Bacterial flora of the schistosome vector snail Biomphalaria glabrata

The aerobic heterotrophic bacterial flora in over 200 individuals from 10 wild populations and 3 laboratory colonies of the schistosome vector snail Biomphalaria glabrata was examined. Internal bacterial densities were inversely proportional to snail size and were higher in stressed and laboratory-reared snails. The numerically predominant bacterial genera in individual snails included Pseudomonas, Acinetobacter, Aeromonas, Vibrio, and several members of the Enterobacteriaceae. Enterobacteriaceae seldom predominated in laboratory colonies. Our data suggest that Vibrio extorquens and a Pasteurella sp. tend to predominate in high-bacterial-density snails. These snails may be compromised and may harbor opportunistic snail pathogens.     

Tissue invasion of laboratory-reared Biomphalaria glabrata by a harpacticoid copepod

Copepods were observed in the tissues of 3 of 23 Biomphalaria glabrata snails examined histologically. All were heavily encapsulated by hemocytes and were dead. The copepods are most likely members of the order Harpacticoida, based on external morphology. This tissue invasion appears to be accidental rather than symbiotic or predatory, but could be a cause of observed mortality in laboratory snail colonies.     

Effects of snail size and age on the prevalence and intensity of avian schistosome infection: relating laboratory to field studies

Both the prevalence and intensity of patent infection by avian schistosomes (Trichobilharzia ocellata) increase with increasing size of lymnaeid snails (Stagnicola elrodi) collected in Flathead Lake, Montana. Because the size and age of a snail are positively correlated, snails of different sizes may have experienced differential duration of exposure to and development of infection. Another possibility is that infection itself induces snail gigantism. Each of these possibilities could lead to increased prevalence and intensity of infection among the oldest-largest snails. To decouple size variation from many correlated effects of age and to test for parasite-induced gigantism, laboratory experiments standardized snail size-age-at-infection, exposure history, inoculating dose, and duration of infection. The positive relationship between size and prevalence was eliminated in the laboratory, but the relationship between size and infection intensity remained. Laboratory results thus suggest that infection intensity is related to snail size per se, whereas prevalence in the field is related to snail size only through the correlation between size and age. In addition, under these experimental conditions, infected snails were no larger than uninfected snails, so the patterns observed in the field might not be attributable to gigantism.     

The hemolymph of Biomphalaria snail vectors of schistosomiasis supports a diverse microbiome

The microbiome - the microorganism community that is found on or within an organism's body - is increasingly recognized to shape many aspects of its host biology and is a key determinant of health and disease. Microbiomes modulate the capacity of insect disease vectors (mosquitoes, tsetse flies, sandflies) to transmit parasites and disease. We investigate the diversity and abundance of microorganisms within the hemolymph (i.e. blood) of Biomphalaria snails, the intermediate host for Schistosoma mansoni, using Illumina MiSeq sequencing of the bacterial 16S V4 rDNA. We sampled hemolymph from five snails from six different laboratory populations of B. glabrata and one population of B. alexandrina. We observed 279.84 ± 0.79 amplicon sequence variants per snail. There were significant differences in microbiome composition at the level of individual snails, snail populations and species. Snail microbiomes were dominated by Proteobacteria and Bacteroidetes while water microbiomes from snail tank were dominated by Actinobacteria. We investigated the absolute bacterial load using qPCR: hemolymph samples contained 2784 ± 339 bacteria/μl. We speculate that the microbiome may represent a critical, but unexplored intermediary in the snail–schistosome interaction as hemolymph is in very close contact with the parasite at each step of its development.     

An efficient system for markerless gene replacement applicable in a wide variety of enterobacterial species

We describe an improved allelic-exchange method for generating unmarked mutations and chromosomal DNA alterations in enterobacterial species. Initially developed for use in Salmonella enterica, we have refined the method in terms of time, simplicity, and efficiency. We have extended its use into related bacterial species that are more recalcitrant to genetic manipulations, including enterohemorrhagic and enteropathogenic Escherichia coli and Vibrio parahaemolyticus. Data from over 50 experiments are presented including gene inactivations, site-directed mutagenesis, and promoter exchanges. In each case, desired mutations were identified by polymerase chain reaction screening typically from as few as 10-20 colonies up to a maximum of 300 colonies. The method does not require antibiotic nor nutritional markers in target genes and works efficiently in wild-type strains, obviating the need for specialized hosts or genetic systems. The use is simple, requiring basic laboratory materials, and represents an alternative to existing methods for gene manipulation in the Enterobacteriaceae.     

Supercooling ability in two populations of the land snail Helix pomatia (Gastropoda: Helicidae) and ice-nucleating activity of gut bacteria

The land snail Helix pomatia (Gastropoda: Helicidae) is widely distributed in Northern and Central Europe where it may experience subzero temperatures during winter months. Its supercooling ability was studied in two populations of H. pomatia. One population originated from Southern Sweden (Gotaland) and the other from Central France (Auvergne). In the experimental design, they were acclimated, over 2 weeks, to artificial winter conditions (hibernation, T=5 degrees C). The Swedish snails showed a rather limited supercooling ability (temperature of crystallization, T(c)=-6.4+/-0.8 degrees C), significantly greater, however, than the supercooling capacity of the population from France (T(c)=-4.6+/-1.4 degrees C). In artificial spring conditions (3 months of hibernation followed by a progressive acclimation, over 2 weeks, to activity at T=20 degrees C), both populations exhibited a similar high T(c) (-2.0+/-1.0 degrees C). The lower T(c) of hibernating Swedish snails could be due to a greater loss of body water, accompanied by a higher concentration of solutes in the hemolymph. In both populations, the variation in hemolymph osmolality measured between hibernating (250-270 mOsm kg(-1)) and active (165-215 mOsm kg(-1)) snails may be explained by the variation in body water mass and did not suggest the production of colligative cryoprotectants. Moreover, the three bacterial strains, Buttiauxella sp., Kluyvera sp., and Tatumella sp. (Enterobacteriaceae) which were isolated from fed snails, but absent in starved snails, did not show any ice-nucleating activity at temperatures higher than -9 degrees C. Only the strain Kluyvera sp. initiated nucleation at -9 degrees C. This strain, therefore, is a weak, also termed a Type III or Class C ice-nucleating active bacterium, but with no influence on the supercooling ability of individual snails. In summary, fluctuations in body water mass of hibernating snail populations, triggering changes in osmolyte concentration, rather than the presence of endogenous ice-nucleating-active bacteria, accounts for fluctuations in their T(c).     

Cercarial shedding of Echinostoma cinetorchis and experimental infection of the cercariae to several kinds of snails

The development of Echinostoma cinetorchis in several snail species reared in laboratory aquaria was observed. The eggs from adult flukes collected from the intestine of rats were cultivated to miracidia, and exposed to Hippeutis sp. snails. Observations were made for cercarial shedding from the exposed snails. The cercariae shed from the snails were again exposed to several species of fresh water snails in order to observe metacercarial formation in the snails and their infectivity to final hosts. The results obtained in this study were as follows: 1. Twenty miracidia were exposed to each snail of Hippeutis sp. About 58.3% of the above snails (7 out of 12) were dead before shedding the cercariae, and the remainder shed the cercariae for a period of 7 to 9 days before death. 2. Cercarial shedding from the infected snails started from the 25th day after the exposure to miracidia, and the total number of cercariae shed per snail was 684 in average (range; 482-904). 3. The size of rediae developed in the infected Hippeutis sp. snails was 1,242 x 214 microns in average, and the number of rediae per snail was 350 in average (range; 120-510). 4. About 40 to 50 cercariae shed from the Hippeutis sp. snails were each exposed to several species of snails reared in the laboratory. The metacercarial formation was confirmed by dissecting the infected snails, 12 to 16 days after the infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gut bacterial communities in the giant land snail Achatina fulica and their modification by sugarcane-based diet

The invasive land snail Achatina fulica is one of the most damaging agricultural pests worldwide representing a potentially serious threat to natural ecosystems and human health. This species is known to carry parasites and harbors a dense and metabolically active microbial community; however, little is known about its diversity and composition. Here, we assessed for the first time the complexity of bacterial communities occurring in the digestive tracts of field-collected snails (FC) by using culture-independent molecular analysis. Crop and intestinal bacteria in FC were then compared to those from groups of snails that were reared in the laboratory (RL) on a sugarcane-based diet. Most of the sequences recovered were novel and related to those reported for herbivorous gut. Changes in the relative abundance of Bacteroidetes and Firmicutes were observed when the snails were fed a high-sugar diet, suggesting that the snail gut microbiota can influence the energy balance equation. Furthermore, this study represents a first step in gaining a better understanding of land snail gut microbiota and shows that this is a complex holobiont system containing diverse, abundant and active microbial communities.     

Leech predation on juvenile freshwater snails: effects of size,species and substrate

Summary The leechGlossiphonia complanata does not appear to have substantial impact on snail populations, but this may be due to most studies focusing on adult snails rather than juvenile snails. In this study I investigated how predation rates ofG. complanata feeding on newly-hatched and juvenile snails was affected by snail species, snail size, snail density and substrate, in a laboratory experiment. Number of snails eaten increased with increasing density resulting in a type II functional response curve. Predation rates were higher when leeches were feeding onLymnaea emarginata than onPhysa gyrina, whereas there was no significant difference in predation rates when they were feeding onL. emarginata andHelisoma anceps. Sandy substrates and greater snail size resulted in decreased predation rates. Sand reduced movement speed ofG. complanata, which probably reduced encounter rates. Thus, there was a comparatively large effect of leech predation on newly-hatched snails, due to a high probability of encounter and high predation rates, but spatial and temporal refuges probably reduce the importance of leech predation as a structuring force in freshwater snail assemblages.     

Save your host,save yourself? Caste‐ratio adjustment in a parasite with division of labor and snail host survival following shell damage

Shell damage and parasitic infections are frequent in gastropods, influencing key snail host life‐history traits such as survival, growth, and reproduction. However, their interactions and potential effects on hosts and parasites have never been tested. Host–parasite interactions are particularly interesting in the context of the recently discovered division of labor in trematodes infecting marine snails. Some species have colonies consisting of two different castes present at varying ratios; reproductive members and nonreproductive soldiers specialized in defending the colony. We assessed snail host survival, growth, and shell regeneration in interaction with infections by two trematode species, Philophthalmus sp. and Maritrema novaezealandense, following damage to the shell in the New Zealand mud snail Zeacumantus subcarinatus. We concomitantly assessed caste‐ratio adjustment between nonreproductive soldiers and reproductive members in colonies of the trematode Philophthalmus sp. in response to interspecific competition and shell damage to its snail host. Shell damage, but not parasitic infection, significantly increased snail mortality, likely due to secondary infections by pathogens. However, trematode infection and shell damage did not negatively affect shell regeneration or growth in Z. subcarinatus; infected snails actually produced more new shell than their uninfected counterparts. Both interspecific competition and shell damage to the snail host induced caste‐ratio adjustment in Philophthalmus sp. colonies. The proportion of nonreproductive soldiers increased in response to interspecific competition and host shell damage, likely to defend the parasite colony and potentially the snail host against increasing threats. These results indicate that secondary infections by pathogens following shell damage to snails both significantly increased snail mortality and induced caste‐ratio adjustments in parasites. This is the first evidence that parasites with a division of labor may be able to produce nonreproductive soldiers according to environmental factors other than interspecific competition with other parasites.     

Zebra mussels decrease burrowing ability and growth of a native snail, <Emphasis Type="Italic">Campeloma decisum</Emphasis>

Invasive species can drive native organisms to extinction by limiting movement and accessibility to essential resources. The purpose of this study was to determine if zebra mussels (Dreissena polymorpha) affect the burrowing ability and growth rate of a native snail, Campeloma decisum. Snails with and without zebra mussels were collected from Douglas Lake, MI, and burrowing depths were studied in both the laboratory and Douglas Lake. Growth rates were calculated as the amount of shell growth from 2004 to 2005. Both the tendency of snails to burrow and the depth to which they burrowed into the substrate were significantly decreased by the presence of zebra mussels on snail shells in both laboratory and lake experiments. There was no difference in the percentage of snails that exhibited growth as a function of zebra mussel presence. However, for those snails that grew, there was a 50% higher growth rate for snails without zebra mussels compared to snails with zebra mussels. These negative effects of zebra mussels on growth and burrowing ability will likely lead to decreases in snail population densities in the future. Handling editor: S. Wellekens     

Periphytic food and predatory crayfish: relative roles in determining snail distribution

Summary In the laboratory and field, we examined how periphyton (food of snails) and predatory crayfish influenced snail distribution in Trout Lake, a permanent, northern Wisconsin lake. Laboratory experiments (with no crayfish) tested the importance of periphyton biomass in determining snail preference among rocks, and among rock, sand, and macrophyte substrates. Among rocks with four different amounts of periphyton, periphyton biomass and the number of Lymnaea emarginata, Physa spp., and Amnicola spp. were positively related. A similar, but non-significant, trend occurred for Helisoma anceps. A field experiment at a site in Trout Lake where predation risk was low confirmed the preference by snails for periphyton covered rocks; more snails colonized rocks with periphyton than rocks without. When given a choice of rock, sand, and macrophytes in the laboratory, L. emarginata preferred high periphyton biomass and rock. Laboratory and field results contrasted with the distribution of snails in Trout Lake; no snails occurred in areas with abundant periphyton-covered rocks, but snails were abundant nearby on scattered rocks with little periphyton. However, where snails were absent, crayfish were abundant (14.5 crayfish-trap^–1-day^–1), and where snails were abundant, crayfish were rare (3.2 crayfish-trap^–1-day^–1), suggesting that crayfish predation reduced snails. The hypothesis that the negative association between snail and periphyton biomass resulted from snail grazing was supported by the results of a field snail enclosure-exclosure experiment (1 m² cages; n=3). All experiments and observations therefore suggest that: 1) crayfish predation is more important than a preference for high periphyton biomass in determining snail distribution in Trout Lake; 2) periphyton biomass is negtively related to snail grazing; and 3) crayfish had a positive indirect effect on periphyton by preying on grazing snails.     

Trade-off between mating and predation risk in the marine snail, Littorina plena

Abstract. We investigated the hypothesis that predation risk affects mating decisions in the intertidal snail Littorina plena in Bamfield Inlet, Northeast Pacific. First, we conducted a field tethering experiment to test the assumption that mating pairs of snails are more susceptible to predation than solitary individuals, and then performed a laboratory experiment to quantify the effect of predation threat on the propensity of snails to form mating pairs. Our results support the hypothesis, in that "mating pairs" were more frequently killed than single snails in the field, and snails were less likely to form mating pairs in the laboratory when simulated predation risk was high (chemical cues from crushed conspecifics were added to the water) than when it was low (no risk cues were added to the water). In contrast to several earlier studies, we found no effect of individual size on snail susceptibility to predation, perhaps because our two size classes were contiguous and snails within them were not dissimilar enough. The results of the behavioral experiment were consistent with this lack of individual size effect on snail vulnerability; both size classes of snails showed a significant and similar tendency to decrease mating when predation risk was high. Taken together, the results of this and recent studies indicate that predators can considerably affect the behavior of littorinid snails, including their movement patterns, feeding, and reproduction. We argue that greater consideration should be given to how marine invertebrates trade off predation risk and activities related to reproduction.     

Effect of a leaf-shredding invertebrate on organic matter dynamics and phosphorus spiralling in heterotrophic laboratory streams

Summary The effect of invertebrate shredders on organic matter dynamics and phosphorus spiralling was studied over a 30-week period in laboratory streams. The streams were fed by groundwater, layered with cobble and gravel from a natural stream, covered with opaque material to eliminate algal growth, and initially contained 195 g/m² of autumn-shed leaves. Four weeks after leaf addition, leaf-shredding snails (Goniobasis clavaeformis) were added to each of three streams in densities of 75, 220, and 800/m². A fourth stream contained no snails and served as a control.Presence of snails increased the loss rates of coarse particulate organic matter (CPOM) and total organic matter (TOM), primarily by increasing leaf fragmentation and seston export. Although snail feeding increased specific metabolism of microbes associated with CPOM and cobble surfaces, it was not enough to compensate for reduction in bacterial cell numbers per unit surface area and in stream TOM. Consequently mineralization of detritus and whole stream phosphorus utilization rate were maximum in the stream with no snails and decreased with increasing snail density. From previous simulations of a stream model based on the nutrient spiralling concept, we predicted that there should be an intermediate shredder density which would minimize phosphorus spiralling length (maximize phosphorus utilization) in a natural stream nearby. Our current results conflict with the model-based predictions primarily because the increase in microbial metabolism was less important than reduction in bacterial cell numbers and total benthic organic matter resulting from snail feeding. Although our results indicate macroinvertebrate shredders reduce phosphorus utilization in headwater streams, shreders may increase nutrient utilization downstream where riparian inputs are lower, thus linking low- and high-order streams.Research supported by the National Science Foundation's Ecosystem Studies Program under Interagency Agreement No. BSR-8103181, A02 with the U.S. Department of Energy, under Contract No. DE-AC05-840R21400 with Martin Marietta Energy Systems, Inc.Publication No. 2394, Environmental Sciences Division, ORNL     

Ecology of bacterial communities in the schistosomiasis vector snailBiomphalaria glabrata

The internal colony-forming bacterial flora of the schistosome intermediate host snailBiomphalaria glabrata (Say) has been characterized in ca. 500 individual snails from Puerto Rico, Guadeloupe, and St. Lucia, and from laboratory aquaria. Freshly captured wild snails harbor 2–40×10⁶ CFU·g^–1, and healthy aquarium snails harbor 4–16×10⁷ CFU·g^–1, whereas moribund individuals have 4–10 times as many bacteria as healthy individuals from the same habitats.Pseudomonas spp. are the most common predominant bacteria in normal snails, whereasAcinetobacter, Aeromonas, andMoraxella spp. predominate in moribund snails. External bacterial populations in water appear to have little effect on the composition and size of the flora in any snail. In addition to normal (healthy) and moribund snails, a third group of snails has been distinguished on the basis of internal bacterial density and predominating genera. These high-density snails may have undergone stresses and may harbor opportunistic pathogens. The microfloras of wild and laboratory-reared snails can be altered and stimulated to increase in density by crowding the snails or treating them with antibiotics.     

Prevalence of schistosome infections within molluscan populations: observed patterns and theoretical predictions

The paper draws together a large and scattered body of empirical evidence concerning the prevalence of snail infection with schistosome parasites in field situations, the duration of the latent period of infection in snails (and its dependence on temperature), and the mortality rates of infected and uninfected snails in field and laboratory conditions. A review and synthesis of quantitative data on the population biology of schistosome infections within the molluscan host is attempted and observed patterns of infection are compared with predictions of a schistosomiasis model developed by May (1977) which incorporates differential snail mortality (between infected and uninfected snails) and latent periods of infection. It is suggested that the low levels of prevalence within snail populations in endemic areas of schistosomiasis are closely associated with high rates of infected snail mortality and the duration of the latent period of infection within the mollusc. In certain instances, the expected life-span of an infected snail may be less than the duration of the latent period of infection. Such patterns generate very low levels of parasite prevalence. A new age prevalence model for schistosome infections within snail populations is developed and its predictions compared with observed patterns. The implications of this study of observed and predicted patterns of snail infection within molluscan populations are discussed in relation to the overall transmission dynamics of schistosomiasis.     

Intermediate host specificity in Schistosoma mansoni

Miracidia of Schistosoma mansoni penetrate into many kinds of snails, but development of normal sporocysts takes place only in certain species of Biomphalaria. Different populations of this snail vary greatly in laboratory infection rates with S. mansoni originating from diverse geographic localities. Cross-exposure experiments show that compatibility factors exist in both snails and parasites. Susceptibility of stocks of Biomphalaria to particular strains of S. mansoni is genetically determined and may be modified by selection in the laboratory. In a compatible snail, the sporocyst develops without host tissue reaction; in incompatible snails the early larvae are rapidly surrounded by amebocytes and fibroblasts, and destroyed. This reaction resembles the generalized host cellular response elicited by any foreign body. An individual snail exposed to many miracidia may have both developing and encapsulated sporocysts side by side within its tissues. The weight of current evidence suggests that elicitation or absence of this cellular response resides in the recognition or nonrecognition of the sporocyst as a foreign body. The sporocyst tegument surface, which forms within a few hours after miracidial penetration, may have a molecular conformation identical with that of the snail, or may be able to bind specific host molecules, so that detection and subsequent encapsulation by host cells are averted. Presuming genetic determination of the sporocyst surface structure and of the host cell detection capability, differing infection rates would result from the particular frequencies of relevant genes in the populations concerned.     

半褶织纹螺(Nassarius semiplicatus)及其生活环境中分离培养的细菌毒性测试与种类分析

在中国沿海地区,因食用织纹螺导致的中毒事件时有发生.最近的研究表明,河豚毒素及其衍生物是织纹螺中主要的致毒成分.但是,对于织纹螺中河豚毒素的来源还不清楚.[目的]本研究尝试分离、培养和鉴定织纹螺及其生活环境中的细菌,并对其毒性进行分析,为探明织纹螺中河豚毒素的可能来源提供科学依据.[方法]先后于2006年6月13日和19日在江苏省盐城采集织纹螺样品,应用小鼠生物法对织纹螺样品的毒性进行了测试;从织纹螺体内及其生活环境中分离细菌,并选择部分菌株进行了室内培养;以直接竞争酶联免疫分析方法(ELISA)对培养菌株中的河豚毒素进行了检测;通过对细菌16S核糖体DNA(rDNA)部分序列的测定,对有毒菌株进行了初步的种灯分析.[结果]实验结果表明,采集的织纹螺为半褶织纹螺,两次采集样品的毒性分别为247 MU(mouse unit,小鼠单位)和270MU/100g组织(湿重).对14个菌株进行了毒性检测,其中有毒细菌9株.产毒菌株的毒性普遍较低,毒性范围为15～96 ng/g.有毒菌株核糖体序列与弧菌(Vibrio)、希瓦氏菌(Shewanella)、动性球菌(Planococcus)、海单胞菌(Marinomonas)、发光杆菌(Photobacterium)等菌属有较高的相似性,可能具有较近的亲缘关系.[结论]研究发现半褶织纹螺体内及其生活环境中存在能够产生河豚毒素的细菌,说明织纹螺中的河豚毒素可能与其体内及其生活环境中的细菌有关,有必要进行深入研究.     

Ecological factors in schistosome transmission, and an environmentally benign method for controlling snails in a recreational lake with a record of schistosome dermatitis

The avian schistosomes, Trichobilharzia stagnicolae, T. physellae and Gigantobilharzia sp., that cause Schistosome Dermatitis (Swimmers' Itch) in humans were studied in the laboratory and at Cultus Lake, British Columbia, Canada in relation to the biology and behavior of their intermediate snail hosts, Stagnicola catascopium, Physa sp. and Gyraulus parvus, respectively, and their definite bird hosts. Wind-driven, surface currents were measured. Populations of snails, close to host-bird roosting logs had a very high prevalence of schistosome infections. An experiment that mechanically disturbed the epilithic habitat of the snails using a boat-mounted rototiller or a tractor and rake, eliminated almost all of the snails if the disturbance was done in areas of high snail concentration in shallow areas of the lake during the breeding and early development phase of the snail. It is proposed that the incorporation of snail habitat disturbance into management programs is an effective way to control Schistosome Dermatitis.     

Transplantation of Schistosoma japonicum daughter sporocysts in Oncomelania hupensis

Schistosoma japonicum daughter sporocysts obtained from infected Oncomelania hupensis hupensis were successfully transplanted to parasite-free O. hupensis hupensis. Survival and infection rates of recipient snails were 80% and 75% respectively. Intramolluscan development of transplanted daughter sporocysts in recipient snails appears to proceed in a similar manner as those reported for transplanted S. mansoni and S. haematobium in their respective snail intermediate hosts. Complete colonization of the digestive gland of recipient snails by sporocysts was observed 80 days after transplantation. Cercarial production during a 10-day observation from recipient snails was characterized by periods of high and low and irregular daily emissions. The average daily cercarial production was 150 per snail. Cercariae produced by recipient snails were infective to mice. Of those cercariae exposed to mice, approximately 30% developed to adult schistosomes. These results have definitive utility in the maintenance of S. japonicum in the laboratory.