Cercarial Release Research Articles

Human-water interactions associated to cercarial emergence pattern and their influences on urinary schistosomiasis transmission in two endemic areas in Mali

BackgroundMali is known to be a schistosomiasis-endemic country with a limited supply of clean water. This has forced many communities to rely on open freshwater bodies for many human-water contact (HWC) activities. However, the relationship between contact with these water systems and the level of schistosome infection is currently receiving limited attention. This study assessed human-water interactions including cercarial emergence pattern and their influences on urinary schistosomiasis transmission in two communities in the Kayes district of Mali.MethodsWe carried out a parasitological study first in children in September 2021, then a cross-sectional study of quantitative observations of human-water contact activities in the population, and finally a study of snail infectivity at contact points in September 2022. The study took place in two communities, Fangouné Bamanan and Diakalèl in the Kayes region of western Mali. The chronobiological study focused on cercarial release from naturally infected snails. Released cercariae were molecularly genotyped by targeting the cox1 region, and the ITS and 18S ribosmal DNA gene (18S rDNA) regions of the DNA. Links between sociodemographic parameters, human water-contact points and hematuria were established using multivariate statistical analysis or the logistic regression model.ResultsThe main factor predisposing the 97 participants to water contact was domestic activity (62.9%). Of the 378 snails collected at 14 sampling sites, 27 (7.1%) excreted schistosome cercariae, with 15.0% (19/126) at Fangouné Bamanan and 3.3% (8/252) at Diakalel. The release of Schistosoma cercariae shows three different patterns in Fangouné Bamanan: (i) an early release peak (6:00–8:00 AM), (ii) a mid-day release peak (10:00 AM–12:00 PM) and (iii) a double peak: (6:00–8:00 AM) and (6:00–8:00 PM) cercariae release; and two release patterns in Diakalel: early release (6:00–8:00 AM) and (ii) mid-day release (12:00–2:00 PM). All cercariae released during early diurnal (6:00–8:00 AM) or nocturnal emission patterns (6:00–8:00 PM) were hybrids parasite having an cox1 S. bovis or S. curassoni associated with an ITS and 18S rDNA of S. haematobium while the cercariae released during diurnal, or mid-day patterns (8:00 AM–6:00 PM) were pure S. haematobium.ConclusionsOur study showed that domestic activity is the main source of exposure in the Kayes region. Two and three cercariae emission patterns were observed at Diakalel and Fangouné Bamanan respectively. These results suggest that the parasite adapts to the human-water contact period in order to increase its infectivity.Graphical

Abundance, Distribution, and Diversity of Freshwater Snail and Prevalences of Their Infection by Cercaria of Fasciola gigantica and Schistosoma spp at Mayo-Vreck River, Far North Region of Cameroon.

Malacological and parasitological studies were conducted from April 2020 to March 2021 to determine the abundance and distribution of molluscs and cercariae of Schistosoma spp and Fasciola gigantica. Collected molluscs are exposed to strong light to induce cercarial release. Mollusc densities were higher at station 1 (Gamak) than in station 8 (Patakai), with Bellamya unicolor and Biomphalaria pfeifferi more abundant and Bulinus truncatus, B. tropicus, and B. globosus less abundant. The overall prevalence of cercariae (19.87%) is higher in station 3 (Yaye orchard), station 9 (Gougni), station 4 (Madiogo), station 5 (Madiogo pasture), and station 6 (Ziam 3). It varies significantly between 15.76% in station 8 and 25.77% in station 3, between 8.48% in B. truncatus and 25.53% in B. globosus, and between 19.27% for cercariae of Schistosoma spp and 21.60% for those of F. gigantica. Cercarial emissions in L. natalensis and B. pfeifferi were higher in hot and cold dry seasons; on the other hand, cercarial emissions in B. globosus were higher in hot dry seasons (31.48%) and rainy seasons (23.38%). Emissions of cercariae from S. haematobium are related to areas of human activity and defecation, while those of F. gigantica in L. natalensis, Schistosoma haematobium in B. tropicus, and S. mansoni in B. pfeifferi are related to grazing areas. Mayo-Vreck is a site that favors the endemicity of fascioliasis and human schistosomiasis.

Cercariae of a Bird Schistosome Follow a Similar Emergence Pattern under Different Subarctic Conditions: First Experimental Study.

The emergence of cercariae from infected mollusks is considered one of the most important adaptive strategies for maintaining the trematode life cycle. Short transmission opportunities of cercariae are often compensated by periodic daily rhythms in the cercarial release. However, there are virtually no data on the cercarial emergence of bird schistosomes from freshwater ecosystems in northern latitudes. We investigated the daily cercarial emergence rhythms of the bird schistosome Trichobilharzia sp. “peregra” from the snail host Radix balthica in a subarctic lake under both natural and laboratory seasonal conditions. We demonstrated a circadian rhythm with the highest emergence during the morning hours, being seasonally independent of the photo- and thermo-period regimes of subarctic summer and autumn, as well as relatively high production of cercariae at low temperatures typical of northern environments. These patterns were consistent under both field and laboratory conditions. While light intensity triggered and prolonged cercarial emergence, the temperature had little effect on cercarial rhythms but regulated seasonal output rates. This suggests an adaptive strategy of bird schistosomes to compensate for the narrow transmission window. Our results fill a gap in our knowledge of the transmission dynamics and success of bird schistosomes under high latitude conditions that may serve as a basis for elucidating future potential risks and implementing control measures related to the spread of cercarial dermatitis due to global warming.

Contrasting temperature responses in seasonal timing of cercariae shedding by Rhipidocotyle trematodes.

Global warming is likely to lengthen the seasonal duration of larval release by parasites. We exposed freshwater mussel hosts, Anodonta anatina, from 2 high-latitude populations to high, intermediate and low temperatures throughout the annual cercarial shedding period of the sympatric trematodes Rhipidocotyle fennica and R. campanula, sharing the same transmission pathway. At the individual host level, under warmer conditions, the timing of the cercarial release in both parasite species shifted towards seasonally earlier period while its duration did not change. At the host population level, evidence for the lengthening of larvae shedding period with warming was found for R. fennica. R. campanula started the cercarial release seasonally clearly earlier, and at a lower temperature, than R. fennica. Furthermore, the proportion of mussels shedding cercariae increased, while day-degrees required to start the cercariae shedding decreased in high-temperature treatment in R. fennica. In R. campanula these effects were not found, suggesting that warming can benefit more R. fennica. These results do not completely support the view that climate warming would invariably increase the seasonal duration of larval shedding by parasites, but emphasizes species-specific differences in temperature-dependence and in seasonality of cercarial release.

Species-specific patterns in cercarial emergence of Diplostomum spp. from snails Radix lagotis

The cercarial emergence patterns of three species of Diplostomum (Diplostomum ‘mergi’, Diplostomum spathaceum and Diplostomum parviventosum) parasitizing freshwater first intermediate host Radix lagotis sampled in Most Lake, Czech Republic, were studied under various experimental conditions, i.e. field, laboratory and incubator, and seasons, i.e. spring, summer and autumn. We discovered unexpected daily periodicity-dependent species-specific emergence patterns among the three Diplostomum spp. depending on experimental conditions. At the same time, the intraspecific variation of D. spathaceum cercarial release in response to seasonal conditions was observed. We found that a complex array of mechanisms can affect Diplostomum species-specific patterns in cercarial emergence, of which behavioural characteristics of fish related to reproduction and feeding processes are considered the most important factors. This might represent a specific adaptive evolutionary mechanism to maximise transmission success while avoiding competition for host resources. Our results contribute to a better understanding of ecological and epidemiological aspects with respect to specific adaptive strategies compartmentalised among species of Diplostomum and consequences for infection risk in fish hosts.

Aquatic macrophytes and macroinvertebrate predators affect densities of snail hosts and local production of schistosome cercariae that cause human schistosomiasis.

BackgroundSchistosomiasis is responsible for the second highest burden of disease among neglected tropical diseases globally, with over 90 percent of cases occurring in African regions where drugs to treat the disease are only sporadically available. Additionally, human re-infection after treatment can be a problem where there are high numbers of infected snails in the environment. Recent experiments indicate that aquatic factors, including plants, nutrients, or predators, can influence snail abundance and parasite production within infected snails, both components of human risk. This study investigated how snail host abundance and release of cercariae (the free swimming stage infective to humans) varies at water access sites in an endemic region in Senegal, a setting where human schistosomiasis prevalence is among the highest globally.Methods/Principal findingsWe collected snail intermediate hosts at 15 random points stratified by three habitat types at 36 water access sites, and counted cercarial production by each snail after transfer to the laboratory on the same day. We found that aquatic vegetation was positively associated with per-capita cercarial release by snails, probably because macrophytes harbor periphyton resources that snails feed upon, and well-fed snails tend to produce more parasites. In contrast, the abundance of aquatic macroinvertebrate snail predators was negatively associated with per-capita cercarial release by snails, probably because of several potential sublethal effects on snails or snail infection, despite a positive association between snail predators and total snail numbers at a site, possibly due to shared habitat usage or prey tracking by the predators. Thus, complex bottom-up and top-down ecological effects in this region plausibly influence the snail shedding rate and thus, total local density of schistosome cercariae.Conclusions/SignificanceOur study suggests that aquatic macrophytes and snail predators can influence per-capita cercarial production and total abundance of snails. Thus, snail control efforts might benefit by targeting specific snail habitats where parasite production is greatest. In conclusion, a better understanding of top-down and bottom-up ecological factors that regulate densities of cercarial release by snails, rather than solely snail densities or snail infection prevalence, might facilitate improved schistosomiasis control.

Effect of Temperature on the Release of Proterometra macrostoma (Trematoda: Digenea) Cercariae from Their Snail Intermediate Host, Pleurocera semicarinata (Gastropoda: Pleuroceridae) at North Elkhorn Creek, Kentucky

The effect of temperature on cercarial release has been well documented for several digenean species; however, there have been no prior studies assessing shedding of cercariae from snails infected with the azygiid, Proterometra macrostoma. Snails, Pleurocera semicarinata, were collected at the Lemon's Mill site on North Elkhorn Creek in Scott County, Kentucky during June and July 2016. Snails shedding cercariae (n = 453) were assigned to temperature treatments (10, 13, 15, 18, 20, 25, and 30°C) that were within the range of previously reported seasonal water temperatures at North Elkhorn Creek. Snails were acclimated to experimental temperatures for 3 d. Cercarial counts were then made at 0900 h daily over the next 7 d, and the average 7-d cercarial release was calculated for each snail. The minimum emergence threshold temperature for Pr. macrostoma cercarial release was determined to be approximately 13°C, whereas the minimum development threshold temperature was between 10 and 12°C. Notably, cercarial release increased with temperature up to 25°C, but declined at 30°C. Q10 values calculated from 10°C intervals were greatest at the lowest (13–23°C) temperature interval (Q10 = 16.1), and declined at the higher (15–25 and 20–30°C) intervals (Q10 = 4.3 and Q10 = 1.8, respectively). Snail survivorship within combined replicates ranged from 87.1 to 100% over the 10-d experiment, with the lowest survivorship observed at 30°C. These results are discussed with regard to climate change and global warming.

Cercarial emergence patterns for Opisthorchis viverrini sensu lato infecting Bithynia siamensis goniomphalos from Sakon Nakhon Province, Thailand.

Opisthorchis viverrini sensu lato is a food-borne trematode which is classified as a class 1 carcinogen, with infection potentially leading to cholangiocarcinoma. Snails of the genus Bithynia act as the first intermediate hosts and an amplifying point in the parasite life cycle. In order to investigate seasonal effect on transmission dynamics of O. viverrini in Bithynia siamensis goniomphalos, cercarial emergence and output profiles were monitored at different season. A total of 4533 snails originating from Sakon Nakhon Province, Thailand, collected during the three main seasons, were analyzed for O. viverrini s.l. Emergence of O. viverrini s.l. cercariae from snails was monitored daily from 06:00 to 18:00h for seven consecutive days. The prevalence of infection in the snails was highest in the hot-dry season and declined in the rainy and cool-dry seasons. Peak cercarial emergence occurred between 08:00 and 10:00h during the rainy and cool-dry seasons and between 10:00 and 12:00h during the hot-dry season. The cercarial output was highest in the hot-dry season, similar to a previous study from Lao People's Democratic Republic (PDR). Average cercarial output/snail in Thailand was higher than in Lao PDR. The number of cercariae emerging from the snails was strongly related to snail size, but the relationship between prevalence of infection and snail size differed between seasons. Observed discrepancies in the emergence patterns and per capita cercarial release may reflect differences in environmental, snail, and/or parasite factors particularly biological characteristics between the cryptic species of O. viverrini s.l. and B. s. goniomphalos from Thailand and Lao PDR.

Cercarial shedding of trematodes and their associated snail intermediate hosts in Borno State, Nigeria

ObjectiveTo determine the distribution of cercariae of trematodes and their associated snail intermediate hosts in Borno State, Nigeria. MethodsHandpicking and scooping by the aid of scoop nets were used to collect a total of 1 700 snails. They were examined for cercarial release using emergent method of cercarial release. Results16 (0.94%) of the 1 700 snails were positive for cercariae of trematodes based on morphology and standard measurement. Three species (Bulinus forskalli, Bulinus globosus and Lymnae natalensis) of snails were identified based on shell morphology and standard criteria. Distribution of the infection among the sampled snails showed that, 10 (1.75%) of Bulinus globosus and 6 (0.55%) of Lymnae natalensis released cercariae of Schistosoma haematobium and a distome cercaria of Fasciola spp. respectively. The intensity of infection among the examined snails varied significantly (P<0.05) based on senatorial zones of sampling. ConclusionsThe present study has demonstrated the presence of some important trematodes and snail species that are possible intermediate hosts in Borno State. This study will further provide insight into better understanding of the epidemiology of schistosomiasis caused by Schistosoma haematobium and other trematodes and the role of snails (Bulinus globosus and Lymnae natalensis) as intermediate hosts.

Effect of Short-Term Temperature Change on Cercarial Release by Rhipidocotyle fennica (Trematoda, Bucephalidae) from the Freshwater Bivalve Host, Anodonta anatina

Cercarial release from the first intermediate host is an important stage in the transmission of trematode parasites. Besides long-term (seasonal) temperature fluctuations, short-term temperature changes can also influence cercarial emergence. We tested the response of the bucephalid trematode, Rhipidocotyle fennica (R. fennica), acclimatized to 17 °C, to an abrupt temperature change. As the natural cercarial shedding by this parasite takes place annually during the warmest season, we expected a positive effect of temperature increase. Monitoring during one hour after the transfer from 17 °C to 20 °C revealed a significant increase in R. fennica cercarial release compared to the preceding one hour period. In contrast, cercarial release decreased in clams transferred to 14 °C, while no change was observed in control clams transferred from 17 °C to 17 °C. This shows that the cercarial release by R. fennica is sensitive to short-term temperature change, and, as predicted, responds positively to warming and negatively to cooling. The result emphasizes the importance of (i) temperature on the cercarial production of trematodes, and (ii) the need to carefully control temperature conditions when studying factors influencing the cercarial production of trematodes.

Determination and quantification of Schistosoma mansoni cercarial emergence from Biomphalaria glabrata snails.

Living and fixed samples of Schistosoma mansoni -infected Biomphalaria glabrata snails were used to determine the relative contributions of different snail tissues to cercarial emergence (shedding). Three methods of observations were employed: (1) direct microscopical observations of shedding snails; (2) microscopic analysis of 5 μm serial sections (H&E stained) of actively shedding snails; and (3) scanning electron microscopic (SEM) observations of snails that were fixed while actively shedding. For this investigation, there were advantages and disadvantages to using each method. We confirmed the results of others that there were 3 tissues of the snail that contributed most prominently to cercarial release (mantle collar, pseudobranch, and headfoot). Based on histological analysis of cercarial accumulations in presumed shedding sites in these 3 tissues, 57% of the cercariae could be seen in the mantle collar, 30.6% in the pseudobranch, and 12.5% in the headfoot. Other anterior structures were involved to a much lesser extent. SEM observations clearly showed cercariae emerging either body first, tail first, or likely emerging en masse from blebs, especially from the mantle collar. These studies provide a more quantitative appraisal of the role the different anterior snail tissues play in cercarial emergence.

Encystment and excystment of the paramphistomid trematode Zygocotyle lunata

This study reports further observations on encystment and excystment of the paramphistomid trematode Zygocotyle lunata. Of numerous substrates tested in the laboratory for cercarial encystment, i.e. plastic and glass dishes, Styrofoam™, aluminium foil, snail shells, and cheesecloth, all but cheesecloth allowed for 100% encystment within 4h of cercarial release at 28°C. Numerous physical and chemical factors were tested to kill cysts, and their capacity to do so was determined by the subsequent inability of cysts to excyst within a chemical excystment medium. Vinegar, Italian salad dressing, soy sauce and 10% solutions of sucrose, acetic acid and sodium chloride were the most detrimental. Freezing, boiling and storage of cysts for more than 2 years also killed them. Several marinades such as lemon juice, Worcestershire sauce, and various concentrations of potassium permanganate, did not kill the majority of cysts tested. Since Z. lunata is a potential foodborne trematode for humans and animals, it is important to determine suitable ways to control or kill these cysts.

Dynamics of Diplostomum spathaceum infection in snail hosts at a fish farm

Ecologically sustainable disease prevention in intensive monocultures, such as fish farming, is based on the knowledge of the ecology of parasites and the identification of the key proportion of host populations for parasite life cycles. In this paper, we examined the life cycle dynamics of the pathogenic trematode Diplostomum spathaceum at a fish farm during a period of 1 year, focusing especially on the pattern of infection in the snail host (Lymnaea stagnalis) population, which is the key phase in the parasite life cycle regarding preventative actions. We found that (1) the infection varied seasonally in snails, being highest in late August; (2) the prevalence of infection and the proportion of individuals releasing cercariae were strongly dependent on snail size; (3) the parasite over-wintered in snails as dormant sporocysts and 4) the mortality of infected snails was not likely to differ from uninfected individuals during winter. Furthermore, the seasonality and development of the infection in the snails corresponded to the pattern of infection observed in the tracer rainbow trout caged in the farm area. This pattern of infection also indicated strong spatiality of the infection, probably because of differences in the size of the snail population between the ponds rather than differences in the prevalence of infection. Overall, present results indicate a possibility for late-summer outbreaks of D. spathaceum, but also that snail prevention at farms could be conducted more efficiently by considering both size-dependent infection and the timing of cercarial release.

The effects of temperature change on the infection rate of Biomphalaria glabrata with Schistosoma mansoni

The aim of this study was to investigate the influence of temperature on the development of Schistosoma mansoni infections in Biomphalaria glabrata. The snails were infected at 15, 20, and 30 degrees C, and the cercarial release was analyzed after 30 and 60 days post-infection. Our results showed that a decrease in the temperature has a substantial influence on the development of S. mansoni infection in B. glabrata, with significant differences (p < 0.05) between 15 and 30 degrees C. These data could provide a better understanding of the epidemiological aspects of schistosomiasis.

Emergence of cercariae of Echinostoma caproni and Schistosoma mansoni from Biomphalaria glabrata under different laboratory conditions.

Release of Echinostoma caproni cercariae and Schistosoma mansoni from experimentally infected Biomphalaria glabrata snails maintained under different laboratory conditions was studied. Infected snails were isolated individually for 1 h in Stender dishes containing 5 ml of artificial spring water and the number of cercariae released during this time was recorded. Of numerous conditions tested, the addition of lettuce, the use of water conditioned by B. glabrata snails and a temperature of 35 degrees C significantly increased the release of E. caproni cercariae. A significant increase in cercarial release of S. mansoni was seen only in cultures fed lettuce. A temperature of 12 degrees C caused a significant decrease in cercarial release of both E. caproni and S. mansoni. Increased snail activity associated with feeding behaviour was probably responsible for the enhanced cercarial sheds observed in this study.

Effect of a short period of desiccation during the patent period on cercarial shedding of Schistosoma haematobium from Planorbarius metidjensis

Planorbarius metidjensis infected with Schistosoma haematobium were subjected, from the first day of shedding, to a 5- or 10-day period of desiccation in order to study its effect on the cercarial release occurring after reactivation of snails in water. These results were compared to those of infected controls that were not subjected to desiccation. Compared to controls, the duration of shedding following reactivation was shorter and the total number of cercariae shed by each infected snail was smaller in the 5- and 10-day groups. The rhythm occurring in the daily cercarial production of the 5-day group was shorter (11 days only) at the beginning of the shedding period and lengthened to 17-18 days at the end of the patent period. In the 10-day group, no rhythm was found. These results could be explained by the length of the desiccation period and its consequences on the development of S. haematobiumsporocysts in the body of snails.

More efficient allopatric combinations of Fasciola hepatica and Lymnaea truncatula due to modification of redial development?

Experimental infections of two susceptible French populations of Lymnaea truncatula (Courcelles and Saint Ours) with an allopatric (Fès, Morocco) or a semi-sympatric (Limoges, France) isolate of Fasciola hepatica miracidia were performed to determine the effect of allopatric miracidia on redial and cercarial production. In both populations, cercarial release was significantly greater in allopatric than in semi-sympatric snails. Compared to semi-sympatric snails, the examination of allopatric snails killed from day 14 to day 35 post-exposure demonstrated (1) a significant decrease after day 28 in the number of daughter rediae (R2a group) exiting from the first-appearing mother redia (R1a redia), and (2) the differentiation of numerous daughter rediae (R2b group) in the body of second-appearing mother rediae (R1b group). These experiments demonstrated that the exposure of L. truncatula to an allopatric isolate of miracidia disturbed the usual developmental pattern of redial generations and caused the formation of numerous R2b rediae. The authors hypothesized that the increase in cercarial release noted in allopatric snails would be assured by the rediae from the R2b group after their emergence from the body of R1b mother rediae.

Trematode Accumulation by the Estuarine Gastropod Ilyanassa obsoleta

The accumulation of larval trematodes by Ilyanassa obsoleta (Gastropoda) at 2 estuarine sites in Delaware was studied. Initial infection status of snails was assayed by looking for shed cercariae. Native snails (most already infected) were deployed at sites A and B, and sentinel snails (putatively uninfected) were deployed at site B. All were individually marked and, if found, reassessed for infection after being free 1-3 summers. Himasthla quissetensis, Lepocreadium setiferoides, Zoogonus rubellus, Austrobilharzia variglandis, and Gynaecotyla adunca infections were observed in recovered snails. At site A, in 1993, 62 natives were recovered. Among the 26 initially testing uninfected, 15 had infections at recovery. Of 36 that released cercariae, 26 had the infection initially indicated. Of sentinels released at site B in 1996, 3 of 16 had infections when recollected. One probably was infected before transplant; at least 2 were infected at site B during 1996-1997. Among site B natives, 26 were later examined by dissection in 1996; 22 had the infection status initially revealed. Some site B natives (n = 35) escaped recapture in 1996 but were found in 1997, or 1998, or both, and were reexamined, most by cercarial release. The same cercariae were produced by 30. Among natives (both sites, n = 123), 27.6% exhibited some difference in infection status compared to the initial assay. This probably overestimates changes. Some differences were real but most can be discounted as cases where initial infection status was misrepresented.

Schistosoma mansoni: The Role of Calcium in the Stimulation of Cercarial Proteinase Release

We investigated the role of calcium mobilization in the induction of proteinase release from cercarial preacetabular glands. Proteinase release was measured by the ability of cercariae to break down a 3H-labeled proline extracellular fibroblast matrix and calcium influx was measured using 45Ca2+. The role of calcium in the activation of cercarial proteinase was examined by investigating the effects of calcium addition and removal on linoleate-induced matrix degradation, the ability of various calcium modulators (Verapamil, fendiline, nifedipine, SK-525A, BAY K-8644, Ryanodine, and SK-7171A) to stimulate or inhibit linoleate-induced proteinase release, the ability of calcium modulators directly to induce cercarial proteinase release, and the ability of various stimulants of proteinase release to induce calcium influx or efflux from cercariae. The results of these studies indicate that proteinase release is dependent on external calcium concentration, voltage-operated channels are either nonexistent in cercariae or have a minimal role in overall calcium influx, and that activation of Ca2+ influx can be caused by both free fatty acids and calcium modulators by a hypothesized receptor-operated channel. Although calcium uptake is important in cercarial proteinase release, it is not the only factor involved. Calcium uptake alone does not guarantee that proteinase will be secreted. On the other hand, if Ca2+ influx does not occur, proteinase will not be secreted.

Trematode infections in a gastropod host misrepresented by observing shed cercariae

Cercarial emergence is often used to determine trematode infections in gastropods. Specimens of Ilyanassa obsoleta (Say) ( n = 206) were examined for trematode infections by two methods, cercarial emergence and dissection. Dissection revealed five trematode species in 18 infection combinations. Cercarial emergence failed to reveal many infections and thereby severely underestimated parasitism. Percent failed determinations for each trematode species found were: Himasthla quissetensis (Miller and Northup)= 10.8; Lepocreadium setiferoides (Miller and Northup)= 30.6; Zoogonus rubellus (Olsson)= 43.8; Gynaecotyla adunca (Linton)= 80.2; and Austrobilharzia variglandis (Miller and Northup)= 64.7. Infections in snails harboring two and three trematode species were misrepresented in 80 and 93% of cases, respectively. Overall, cercarial release wrongly determined parasitism in 59.1% of infected snails examined. Thus, as a method, cercarial emergence severely underestimates parasite prevalence and overlooks many species that are present.