Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report / Myxobolus spp. (Cnidaria: Myxobolidae) no sangue circulante de peixes dos estados de Goiás e Mato Grosso, Brasil: relato de caso

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

A mixosporidiose é considerada uma doença infecciosa causada por mixozoários pertentences ao Filo Cnidaria, Classe Myxosporea e Ordem Bivalvulida, considerados parasitos comuns de peixes de água doce e salgada, parasitando vários órgãos, principalmente as brânquias. No presente estudo, 49 espécimes de peixes pertencentes a oito gêneros: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus and Serrasalmus foram coletados e extensões sanguíneas foram feitas com a finalidade de encontrar hemoparasitos. Entretanto, mixoesporos foram observados em cinco (10.20%) espécimes de peixes, pertencentes aos gêneros Tetragonopterus, Myleus e Pygocentrus. Dois morfotipos de Myxobolus spp. foram identificados parasitando esses peixes. Normalmente, as investigações sobre mixozoários em peixes é realizada com a procura de plasmódios ou cistos nos órgãos dos peixes e observação da cavidade de órgãos. Porém, esse trabalho ressalta a importância de se examinar também o sangue desses animais, já que esses parasitos podem ocasionar patogenias severas em peixes. Sendo assim, a análise sanguínea pode proporcionar controle sanatório preventivo para peixes comerciais e evitar perdas econômicas.

Las medusas en los balnearios de la provincia de Buenos Aires / The jellyfish species in the coastal waters of Buenos Aires province

Resumen Las medusas son organismos mayoritariamente marinos pertenecientes al grupo de los cnidarios, los cuales se caracterizan por presentar células urticantes especializadas, los cnidocitos. Si bien, todas las medusas son potencialmente venenosas y el grado de toxicidad depende de la especie, sus efectos sobre los humanos varían desde reacciones locales leves hasta reacciones atópicas-anafilácticas graves, llegando incluso a la muerte de la víctima. Mundialmente se conocen numerosas especies de medusas causantes de envenenamiento a humanos, tales como la avispa de mar (Chironex fleckeri), el sifonóforo carabela portuguesa (Physalia physalis) o el hidrocoral de fuego (Millepora spp.). En Argentina, tres especies de medusas revisten de im portancia clínica epidemiológica debido a su poder urticante: las hidromedusas Liriope tetraphylla y Olindias sambaquiensis, así como la escifomedusa Chrysaora lactea. Estas especies presentan sus mayores abundancias en el verano en las costas bonae renses, coincidiendo con la presencia de turistas durante el periodo vacacional. Sus afectaciones varían de leves a moderadas, registrándose desde parestesias y ardor con dermatitis, prurito, edemas y eritemas. Una infinidad de "remedios caseros" se conocen para remediar los efectos de las picaduras de medusas, sin embargo, la mayoría han resultado ineficaces y perjudiciales. Lo más recomendable es evitar frotar y lavar la zona afectada con agua dulce o aplicar hielo para tratar de contrarrestar el ardor y acudir lo antes posible al centro de salud más cercano.

Abstract Medusae are mainly a marine group belonged to cnidarians, which are characterized by specialized stinging cells, cnidocyts. Although all medusae are potentially poisonous, their toxicity depends on the species and the effects on humans vary- ing from mild local reactions to severe atopic-anaphylactic reactions, even the death of the victim. Numerous species of cnidar ians are known worldwide to affect humans, such as the sea wasp {Chironex fleckeri), the siphonophore portuguese man-of-war (Physaliaphysalis) or the fire-coral (Millepora spp.). In Argentina, three species of medusae are known with clinical epidemiological importance due to their stinging power: the hydromedusae Liriopetetraphylla and Olindias sambaquiensis, as well as the scypho- medusae Chrysaora lactea. These species have their highest abundances in the summer on Buenos Aires coasts, coinciding with the presence of tourists during the summer vacations. Its affectations vary from mild to moderate, registering from paresthesias and burning with dermatitis, itching, edemas, and erythema. An infinity of "home remedies" are known formedusae stings, how- ever, most of them have proven ineffective and harmful. It is best to avoid rubbing and washing the affected area with fresh water or applying ice to try to counteract the burning in the region and to go the health center as soon as possible.

Insights into individual variations in nematocyst venoms from the giant jellyfish Nemopilema nomurai in the Yellow Sea.

The giant jellyfish, Nemopilema nomurai, is widely distributed from the Eastern China Sea to the northern part of the Yellow Sea and has resulted in numerous hospitalizations in coastal areas of China, especially in Northern China. Our previous studies have revealed sting-related proteins in the venom of the jellyfish N. nomurai by using experimental and omics-based approaches; however, the variable symptoms of patients who have been stung by N. nomurai are not fully understood. This limited knowledge led to an examination of whether intraspecific variations occur in the venom of different N. nomurai. In the present study, 13 specimens of N. nomurai were collected from the Yellow Sea, and their venom was characterized by profiling differences in biochemical properties and biological activities. SDS-PAGE analysis presented recognizable differences in the number, intensity and presence of some protein bands. Moreover, enzymatic assays revealed considerable quantitative variations in metalloproteinase activity and PLA2-like activity. In particular, zymography assays of proteases demonstrated the general presence of abundant metalloproteinases in jellyfish nematocyst venom; however, the catalytic activities varied greatly among some specific metalloproteinases in the 28-46 kDa or 57-83 kDa range. Hemolytic assays using sheep erythrocytes suggested a predominant variance in the toxicities of different individual jellyfish venoms, with the difference between the most hemolytic and the least hemolytic venom as large as 77-fold. The current data suggested remarkable variations in the nematocyst venoms of individual N. nomurai jellyfish. These observations will provide a new understanding of the clinical manifestations induced by N. nomurai jellyfish stings and will therefore have important implications for preventing and treating jellyfish envenomations.

Unique Diversity of Sting-Related Toxins Based on Transcriptomic and Proteomic Analysis of the Jellyfish Cyanea capillata and Nemopilema nomurai (Cnidaria: Scyphozoa).

The scyphozoan jellyfish Cyanea capillata and Nemopilema nomurai are common blooming species in China. They possess heterogeneous nematocysts and produce various types of venom that can elicit diverse sting symptoms in humans. However, the differences in venom composition between the two species remain unclear. In this study, a combined transcriptomic and proteomic approach was used to identify and compare putative toxins in penetrant nematocysts isolated from C. capillata and N. nomurai. A total of 53 and 69 putative toxins were identified in C. capillata nematocyst venom (CnV) and N. nomurai nematocyst venom (NnV), respectively. These sting-related toxins from both CnV and NnV could be grouped into 10 functional categories, including proteinases, phospholipases, neurotoxins, cysteine-rich secretory proteins (CRISPs), lectins, pore-forming toxins (PFTs), protease inhibitors, ion channel inhibitors, insecticidal components, and other toxins, but the constituent ratio of each toxin category varied between CnV and NnV. Metalloproteinases, proteases, and pore-forming toxins were predominant in NnV, representing 27.5%, 18.8%, and 8.7% of the identified venom proteins, respectively, while phospholipases, neurotoxins, and proteases were the top three identified venom proteins in CnV, accounting for 22.6%, 17.0%, and 11.3%, respectively. Our findings provide comprehensive information on the molecular diversity of toxins from two common blooming and stinging species of jellyfish in China. Furthermore, the results reveal a possible relationship between venom composition and sting consequences, guiding the development of effective treatments for different jellyfish stings.

Organelle survival in a foreign organism: Hydra nematocysts in the flatworm Microstomum lineare.

Nematocysts are characteristic organelles of the phylum cnidaria. They are designated kleptocnidae when sequestered in animals that feed on cnidaria. Kleptocnidae are known for more than a century. Nevertheless it is still enigmatic how selected nematocyst types survive in the predator and how they reach their final destination in the foreign body. In the free-living Platyhelminth Microstomum lineare the fate of nematocysts of the prey Hydra oligactis was analyzed at the ultrastructural level and by fluorescence microscopy using hydra polyps that had been stained in vivo with the fluorescent dyes TROMI and TRITC. M. lineare digested hydra tissue in its intestine within 30 min and all nematocyst types were phagocytosed without adherent cytoplasm by intestinal cnidophagocytes. Desmoneme and isorhiza nematocysts were digested whereas cnidophagocytes containing the venom-loaded stenotele nematocysts started to migrate out of the intestinal epithelia through the parenchyma to the epidermis thereby traversing the subintestinal and subepidermal muscle layer. Within one to two days, M. lineare began to form a muscle layer basolateral around epidermal cnidophagocytes. Epidermal stenoteles survived in M. lineare for at least four weeks. The ability of epidermal stenotele nematocysts to discharge suggest that this hydra organelle preserved its physiological properties in the new host.

Effects of freezing, drying, ultraviolet irradiation, chlorine, and quaternary ammonium treatments on the infectivity of myxospores of Myxobolus cerebralis for Tubifex tubifex.

The effects of freezing, drying, ultraviolet irradiation (UV), chlorine, and a quaternary ammonium compound on the infectivity of the myxospore stage of Myxobolus cerebralis (the causative agent of whirling disease) for Tubifex tubifex were examined in a series of laboratory trials. Freezing at either -20 degrees C or -80 degrees C for a period of 7 d or 2 months eliminated infectivity as assessed by the absence of production of the actinospore stage (triactinomyxons [TAMs]) from T. tubifex cultures inoculated with treated myxospores over a 4-5-month period. Myxospores retained infectivity when held in well water at 5 degrees C or 22 degrees C for 7 d and when held at 4 degrees C or 10 degrees C d for 2 months. In contrast, no TAMs were produced from T. tubifex cultures inoculated with myxospores held at 20 degrees C for 2 months. Drying of myxospores eliminated any evidence of infectivity for T. tubifex. Doses of UV from 40 to 480 mJ/cm2 were all effective for inactivating myxospores of M. cerebralis, although a few TAMs were detected in one replicate T. tubifex culture at 240 mJ/cm2 and in one replicate culture at 480 mJ/cm2. Treatments of myxospores with chlorine bleach at active concentrations of at least 500 mg/L for 15 min largely inactivated myxospore infectivity for T. tubifex. Likewise, there was no evidence of TAMs produced by T. tubifex inoculated with myxospores treated with alkyl dimethyl benzyl ammonium chloride (ADBAC) at 1,500 mg/L for 10 min. Treatments of myxospores with 1,000-mg/L ADBAC for 10 min reduced TAM production in T. tubifex cultures sevenfold relative to that in cultures inoculated with an equal number of untreated myxospores. These results indicate that myxospores of M. cerebralis demonstrate a selective rather than broad resistance to selected physical and chemical treatments, and this selective resistance is consistent with conditions that myxospores are likely to experience in nature.

A real-time polymerase chain reaction assay for the detection of the myxozoan parasite Henneguya ictaluri in channel catfish.

Proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya ictaluri, is the most prevalent parasitic infection affecting commercial channel catfish (Ictalurus punctatus) aquaculture. There are currently no effective chemotherapeutic or biological control measures for PGD, which often peaks during the spring and fall when water temperatures are between 16-25 degrees C. The current diagnostic techniques of gross examination of gill clip wet mounts and histopathology are subject to false-negatives during the early stages of infection, and the quantifiable nature of end-point polymerase chain reaction (PCR) is subjective. Consequently, a rapid and more sensitive quantitative real-time PCR assay was developed for the detection of H. ictaluri during the early stages of infection in channel catfish. A 23 base-pair TaqMan probe was designed based on previously published H. ictaluri PCR protocols. The sensitivity of the assay was the equivalent of a single H. ictaluri actinospore, and in a pond challenge study, quantitative real-time PCR proved to be more sensitive than gross examination, microscopic examination of gill clip wet mounts, and histopathologic examination of gill tissue sections. Future applications of this assay will focus on developing methodologies to be used in conjunction with current pond-monitoring protocols to evaluate potential treatments and better manage this significant seasonal disease.

Fish immune response to Myxozoan parasites.

Myxozoan parasites are responsible for important economic losses among fisheries and aquaculture industries, and hence the high interest in studying the immune response of fish against them. The most important data available concerning the immune response of fish against myxosporeans are reviewed, with emphasis on the different innate and adaptive immune mechanisms, their relationship with natural and acquired resistance and the strategies to control and prevent myxosporoses. Cellular effectors (lymphocytes, granulocytes, phagocytes, non-specific cytotoxic cells, rodlet cells) and humoral factors (lysozyme, peroxidades, antiproteases, complement, specific antibodies) have been examined for several myxosporoses, and some immune relevant genes have been studied. This information will be crucial for the future development of vaccines and other preventive strategies such as immunomodulation and selection of disease-resistant strains

Picadura de medusa / Jellyfish swing

En los últimos años, se ha triplicado la presencia de medusas en nuestros mares, sobre todo en las costas mediterráneas. En el verano de 2005 se ha intensificado el número de picaduras o envenenamientos causados por estos animales marinos. Se piensa, aunque no existe evidencia científica suficiente, que dicho aumento en el número de casos, así como en la gravedad e intensidad de las lesiones, están relacionados con el incremento de la temperatura de las aguas, desde las costas catalanas hasta Cádiz, debido al menor aflujo de agua de los ríos a causa de la importante sequía que sufre nuestro país. En relación con años anteriores, el número de casos de picadura de medusa se ha triplicado. En las costas de Granada se han producido durante el verano de 2005 más de 1.400 asistencias por esta picadura cada fin de semana, lo que ha supuesto cerca del 10% de todas las urgencias atendidas en la comarca. Se presenta, a continuación, el caso de un varón de 16 años que, mientras se bañaba en las playas de Ibiza, sufrió de forma brusca una sensación de quemazón en la región dorsal izquierda, secundaria a picadura de medusa

In recent years, the presence of jellyfish in our seas has tripled, especially in Mediterranean coastal waters. This past summer, the number of stings, or envenomations, caused by these marine animals has grown. Although there is not enough scientific evidence to prove it, it is thought that this increase in the number of cases, as well as the seriousness and severity of the lesions, is related to the rise in the temperature of the coastal waters from Catalonia to Cadiz. This is due to the reduced affluence of water from the rivers owing to the severe drought to which Spain is being subjected. The number of cases has tripled with respect to previous years. During the summer of 2005, in the coastal waters of Granada, more than 1400 jellyfish stings were treated every weekend, an incidence that constitutes nearly 10% of all the requests for emergency care in that region. We report the case of a 16-year-old boy who, while bathing at a beach in Ibiza, experienced a sudden burning sensation in the left dorsal region caused by the sting of a jellyfish

Histopathological methods for the investigation of microbial communities associated with disease lesions in reef corals.

AIMS: To determine the spatial structure of microbial communities associated with disease lesions of reef corals (Scleractinia). METHODS AND RESULTS: Agarose pre-embedding preserved the structure of the disease lesion and surrounding tissues prior to demineralization of the carbonate exoskeleton and embedding in resin. Fluorescence in situ hybridization (FISH) was used to localize bacteria in the lesions of various diseases. CONCLUSIONS: The techniques successfully preserved the in situ spatial structure of degenerated coral tissues. In one case (white plague disease), significant bacterial populations were found only in fragmented remnants of degenerated coral tissues at the lesion boundary that would not have been detected using conventional histopathological techniques. SIGNIFICANCE AND IMPACT OF THE STUDY: Determining the composition, spatial structure and dynamics of microbial communities within the disease lesions is necessary to understand the process of disease progression. The methods described may be applicable to a wide range of diseases involving necrotic lesion formation and requiring extensive tissue processing, such as skeleton demineralization.

Fatty acid composition of bacteria associated with the toxic dinoflagellate Ostreopsis lenticularis and with Caribbean Palythoa species.

The fatty acid composition of a Pseudomonas sp. (Alteromonas) and its host, the dinoflagellate Ostreopsis lenticularis, vectors in ciguatera fish poisoning, has been studied. The major fatty acids in O. lenticularis were 16:0, 20:5n-3, and 22:6n-3, but 18:2n-6, 18:3n-3, and 18:n-3 were also identified. In contrast to other dinoflagellates, 1 8:5n-3 was not detected in O. lenticularis. Even-chain fatty acids such as 9-16:1, 11-18:1, and 13-20:1 predominated in the Pseudomonas sp. from O. lenticularis, but 1 6-20% of (E)-11-methyl-12-octadecenoic acid was also identified. The chirality of the latter was confirmed by total synthesis (28% overall yield) starting from oxacyclotridecan-2-one. The fatty acid compositions of two other Pseudomonas species, from the palytoxin-producing zoanthids Palythoa mamillosa and P. caribdea, were also studied and were similar to that of the Pseudomonas sp. from O. lenticularis. The possibility of using some of these fatty acids as chemotaxonomic lipids in identifying marine animals that consume toxic dinoflagellates or zoanthids is discussed.

HPLC separation of toxic fraction components extracted from planktonic and benthic Cnidaria.

HPLC separation of crude extract components derived from nematocysts and extranematocystic tissues of macroplanktonic jellyfish Aequorea aequorea and Rhizostoma pulmo and benthic sea-anemones Actinia equina and Anemonia sulcata was carried out by different columns. A satisfactory peak separation was obtained analyzing the toxin of Rhizostoma pulmo by cationic and C18 columns. Low molecular weight fragments were separated by C18 column and U.V. monitored varying pH values and obtaining the displacement of significant peaks. Clear differences between chromatographic plots concerning planktonic and benthic species was evidenced by anionic column; this result can point out a clear ecological analogy between species living in the same environment and a similar toxin biosynthesis, due to selective actions related to both the environment and the phylogenetic relationships; these organisms could have developed similar mechanisms useful to tackle the environment.

The neurologic effects of noxious marine creatures.

The concept of the sea as a source of noxious agents is perhaps not a familiar one to clinical neurologists, judging by the lack of reference to these agents in standard textbooks. Chemical, physiologic, and pharmacologic laboratories are increasingly investigating the properties of marine toxins, finding in them compounds with interesting and novel structures or unusual physiologic effects. Such substances are seen as possible agents for biologic and, more particularly, physiologic research, and as possible sources of new pharmaceuticals. These include hormone-like substances and antiviral or antitumor agents. Despite these specialized developments, which are in large measure a consequence of the technological advances of the present century, the clinician is at times directly concerned with the effects of marine toxic substances. For example, in Japan, puffer fish or tetrodotoxic poisoning is one of the major causes of deaths from food poisoning. Another marine toxin that has caused many explosive outbreaks of food poisoning. with many deaths in various parts of the world, comes from clams or mussels. This toxin, saxitoxin, is produced by species of marine protozoa including Gonyaulax, and is concentrated in filter-feeding molluscs. These two examples were of significant interest in medicine long before the technologic developments of the twentieth century. In the last few decades, entirely new problems of marine intoxication have arisen as a result of marine pollution from the disposal of industrial wastes in the sea. The most striking example of a man-made marine intoxication has been the outbreak of Minamata disease. In Minamata, Japan, the disposal of mercury-contaminated industrial wastes from a plastics factory into an enclosed bay, followed by human consumption of the contaminated fishes, crabs, or shellfish, led to many instances of acute cerebral degeneration. With the increasing exploration of the sea for both pleasure and economic exploitation, which is a feature of the second half of the twentieth century, it may be expected that the frequency and variety of human intoxications by marine creatures will be increased. This chapter reviews the neurologic effects of noxious substances of marine biologic origin. The subject is now developing so rapidly that overall surveys, such as this, of the general animal life of theocens will soon be beyond the scope of a single review. Nevertheless, it is hoped that the references given will enable the interested reader to pursue particular aspects further.