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Chemotherapy-based control of schistosomiasis haematobia. 3. Snail studies monitoring the effect of chemotherapy on transmission in the Msambweni area, Kenya.

Regular snail sampling was performed at 40 sites, representing the principal snail habitats, during a 4 year chemotherapy programme targetted at school-children in the Msambweni area of the coastal plain of Kenya. Populations of Bulinus africanus group snails, primarily from pools, showed seasonal variations, dropping when sites dried out and rising when they were refilled by the rains. Transmission, judged by the recovery of snails shedding typical fucocercous cercariae, continued throughout the treatment period at very low levels (less than 1% of the snails collected were infected) with peaks in October/November and in January/February after seasonal rains. Spatially, most infected snails were recovered from 2 large pools near the sea, close to the school that responded least well to the initial chemotherapy programme. Later, infected snails were found sporadically in inland pools, rice fields and temporary streams near another school where there was evidence of substantial reinfection as the study progressed. The snail findings are consistent with a reduction but not elimination of transmission associated with the chemotherapy programme.

Animals↗

Feeding clusters and olfaction in the mangrove snail Terebralia palustris (Linnaeus) (Potamididae: Gastropoda).

Large numbers of the snail Terebralia palustris (Linnaeus) (Potamididae; Gastropoda) are frequently observed feeding in a cluster on a single fallen mangrove leaf, yet none are present on leaves nearby. Consequently, we investigated the food-finding ability of T. palustris in a Kenyan mangrove forest using field experiments. We estimated the attractive effect of different cues and analysed the paths (video-recorded) of snails when approaching a food-related odour. This intertidal snail can potentially use both air-borne and water-borne odours to locate food. T. palustris is attracted to conspecifics feeding on leaves, while intact leaves as well as non-feeding snails are not attractive. Moreover, the guiding stimulus appears to be compounds released when the leaves are damaged.T. palustris also seems able to discriminate between different food items; it is more strongly attracted to green mangrove leaves than senescent or fallen ones or mangrove propagules, probably because green leaves release a greater amount of attractive cues.Feeding snails thus recruit more snails to feed on the same leaf. The ecological implications of this behaviour are discussed: a large number of snails on the same leaf counteracts the ability of crabs to remove the leaf being eaten by the snails.

Journal Article↗

Effects of infection with Echinostoma paraensei on the circulating haemocyte population of the host snail Biomphalaria glabrata.

Circulating haemocytes from Echinostoma paraensei-infected M line Biomphalaria glabrata snails, or from age- and size-matched control snails, were studied on plastic slides with phase-contrast optics. Granulocytes, hyalinocytes, and round cells were consistently present; granulocytes were further categorized as 'fully spread' (FS) or 'partially spread' (PS). Among control snails, the relative percentage and estimated number/mm3 of round cells declined significantly with increased snail size, and the corresponding values for both categories of granulocytes increased. At 1 day post-infection (p.i.) with E. paraensei, overall composition of the haemocyte population was relatively unaffected, but by 8 days p.i. infected snails had significantly higher relative percentages of round cells and PS granulocytes than controls. Because a marked increase in the number of circulating haemocytes is also evident by 8 days p.i., infected snails had approximately 12 times more round cells and PS granulocytes/mm3 of haemolymph than did controls. At 30 days p.i. the relative and absolute abundance of PS granulocytes was still significantly elevated, but otherwise haemocyte populations did not differ from control snails. Alterations in granulocyte size in infected snails were also noted. Infection with E. paraensei has a striking impact of circulating haemocyte populations and also increases the relative concentration of haemocytes with less ability to adhere to a foreign surface.

Animals↗

Effects of temperature and host density on the snail-finding capacity of cercariae of Echinostoma caproni (Digenea: Echinostomatidae).

The effect of temperature (19-36 degrees C) and snail host density (0.014-10 snails/l) on the snail-finding capacity of Echinostoma caproni cercariae is described. The initial swimming speed increased whereas the length of the infective period decreased with increasing temperature. The combined effect resulted in the E. caproni cercarial snail-finding capacity being temperature independent in the range 19 to 36 degrees C at a snail density of 0.014 snails/l. A moderate temperature dependency was, however, seen at a lower snail density. The cercarial snail-finding capacity was snail-host density dependent in the density range 0.014 to 1 snail/l. The findings from this study show that a relatively low and biologically realistic snail host density must be used in experimental studies if realistic estimates of the dynamics of cercarial transmission are to be obtained.

Analysis of Variance↗

Effects of parasitism and pesticide exposure on characteristics and functions of hemocyte populations in the freshwater snail Lymnaea palustris (Gastropoda, Pulmonata).

Morphological characteristics and functions of hemocytes were used to compare the immunological effects of biological and chemical stress in the freshwater snail Lymnaea palustris. Animals were either infected by a trematode parasite (Metaleptocephalus sp.), or exposed to environmental contaminants, namely atrazine and hexachlorobenzene (HCB). Three populations of circulating hemocytes, morphologically and cytochemically distinct (round cells, hyalinocytes, granulocytes), were identified in both control and parasitized or pesticide-exposed snails. After 6 h of exposure, HCB and atrazine resulted in 8-fold increases in the mean total number of hemocytes, whereas only a 2.2-fold increase was observed 6 h after cercaria emission in parasitized snails. The impact of HCB was limited to the first 24 h of exposure, whereas long-lasting effects of atrazine were observed. Hyalinocytes and, to a lesser extent, round cells contributed most to the increases in hemocyte density in pesticide-exposed snails. Parasitism and atrazine treatment resulted in significant increases of lectin-stained hemocytes, whereas exposure to HCB did not affect the percentages of stained and unstained cells. Hemocyte phagocytic activity increased in HCB-exposed snails but with no concomitant change of the oxidative burst. Opposite results were obtained in atrazine-treated snail hemocytes, with unchanged phagocytosis and decreased phorbol 12-myristate 13-acetate-stimulated production of reactive oxygen intermediates. No increase in phagocytosis, or in the production of reactive oxygen intermediates, was observed in hemocytes from parasitized snails. Infection with the immunologically compatible trematode parasite Metaleptocephalus sp. and exposure to atrazine generated similar reactions from circulating hemocytes, whereas a different response pattern was observed in HCB-exposed snails.

Animals↗

Taste discrimination in conditioned taste aversion of the pond snail Lymnaea stagnalis.

Conditioned taste aversion (CTA) in the pond snail Lymnaea stagnalis has been widely used as a model for gaining an understanding of the molecular and behavioral mechanisms underlying learning and memory. At the behavioral level, however, it is still unclear how taste discrimination and CTA interact. We thus examined how CTA to one taste affected the feeding response induced by another appetitive food stimulus. We first demonstrated that snails have the capacity to recognize sucrose and carrot juice as distinct appetitive stimuli. We then found that snails can become conditioned (i.e. CTA) to avoid one of the stimuli and not the other. These results show that snails can distinguish between appetitive stimuli during CTA, suggesting that taste discrimination is processed upstream of the site where memory consolidation in the snail brain occurs. Moreover, we examined second-order conditioning with two appetitive stimuli and one aversive stimulus. Snails acquired second-order conditioning and were still able to distinguish between the different stimuli. Finally, we repeatedly presented the conditional stimulus alone to the conditioned snails, but this procedure did not extinguish the long-term memory of CTA in the snails. Taken together, our data suggest that CTA causes specific, irreversible and rigid changes from appetitive stimuli to aversive ones in the conditioning procedure.

Animals↗

Effects of copper sulfate toxicity on cercariae and metacercariae of Echinostoma caproni and Echinostoma trivolvis and on the survival of Biomphalaria glabrata snails.

Copper in the form of copper sulfate (CuSO4) decreases the survival of Biomphalaria glabrata snails, but the effects of this molluscicide on Echinostoma caproni and Echinostoma trivolvis, 2 species of digeneans that use B. glabrata as intermediate hosts, are not known. Studies were done on the effects of various concentrations of CuSO4 in artificial spring water (ASW) on the survival and infectivity of E. caproni and E. trivolvis cercariae. Solutions containing 1.0, 0.1, and 0.01% CuSO4 were 100% lethal within 2 hr of exposure for both species. Time to 50% mortality in 0.001% CuSO4 was 8 hr for E. caproni and 16 hr for E. trivolvis; at 24 hr, the controls showed 50 and 65% mortality, respectively. Treatment of cercariae of both species for 0.5 hr in 0.001% CuSO4 had no effect on the ability of cercariae to form normal cysts in juvenile B. glabrata snails. However, treatment with 0.01% CuSO4 for 0.5 hr caused a significant reduction in the ability of cercariae of both species to encyst in snails. Treatment of encysted metacercariae of both species in 0.001% CuSO4 for I hr had no effect on subsequent excystation of these echinostomes in a trypsin-bile salts medium, whereas concentrations of 1.0, 0.1, and 0.01% CuSO4 and 1.0 and 0.1% CuSO4 decreased chemical excystation of E. caproni and E. trivolvis cysts, respectively. Survival studies on the effects of CuSO4 in Locke's solution on chemically excysted metacercariae of both species were also done. Excysted metacercariae of both species were killed by 2 hr in either 0.1 or 0.01% CuSO4 in Locke's solution. However, time to 50% mortality for both species of excysted metacercariae in 0.001% CuSO4 was approximately 5 hr. Time to 50% mortality for the controls was about 12 hr. Survival of juvenile B. glabrata snails was also examined. All B. glabrata snails were dead by 6 hr in 1 and 0.1% CuSO4 in ASW. Biomphalaria glabrata snails showed 50% mortality by about 6 hr in 0.01% CuSO4 and about 80% were still alive at 24 hr in 0.001% CuSO4. All controls were alive at 24 hr, at which time the experiment was terminated. Concentrations greater than 0.001% CuSO4 increased snail mortality, as well as that of the cercariae and excysted metacercariae of E. caproni and E. trivolvis. Our findings suggest that concentrations of copper sufficient to eliminate juvenile B. glabratta snails are also sufficient to kill the cercariae and excysted metacercariae of these digeneans but not the encysted metacercariae, which may be protected by their cyst walls.

Analysis of Variance↗

Schistosoma mansoni infection in diapausing Biomphalaria glabrata snails: studies of temperature and genetic influences on diapausing behavior.

Snails that are capable of undergoing diapause can circumvent unfavorable environmental conditions, including long periods of drought. Studies were performed to investigate possible temperature and/or genetic factors that may trigger lamella formation and diapausing behavior. The influence of diapause in Biomphalaria glabrata snails on susceptibility to Schistosoma mansoni infection and levels of cercarial production was also investigated. Rearing temperatures of 23 degrees C or higher did not induce lamella formation or diapause, regardless of the parental phenotype. However, substantial percentages of progeny from lamellated or lamellated/diapausing parental snails developed lamellae at 18 degrees C and underwent diapause. Only a small percentage of offspring from nonlamellated parents formed lamellae at this temperature. Juvenile snails exposed just prior to diapause, or immediately following a diapause period of three weeks, were highly susceptible to infection by S. mansoni miracidia. Snails that underwent diapause produced comparable or only slightly fewer cercariae than did nondiapausing snails. These studies indicate that diapause in B. glabrata does little to decrease a snail's ability to act as an intermediate host for S. mansoni or to interrupt the development of the parasite. For these reasons, we believe that greater attention should be given to diapausing snail populations when planning field surveys or mollusciciding programs.

Animals↗

A polymerase chain reaction assay for detecting snails infected with bilharzia parasites (Schistosoma mansoni) from very early prepatency.

In the present study, we adapted a polymerase chain reaction (PCR) assay, previously shown by us to be very sensitive for detecting cercariae in water, for the sensitive detection of Schistosoma mansoni DNA in infected snails from early prepatency. Polymerase chain reaction primers were designed based on the 121-basepair highly repeated sequence we previously identified in the genome of S. mansoni. The DNA was prepared from the snails by a simple alkaline extraction procedure, and the PCR assay enabled a clear differentiation between infected and normal snails. Infected snails were detected as early as one day after penetration of a single miracidium. The high sensitivity of the test enabled identification of a single infected snail even when its DNA was pooled with material from up to 99 uninfected snails, thus demonstrating the possibility of mass diagnosis in pools of snails. The assay has the potential for large-scale determination of prepatent infection prevalence in snails, thus offering new possibilities for the evaluation of schistosomiasis transmission and for schistosomiasis control, as discussed.

Animals↗

Observations on the host-parasite relations between Echinostoma revolutum and lymnaeid snails.

Echinostoma revolutum from Taiwan was studied in lymneid snails at 29 +/- 0.5 degrees C. Given 3-5 miracidia, 95% of Lymnaea ollula and 40% of Lymnaea swinhoei became positive; the prepatent periods were 18 and 25 days, respectively. The following are based on the observations in Lymnaea ollula: The time required for miracidial penetration was about one hour. The sporocysts developed only in the ventricle of the snail but mother rediae developed in the heart and other organs. Mature daughter rediae were not found in the heart cavity. The sporocysts reached the ventricle within 3 days. Mother rediae were released after 6 days and daughter rediae after 8 days. Given 5 miracidia, 1-3 sporocysts reached the heart and 2-20 mother rediae were found per snail. The number of mature daughter rediae was usually 100-200 although more than a thousand may develop in a snail. The sporocysts and mother rediae attained maximal size 9 days postinfection and started degeneration 13 days postinfection. Daughter rediae were largest at the beginning of cercarial emergence and decreased in size thereafter. Simultaneous production of daughter rediae and cercariae by the mother redia was seen only once in this snail mature cercariae were obtained in 10 days postinfection. The cercariae emerged from a small area of mantle collar near the posterior corner of shell aperture. They were negatively phototactic and positively geotactic. An estimation showed that each snail shed about 350 cercariae a day. The cercariae reached the pericardial cavity of snail in one hour via the renal orifice and metacercariae were seen 4-5 hours after exposure. The infectivity of cercariae at various times after shedding, as expressed by cyst recovery rates, were: 51.6%, O-hr old; 76.1%, 2-hr; 68% 4-hr; 32%, 6-hr; 3%, 8 and 10-hr. Cyst recovery rates were not different within the dosage of 50-500 cercariae per snail. Most metacercariae recovered 1-2 days after cercarial exposure were viable; only 5 among 6,533 cysts were dead.

Echinostoma↗

Infection of two lymnaeid snails with Fasciola gigantica in Giza, a field study.

Population density of Lymnaea natalensis and L. columella and their natural infection with Fasciola gigantica were followed in two water bodies in Abu Rawash area, Giza Governorate over two years. L. natalensis snails were more abundant during December-February, while L. columella snails had high density during autumn. According to the snail size, we found that the reproduction of the two vector snails occurred in summer and early autumn. Fasciola was present in 0.6 and 2.0% of the examined L. natalensis and L. columella snails respectively. A high prevalence of F. gigantica in L. columella snails occurred in summer and early autumn, while their presence in L. natalensis was irregular. Naturally infected Lymnaea snails, with Fasciola, are usually of large size (> 10mm) of a given population sample. Other three types of trematode cercariae were found in both snail species; Xiphidiocercariae, Echinostomes and Amphistomes.

Animals↗

Studies on the epidemiology of schistosomiasis in Liberia: the prevalence and intensity of schistosomal infections in Bong County and the bionomics of the snail intermediate hosts.

Urine samples from 3548 individuals residing in six of the eight districts which comprise Bong County, Liberia, the project area of the Bong County Agricultural Development Project (BCADP), and fecal specimens from 3408 of these individuals were examined for schistosome ova. A total of 164 water sites, including rice paddies, were surveyed for schistosome vector snails and monthly changes in snail population density and infection rate were determined in selected water sites. Bulinus globosus was more widely distributed than Biomphalaria pfeifferi but the latter species showed a higher infection prevalence (12.3%) than the former one (10.3%). Snail population density and infection rate fluctuated with season, being higher in the dry season and lower during periods of heavy rainfall. Dessication and/or heat stress may have contributed to the contraction of snail population size at the end of the dry season. More water sites contained infected snails during December through February than at any other time of the year. In selected water sites examined at monthly intervals, mean snail density was higher in rice paddies than in other water contact sites but the latter showed a higher prevalence of infected snails than the former. The overall prevalence of Schistosoma mansoni (24.8%) was significantly higher than that of S. haematobium (22.7%) but the difference in prevalence rates of two species in school children was not statistically significant. The intensity of S. haematobium infection (13.2 means G) was significantly higher than that of S. mansoni (6.3 means G). Mixed infections in school children did not have a significant effect on egg output. The prevalence and intensity of S. haematobium showed a dramatic decline between the age groups 0-15 and 20-50 + years old; the differences between these age groups in S. mansoni infection were unremarkable. In Zota, Jorquelle and Kokoya Districts, prevalence rates of S. haematobium were higher than those of S. mansoni; the reverse was observed in Suakoko and Panta-Kpai Districts but relative prevalence rates varied according to specific locality in each district. A south to north stratification of schistosomal infection prevalence was observed similar to the west to east gradient reported by Saladin et al. (1980). New rice paddies developed during the three year operational period of the BCADP contained little or no vector snails and schistosomal infections in farm families of these paddies reflected the characteristic of the disease in corresponding localities.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗

Modes of entry of the first-stage larvae of Elaphostrongylus cervi (Nematoda: Protostrongylidae) into pulmonate snails Arianta arbustorum and Helix pomatia.

Experimental infection of the pulmonate snails Arianta arbustorum L. and Helix pomatia L. with first-stage larvae of protostrongylid nematode Elaphostrongylus cervi Cameron, 1931 was performed in order to determine modes of larval entry into the body of the snail intermediate host. Groups by four individuals of both snail species were examined histologically 30, 60, 90, and 120 minutes after the beginning of exposure and 1, 2, 4, and 7 days post infection. All 64 snails examined were found to be successfully infected. The superficial furrows of the sole were recognized as the most important site of larval entry into the snail organism. Larval penetration was observed to be accompanied by destruction of the superficial epithelium. The number of larvae found in the subepithelial connective tissue of the headfoot was significantly higher than that found in other tissues and organs. Larval counts in individual parts of the body of snails examined from 0 to 7 days p.i. did not fluctuate significantly. The present results indicate that only those protostrongylid larvae which actively penetrated the superficial epithelium of the snail sole play an important role in the life cycle.

Animals↗

Male characteristics on female mud snails caused by antifouling bottom paints.

This study continues an investigation of an anatomical abnormality, named 'imposex', which consists of a superimposition of male characteristics on to a functionally normal female reproductive anatomy of the dioecious snail Nassarius obsoletus Say. Imposex is prevalent in natural populations living near yacht basins and rarely found distant from them. In the current study caged snails were transferred between a yacht basin and a distant 'clean' locality where the natural population of snails was normal. Imposex was induced in some normal snails kept at the marina and suppressed, but not lost in abnormal snails kept at the clean locality. A similar positive result was obtained in the laboratory by exposing normal snails to organotin-containing antifouling paints and abnormal snails to clean sea water. Results were negative in parallel tests of various marina-associated materials which did not contain organotin. The laboratory studies have thus identified a causative factor for the anatomical abnormalities common near yacht basins in the natural environment. They also provide a rare, if not unique, example of a chemical agent which causes the appearance of superfluous anatomical features in an animal.

Animals↗

Antipredatory behavior as an index of heavy-metal pollution? A test using snails and caddisflies.

The loss of behaviors that organisms use to avoid predation may serve as a sensitive indicator of pollution. We tested the hypothesis that a correlation exists in the field between heavy metal levels and antipredator behaviors. We examined the antipredator behavior of aquatic caddisfly larvae and snails at sites in the Coeur d'Alene basin of Northern Idaho which varied in their levels of heavy metals. We tested the antipredator response of Physella columbiana snails at 10 polluted lakes downstream from the Bunker Hill Environmental Protection Agency (EPA) Superfund cleanup site. We then compared their behavior to snails at 14 reference lakes. We placed the snails in a plastic testing apparatus, exposed them to an extract of crushed snail, and then monitored their movements to a normally preferred shaded area. We also tested the behavior of caddisfly larvae from 36 sites from a total of 6 streams/rivers adjacent to the Superfund site. Sites were located upstream and downstream of abandoned mines. We located active larvae of four genera, simulated predation by grasping the animals between thumb and forefinger (the larvae respond to being grasped by withdrawing into their case), lifted them from the water for 3 s, and then placed them in an adjacent, slower section of the stream. We then recorded how long it took each larvae to partially emerge from its case and attempt to move away. Unlike reference site snails, snails from heavy metal-polluted environments failed to exhibit antipredator behaviors in response to crushed conspecifics. These results are consistent with previous laboratory studies. We found no effect of heavy metals on the antipredatory behavior of caddisfly larvae.

Animals↗

Expression of E-cadherin and its repressor snail in placental tissue of normal, preeclamptic and HELLP pregnancies.

Incomplete invasion of extravillous trophoblasts (EVT) is thought to be associated with complications of pregnancy. Snail, a zinc-finger transcription factor represses the transcription of the cell adhesion molecule E-cadherin. The aim of this study was to investigate the expression of E-cadherin and Snail in placental tissue with preeclampsia or HELLP. Placental tissues were obtained from five patients with HELLP syndrome, seven patients with preeclampsia and seven patients after a normal term birth and analysed for Snail and E-cadherin immunoreactivity with specific monoclonal antibodies. Immunohistochemical staining of the placental tissue was analysed using an immunoreactivity score for the evaluation of staining intensity. In preeclamptic EVT, Snail immunoreactivity showed a significant 1.7-fold increase, accompanied by a significant 1.9-fold reduction of E-cadherin immunoreactivity. A 1.7-fold increase of Snail and in parallel a 1.3-fold reduction of E-cadherin was observed in EVT of HELLP placentas although without statistical significance. Loss of E-cadherin can be observed during epithelial-mesenchymal transition (EMT), a key process in normal development and trophoblast differentiation. Snail represses the transcription of E-cadherin triggering a complete EMT. Results obtained in this study showed changes of Snail and E-cadherin immunoreactivity in preeclamptic placentas that could be accompanied with an altered EMT in trophoblasts.

Cadherins↗

Snail regulates p21(WAF/CIP1) expression in cooperation with E2A and Twist.

Snail, a zinc-finger transcriptional repressor, is essential for mesoderm and neural crest cell formation and epithelial-mesenchymal transition. The basic helix-loop-helix transcription factors E2A and Twist have been linked with Snail during embryonic development. In this study, we examined the role of Snail in cellular differentiation through regulation of p21(WAF/CIP1) expression. A reporter assay with the p21 promoter demonstrated that Snail inhibited expression of p21 induced by E2A. Co-expression of Snail with Twist showed additive inhibitory effects. Deletion mutants of the p21 promoter revealed that sequences between -270 and -264, which formed a complex with unidentified nuclear factor(s), were critical for E2A and Snail function. The E2A-dependent expression of the endogenous p21 gene was also inhibited by Snail.

Basic Helix-Loop-Helix Proteins↗

Effects of trematode double infection on the shell size and distribution of snail hosts.

Infection with larval trematodes sometimes alters the phenotypes of their snail hosts. While some trematode species have distinct effects on host phenotypes, it is still unclear how snail phenotypes are altered when they are parasitized with multiple trematode species. Here, we report that double infection with trematode species averages the effects of parasitic alteration on host phenotype. We found that snail hosts Batillaria attramentaria (Batillariidae) infected with Cercaria batillariae (Heterophyidae) have abnormally large shells and distribute in lower areas of the intertidal zone. Snails with another dominant trematode species, the renicolid cercaria I (Renicolidae), have slightly larger shells and distribute in upper areas of the intertidal zone. A number of double infections with both trematodes was observed in this study. Snails infected with both trematode species exhibited an intermediate size and inhabited a depth between those of snails solely infected with either trematode species, suggesting that the two trematodes simultaneously affected the snail phenotypes. Because altered host phenotypes are frequently beneficial to parasites, two trematode species may compete for successful transmission through alteration of host phenotypes.

Animals↗