Mitochondrial genome of Rhabdosynochus viridisi (Monogenea: Diplectanidae), a parasite of Pacific white snook Centropomus viridis.

We report the nearly complete mitochondrial genome of Rhabdosynochus viridisi - the first for this genus - achieved by combining shotgun sequencing of genomic and cDNA libraries prepared using low-input protocols. This integration of genomic information leads us to correct the annotation of the gene features. The mitochondrial genome consists of 13,863 bp. Annotation resulted in the identification of 12 protein-encoding genes, 22 tRNA genes and two rRNA genes. Three non-coding regions, delimited by three tRNAs, were found between the genes nad5 and cox3. A phylogenetic analysis grouped R. viridisi with three other species of diplectanid monogeneans for which mitochondrial genomes are available.

Silver nanoparticles are lethal to the ciliate model Tetrahymena and safe to the pike silverside Chirostoma estor.

Ciliate ectoparasites are one of the most important groups of pathogens in fish culture, and the traditional treatments are sometimes harmful to the fish and the environment. Thus, the search for novel compounds that are effective at low concentrations and safe for fish are necessary to optimise treatments in aquaculture. The antiprotozoal capacity of silver nanoparticles (AgNPs) against the ciliate Tetrahymena has been documented; however, their toxicity may vary with the synthesis methodology and nanoparticle size. The objectives of this study were a) to evaluate the acute toxicity in vitro of two AgNPs (Argovit™ and UTSA) on Tetrahymena sp., a biological model for ciliated ectoparasites of fish and b) to test the safety of lethal and higher doses of UTSA AgNPs for ciliates on the fish C. estor. Light microscopy and scanning electron microscopy (SEM) were used to determine whether AgNPs affected the structure of the cell surface of Tetrahymena. The mortality, histopathological alterations and metagenomics of the fish were used to determine the major effects of UTSA AgNPs. In Tetrahymena, the median lethal concentration (LC50) for Argovit™ was 2501 ± 1717 ng/L at 15 min and 796 ± 510 ng/L at 60 min, while the LC50 for UTSA AgNPs was 4 ± 2 and 1 ± 0.6 ng/L at 15 min and 60 min, respectively. A concentration of 3300 ng/L Argovit™ and 10.6 ng/L UTSA AgNPs for 15 and 60 min, respectively, was 100% effective against Tetrahymena. After 60 min of exposure to 0.25 and 0.50 ng/L UTSA AgNPs, the number of cilia significantly reduced, there were small holes on the cell surface, and the cellular membrane was ruptured. In fish exposed to lethal (10.6 ng/L) and higher (31.8 and 95.4 ng/L) doses of UTSA, the AgNPs did not affect fish survival after 96 h, and there were no signs of histopathological damage or gut microbial changes. This study is the first report on microscopic and ultrastructural changes in Tetrahymena after exposure to significantly low concentrations of UTSA AgNPs with antiprotozoal efficacy without evidence of harmful effects on fish. These results provide the basis for further studies of both pet aquarium and commercial fish that may validate these findings at a larger experimental scale, taking into account AgNPs bioaccumulation, safety for human consumption and environmental impact.

Quantitative real-time PCR (qPCR) for the detection and quantification of dactylogyrid parasites infecting Lutjanus guttatus.

Severe infections of the spotted rose snapper Lutjanus guttatus resulting from dactylogyrid monogeneans present a risk to aquaculture. Currently, the diagnosis of this infection requires the morphological identification and manual quantification of parasites. Based on the characterization of the 28S rRNA gene of dactylogyrid species present in L. guttatus, specific primers were designed for real-time polymerase chain reaction (qPCR) using EvaGreen® chemistry. The standard curve method estimated the number of dactylogyrids accurately. A total of 85 gill samples from cage-cultured fish infected with dactylogyrids were analysed. The estimated number of dactylogyrids using this molecular method was very similar to the manual count that was performed initially. The standardized qPCR approach will be helpful as a complementary method for the early routine monitoring of dactylogyrid infections and for epidemiological studies in which a high number of fish must be studied.

Influence of ß-glucan on innate immunity and resistance of Lutjanus guttatus to an experimental infection of dactylogyrid monogeneans.

The present study investigated the effect of ß-1,3/1,6-glucan on growth, haematology, innate immunity and resistance against dactylogyrids on the spotted rose snapper (Lutjanus guttatus). Fish were fed during 5 weeks with commercial diet (control group) and same diet supplemented with three levels of ß-glucans (0·05%, 0·1% and 0·5%/kg feed). The results showed that at concentrations of 0·05% and 0·1%, fish growth was enhanced, and in weeks 2 and 4, an increase in cellular responses such as percentage of monocytes, neutrophils, respiratory burst activity and nitric oxide activity was observed. In diet with 0·5%ß-glucans, changes were registered at the end of the experiment. At week 2, 0·05%ß-glucans showed the best response to most of the analysed parameters. In a second trial, diet with 0·05%ß-glucans was chosen to prove its effect on the resistance of infected fish with monogeneans. The results showed that fish reduce significantly the number of dactylogyrids, and parameters such as WBC, percentage of lymphocytes, neutrophils and eosinophils were increased. In addition, WBC and percentage of thrombocytes increased in the control (+). The ß-glucans have the potential to be use in diet formulations of the spotted rose snapper and to limit the adverse effects of dactylogyrids.

Hematological characteristics of the spotted rose snapper Lutjanus guttatus (Steindachner, 1869) healthy and naturally infected by dactylogyrid monogeneans.

The aim of the present study was to obtain a basic knowledge of the hematology in order to determine changes in blood parameters of the spotted rose snapper Lutjanus guttatus. The morphological features of blood cells were described according to the observations made by light microscopy of Wright-Giemsa-stained blood films. The reference intervals and the mean value were determined for each hematological parameter evaluated in healthy fish and data were compared to those of naturally infected, with dactylogyrid monogeneans fish. Infected fish showed a prevalence of 100% and a mean intensity of 246.6 parasites per fish. Mean values of HCT, WBC, thrombocytes percentage and eosinophils percentage were significantly higher (P<0.05) in the infected fish. In addition, lymphocytes percentage and total protein were significantly lower (P<0.05) in the infected fish compared to healthy fish. Only total WBC count, lymphocytes percentage and eosinophils percentage in infected fish were outside reference interval. The hematology of the spotted rose snapper of this study might serve as a basis for future studies and diagnosis. Changes observed in blood parameters in infected fish suggest that the immune system of L. guttatus was affected by the presence of the parasites.

Philometra floridensis sp. n. (Nematoda: Philometridae) from the ovary of red drum Sciaenops ocellatus (Osteichthyes: Sciaenidae) off the coast of Florida, USA.

A new nematode species, Philometra floridensis sp. n. (Philometridae), is described from male and female specimens found in the ovary of red drum, Sciaenops ocellatus (Linnaeus) (Sciaenidae, Perciformes), from the Gulf of Mexico off Treasure Island, Florida, USA. Based on light and scanning electron microscopy examination, the new species differs from most other gonad-infecting Philometra spp. in having a smooth gubernaculum with a distinct dorsal tooth on the distal tip. The new species is most similar to P. carolinensis Moravec, de Buron & Roumillat, 2006, but differs in length and shape of spicules. It can be distinguished from P. carolinensis and other species with unknown males, by the markedly greater body length of gravid females (up to about 100 cm). Philometra floridensis is the third valid gonad-infecting species of Philometra reported from sciaenids.

Copepods and larvae of nematodes parasitizing (correction of parasiting) the white mullet Mugil curema (Valenciennes, 1836): indicators of anthropogenic impacts in tropical coastal lagoons?

The relationship between parasites and environmental stress were studied in two tropical coastal lagoons of Northwest Mexico: Urias estuary (highly polluted) and Teacapan estuary (slightly polluted). Metazoan parasites were examined in 292 white mullet (Mugil curema) specimens collected bimonthly during a year from both systems. Haliotrema mugilinus, Metamicrocotyla macracantha, Ergasilus sp., Caligus sp., Holobomolochus sp., and Lernaeopodidae were found in gills, while Contracaecum sp. larvae III was found liver, hepatic portal vein and kidneys. Ecological indices were influenced by the slightly higher number of parasitic species in Urias compared to Teacapan, as well as the clear dominance of two species: Ergasilus sp. and Contracaecum sp. in both systems. In fact, Ergasilus sp. showed considerably higher abundance in Urias, possibly indicating that its success was a result of adverse conditions affecting the host, while Contracaecum sp showed higher abundances in Teacapan, suggesting that the environmental conditions occurring in Urias could have produced negative impacts on the nematode's infective potential.