Toxicity of cadmium and zinc mixtures to the decaudized cercarial life span of Diplostomum spathaceum

The effects of cadmium and zinc mixtures at concentrations ranging from 0.1 to 10,000 microg l(-1) on the life-span of decaudized cercarial bodies (cercariae that have shed their tails) of Diplostomum spathaceum (Trematoda: Diplostomatidae) was investigated. Cercariae were exposed to metal mixtures of equal and unequal concentrations, and a low-dose pre-treatment followed by a high-dose exposure mixtures. Metal mixtures demonstrated variable effects on decaudized cercariae either by increasing or reducing their life-span compared to single metal exposures dependent on concentration and the type of mixed metal treatment. Prolonged exposure to equal metal mixtures at low concentrations (0.1-100 microg l(-1)) resulted in a reduction in the life-span of decaudized cercariae at 0.1 and 100 microg l(-1) in those individuals decaudized during the initial 24 h exposure period compared with those decaudized during the final 24 h period of cercarial survival, whilst in controls there was no significant life-span change between the two time periods. Decaudized cercariae which were exposed to low concentrations (0.1-100 microg l(-1)) of equal metal mixtures were also evaluated for their role as an indicator of larval 'fitness' for migrating through the tissues of their target fish host for those individuals decaudized during the initial 24 h exposure period, and demonstrated only a limited change in their life-span compared to control and single metal exposures. The importance of metal mixtures in parasite establishment in the fish host is discussed.     

Cadmium toxicity and snail-digenean interactions in a population of Lymnaea spp

The toxicity of cadmium to a population of Lymnaea peregra and L. stagnalis naturally infected with a range of digeneans and collected from a number of sites in the lower Thames Valley, UK was investigated. Lymnaeid snails were exposed to 100 microg l-1 cadmium and the effects on host survival and emergence of cercariae recorded. Overwintered L. peregra, but not L. stagnalis, showed significantly reduced survival compared to seasonally infected snails, i.e. snails which have acquired an infection during the spring or summer. A significant increase in survival with increasing snail size was demonstrated for L. stagnalis and for seasonally infected L. peregra only. Only L. stagnalis infected with Diplostomum spathaceum and L. peregra infected individually with D. spathaceum, Sanguinicola inermis, Echinoparyphium recurvatum and Notocotylus attenuatus demonstrated a significantly reduced survival compared to laboratory-bred controls. The exposure of L. stagnalis to cadmium resulted in a significant reduction in the emergence of D. spathaceum over a 5-day period but cadmium-exposed L. peregra showed no difference in the emergence of E. recurvatum cercariae over a 3-day exposure period. The mechanisms and importance of metal toxicity to snail-digenean interactions are discussed.     

Toxicity of cadmium and zinc to Diplostomum spathaceum (Trematoda: Diplostomidae) cercarial survival

The toxicity of cadmium and zinc at concentrations ranging from 0.1 to 10000 microg/l was investigated with cercariae of Diplostomum spathaceum at three temperatures (12, 20, 25 degrees C) and three levels of water hardness (distilled water, soft water, hard water). Under most environmental conditions survival of cercariae was reduced by increasing metal concentration. Increasing water hardness and decreasing water temperature caused an increase in the survival of both control and metal exposed cercariae. However, with decreasing temperature increased survival of metal-exposed cercariae above the control occurred at a number of low metal concentrations. Differences in the relative toxicity of cadmium and zinc to cercariae was dependent on the environmental conditions of exposure. The two heavy metals demonstrated little toxicity to cercariae during the period of maximum cercarial infectivity (0-5 h).The mechanisms of metal toxicity and their effects on cercarial survival are discussed.     

Toxic effects of cadmium and zinc on the transmission of Echinoparyphium recurvatum cercariae

The toxicity of cadmium, zinc and Cd/Zn mixtures to the transmission of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae into the snail second intermediate hosts was investigated at concentrations ranging from 100 microg l-1 to 10 000 microg l-1 in both soft and hard water. A differential response in the infectivity of metal-exposed cercariae into Lymnaea peregra and Physa fontinalis was demonstrated which was dependent on the snail species being infected. Exposure of L. peregra, P. fontinalis, and L. stagnalis to heavy metals caused a differing susceptibility to E. recurvatum cercariae depending on the snail species being exposed. The mechanism and effects of metal toxicity, together with the importance of the parasite/host strain on cercarial transmission are discussed.     

Effects of cadmium and zinc toxicity on orientation behaviour of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae

The effects of cadmium and zinc toxicity on orientation behaviour (photo- and geo-taxis) of Echinoparyphium recurvatum cercariae was investigated at concentrations ranging from 10 to 1000 microg l(-1). Exposure to the toxicants at all metal concentrations caused a change in orientation to negative phototaxis and positive geotaxis during the submaximal dispersal phase (0.5 h cercarial age). Autometallography staining of cercariae exposed to 1000 microg l(-1) cadmium or zinc showed selective binding of heavy metals to tegumental surface sites associated with sensory receptors. The significance to parasite transmission of changes in cercarial orientation behaviour in metal polluted environments is discussed.     

Rhythmicity in the emergence of Cercaria tondoni sp., a gymnocephalus cercaria from Lymnea stagnalis (Linnaeus, 1758)

Emerging pattern of Cercaria tondoni sp., a distome (possessing a ventral sucker lying some distance from the posterior end and approximately at anterior third of the body) belonging to gymnocephalus group (having an oval body and tail devoid of stylets or spines), shows a daily rhythm, being greater during the light phase (0630:1830?h) – similar to that found in a freshwater snail Lymnea stagnalis, Linnaeus, 1758. The peak time of emergence 1230–1430?h has been measured, and there is a high risk for invertebrate or vertebrate hosts and other diurnal animals to acquire infection during this period. However 20% of total shedding in a population of cercariae has also been recorded during the dark phase (0630–1830?h) but there was no significant rhythmicity within this period. An attempt has been made to record cercarial emergence throughout the year, which is of great importance for human welfare. It was observed that the months of April, May, and June are those where frequency of emergence was highest. Implications of this rhythmicity are considered.     

Diplostomum spathaceum (Rud. 1819): effect of miracidial age and lifespan on miracidial infectivity

Experiments were performed to study the effect of age and lifespan on the infection success of Diplostomum spathaceum miracidia. Specimens of Lymnaea peregra and L. stagnalis were individually exposed to single miracidia of D. spathaceum of different ages under conditions of constant illumination at 20 degrees C. The life-span of the free-swimming miracidium of D. spathaceum is 24 h at 20 degrees C. It was observed that as the miracidium ages, its ability to successfully infect snails declines rapidly.     

Putative immune response of rainbow trout, Salmo gairdneri, to Diplostomum spathaceum infections

The immune response of rainbow trout to infection with the cercariae of the digenean parasite Diplostomum spathaceum was investigated. Rainbow trout infected with cercariae at weekly intervals, or injected with a suspension of dead cercariae, did not produce a specific humoral response against D. spathaceum cercariae, as tested using immunoperoxidase, agglutination and cytotoxicity assays. However, a significant difference occurred in the rate of infection of rainbow trout given weekly exposure to cercariae of D. spathaceum in winter and summer. Rainbow trout injected with a suspension of dead cercariae acquired significantly fewer metacercariae than controls when exposed to a challenge infection. This suggested a specific immune response by the host and is the first example of a reduction in the infection rate of rainbow trout immunized against a digenean parasite, when exposed to a challenge infection.     

Diplostomum spathaceum cercariae respond to a unique profile of cues during recognition of their fish host

During its normal life cycle, Diplostomum spathaceum cercariae attach to and invade fish intermediate hosts. They are also known to attach to various other aquatic animals in response to water currents, touch and carbon dioxide. The purpose of this study was to identify the specific stimuli used by D. spathaceum cercariae to recognise the appropriate fish host. We characterised the host cues which stimulate them to remain on the host (enduring contact) and to penetrate the skin. Cercariae were exposed to animal skin tissues and fish skin surface mucus, their extracts and chemical modifications integrated into agar or offered via membrane filters. Enduring contact was stimulated by hydrophilic extracts Mr<3kDa, which were sensitive to oxidation of carbohydrates. The stimulating cues are probably small molecular carbohydrates, as monosaccharides stimulated enduring contacts, but amino acids, urea, electrolytes and peptides did not. Penetration was stimulated by hydrophilic macromolecules, Mr>30kDa, and by lipids. The hydrophilic stimuli were protease resistant and precipitable with Alcian blue and they were sensitive to alkaline cleavage, to digestion with lysozyme and neuraminidase as well as to oxidation of sialic acids. They were considered to be glycoproteins with O-glycosidically linked carbohydrate chains and bound sialic acids as signal structures. The lipophilic penetration stimuli were contained exclusively in the fatty acid fractions, and the stimulating characteristics of these fatty acids resembled the stimulating penetrations in other cercarial species. Diplostomum spathaceum cercariae respond to a unique profile of cues in their sequence of host-recognition phases. These cues differ from those used in other fish parasites studied to date and underline the diversity of fish recognition strategies.     

Survival and infectivity of Hypoderaeum conoideum and Euparyphium albuferensis cercariae under laboratory conditions.

The survival characteristics of the cercariae of Hypoderaeum conoideum and Euparyphium albuferensis (Trematoda: Echinostomatidae) at 20 degrees C and 30 degrees C are described, and the age dependency of their infectivity at 20 degrees C is studied to determine their respective transmission efficiencies. Cercarial survival was found to be age-dependent and was higher at 20 degrees C. For both cercariae, the maximum life-span was 26 h at 20 degrees C and 16 h at 30 degrees C, and their respective times to 50% mortality were similar at each temperature. Both cercariae seem to be well adapted to transmission in their natural habitat, though cercarial infectivity of H. conoideum was higher than that of E. albuferensis, this being correlated with their prevalences in nature. The age-dependency of cercarial survival may be related to steadily diminishing endogenous energy levels, though the delay in attaining maximum infectivity suggests that other factors not related to energy considerations are involved in the delimitation of the cercarial infective period. However, this latter observation may constitute an adaptative mechanism allowing cercarial dissemination.     

Effects of snail size on encystment of Echinostoma caproni in juvenile Biomphalaria glabrata (NMRI strain) and observations on the survival of infected snails

The effects of snail size on encystment of Echinostoma caproni cercariae in neonatal and juvenile Biomphalaria glabrata (NMRI strain) snails were studied. Encystment in neonatal (0.7-1.1 mm shell diameter) and juvenile (2-3 mm shell diameter) snails was compared 24 h post-infection (PI) following individual exposure of snails of each size to 1, 5, 10, 25 and 50 cercariae. Significantly more cysts were recovered from juveniles exposed to 1, 5, 10 and 50 cercariae than from neonatals with comparable exposure. Size of B. glabrata was a major factor in determining cyst burden in this planorbid. Survival of infected versus uninfected neonatals and juveniles was also examined for 7 days. Neonatals exposed to 10 cercariae showed a significant decrease in survival at 3, 6 and 7 days PI when compared to the uninfected controls. There was no significant decrease in the survival of juveniles exposed to 10 cercariae compared to uninfected controls at any time point. Snail size was a factor in mortality associated with echinostome cercarial penetration and encystment.     

Circadian Pattern of Cercarial Emergence in Schistosoma Mansoni(Platyhelminthes: Digenea) from Isolated Biomphalaria Glabrata

The present work aimed to compare the acrophases (peak hours) of emergence of Schistosoma mansoni cercariae among isolated individuals of the snail Biomphalaria glabrata. Laboratory stocks of melanic B. glabrata from the same biotope as the S. mansoni strain (Belo Horizonte, Minas Ger-ais) were used. Twenty-two snails individually exposed to five miracidia were tested. Chronobiological trials were performed outdoors after an acclimation period of at least a week. Three groups of snails were tested between November 1989 and April 1991. Cercarial emergence from individual isolated snails was quantified every 3 h for 3 consecutive days. In all trials, most cercariae were found to emerge during daytime (94.9%). Time series and chronograms showed recurrent peaks during the daytime. The periodogram suggested that 24 h was the period that best fitted cercarial emergence data in 90.9% of the snails. The single cosinor analysis confirmed 24-h rhythms in 95.5% of the snails. Acrophases of cercarial emergence among individual snails occurred between 14:15 and 17:02. They did not differ significantly. The population cosinor analysis indicated greater homogeneity in the 24-h rhythms of cercarial emergence than in the snail groups of each chronobiological trial. Acrophases of cercarial emergence occurred between 14:53 and 15:27 and did not differ significantly among all trials. Data from the three trials were pooled and analyzed using the population cosinor. This statistical method indicated a homogeneity in the 24-h rhythms of cercarial emergence from all snails, with acrophase occurring around 15:00. Results showed that the acrophases of cercarial emergence of S. mansoni are similar among isolated B. glubrura specimens. Data support the hypothesis of a “gate” rhythm in the dynamics of cercarial production and emergence. It is suggested that the adaptive importance of the “gate” mechanism is associated with the concentration of cercariae in the water at times when the vertebrate is present, optimizing the contact between the parasite and the host. The emergence of some cercariae at night (5.1% of the total number of emerged cercariae) suggests a flexible “gate” that could be associated with a residual light effect or with experimental procedures in the laboratory.     

Phenotypic variation in infectivity of Diplostomum spathaceum cercariae within a population

The present study examined phenotypic variation in infectivity of Diplostomum spathaceum (Trematoda) cercariae within a natural population. Twelve infected Lymnaea stagnalis were collected from the field, and the infectivity of cercariae from individual snails was assessed under constant laboratory conditions. At a water temperature of 16.3 C, the mean infectivity of cercariae from the snails varied between 55.5% and 87.5%. Depending on the source of variation, this may have important ecological and evolutionary implications for both natural parasite populations and those occurring in aquaculture.     

Complement-mediated tail degradation of Neodiplostomum seoulense cercariae

The furcocercus cercariae of Neodiplostomum seoulense (Digenea: Neodiplostomidae) penetrate the skins of tadpoles and shed their tails. The speculated mechanism of this tail loss was physical efforts required to produce a vigorous zigzag motion during skin penetration; no other mechanism has been proposed. We examined the relationship between the host serum and cercarial tail loss. Cercariae of N. seoulense were collected from experimentally infected Segmentina hemisphaerula, and lots of 300 cercariae were cultured in medium 199 contained several types of sera. Cercarial tail degradation was induced in all media, but all the cercariae cultured except those cultured in media containing fetal bovine serum (FBS) died within 48 hr. After 72 hr cultivation in media containing FBS, cercarial tail degradation was induced in 67.0%; in continuous cultivation 13.3% of larvae survived for 7 days. Tail degradation did not occur in the absence of serum and when serum was heat inactivated at 56 degrees Celsius for 30 min. The addition of 20 mM ethylenediaminetetraacetic acid (EDTA) blocked cercarial tail degradation completely. Moreover, the addition of 20 mM MgCl2 restored tail degradation blocked by EDTA. These results suggest that the alternative complement pathway is related with the N. seoulense cercarial tail degradation induced by serum.     

Bioelectronic monitoring of parasite-induced stress in brown trout and roach

A non-contact bioelectronic monitoring system was used to record changes in heart rate, ventilation rate and swimming activity in brown trout Salmo trutta and roach Rutilus rutilus , following exposure to two species of cercariae of digenean trematodes: Diplostomum pseudospathaceum which is a common parasite in the lens of fishes and xiphidiocercariae of Plagiorchis elegans , a parasite of anatid birds, both of which have the snail Lymnaea stagnalis as their first intermediate host. Swimming activity increased significantly in roach exposed to Diplostomum cercariae at densities as low as 3.8l⁻¹ and remained elevated for 24–36 h post exposure. Brown trout showed no response when exposed to low densities of Diplostomum (2.9 and 5.7 cercariae 1⁻¹) but there was a significant response at higher cercarial exposure densities (>501⁻¹). Total activity of brown trout increased immediately, peaking after 2 h and returning to pre-exposure levels within 5–6 h. There was a simultaneous increase in heart and ventilation rates which both declined following the reduction in activity of the fish. Heart rate then increased for a second time to a maximum level at 14–16 h and thereafter declined to reach a steady state at 3 days post-exposure. During this period there was no significant change in total activity. The qualitative nature of the heart response was identical in five infected brown trout although the magnitude of the response varied widely among fish. Exposure to cercariae of P. elegans elicited no response in either fish species. (c) 1996 The Fisheries Society of the British Isles     

Cercarial tail loss in Echinostoma caproni: the influence of in vivo encystment and copper sulphate

Echinostoma caproni tail loss was studied in vitro in the presence of the toxicant copper sulphate (CuSO4) in concentrations ranging from 1 to 10 000 mg l(-1) in standardized artificial spring water (pH 7.4, osmolarity 34 mOsm kg(-1) H2O, Ca(2+) 20 mg l(-1)) at 23 degrees C. Tail loss was also studied in the absence of toxicants during in vivo encystment of the cercariae in juvenile Biomphalaria glabrata. As the concentration of CuSO4 increased, the percentage of cercarial tail loss increased. By 2 h in 10 000 mg l(-1), 1000 mg l(-1) and 100 mg l(-1) CuSO4, 50%, 23% and 13%, respectively, of the cercariae had lost their tails. In the in vivo studies, by 1 h PI, 59+/-5% of cercariae had lost their tails and only 4+/-1% of the cercariae were actively swimming in the multi-well dishes. At 3 h PI, 72+/-3% of the cercariae began to form cysts within the snails.     

The presence of predators modifies the larval development of Fasciola hepatica in surviving Lymnaea truncatula

Experimental infections of Lymnaea truncatula with Fasciola hepatica were performed to study the consequences of the presence of predators (sciomyzid larvae or zonitid snails) on the characteristics of larval F. hepatica development in surviving snails. Controls consisted of infected snails that were not subjected to predators. Compared to controls, the survival rate at day 30 post-exposure, the duration of cercarial shedding, and the number of cercariae shed by surviving snails were significantly lower when predators were present in snail breeding boxes, whatever the type of predator used. In contrast, the prevalences of Fasciola infections in snails, and the length of time between exposure and the onset of cercarial shedding showed no significant variation. The progressive development of a stress reaction in surviving snails against predators during the first 30 days of experimental exposure to F. hepatica would influence snail survival during the cercarial shedding period and, consequently, the number of cercariae shed by the snails.     

Fasciola gigantica: Cercarial shedding pattern from Lymnaea natalensis after long-term exposure to cadmium at different temperatures

Little is known about the response of Fasciola spp., inside its snail host to pollutant toxicity. The effect of long-term exposure of Lymnaea natalensis to 0.1 μg/l, 10 μg/l, or 100 μg/l cadmium (Cd) on the infection rates with F. gigantica, percentage of cercariae-shedding snails and cercariae shedding pattern was investigated. The snails were exposed to Cd for 7 days either pre-infection with single Fasciola miracidium or during the late pre-patent period of the infection. The possible interaction between metal exposure and acclimatization temperature was also studied in three ranges; 16-18 °C, 23-25 °C and 30-32 °C. Results clearly showed that host exposure to 100 μg/l Cd significantly affected the pattern of Fasciola development inside its snail host, and that the acclimatization temperature was a key factor affecting the role played by Cd. Pre-infection exposure to Cd caused a significant reduction in the infection rates where the effect was temperature-dependent. Post-infection exposure to Cd significantly increased the percentage of cercariae-shedding snails; the effect was temperature-independent. Interestingly, post-infection exposure significantly altered the differential cercarial output pattern, while no clear impact on the total was found. The alteration in the differential cercarial output was represented as a significantly higher proportion of floating metacercaria as compared to fixed ones and also higher proportion of dead cercariae which may directly reduce the transmission of Fasciola to the final host. The mechanisms of cadmium impact are briefly discussed.     

Patterns and processes of larval emergence in an estuarine parasite system

Trematode parasites in intertidal estuaries experience constantly varying conditions, with the presence or absence of water potentially limiting larval transport between hosts. Given the short life spans (< or =24 h) of cercariae, emergence timing should be optimized to enhance the probability of successful transmission. In the present study, field measurements and laboratory experiments identified processes that regulate the emergence of cercariae from their first intermediate snail hosts in an intertidal marsh. Larvae emerged over species-specific temperature ranges, exclusively during daylight hours, and only when snails were submerged. The three factors operate over different temporal scales: temperature monthly, light diurnally (24-h period), and water depth tidally (12-h period). Each stimulus creates a necessary condition for the next, forming a hierarchy of environmental cues. Emergence as the tide floods would favor transport within the estuary, and light may trigger direct (downward or upward) swimming toward host habitats. Abbreviated dispersal would retain asexually reproduced cercariae within the marsh, and local mixing would diversify the gene pool of larvae encysting on subsequent hosts. In contrast to the timing of cercarial release, emergence duration was under endogenous control. Duration of emergence decreased from sunrise to sunset, perhaps in response to the diminishing lighted interval as the day progresses. Circadian rhythms that control cercarial emergence of freshwater species (including schistosomes) are often set by the activity patterns of subsequent hosts. In this estuary, however, the synchronizing agent is the tides. Together, exogenous and endogenous factors control emergence of trematode cercariae, mitigating the vagaries of an intertidal environment.     

Delayed tail loss during the invasion of mouse skin by cercariae of Schistosoma japonicum

A traditional assumption is that schistosome cercariae lose their tails at the onset of penetration. It has, however, recently been demonstrated that, for Schistosoma mansoni, cercarial tails were not invariably being shed as penetration took place and a high proportion of tails entered human skin under experimental conditions. This phenomenon was termed delayed tail loss (DTL). In this paper, we report that DTL also happens with S. japonicum cercariae during penetration of mouse skin. It occurred at all cercarial densities tested, from as few as 10 cercariae/2·25 cm(2) of mouse skin up to 200 cercariae. Furthermore, it was demonstrated that there was a density-dependent increase in DTL as cercarial densities increased. No such density-dependent enhancement was shown for percentage attachment over the same cercarial density range.