase), heart (phosphocreatine kinase), and immune system (alkaline phosphatase), as well as mRNA expression levels of the immunity-related genes Hsp70 and HIF-1α at different time points of hypoxic. In addition, liver, gill, and kidney samples were histologically analyzed. We found that hemoglobin concentration and all enzyme activities increased during hypoxia, although these effects were transient and most indices returned to basal levels thereafter. The extent of the increase in the parameter values was inversely proportional to the dissolved oxygen content. Hsp70 and HIF-1α mRNA expression levels increased significantly in the blood, liver, gills, and kidneys following exposure to hypoxia, which may play an important role in protecting fish against oxidative damage. However, we found histological evidence of hypoxia-induced injuries to the gills, liver, and kidneys, which are involved in breathing, detoxification, and osmotic balance maintenance, respectively. Thus, despite the upregulation of defensive mechanisms, acute hypoxia still caused irreversible damage of organs. In conclusion, we observed that, in response to acute hypoxic stress, L. crocea enhances immune defensive function and antioxidant capacity. A better understanding of the regulation of the molecular anti-hypoxia mechanisms can help speeding up the selective breeding of hypoxia-tolerant L. crocea.

Biomarcadores histopatológicos na espécie Hoplias malabaricus (Pisces, Osteichthyes, Erythrinidae) em uma Unidade de Conservação de São Luís (MA) / Histopathological biomarkers in Hoplias malabaricus (Pisces, Osteichthyes, Erythrinidae) species in a protected area of São Luís (MA)

Neste estudo objetivou-se validar dois órgãos (brânquia e fígado) de Hoplias malabaricus para análise de biomarcadores de contaminação aquática na Área de Proteção Ambiental (APA) do Maracanã em São Luís-MA. Exemplares de traíra foram capturados em uma lagoa de policultivo e no Rio Ambude, localizados na APA do Maracanã. A biometria foi realizada em laboratório. Brânquias e o fígado de cada espécime foram fixados em formol a 10% e mantidos em álcool a 70% até a execução da técnica histológica usual. Cortes de aproximadamente 5µm de espessura foram corados em hematoxilina e eosina (HE). Os resultados referentes à biometria dos peixes capturados durante o período de estiagem indicaram que o comprimento total e padrão dos exemplares foram maiores quando comparados com os indivíduos do período chuvoso. As alterações hepáticas identificadas foram: lipidose, necrose, infiltração leucocitária, vacuolização do citoplasma, hemossiderina e centro de melanomacrófagos. Constataram-se as seguintes lesões branquiais: fusão lamelar, aneurisma lamelar, deslocamento e necrose do epitélio, proliferações de células do muco e dilatação capilar. As respostas biológicas de H. malabaricus, registradas em nível branquial e hepático, podem ser caracterizadas como biomarcadores de contaminação aquática. As brânquias mostraram-se como os melhores órgãos para esse tipo de análise na espécie. Esses resultados indicam que os peixes da região apresentam a saúde comprometida por poluentes presentes nos ambientes analisados...

The aim of the present work was to validate two organs (liver and gill) of Hoplias malabaricus for analysis of aquatic contamination biomarkers in the Maracanã Environmental Protected Area (APA) in São Luís-MA. Trahira animals were captured in a pond polyculture and Ambude River located in Maracanã APA. Biometric analysis was performed in the laboratory. Gills and liver of each specimen were fixed in 10% formalin and kept in 70% alcohol to running the usual histological technique. Cuts of approximately 5μm thick were stained with hematoxylin and eosin (HE). The results concerning the biometrics of the fish caught during the dry season indicated that the total and standard length were higher when compared to the rainy season. The liver changes identified were: lipidosis, necrosis, leukocyte infiltration, vacuolization of the cytoplasm, hemosiderin and melanomacrophages centres. We found the following gill lesions: melting lamellar, lamellar aneurysm displacement, necrosis of the epithelium, proliferation of mucous cells and capillary dilation. The biological responses of H. malabaricus, recorded in gill and liver level, can be characterized as biomarkers of aquatic contamination. The gills appeared as the best organs for this kind of analysis in the species. These results indicate that the fish have health problems caused by pollutants present in the analyzed environments...

Gill damage to Atlantic salmon (Salmo salar) caused by the common jellyfish (Aurelia aurita) under experimental challenge.

BACKGROUND: Over recent decades jellyfish have caused fish kill events and recurrent gill problems in marine-farmed salmonids. Common jellyfish (Aurelia spp.) are among the most cosmopolitan jellyfish species in the oceans, with populations increasing in many coastal areas. The negative interaction between jellyfish and fish in aquaculture remains a poorly studied area of science. Thus, a recent fish mortality event in Ireland, involving Aurelia aurita, spurred an investigation into the effects of this jellyfish on marine-farmed salmon. METHODOLOGY/PRINCIPAL FINDINGS: To address the in vivo impact of the common jellyfish (A. aurita) on salmonids, we exposed Atlantic salmon (Salmo salar) smolts to macerated A. aurita for 10 hrs under experimental challenge. Gill tissues of control and experimental treatment groups were scored with a system that rated the damage between 0 and 21 using a range of primary and secondary parameters. Our results revealed that A. aurita rapidly and extensively damaged the gills of S. salar, with the pathogenesis of the disorder progressing even after the jellyfish were removed. After only 2 hrs of exposure, significant multi-focal damage to gill tissues was apparent. The nature and extent of the damage increased up to 48 hrs from the start of the challenge. Although the gills remained extensively damaged at 3 wks from the start of the challenge trial, shortening of the gill lamellae and organisation of the cells indicated an attempt to repair the damage suffered. CONCLUSIONS: Our findings clearly demonstrate that A. aurita can cause severe gill problems in marine-farmed fish. With aquaculture predicted to expand worldwide and evidence suggesting that jellyfish populations are increasing in some areas, this threat to aquaculture is of rising concern as significant losses due to jellyfish could be expected to increase in the future.

Size effects of suspended particles on gill damage in green-lipped mussel Perna viridis.

The green-lipped mussels Perna viridis were exposed to <500 microm suspended solids (SS) with concentrations of 0 (control), 250, 500, 750 and 1000 mg/L for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significantly higher damages to the cilia on the frontal surface of the gill filaments than that on the abfrontal surface in both demibranchs. Percent ciliary depletion varied with SS concentrations and time. No sign of recovery of the gill filaments was observed after the mussels were transferred to clean seawater. In a second experiment, mussels were exposed to SS with size range from <63, >125-<250 and >250-<500 microm at 600 mg/L, together with a control (0 mg/L) for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significant ciliary damages in both the ascending and descending lamellae under the three particle size groups as compared with the control. Percent depletion of frontal cilia was most serious for the >250-<500 microm size group and least for the <63 microm size group. However, percent depletion of abfrontal cilia was most serious for the >125-<250 microm size group and least for the <63 microm size group. No recovery of ciliary damages was observed. The effects of particle size of suspended sediments on the morphological damages of gill filaments in the green-lipped mussels were discussed.

Effects of gill abrasion and experimental infection with Tenacibaculum maritimum on the respiratory physiology of Atlantic salmon Salmo salar affected by amoebic gill disease.

The effects of gill abrasion and experimental infection with Tenacibaculum maritimum were assessed in Atlantic salmon Salmo salar with underlying amoebic gill disease. The respiratory and acid-base parameters arterial oxygen tension (P(a)O2), arterial whole blood oxygen content (C(a)O2), arterial pH (pHa), haematocrit and haemoglobin concentrations were measured at intervals over a 48 h recovery period following surgical cannulation of the dorsal aorta. Mortality rates over the recovery period were variable, with gill abrasion and inoculation with T. maritimum causing the highest initial mortality rate and unabraded, uninoculated controls showing the lowest overall mortality rate. Fish with abraded gills tended to show reduced P(a)O2 and lower C(a)O2 compared with unabraded fish. Infection with T. maritimum had no effect on P(a)O2 or C(a)O2. All fish showed an initial alkalosis at 24 h post-surgery/inoculation which was more pronounced in fish inoculated with T. maritimum. There were no significant effects of gill abrasion or infection upon the ratio of oxygen specifically bound to haemoglobin or mean cellular haemoglobin concentration. Histologically, 48 h following surgery, abraded gills showed multifocal hyperplastic lesions with pronounced branchial congestion and telangiectasis, and those inoculated with T. maritimum exhibited focal areas of branchial necrosis and erosion associated with filamentous bacterial mats. All fish examined showed signs of amoebic gill disease with multifocal hyperplastic and spongious lesions with parasome-containing amoeba associated with the gill epithelium. The results suggest that respiratory compromise occurred as a consequence of gill abrasion rather than infection with T. maritimum.

Acute defense mechanisms against hemorrhage from mechanical gill injury in rainbow trout.

By cutting gill filaments in anesthetized rainbow trout (Oncorhynchus mykiss), observing the bleeding through a stereomicroscope, and using blockers of various known endogenous filament artery vasoconstrictors, we have here attempted to characterize hemostatic mechanisms in gills. The immediate hemostatic response to a cut in a gill filament artery was a local vasoconstriction, stopping the hemorrhage within approximately 20 s. In heparinized fish, the hemorrhage recommenced after approximately 8 min, suggesting that the vasoconstriction soon subsides and blood clotting becomes responsible for the hemostasis. Antagonists of acetylcholine, adenosine, and serotonin receptors were unable to block the hemostatic vasoconstriction. Also, tetrodotoxin was without effect, indicating a nonnervous origin. By contrast, indomethacin significantly affected the measured bleeding times, suggesting that eicosanoids play a significant role in this process (possibly by stimulating vasoconstriction and/or by inducing local thrombocyte aggregation). By possessing several hundred virtually identical filaments with readily observable vasculature, the fish gill appears to be a good experimental model for studying hemostatic mechanisms.

A comparative study of oxygen toxicity in vertebrates.

Survival time in conditions of hyperbaric oxygenation were measured in trout and eels in water, and in frog in water or in gas phase at various temperatures. In eel and trout, the gill surface is altered within 90 min at 15 ata pressure of oxygen. Survival times of the frog during hyperoxia in aquatic or gaseous conditions are only slightly different, in spite of the marked difference in the oxygen concentration of the two media. Oxygen toxicity is well correlated with the aerobic metabolic rate, (1) in a given species adapted to various temperatures; (2) in trout eel and frog observed at the same temperature. The differing O2 toxicities in homeothermic and poikilothermic animals are also related to the differences in metabolic activity.