Histological and hematological alterations of silver catfish Rhamdia quelen highly parasitized by Lernaea cyprinacea.

The main objective of this study was to evaluate the histological and hematological alterations of silver catfish Rhamdia quelen (Quoy & Gaimard, 1824) against intense parasitism by anchor worm Lernaea cyprinacea Linnaeus, 1758. Thirteen fish specimens were collected at the Fish Genetic Improvement Unit (EPAGRI/CEDAP), Brazil. After acclimatization, the fish were anesthetized for blood collection, and the samples were used for counting thrombocytes, total leukocytes, differential leukocytes and for quantification of total erythrocytes. Afterwards, the animals were euthanized, the parasites removed and quantified, and the portions of the affected tegument fixed in 10% buffered formalin. All the analyzed animals were parasitized by L. cyprinacea with mean intensity of 192 specimens per fish. The hematological profile of the animals was different from that reported for healthy fish. Immature leukocytes were the most abundant cells, followed by monocytes, lymphocytes, neutrophils, and periodic acid-Schiff-positive granular leukocytes. A small number of neutrophils were present in the bloodstream of parasitized fish, while histological examination evidenced the accumulation of this cell type near the place of infection by the parasite. Total leukocytes correlated significantly and positively (r = 0.6, p = 0.030) with the intensity of L. cyprinacea infection and the fish weight. Histological changes included hemorrhage, liquefactive necrosis and connective tissue disarrangement, necrosis, granulomatous inflammation, mononuclear and polynuclear inflammation, as well as hyperplasia and hypertrophy of the alarm and mucous cells. The L. cyprinacea infection intensity caused deleterious changes both in the hematological profile and the integument of R. quelen, confirming the pathogenic potential of this parasite to the affected fish species.

Trace elements and microbiological parameters in farmed Nile tilapia with emphasis on muscle, water, sediment and feed.

Fish may bioaccumulate contaminants from the aquatic environment and extend them to the food chain provoking risks to human health. This study evaluated the microbiological parameters of the pond´s water and trace elements concentrations in samples of water, sediment, feed and muscle of farmed Nile tilapia used for human consumption in southern Brazil. A total of 240 fish were collected from 12 tilapia farms. Sediment, tank water and dry ration used in the animals' diet were collected for analysis. Analysis were performed by Energy Dispersion X-ray Fluorescence (EDXRF), Inductively Coupled Plasma Mass Spectrometer (ICP-MS), Induced Coupled Plasma Optical Emission Spectrometer (ICP-OES), and Atomic Absorption Spectrometer (AAS-VGA). In addition, the microbiological analysis of the water was carried out. The concentrations of Se, I, Fe, Cu and Zn in fish muscle were higher than the recommended by the Brazilian legislation, considering the advised daily intake for adults. The arsenic element had concentrations above the limit stipulated by the present Brazilian legislation, observed in all samples of muscle, sediment and tank water highlighting a possible environmental and fish contamination by the toxic element. Moreover, the arsenic concentration in the water presented a positive correlation (ρ = 0.33) with arsenic in the muscle, suggesting that tilapia is a good environmental bioindicator, once they properly reflect the levels of arsenic in the water. It is suggested to perform an arsenic speciation for quantification of the inorganic form and accurate assessment of the degree of toxicity in the muscle samples and risks it can bring to human health. Regarding the other potentially toxic elements (Hg, Pb and Cd), and microbiological analysis of water it was verified that the consumption of the fish in question does not raise risks, since the values are within a quality benchmark established by law. The concentration of total and fecal coliforms in pond´s water in the facilities was in agreement with the microbiological indexes required by the legislation of CONAMA class II. Western region presented the lowest concentrations of fecal coliforms when compared to the other regions. There was no significant difference in the microbiological counts of total heterotrophic bacteria, Vibrio spp. and Pseudomonas spp. among the regions.

Lippia sidoides essential oil at concentration of 0.25% provided improvements in microbiota and intestine integrity of Danio rerio.

The study evaluated the effects of dietary supplementation with Lippia sidoides essential oil on the microbiota and intestinal morphology of Danio rerio. For this, 448 fish were randomly distributed in 28 tanks divided into a control group fed a commercial diet without supplementation, a group fed a commercial diet containing grain alcohol and five groups fed a commercial diet containing essential oil of L. sidoides (LSEO) at concentrations of 0.25%, 0.50%, 0.75%, 1.00% and 1.25%. After the period of dietary supplementation, biological materials were collected for microbiological and histological analyses. There were no significant differences regarding the microbiological count between the groups. Diversity of the microbiome was higher in 0.25% group than in control group. LSEO inhibited the growth of potentially pathogenic bacteria. Fish fed LSEO0.25% showed greater intestinal histomorphometric indices. The inclusion of LSEO at 0.25% in the diet of D. rerio provided improvements in fish microbiota and intestine integrity.

O estudo avaliou os efeitos da suplementação dietária com óleo essencial de Lippia sidoides sobre a microbiota e a morfologia intestinal de Danio rerio. Para isso, 448 peixes foram distribuídos aleatoriamente em 28 tanques divididos em grupo controle alimentado com dieta comercial sem suplementação, grupo alimentado com dieta comercial contendo álcool de cereais e cinco grupos alimentados com dieta comercial contendo óleo essencial de L. sidoides (LSEO) em concentrações de 0,25%, 0,50%, 0,75%, 1,00% e 1,25%. Após o período de suplementação alimentar, foram coletadas amostras biológicas para análises microbiológicas e histológicas. Não houve diferenças significativas quanto à contagem microbiológica entre os grupos. A diversidade do microbioma foi maior no grupo de 0,25% do que no grupo controle. LSEO inibiu o crescimento de bactérias potencialmente patogênicas. Peixes alimentados com LSEO 0,25% apresentaram maiores índices histomorfométricos intestinais. A inclusão de LSEO a 0,25% na dieta de D. rerio proporcionou melhorias na microbiota e na integridade intestinal dos peixes.

Influence of Seasonality and Culture Stage of Farmed Nile Tilapia (Oreochromis niloticus) with Monogenean Parasitic Infection.

The aim of this study was to observe how abiotic and biotic factors in a tropical region influence the rate of monogenean parasitism in Nile tilapia (Oreochromis niloticus) that are farmed in net cages. A total of 240 sexually reversed fish were analyzed, and 20 from each culture stage were collected during each sampling month. Overall, 60 fish were sampled in April (autumn), 60 in August (winter), 60 in November (spring), and 60 in February (summer). Fish were collected from a commercial fish farm located in Capivara Reservoir in the lower Paranapanema River region of Paraná, Brazil. In total, 3290 monogenean parasites were collected from fish gills of the following species: Cichlidogyrushalli, C. thurstonae, Scutogyruslongicornis, C. cirratus, C. sclerosus, and C. tilapiae. Higher parasitological indices were observed in colder seasons with lower precipitation. Autumn had the highest parasitic infection values compared to the other seasons. The occurrence of monogenean parasites showed a negative correlation with season, in contrast to the culture stage, in which there was a positive correlation. These results may provide a means for establishing adequate fish farm management to predict periods of high monogenean infestation.

Neobenedenia melleni from reef ornamental fish species in a retailer of Southeastern Brazil and its possible role as a mechanical vector of bacterial infection.

Annually, more than 2500 ornamental fish species are traded worldwide. Forty percent of these are from marine water. Some 98% of marine species are wild-caught from their natural habitat, and the majority subsequently exported. Wild fish frequently carry pathogens, which could induce diseases after the stress of capture. Neobenedenia melleni is a platyhelminth that mainly attaches to the skin and eyes of the host. It provokes dermal inflammation, epidermal loss, skin depigmentation, reduction in the number of mucous cells, and, consequently, decreased mucus protection, and declining immunological barriers. This makes fish susceptible to secondary infections. A total of 47 wild reef fish from a retailer were examined, suspected to be infected with ectoparasites. The morphological identification revealed N. melleni as a monogenean agent. One monogenean specimen was collected from the eye of each of the 40 fish analyzed to evaluate possible bacterial secondary infections using the matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) technique. The MALDI-TOF MS identified that 59% of monogenean collected from the eyes had bacteria, including some pathogenic to fish. This led us to believe that the ectoparasite can be a possible mechanical vector of pathogenic bacteria for fish culture and maintenance. The use of praziquantel as an antiparasitic agent is also discussed.

Antiparasitic potential of alternative treatments against larval stages of Lernaea cyprinacea.

This study evaluated the potential of alternative treatments against larval stages of Lernaea cyprinacea. For in vitro test, the nanoemulsified oils of Pinus sp. acicule and oleoresin were evaluated, along with Biogermex® (commercial product based on citrus biomass). For this, the motility of five larvae of the same stage (nauplii or copepodite) were evaluated in a 96-well microplate. Using the best results, on the in vivo test, fries of Rhamdia quelen were submitted to a long-term immersion bath (96 h) containing different concentrations of the product diluted directly in the water. It was possible to notice the antiparasitic potential of the oleoresin and the acicule of Pinus sp., as well as the citrus biomass extract against the parasites. The nanoemulsified oils successfully inhibited the development of nauplii (10 mg L-1 in 24 h) and the fries showed to be tolerant to the presence of the compound (LC50 96 h-16.74 mg L-1). The concentration of 30.5 mg L-1 of Biogermex® eliminated the copepodites within 24 h, being more efficient than Pinus sp. when tested at the same stage, at the times analyzed. The results obtained indicate a potential use of these compounds as prophylactic agents against L. cyprinacea.

Emamectin Benzoate Treatment of Hybrid Grouper Infected With Sea Lice in Hong Kong.

Sea lice (Copepoda: Caligidae) are ectoparasites which negatively impact marine aquaculture species around the world. There are a limited number of treatments licensed for use against sea lice in tropical and semi-tropical farmed fish species. Emamectin benzoate (EB) was an effective pharmaceutical drug against sea lice infestations in several salmon industries before resistance to the product developed. This drug has not been extensively tested in marine fish within Asia. The objective of this study was to determine whether this drug could be used to treat oral infections with sea lice in hybrid grouper (Mycteroperca tigris × Epinephelus lanceolatus) cultured in saltwater net-pen sites in Hong Kong. We observed an overall reduction in sea lice infections over time, starting on the last day of the treatment up to the end of our study (i.e., 14 days after the last EB treatment). We also observed a large variation in concentrations of EB in fish on the last day of the treatment, which provides an explanation for the variation in response to the treatment. It also suggests that distribution of the medication to fish in saltwater net-pens is difficult, especially when medication is hand-mixed in the feed and possibly unevenly distributed in the daily rations. Overall, this study provides preliminary evidence that EB could be used to treat sea lice found in Hong Kong and potentially in other regions of SE Asia.

Haliotrema aurigae (Yamaguti, 1968) Plaisance, Bouamer & Morand, 2004, in ornamental reef fish imported into Brazil.

More than 1.450 marine fish species are commercialized as ornamental fish and approximately 90% of these are wild-caught. Rainford's butterflyfish, Chaetodon rainfordi (McCulloch, 1923), is endemic to the western Pacific. This fish species belongs to one of the main ornamental fish families imported into Brazil. Haliotrema is a genus of the family Dactylogyridae. Species of this genus have very interesting host associations and geographic distributions. The aim of this study was to report on occurrences of exotic Haliotrema aurigae in diseased Rainford butterflyfish that were imported to Brazil. One Rainford's butterflyfish with suspected parasitic disease was euthanized and subjected to skin and gill scraping. The parasite specimens were mounted on semi-permanent slides with Hoyer's solution for morphological analysis of sclerotized structures. The hard-sclerotized structures of these specimens were compared and were identified as those of Haliotrema aurigae (Yamaguti, 1968). These results emphasize the importance of implementation of correct control measures at the country's borders, to avoid the introduction of exotic parasites. In addition, it can be emphasized that the easy adaptation of this parasite to new environments is a concern for ornamental fish farmers.

Parasitological and histopathological diagnosis of a non-native fish (Oreochromis sp.) with a noticeable presence in a natural Brazilian river environment.

Nile tilapia is an exotic species produced in Brazilian aquaculture. The aim of this study was to constitute a pathological diagnosis in Oreochromis sp. with a noticeable presence in a river in southern Brazil, and to verify the origin of these fish in this natural habitat, an environment with high sanitary precariousness due to discharges of industrial effluents. A total of 30 fish were collected during three sampling events and their health status was assessed by parasitological fauna survey, parasitological indices and histopathological examination of the gills, liver, spleen and kidney. Five species of monogenea were identified infesting gills and stomach: Cichlidogyrus sclerosus, C. halli, C. thurstonae, Scutogyrus longicornis and Enterogyrus cichlidarum; and 4 species of trichodinidae infesting the body surface and gills: Trichodina magna, T. compacta, T. centrostrigeata and Paratrichodina africana. The results showed differences according to the seasonality of the collection, with higher parasite count and larger tissue alterations in summer. Histopathological analysis fluctuated from moderate to severe, but necrosis prevailed in all organs and in all fish, indicating that these fish did not enjoy full health.