Explore the vast range of titles available.

Elucidating the specific processes and drivers of community assembly in the host microbiome is essential to fully understand host biology. Toward this goal, an important first step is to describe co-occurrence patterns among different microbial taxa, which can be driven by numerous factors, such as host identity. While host identity can be an important influential factor on co-occurrence patterns, a limited number of studies have explored the relative importance of host identity after controlling for other environmental factors. Here, we examined microbial co-occurrence patterns in four phylogenetically distinct trematode species living within the same snail species, collected concomitantly from the same habitat. Our previous study determined that all these trematodes shared some bacterial taxa, and the relative abundance of microbial taxa differed among trematodes, possibly due to differences in their eco-physiological traits. Here, we specifically predict that pairwise microbial co-occurrence patterns also vary among trematode host species. Our results showed that co-occurrence patterns among eight microbial families varied greatly among the four trematode hosts, with some microbial families co-occurring in some trematode species, whereas no such patterns were observed in other trematodes. Our study suggests that the habitat identity (trematode species) and its associated biotic characteristics, such as physiological and ecological traits, can determine co-occurrence patterns among microbial taxa, with substantial effects on local community composition.

Animal microbiomes have been recognized to have significant impact on animal behaviour, development, and health. For parasites, one of the sources of their associated microbes is likely their hosts, making host identity a potential factor explaining variation of microbial communities among conspecific parasites. Using the host-parasite system including the amphipod Paracalliope fluviatilis , and isopod Austridotea annectens , and their common trematode parasite Maritrema poulini , we hypothesized that host identity has an impact on the diversity and community composition of the microbiomes of M. poulini parasites. We focused on the bacterial component of the trematodes’ microbiomes. We investigated the effect of host identity under both intraspecific (among individual isopods) and interspecific (between amphipods and isopods) host levels and predicted that the effect of host identity can be detected under both host hierarchical levels. We found an effect of host identity on both the diversity and community composition of parasite microbiomes between conspecific isopod hosts, while host identity only had an impact on microbiome community composition when comparing parasites infecting amphipod versus isopod hosts. Our study results show that host identity impacts both parasite microbiome composition and taxonomic diversity, providing indirect evidence that the assembly of parasite microbiomes is not merely random, and that horizontal transmission from the host to the parasite might play a key role in shaping parasite microbiomes.

Digeneans are endoparasitic flatworms with complex life cycles including one or two intermediate hosts (first of which is always a mollusk) and a vertebrate definitive host. Digeneans may harbor intracellular endosymbiotic bacteria belonging to the genus Neorickettsia (order Rickettsiales, family Anaplasmataceae). Some Neorickettsia are able to invade cells of the digenean's vertebrate host and are known to cause diseases of wildlife and humans. In this study we report the results of screening 771 digenean samples for Neorickettsia collected from various vertebrates in terrestrial, freshwater, brackish, and marine habitats in the United States, China and Australia. Neorickettsia were detected using a newly designed real-time PCR protocol targeting a 152 bp fragment of the heat shock protein coding gene, GroEL, and verified with nested PCR and sequencing of a 1371 bp long region of 16S rRNA. Eight isolates of Neorickettsia have been obtained. Sequence comparison and phylogenetic analysis demonstrated that 7 of these isolates, provisionally named Neorickettsia sp. 1-7 (obtained from allocreadiid Crepidostomum affine, haploporids Saccocoelioides beauforti and Saccocoelioides lizae, faustulid Bacciger sprenti, deropegid Deropegus aspina, a lecithodendriid, and a pleurogenid) represent new genotypes and one (obtained from Metagonimoides oregonensis) was identical to a published sequence of Neorickettsia known as SF agent. All digenean species reported in this study represent new host records. Three of the 6 digenean families (Haploporidae, Pleurogenidae, and Faustulidae) are also reported for the first time as hosts of Neorickettsia. We have detected Neorickettsia in digeneans from China and Australia for the first time based on PCR and sequencing evidence. Our findings suggest that further surveys from broader geographic regions and wider selection of digenean taxa are likely to reveal new Neorickettsia lineages as well as new digenean host associations.

Parasitic agents, such as helminths, are the most important biotic factors affecting aquaculture, and the fluke Centrocestus formosanus is considered to be highly pathogenic in various fish species. There have been efforts to control this parasite with chemical helminthicides, but these efforts have had unsuccessful results. We evaluated the anthelmintic effect of 37 strains of Bacillus thuringiensis against C. formosanus metacercariae in vitro using two concentrations of total protein, and only six strains produced high mortality. The virulence (CL50) on matacercariae of three strains was obtained: the GP308, GP526, and ME1 strains exhibited a LC50 of 146.2 μg/mL, 289.2 μg/mL, and 1721.9 μg/mL, respectively. Additionally, these six B. thuringiensis strains were evaluated against the cercariae of C. formosanus; the LC50 obtained from the GP526 strain with solubilized protein was 83.8 μg/mL, and it could be considered as an alternative control of the metacercariae and cercariae of this parasite in the productivity systems of ornamental fishes.

BACKGROUND: Neorickettsia are obligate intracellular bacterial endosymbionts of digenean parasites present in all lifestages of digeneans. Quantitative information on the transmission of neorickettsial endosymbionts throughout the complex life cycles of digeneans is lacking. This study quantified the transmission of Neorickettsia during the asexual reproductive phase of a digenean parasite, Plagiorchis elegans, developing within naturally parasitized lymnaeid pond snails. METHODS: Lymnaea stagnalis snails were collected from 3 ponds in Nelson County, North Dakota and screened for the presence of digenean cercariae. Cercariae were identified to species by PCR and sequencing of the 28S rRNA gene. Neorickettsia infections were initially detected using nested PCR and sequencing of a partial 16S rRNA gene of pooled cercariae shed from each parasitized snail. Fifty to 100 single cercariae or sporocysts were isolated from each of six parasitized snails and tested for the presence of Neorickettsia using nested PCR to estimate the efficiency at which Neorickettsia were transmitted to cercariae during asexual development of the digenean. RESULTS: A total of 616 L. stagnalis were collected and 240 (39%) shed digenean cercariae. Of these, 18 (8%) were Neorickettsia-positive. Six Neorickettsia infections were selected to determine the transmission efficiency of Neorickettsia from mother to daughter sporocyst and from daughter sporocyst to cercaria. The prevalence of neorickettsiae in cercariae varied from 11 to 91%. The prevalence of neorickettsiae in sporocysts from one snail was 100%. CONCLUSION: Prevalence of Neorickettsia infection in cercariae of Plagiorchis elegans was variable and never reached 100%. Reasons for this are speculative, however, the low prevalence of Neorickettsia observed in some of our samples (11 to 52%) differs from the high prevalence of other, related bacterial endosymbionts, e.g. Wolbachia in Wolbachia-dependent filariid nematodes, where the prevalence among progeny is universally 100%. This suggests that, unlike the Wolbachia-filaria relationship, the Neorickettsia-digenean relationship is not obligatory mutualism. Our study represents the first quantitative estimate of the Neorickettsia transmission through the asexual phase of the digenean life cycle.

Ruminants infected by Paramphistomidae flukes shed eggs in the feces, which pass through different stages in the environment until the infective stages (metacercariae) are reached. The activity of the soil fungus Mucor circinelloides on the development of eggs of the rumen fluke Calicophoron daubneyi was presently tested with 3 probes, i.e., in petri plates, feces, and an aqueous environment (tubes). The effect of the fungus was assessed by recording the numbers of undeveloped, nonviable, and embryonated eggs. Nonviable eggs were considered when vacuolization occurred, the inner structures were not clearly observed, the eggshell was broken, or the embryo inside was destroyed. By considering the ability of hyphae of M. circinelloides to develop in the presence of C. daubneyi eggs, attach to their surface, and penetrate and destroy the inner embryo, this ovicidal effect was classified as type 3. After a period of 50 days, the percentage of undeveloped eggs in the feces of infected cattle was 40%; furthermore, 27% of the eggs were nonviable, and 33% were embryonated (1 miracidium inside). The addition of 4 doses of M. circinelloides spores directly onto the feces resulted in 9–31% undeveloped eggs, 38–60% nonviable eggs, and 9–21% embryonated eggs, and no statistical significances were obtained among the different doses. Placing the eggs of C. daubneyi into an aqueous solution containing 107 spores of M. circinelloides/ml for 29 days resulted in 43% undeveloped eggs, 40% nonviable eggs, and 17% embryonated eggs, whereas in the controls, the percentages were 48%, 12%, and 40%, respectively. These data demonstrate the usefulness of the spores of the fungus M. circinelloides in limiting the development of the eggs of the trematode C. daubneyi.

Background Neorickettsia are a group of intracellular α proteobacteria transmitted by digeneans (Platyhelminthes, Trematoda). These endobacteria can also infect vertebrate hosts of the helminths and cause serious diseases in animals and humans. Neorickettsia have been isolated from infected animals and maintained in cell cultures, and their morphology in mammalian cells has been described. However, limited information is available on the morphology and localization of Neorickettsia in the trematode host. Methods We used a Neorickettsia- infected strain of the model trematode Plagiorchis elegans to infect Syrian Golden hamsters to produce adult worms. Ultrastructure of Neorickettsia was assessed by transmission electron microscopy of high pressure freezing/freeze substitution fixed specimens. A Neorickettsia surface protein from P. elegans ( PeN sp-3) was cloned and antibodies against the recombinant protein were used to localize Neorickettsia by immunohistochemistry. Results Ultrastructural analysis revealed moderate numbers of pleomorphic endobacteria with a median size of 600 × 400 nm and characteristic double membranes in various tissue types. Endobacteria showed tubular membrane invaginations and secretion of polymorphic vesicles. Endobacteria were unevenly localized as single cells, or less frequently as small morula-like clusters in the ovary, Mehlis’ gland, vitelline follicles, uterus, intrauterine eggs, testis, cirrus-sac, tegument, intestine and the oral and ventral sucker. Examination of hamster small intestine infected with P. elegans showed many endobacteria at the host-parasite interface such as the oral and ventral sucker, the tegument and the excretory pore. Conclusions We conclude that adult P. elegans trematodes carry Neorickettsia endobacteria in varying numbers in many tissue types that support vertical transmission, trematode to trematode transmission via seminal fluid, and possibly horizontal transmission from trematodes to vertebrate hosts. These means appear to be novel mechanisms of pathogen transmission by endoparasitic worms.

Nosema podocotyloidis n. sp. (Microsporidia, Nosematidae) is described from Podocotyloides magnatestis (Trematoda: Opecoelidae), a parasite of the fish Parapristipoma octolineatum (Teleostei) in the Atlantic Ocean. Electron microscopy reveals that all the stages of the cycle (merogony and sporogony) are diplokaryotic and in direct contact with the cytoplasm of host cells. There is no sporophorous vesicle (pansporoblast). The earliest stages observed are meronts, which have a simple plasmic membrane. Their cytoplasm is granular, rich in ribosomes and contains some sacculi of endoplasmic reticulum. They divide by binary fission into diplokaryotic sporonts. The sporonts have a thick electron-dense wall. Their diplokaryon is slightly less electron-dense than the cytoplasm. The cytoplasm of more advanced sporonts has numerous electron-lucent vesicles. Sporonts with two diplokarya divide by binary fission into diplokaryotic sporoblasts. The older sporoblasts are irregular or elongate and the polar filament is in formation. Their cytoplasm is denser, with ribosomes and lamellae of granular endoplasmic reticulum. The sporoblasts evolve into spores. The mature spores are broadly oval and measure 3.6 (3.1–4.0) × 2.58 (1.8–3.3) μm. Their wall is 100–300 nm thick. The polar tube is isofilar with 11–16 coils, 130–155 nm in diameter and arranged in many layers in the centre of the spore. The polaroplast is divided into two regions: an outer electron-dense cup with granular content and lacking lamellae and an internal region, less electron-dense, composed of irregularly arranged sacs. The posterior vacuole, with an amorphous electron-dense content, is present. The new species is compared with other species of Nosema from trematodes. Nosema podocotyloidis n. sp. (Microsporidia, Nosematidae) est décrit de Podocotyloides magnatestis (Trematoda : Opecoelidae), parasite du poisson Parapristipoma octolineatum (Teleostei) pêché dans l’Océan Atlantique. La microscopie électronique montre que tous les stades de développement (mérogonie et sporogonie) sont diplocaryotiques et en contact direct avec le cytoplasme des cellules hôtes. Il n’y a pas de pansporoblaste. Les plus jeunes stades observés sont des mérontes possédant une membrane plasmique simple. Leur cytoplasme est granulaire, riche en ribosomes et contient quelques saccules de réticulum endoplasmique. Ils donnent, par division binaire, des sporontes diplocaryotiques. Les sporontes sont recouverts d’une paroi épaisse dense aux électrons. Leur diplocaryon est légèrement moins dense que le cytoplasme qui, chez les sporontes âgés, présente de nombreuses vésicules claires aux électrons. Les sporontes avec deux diplocaryons donnent, par division binaire, des sporoblastes avec un diplocaryon. Les sporoblastes âgés sont allongés ou de forme irrégulière et présentent le tube polaire en formation. Leur cytoplasme est plus dense aux électrons et riche en ribosomes libres et en réticulum endoplasmique granulaire. Les sporoblastes évoluent en spores. Les spores matures sont grossièrement ovales et mesurent 3.6 (3.1-4.0) × 2.58 (1.8-3.3) μm. Leur paroi a une épaisseur de 100-300 nm. Le tube polaire est isofilaire, mesure 130-155 nm de diamètre et décrit 11-16 tours de spire disposés en plusieurs couches concentriques au niveau du centre de la spore. Le polaroplaste est divisé en deux régions : une externe, opaque aux électrons avec un contenu granulaire sans lamelles, et une région interne, moins opaque aux électrons, composée de saccules arrangés de façon irrégulière. La vacuole postérieure, avec un contenu amorphe opaque aux électrons est présente. La nouvelle espèce est comparée aux autres espèces de Nosema parasites de Trématodes.

Neorickettsia is a genus of intracellular bacteria endosymbiotic in digeneans that may also invade cells of vertebrates and are known to cause diseases of wildlife and humans. Herein, we report results of screening for Neorickettsia of an extensive collection of DNA extracts from adult and larval digeneans obtained from various vertebrates and mollusks in the United States. Seven isolates of Neorickettsia were detected by PCR and sequenced targeting a 527 bp long region of 16S rRNA. Sequence comparison and phylogenetic analysis demonstrated that four isolates matched published sequences of Neorickettsia risticii . Three other isolates, provisionally named “catfish agents 1 and 2” (obtained from Megalogonia ictaluri and Phyllodistomum lacustri , both parasitic in catfishes) and Neorickettsia sp. (obtained from cercariae of Diplostomum sp.), differed from previously known genotypes of Neorickettsia and are likely candidates for new species. All 7 isolates of Neorickettsia were obtained from digenean species and genera that were not previously reported as hosts of these bacteria. Members of four digenean families (Dicrocoeliidae, Heronimidae, Macroderoididae and Gorgoderidae) are reported as hosts of Neorickettsia for the first time. Our study reveals several new pathways of Neorickettsia circulation in nature. We have found for the first time a Neorickettsia from a digenean (dicrocoeliid Conspicuum icteridorum ) with an entirely terrestrial life cycle. We found N. risticii in digeneans ( Alloglossidium corti and Heronimus mollis ) with entirely aquatic life cycles. Previously, this Neorickettsia species was known only from digeneans with aquatic/terrestrial life cycles. Our results suggest that our current knowledge of the diversity, host associations and circulation of neorickettsiae is far from satisfactory.

The freshwater snail family Thiaridae was studied at five different locations: water sources for the Khek River, Thailand. Snail samples were collected by hand using counts per unit of time sampling method between December 2004 and October 2005. The physico-chemical quality of the water changed with the seasons and affected the sampling areas during both the dry season and the flood season. A total of 9,568 snail samples comprised of 14 species were found. These were 284 Tarebia granifera, 24 Melanoides tuberculata, 86 Thiara scabra, 3,295 Paracrostoma pseudosulcospira pseudosulcospira, 736 P. paludiformis paludiformis, 3,266 P. paludiformis dubiosa, 117 P. morrisoni, 304 Brotia (Brotia) binodosa binodosa, 1,250 B. (Brotia) microsculpta, 146 B. (Senckenbergia) wykoffi, 1 B. (Brotia) pagodula, 5 B. (Brotia) binodosa spiralis, 5 B. (Brotia) insolita and 49 B. (Brotia) manningi. The cercariae were investigated using shedding and crushing methods where they were categorized into two types and five species. The first type, Parapleurolophocercous cercariae, were comprised of Haplorchis pumilio Looss, 1899 and Centrocestus formosanus Nishigori, 1924. The second type, Xiphidiocercariae were comprised of Acanthatrium hitaense Koga, 1953, Loxogenoides bicolor Kaw, 1945 and Haematoloechus similis Looss, 1899. The cercarial infection rates in the above 5 species were 0.1% (5:9,568), 0.2% (15:9,568), 0.3% (24:9,568), 0.4% (37:9,568) and 0.1% (5:9,568), respectively. Five species of snails were susceptible to trematode infections. They were T. granifera, M. tuberculata, T. scabra, P. paludiformis paludiformis and B. (Senckenbergia) wykoffi; infections were found in 26.1% (74:284), 33.3% (8:24), 1.2% (1:86), 0.3% (2:736) and 0.7% (1:146), respectively.