Geographic range, distribution patterns and interactions of Monogenea Van Beneden 1858, with species of native host freshwater fishes from Brazil / Distribuição geográfica, padrões de distribuição e interações de Monogenea Van Beneden 1858, com espécies de peixes hospedeiros nativos de água doce do Brasil

Abstract This paper investigated information on monogenean species using 312 scientific papers, to search for infection and geographic distribution patterns in native freshwater fish from Brazil. We used 1,698 samples of 296 fish species of 28 families distributed into Characiformes, Siluriformes, Cichliformes, Gymnotiformes, Perciformes, Mugiliformes, Osteoglossiformes and Clupeiformes, in addition to four hybrid fish. Among the hosts of the different orders and families, the greatest numbers of parasite-host associations were found for species of the families Serrasalmidae, Characidae, Loricariidae, Curimatidae and Anostomidae. The 578 species of monogeneans used in parasite-host interactions were distributed in 86 genera of six five families (Dactylogyridae, Gyrodactylidae, Diplectanidae, Microcotylidae, Ancylodiscoididae and Ancyrocephalidae), but with great predominance of Dactylogyridae species. There was variation in prevalence, intensity and abundance levels of monogeneans species among host fish species, as well as in infection sites that occurred predominantly in external organs. Positive correlations of prevalence, intensity and abundance with body length of hosts were observed. There was geographic distribution pattern of monogeneans limited mostly to two hydrographic basins those being the Amazon River and Paraná River. Just approximately 6% of potential monogeneans have been explored thus far, showing a clear need for further studies on this interesting group of parasites.

Resumo Este estudo investigou informações sobre espécies de monogenéticos, usando 312 artigos científicos para buscar padrões de infecção e distribuição geográfica em peixes nativos de água doce do Brasil. Foram utilizadas 1.698 amostras de 296 espécies de peixes de 28 famílias, distribuídas em Characiformes, Siluriformes, Cichliformes, Gymnotiformes, Perciformes, Mugiliformes, Osteoglossiformes e Clupeiformes, além de quatro peixes híbridos. Entre os hospedeiros das diferentes ordens e famílias, os maiores números de associações parasito-hospedeiro foram encontrados para espécies das famílias Serrasalmidae, Characidae, Loricariidae, Curimatidae e Anostomidae. As 578 espécies de monogenéticos, utilizadas nas interações parasito-hospedeiro, foram distribuídas em 86 gêneros de seis famílias (Dactylogyridae, Gyrodactylidae, Diplectanidae, Microcotylidae, Ancylodiscoididae e Ancyrocephalidae), mas com grande predominância de espécie de Dactylogyridae. Houve variação nos níveis de prevalência, intensidade e abundância das espécies de monogenéticos entre as espécies de peixes hospedeiros, bem como nos locais de infecção que ocorreram predominantemente em órgãos externos. Correlações positivas de prevalência, intensidade e abundância com o comprimento corporal dos hospedeiros foram observadas. Houve padrão de distribuição geográfica dos monogenéticos limitados principalmente a duas bacias hidrográficas, sendo elas o Rio Amazonas e Rio Paraná. Apenas aproximadamente 6% dos potenciais monogenéticos são conhecidos até agora, mostrando uma clara necessidade de mais estudos sobre esse interessante grupo de parasitos.

Snail transmitters of schistosomiasis and other mollusks of medical and economic importance at the simplício queda única hydroelectric plant, southeast Brazil

Abstract INTRODUCTION This paper presents the results of an extensive survey of freshwater mollusks in the Simplício Queda Única Hydroelectric Development area, Southeast Brazil. METHODS Mollusks were collected between 2008 and 2013. All specimens were examined for the presence of larval trematodes. RESULTS In total, 12,507 specimens classified into 16 genera were obtained. Known snail vectors of schistosomiasis and fascioliasis and exotic species were identified, and new records are reported. CONCLUSIONS No specimens parasitized by larval trematodes of medical interest were detected. However, the results reinforce the importance of surveillance in study areas vulnerable to the occurrence of schistosomiasis transmission.

Infectivity of Chordodes nobilii larvae (Gordiida: Nematomorpha) / Infectividad de las larvas de Chordodes nobilii (Gordiida: Nematomorpha)

AbstractThe gordiids are freshwater representatives of the parasite phylum Nematomorpha that function as a link between aquatic and terrestrial ecosystems. In recent years, different ecotoxicologic studies have been made with the South-American gordiid species, Chordodes nobilii, that have demonstrated the capacity of this group to act as a bioindicator of contamination. Despite the Gordiida's ecologic relevance, further studies are still needed to elucidate different aspects of the biology of the class, and among those, the infective capacity, a parameter that can be evaluated by the infection index mean abundance (IIMA). A knowledge of the intrinsic variability in the infective capacity of C. nobilii would warrant priority in order to establish, the range of acceptable responses for normal or standard conditions in the laboratory, and, to compare the criteria among different assays. The objective of this study was to establish a baseline value for the infective capacity for C. nobilii larvae, under controlled laboratory conditions, by employing the IIMA as the evaluation parameter. To that end, we analyzed the infective capacity of C. nobilii larvae that had hatched from different strings of eggs laid in the laboratory by a total of 12 females. The C. nobilii adults were collected from streams within the Argentina Sauce Grande basin, between 2006 and 2009. Once in the laboratory, after mating, the females were placed in individual containers for oviposition. The egg strings obtained from each female were cut in 3 mm long segments; and when free larvae were observed, the segments (N= 90) were placed together with 30 Aedes aegypti larvae for evaluation of the gordiids' infective capacity. After 72 h, the mosquito larvae were observed by microscopy in order to quantify the C. nobilii larvae in body cavities. The IIMAs were calculated as the total number of C. nobilii larvae present divided by total number of A. aegypti larvae examined. For analysis of the IIMAs obtained, the data were grouped according to the female who made the original ovoposition. Our results enabled the corroboration of an ample range of responses in the infective capacity of this species, a characteristic that would normally be linked to the progenitors originating the hatch. Because this relationship prevents the establishment of a baseline for making comparisons among assays with gordiids, through the IIMA as a response parameter, we recommend expressing the IIMA values in each assay relative to their respective controls. These findings also provide evidence for the greater success in infections by certain members of the progeny over others. Finally, on the basis of the results obtained from this study, we stress the relevance of the use of the IIMA as a decisive aspect to be considered in different studies on the biology of Gordiida. Rev. Biol. Trop. 65 (1): 1-8. Epub 2017 March 01.

ResumenLos gordiidos son representantes dulceacuícolas del Phylum parásito Nematomorpha que actúan como un enlace entre ecosistemas acuáticos y terrestres. En años recientes, diferentes estudios ecotoxicológicos se han desarrollado con una especie sudamericana de gordiido, C. nobilii, que ha demostrado la capacidad de este grupo de actuar como bioindicador de contaminación. A pesar de su evidente importancia ecológica, aún se necesitan realizar estudios para dilucidar distintos aspectos de su biología, entre estos, la capacidad infectiva, un parámetro que puede evaluarse utilizando el Índice de Infección Abundancia Media (IIMA). El conocimiento de la variabilidad intrínseca en la capacidad infectiva de C. nobilii merece prioridad con el objeto de establecer el ámbito de respuesta aceptable para condiciones normales o estándar en el laboratorio, y que permita comparar los resultados entre distintos ensayos. El objetivo de este estudio es establecer la línea de base de la capacidad infectiva del gordiido C. nobilii en condiciones controladas de laboratorio, empleando el IIMA como parámetro de evaluación. Con este fin, se analizó la capacidad infectiva de larvas de C. nobilii que eclosionaron de diferentes cordones de huevos depositados por un total de 12 hembras, mantenidas en laboratorio. Los adultos de C. nobilii fueron recolectados de arroyos de la cuenca argentina Sauce Grande, entre 2006 y 2009. Una vez en el laboratorio, después de la cópula, las hembras se ubicaron en recipientes individuales a la espera de la oviposición. Los cordones de huevos obtenidos de cada hembra se cortaron en segmentos de 3 mm de longitud; y cuando las larvas libres fueron observadas al microscopio, los segmentos (N= 90) fueron ubicados junto con 30 larvas de Aedes aegypti para evaluar la capacidad infectiva del gordiido. Después de 72 h, las larvas del mosquito fueron observadas al microscopio para contabilizar las larvas de C. nobilii en las cavidades corporales. El IIMA fue calculado como el número total de larvas de C. nobilii presentes dividido entre el número total de larvas de A. aegypti examinadas. Para el análisis de los IIMAs obtenidos, los datos fueron agrupados de acuerdo a la hembra que hizo la oviposición. Nuestros resultados permiten corroborar un amplio rango de respuesta en la capacidad infectiva de esta especie, que estaría vinculada al origen de la camada. Debido a que no se pudo establecer una línea de base para realizar comparaciones entre estudios en los gordiidos utilizando el IIMA como parámetro de respuesta, se aconseja relativizar los valores de los IIMAs a sus respectivos controles. Estos resultados también pusieron en evidencia la ventaja en el éxito de infección de algunas progenies sobre las restantes. Finalmente, con base en los resultados obtenidos a partir de este estudio se plantea la importancia del uso del IIMA como punto final a considerar en distintos estudios sobre la biología de los gordiida.

Waterborne toxoplasmosis investigated and analysed under hydrogeological assessment: new data and perspectives for further research

We present a set of data on human and chicken Toxoplasma gondiiseroprevalence that was investigated and analysed in light of groundwater vulnerability information in an area endemic for waterborne toxoplasmosis in Brazil. Hydrogeological assessment was undertaken to select sites for water collection from wells for T. gondiioocyst testing and for collecting blood from free-range chickens and humans for anti-T. gondiiserologic testing. Serologic testing of human specimens was done using conventional commercial tests and a sporozoite-specific embryogenesis-related protein (TgERP), which is able to differentiate whether infection resulted from tissue cysts or oocysts. Water specimens were negative for the presence of viable T. gondiioocysts. However, seroprevalence in free-range chickens was significantly associated with vulnerability of groundwater to surface contamination (p < 0.0001; odds ratio: 4.73, 95% confidence interval: 2.18-10.2). Surprisingly, a high prevalence of antibodies against TgERP was detected in human specimens, suggesting the possibility of a continuous contamination of drinking water with T. gondiioocysts in this endemic setting. These findings and the new proposed approach to investigate and analyse endemic toxoplasmosis in light of groundwater vulnerability information associated with prevalence in humans estimated by oocyst antigens recognition have implications for the potential role of hydrogeological assessment in researching waterborne toxoplasmosis at a global scale.

The hyporheic zone and its functions: revision and research status in Neotropical regions / A zona hiporréica e as suas funções: revisão e estado da arte da pesquisa na região neotropical

AbstractThe hyporheic zone (HZ), as the connecting ecotone between surface- and groundwater, is functionally part of both fluvial and groundwater ecosystems. Its hydrological, chemical, biological and metabolic features are specific of this zone, not belonging truly neither to surface- nor to groundwater. Exchanges of water, nutrients, and organic matter occur in response to variations in discharge and bed topography and porosity. Dynamic gradients exist at all scales and vary temporally. Across all scales, the functional significance of the HZ relates to its activity and connection with the surface stream. The HZ is a relatively rich environment and almost all invertebrate groups have colonized this habitat. This fauna, so-called hyporheos, is composed of species typical from interstitial environment, and also of benthic epigean and phreatic species. The hyporheic microbiocenose consists in bacteria, archaea, protozoa and fungi. The HZ provides several ecosystem services, playing a pivotal role in mediating exchange processes, including both matter and energy, between surface and subterranean ecosystems, functioning as regulator of water flow, benthic invertebrates refuge and place of storage, source and transformation of organic matter. The hyporheic zone is one of the most threatened aquatic environments, being strongly influenced by human activities, and the least protected by legislation worldwide. Its maintenance and conservation is compelling in order to preserve the ecological interconnectivity among the three spatial dimensions of the aquatic environment. Although several researchers addressed the importance of the hyporheic zone early, and most contemporary stream ecosystem models explicitly include it, very little is known about the HZ of Neotropical regions. From a biological standpoint, hyporheos fauna in Neotropical regions are still largely underestimated. This review focuses on a brief presentation of the hyporheic zone and its functions and significance as an ecotone. We also highlighted the key aspects considering also the current status of research in Neotropical regions.

ResumoA zona hiporréica, como ecótono de ligação entre a superfície e as águas subterrâneas, é parte funcional seja dos ecossistemas fluviais seja das águas subterrâneas. As características hidrológicas, as características químicas, biológicas e metabólicas são específicas desta zona, não pertencendo verdadeiramente nem a superfície nem às águas subterrâneas. Trocas de água, nutrientes e matéria orgânica ocorrem em resposta a variações na descarga, topografia do álveo e porosidade. Gradientes dinâmicos existem em todas as escalas e variam temporalmente. Em todas as escalas, o significado funcional da zona hyporheic relaciona-se com a sua conexão e atividades com a água superficial. O HZ é um ambiente relativamente rico e quase todos os grupos de invertebrados colonizaram este habitat. Esta fauna, chamada hyporheos, é composta por espécies típicas do ambiente intersticial, e também de espécies bentônicas epígeas e freáticas. A microbiocenose consiste em bactérias, arqueobactérias, fungos e protozoários. O HZ fornece vários serviços para o ecossistema, desempenhando um papel fundamental na mediação de processos de troca, incluindo seja a matéria, seja a energia, entre os ecossistemas superfíciais e os subterrâneos, funcionando como regulador do fluxo de água, de refúgio para invertebrados bentônicos e local de armazenagem, fonte e transformação de matéria orgânica. A zona hyporheic é um dos ambientes aquáticos mais ameaçados, sendo fortemente influenciado pelas atividades humanas, e um dos menos protegidos pela legislação em todo o mundo. A sua manutenção e conservação é necessaria para preservar a interconectividade ecológica entre as três dimensões espaciais do ambiente aquático. Apesar de vários pesquisadores aborem a importância da zona hyporheic a tempo, e a maioria dos modelos de ecossistemas atualmente incluí-lo de forma explicita, muito pouco se sabe sobre o HZ das regiões neotropicais. Do ponto de vista biológico, a fauna hiporréica das regiões neotropicais é ainda largamente subestimada. Esta revisão visa apresentar de forma resumida a zona hiporréica, suas funções e importância como ecótono. Também visa destacar os aspectos principais considerando também o estado actual da investigação em regiões neotropicais.

Trematode Metacercariae in Freshwater Fish from Water Systems of Hantangang and Imjingang in Republic of Korea

The infection status of freshwater fish with digenetic trematode metacercariae was examined in water systems of Hantangang and Imjingang (River), the Republic of Korea. A total of 877 (594 from Hantangang and 283 from Imjingang) fishes were examined by the artificial digestion methods. Clonorchis sinensis metacercariae were detected in 7 (1.2%) fishes (in 3 spp.) from Hantangang in Cheorwon-gun, Gangwon-do, and 40 (14.1%) fishes (in 7 spp.) from Munsancheon in Paju-si, Gyeonggi-do. The average densities were 1.9 and 35.6 per fish infected, respectively. Metagonimus spp. metacercariae were detected in 312 (52.5%) and 113 (39.9%) fishes from Hantangang and Imjingang, and their average densities were 47.5 and 9.6 per fish infected, respectively. Centrocestus armatus metacercariae were found in 161 (27.1%) and 70 (24.7%) fishes from Hantangang and Imjingang, and their average number per fish infected was 694 and 82, respectively. Echinostoma spp. metacercariae were detected in 50 (8.4%) and 94 (33.2%) fishes from Hantangang and Imjingang, and their average densities were 9.6 and 23.1 per fish infected, respectively. The infection status of fishes with metacercariae of Stephanoprora spp., Diplostomum spp., Clinostomum complanatum, Metorchis orientalis, and Metorchis taiwanensis were analyzed by surveyed regions. Conclusively, it was confirmed that C. sinensis metacercariae were quite commonly detected in fishes from Munsancheon but rarely from other localities, whereas the metacercariae of other digenetic trematodes were relatively prevalent in fishes from water systems of Hantangang and Imjingang in Korea.

The importance of protozoan bacterivory in a subtropical environment (Lobo-Broa Reservoir, SP, Brazil) / A importância da bacterivoria por protozoários em um ambiente subtropical (Reservatório do Lobo-Broa, SP, Brasil)

This study evaluated the importance of heterotrophic nanoflagellates (HNF) and ciliates bacterivory in a mesotrophic subtropical environment (Lobo-Broa Reservoir, Brazil) by the quantification of their ingestion rates. The in situ experiments using fluorescently labelled bacteria (FLB) were carried out bimonthly over one year (three surveys in the dry season and three in the rainy one) at the sub-surface of two sampling points that have different trophic degrees. The ingestion rates for both ciliates and HNF were higher in the meso-eutrophic region (point 2) due to the higher water temperatures, which accelerate the metabolism of protozoans and the higher bacteria densities. Concerning total protozoan bacterivory, the HNF had the greatest grazing impact on bacterial community, especially the HNF <5µm. The data showed that HNF grazing, in addition to regulating the bacteria abundance, also induced changes to the bacterial community structure, such as increasing size and numbers of bacterial filaments. The ciliates were also important to the system bacterivory, especially in point 2, where there were high densities and ingestion rates. The protozoan bottom-up control was more important in the dry season and the top-down control was more important in the rainy season, so, these two forces are equally important to the bacterial abundance regulation in this reservoir in an annual basis.

Este estudo avaliou a importância da bacterivoria por nanoflagelados heterotróficos (NFH) e ciliados em um ambiente subtropical mesotrófico (Reservatório do Lobo-Broa, Brasil) pela quantificação de suas taxas de ingestão. Os experimentos in situ utilizando bactérias marcadas por fluorescência (FLB) foram realizados bimestralmente durante um ano (três estudos no período seco e três no período chuvoso) na superfície de dois pontos de coleta que apresentam graus de trofia diferentes. As taxas de ingestão, tanto para os ciliados quanto para os NHF, foram maiores no ponto 2 (P2) devido provavelmente as maiores temperaturas da água, que aceleram o metabolismo dos protozoários, e as maiores densidades de bactérias. Em relação à bacterivoria total por protozoários, as populações de NFH causaram maior impacto de predação sobre a comunidade bacteriana do reservatório, principalmente os NFH menores que 5µm. Os dados indicaram que a predação por NFH, além de regular a abundância de bactérias, provavelmente também induziu modificações na estrutura da comunidade bacteriana, como o aumento do tamanho e da quantidade de filamentos bacterianos. Os ciliados também contribuíram significativamente para a bacterivoria no sistema, principalmente no P2, onde apresentaram altas densidades e taxas de ingestão. O controle bottom-up teve maior importância no período seco e o controle top-down maior importância no período chuvoso, assim, estas duas forças são igualmente importantes na regulação da abundância bacteriana neste reservatório em uma base anual.

A malacological survey in the Manso Power Plant, State of Mato Grosso, Brazil: new records of freshwater snails, including transmitters of schistosomiasis and exotic species

Introduction Schistosomiasis is a parasitic disease of public health concern in Brazil, and the construction of hydroelectric dams, in addition to increasing permanent human settlement and tourism, has created conditions suitable for the establishment of mollusks that can transmit schistosomiasis. Such areas require a number of actions to prevent the establishment of schistosomiasis. This paper reports on a freshwater malacological survey carried out in the geographical area of the Manso Power Plant. Methods Mollusks were collected in 18 municipalities in the State of Mato Grosso between February 2002 and February 2004 (qualitative study) and from April 2009 to February 2011 (quantitative study). Results Thirty-one species of mollusks were collected, including newly recorded species (Antillorbis nordestensis and Burnupia ingae). In addition, the geographic distributions of known species, including Biomphalaria straminea, a snail vector of Schistosoma mansoni, were expanded. A total of 4,507 specimens were collected in the APM Manso reservoir (Usina Hidrelétrica de Aproveitamento Múltiplo de Manso) during the quantitative study, and Biomphalaria amazonica was found in six of the 10 localities analyzed. The Afroasiatic species Melanoides tuberculata, introduced after February 2009, was the dominant species (relative abundance 94.96%). Conclusions The study area is epidemiologically important due to the occurrence of B. straminea and B. amazonica, which are vectors of schistosomiasis, and M. tuberculata, a snail host of Centrocestus formosanus, which is responsible for centrocestiasis transmission. Observations of M. tuberculata and the exotic freshwater clams Corbicula fluminea and Corbicula largillierti raise concerns about biodiversity. .

Pfeifferinella SP. [pfeifferinellidae, apicomplexan] infecting the fresh water snail pirenella conica light and electron microscope studies

Coccidian oocysts were proved to be found in 70 of 100 collected Pirenella conica snails, with a natural infection of 70%. It was observed that, Pfeifferinella sp. was transferred between hepatopancreas and small intestine of snail. The prepatent period of Pfeifferinella sp. infecting P. conica snails ranged from 14-18 days and the patent period was reached 50 days [P.L]- Mer-ogony stages were the early stages observed in this study. These stages were observed in the hepatopancreas and in a large clear parasiteophorous vacuole [PV]. In snails killed 4 days P.I. immature meronts were measured 12x10 microm containing 8 nuclei. Meanwhile, mature meronts with about 6 differentiated merozoites were detected as early as 6 days P.I., and measured 3.1xl.4jim. The earliest gametogonic stages were seen in the intestine of Pirenella conica snails killed 12 days P.L Microgamonts contained about 4 nuclei and measured 79x6.7microm. The mac-rogamonts measured 7.3x5.6|

Melena revealing a nasopharyngeal leech: a paediatric case

Leech infestation occurs after swimming or ingestion of freshwater, especially in rural areas. Symptoms are different depending on the binding site of the leech. At the nasopharynx, it is mainly responsible for epistaxis. We report a case of an unknown pharyngeal location of a leech in a three year-old girl responsible for melena and leading to a severe anaemia

Polymerase chain reaction and nested-PCR approaches for detecting Cryptosporidium in water catchments of water treatment plants in Curitiba, State of Paraná, Brazil

Introduction Cryptosporidium is an important protozoan cause of waterborne disease worldwide of concern to public health authorities. To prevent outbreaks of cryptosporidiosis, the monitoring of this parasite in drinking water is necessary. In the present work, the polymerase chain reaction (PCR) and nested-PCR techniques were used to detect Cryptosporidium in raw water from catchment points of four water treatment plants (WTP) in Curitiba, Paraná, Brazil. Methods First, DNA extraction techniques were tested in samples containing decreasing amount of oocysts in reagent water, and PCR and nested-PCR with specific primers for 18SSU rDNA of Cryptosporidium were conducted to determine their sensitivity. In reagent water, a commercial extraction kit provided the best analytical sensitivity, and PCR and nested-PCR allowed the detection of five and two oocysts, respectively, with the primers XIAOR/XIAOF and XIAO1F/XIAO2R. Results In the spiking experiments, only the PCR with the primers AWA995F/AWA1206R was successful at detecting concentrations of 0.1 oocysts/mL. Two catchments samples of raw water and/or water sludge from four WTPs were contaminated with Cryptosporidium. Conclusions The application of the techniques to monitor Cryptosporidium in water and detect contamination in water catchments of WTPs in Curitiba are discussed in the present work. .

Surtos de infecção por Piscinoodinium pillulare e Henneguya spp. em pacus (Piaractus mesopotamicus) criados intensivamente no Sudoeste de Goiás / Outbreaks of Piscinoodinium pillulare and Henneguya spp. infection in intensively raised Piaractus mesopotamicus in Southwestern Goiás, Brazil

Três surtos de infecção por Piscinoodinium pillulare e Henneguya spp. em Piaractus mesopotamicus criados intensivamente no Sudoeste de Goiás são descritos. Dois surtos ocorreram no inverno e um no verão. As taxas de morbidade (58,57%-90%) e mortalidade (100%) foram determinadas. Episódios de estresse ocorreram previamente em todos os casos. Sinais clínicos incluíam mudanças comportamentais e movimentos erráticos com perda de equilíbrio. Os principais achados macroscópicos foram tumefação das brânquias com exsudato mucoso esbranquiçado. Microscopicamente, as brânquias apresentaram inflamação linfoplasmocítica e necrosante, multifocal, leve a moderada, com hipertrofia e fusão de lamelas secundárias. Adicionalmente, numerosos trofontes intralesionais de P. pillulare e cistos de Henneguya spp. foram notados. Para o conhecimento dos autores, essa parece ser a primeira descrição de infecção por P. pillulare e Henneguya spp. em peixes criados no Sudoeste de Goiás.

Three outbreaks of infection by Piscinoodinium pillulare and Henneguya spp. in Piarac tus mesopotamicus intensively raised in Southwestern Goiás, Brazil, are described. Two outbreaks occurred in the winter and another one in summer. Morbidity (58.57%-90%) and mortality (100%) rates were determined. Stress episodes occurred previously in all cases. Clinical signs included behavior changes and erratic movements with loss of equilibrium. Main gross findings were swelling of the gills with white mucus exudate. Microscopically, gills had mild to moderate multifocal necrotizing and lympho-plasmocytic inflammation with hypertrophy and fusion of secondary lamellae. In addition, numerous intralesional trophonts of P. pillulare and cysts of Henneguya spp. were noted. According to our knowledge, this seems to be the first description of Piscinoodinium pillulare e Henneguya spp. infection in fishes reared in Southwestern Goiás, Brazil.

Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water / Produção de anticorpos policlonais anti-Cryptosporidium e padronização da imunofluorescência direta para a detecção de oocistos na água

INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20 percent of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

INTRODUÇÃO: A produção de anticorpos policlonais anti-Cryptosporidium e sua utilização na imunofluorescência para determinar a presença de Cryptosporidium em água são descritas no presente trabalho. MÉTODOS: Dois coelhos foram imunizados com antígeno solúvel e particulado provenientes de oocistos purificados de Cryptosporidium. O soro produzido foi preparado para a extração de imunoglobulinas G, que foram purificadas e conjugadas com isotiocianato de fluoresceína (FITC). Lâminas contendo quantidades conhecidas de oocistos foram preparadas para determinar a sensibilidade da técnica. Para testar a especificidade foram preparadas lâminas contendo cistos de Giardia duodenalis. RESULTADOS: O conjugado foi usado com sucesso em amostras de água contaminadas experimentalmente com oocistos de Cryptosporidium, sendo capaz de detectar até cinco oocistos/spots que corresponde a uma contaminação de 250 oocistos/mL. CONCLUSÕES: As três imunizações realizadas nos coelhos foram suficientes para produção de anticorpos contra Cryptosporidium; a reação de imunofluorescência direta padronizada permitiu a detecção de cinco oocistos em 20 por cento das amostras; não houve reação cruzada com cistos de Giardia duodenalis.

Presença de mosquitos (Diptera: Culicidae) em piscinões na zona leste de São Paulo / The presence of mosquitoes (Diptera: Culicidae) in flood containment reservoirs in the eastern region of the City of São Paulo, Brazil

INTRODUÇÃO: Alterações no ambiente vêm contribuindo com mudanças climáticas, como o aumento do volume de chuvas, que acarreta as inundações. Medidas estão sendo tomadas no enfrentamento das inundações, como a implantação dos reservatórios de contenção de cheias (piscinões). Neste trabalho, foi avaliada a fauna de culicídeos, de importância epidemiológica, nos piscinões Caguaçu e Inhumas. MÉTODOS: Foram realizadas coletas mensais nos piscinões Caguaçu e Inhumas, situados na região leste de São Paulo, de março de 2006 a fevereiro de 2007, empregando-se os métodos de concha entomológica e aspirador. Para análise dos dados, foram realizadas análises estatísticas descritivas e a regressão linear simples. RESULTADOS: Foram coletados 8.917 culicídeos, destacando-se Culex (Culex) quinquefasciatus, que representou 98,9% dos espécimes no Inhumas e 95,2% no Caguaçu. No Caguaçu, a maior frequência de imaturos foi observada no vertedouro (61%) e no Inhumas na canaleta (42,6%). A precipitação prediz 87% da abundância numérica de larvas de terceiro e de quarto estágio no Caguaçu e 60% do número de pupas coletadas. No Inhumas, a precipitação explicou 36% da abundância numérica de larvas e 18% do número de pupas. CONCLUSÕES: Culex quinquefasciatus, vetor de agentes da filariose, arboviroses e fator de incômodo à população, foi a espécie mais frequente nos dois ambientes. Medidas de controle da espécie nos piscinões estudados se fazem necessárias tendo em vista seu potencial epidemiológico.

INTRODUCTION: Alterations in the environment contribute to changes in weather patterns, which cause an increase in rainfall, which causes flooding. Flooding has been addressed by building reservoirs, called piscinões, to contain the excess rain. In this report, we evaluated the Culicidae fauna of epidemiological importance in the reservoirs of Caguaçu and Inhumas. METHODS: Monthly collections were performed in both reservoirs, which are situated in the Eastern section of the City of São Paulo. Monthly collections of Culicidae were undertaken using the entomological scoop method and a battery aspirator during the period between March 2006 and February 2007. Descriptive statistical and simple linear regression analyses of the data were carried out. RESULTS: A total of 8,917 Culicidae mosquitoes were collected. The majority of the insects captured were of Culex (Culex) quinquefasciatus, representing 98.9% of the specimens identified in the Inhumas and 95.2% of those identified in Caguaçu. In the Caguaçu reservoir, the greatest frequency of immature mosquitoes was observed in the spillway (61%). In the Inhumas reservoir, the greatest occurrence of immature Culicidae was detected in the channel (42.6%). The linear regression analysis showed that 87% of the larvae in the third and fourth stages and 60% of the pupae were collected during periods of rainfall. In the Inhumas reservoir, 36% of the larvae and 18% of the pupae were collected during periods of rainfall. CONCLUSIONS: Culex quinquefasciatus, a vector for agents of filariasis and arbovirus diseases and a nuisance to the human population, was frequent in both environments. Methods of control of the species in the studied dams are necessary to reduce their epidemiological potential.

Habitat characteristics for different freshwater snail species as determined biologically through macroin- vertebrate information

Macro-invertebrates including freshwater snails collected from 643 sites over 8 successive seasons among the River Nile, branches, main canals and certain drains in eight Egyptian Governorates. Thirteen snail species and one bivalve species were identified. The most distributed were Lanistus carinatus and Physa acuta while the most abundant were Cleopatra bulimoides and Physa acuta during the whole study. The sites that harbored each snail species in all the examined watercourses were grouped seasonally and their biological assessment was determined by their minimum and maximum total point similarity percentage to that of the corresponded reference site and mean of the total points. Habitats for most snail species attained minimum total point's similarity percentage less than 21% [very poor habitat] during autumn and winter then spring while during summer very poor habitat was harbored by only few snail species. P. acuta was the only survived snails in habitat which attained 0 as a minimum total point's similarity percentage during two seasons and L. carinatus and Succinea cleopatra during one season. With respect to medically important snails very poor sites constituted 23% of Biomphalaria alexandrina sites, 14% of Lymnaea natalensis and 9.4% of Bulinus truncatus sites. The studied macroinvertebrate matrices, total number of organisms, taxa richness, the Ephemeroptera, Plecoptera, and Trichoptera [EPT] index, ratio of EPT index to chironomidae, ratio of scraper to filtering collector, contribution of dominant macroinvertebrate major group, comparison revealed descending tolerances from B. alexanrina followed by L. natalensis then B. truncates, but Hilsenhoff Biotic Index [HBI] showed the same tolerance to organic pollution

A Ten-year Survey of Giardia Cysts in Drinking Water Supplies of Seoul, the Republic of Korea

To understand the distribution of Giardia cysts in drinking water supplies in Seoul, Korea, we collected water samples quarterly at 6 intakes in the Han River, its largest stream and 6 conventional water treatment plants (WTPs) serving drinking water, from 2000 to 2009. Giardia cysts in each of 10 L water were confirmed in 35.0% of intake water samples and the arithmetic mean was 1.65 cysts/10 L (range 0-35 cysts/10 L). The lowest cyst density was observed at Paldang and Kangbuk intakes, and the pollution level was higher at 4 intakes downstream. It seemed that these 4 intakes were under influence of Wangsuk stream at the end of which cysts were found in all samples with the mean of 140 cysts/10 L. The annual mean number of cysts was 0.21-4.21 cysts/10 L, and the cyst level at the second half of the 10 years was about 1/5 of that at first half on average. The cysts were more frequently found in winter, and their mean density was 3.74 cysts/10 L in winter and 0.80-1.08 cysts/10 L in other seasons. All finished water samples collected at 6 WTPs were negative for Giardia in each of 100 L sample for 10 years and cyst removal by physical process was average 2.9-log. It was concluded that conventional water treatment at 6 WTPs of Seoul appears to remove the cysts effectively under the present level of their source water. Domestic wastewater from the urban region could be an important source of Giardia pollution in the river.

Protozoarios en aguas superficiales y muestras fecales de individuos de poblaciones rurales del municipio Montes, estado Sucre, Venezuela / Protozoans in superficial waters and faecal samples of individuals of rural populations of the Montes municipality, Sucre state, Venezuela

En el estado Sucre, el Río Manzanares se ve amenazado por actividades domésticas, agrícolas e industriales originadas por el hombre, convirtiéndose en factor de riesgo ambiental para sus habitantes. En este sentido se planteó evaluar la presencia de protozoarios en aguas superficiales de afluentes del Río Manzanares (Río Orinoco, Quebrada Seca, Río San Juan), municipio Montes, estado Sucre, Venezuela, así como también el análisis de muestras fecales de los habitantes de poblados aledaños. Se recolectaron muestras de aguas superficiales de los afluentes en estudio y fecales entre mayo 2006-abril 2007. Las muestras de aguas superficiales se procesaron con sedimentación por centrifugación, floculación y tinciones de Kinyoun y tricrómica; las muestras fecales se sometieron a examen directo con solución salina fisiológica y lugol, Ritchie modificado y las coloraciones antes mencionadas. Los protozoarios observados con mayor frecuencia en las aguas superficiales en los afluentes fueron: Amebas, Blastocystis sp., Endolimax sp., Chilomastix sp. y Giardia sp. Mientras que Blastocystis hominis, Endolimax nana y Entaomeba coli fueron los de mayor frecuencia observada en las muestras fecales. Los habitantes de Orinoco La Peña resultaron ser los más afectados por las infecciones parasitarias (77,60%), seguido de Río San Juan con 46,63%, y Quebrada Seca con 39,49%. La presencia de protozoarios patógenos y no patógenos en las aguas superficiales demuestra la contaminación fecal de los afluentes evaluados, por lo que representa un foco de infección permanente para los individuos que viven en las cercanías de estas aguas, esto se refleja por la observación de los mismos parásitos en ambas muestras.

In Sucre state, the Manzanares river is threatened by domestic, agricultural and industrial activities, becoming an environmental risk factor for its inhabitants. In this sense, the presence of protozoans in superficial waters of tributaries of the Manzanares river (Orinoco river, Quebrada Seca, San Juan river), Montes municipality, Sucre state, as well as the analysis of faecal samples from inhabitants of towns bordering these tributaries were evaluated. We collected faecal and water samples from may 2006 through april 2007. The superficial water samples were processed after centrifugation by the direct examination and floculation, using lugol, modified Kinyoun and trichromic colorations. Fecal samples where analyzed by direct examination with physiological saline solution and the modified Ritchie concentration method and using the other colorations techniques above mentioned. The most frequently observed protozoans in superficial waters in the three tributaries were: Amoebas, Blastocystis sp, Endolimax sp., Chilomastix sp. and Giardia sp. Whereas in faecal samples, Blastocystis hominis, Endolimax nana and Entaomeba coli had the greatest frequencies in the three communities. The inhabitants of Orinoco La Peña turned out to be most susceptible to these parasitic infections (77.60%), followed by San Juan River (46.63%) and Quebrada Seca (39.49%). The presence of pathogenic and nonpathogenic protozoans in superficial waters demonstrates the faecal contamination of the tributaries, representing a constant focus of infection for their inhabitants, inferred by the observation of the same species in both types of samples.

A Survey of Cryptosporidium Oocysts in Water Supplies during a 10-Year Period (2000-2009) in Seoul

This study has been conducted to estimate the occurrence of Cryptosporidium oocysts in water supplies in the Metropolitan area of Seoul, South Korea, for 10 years from 2000 to 2009. Water samples were collected quarterly at 6 intakes in the Han River and its largest stream and 6 conventional Water Treatment Plants (WTPs) serving drinking water for 10 million people of Seoul. Cryptosporidium oocysts were found in 22.5% of intake water samples and arithmetic mean was 0.65 oocysts/10 L (range 0-22 oocysts/10 L). Although the annual mean of oocyst number was as low as 0.04-1.90 oocysts/10 L, 3 peaks in 2004 and 2007 were observed and the pollution level was a little higher in winter. The lowest density was observed at Paldang intake and the pollution level increased at Kuui and Jayang intakes. At the end of the largest stream, oocysts were found in 70% of collected samples (mean 5.71 oocysts/10 L) and it seemed that its joining the Han River resulted in the increase at Kuui intake and downstream. Oocyst removal by physical process exceeded 2.0-2.3 log and then all finished water samples collected at 6 WTPs were negative for Cryptosporidium in each 100 L sample for 10 years. These results suggested that domestic wastewater from the urban region could be a source of Cryptosporidium pollution and separating sewage systems adjacent to the intakes could be meaningful for some intakes having weakness related to parasitological water quality.

study on biological control of six fresh water snails of medical and veterinary importance

This study evaluated the molluscicidal effect of Commiphora molmol oil extract [Myrrh], on control of six fresh water snails [Lymnaea natalensis, Bilinus truncatus, Biomphalaria alexandrina, Physa acuta, Melania tuberculata and Cleopatra bulimoides]. Also, the extract effect on the egg masses of L. natalensis, B. truncatus, B. alexandrina and Ph. acuta was evaluated. Snails and egg masses were exposed at 16-20°C to various concentrations [conc.]- LD[50] after 24 hours exposure were 264/132, 283/195, 230/252, 200/224, 241/246 and 241/246 ppm for young/adult of L natalensis, B. truncatus, B. alexandrina, Ph. acuta, M. tuberculata and C. bulimoides respectively. LD[100] after 24 hours exposure were 400/400 for L. natalensis, B. truncatus, B. alexandrina, M. tuberculata and C. bulimoides, and 300/300 for Ph. acuta. Also, complete mortality [100%] was achieved for the egg masses of L. natalensis, B. truncatus, B. alexandrina and Ph. acuta at concentrations of 300, 200, 300 and 400 ppm respectively. Lower concentrations gave the same results after longer exposure. LD[100] of C. molmol oil extract [Myrrh] had a rapid lethal effect on the six snail species and their egg masses in high conc, of 300 and 400 ppm. Commiphora molmol is a promising plant to be included with the candidate plant molluscicides. The oil extract of this plant showed a remarkable molluscicidal activity against used snail species

Endosymbionts of Acanthamoeba Isolated from Domestic Tap Water in Korea

In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the alpha-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.