Two new species of Diaphorocleidus and new data on D. affinis (Monogenoidea: Dactylogyridae) parasitizing Bryconops cf. affinis (Characiformes: Iguanodectidae) from the Munim River basin, State of Maranho, northeastern Brazil.

During a study conducted on the helminth fauna of fishes in the middle Munim River basin, State of Maranho, northeastern Brazil, three species (two new) of Diaphorocleidus Jogunoori, Kritsky & Venkatanarasaiah, 2004 are reported from the gills of the piaba Bryconops cf. affinis (Iguanodectidae): Diaphorocleidus affinis (Mizelle, Kritsky & Crane, 1968), D. forcipiformis n. sp. and D. munimensis n. sp. New morphological data, drawings, and geographic distribution were provided for D. affinis. Diaphorocleidus forcipiformis n. sp. is characterized by a male copulatory organ with 2 rings counterclockwise and base forming a small tube with two circular flanges and accessory piece bifurcated, claw-shaped, with parts similar in shape and size, one of them distally bifid. Diaphorocleidus munimensis n. sp. is differentiated by the accessory piece, which is composed of two large pieces, united in their proximal part, claw-shaped, and distally hooked. The findings of this initial survey on the helminth parasites of fishes in this area emphasize the importance of carrying out comparable surveys on other fish species. Such surveys may lead to the identification of unknown species or new occurrences.

A new species of Urocleidoides Mizelle & Price 1964 (Monogenea: Dactylogyridae) parasite of Hemiodus orthonops Eigenmann & Kennedy, 1903 (Hemiodontidae) from the upper Paraná River floodplain.

During the study of ectoparasites (Platyhelminthes) of fish in the floodplain of the Upper Paraná River - Brazil, a new species of Urocleidoides, present in the gill filaments of Hemiodus orthonops (Hemiodontidae), is described using morphological description and molecular data from the mitochondrial region of cytochrome c oxidase, subunit 1 (COI) and the partial region of the 28S rDNA gene. Urocleidoides luquei n. sp. differs from all its congeners by the presence of a filament that joins the accessory piece to the base of the male copulatory organ and resembles U. paradoxus and U. surianoae. Phylogenetic analyses using molecular data revealed that U. luquei n. sp. forms a paraphyletic group concerning the other Urocleidoides species. In this way, as well as contributing to the description of a new species, we seek to encourage and contribute to the increase in research using integrative taxonomy, thus making it possible to elucidate some unresolved questions about the genus Urocleidoides.

Component communities and annual and seasonal variations of metazoan parasites in Eleotris pisonis (Gmelin, 1789) (Gobiiformes: Eleotridae) in the Amazon River, Brazil.

Our hypothesis for this study was that annual and seasonal variations do not influence the structure of the component communities and the diversity of metazoan parasites of spinycheek sleeper (Eleotris pisonis) in the Amazon River, state of Amapá, Brazil. A total of 164 fish were collected between 2020 and 2021, from which 888 parasites were found. In 2020, five species of parasites were found (one Nematoda, one Digenea, one Acanthocephala, one Arachnida and one Crustacea); and in 2021, five species were also found (three Nematoda, one Digenea and one Crustacea). Larvae of Contracaecum sp. were the dominant taxon throughout the study. The parasite species richness and Brillouin diversity index were higher in 2021, without significant differences between seasonal periods. Some component communities of parasites showed differences between years and between seasonal periods. These facts do not support the hypothesis that such variables would not influence the component communities of the parasites. Lastly, this report provides the first records of Spirocamallanus inopinatus, Genarchella genarchella, Acari, Ergasilus sp., Neoechinorhynchus sp., larvae of Pseudoproleptus sp. and larvae of Contracaecum sp. in E. pisonis.

Influence of urbanization on schistosomiasis infection risk in Anhui Province based on sixteen year's longitudinal surveillance data: a spatio-temporal modelling study.

BACKGROUND: Urbanization greatly affects the natural and social environment of human existence and may have a multifactoral impact on parasitic diseases. Schistosomiasis, a common parasitic disease transmitted by the snail Oncomelania hupensis, is mainly found in areas with population aggregations along rivers and lakes where snails live. Previous studies have suggested that factors related to urbanization may influence the infection risk of schistosomiasis, but this association remains unclear. This study aimed to analyse the effect of urbanization on schistosomiasis infection risk from a spatial and temporal perspective in the endemic areas along the Yangtze River Basin in China. METHODS: County-level schistosomiasis surveillance data and natural environmental factor data covering the whole Anhui Province were collected. The urbanization level was characterized based on night-time light data from the Defense Meteorological Satellite Program Operational Linescan System (DMSP-OLS) and the National Polar-Orbiting Partnership's Visible Infrared Imaging Radiometer Suite (NPP-VIIRS). The geographically and temporally weighted regression model (GTWR) was used to quantify the influence of urbanization on schistosomiasis infection risk with the other potential risk factors controlled. The regression coefficient of urbanization was tested for significance (α = 0.05), and the influence of urbanization on schistosomiasis infection risk was analysed over time and across space based on significant regression coefficients. Variables studied included climate, soil, vegetation, hydrology and topography. RESULTS: The mean regression coefficient for urbanization (0.167) is second only to the leached soil area (0.300), which shows that the urbanization is the most important influence factors for schistosomiasis infection risk besides leached soil area. The other important variables are distance to the nearest water source (0.165), mean minimum temperature (0.130), broadleaf forest area (0.105), amount of precipitation (0.073), surface temperature (0.066), soil bulk density (0.037) and grassland area (0.031). The influence of urbanization on schistosomiasis infection risk showed a decreasing trend year by year. During the study period, the significant coefficient of urbanization level increased from - 0.205 to - 0.131. CONCLUSIONS: The influence of urbanization on schistosomiasis infection has spatio-temporal heterogeneous. The urbanization does reduce the risk of schistosomiasis infection to some extend, but the strength of this influence decreases with increasing urbanization. Additionally, the effect of urbanization on schistosomiasis infection risk was greater than previous reported natural environmental factors. This study provides scientific basis for understanding the influence of urbanization on schistosomiasis, and also provides the feasible research methods for other similar studies to answer the issue about the impact of urbanization on disease risk.

Metazoan parasites of Hoplias aff. malabaricus, Trachelyopterus galeatus and Schizodon borellii (Osteichthyes) from the Protected Area and its main tributary, Brazil.

A study of the parasite-host interactions was conducted in the protected area popularly known as the "Refúgio Biológico de Santa Helena" and its tributary, the São Francisco Falso River. For each of the three host species, Schizodon borellii, Hoplias aff. malabaricus, and Trachelyopterus galeatus, 30 fish were collected from 2018 to 2019. A total of 2172 parasites were identified among the three host species. Among these, the Monogenea class had the highest number of species, with 26 taxa, followed by Copepoda with eight taxa, Digenea with six taxa, and Cestoda and Nematoda with one taxon each. Eleven new records of infection/infestation were found among the host species such as Urocleidoides paradoxus, Urocleidoides ramentacuminatus, Rhinoxenus arietinus, Mymarothecioides sp. (Monogenea), Ergasilus cf. bryconis, Lernaea devastatrix, and Brasergasilus sp. (Copepoda) parasitizing S. borellii. Trinigyrus sp., Vancleaveus sp. (Monogenea), Gamispinus diabolicus (Copepoda) present in T. galeatus, and Gamidactylus jaraquensis (Copepoda) present in H. aff. malabaricus. This study contributes to the record of parasite species occurrence in the vicinity of a protected area and helps fill gaps in the knowledge of fish parasitic fauna in the Neotropical region.

Molecular evidence of the absence of Metagonimus yokogawai (Katsurada, 1912) in Europe: report of Metagonimus sp. in cyprinoid fish from the River Danube in Hungary.

Trematodes of the genus Metagonimus Katsurada, 1912 (Digenea: Heterophyidae) are zoonotic parasites that cause infections in humans, with most cases reported in Southeast Asia. Larvae from the second intermediate host, called metacercariae, of one of human-infecting species, M. yokogawai (Katsurada, 1912), have been reported from cyprinoid fish in Europe. In the present study, we provided DNA-based evidence that metacercariae of Metagonimus, which are commonly found in the scales of various cyprinoids in Central Europe (Danube River in Hungary) do not belong to M. yokogawai. Sequence analysis of the ITS region, 28S rDNA, and cox1 genes showed that this species is clearly distinct from all Asian species, including M. yokogawai, which probably does not occur in Europe. Metacercariae from cyprinoids might belong to Metagonimus romanicus (Ciurea, 1915), an insufficiently known species described from Romania.

Endoparasite Communities of Fish at Different Trophic Levels in the Western Brazilian Amazon: Human, Environmental and Seasonal Influence.

PURPOSE: The composition of the fish parasite community depends on several factors related to the environment, the host and its biology. This study aimed to evaluate the influence of environmental factors in anthropized and conserved areas on the endoparasite community structure in fish at different trophic levels, in addition to verifying that some species of Digenea are indicators of conserved environments. METHODS: The study was carried out in the Upper Juruá River region, Western Amazon, Brazil. Six sampling sites were selected in this region and grouped in conserved and degraded environments. Fish were caught from periods of drought and flood, using passive and active sampling methods. Fish collected were measured, weighed, necropsied and the parasites found were counted, fixed, and subjected to morphological analysis. Physical and chemical variables and environmental characteristics were measured in all sites. RESULTS: The present study demonstrated that environmental variables in a floodplain system can influence the richness, diversity, composition and abundance of endoparasites in hosts at different trophic levels. In addition, anthropized environments may favor the abundance of some generalist parasites and present a more homogeneous biota between seasonal periods compared to conserved environments. CONCLUSION: Study contributed with information supporting the importance of conservation of aquatic environments, and demonstrated that fish parasites can be excellent indicators of environments.

Fish parasites as proxy bioindicators of degraded water quality of River Saraswati, India.

The nature and intensity of water pollution determine the effects on aquatic biota and aquatic ecosystem health. The present study aimed at assessing the impact of the degraded physicochemical regime of river Saraswati, a polluted river having a historical legacy, on the parasitic infection and the role of fish parasite as a bioindicator of water quality. Two Water Quality Indices (WQIs) were adopted as useful tools for assessing the overall water quality status of polluted river based on 10 physicochemical parameters. Total 394 fish (Channa punctata) were examined. Ectoparasite Trichodina sp., Gyrodactylus sp., and endoparasites Eustrongylides sp. were collected from the host fish. Prevalence, mean intensity and abundance for each sampling period were calculated for the determination of parasitic load. The parasitic load of Trichodina sp. and Gyrodactylus sp. was significantly (p < 0.001) higher in winter, whereas the parasitic load of Eustrongylides sp. showed no significant (p > 0.05) seasonal fluctuation. The parasitic load of ectoparasites was negatively correlated with temperature, free carbon dioxide, biochemical oxygen demand, and WAWQI but positively correlated with electrical conductivity and CCMEWQI. Fish health was found to be adversely affected by degrading water qualities and parasitic infection. A 'vicious cycle' develops as a result of the interplay among deteriorating water quality, withering fish immunological defence, and amplifying parasitic infection. Since parasitic load was strongly conditioned by the combined influence of a suite of water quality parameters the fish parasites can be used as a powerful indicator of deteriorating water quality.

Seasonal patterns of infestation by monogenean parasites of fish and their relationship with water parameters in two rivers with different disturbance gradients in southeastern Brazil.

Here, we evaluate the relationships between the infestation rates of five monogenean parasites species with the dry and wet seasons, with the organic and inorganic parameters of the water of two rivers: the Jacaré-Pepira and Jacaré-Guaçú, and with the condition factors of its fish hosts: Serrasalmus maculatus and Astronotus crassipinnis, in the state of São Paulo, southeastern Brazil. Fish were collected between January and December 2017. Anacanthorus serrasalmi, Amphithecium speirocamarotum and Gussevia asota had higher abundance rates (Student's t test, p ≤ 0.05) in the wet season. Gussevia asota had its abundance negatively correlated to nitrate in the Jacaré-Pepira River and with total nitrogen and potassium in the Jacaré-Guaçú River. Regarding the fish hosts condition factors, was observed a positive correlation with the abundances of G. asota in the Jacaré-Guaçú River, and with A. serrasalmi in the Jacaré-Pepira River. In general, wet season favored an increasing in the infestation rates of the monogeneans parasites in their host species, mainly in the river considered as the most polluted, the Jacaré-Guaçú River. Of the five parasites species analyzed in this study, only Gussevia astronoti and Rhinoxenus piranhus had no interaction with seasonality, river water variables, or fish host condition factors. On the other hand, G. asota had interactions both with water parameters (nitrate and total nitrogen) and with the hosts condition factors, which reflected in the abundance and intensity rates, showing itself as a species sensitive to changes in the environment and, therefore, that can be considered as a bioindicator organism.

Myxobolus makombensis n. sp. infection in African carp Labeobarbus batesii from the Makombè River, Cameroon: morphological and molecular characterization.

In this study, we examined myxozoan infections of Labeobarbus batesii sampled from the Makombè River in Cameroon. Fish were infected with Myxobolus makombensis n. sp. in the gill filament and M. dibombensis in the fins. Mature myxospores of M. makombensis n. sp. are pyriform in frontal view and biconvex in lateral view, with a truncated and slightly narrow anterior end. Spore dimensions (mean ± SD, with range in parentheses) are 17.5 ± 0.22 (16.2-18.9) µm length, 13.4 ± 0.25 (12-14.9) µm width, and 7 ± 0.21 (6.7-7.5) µm thickness, and spores exhibit a conspicuous anterior intercapsular appendix of 4.4 ± 0.18 (3.9-5.5) µm length. Myxospores have 2 pyriform polar capsules of unequal size; the larger one is 9.8 ± 0. 22 (8.2-10.9) µm long × 4.7 ± 0.15 (3.5-5.2) µm wide, and the smaller one is 8.8 ± 0.22 (7-10) µm long × 4.3 ± 0.12 (3.5-5.2) µm wide. Polar filaments possess 10 to 11 coils in the large polar capsule and 8 to 10 coils in the small polar capsule. Phylogenetic analysis of SSU rDNA sequences showed clustering of M. makombensis n. sp. close to M. dibombensis recently reported from the fins of the same host within a clade composed exclusively of parasites infecting cyprinid fishes.

Morphological and Molecular Characterization of Myxobolus nkondjockei sp. nov. (Myxozoa: Myxobolidae), A Parasite of Labeo batesii Boulenger, 1911 (Teleostei: Cyprinidae) from Makombè River in Cameroon.

PURPOSE: Myxozoans are economically important group of metazoan parasites, which can cause diseases in a large variety of commercially important fishes. Increased knowledge on molecular features has shown that traditional descriptive characters may be misleading. Combination of both descriptive and molecular features is therefore necessary for an integrated taxonomic assessment. METHODS: Cyprinid Labeo batesii, sampled in the Makombè River at Nkondjock in Cameroon were examined for myxosporeans. Identification of parasite species was based on morphological and molecular sequence analyses of myxospores. Phylogenetic analysis was performed using maximum likelihood (ML) and Bayesian inference (BI) methods. RESULTS: The scales of L. batesii were infected by Myxobolus nkondjockei sp. nov Their mature myxospores are ovoid in frontal view and lenticular in lateral view, with two rounded ends. These myxospores measured 10.3 (10-10.9) µm length and 8.0 (7.3-8.5) µm width. Myxospores have two ovoid and equal sizes polar capsules. They measured 4.5 (4.0-5.0) µm in length and 2.4 (2-2.9) µm in width. Polar tubules were coiled in 4-5 turns perpendicular to the longitudinal axis of the polar capsules. Phylogenetic analysis of the 18S rDNA sequence show clustering of M. nkondjockei sp. n. close to an undetermined species Myxobolus sp. reported infecting gill lamellas of Labeo rohita from India. CONCLUSION: The morphological, molecular and phylogenetic data provided for M. nkondjockei sp. n. are solid basis for further identification of this myxozoan of which pathogenicity probably plays an economic role at culturing the hosts.

Temporal and seasonal variation of metazoan parasites in Pimelodus ornatus (Siluriformes: Pimelodidae) from the Amazon River, Brazil.

This study investigated the influence of temporal and seasonal (i.e., rainy and dry seasons) in communities and infracommunities of metazoan parasites in Pimelodus ornatus, a Siluriformes from the Amazon River in the state of Amapá, in northern Brazil. A total of 144 fish were collected during 2020 and 2021. In 2020, five species of parasites were found (1 Monogenea, 3 Nematoda, and 1 Cestoda), and in 2021, seven species of parasites were found (1 Monogenea, 3 Nematoda, 1 Cestoda, 1 Digenea, and 1 Crustacea). In 2020, Spirocamallanus inopinatus was dominant, while in 2021, Contracaecum sp. was dominant. Brillouin diversity, species richness, and evenness were higher in 2021. There were differences for some parasite infracommunities between years and between seasonal periods. Brillouin diversity and species richness were higher in the rainy season, but there were no differences in evenness and Berger-Parker dominance between the seasonal periods. Temporal and seasonal variations were mainly influenced by rainfall levels, oxygen levels, and environmental temperature and, later, by the availability of infective stages of parasites in the environment and the size of the hosts. Lastly, the influence of the temporal and seasonal variations on the structure of communities and infracommunities of parasites was weaker than expected due to the low infection rates of the majority of parasites found.

Cosmetocleithrum infinitum sp. n. (Monogenoidea: Dactylogyridae) parasite of the gills of Anadoras grypus (Siluriformes: Doradidae) from the Amazonas river, in Loreto, Peru.

Among Siluriformes, fishes of the family Doradidae harbor dactylogyrids from Cosmetocleithrum Kritsky, Thatcher and Boeger, 1986. Anadoras grypus (Cope) (Siluriformes, Doradidae) locally named in Peru as "rego-rego" hosts a new species of Cosmetocleithrum described herein as Cosmetocleithrum infinitum sp. n. The male copulatory organ (MCO) and vagina of the new species reveal its morphological uniqueness among members of Cosmetocleithrum. The MCO in the new species is an elongate, sclerotized sinuous tube, with 12 expanded coils, with counterclockwise orientation; the accessory piece is sclerotized, comprising a kind of cone, with a large opening at its base, with a branch in the posterior middle part and with pointed ends. The vagina is a sclerotized long, winding, coiled tube with a cone-shaped opening.

RECRUITMENT AND SEASONAL OCCURRENCE OF PARASITES IN JUVENILE INVASIVE ROUND GOBIES (NEOGOBIUS MELANOSTOMUS) IN THE ST. LAWRENCE RIVER, QUEBEC, CANADA.

The round goby (Neogobius melanostomus) is an invasive species that has become one of the most abundant fish in the St. Lawrence River, Quebec, Canada over the past 15 yr. Since its introduction, the round goby has acquired a number of native parasites, yet little is known about the dynamics of parasite recruitment. To examine this question, young-of-the-year and juvenile round gobies were collected monthly from 2 localities in the river (Îles de la Paix, Île Dorval) from June through November 2012. At Îles de la Paix, round gobies (n = 180) were infected with 3 species of parasites, all larval stages (Diplostomum spp., Tylodelphys scheuringi, Neoechinorhynchus tenellus). Prevalence of the digenean Diplostomum spp. varied from 3.3 to 13.3%, and mean abundance from 0.03 to 0.53 from June through September, with a maximum in August. The digenean T. scheuringi was seen only in August, at a prevalence of 10.0% and a mean abundance of 0.53. The acanthocephalan N. tenellus was observed in June, August, and September, prevalence ranging from 3.3 to 10.0% and mean abundance from 0.03 to 0.27. Maximum infection for all 3 species occurred in August. All infected fish were ≥44 mm in total length (TL). Fish infected with more than 1 parasite species were >60 mm TL. No round goby (n = 178) was infected at Île Dorval. This study demonstrated that the invasive round goby starts to acquire parasite infections in the St. Lawrence River in the first year of life and may contribute to the transmission of some parasites within this ecosystem.

Endoparasite fauna of freshwater fish from the upper Juruá River in the Western Amazon, Brazil.

The Amazon region may present a high diversity of endoparasites with a high degree of endemism. In this sense, this study describes the endoparasite fauna in freshwater fish from the Upper Juruá, in the Western Amazon. The study was carried out around the municipalities of Cruzeiro do Sul, state of Acre, and Guajará, state of Amazonas, Brazil. Fish were caught between periods of droughts and floods, using passive and active sampling methods. In the laboratory, specimens were biometrically analysed and necropsied. As a result, a total of 23,740 endoparasites were recorded, belonging to 62 species, with 91 new host reports and 91 new occurrences for the Western Amazon. Nematoda and Digenea were the most diverse and abundant groups, and the increase in host fish richness and diversity influenced the diversity and richness of endoparasites in the environments. In this sense, the present study expands the number of new reports, and contributes data on the distribution and richness of endoparasites for South America.

Composition and structure of the parasitic fauna of Hypostomus spp. (Loricariidae: Hypostominae) from a Neotropical river in Brazil.

Hypostomus are abundant in Brazilian rivers and streams. In the Ivaí River, the loricariids represent 20.3% of the total species of the basin. Of these 13 species belong to Hypostomus. However, to date, there are no studies on these fish parasitic fauna. Thus, this research aimed to analyze the distribution of the parasitic infracommunity of six species of Hypostomus from the Ivaí River and investigate how the infracommunity is structured in these hosts. One hundred and twenty-eight fish were analyzed, belonging to six sympatric species of Hypostomus (Hypostomus hermanni, H. cochliodon, H. albopunctatus, H. regani, Hypostomus sp.1, and Hypostomus sp.2); of these, 92.9% were parasitized with at least one taxon, totaling 1478 specimens of parasites. The parasitic fauna was composed of the ectoparasites Trinigyrus anthus, T. carvalhoi, Unilatus unilatus (monogeneans), and Placobdella spp. (hirudinea), and the endoparasites Austrodiplostomum compactum (digenean) and Procamallanus annipetterae (nematode). The parasites exhibited similar patterns of infection in all hosts, including a low number of species, low diversity, and numerical dominance of a group of parasites. However, permutational multivariate analysis of variance (PERMANOVA) showed different parasite species compositions among the hosts. Hypostomus cochliodon and H. regani had the highest parasite richness, while Hypostomus sp.1 and Hypostomus sp.2 showed low abundance and intensity of parasitic infections. However, Hypostomus sp.1 showed the highest values of evenness, although the parasite composition in both species did not differ. The results presented herein contribute to increasing the knowledge about the parasitic fauna of Hypostomus spp. from the Ivaí River by presenting new hosts and locality records.

Cryptosporidiosis outbreak in Amazonia, French Guiana, 2018.

BACKGROUND: Cryptosporidiosis outbreaks in South America are poorly documented. In March 2018, 51 cases of cryptosporidiosis were reported in Maripasoula, a village located in a remote forest area along the border between Surinam and French Guiana. METHOD: To identify the origin of the epidemic, we performed epidemiological, microbiological, and environmental investigations. Only the cases involving diarrhoea and Cryptosporidium-positive stool were considered as bona fide, while cases involving diarrhoea and close contact with a confirmed case were classified as "possible". RESULTS: We identified 16 confirmed cases and 35 possible ones. Confirmed cases comprised nine children (median age of 18 months, range: 6-21), one immunocompromised adult and six soldiers. One child required a hospitalisation for rehydration. All 16 Cryptosporidium stools were PCR positive, and sequencing of the gp60 gene confirmed only one Cryptosporidium hominis subtype IbA10G2. Tap water consumption was the only common risk factor identified. Contamination of the water network with Cryptosporidium parvum subtype IIdA19G2 was found. CONCLUSION: Water quality is a major public health issue in Amazonian French Guiana, especially for population at risk (children, people with comorbidity, travelers). For them, alternative water supply or treatment should be implemented.

Spatial Variability in Streambed Microbial Community Structure across Two Watersheds.

Both spatial and temporal variability are key attributes of sedimentary microbial communities, and while spatial effects on beta-diversity appear to dominate at larger distances, the character of spatial variability at finer scales remains poorly understood, especially for headwater stream communities. We investigated patterns of microbial community structure (MCS) in biofilms attached to streambed sediments from two watersheds across spatial scales spanning <1 m within a single stream to several hundred kilometers between watersheds. Analyses of phospholipid fatty acid (PLFA) profiles indicated that the variations in MCS were driven by increases in the relative abundance of microeukaryotic photoautotrophs and their contribution to total microbial biomass. Furthermore, streams within watersheds had similar MCS, underscoring watershed-level controls of microbial communities. Moreover, bacterial community structure assayed as either PCR-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprints or PLFA profiles edited to remove microeukaryotes indicated a distinct watershed-level biogeography. No distinct stream order-level distributions were identified, although DGGE analyses clearly indicated that there was greater variability in community structure among 1st-order streams than among 2nd- and 3rd-order streams. Longitudinal gradients in microbial biomass and structure showed that the greatest variations were associated with 1st-order streams within a watershed, and 68% of the variation in total microbial biomass was explained by sediment atomic carbon-to-nitrogen ratio (C:N ratio), percent carbon, sediment surface area, and percent water content. This study confirms a distinct microbial biogeography for headwater stream communities driven by environmental heterogeneity across distant watersheds and suggests that eukaryotic photoautotrophs play a key role in structuring bacterial communities on streambed sediments. IMPORTANCE Microorganisms in streams drive many biogeochemical reactions of global significance, including nutrient cycling and energy flow; yet, the mechanisms responsible for the distribution and composition of streambed microbial communities are not well known. We sampled sediments from multiple streams in two watersheds (Neversink River [New York] and White Clay Creek [WCC; Pennsylvania] watersheds) and measured microbial biomass and total microbial and bacterial community structures using phospholipid and molecular methods. Microbial and bacterial community structures displayed a distinct watershed-level biogeography. The smallest headwater streams within a watershed showed the greatest variation in microbial biomass, and 68% of that variation was explained by the atomic carbon-to-nitrogen ratio (C:N ratio), percent carbon, sediment surface area, and percent water content. Our study revealed a nonrandom distribution of microbial communities in streambeds, and showed that microeukaryotic photoautotrophs, environmental heterogeneity, and geographical distance influence microbial composition and spatial distribution.

Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample.

Acanthamoeba spp. are opportunistic human pathogens that cause granulomatous amoebic encephalitis and keratitis, and their accurate detection and enumeration in environmental samples is a challenge. In addition, information regarding the genotyping of Acanthamoeba spp. using various PCR methods is equally critical. Therefore, considering the diverse niches of habitats, it is necessary to develop an even more efficient genotyping method for Acanthamoeba spp. detection. This study improved the sensitivity of detection to avoid underestimation of Acanthamoeba spp. occurrence in aquatic environmental samples, and to accurately define the pathogenic risk by developing an efficient PCR method. In this study, a new nested genotyping method was established and compared with various PCR-based methods using in silico, lab, and empirical tests. The in silico test showed that many PCR-based methods could not successfully align specific genotypes of Acanthamoeba, except for the newly designed nested PCR and real-time PCR method. Furthermore, 52 water samples from rivers, reservoirs, and a river basin in Taiwan were analysed by six different PCR methods and compared for genotyping and detection efficiency of Acanthamoeba. The newly developed nested-PCR-based method of genotyping was found to be significantly sensitive as it could effectively detect the occurrence of Acanthamoeba spp., which was underestimated by the JDP-PCR method. Additionally, the present results are consistent with previous studies indicating that the high prevalence of Acanthamoeba in the aquatic environment of Taiwan is attributed to the commonly found T4 genotype. Ultimately, we report the development of a small volume procedure, which is a combination of recent genotyping PCR and conventional real-time PCR for enumeration of aquatic Acanthamoeba and acquirement of biologically meaningful genotyping information. We anticipate that the newly developed detection method will contribute to the precise estimation, evaluation, and reduction of the contamination risk of pathogenic Acanthamoeba spp., which is regularly found in the water resources utilised for domestic purposes.

Species diversity and distribution of schistosome intermediate snail hosts in The Gambia.

There is a need for recent information on intermediate snail hosts of schistosomes in The Gambia; the previous studies were conducted over three decades ago. This study assessed the incidence, species diversity, distribution and infection status of schistosome intermediate snail hosts in the country. Malacological surveys were conducted in all 5 regions of The Gambia: Central River Region (CRR), Upper River Region (URR), Western Region (WR), Lower River Region (LRR) and North Bank Region (NBR). Sampling of snails was undertaken at 114 sites that included permanent water bodies such as streams (bolongs), rice fields, irrigation canals and swamps; and temporal (seasonal) laterite pools. Ecological and physicochemical factors of sites were recorded. Snails were identified morphologically and screened for schistosome infections using molecular techniques. Freshwater snails were found at more than 50% (60/114) of sites sampled. While three species of Bulinus were collected, no Biomphalaria snails were found in any of the sites sampled. Of the total 2877 Bulinus snails collected, 75.9% were identified as Bulinus senegalensis, 20.9% as Bulinus forskalii and 3.2% as Bulinus truncatus. Seasonal pools produced the largest number of snails, and CRR was the region with the largest number of snails. Bulinus senegalensis was found more in seasonal pools as opposed to permanent sites, where B. forskalii and B. truncatus were observed to thrive. Bulinus snails were more common in seasonal sites where aquatic vegetation was present. In permanent sites, the abundance of snails increased with increase in water temperature and decrease in water pH. Bulinus senegalensis was found infected with both S. haematobium and S. bovis, while B. forskalii and B. truncatus had only S. bovis infection. While the human parasite S. haematobium was restricted to just four sites, the livestock parasite S. bovis had a much more widespread geographical distribution across both CRR and URR. This new information on the distribution of intermediate snail hosts of schistosomes in The Gambia will be vital for the national schistosomiasis control initiative.