Hemiurid and lecithasterid digenean trematodes and camallanid and cucullanid nematodes parasitizing flounders collected off the coast of Rio de Janeiro State, Brazil / Trematodeos digeneticos hemiurideos e lecitasterideos e nematoides camalanideos e cuculanideos parasitando linguados coletados na costa do Estado do Rio de Janeiro, Brasil

Abstract A total of 132 flounder specimens (60 Paralichthys isosceles, 36 Paralichthys patagonicus and 36 Xystreurys rasile) were collected off the coast of the state of Rio de Janeiro, Brazil. The fish were measured, necropsied, and had their organs investigated for hemiurid and lecithasterid digenean trematodes and camallanid and cucullanid nematodes. Taxonomic identification of the parasites was based on morphological and morphometric characters and was conducted using bright-field and scanning electron microscopies. The trematodes Lecithochirium monticellii and Aponurus laguncula were found parasitizing P. isosceles, P. patagonicus and X. rasile while the nematodes Procamallanus (Spirocamallanus) halitrophus and Cucullanus bonaerensis were found parasitizing P. isosceles and X. rasile and P. isosceles, P. patagonicus and X. rasile, respectively. Parasite indices of prevalence, mean intensity, mean abundance, and range of infection, as well as infection site, were evaluated for each parasite species. This study allowed to evidence the first occurrence of P. patagonicus by L. monticellii; X. rasile by A. laguncula and P. (S.) halitrophus; and P. isosceles and P. patagonicus by C. bonaerensis in the Western South Atlantic Ocean.

Resumo Um total de 132 espécimes de linguados (60 Paralichthys isosceles, 36 Paralichthys patagonicus e 36 Xystreurys rasile) foram coletados na costa do Estado do Rio de Janeiro, Brasil. Os peixes foram medidos, necropsiados e tiveram seus órgãos investigados para o encontro de trematodeos digenéticos, das famílias Hemiuridae e Lecithasteridae, e nematoides das famílias Camallanidae e Cucullanidae. A identificação taxonômica dos parasitos foi baseada nos caracteres morfológicos e morfométricos, usando-se as microscopias de campo claro e eletrônica de varredura. Os trematódeos, Lecithochirium monticellii e Aponurus laguncula, foram encontrados parasitando P. isosceles, P. patagonicus e X. rasile, enquanto os nematoides, Procamallanus (Spirocamallanus) halitrophus e Cucullanus bonaerensis, foram encontrados parasitando P. isosceles e X. rasile e P. isosceles, P. patagonicus e X. rasile, respectivamente. Os índices parasitários de prevalência, intensidade média, abundância média e amplitude de variação da intensidade de infeção, bem como os sítios de infecção foram avaliados para cada espécie de parasito. Este é o primeiro registro de P. patagonicus por L. monticellii; X. rasile por A. laguncula e P. (S.) halitrophus; e de P. isosceles e P. patagonicus por C. bonaerensis no Oceano Atlântico Sul Ocidental.

Flatfish (Pleuronectiformes) species richness and depth distribution in the Gulf of Nicoya, Golfo Dulce, and two adjacent areas on the Pacific coast of Costa Rica / Riqueza de especies de lenguados (Pleuronectiformes) y distribución por profundidad en el Golfo de Nicoya, Golfo Dulce y dos áreas adyacentes en la costa del Pacifico de Costa Rica

Introduction: Information on the number of flatfish species and their depth distributions is scarce for the Eastern Tropical Pacific. Historical data is important to evaluate changes in ecosystems due to local, regional and global stressors. Objective: To provide information on the presence, depth distributions and lengths of flatfish species collected by trawl nets. Methods: Fish studies were conducted by trawling at four sites along the Pacific coast of Costa Rica by the survey vessels Skimmer (1979-1980), Nishin Maru (1987-1988) and Victor Hensen (1993-1994). The published lists of species were consulted, museum specimens were accessed, and an updated list of flatfish species assembled. Results: A total of 36 species were found over a depth range of 8-359 m. The family Paralichthyidae included 13 species followed by the Cynoglossidae with 12 species, Achiridae with six and Bothidae with five. Thirty-five species were collected at depths from 10-235 m in the Gulf of Nicoya and Golfo Dulce. Monolene asadeae was collected offshore at a depth range of 254-359 m. The Gulf of Nicoya estuary yielded 27 species during the Skimmer expedition over a depth range of 8-60 m, while 31 species were found during the V. Hensen survey at depths from 10-228 m. The V. Hensen survey in Golfo Dulce (20-235 m) collected 19 species, and 17 species in Coronado Bay (21-187 m). Off the Nicoya Peninsula (56-359 m) 13 species were collected by the Nishin Maru shrimp trawler. A total of 21 species (58 %) were found at depths greater than 100 m which were probably exposed to low oxygen concentrations. During the V. Hensen survey Symphurus chabanaudi and S. elongatus were collected more frequently in the Gulf of Nicoya, while S. leei was collected more frequently in Golfo Dulce. Cluster analyses based on presence-absence data for the Gulf of Nicoya and Golfo Dulce revealed low station similarity indicating possible habitat partitioning among species. The size (Total length) of 33 species measured from both the Gulf of Nicoya and Golfo Dulce ranged from 8 cm (Syacium cf longidorsale, Trinectes xanthurus) to 50 cm (S. ovale). Only 12 species were found with lengths over 20 cm. Data on flatfish landings by the semi-industrial fishing fleet for the period 2000-2016 indicates that this group represents less than 1 % of the total landings (shrimp and bycatch), with a minimum of 365 kg in 2001 and a maximum of 13 414 kg in 2013. Conclusions: The number of flatfish species of the Pacific coast of Costa Rica appears relatively high but comparable to the numbers found in other tropical regions. A reduced fishing impact on the populations together with this updated list of flatfish provide a good baseline for a new survey of fish populations. Both are important for a future updating of the trophic models available the Gulf of Nicoya and Golfo and their use as tools for better management of the ecosystems.

Introducción: Es escasa la información sobre el número de especies y la distribución batimétrica de los lenguados del Pacífico Este Tropical. Esos datos son importantes para evaluar cambios en los ecosistemas debidos a tensores locales, regionales y globales. Objetivo: Proveer información de la presencia, distribución batimétrica y longitudes de especies de lenguados recolectados por red de arrastre. Métodos: Evaluaciones de los peces mediante redes de arrastre fueron conducidas en cuatro sitios a lo largo de la costa del Pacífico de Costa Rica por los buques de investigación Skimmer (1979-1980), Nishin Maru (1987-1988) y Victor Hensen (1993-1994).Se consultó las listas publicadas de especies, se revisó ejemplares depositados en el museo y se integró una lista actualizada de especies de lenguados. Resultados: Un total de 36 especies de lenguados fueron encontrados en un ámbito de profundidad de 8 a 359 m. La familia Paralichthyidae incluyó 13 especies seguida por los Cynoglossidae con 12 especies, Achiridae tuvo seis y Bothidae cinco especies. Treinta y cinco especies fueron recolectadas entre 10 y 235 m en el Golfo de Nicoya y Golfo Dulce. Monolene asadeae fue recolectada aguas afuera en un ámbito de profundidad de 254-359 m. El Golfo de Nicoya produjo 27 especies durante la expedición del Skimmer y en un ámbito de profundidad de 8 a 60 m, mientras que 31 especies fueron encontradas durante el muestreo del V. Hensen en profundidades entre los 10 y 228 m. El muestreo del V. Hensen en el Golfo Dulce (20 a 235 m) produjo 19 especies y 17 en Bahía de Coronado (21-187 m). Afuera de la península de Nicoya (56-359 m) 13 especies fueron recolectadas por el camaronero Nishin Maru. Un total de 21 especies (58 %) fueron encontradas a profundidades mayores de 100 m y posiblemente expuestas a las bajas concentraciones de oxígeno. Durante el muestreo del V. Hensen, Symphurus chabanaudi y S. elongatus fueron capturados más frecuentemente en el Golfo de Nicoya, mientras S. leei lo fue en Golfo Dulce. El tamaño (Longitud Total) de 33 especies medidas en ambos Golfo de Nicoya y Golfo Dulce oscilaron entre 8 cm (Syacium cf longidorsale, Trinectes xanthurus) y 50 cm (S. ovale). Solo 12 especies fueron encontradas con longitudes mayores de 20 cm. Datos de desembarcos de lenguados por la flota semi-industrial para el periodo 2000-2016 indican que este grupo representa menos del 1 % de los desembarcos totales (camarones y fauna acompañante), con un mínimo de 365 kg en 2001 y un máximo de 13 414 kg en 2013. Conclusiones: Los análisis de conglomerados basados en datos de presencia-ausencia para el Golfo de Nicoya y Golfo Dulce revelaron baja similitud de estaciones indicando partición del hábitat entre las especies. La diversidad de la fauna de lenguados de la costa del Pacífico de Costa Rica aparece relativamente alta pero comparable con la encontrada en otras regiones tropicales. El número de buques arrastreros ha disminuido significativamente en el Pacífico de Costa Rica después del 2014. Esta reducción del impacto en las poblaciones y la lista actualizada de lenguados proveen una buena base para un nuevo estudio de las poblaciones de peces. Ambas son importantes para una futura actualización de los modelos tróficos disponibles para el Golfo de Nicoya y Golfo Dulce y su utilidad como herramientas para un mejor manejo de los ecosistemas.

Acanthocephalan parasites of the flounder species Paralichthys isosceles, Paralichthys patagonicus and Xystreurys rasile from Brazil / Acantocéfalos parasitos de espécies de linguados Paralichthys isosceles, Paralichthys patagonicus e Xystreurys rasile no Brasil

Abstract Flounders are commercially and economically important fish. A total of 120 specimens of flounders (60 Paralichthys isosceles, 30 Paralichthys patagonicus and 30 Xystreurys rasile) were collected off the coast of the state of Rio de Janeiro, Brazil. The fish were measured, necropsied and filleted, and then had their organs investigated for acanthocephalans. Taxonomic identification of the parasites was based on morphological, morphometric and genetic characters. Paralichthys isosceles and P. patagonicus were parasitized by juveniles of Serrasentis sagittifer, Bolbosoma turbinella, Corynosoma australe and C. cetaceum; Xystreurys rasile was parasitized by C. australe. Genetic characterization confirmed the identification of specimens of Bolbosoma turbinella and Corynosoma australe, as demonstrated by phylogenetic analyses using both ITS and cox1 molecular targets. Parasite indices of prevalence, intensity, mean intensity, abundance, mean abundance, and range of infection, as well as infection site, were evaluated for each parasite species. This is the first report of S. sagittifer parasitizing P. isosceles and P. patagonicus, and B. turbinella parasitizing P. patagonicus.

Resumo Os linguados são peixes comercial e economicamente importantes. Um total de 120 espécimes de linguados (60 Paralichthys isosceles, 30 P. patagonicus e 30 Xystreurys rasile) foram coletados no litoral do estado do Rio de Janeiro, Brasil. Os peixes foram medidos, necropsiados, filetados e tiveram seus órgãos investigados para a presença de acantocéfalos. A identificação taxonômica foi baseada em caracteres morfológicos, morfométricos e genéticos. Paralichthys isosceles e P. patagonicus estavam parasitados por acantocéfalos juvenis de Serrasentis sagittifer, Bolbosoma turbinella, Corynosoma australe e C. cetaceum; Xystreurys rasile estava parasitado com C. australe. A caracterização genética confirmou a identificação dos espécimes de Bolbosoma turbinella e Corynosoma australe, como demonstrado por análises filogenéticas usando ambos marcadores moleculares ITS e cox1. Foram analisados os índices parasitários: prevalência, intensidade, intensidade média, abundância, abundância média, amplitude de variação da infecção e sítio de infecção de cada espécie de parasito. Este é o primeiro registro de S. sagittifer parasitando P. isosceles e P. patagonicus, e de B. turbinella parasitando P. patagonicus.

Pterobothrium sp. parasitando linguado, Xystreurys rasile (Jordan, 1891), coletados no litoral do Estado do Rio de Janeiro, Brasil e sua importância na inspeção do pescado / Pterobothrium sp. parasitizing flounders, Xystreurys rasile (Jordan, 1891), collected off the coast the State of Rio de Janeiro, Brazil and its fishing inspection importance

Alguns parasitos, podem conferir aspecto repugnante ao pescado, sendo condenados pelo Serviço de Inspeção, quando na indústria de beneficiamento. Entre novembro de 2010 e maio de 2018 foram adquiridos 36 espécimes de Xystreurys rasile em mercados de pescados no município do Rio de Janeiro, Brasil. Os peixes foram identificados, medidos, pesados e necropsiados. Os cestoides foram processados segundo as técnicas usuais em helmintologia. Cinco linguados estavam parasitados por 29 cestoides do gênero Pterobothrium, os índices parasitários foram: prevalência (14%), intensidade média (5,8), abundância média (0,8) e amplitude de infecção (2-12) e os sítios de infecção foram estômago, intestino, rim, fígado, serosas do estômago, baço e fígado e cavidade abdominal. Este é o primeiro registro de Pterobothrium sp. parasitando X. rasile.

Kudoa ogawai (Myxosporea: Kudoidae) Infection in Cultured Olive Flounder Paralichthys olivaceus

Since Kudoa septempuntata was identified as a causative agent of food poisoning associated with raw olive flounder Paralichthys olivaceus, interest and concern regarding the parasite have increased. However, there have been no investigations or reports of other Kudoa species infecting the fish (except for K. paralichthys, which infects the brain) in Korea. We found cysts filled with myxospores of Kudoa species in muscles of cultured olive flounder specimens and identified these to the species level. Mature spores were quadrate, measuring 8.7±0.5 μm in length, 9.2±0.4 μm in thickness, and 12.9±0.6 μm in width. The spores containing 4 polar capsules had a length of 2.1±0.2 μm and a width of 1.8±0.3 μm. The partial 18S and 28S rDNA of isolates showed 99–100% similarities with K. ogawai. Using these morphological and molecular analyses, the species was identified as K. ogawai. This study is the first report of K. ogawai infection in cultured olive flounder in Korea.

Sinuolinea capsularis (Myxosporea: Sinuolineidae) Isolated from Urinary Bladder of Cultured Olive Flounder Paralichthys olivaceus

Sinuolinea capsularis Davis, 1917 is myxosporean that infect the urinary system of the host fish. Insufficient morphological and molecular data of S. capsularis exits, and it is therefore difficult to make an accurate identification of the parasite. We tried a series of morphological and molecular analysis to identify an myxosporean isolated from urinary bladder of cultured olive flounder, Paralichthys olivaceus, from Jeju island in the Republic of Korea. Some of them were observed under a light microscope and SEM, and remain samples were used molecular and phylogenetic analysis. Mature spores were subspherical, measuring 13.9±0.6 μm in length and 13.8±0.8 μm in width. Two spherical polar capsules on opposite sides in the middle of the spore had a diameter range of 4.3±0.4 μm. Scanning electron microscopy revealed that spores a severely twisted the suture line. By the morphological comparison and analysis, it was identified as S. capsularis. In addition, we obtained the partial 18S rDNA of S. capsularis and first registered it in NCBI. Phylogenetic analysis showed that S. capsularis clustered with Zschokkella subclade infecting the urinary system of marine fish, and it supported the infection site tropism effect on phylogeny of marine myxosporeans as well as the origin of Sinuolinea is not monophyly.

Characterization of digestive enzymes from captive Brazilian flounder Paralichthys orbignyanus / Caracterização de enzimas digestivas do linguado brasileiro de cativeiro Paralichthys orbignyanus

Abstract Knowledge of specific enzyme activity, along with animal habits and digestive capacity is essential in formulating an appropriate diet for any species. In this study, we evaluated and characterized the activity of digestive enzymes present in the liver, intestine, and stomach of Paralichthys orbignyanus. The effects of pH and temperature on enzyme activity were also evaluated via the use of specific substrates. The use of specific substrates and inhibitors showed strong evidence of the presence of trypsin (BApNA= 0.51 ± 0.2 mU mg-1), chimotrypsin (SApNA= 2.62 ± 1.8 mU mg-1), and aminopeptidases (Leu-p-Nan =0.9709 ± 0.83 mU mg-1) in the intestine. Optimum pH for the activity of trypsin, chemotrypsin, leucino aminopeptidase, amilase, and pepsin were 9.5, 9.0, 8.0, 7.5, and 3.5, respectively, while optimum temperatures were 50, 50, 50, 40, and 45 °C, respectively. These results provide additional information regarding the biology of Brazilian flounder and can be used as a basis for further studies regarding fish feeding physiology.

Resumo O conhecimento da atividade enzimática é essencial para formular uma correta dieta específica para espécie, além de estarem correlacionadas com o hábito da alimentação e capacidade digestive. Neste estudo determinamos e caracterizamos a atividade enzimática presente no intestino, estômago e fígado do linguado Paralichthys orbignyanus. Os efeitos da temperatura e pH sobre a atividade enzimática também foram avaliados utilizando substratos específicos. O uso de substratos e inibidores específicos mostrou uma forte evidência da presença da tripsina (BApNA = 0,51 ± 0,2 mU mg-1), quimotripsina (SAPNA = 2,62 ± 1,8 mU mg-1), e as aminopeptidases (Leu-p-Nan = 0,97 ± 0,83 mU mg-1) no intestino. O pH ótimo observado para a atividade de tripsina, quimotripsina, leucino aminopeptidase, amilase e pepsina foi 9,5, 9,0, 8,0, 7,5 e 3,5, respectivamente. A temperatura ótima observada foi 50, 50, 50, 40 e 45 °C, respectivamente. Estes resultados fornecem informações adicionais sobre a biologia do linguado brasileiro e pode ser usado como base para novos estudos sobre fisiologia alimentar.

Foodborne Illness Outbreaks in Gyeonggi Province, Korea, Following Seafood Consumption Potentially Caused by Kudoa septempunctata between 2015 and 2016

OBJECTIVES: Investigations into foodborne illness, potentially caused by Kudoa septempunctata, has been ongoing in Korea since 2015. However, epidemiological analysis reporting and positive K septempunctata detection in feces in Korea has been limited. The aim of this study was to provide epidemiologic data analysis of possible food poisoning caused by K septempunctata in Korea. METHODS: This study reviewed 16 Kudoa outbreak investigation reports, including suspected cases between 2015 and 2016 in Gyeonggi province, Korea. Suspected Kudoa foodborne illness outbreak was defined as “evidence of K septempunctata in at least one sample.” The time and place of outbreak, patient symptoms and Kudoa (+) detection rate in feces was analyzed. RESULTS: Kudoa foodborne illness outbreaks occurred in most patients in August (22.6%) and in most outbreaks in April (25%). The attack rate was 53.9% and the average attack rate in patients who had consumed olive flounder was 64.7%. The average incubation period was 4.3 hours per outbreak. Diarrhea was the most common symptom which was reported by 91.5% patients. The Kudoa (+) detection rate in feces was 69.2% of cases. CONCLUSION: Monthly distribution of Kudoa foodborne illness was different from previous studies. The Kudoa (+) detection rate in feces decreased rapidly between 25.5 and 28.5 hours of the time interval from food intake to epidemiologic survey. To identify effective period of time of investigation, we believe additional study with extended number of cases is necessary.

Analysis of Kudoa septempunctata as a cause of foodborne illness and its associated differential diagnosis / 한국역학회지

OBJECTIVES: Recently kudoa septempuctata in olive flounders is suggested as a cause of food poisoning, however whether kudoa septempuctata can affect human gastrointestinal systems is controversial and its pathogenecity remains unclear. In view of the field epidemiology, food poisonings caused by kudoa septempuctata should be distinguished from those caused by Staphylococcus aureus and bacillus cereus.METHODS: The statistics of food poisoning investigations published by Korea Centers for Disease Control and Prevention in 2013-2015 were reviewed. The characteristics of kudoa septempuctata food poisoning reported by Korea Centers for Disease Control and Prevention were reviewed. Information regarding clinical symptoms or epidemiology was extracted.RESULTS: Total eleven kudoa septempuctata food poisoning cases were analyzed. Food poisonings caused by kudoa septempuctata, Staphylococcus aureus and bacillus cereus have clinical and epidemiological similarities. Forty five percent of food poisoning outbreaks occurred in Korea was concluded as unknown. The food poisoning caused by Staphylococcus aureus and bacillus cereus accounted for 4.5% (50/1,092) of all food poisoning outbreaks in Korea between 2013 and 2015.CONCLUSIONS: This study suggests the possibilities of misdiagnosis in the investigations of food poisoning by Staphylococcus aureus and bacillus cereus with kudoa septempuctata.

Relationship between shipping amounts of olive flounder aquacultured from Jejudo and the reported events of acute food poisoning by Kudoa septempunctata in 2015, South Korea: an ecological study / 한국역학회지

OBJECTIVES: Confirmation of Kudoa septempunctata (K. septempunctata) as the pathogenic agent causing acute food poisoning remains under debate owing to inconsistencies in the reproducibility of experimental evidence. Higher intake of olive flounder infected with K. septempunctata would result in increased diagnosis of food poisoning by K. septempunctata, if the latter was one of the causal agents of acute food poisoning. The aim was to evaluate the relationship between the shipping amount of olive flounder aquacultured from Jejudo and the incidence of K. septempunctata food poisoning in 2015, Korea.METHODS: Data of shipping amounts between March 2014 and February 2016 and of monthly reported events of Kudoa food poisoning were taken from Jejudo Fish-Culture Fisheries Cooperatives and Korea Centers for Disease Control and Prevention, respectively. Non-parametric correlation analyses were conducted.RESULTS: Shipping amounts indicated the seasonal changes according to variation of consumption. Spearman's rho and Kendall's tau-a between the monthly shipping amounts and the reported events in 2015 were 0.39 (p=0.21) and 0.27 (p=0.20), respectively.CONCLUSIONS: An independent relationship was noted between the shipping amount and the reported events, which contrasted with the claim that the virulence of K. septempunctata caused acute food poisoning.

Genomic Analysis of miR-21-3p and Expression Pattern with Target Gene in Olive Flounder

MicroRNAs (miRNAs) act as regulators of gene expression by binding to the 3’ untranslated region (UTR) of target genes. They perform important biological functions in the various species. Among many miRNAs, miR-21-3p is known to serve vital functions in development and apoptosis in olive flounder. Using genomic and bioinformatic tools, evolutionary conservation of miR-21-3p was examined in various species, and expression pattern was analyzed in olive flounder. Conserved sequences (5’-CAGUCG-3’) in numerous species were detected through the stem-loop structure of miR-21-3p. Thus, we analyzed target genes of miR-21-3p. Among them, 3’ UTR region of PPIL2 gene indicated the highest binding affinity with miR-21-3p based on the minimum free energy value. The PPIL2 gene showed high expression levels in testis tissue of the olive flounder, whereas miR-21-3p showed rather ubiquitous expression patterns except in testis tissue, indicating that miR-21-3p seems to control the PPIL2 gene expression in a complementary repression manner in various tissues of olive flounder. Taken together, this current study contributes to infer the target gene candidates for the miR-21-3p using bioinformatics tools. Furthermore, our data offers important information on the relationship between miR-21-3p and target gene for further functional study.

Analysis of Kudoa septempunctata as a cause of foodborne illness and its associated differential diagnosis / 한국역학회지

OBJECTIVES: Recently kudoa septempuctata in olive flounders is suggested as a cause of food poisoning, however whether kudoa septempuctata can affect human gastrointestinal systems is controversial and its pathogenecity remains unclear. In view of the field epidemiology, food poisonings caused by kudoa septempuctata should be distinguished from those caused by Staphylococcus aureus and bacillus cereus. METHODS: The statistics of food poisoning investigations published by Korea Centers for Disease Control and Prevention in 2013-2015 were reviewed. The characteristics of kudoa septempuctata food poisoning reported by Korea Centers for Disease Control and Prevention were reviewed. Information regarding clinical symptoms or epidemiology was extracted. RESULTS: Total eleven kudoa septempuctata food poisoning cases were analyzed. Food poisonings caused by kudoa septempuctata, Staphylococcus aureus and bacillus cereus have clinical and epidemiological similarities. Forty five percent of food poisoning outbreaks occurred in Korea was concluded as unknown. The food poisoning caused by Staphylococcus aureus and bacillus cereus accounted for 4.5% (50/1,092) of all food poisoning outbreaks in Korea between 2013 and 2015. CONCLUSIONS: This study suggests the possibilities of misdiagnosis in the investigations of food poisoning by Staphylococcus aureus and bacillus cereus with kudoa septempuctata.

Relationship between shipping amounts of olive flounder aquacultured from Jejudo and the reported events of acute food poisoning by Kudoa septempunctata in 2015, South Korea: an ecological study / 한국역학회지

OBJECTIVES: Confirmation of Kudoa septempunctata (K. septempunctata) as the pathogenic agent causing acute food poisoning remains under debate owing to inconsistencies in the reproducibility of experimental evidence. Higher intake of olive flounder infected with K. septempunctata would result in increased diagnosis of food poisoning by K. septempunctata, if the latter was one of the causal agents of acute food poisoning. The aim was to evaluate the relationship between the shipping amount of olive flounder aquacultured from Jejudo and the incidence of K. septempunctata food poisoning in 2015, Korea. METHODS: Data of shipping amounts between March 2014 and February 2016 and of monthly reported events of Kudoa food poisoning were taken from Jejudo Fish-Culture Fisheries Cooperatives and Korea Centers for Disease Control and Prevention, respectively. Non-parametric correlation analyses were conducted. RESULTS: Shipping amounts indicated the seasonal changes according to variation of consumption. Spearman's rho and Kendall's tau-a between the monthly shipping amounts and the reported events in 2015 were 0.39 (p=0.21) and 0.27 (p=0.20), respectively. CONCLUSIONS: An independent relationship was noted between the shipping amount and the reported events, which contrasted with the claim that the virulence of K. septempunctata caused acute food poisoning.

IgE Sensitization Patterns to Commonly Consumed Foods Determined by Skin Prick Test in Korean Adults

Offending food allergens can vary with regional preferences in food consumption. In this study, we analysed sensitization rates to commonly consumed foods in Korean adults suspected of having food allergy. One hundred and thirty four subjects underwent a skin prick test (SPT) with 55 food allergens, of which 13 were made by our laboratory and the rest were commercially purchased. Of the 134 patients, 73 (54.5%) were sensitized to one or more food allergens. Sensitization to chrysalis was detected most frequently, at a rate of 25.4%. Sensitization rates to other food allergens were as follows: maize grain (13.4%), shrimp (11.9%), almond (11.1%), wheat flour (8.2%), lobster (8.2%), buckwheat (8.2%), mackerel (5.2%), pollack (5.2%), halibut (4.5%), peanut (4.5%), anchovy (4.4%), squid (3.7%), saury (3.0%), common eel (3.0%), yellow corvina (3.0%), hairtail (2.2%), octopus (2.2%), and others. In addition to well-known food allergens, sensitivity to mackerel, chrysalis, pollack, and halibut, which are popular foods in Korea, was observed at high rates in Korean adults. We suggest that the SPT panel for food allergy in Korea should include these allergens.

Expression of Pleurocidin from winter flounder in Escherichia coli and optimization of culture conditions / 生物工程学报

To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.

Antimicrobial activity of essential oil of Eucalyptus globulus against fish pathogenic bacteria / 한국실험동물학회지

The antibacterial activities of the essential oil of Eucalyptus globulus (EOEG) was determined against 7 fish pathogenic bacteria (Edwardsiella tarda, Streptococcus iniae, S. parauberis, Lactococcus garviae, Vibrio harveyi, V. ichthyoenteri and Photobacterium damselae) obtained from farmed olive flounder. The inhibitory activity was evaluated by three methods: Disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). According to the disc diffusion test, as the concentration of EOEG (5-40 µg) rises, the inhibitory zone increases in size. Compared with amoxicillin, tetracycline and chloramphenicol, EOEG showed similar antibacterial activity. The MIC of EOEG ranged from 7.8 to 125 mg/mL and MBC values ranged from 62 to 250 mg/mL. These results show that EOEG has antimicrobial activity against all seven bacteria, but there was no marked difference between each genus. From these results, it is suggested that EOEG can be used as an antimicrobial agent against fish bacterial diseases in the fish industry.

Genetic and morphological characterisation of a new species of the genus Hysterothylacium (Nematoda) from Paralichthys isosceles Jordan, 1890 (Pisces: Teleostei) of the Neotropical Region, state of Rio de Janeiro, Brazil

Taking into account the difficulties of taxonomic identification of larval anisakid nematodes based on morphological characters, genetic analyses were performed, together with those usually applied, in order to identify anisakid larvae found in the flounder Paralichthys isosceles from the littoral of the state of Rio de Janeiro, Brazil. The analysis of 1,820 larvae revealed a new species, similar to Hysterothylacium MD, Hysterothylacium 2, Hysterothylacium KB and Hysterothylacium sp regarding the absence of the larval tooth, an excretory pore situated below the nerve ring level, and slender lateral alae. Moreover, the new species differs from Hysterothylacium fortalezae and Hysterothylacium reliquens with regard to the number and size of spines present on the tail end and from Hysterothylacium patagonicus by the absence of interlabia. The maximum parsimony and neighbour joining tree topologies based on the 18S ribosomal DNA gene, complete internal transcribed spacer region and cytochrome oxidase 2 (COII) gene demonstrated that the Brazilian larvae belong to Raphidascarididae and represent a unique genetic entity, confirmed as a new Hysterothylacium species. Furthermore, the new species presents COII genetic signatures and shares polymorphisms with Raphidascarididae members. This is the first description of a new anisakid species from Brazil through the integration of morphological and molecular taxonomy data.

Micobacteriose por Mycobacterium marinum em "linguado" Paralichthys orbignyanus e em "barber goby" Elacatinus figaro: diagnóstico histopatológico e imuno-histoquímico / Micobacteriosis by Mycobacterium marinum in Brazilian flounder Paralichthys orbignyanus and barber goby Elacatinus figaro: histopathological and imunohistochemical diagnosis

As micobacterioses afetam várias espécies de peixes, tanto comerciais como ornamentais. É uma enfermidade sistêmica com formação de granulomas, conhecida como micobacteriose de peixes. O desenvolvimento da aquicultura tem aumentado o número de casos com essa enfermidade que muitas vezes é de difícil diagnóstico. Neste trabalho apresentamos dois casos de micobacterioses em Paralichthys orbignyanus e Elacatinus figaro provenientes do Laboratório de Piscicultura Estuarina e Marinha da Universidade Federal do Rio Grande (FURG). As micobactérias de peixes possuem menos ácidos graxos que as de mamíferos, por isso foi utilizado o protocolo de coloração de Fite-Faraco, uma vez que o protocolo de coloração clássico de Ziehl Neelsen pode resultar em falsos negativos. Para determinar o tipo de micobactéria foi utilizada a imuno-histoquímica com a qual se pode diagnosticar a presença de Mycobacterium marinum. A enfermidade pode ser transmitida para o homem e com esse trabalho ressalta-se a necessidade de fazer o diagnóstico correto e implantar medidas de proteção para as pessoas que estão em contato com os peixes infectados.

Mycobacteriosis affects various species of fish, including those cultured with commercial purposes. It is a chronic systemic with granuloma forming disease known as fish mycobacteriosis. In the last few years the interest of fish culture has been increasing due to the reduction of the natural fish stocks. However the intensification of fish culture has collaborated to the development of diseases such as mycobacteriosis, causing losses due to the chronic characteristics and difficult diagnosis. This study is to report two cases of mycobacteriosis observed at "Laboratório de Piscicultura Estuarina e Marinha of the Universidade Federal do Rio Grande (FURG)", one for Brazilian flounder Paralichthys orbignyanus and another for barber goby Elacatinus figaro. The Fite-Faraco staining protocol was used. Membranes of mycobacterial cells contain a waxy substance composed of mycolic acids. These are ß-hydroxy carboxylic acids with chain lengths of up to 90 carbon atoms. The property of acid fastness is related to the carbon chain length of the mycolic acid found in any particular species. The fish mycobacterium is much less acid and alcohol fast than the tubercle bacillus and the classic Ziehl-Neelsen Staining Protocol may give false results. The presence of mycobacterial antigen lesions was studied by immunohistological methods using monoclonal antibodies to Mycobacterium marinum. Humans may be infected by these bacteria and develop cutaneous granuloma. In this paper the necessity is stressed for a correct diagnosis of the disease, contributing to the prevention of its establishment in aquatic facilities, and thus, preserving cultured fish as well as human beings from infection.

External Application of Fermented Olive Flounder (Paralichthys olivaceus) Oil Alleviates Inflammatory Responses in 2,4-Dinitrochlorobenzene-induced Atopic Dermatitis Mouse Model

Allergic skin inflammation such as atopic dermatitis (AD) is characterized by edema and infiltration with various inflammatory cells such as mast cells, basophils, eosinophils and T cells. Thymic stromal lymphopoietin (TSLP) is produced mainly by epidermal keratinocytes, as well as dermal fibroblasts and mast cells in the skin lesions of AD. Omega-3 polyunsaturated fatty acids in fish oil can reduce inflammation in allergic patients. Fermentation has a tremendous capacity to transform chemical structures. The anti-inflammatory effects of fish oil have been described in many diseases, but the beneficial effects by which fermented olive flounder oil (FOF) modulates the allergic response is poorly understood. In this study, we produced FOF and tested its ability to suppress the various allergic inflammatory responses. The ability of FOF to modulate the immune system was investigated using a mouse model of AD. The FOF-treated group showed significantly decreased immunoglobulin E (IgE) and histamine in serum. Also, the increased TSLP expression was significantly inhibited in the FOF group; the FOF-treated group was not appreciably different from the hydrocort cream treatment group. In addition, FOF treatment resulted in a smaller spleen size with reduced the thickness and length compared to the induction group. Splenocytes from mice treated with FOF produced significantly less IFN-gamma, IL-4, T-box transcription factor (T-bet) and GATA binding protein 3 (GATA3) expression compared with the induction group. These results suggest that FOF may be effective in treating the allergic symptoms of AD. 5.

Variações ontogênicas e sazonais na dieta de Etropus crossotus Jordan e Gilbert, 1882 (Paralichthydae) na baía de Ubatuba-Enseada, Estado de Santa Catarina, Brasil / Ongenic and seasonal variations in the diet of Etropus crossotus Jordan and Gilbert, 1882 (Paralichthydae) in the bay of Ubatuba-Enseada, State of Santa Catarina, Brazil

Foram investigadas as variações ontogênicas e sazonais na dieta de Etropus crossotus coletados na baía de Ubatuba-Enseada, Estado de Santa Catarina, Brasil. Redes de arrasto com porta foram utilizadas para as coletas mensais, e 60 exemplares foram coletados entre outubro de 2003 e setembro de 2004. Maior similaridade entre os Índices de Importância Alimentar foi observada entre a primavera e o outono. Crustáceos decápodes representaram 50% dos itens alimentares durante a primavera e outono, e crustáceos anfípodes foram os itens mais representativos no inverno. Maior similaridade entre os IAi foi observada entre as classes 1, 2 e 4, pela alta representatividade de Decapoda, e entre as classes 5, 6 e 7, em função da representatividade de Decapoda e Amphipoda. Os resultados obtidos foram similares a outros estudos realizados na região costeira Sul do Brasil.

Ontogenetic and seasonal changes in the diet of Etropus crossotus sampled at Ubatuba -Enseada Bay in Santa Catarina State, Brazil were investigated. A trawl net was used for the surveys, and 60 fish were collected monthly during October (2003) and September (2004). The highest similarity of the Alimentary Importance Index (AII) was observed between spring and autumn. Crustacea Decapoda represented 50% of the feeding items during spring and fall, and Crustacea Amphipoda were the most representative item in winter. The highest AII similarity was observed among length classes 1, 2 and 4, which was related to Decapoda representation, and among length classes 5, 6 and 7, because of Decapoda and Amphipoda importance. Results were similar to other studies in the southern coast of Brazil.