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This work investigated the occurrence of Perkinsus sp. in clam Anomalocardia brasiliana, oyster Crassostrea sp. and mussel Mytella falcata from the Jaguaribe River estuary, northeastern Brazil. The collection of clam (N = 300), oysters (N = 300) and mussels (N = 300) were carried out in the estuary of the Jaguaribe River, Ceará, in March and April (rainy season) and October (dry season) in 2017. The mollusks were measured in their major axis, open, and had their tissues submitted to tissue incubation techniques in Ray’s fluid thioglycollate medium (RFTM), histology, real-time polymerase chain reaction (qPCR), PCR and sequencing. The RFTM assays showed Perkinsus sp. infecting the three mollusks investigated. The prevalence of infected clams was 1.33% in both sampling periods, oysters ranged from 2.66 (rainy season) to 8% (dry period), and mussels from 0% (dry period) to 51.33% (rainy season). The intensity of infection was very light to light in clams, very soft to severe in oysters and very soft to moderate in mussels. Histological analyses showed cells of Perkinsus sp. infecting the gills and connective tissue around the digestive gland of some individuals. The qPCR generated amplicons in all positive samples in RFTM, confirming the presence of Perkinsus sp., while the sequencing evidenced high similarity (99%) with the species P. beihaiensis. In conclusion, the results obtained contribute to increasing knowledge about the occurrence of Perkinsus sp. in bivalve mollusks from northeastern Brazil. This work investigated the occurrence of Perkinsus sp. in clam Anomalocardia brasiliana, oyster Crassostrea sp. and mussel Mytella falcata from the Jaguaribe River estuary, northeastern Brazil. The collection of clam (N = 300), oysters (N = 300) and mussels (N = 300) were carried out in the estuary of the Jaguaribe River, Ceará, in March and April (rainy season) and October (dry season) in 2017. The mollusks were measured in their major axis, open, and had their tissues submitted to tissue incubation techniques in Ray’s fluid thioglycollate medium (RFTM), histology, real-time polymerase chain reaction (qPCR), PCR and sequencing. The RFTM assays showed Perkinsus sp. infecting the three mollusks investigated. The prevalence of infected clams was 1.33% in both sampling periods, oysters ranged from 2.66 (rainy season) to 8% (dry period), and mussels from 0% (dry period) to 51.33% (rainy season). The intensity of infection was very light to light in clams, very soft to severe in oysters and very soft to moderate in mussels. Histological analyses showed cells of Perkinsus sp. infecting the gills and connective tissue around the digestive gland of some individuals. The qPCR generated amplicons in all positive samples in RFTM, confirming the presence of Perkinsus sp., while the sequencing evidenced high similarity (99%) with the species P. beihaiensis. In conclusion, the results obtained contribute to increasing knowledge about the occurrence of Perkinsus sp. in bivalve mollusks from northeastern Brazil.

Vibrio parahaemolyticus is a potentially pathogenic bacterium that occurs naturally in estuarine environments worldwide. This research aimed to investigate the occurrence of V. parahaemolyticus in estuarine environments and determine the virulence profile in an aquaculture environment by molecular techniques and conventional microbiological methods. Sampling was conducted in four estuaries in the State of Ceará (Pacoti, Choró, Pirangi and Jaguaribe), Brazil, between January and April 2009. The analysis included 64 samples of water (n=32) and sediment (n=32) collected from the estuaries. The samples yielded 64 isolates suspected to be V. parahaemolyticus. The isolates were submitted to biochemical identification using a dichotomous key and PCR for the detection of the species-specific tlh gene. Virulence was assessed by testing for urea hydrolysis and β-hemolysis in erythrocytes (Kanagawa phenomenon) and simultaneous detection of the tdh and trh genes. All but one of the isolates (63/64) were confirmed to be V. parahaemolyticus by genotypic detection of tlh gene. The tdhand trh genes were detected in 57 and 19 isolates, respectively. The Kanagawa test was positive for 51 isolates. Only one isolate was positive for urease. The incidence of tdh/trh-positivity was very high in isolates recovered from the environment. The present study demonstrates the need to increase knowledge of the ecology and pathogeny of V. parahaemolyticus

This study investigatedPerkinsusspp. infectingCrassostrea rhizophoraefrom the Jaguaribe River estuary,Ceará, Brazil. Fragments of gills and rectum of the oysters (n = 150) were incubated in Ray’s fluid thiogly-collate medium (RFTM). GenusPerkinsus-specific PerkITS85/750 PCR assays were performed and theiramplicons were sequenced by the Sanger method. The RFTM assays confirmedPerkinsusspp. Thesequencing of the amplified fragments from the rDNA internal transcribed spacers (ITS) ofPerkinsusspp. confirmedPerkinsus chesapeaki. Neighbor-Joining analyzes placeP. chesapeakiidentified in this studyin a well-supported clade with other isolates of the same species. This is the first record ofP. chesapeakiinfectingC. rhizophoraein South America.Ó2016 Elsevier Inc. All rights reserved.1. IntroductionThe estuary of the Jaguaribe River (Ceará State, Northeast Brazil)has great importance both from an ecological point of view, as wellas for supporting fisherman communities, as a mollusk collectingsite for consumption and marketing. Mollusks from natural envi-ronments and also from cultivation can be affected by differentgroups of pathogens, including protozoa belonging to the genusPerkinsus. This genus currently has seven accepted species. Amongthem,P. marinusandP. olsenirequire mandatory reporting to theWorld Organization for Animal Health, due to the risk that theyrepresent for mollusk populations (Villalba et al., 2004; Choi andPark, 2010). In Brazil, studies on perkinsosis have attracted atten-tion since the first record ofPerkinsussp. was made inCrassostrearhizophoraefrom the Pacoti River estuary, Ceará (Sabry et al.,2009).Sabry et al. (2013)confirmed that the species ofPerkinsusaffecting oysters in that estuary wasP. beihaiensis. In this sameyear, the first South American records were published ofP. marinusinfectingC. rhizophoraefrom the estuary of Paraíba River (da Silvaet al., 2013) andPerkinsussp. infecting the same species of oysterfrom the coast of Bahia State (Brandão et al., 2013). Recently,Perkinsussp. andP. marinuswere reported infectingCrassostreagasarfrom Paraíba State (Queiroga et al., 2013, 2015.),P. marinusandP. olseniwere identified inC. gasarfrom the estuary of theSão Francisco River, Sergipe State (da Silva et al., 2014) andP. beihaiensiswas recorded inAnomalocardia brasilianafrom theTimonha River estuary, Ceará State (Ferreira et al., 2015). Thisstudy investigated the presence and prevalence ofPerkinsusspp.inC. rhizophoraefrom the estuary of the Jaguaribe River, Ceará,Brazil.2. Materials and methodsThe oysters were manually collected from the roots ofRhizophorae mangleat low tide in the estuary of the Jaguaribe River(04°27039,2100S, 37°47031,6800W), on the east coast of Ceará State.Collections were made in March 2012 (N = 150) during the rainyseason (January-April) and October (N = 150) of the same year dur-ing the dry season (May-December). The temperature and salinitydata were measured during the sampling. The oysters were trans-ported to the laboratory alive in isothermal boxes. Prior to process-ing, the animals were measured for shell height, according toGaltsoff (1964). Two gill lamellae and the rectum of each oysterwere incubated in Ray’s fluid thioglycollate medium in thepresence of antibiotics and antifungals (penicillin/streptomycinand nystatin) for 7 days in the dark at room temperature.

Endangered angelsharks and a protected dolphin species are illegally traded in Brazil. In this study, we determined prospective molecular markers for detecting these species in the trade of angelshark carcasses and ‘dolphin’ eyeball amulets. We compiled publicly available as well as new and unpublished cytochrome b (cyt b) DNA sequences for species involved in these trades. These sequences were digested in silico using restriction enzymes. We then described prospective polymerase chain reaction (PCR)-restriction fragment length polymorphism markers for distinguishing between protected species and the species whose trader was legally allowed in these two trade groups. The prospective marker for identifying angelshark carcasses consists of cyt b PCR and digestion by BstXI, BsgI, BspMI, BsrDI, and HaeII restriction enzymes. The prospective marker for identifying eyeball amulets consists of cyt b PCR and digestion by ApoI, BtsI, HindII, BsaAI, BplI, and SspI restriction enzymes. This is the first study to deposit in GenBank cyt b sequences for the angelshark species Squatina argentina, Squatina guggenheim, and Squatina occulta. Moreover, the S. argentina haplotype is the first DNA sequence for this species deposited in GenBank

The diatomAsterionellopsis glacialissensu latoforms high-density patches in the surf zone of somesandy beaches worldwide and was until recentlyconsidered a cosmopolitan species. With the recentdescription of four cryptic species, the identity ofspecimens found in these accumulations remainsuncertain. In this study, diatom patches weresampled from two sandy beaches of the Braziliancoast: one tropical (Futuro Beach, 3°S; 38°W) andone subtropical (Cassino Beach, 32°S; 52°W). Finestructure of frustules and the sequencing of threephylogenetic markers revealed the subtropicalstrains to beA. guyunusaeand the tropical strains tobe a new species, here described asAsterionellopsistropicalissp. nov.A. tropicaliswas differentiatedmorphologically by the number of striae in 10lmatthe foot pole and head (39–44; 38–45, respectively),fromA. lenisilicea(46–55; 46–64),A.maritima (46–51; 46–60), andA. thurstonii(42–58; 55–70). Thenumber of striae at the head region of thevalvocopula (10lm) helped to distinguishA. tropicalis(56–62) fromA. guyunusae(61–64), butA. tropicaliswas morphologically undistinguishablefromA. glacialis. The sequence divergence fromother identifiedAsterionellopsisspecies was 13%–16%(Cox1), 11%–12% (5.8S+ITS2) and 2%–6% (RbcL),andA. tropicalisformed a distinct monophyleticclade with high support in all analyzed phylogenetictrees (single or multi-locus). This work will aid inthe understanding of the ecological andphysiological diversity of diatom patches that arekey to the trophic webs of sandy beaches

Um total de 119 indivíduos foi criado em um tanque rede (3,0 m x 3,0 m x 2,7 m) no açude castanhão e realizadas medidas no habitat às dez horas da manhã que se apresentaram conforme consideradas a seguir: temperatura da água variou entre 27,3°C e 28,0°C; oxigênio de 7,9 a 3,0; condutividade de 350 a 342; ORP (potencial de oxidação e redução do ambiente) variou de 100 a 127. Observou-se um bom estado de saúde e de desenvolvimento dos indivíduos no tanque rede ao longo do trabalho, com pesos médios (g) aos 7, 9, 10 e 12 meses de idade de 65,0, 94,1, 125,7 e 111,1, respectivamente, demonstrando um crescimento gradual até os 10 meses, com um declínio subsequente aos 12 meses. O dimorfismo sexual ficou caracterizado aos 10 meses de idade quando os indivíduos, reprodutivamente preparados, foram separados por sexo nas pesagens. Aos 10 e 12 meses as fêmeas apresentaram médias (g) e desvios padrões de 150,4 ± 43,0 e 141,9 ± 31,4 e os machos de 93,6 ± 16,7 e 95,9 ± 31,4, respectivamente. Não foi percebida diferença significativa na proporção 1:1, entre os sexos, mas sim nos pesos médios aos 10 (X2 = 15,2) e aos 12 meses (X2 = 9,0), p < 0,01, com as fêmeas alcançando o desenvolvimento máximo aos dez meses. Os resultados proporcionados por este trabalho mostraram ser possível a produção da curimatã comum em tanques rede, com crescimento e desenvolvimento gonadal pleno, tornando possível a obtenção do “caviar do sertão” e a utilização dos animais na reprodução.

O objetivo deste estudo foi detectar a presença potencial virulência de Vibrio cholerae isolado de estuários do Nordeste do Brasil. Amostras de água e sedimento foram coletadas e inoculadas sobre meio seletivo para víbrios (TCBS) e colônias com características morfológicas de V. cholerae foram isoladas. A identificação fenotípica seguiu chave dicotômica baseada em caraterísticas bioquímicas. Foram empregadas as técnicas de amplificação da polimerase em cadeia (PCR) utilizando o gene ompW e a de multiplex PCR para detecção de genes de virulência (ctx, tcp, zot e rfbO1). Os resultados da identificação das diferentes abordagens foram comparados. Nove cepas de V. cholerae foram identificadas fenotipicamente e cinco confirmadas através da detecção do gene ompW. A chave dicotômica utilizada foi eficiente para a confirmação da espécie. Os quatro estuários analisados apresentaram estirpes de V. cholerae, e nenhuma das cepas isoladas apresentaram genes de virulência. The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

ROMÃO, L DE S. et al. Patógenos em duas espécies de bivalves comercialmente importantes do estuário do Rio Pacoti, Estado do Ceará, Brasil. Arquivos de Ciências do Mar, Fortaleza, v. 47, n. 2, p. 57-63, dez. 2014. In Ceará State’s estuaries, the mangrove oyster, Crassostrea rhizophorae and the cockle Anomalocardia brasiliana are harvested by local communities, representing a very important socioeconomic activity. Whereas the pathogens can cause mortality in mollusk populations, this study aimed to investigate the occurrence of parasites in C. rhizophorae and in A. brasiliana from Pacoti River estuary, Ceará State. During the period from February, 2008 to January, 2009, samples of cockles and oysters (n = 360 per species) were collected monthly for histopathological analyses. The results in cockles showed bacterial colonies rickettsia-type (7.2%), metazoan possibly bucephalids (35.8%), Tylocephalum (31.7%) and metacercariae of trematodes (0.5%). In oysters, it was observed protozoa of the genera Nematopsis (83%), unidentified protozoans (3.9%), Ancistrocoma (7.2%) and the metazoa: Urastoma (0.8%), bucephalids (0.2%) and Tylocephalum (3%). Despite the high prevalence of Nematopsis in oysters, damage was not observed in the tissues of infected animals. In contrast, the infestation by bucephalids in the gonads caused the castration of the host. The results indicate that those mollusk species must be monitored in their habitat so as to keep pace with the evolution of the prevalence and the appearance of new pathogens. Nos estuários do Estado do Ceará, a ostra-do-mangue Crassostrea rhizophorae e o búzio Anomalocardia brasiliana são coletados pela comunidade local, representando uma atividade socioeconômica muito importante. Considerando que diversos patógenos podem causar mortlidade nas populações de moluscos o objetivo do presente trabalho foi investigar a ocorrência de parasitas em C. rhizophorae e A. brasiliana do estuário do Rio Pacoti, Ceará. No período de fevereiro/2008 a janeiro/2009, amostras de búzios e ostras (n = 360/espécie) foram coletadas mensalmente para análises histopatológicas. Os resultados nos búzios evidenciaram colônias bacterianas do tipo rickettsia (7,2%), metazoários possivelmente bucefalídeos (35,8%), Tylocephalum (31,7%) e metacercárias de trematódeos (0,5%). Nas ostras foram observados protozoários do gênero Nematopsis (83%), protozoários não-identificados (3,9%), Ancistrocoma (7,2%) e os metazoários: Urastoma (0,8%), bucefalídeos (0,2%) e Tylocephalum (3%). Apesar da elevada prevalência de Nematopsis nas ostras, não foram observados danos aos tecidos dos animais infectados. Ao contrário, à infestação por bucefalídeos causou a castração da gônada dos hospedeiros. Os resultados obtidos indicam que os moluscos devem ser monitorados em seu habitat, visando acompanhar a evolução das prevalências e o surgimento de novos patógenos.

The majority of fish species reared in captivity presents some form of reproductive dysfunction. In females may appear failures of final maturation of the ova, ovulation and the spawning; while in males occurs a decrease in the production of milt and in its quality. These dysfunctions appear due to the modification of the natural conditions of reproduction of the fish in captivity, originated from the lack of external stimuli needed for induction the pituitary to release of gonadotropins. In overcoming these disturbances have been used the hormones of reproduction for over fifty years, initially as pituitary extracts to stimulate the reproductive processes of spermiation, ovulation induction and spawning. More recently the hormones derived from fish and mammals, such as the carp, salmon and chorionic gonadotropin, and employment of new methods, with the factors releasers of gonadotropins (GnRH), which induce the release of gonadotropin own pituitary decreasing the markest failures in creations in captivity. The development of synthetic agonists of GnRH (GnRHa), highly powerful, consist of the new generations of therapies to hormonal manipulation, creating new possibilities with the use of hormones for the control of reproductive processes in aquaculture. The products developed more recently incorporate the GnRHa in polymeric system release long, with the effects of the hormone extending by extensive periods that last for days and even weeks. These systems eliminate the need for multiple treatments, in addition to induce the development of germ cells and the synchronization of multiple spawns. These different forms of GnRH recently found and their better knowledge certainly will be widely used in therapy for spawning. New strategies must be developed to enable a improve achievement of spawning, possibly with research to correct levels of GnRH rolling stock. A maioria das espécies de peixes criadaem cativeiro apresenta alguma forma de disfunção reprodutiva. Nas fêmeas podem aparecer falhas de maturação final dos óvulos, da ovulação e da desova; enquanto nos machos ocorre uma diminuição na produção do sêmen e na sua qualidade. Essas disfunções surgem em função da modificação das condições naturais de reprodução dos peixes em cativeiro, oriundas da falta dos estímulos externos necessários à indução da pituitária para liberação de gonadotrofinas. Na superação desses distúrbios são utilizados os hormônios da reprodução há mais de cinquenta anos, inicialmente como extratos hipofisários, para estimular os processos reprodutivos da espermiogênese, indução da ovulação e desova. Mais recentemente os hormônios derivados de peixes e de mamíferos, como as da carpa, salmão e coriônica humana, e o emprego de novos métodos, com os fatores liberadores de gonadotrofinas (GnRH), que induzem a liberação da própria gonadotrofina da pituitária diminuindo as falhas observas nas criações em cativeiro. O desenvolvimento de agonistas sintéticos de GnRH (GnRHa), altamente potentes, constituem as novas gerações das terapias de manipulação hormonal, criando novas possibilidades com o uso de hormônios para o controle dos processos reprodutivos na aquicultura. Os produtos desenvolvidos mais recentemente incorporam o GnRHa em sistema polimérico de liberação longa, com os efeitos do hormônio se estendendo por períodos amplos que duram dias e até semanas. Esses sistemas eliminam a necessidade de múltiplos tratamentos, além de induzirem o desenvolvimento das células germinativas e a sincronização de desovas múltiplas. As diferentes formas de GnRH encontradas recentemente e seu melhor conhecimento certamente serão amplamente utilizados na terapia da desova. Novas estratégias devem ser desenvolvidas para possibilitar uma melhorar obtenção da desova, possivelmente com pesquisas que venham corrigir os níveis de GnRH circulante.

The species of migratory fish are characterized by displacement along the rivers in rainy periods to perform their reproduction, seasonally allocated. The females of curimatã common (Prochiloduscearaensis), one of these migratory species, present a perfect synchronism between their physiological conditions and the environment, which enables the ovarian development and the final maturation of the ova. In this moment, they are prepared to carry out the ovulation and spawning. However, while in backwater, the mechanism of reproduction is only triggered by anartificial induction, with the employment of pituitary extracts and specific pituitary hormones. Ovulation was inducted in five females with pituitary extract, using a pituitary per kilogram of body weight in two successive doses: the first having 10% of total concentration and the second 90% of the remaining twelve hours after, it was observed two females spawning. After the ovultation induction, three females did not spawn, and, after being sacrificed, they had ovaries in regression phase, which is a reproductive dysfunction observed during the holding in captivity. The knowledge and understanding of the failures causes observed in reproduction are essential for a profitable intensive exploitation of curimatã common, that presents a great economic regional potential and, if correctly grown, can offer protein of high biological value and nutrition to the consumer. As espécies de peixes migradores caracterizam-se por deslocamento ao longo dos rios nos períodos chuvosos para realizar sua reprodução, de forma sazonalizada. As fêmeas de curimatã comum (Prochiloduscearaensis), uma dessas espécies reofílicas, apresentam um sincronismo perfeito entre suas condições fisiológicas e as do ambiente, o que possibilita o desenvolvimento ovariano e a maturação final dos óvulos. Nesse momento, elas se tornam preparadas para realizar a ovulação e a desova. Contudo, em água parada, o mecanismo de reprodução só é desencadeado através da indução artificial com o emprego de extratos hipofisários e hormônios específicos. Induziu-se a ovulação em cinco fêmeas com extrato hipofisário, utilizando uma hipófise por quilo de peso em duas doses sucessivas: a primeira tendo 10% da concentração total e a segunda os 90% restantes doze horas após, observou-se a desova em duas fêmeas. Após a indução da ovulação, três fêmeas não desovaram e, ao serem sacrificadas, apresentavam ovários em fase de regressão, uma disfunção reprodutiva observada durante a exploração em cativeiro. O conhecimento e o entendimento das causas responsáveis dessas falhas observadas na reprodução são essenciais para uma exploração intensiva rentável da curimatã comum, cuja espécie apresenta um grande potencial econômico regional e, ao ser cultivada adequadamente, pode proporcionar proteína de alto valor biológico e nutricional ao consumidor.

This article documents the addition of 153 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Brassica oleracea, Brycon amazonicus, Dimorphandra wilsonii, Eupallasella percnurus, Helleborus foetidus, Ipomoea purpurea, Phrynops geoffroanus, Prochilodus argenteus, Pyura sp., Sylvia atricapilla, Teratosphaeria suttonii, Trialeurodes vaporariorum and Trypanosoma brucei. These loci were cross-tested on the following species: Dimorphandra coccicinea, Dimorphandra cuprea, Dimorphandra gardneriana, Dimorphandra jorgei, Dimorphandra macrostachya, Dimorphandra mollis, Dimorphandra parviflora and Dimorphandra pennigera. © 2013 Blackwell Publishing Ltd.