Tadpoles fed supplemented diet with probiotic bacterium isolated from the intestinal tract of bullfrog Lithobates catesbeianus: Haematology, cell activity and electron microscopy.

The aim of this study is to select and isolate autochthonous bacteria with probiotic potential for use in a supplemented diet for bullfrog tadpoles, Lithobates catesbeianus. A total of 20 strains of lactic acid bacteria were isolated. Nine out of these were used in the following in vitro assays: antagonism against pathogenic bacteria (ANT), antimicrobial activity from extracellular compounds (MIC), tolerance to bile salts (TBS), pH reduction, protease production, sensitivity to antimicrobial tetracycline, cell viability, growth rate and doubling time. Using these data was defined an ideotype (ideal strain) based on the best results. Distances were estimated with the Mahalanobis (D2) test, and the best candidates, presenting the shortest ideotype distances, were considered to be used. The best strain was found to be Lactobacillus plantarum because it presented 10.00 ± 0.50 mm of ANT against Aeromonas hydrophila, 3.99 ± 0.01 of MIC independent of pathogenic bacteria, 85.07 ± 0.01 of TBS, 4.20 ± 0.02 of final pH, 17.67 ± 1.15 of protease production, 13.50 ± 2.00 sensitivity to antimicrobial tetracycline, 9.36 ± 0.04 of cell viability, 0.20 ± 0.00 of growth rate and 3.46 ± 0.00 doubling time. Therefore this probiotic candidate was then supplemented (2.045 ± 1.07 × 107 colony forming unities. g-1) into the diets of bullfrog tadpoles for a period of 42 days. At the end of the trial, samples of blood and intestines were collected to verify the haematological alterations and the intestinal morphology using transmission and scanning electron microscopy. Tadpoles fed the supplemented diet showed successful lactic acid bacterium colonisation, an increased number of circulating thrombocytes, monocytes, eosinophil and LG-PAS+ and also an increase in the length and density of intestinal microvilli. This study shows the feasibility of using probiotics isolated from farmed bullfrogs as a supplement in the diets of tadpoles, providing a promising alternative for modulating the health of these animals.

Effect of water temperature and salinity in oviposition, hatching success and infestation of Aphanoblastella mastigatus (Monogenea, Dactylogyridae) on Rhamdia quelen.

Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasite's survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatus's eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Effect of water temperature and salinity in oviposition, hatching success and infestation of Aphanoblastella mastigatus (Monogenea, Dactylogyridae) on Rhamdia quelen / Efeito da temperatura da água e salinidade na oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus (Monogenea, Dactylogyridae) em Rhamdia quelen

Abstract Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasite’s survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatus’s eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Resumo Diversos parâmetros ambientais podem afetar os processos biológicos de diversos organismos invertebrados aquáticos, como os Monogenea. Neste estudo, nós investigamos a oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus, parasito de jundiá Rhamdia quelen, em diferentes condições de temperatura (~ 24 e 28 °C) e salinidade (por meio da adição de cloreto de sódio na água nas concentrações de 0, 5 e 9 g/L) em laboratório. Não houve diferença significativa na taxa de oviposição nem no sucesso de infestação de A. mastigatus à 24 e 28 °C; por outro lado, a concentração de 9 g/L de cloreto de sódio na água inviabilizou não só a sobrevivência do parasito mas também a viabilidade dos ovos. Esse resultado indica que seu uso é eficiente como potencial tratamento profilático. A taxa de eclosão dos ovos de A. mastigatus foi significativamente maior a 28 °C. A partir de 5 g/L, a mesma foi significativamente menor. Durante um período de 24 horas, dois picos na oviposição ocorreram nos momentos de maior variação da luminosidade (06h15 e 18h15). Futuros estudos são recomendados utilizando intervalos de temperatura e intensidades de infestação experimental maiores para verificar a influência da temperatura na longevidade e sucesso de infestação de A. mastigatus.

Effect of water temperature and salinity in oviposition, hatching success and infestation of Aphanoblastella mastigatus (Monogenea, Dactylogyridae) on Rhamdia quelen

Abstract Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasites survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatuss eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Resumo Diversos parâmetros ambientais podem afetar os processos biológicos de diversos organismos invertebrados aquáticos, como os Monogenea. Neste estudo, nós investigamos a oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus, parasito de jundiá Rhamdia quelen, em diferentes condições de temperatura (~ 24 e 28 °C) e salinidade (por meio da adição de cloreto de sódio na água nas concentrações de 0, 5 e 9 g/L) em laboratório. Não houve diferença significativa na taxa de oviposição nem no sucesso de infestação de A. mastigatus à 24 e 28 °C; por outro lado, a concentração de 9 g/L de cloreto de sódio na água inviabilizou não só a sobrevivência do parasito mas também a viabilidade dos ovos. Esse resultado indica que seu uso é eficiente como potencial tratamento profilático. A taxa de eclosão dos ovos de A. mastigatus foi significativamente maior a 28 °C. A partir de 5 g/L, a mesma foi significativamente menor. Durante um período de 24 horas, dois picos na oviposição ocorreram nos momentos de maior variação da luminosidade (06h15 e 18h15). Futuros estudos são recomendados utilizando intervalos de temperatura e intensidades de infestação experimental maiores para verificar a influência da temperatura na longevidade e sucesso de infestação de A. mastigatus.

New species of Cucullanus (Nematoda: Cucullanidae), an intestinal parasite of the peacock bass Cichla piquiti (Perciformes: Cichlidae) from the Tocantins River, Brazil.

Cucullanus tucunarensis n. sp. (Cucullanidae) is described parasitizing the peacock bass Cichla piquiti (Cichlidae) from the Tocantins River, Tocantins State, Brazil. The new species is unique and differs from all its congeners by having a tail tip provided with several sclerotized spine-like processes on its ventral side, present in both the male and female. Furthermore, C. tucunarensis n. sp. is compared with other species of the genus recorded in the same zoogeographical region and in the same group of hosts (Perciformes).

Helicometrina nimia Linton, 1910 (Digenea: Opecoelidae) in dusky grouper Epinephelus marginatus (Lowe, 1834) (Teleostei: Serranidae) from southeastern Brazil.

Helicometrina nimia (Opecoelidae) is a digenean with wide distribution. Fish families most commonly used as hosts for H. nimia are Serranidae, Pomodasydae, Scorpaenidae and Clinidae. In the present study, a new host and a new host locality are presented for the species. A description of the studied specimens, besides comments concerning its taxonomic status and biometrically compared tables of H. nimia reports are given. The taxonomic status of members of Helicometrina has been questionable. The greatest controversy for the genus seems to be related to the validity of diagnostic features, especially in regard to the number of testes. In the present study, all studied specimens presented a permanent and steady number of testes (n=9) and therefore its use as a diagnostic character is supported by the present authors. Epinephelus marginatus is considered a new host for Helicometrina nimia, and São Paulo state, southeastern Brazil, a new locality for the species.

Helicometrina nimia Linton, 1910 (Digenea: Opecoelidae) in dusky grouper Epinephelus marginatus (Lowe, 1834) (Teleostei: Serranidae) from southeastern Brazil / Helicometrina nimia Linton, 1910 (Digenea: Opecoelidae) em garoupa verdadeira Epinephelus marginatus (Lowe, 1834) (Teleostei: Serranidae) no Sudeste do Brasil

Helicometrina nimia (Opecoelidae) is a digenean with wide distribution. Fish families most commonly used as hosts for H. nimia are Serranidae, Pomodasydae, Scorpaenidae and Clinidae. In the present study, a new host and a new host locality are presented for the species. A description of the studied specimens, besides comments concerning its taxonomic status and biometrically compared tables of H. nimia reports are given. The taxonomic status of members of Helicometrina has been questionable. The greatest controversy for the genus seems to be related to the validity of diagnostic features, especially in regard to the number of testes. In the present study, all studied specimens presented a permanent and steady number of testes (n=9) and therefore its use as a diagnostic character is supported by the present authors. Epinephelus marginatus is considered a new host for Helicometrina nimia, and São Paulo state, southeastern Brazil, a new locality for the species.

Helicometrina nimia (Opecoelidae) é um digenético com distribuição mundial. As famílias de peixes mais comumente utilizadas como hospedeiros por H. nimia são Serranidae, Pomodasydae, Scorpaenidae e Clinidae. No presente estudo, um novo hospedeiro e uma nova localidade são apresentados para a espécie. A descrição dos espécimes estudados, além de comentários sobre o seu status taxonômico e uma tabela comparativa de medidas de H. nimia são apresentados. O status taxonômico dos membros de Helicometrina tem sido questionável. A maior controvérsia dentro do gênero parece estar relacionada com a validade das características diagnósticas, especialmente no que diz respeito ao número de testículos. No presente estudo, todos os espécimes estudados apresentaram número permanente e estável de testículos (n=9) e, portanto, sua utilização como caráter diagnóstico é suportada pelos presentes autores. Epinephelus marginatus é considerada um novo hospedeiro para Helicometrina nimia e São Paulo, sudeste do Brasil, uma nova localidade para a espécie.

Parasite fauna of wild and cultured dusky-grouper Epinephelus marginatus (Lowe, 1834) from Ubatuba, southeastern Brazil / Fauna parasitária da garoupa selvagem e de cultivo Epinephelus marginatus (Lowe, 1834) em Ubatuba, sudeste do Brasil

This study aimed at identifying and quantifying the parasites of wild and cultured dusky grouper Epinephelus marginatus. During a year and thereby all four seasons, 20 wild and 20 cultured groupers were examined for the presence of parasites, except in the last season, in which 19 wild and 20 cultured fish were examined, totalling 159 groupers analysed from Ubatuba, southeastern Brazil. Prevalence, mean intensity of infection, mean abundance and mean relative dominance were calculated. Five species of parasites were identified in fish from both origins: Pseudorhabdosynochus beverleyburtonae (Monogenea), Neobenedenia melleni (Monogenea), Pseudempleurosoma sp. (Monogenea), Helicometrina nimia (Digenea) and larvae of Contracaecum sp. (Nematoda). The prevalence of ectoparasites, in most cases, was higher than endoparasites. The most abundant parasite was the monogenea Pseudorhabdosynochus beverleyburtonae in both wild and cultured fish, along all seasons. Neobenedenia melleni was observed in wild and cultured fish in all seasons, with a gradual increase in the number of parasites from the coldest to the hottest seasons, with the highest prevalence and mean intensity in the summer. Helicometrina nimia was found in all seasons in both wild and cultured fish, except for summer, where its presence was detected only in wild fish. Pseudempleurosoma sp. and larvae of Contracaecum sp. showed low prevalence occurring in wild and cultured fish in the autumn and spring, respectively. This study revealed high intensities of potentially pathogenic parasites that could favour disease outbreaks in culture conditions.

Este estudo teve como objetivo identificar e quantificar os parasitos da garoupa verdadeira Epinephelus marginatus selvagem e cultivada. Durante um ano, nas quatro estações, 20 garoupas selvagens e 20 cultivadas foram examinadas quanto à presença de parasitos, exceto na última estação, em que foram examinados 19 peixes selvagens e 20 cultivados, totalizando 159 peixes examinados em Ubatuba, Sudeste do Brasil. A prevalência, intensidade média de infecção, abundância média and dominância média relativa foram calculados. Cinco espécies de parasitos foram identificadas nos peixes de ambas as origens: Pseudorhabdosynochus beverleyburtonae (Monogenea), Neobenedenia melleni (Monogenea), Pseudempleurosoma sp. (Monogenea), Helicometrina nimia (Digenea) e larvas de Contracaecum sp. (Nematoda). A prevalência de ectoparasitos foi, na maioria das vezes, maior do que a de endoparasitos. O parasito mais abundante foi o monogenea Pseudorhabdosynochus beverleyburtonae em ambos os peixes selvagens e cultivados, durante todas as estações. Neobenedenia melleni foi observado e, peixes selvagens e cultivados em todas as estações, com aumento gradativo no número de parasitos das estações mais frias para as mais quentes, com as maiores prevalências e intensidades médias no verão. Helicometrina nimia foi encontrado em todas as estações em ambos os peixes selvagens e cultivados, exceto no verão, em que foi observada sua presença apenas em peixes selvagens. Pseudempleurosoma sp. e larvas de Contracaecum sp. apresentaram baixas prevalências, ocorrendo em peixes selvagens e cultivados no outono e na primavera, respectivamente. Este estudo revelou altas intensidades de parasitos potencialmente patogênicos que podem favorecer surtos de doenças em situações de cultivo.

Parasite fauna of wild and cultured dusky-grouper Epinephelus marginatus (Lowe, 1834) from Ubatuba, southeastern Brazil.

This study aimed at identifying and quantifying the parasites of wild and cultured dusky grouper Epinephelus marginatus. During a year and thereby all four seasons, 20 wild and 20 cultured groupers were examined for the presence of parasites, except in the last season, in which 19 wild and 20 cultured fish were examined, totalling 159 groupers analysed from Ubatuba, southeastern Brazil. Prevalence, mean intensity of infection, mean abundance and mean relative dominance were calculated. Five species of parasites were identified in fish from both origins: Pseudorhabdosynochus beverleyburtonae (Monogenea), Neobenedenia melleni (Monogenea), Pseudempleurosoma sp. (Monogenea), Helicometrina nimia (Digenea) and larvae of Contracaecum sp. (Nematoda). The prevalence of ectoparasites, in most cases, was higher than endoparasites. The most abundant parasite was the monogenea Pseudorhabdosynochus beverleyburtonae in both wild and cultured fish, along all seasons. Neobenedenia melleni was observed in wild and cultured fish in all seasons, with a gradual increase in the number of parasites from the coldest to the hottest seasons, with the highest prevalence and mean intensity in the summer. Helicometrina nimia was found in all seasons in both wild and cultured fish, except for summer, where its presence was detected only in wild fish. Pseudempleurosoma sp. and larvae of Contracaecum sp. showed low prevalence occurring in wild and cultured fish in the autumn and spring, respectively. This study revealed high intensities of potentially pathogenic parasites that could favour disease outbreaks in culture conditions.

Morphology of larval Gordius dimorphus (Nematomorpha: Gordiida).

Scanning electron microscopy was used to analyze superficial features of Gordius dimorphus Poinar, 1991, larvae that might serve as generic or specific diagnostic characters. Three adults of G. dimorphus (2 males and 1 female) were maintained under laboratory conditions until oviposition, which occurred within long strings commonly referred to as egg strings. Larvae have a cylindrical body, annulated and divided into 2 sections, plus an anterior preseptum and a posterior postseptum. Three concentric rings with 6 spines each surround the proboscis. The proboscis is retractile, dorsoventrally flattened, with 1 pair of forceps-like projections on its distal-most portion. On the surface of each projection, 3 pairs of aculeiforms spines are aligned and lean toward the proboscis opening. One large spine is present on the posterior portion of the postseptum. Papillae were not observed. Gordius dimorphus larvae are similar to previous light microscope descriptions. This is the first record of G. dimorphus in Brazil.