Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.

Fauna parasitária de peixes oriundos de pesque-pague do município de Franca, São Paulo, Brasil. I. Protozoários

The present work studied protozoan parasites of cultivated fishes (N = 433) from two feefishing farm situated in Franca, São Paulo, Brazil, during a period of April 1997 through March 1999. Specimens of piauçú Leporinus macrocephalus Garavello & Britski, 1988 (Anostomidae), pacu Piaractus mesopotamicus Holmberg, 1887 (Characidae) carp Cyprinus carpio Linnaeus, 1758 (Cyprinidae), Tillapia rendalli Boulenger, 1896 (Cichlidae), nile-tilapia Oreochromis niloticus Linnaeus, 1758 (Cichlidae), matrinxã Brycon cephalus Günther, 1869 (Characidae) and tambacu hybrid (male of P. mesopotamicus x female of Colossoma macropomum Cuvier, 1818) were collected. The fishes were parasitized with protozoans Ichthyophthirius multifiliis Fouquet, 1876 (Protozoa), Trichodina sp. and Piscinoodinium pillulare (Schaperclaus, 1954), Lom, 1981 (Protozoa). In the cold season (autumn and/or winter) all species of fish were infected with I. multifiliis. Higher susceptibility to Trichodina sp. was observed in L. macrocephalus, C. carpio and P. mesopotamicus compared to tambacu, B. cephalus, T. rendalli and 0. niloticus. It was not observed significant difference (P > 0.05) in the seasonal variation of Trichodina sp. and P. pillulare infection of all species. A great number of P. pillulare without significant difference (P > 0.05) was reported to L. macrocephalus, P. mesopotamicus and tambacu.

Comparative evaluation of the susceptibility of cultivated fishes to the natural infection with myxosporean parasites and tissue changes in the host

O objetivo deste estudo foi avaliar a susceptibilidade de 4 importantes peixes cultivados a parasitos esporozoários. Os peixes foram coletados bimestralmente de um tanque de cultivo, durante 1 ano. Myxobolus colossomatis e Henneguya piaractus foram encontrados nos órgãos internos e brânquias, respectivamente. A incidência de ambos os parasitos foi de 97,3% em pacu (Piaractus mesopotamicus), 33,3% no híbrido tambacu (Piaractus mesopotamicus x Colossoma macropomum), 5,6% em tambaqui (Colossoma macropomum) e 0% em carpa (Cyprinus carpio). Pacu foi o peixe mais susceptível, encontrando-se parasitado 79,2% nas brânquias, 66,7% nos rins e 50% no baço. A análise histopatológica das brânquias mostrou hemorragias, reação inflamatória com células mononucleares, fibroblastos e hiperplasia das células basais e mucosas.

Piscinoodinium pillulare (Schäperclaus, 1954) Lom, 1981 (Dinoflagellida) infection in cultivated freshwater fish from the Northeast region of São Paulo State, Brazil: parasitological and pathological aspects

O Centro de Aqüicultura, Unesp, Jaboticabal, SP, Brasil, recebeu peixes para diagnose, os quais apresentavam aglomeração nas bordas dos viveiros e na entrada da água. Dos 194 casos diagnosticados, 53 apresentavam estruturas brancas circulares ou ovais, imóveis, medindo 162 mm de diâmetro, identificadas como o dinoflagelado Piscinoodinium pillulare. em 34 casos, os parasitos estavam presentes nas brânquias, em 2 casos, no corpo e em 9 casos, em ambos. Dos 53 casos observados, 31 eram o híbrido tambacu; 7, o Piaractus mesopotamicus; 6, o Colossoma macropomum; 5, o Leporinus macrocephalus; 3, o Oreochromis niloticus; e 1, o Prochilodus lineatus. Os peixes apresentaram aumento da produção de muco no corpo e nas brânquias e equimoses no pedúnculo caudal e nos opérculos. As brânquias também apresentaram palidez, congestão e petéquias. A histopatologia revelou a presença de grande número de trofontes situados entre as lamelas secundárias, fixados ou não ao epitélio. As lamelas primárias mostraram hemorragias intersticiais, severa hiperplasia do epitélio e das células caliciformes e infiltrado inflamatório. O presente trabalho é o primeiro relato de P. pillulare no Brasil e enfatiza a importância dos dinoflagelados, que causaram significativas perdas econômicas entre 1995 e 1997.

Relação hepatosomática e esplenosomática em peixes teleósteos de cultivo intensivo

The mean values of hepatosomatic relation (HSR), splenosomatic relation (SSR) and relative condition factor (Kn) in Nile tilapia (Oreochromis nilolicus Trewavas, 1983); pacu (Piaractus mesopolamicus Holmberg, 1887); piauçu (Lepori-nus macrocephalus Garavello & Britski, 1988) and tambacu hybrid (P. mesopotamicus male x Colossoma macropomum Cuvier, 1818 female) are described. The experiment was carried out from feefishing farm situated in Franca, São Paulo State, Brazil. Nile tilapia showed the greatest HSR compared to pacu, piauçu and tambacu. Although, in the last three fishes the relation was similar. The SSR between tilapia and piausu was similar, but smaller than observed in pacu and tambacu. Nevertheless, SSR in pacu and tambacu was different. The hepatic weight/body weigth relation and hepatic weight/body length relation presented positive correlation in ali studied fishes. The splenic weight/body weigth relation and splenic weight/body length relation were not significam (P>0.05) in Nile tilapia, pacu and tambacu, but highly significant in piauçu (PO.01).

Fauna parasitária de peixes oriundos de pesque-pagues do município de Franca, São Paulo, Brasil. II. Metazoários

Metazoan fauna from cultivated fishes (N = 433) in two feefishing farm of Franca, State of São Paulo, Brazil, was studied during a period of April 1997 through March 1999. Specimens of piauçú Leporinus macrocephalus Garavello & Britski, 1988 (Anostomidae), pacu Piaractus mesopotamicus Holmberg, 1887 (Characidae), carp Cyprinus carpio Linnaeus, 1758 (Cyprinidae), Tillapia rendalli Boulenger, 1896 (Cichlidae), nile-tilapia Oreochromis niloticus Linnaeus, 1758 (Cichlidae), matrinxã Brycon cephalus Günther, 1869 (Characidae) and tambacu hybrid (male of P. mesopotamicus x female of Colossoma macropomum Cuvier, 1818) were collected. A total of 29.3% was parasitized by monogenean, 9.7% with crustacean (copepodits and adults of Lernaea cyprinacea Linnaeus, 1758 and Argulus sp.) and 1.6% with myxosporean (Henneguya piaractus Martins & Souza, 1997 and Myxobolus colossomatis Moinar & Békési, 1993).The most susceptible species to myxosporean infestation was P. mesopotamicus. L. macrocephalus and P. mesopotamicus that showed a higher susceptibility to Argulus sp infestation. It was not observed significant difference (P > 0.05) in the seasonal variation of L. cyprinacea infection of all species. L. macrocephalus, P. mesopotamicus and tambacu showed a great number of P. pillulare with no significant difference (P > 0.05) between them. Monogenean infections were observed in all examined species, without significant difference (P > 0.05) in the seasonal variation. Nevertheless, occurrence of monogenean was higher in P. mesopotamicus and lower in Nile-tilapia. By the other hand, parasites number collected from P. mesopotamicus was lower in the winter. Copepodits of L. cyprinacea were found in the gills of P. mesopotamicus, tambacu and L. macrocephalus. However, adults of L. cyprinacea were observed only in the P. mesopotamicus and tambacu.

Myoarchitecture and vasculature of the heart ventricle in some freshwater teleosts

The morphological characteristics of the ventricular myocardium and of coronary vascularization were studied in three freshwater teleost species, Piaractus mesopotamicus, Colossoma macropomum and Clarias gariepinus (African catfish), by correlating their ventricular shapes and swimming habits. In Piaractus mesopotamicus and Colossoma macropomum, species with highly active swimming habits, the cardiac ventricle showed a pyramidal shape and a richly vascularized myocardium consisting of an outer compact layer and inner spongy layer. In Clarias gariepinus, a less active species, we observed a saccular ventricle with a mixed myocardium and coronary arteries, in contrast to the ventricular structure of other species described in the literature.

Interspecific fish hybrids in Brazil: management of genetic resources for sustainable use

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular diagnostic methods for identifying Serrasalmid fish (Pacu, Pirapitinga, and Tambaqui) and their hybrids in the Brazilian aquaculture industry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efectos del parasitismo sobre el tejido branquial en peces cultivados: estudio parasitologico e histopatologico

One hundred four out of 225 diagnosticated cases were myxosporidian, monogenean, Ichthyophthirius multifiliis Fouquet, 1876 and bacterial diseases in Piaractus mesopotamicus Holmberg, 1887 (pacu), Colossoma macropomum Cuvier, 1818 (tambaqui) and tambacu, at Aquaculture Center, Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brazil, between 1992 and 1995. The gills were fixed in 10% buffered formalin solution to posterior histologic routine. It was observed Henneguya sp. cysts into the capillaries of the secondary lamellae, encapsulated by the respiratory epithelium cells. It provoked adherence of the adjacent lamellae, hyperplasia, congestion, oedema and epithelium displacement. Monogeneans infestations by Anacanthorus penilabiatus Boeger, Husak & Martins, 1995 and Ancyrocephalinae sub-family caused light inflammatory reaction and hyperplasia. In severe infestations was observed hyperplasia of primary lamellae, necrosis, oedema, respiratory epithelium displacement, ruptured pillar cells and telangiectasis. Response to I. multifiliis was limited to surrounding epithelial cells in young fishes and hyperplasia, necrosis, inflammatory infiltrate and oedema in old fishes. Such lesions iniciate hyperplasic and oedematous process that with inflammation of the parasitic sites, provoked alterations over gases and ions interchange surface and consequently fish metabolism.

Metodologia de infecção experimental e grau de susceptibilidade do híbrido tambacu e Leporinus macrocephalus Garavello & Britski (Osteichthyes, Anostomidae) a quatro inóculos de trofozoítos de Icthyophthirius multifiliis Fouquet (Protozoa, Ciliophora)

Tropical fishes like Leporinus macrocephalus Garavello & Britski, 1988 (piauçu) and the hybrid tambacu (Piaractus mesopotamicus Holmberg, 1887 pacu male x Colossoma macropomum Cuvier, 1818 tambaqui female) were experimentally infected with four inocula of trophonts of Ichthyophthirius multifiliis Fouquet, 1876 (Protozoa, Ciliophora) and the infection level was evaluated. Each one of twelve experimental units consisted of a tank with 200 liters of water and two specimens of each kind of fish. The following treatments were designed to each tank, in triplicate: 8, 16,32 and 64 trophonts. After 8 days of exposure the fishes were killed and the number of trophonts was counted at a stereoscopic microscope. This assay showed to be feasible at tropical conditions and was able to show that tambacu was more susceptiblc than piauçu.

Gill chemoreceptors and cardio-respiratory reflexes in the neotropical teleost pacu, Piaractus mesopotamicus

This study examined the location and distribution of O-2 chemoreceptors involved in cardio-respiratory responses to hypoxia in the neotropical teleost, the pacu (Piaractus mesopotamicus). Intact fish and fish experiencing progressive gill denervation by selective transection of cranial nerves IX and X were exposed to gradual hypoxia and submitted to intrabuccal and intravenous injections of NaCN while their heart rate, ventilation rate and ventilation amplitude were measured. The chemoreceptors producing reflex bradycardia were confined to, but distributed along all gill arches, and were sensitive to O-2 levels in the water and the blood. Ventilatory responses to all stimuli, though modified, continued following gill denervation, however, indicating the presence of internally and externally oriented receptors along all gill arches and either in the pseudobranch or at extra-branchial sites. Chemoreceptors located on the first pair of gill arches and innervated by the glossopharyngeal nerve appeared to attenuate the cardiac and respiratory responses to hypoxia. The data indicate that the location and distribution of cardio-respiratory O-2 receptors are not identical to those in tambaqui (Colossoma macropomum) despite their similar habitats and close phylogenetic lineage, although the differences between the two species could reduce to nothing more than the presence or absence of the pseudobranch.

The effect of ions and adenosine nucleotides on the activity of lactate dehydrogenase from the epaxial muscle of fish

Lactate dehydrogenase was partially purified from the epaxial muscle of Piaractus mesopotamicus (pacu) and its hybrid Piaractus mesopotamicus x Colossoma macropomus (tambacu). This preparation was used for kinetic studies carried out at pH 6.0 and 7.5. It was also used for the study of the inhibition properties of adenosine nucleotides = ATP, ADP, AMP =, divalent ions Ni2+, Cu2+, Co2+ and the anions oxamate and oxalate.

Effects of food quality on growth and biochemical composition of a calanoid copepod, Argyrodiaptomus furcatus, and its importance as a natural food source for larvae of two tropical fishes

Growth, reproduction and biochemical composition were analyzed for the copepod Argyrodiaptomus furcatus fed on the alga Ankistrodesmus gracilis grown in different media. The ingestion of this copepod by larvae of two species of tropical fishes was also evaluated. The mean peak density of the copepod population was 1369 individuals 1-1 for all four diets used, and the highest was 1387 individuals 1-1 on diet ARV (algae + ration + vitamins). A small copepod, A. furcatus tends to have a short life span. The smallest females did not attain maturity in the shortest time on all diets used. Food quality may play a major role in the dynamics of the biochemical composition of this copepod. Argyrodiaptomus furcatus was a more important food item for larvae of tambaqui (Colossoma macropomum) than of pacu (Piaractus mesopotamicus). However, it made up a large part of the gut contents of larvae of both species.

Distribuição sazonal de parasitos branquiasis em diferentes especies de piexes em pesque-pague do municipio de Guariba-SP, Brasil.

This study was accomplished with the objective of knowing the seasonal distribution of the main species of parasites causes of branchial damages to the fish from fee-fishing at Guariba, State of São Paulo. This city is situated at 21 degrees 15'22'' S, 48 degrees 18'58'' W, in altitude 595 meters. This research was conducted between April, 1997 to March, 1999. From 408 fishes examined, 29.1% of them were infested by the protozoans e metazoans. Leporinus macrocephalus showed the highest parasitism by Trichodina sp., although Piaractus mesopotamicus and Cyprinus carpio were also parasited. Piscinoodinium pillulare infestation were found in L. macrocephalus, P. mesopotamicus, C. carpio e Colossoma macropomum. Monogenetics metazoan have largely been observed in L. macrocephalus, P. mesopotamicus, C. carpio, C. macropomum e cross-bred tambacu. The incidence of this parasite was elevated in the summer for L. macrocephalus, spring for P. mesopotamicus and C. macropomum, and both seasons for cross-bred tambacu. Lernaea cyprinacea copepodes were observed in L. macrocephalus, P. mesopotamicus, C. carpio and cross- breeding tambacu, in spite of the fact that C. carpio has demonstrated high parasitism of adult form. The fish L. macrocephalus, P. mesopotamicus and C. carpio were sponged intensely by Dolops carvalhoi, in situation of high population density. Piaractus mesopotamicus was considered the species more parasited by myxosporídeos, that were also observed in L. macrocephalus and cross-breeding tambacu.