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In a cultured pikeperch (Sander lucioperca) stock the monopisthocotylean monogenean gill parasite Ancyrocephalus paradoxus caused heavy infection and mortalities. The gills of the affected fish specimens were infected by 50 to 800 monogenean parasites. Severe pathological changes were found in areas where the worms attached to the gills. At the attachment sites the haptoral discs of the worms formed a deep depression in the epithelium of the filaments, and the anchors pierced into and fixed themselves to the connective tissue of the cartilaginous gill rays. At these attachment sites red blood cells released from injured capillaries were found among the damaged epithelial cells. Around the hooks, anchors and body sections coming into contact with the gill filaments a proliferative tissue developed in which only a remnant of the damaged lamellae was found. Due to the damage caused by the worms the tips of the heavily infected gill filaments fused, formed clubs and were composed of epitheloid-type regeneration tissue lacking respiratory lamellae. In the basal parts of the filaments, where most of the worms attached to the gill, only denuded filaments deprived of lamellae were observed among the cross-sectioned worms in histological sections.

Excessive fertilisation is one of the most pernicious forms of global change resulting in eutrophication. It has major implications for disease control and the conservation of biodiversity. Yet, the direct link between nutrient enrichment and disease remains largely unexplored. Here, we present the first experimental evidence that chronic nitrate enrichment decreases severity and induces protection against an infectious disease. Specifically, this study shows that nitrate concentrations ranging between 50 and 250mgNO3(-)/l reduce Gyrodactylus turnbulli infection intensity in two populations of Trinidadian guppies Poecilia reticulata, and that the highest nitrate concentration can even clean the parasites from the fish. This added to the fact that host nitrate pre-exposure altered the fish epidermal structure and reduced parasite intensity, suggests that nitrate protected the host against the disease. Nitrate treatments also caused fish mortality. As we used ecologically-relevant nitrate concentrations, and guppies are top-consumers widely used for mosquito bio-control in tropical and often nutrient-enriched waters, our results can have major ecological and social implications. In conclusion, this study advocates reducing nitrate level including the legislative threshold to protect the aquatic biota, even though this may control an ectoparasitic disease., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The impact of widespread and common environmental factors, such as chemical contaminants, on infectious disease risk in amphibians is particularly important because both chemical contaminants and infectious disease have been implicated in worldwide amphibian declines. Here we report on the lone and combined effects of exposure to parasitic cercariae (larval stage) of the digenetic trematode, Acanthostomum burminis, and four commonly used pesticides (insecticides: chlorpyrifos, dimethoate; herbicides: glyphosate, propanil) at ecologically relevant concentrations on the survival, growth, and development of the common hourglass tree frog, Polypedates cruciger Blyth 1852. There was no evidence of any pesticide-induced mortality on cercariae because all the cercariae successfully penetrated each tadpole host regardless of pesticide treatment. In isolation, both cercarial and pesticide exposure significantly decreased frog survival, development, and growth, and increased developmental malformations, such as scoliosis, kyphosis, and also edema and skin ulcers. The combination of cercariae and pesticides generally posed greater risk to frogs than either factor alone by decreasing survival or growth or increasing time to metamorphosis or malformations. The exception was that lone exposure to chlorpyrifos had higher mortality without than with cercariae. Consistent with mathematical models that suggest that stress should increase the impact of generalist parasites, the weight of the evidence from the field and laboratory suggests that ecologically relevant concentrations of agrochemicals generally increase the threat that trematodes pose to amphibians, highlighting the importance of elucidating interactions between anthropogenic activities and infectious disease in taxa of conservation concern.

The freshwater trematode Nanophyetus salmincola has been demonstrated to impair salmonid immune function and resistance to the marine pathogen Vibrio anguillarum, potentially resulting in ocean mortality. We examined whether infection by the parasite N. salmincola similarly increases mortality of juvenile Chinook Salmon Oncorhynchus tshawytscha when they are exposed to the freshwater pathogens Flavobacterium columnare or Aeromonas salmonicida, two bacteria that juvenile salmonids might encounter during their migration to the marine environment. We used a two-part experimental design where juvenile Chinook Salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., and then challenged with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare infection was higher in N. salmincola-parasitized fish than in nonparasitized fish. In contrast, cumulative percent mortality from A. salmonicida infection did not differ between N. salmincola-parasitized and nonparasitized groups. No mortalities were observed in the N. salmincola-parasitized-only and control groups from either challenge. Our study demonstrates that a relatively high mean intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae can alter the outcomes of bacterial infection in juvenile Chinook Salmon, which might have implications for disease in wild fish populations.

Tracking individual variation in the dynamics of parasite infections in wild populations is often complicated by lack of knowledge of the epidemiological history of hosts. Whereas the dynamics and development of Gyrodactylus salaris Malmberg, 1957, on Atlantic salmon, Salmo salar L., are known from laboratory studies, knowledge about infection development on individual wild fishes is currently sparse. In this study, the dynamics of an infection of G. salaris on individually marked Atlantic salmon parr was followed in a section of a natural stream. During the 6-week experiment, the prevalence increased from 3.3 to 60.0%, with an average increase in intensity of 4.1% day(-1) . Survival analyses showed an initially high probability (93.6%) of staying uninfected by G. salaris, decreasing significantly to 37% after 6 weeks. The results showed that even at subarctic water temperatures and with an initially low risk of infection, the parasite spread rapidly in the Atlantic salmon population, with the capacity to reach 100% prevalence within a short summer season. The study thus track individual infection trajectories of Atlantic salmon living under near-natural conditions, providing an integration of key population parameters from controlled experiments with the dynamics of the epizootic observed in free-living living populations., (© 2014 John Wiley & Sons Ltd.)

Fish-borne zoonotic trematodes (FZT) including heterophyids and opisthorchiids are prevalent in native and aquaculture fish in Southeast Asia. FZT are reported highly prevalent in juvenile aquaculture fish that belonged to the family Heterophyidae, particularly Haplorchis taichui. Aquaculture fish are reported to have varying levels of natural infection with H. taichui, but data for their susceptibility as well as resistance to infection are not available. The aim of this study was to determine the susceptibility of five aquaculture fish to H. taichui. Experimental infections were performed by exposing fish individually to 0 (control), 50, 100, or 200 H. taichui cercariae for 12 h. Metacercarial burden was measured at 45 days postinfection. Three out of five fish species, silver barb (Barbonymus gonionotus), common carp (Cyprinus carpio), and mrigal (Cirrhinus mrigala), were highly susceptible to H. taichui infection with percentage ranging from 93.33 to 100%. The Nile and red tilapia were not susceptible to infection. Among the susceptible fish species, silver barb had higher intensity of H. taichui metacercariae than common carp and mrigal (P < 0.001). Metacercarial burden significantly correlated with the dose of cercarial infection (P < 0.001). Our findings that common aquaculture fish species have varying degrees of susceptibility to H. taichui infection provide important information to reduce parasite transmission in aquaculture fish.

The importance of disease-mediated invasions and the role of parasite spillover as a substantial threat to the conservation of global biodiversity are now well known. Although competition between invasive sliders Trachemys scripta elegans and indigenous European turtles has been extensively studied, the impact of this invasive species on diseases affecting native populations is poorly known. During winter 2012-2013 an unusual event was detected in a population of Emys orbicularis (Linnaeus, 1758) inhabiting a pond system in Galicia (NW Spain). Most turtles were lethargic and some had lost mobility of limbs and tail. Necropsies were performed on 11 turtles that were found dead or dying at this site. Blood flukes belonging to the species Spirorchis elegans were found inhabiting the vascular system of 3 turtles, while numerous fluke eggs were trapped in the vascular system, brain, lung, heart, liver, kidney, spleen, and/or gastrointestinal tissues of all necropsied animals. Characteristic lesions included miliary egg granulomas, which were mostly found on serosal surfaces, particularly of the small intestine, as well as endocarditis, arteritis, and thrombosis. The most probable cause of death in the 3 turtle specimens which were also examined histologically was a necrotic enteritis with secondary bacterial infection associated with a massive egg embolism. The North American origin of S. elegans, the absence of prior recorded epizootics in the outbreak area, and the habitual presence of its type host, the highly invasive red-eared slider, in this area suggest a new case of parasite spillover resulting in a severe emerging disease.

Goldfish, Carassius auratus, widely distributed across Eurasia, is one of the earliest fish domesticated for ornamental purposes. A series of diseases break out due to high-density culture of goldfish, causing significant economic losses. Here, we report for the first time an infection of Gyrodactylus kobayashii associated with the high mortality of goldfish from a fish farm in Anziying township of Henan province, China, following their transfer to several aquariums. In the first 2 weeks after goldfish (n = 4200, weight 10.24 ± 1.45 g) transfer to aquariums, a total of 3335 goldfish were lost, representing 79.4 % of the initial stocked biomass. The examination showed that no other pathogens were found in this batch of fishes, except for G. kobayashii. The high burdens of G. kobayashii/fish (264.7, range 100-450) were the primary reason for the high mortality, although this was not the sole cause. In addition, the isolate of G. kobayashii from goldfish (C. auratus) in Henan province of central China was described. The morphological characterization was performed using morphometric measurements and drawings of opisthaptoral hard parts of the parasites. The molecular description was performed based on phylogenetic analysis of a reference DNA sequence spanning 5.8S and ITS-2. Importantly, the present study provides for the first time a full 25 point-to-point morphometric measurements and high-resolution images of attachment organ of G. kobayashii.

Atlantic salmon, Salmo salar L. parr (age 1+), infected by the monogenean ectoparasite Gyrodactylus salaris (Malmberg, 1957), were exposed to chlorine (Cl)-enriched water at three different concentrations: Cllow (0-5 μg Cl L(-1) ), Clmedium (18 μg Cl L(-1) ) and Clhigh (50 μg Cl L(-1) ). There was a negative correlation between G. salaris infections and the hypochlorite concentrations added. The parasite infection was eliminated by day 6-8 and day 2-4 in the groups Clmedium and Clhigh , respectively, while inhibition of G. salaris population growth was observed in the Cllow group. An important note to this matter, however, is that the G. salaris specimens observed at day 6 in Clmedium and at day 2 in Clhigh were all considered dead by subjective judgement. No mortality in the salmon parr was observed during the first 8 days of the experiment, demonstrating that Cl has a stronger effect on G. salaris than on the salmonid host. The differences in sensitivity between the parasite and the Atlantic salmon indicate that hypochlorite has a potential use as a parasiticide with a therapeutic margin. The low-dose sensitivity may imply that Cl pollution in urban areas may pose a greater risk towards biodiversity than previously assumed., (© 2014 John Wiley & Sons Ltd.)

Amphibians stand at the forefront of the global biodiversity crisis. The causes of their decline are diverse and include a rise in amphibian malformations due to various factors, especially trematode infection. However, linking amphibian mortality and morbidity with trematode infection has proven to be challenging due to the complex life cycle of the trematodes and the fact that trematodes are nonfastidious in their choice of definitive hosts. In Israel, the decline in local amphibian populations has been mostly attributed to the loss and degradation of wetlands and riparian habitats. Recently, however, there have been several reports of morbidity and mortality of tadpoles with signs of edema and malformations from various localities in Israel. We collected dead and morbid tadpoles and metamorphs of Hyla savignyi and Pelophylax bedriagae, and we showed that the morbidity and the deformations observed in the field are the result of infection by trematodes. We also isolated an echinostomatid trematode from the malformed and edematous tadpoles and from the freshwater snail Bulinus truncatus, all from the same site. We further succeeded in experimentally infecting H. savignyi tadpoles by echinostomatid cercariae that were shed from the snails, and we showed that infection had significantly increased the mortality rates of these tadpoles. The combination of high trematode prevalence and their pathogenic effects suggests that in nature, the effect of echinostome infection on amphibians may be substantial and could become an emerging disease in Israel.

Monogenean infections of commercially farmed fishes are responsible for significant economic losses. Garlic (Allium sativum) is a well-known spice which also possesses anti-microbial and anti-parasitical properties. The current work aimed to test the efficacy of garlic-based treatments against infection with monogenean sp. in the guppy (Poecilia reticulata). Clipped sections of tail fins of guppies heavily infected with Gyrodactylus turnbulli were exposed to aqueous garlic extract (7.5 to 30 mL L(-1)) and visually observed under a dissecting microscope. Results revealed that exposure to garlic caused detachment of parasite and cessation of movement indicating death. A positive correlation was seen between garlic concentration and time to detachment and death of parasites, which, at the highest concentration of 30 mL L(-1), occurred at 4.1 and 8.6 min, respectively. Bathing in aqueous garlic extract (7.5 and 12.5 mL L(-1)) was tested in guppies infected with G. turnbulli. Prior acute toxicity tests revealed the maximum tolerance levels of guppies to garlic extract to be 12.5 mL L(-1) for 1h. Bathing of infected fish in garlic extract (7.5 and 12.5 mL L(-1)) significantly (p<0.05) reduced infection prevalence and intensity as compared to the control. Oral treatments using dry garlic powder-supplemented diet were tested on guppies infected with G. turnbulli and Dactylogyrus sp. Fish were fed with food containing 10% and 20% dry garlic powder for 14 days. Groups fed with garlic supplemented diets showed significantly reduced (p<0.05) mean prevalence and mean intensity of parasites as compared to the control. Dietary application of garlic did not appear to affect palatability. Fresh crushed garlic was added at a level of 1 gL(-1) and applied as an indefinite bath for 14 days. This treatment was seen to significantly reduce (p<0.05) parasite prevalence and mean intensity as compared to the control. Histopathology revealed elevated muscular dystrophy in the 20% garlic-fed group, as compared to control. These findings demonstrate the potential of garlic as a natural alternative to currently used chemical treatments for monogenean sp. infection in the guppy., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Bithynia tentaculata is an aquatic invasive snail first detected in the upper Mississippi River (UMR) in 2002. The snail harbors a number of parasitic trematode species, including Sphaeridiotrema pseudoglobulus, that have been implicated in waterfowl mortality in the region. We assessed the capacity of S. pseudoglobulus cercariae to infect B. tentaculata and native snails found in the UMR. Four snail species (one invasive and three native) were individually exposed to S. pseudoglobulus larvae and all were successfully infected. A subsequent experiment examining infection patterns in invasive and native hosts exposed singly or in mixed treatments revealed no difference in parasite establishment among snail species. Our results add to our understanding of S. pseudoglobulus transmission and provide insight into processes underlying waterfowl disease in the UMR.

This study reports a disease outbreak among juvenile cobia (Rachycentron canadum) farmed in cages in the state of Rio de Janeiro, Brazil, caused by the dinoflagellate Amyloodinium ocellatum and the monogenean Neobenedenia melleni. Two thousand five hundred fish were stocked at 0.4 kg/m3 in a set of 12 m3 tanks, in autumn (mean weight 15.0 ± 7.3 g) and in winter (mean weight 43.0 ± 5.6 g). Freshwater baths were administered as a routine treatment, as the symptoms were detected followed by two collection samples. Firstly in May 2011 (n = 5) and secondly in September 2011 (n = 10). In the first sample, the prevalence of N. melleni on the body surface was 100% and the mean intensity was 42.0 ± 1.7, while in the second sample the prevalence was 60% with a mean intensity 3.0 ± 0.2 and mean abundance 1.8 ± 0.4. Amyloodinium ocellatum was only found in the second sample, at a prevalence 100% and mean intensity 46.8 ± 3.4. The cause of fish mortality was possibly associated with a decrease in fish resistance after the first contact with monogenean parasites, allied with respiratory difficulty caused by the presence of A. ocellatum in the gills.

Malformations and increased mortality due to infection by the digenetic trematode, Riberioa ondatrae have been reported for many species of amphibians. Severe malformations have also been reported in the Common Hourglass Tree Frog, Polypedates cruciger induced by pleurolophocercous cercariae in Sri Lanka in addition to the changes in the behaviour, development and survival of the host. We exposed pre-limb bud stage tadpoles (Gosner stages 25-26) of the Asian Common Toad, Duttaphrynus melanostictus to the same pleurolophocercous type cercariae under laboratory conditions. Molecular and morphological identification showed that these cercariae belonged Acanthostomum burminis infecting freshwater snakes as definitive hosts. These cercariae induced malformations (27.8%) and reduced survival to metamorphosis (53.8%). The magnitude of the effects increased with the dose of cercariae. Types of malformations were mainly axial, such as scoliosis and kyphosis. Severe limb malformations such as extra or missing limbs as reported for amphibians exposed to R. ondatrae were not observed in the D. melanostictus. Same authors reported a higher percentage of malformations previously when P. cruciger was exposed to the cercariae A. burminis compared to D. melanostictus. However, tadpoles of D. melanostictus, which are smaller compared to those of P. cruciger, experienced higher mortality than P. cruciger tadpoles. Trematode induced malformations and mortality in amphibians are highly variable and depend on multiple factors such as host species differences such as resistance to infection and tolerance, life-history characteristics such as size at metamorphosis and length of the metamorphosis period, and other factors such as size of the amphibian at the time of trematode exposure., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Body condition and parasite abundance were examined in two size classes of European bitterling Rhodeus amarus during the first overwintering period in two seasons (2007-2008 and 2009-2010). Body condition of large fish did not change during winter, and increased significantly in March. From November to February, small fish showed a decreasing trend in condition. Despite a significant increase in March condition of small fish only reached the same level as before winter. Total parasite abundance increased significantly in winter in both fish size classes, reflecting a seasonal increase in monogenean infection. Large fish were parasitized significantly more than small fish during winter, but only in small fish was a negative correlation between parasite infection and condition found and a significant decrease in parasite abundance recorded after wintering, indicating mortality of heavily infected individuals with low condition during the winter. A trend for higher overwinter mortality in small fish was found under semi-experimental conditions. The decrease in condition during the winter period in small fish may reflect faster energy depletion generally expected in smaller individuals. The results indicate that parasite infection may contribute to the overwinter mortality of 0+ year R. amarus, with a stronger effect in smaller individuals., (© 2013 The Authors. Journal of Fish Biology © 2013 The Fisheries Society of the British Isles.)

Background: Food-borne trematodiases are a group of neglected tropical diseases caused by liver, lung, and intestinal parasitic fluke infections. As part of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD 2010 study) and a WHO initiative, we assessed the global burden of human food-borne trematodiasis, as expressed in disability-adjusted life years (DALYs) for the year 2005., Methods: We systematically searched electronic databases for reports about human food-borne trematodiasis without language restriction, between Jan 1, 1980, and Dec 31, 2008. We used a broad search strategy with a combination of search terms and parasite and disease names. The initial search results were then screened on the basis of title, abstract, and, finally, full text. Relevant quantitative and qualitative data on human prevalence, morbidity, and mortality of food-borne trematodiasis were extracted. On the basis of available information on pathological and clinical appearance, we developed simplified disease models and did meta-analyses on the proportions and odds ratios of specified sequelae and estimated the global burden of human food-borne trematodiasis., Findings: We screened 33,921 articles and identified 181 eligible studies containing quantitative information for inclusion in the meta-analyses. About 56·2 million people were infected with food-borne trematodes in 2005: 7·9 million had severe sequelae and 7158 died, most from cholangiocarcinoma and cerebral infection. Taken together, we estimate that the global burden of food-borne trematodiasis was 665,352 DALYs (lower estimate 479,496 DALYs; upper estimate 859,051 DALYs). Furthermore, knowledge gaps in crucial epidemiological disease parameters and methodological features for estimating the global burden of parasitic diseases that are characterised by highly focal spatial occurrence and scarce and patchy information were highlighted., Interpretation: Despite making conservative estimates, we found that food-borne trematodiases are an important cluster of neglected diseases., Funding: Swiss National Science Foundation; Institute for Health Metrics and Evaluation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The association of Gyrodactylus anguillae Ergens, 1960 with the glass eel stage of Anguilla anguilla (L.) (total body length 61.4 ± 4.9 mm; range 55-70) is reported from the north-western Mediterranean coast of Spain for the first time. A sample of 12,600 glass eels, caught by professional fishermen operating in the mouth of the rivers Fluvià, La Muga and Ter (north-east Spain), was subject to mortalities of ∼ 1.75% of stock/day following transfer to a research facility. Subsequent losses over a 31-day period amounted to 56% of the initial stocked biomass. Although the moderate burdens of G. anguillae/host (20.2 ± 6; range 11-32) were the primary reason for a subsequent treatment, a simultaneous infection with Trichodina jadranica Raabe, 1958, Trichodina anguillae Wu, 1961 and Ichthyophthirius multifiliis Fouquet, 1876, makes it impossible to attribute the high mortality of glass eels in this case to a single pathogen. A histopathological examination of the gills of moribund fish showed them to be swollen, hyperplastic and necrotic. This study also redescribes G. anguillae, providing for the first time a full 27 character morphometric description of the attachment hooks, and importantly, a photographic record of the armature of the haptor and the male copulatory organ., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Populations of North American waterbirds, particularly lesser scaup, have been declining due to habitat disturbance, changing food resources, contaminants, bad water quality, and competition. However, epizootic diseases, including parasitism, may also play an important role in further decline. Trematode-associated mortality of migrating waterbirds, mainly American coot and lesser scaup, has been occurring in the Upper Mississippi River National Wildlife and Fish Refuge since 2002. We examined the levels of infective stages of Cyathocotyle bushiensis and Sphaeridiotrema globulus in the invasive, intermediate host snail, Bithynia tentaculata, during the fall of 2005 and compared these to infection levels in moribund or dead bird hosts. Our results show different infection levels of these 2 parasites in the 2 bird species; C. bushiensis is found more frequently in coot, and S. globulus is more common in scaup. This result is interesting because both bird species are presumed to forage on the same snail population and thus should be experiencing the same extent of exposure. These differences in infections could be attributed to differences in resources of gastrointestinal tracts of coot and scaup, or host resistance. Alternatively, differences in feeding behaviors of coot and scaup may also contribute to differential infections of the 2 trematodes.

Cardicola forsteri is a blood fluke that is highly prevalent among cultured southern bluefin tuna (SBT), Thunnus maccoyii, in South Australia. The role of C forsteri in annual SBT mortality outbreaks, which peak 6 to 12 weeks poststocking, is unknown. The objective of this study was to identify lesions unique to cultured SBT that died during a mortality event in 2009 and to determine the significance of C forsteri-associated lesions. Cultured SBT were sampled from 4 pontoons of a single site in Spencer Gulf that experienced a mortality epidemic that spanned 5 to 14 weeks poststocking. Study SBT comprised 7 that died during peak mortality, 27 that did not die, and 10 wild-caught (noncultured) SBT. All cultured SBT had branchitis and myocarditis due to C forsteri, whereas no life stages of C forsteri were histologically identified in any wild-caught SBT. Mortality was associated with the presence of severe branchitis (P<.005), and the odds of severe branchitis were 90 times greater for SBT that died than for SBT that were live caught during peak mortality (95% confidence interval, 5 to 1,684). In SBT that had died, no lesions other than those associated with C forsteri were of sufficient severity or physiologic significance to account for death. Other lesions common among cultured SBT included systemic granulocytic perivascular infiltrate, granulocytic gastric infiltrate, hepatic lipidosis, visceral granulomas, and branchial parasitic infestation. This study shows for the first time that a substantial proportion of poststocking mortality in cultured SBT is strongly associated with severe branchitis caused by C forsteri., (© The Authors 2011)

Centrocestusformosanus is a zoonotic trematode from Asia and has been mainly associated as cause of death of cultured fish. To identify pathogen trematode species in tilapia fry (Oreochromis niloticus) and to determine mollusks hosting these parasites, freshwater mollusks were collected from tilapia cultured ponds and experimental infections were carried out with tilapia fries and different mollusk species. A total of 907 freshwater mollusks were obtained from tilapia ponds and were identified to species level, four gastropods and one bivalve were determined: Melania tuberculata, Melanoides turricula, Pomacea flagellata, Haitia cubensis and Anodontiles luteola. For the first time, the presence of M. turricula and H. cubensis are reported in Costa Rica. Seven morphotypes of cercariae (Xifiodiocercaria, Equinostoma, Oftalmocercaria, Parapleurolofocercus, Cistocerca, Furcocercaria and Leptocercaria) parasitizing all five species of mollusks were found, all of distome type. Experimental exposure of tilapia fry to M. tuberculata demonstrated that the parapleurolofocercus morphotype found in the mollusk is in accordance with the finding of C. formosanus in tilapia fry. An abundance and mean intensity of 1018-1027 digeneans per gill in each exposed fish was determined. Centrocestus formosanus is reported for the first time in Costa Rica, for which the primary and secondary intermediate hosts were also determined.

Two different batches of meagre Argyrosomus regius were stocked in the same floating cage located in north-eastern Sardinia (western Mediterranean Sea). After 4 mo, the fish started showing non-specific disease signs, such as lethargy, emaciation, gill anaemia and mortality. In total 65 specimens of meagre of both stocks were sampled for parasitological and microbiological analysis, and only 1 species of parasite, the microcotylid monogenean Sciaenacotyle panceri, was found on the gills of caged meagre. No protozoan or other metazoan parasites were found and no bacteria or viruses were isolated. After the outbreak, a total mortality rate of 5 to 10% was estimated for the older fish stock, while no noteworthy mortality was recorded in younger fish (<2%). This is the first account of S. panceri on the gills of cultured meagre as causative agent of disease and mortality. The effects of the presence of this parasite emphasise the need for disease control strategies of cultured meagre, and investigating the transfer of microcotylids from wild to caged fish.

Seasonal dynamics of 2 trematode species, Cyathocotyle bushiensis and Sphaeridiotrema globulus, were assessed in relation to life history traits of the parasites and their hosts, as well as abundance of host species and abundance of infective stages. Both of these trematodes are associated with recurrent mortality of migrating waterbirds on the Upper Mississippi River National Wildlife and Fish Refuge. An invasive snail species, Bithynia tentaculata, serves as intermediate host for both trematode species. In total, 2,970 snails were collected at 2 study sites. Prevalence and mean abundance of the 2 trematode species varied among dates and was attributed to several factors, including migration patterns of definitive hosts, snail population dynamics, and seasonal changes in temperature. The surge of new infections of both parasites seems to be due to avian hosts foraging at this site during spring migration. The high prevalence and abundance of metacercariae among the snail population promote mortality among molluscivorous birds by increasing the probability of ingestion of a lethal dose. Additionally, mortality of non-molluscivorous birds can be explained by accidental ingestion of a couple of highly infected snails resulting in a lethal dose.

1. Within a community, different host species often exhibit broad variation in sensitivity to infection and disease. Because such differences can influence the strength and outcome of community interactions, it is essential to understand differential disease patterns and identify the mechanisms responsible. 2. In North American wetlands, amphibian species often exhibit extraordinary differences in the frequency of limb malformations induced by the digenetic trematode, Ribeiroia ondatrae. By coupling field studies with parasite exposure experiments, we evaluated whether such patterns were due to differences in (i) parasite encounter rate, (ii) infection establishment, or (iii) parasite persistence within hosts. 3. Field results underscored the broad variation in malformations and infection between host species; while nearly 60% (n = 618) of emerging American toads exhibited severe limb deformities such as bony triangles, skin webbings and missing limbs, fewer than 4% (n = 251) of Eastern gray treefrogs from the same pond were abnormal. Despite similarities in the phenology and larval development period of these species, they differed sharply in Ribeiroia infection. On average, toads supported 75x more metacercariae than did metamorphic treefrogs. 4. Experimental exposures of larval toads and treefrogs to a realistic range of Ribeiroia cercariae revealed strong differences in the sensitivity of these species to infection; exposed toads suffered elevated mortality (up to 95%), delayed metamorphosis, and severe limb malformations consistent with field observations. Treefrogs, in contrast, exhibited limited mortality and no malformations, regardless of exposure level. Ribeiroia cercariae were substantially less successful in locating and infecting Hyla versicolor larvae. 5. Our results indicate that the observed differences in infection and malformations owe to a lower ability of Ribeiroia cercariae to both find and establish within larval treefrogs, possibly stemming from a heightened immune response to infection. Because Ribeiroia is a highly pathogenic parasite with negative effects on larval and metamorphic amphibian survival, variation in infection resistance among species could have important implications for understanding patterns of species co-occurrence, competition, and community diversity.

Gyrodactylus pisculentus sp. n. is described from the head, body and fins of the northern pipefish, Syngnathus fuscus Storer (Syngnathidae) from the environs of Woods Hole, Massachusetts, USA. The new species is compared with Gyrodactylus shorti and Gyrodactylus syngnathi, species previously recorded from pipefish in the United States and Norway, respectively. Gyrodactylus pisculentus was frequently associated with mortality of northern pipefish held in quarantine at the Woods Hole Science Aquarium during 2002-2005. The clinical account of the infections and treatment protocols are presented.

In hybridogenetic systems, hybrid individuals are fully heterozygous because one of the parental genomes is discarded from the germinal line before meiosis. Such systems offer the opportunity to investigate the influence of heterozygosity on susceptibility to parasites. We studied the intensity of lung parasites (the roundworm Rhabdias bufomis and the fluke Haplometra cylindracea) in 3 populations of water frogs of the Rana lessonae-esculenta complex in eastern France. In these mixed populations, hybrid frogs (R. esculenta) outnumbered parental ones (R. lessonae). Despite variation in parasite intensity and demographic variability among populations, the relationship between host age and intensity of parasitism suggests a higher susceptibility in parentals than in hybrids. Mortality is probably enhanced by lung parasites in parental frogs. On the other hand, while parental frogs harboured higher numbers of H. cylindracea than hybrid frogs, the latter had higher numbers of R. bufonis. Despite such discrepancies, these results support the hybrid resistance hypothesis, although other factors, such as differences in body size, age-related immunity, differential exposure risks and hemiclonal selection, could also contribute to the observed patterns of infection.

Neodiplostomum seoulense is highly pathogenic and lethal to experimental mice; most worms are expelled within 2 mo of acquisition. In this study, T-helper (Th) cell immune responses were studied in N. seoulense-infected BALB/c mice. Spleen and mesenteric lymph node (MLN) cells of infected mice proliferated in response to parasite antigens; CD4+ T cells proliferated more than CD8+ T cells. Antigen-induced interferon (IFN)-gamma (a Th1 cytokine) secretion began to increase at day 7 postinfection (PI) in spleen and MLN cells, and this was maintained at day 28 PI, whereas interleukin (IL)-4 (a Th2 cytokine) secretion was somewhat lower. Similar results were observed for mRNA signals of IFN-gamma and IL-4. Antigen-specific serum total immunoglobulin (Ig)G, IgG1, IgM, and IgA levels (Th2-induced) were elevated from days 7 to 14 to day 28 PI, and IgG2a (Th1-induced) was elevated at days 21 to 28 PI. Interestingly, the numbers of macrophages (Th1- or Th2-induced), which were found to kill N. seoulense worms in vitro, increased remarkably during days 14-28 PI in spleens and small intestines of infected mice. This study shows that mixed Th1 and Th2 responses occur during the course of N. seoulense infection in BALB/c mice. Heavy infiltrations of macrophages in the small intestine may participate in host damage and worm expulsion.

The effect of echinostome infections on the survival of Rana pipiens tadpoles was examined in relation to developmental stage of tadpoles. Individual tadpoles of Gosner stages 25, 27, 32-33, and 37-39 were exposed to 1 of 4 levels of cercariae (0, 20, 50, or 100). Only tadpoles at stage 25, the earliest stage infected, died within a 5-day experimental period. This stage-specific mortality rate could be explained, in part, by the stage-specific location of encystment of cercariae, which was documented in a separate experiment. In accordance with kidney development, cercariae predominately encysted in the pronephroi during early stages of tadpole development (stages 25 through 31-32) and only in the mesonephroi and associated ducts at later stages (stages 37 through 46). As the mesonephros develops, renal capacity presumably increases. Thus, tadpoles died only when metacercariae concentrated in the functional portion of the kidney with the most limited renal capacity. As tadpoles aged, they also became less susceptible to infections. On average, 69.5% of cercariae that were exposed to stage 25-26 tadpoles successfully encysted. compared with only 8.4% of cercariae exposed to stage 37-38 tadpoles. Exposures of metamorphic frogs (poststage 46) to cercariae revealed that these individuals can become infected with echinostomes. Collectively, our data highlight the host stage-dependent dynamics of tadpole-echinostome interactions.

Annual, seasonal, and interlake variation in prevalence and intensity of Ornithodiplostomum ptychocheilus (Faust) metacercariae was assessed in populations of fathead minnows (Pimephales promelas) collected from 4 lakes in north-central Alberta. Mean metacercariae intensity in young-of-the-year minnows varied extensively (5-123 metacercariae/host) among year, month, and lakes. In 2 of the lakes, prevalence always reached 100%, and mean intensity always peaked in September or October. The high spatial and annual variation in metacercarial recruitment was partly attributable to variation in host size. but variation in water depth, temperature, snail densities, and bird visitation likely also played a role. A laboratory experiment demonstrated that host and metacercariae survival was intensity-independent during a period of simulated winter. Thus, metacercariae recruited in the fall survive until the following spring.

The genetic influence on host susceptibility to Neodiplostomum seoulense infection and fatality of the host was studied in 3 inbred strains of mice (BALB/c [H-2d], C3H/He [H-2k], and C57BL/6 [H-2b]). The survival of the mice, worm expulsion kinetics, worm size, number of eggs produced per day (EPD), and number of uterine eggs were observed from day 1 to day 40 postinfection (PI) with 100 or 200 metacercariae per mouse. Infection with N. seoulense was highly lethal to all 3 strains of mice, but the lethality was dose-dependent and varied according to the genetic backgrounds of the mice. The C3H/He mice exhibited the highest mortality, the lowest worm burdens and EPD, and the quickest expulsion of worms. It is suggested that different genetic backgrounds of mice appear to affect the host's capacity to expel N. seoulense and the fatality of the hosts themselves.

Monthly data collected during 1993 in the Vaccarès lagoon (Rhône Delta, France) were used to investigate the influence of the trematode Aphalloïdes coelomicola on the survival of the common goby Pomatoschistus microps, which acts both as second intermediate and definitive host. Prevalence of the parasite did not increase with fish size in either sex. Mean abundance and variance to mean abundance ratio gave evidence that the trematode could have an impact on host mortality. Gonad weight of parasitized females was reduced and the parasite seemed to provoke a gonadal regression above a certain parasitic biomass. Fecundity and egg diameters did not differ between parasitized and unparasitized females, we thus hypothesize that the energy content of eggs (composition in lipids and proteins) was probably modified as an adaptation to energy losses. Prevalence of infection in males compared to females and influence of the parasite on host survival and reproductive effort are discussed.

Although the effect exerted on fish by fluke cercariae developing in fish as intermediate hosts is fairly well known, cercariae of flukes requiring intermediate hosts other than fish occur in fish ponds in much higher numbers. To study the effect of these latter cercariae on fish fry, a few days old common carp fry (Cyprinus carpio) were exposed to cohorts of xiphidiocercariae or echinocercariae shed by one or two snails acting as intermediate hosts. The observed pathogenicity of these cercariae was compared to the well-known pathogenic effect of Diplostomum cercariae. Although Diplostomum cercariae proved to be the most pathogenic, occasionally also the other two types of cercariae could kill the common carp fry by invading their body. Xiphidiocercariae ingested by the fish penetrated the intestinal wall and developed into metacercariae within the body of the fry. They could also get into the musculature through the skin. The much larger echinocercariae invaded the gill cavity where they became encapsulated, thus hampering gill function and leading to fish mortality due to oxygen deficiency. Diplostomum cercariae were more pathogenic than the cercariae of the other two flukes in that they were shed by the intermediate host in very large numbers. Although much less xiphidiocercariae and echinocercariae emerged from the intermediate host snails than Diplostomum cercariae, the former could kill the fish fry already in lower numbers. These findings prove that fluke cercariae nonspecific to fish may exert pathogenic effects on fish, and presumably contribute to the early mortality of young fry placed out into fish ponds.

Pathogenicity and lethality of Neodiplostomum seoulense to various strains of mice (mast cell-deficient W/Wv, their normal littermate +/+, C57BL/6, BALB/cA, C3H/HeJ), and a hybrid (BALB/cA x C3H/HeJ)F1 were investigated. When the mice were infected orally each with 200 metacercariae, their abdomen became distended, and all mice died by day 23 postinfection (PI) except BALB/cA, which were severely weakened but recovered after 28 days. Even a smaller infection dose of 25 metacercariae was highly lethal to C3H/HeJ mice. Despite treatment with praziquantel (3 mg/mouse) on day 10 PI, 80% of C57BL/ 6 mice did not recover and died. After day 14 PI, the whole intestine of C57BL/6 mice was contracted and significantly shortened in length, and charcoal meal transit was significantly faster compared with uninfected controls. After incubation in papaverine, the contracted intestines of C3H/HeJ mice did not relax, suggesting that the change is irreversible. In conclusion, N. seoulense has the potential to kill most strains of mice by causing irreversible damage to their intestine. The effects of mucosal mast cells on the survival rate of mice were negligible, because both W/Wv and +/+ mice died around day 20 PI.

Four outbreaks of gas bubble disease were encountered among farmed fish in Saudi Arabia. Two of them occurred among subadult (52.5 g) saltwater tilapia (Oreochromis spilurus), the first affecting about 50 per cent of the stock and resulting in about 30 per cent mortality, and the second affecting about 25 per cent of the population with about 5 per cent mortality. Another outbreak occurred among adult (270 g) brackish water (0.5 per cent salinity) tilapia (Oreochromis niloticus), affecting about 40 per cent of the population with about 25 per cent mortality. The fourth outbreak occurred among three-month-old (15 g) grouper (Epinephelus fuscogutiatus) and resulted in 10 per cent mortality. In all cases the total water gas pressure ranged between 111.2 and 113.4 per cent saturation and nitrogen was supersaturated while oxygen was undersaturated. The outbreaks were alleviated by reducing the gas pressure by splashing the source water or by switching to a source of water with lower gas pressure. However, in O niloticus the conditions of gas supersaturation resulted in a heavy infection by monogenetic trematodes which was treated with formalin at 40 mg/litre for seven hours on five successive days.

In five experiments, common carp (Cyprinus carpio) fry with Dactylogyrus vastator (Monogenea) infection of different severity, as well as parasite-free fry, were placed into aquaria containing malachite green solution of 0.5-10 mg/l concentration. Most of the fish died within 6 hours. The correlation between the time of death and helminth infection was studied. The experiments demonstrated that in malachite green solution the carp with the heaviest helminth infection died first while the less severely infected specimens survived for the longest time.

To identify causes of mortality in young captive crocodiles, detailed necropsy and laboratory examination was done on 54 (30 Crocodylus porosus, 22 C. novaeguineae, 2 of unrecorded species). Although multiple infections often confounded interpretation it was concluded that the major infectious diseases, of approximately equal importance, were coccidiosis, bacterial septicaemia with Gram-negative organisms, and metazoan parasitism including ascariasis and pentastomiasis. A range of other lesions and agents was recognised, including keratitis, enteritis of unknown aetiology, non-suppurative encephalitis, traumatic peritonitis and trematodes located in renal tubules, gut and blood vessels. Some crocodiles in poor condition had only mild lesions associated with metazoan parasites and the cause of death or illness could not be clearly determined, although it was considered likely that adaptation failure was a contributing factor.

Over the past 20 yr, recurrent late-summer mortality of dabbling ducks in southern Québec has been attributed to the digenean parasite Cyathocotyle bushiensis. This study attempted to determine whether this parasite was still implicated in the epizootics, and examined aspects of the ecology of the parasite in the definitive host. Comparison of prevalence and mean intensity of infection of all species of Digenea between salvaged carcasses and hunter-shot ducks revealed that C. bushiensis and a second digenean Sphaeridiotrema globulus were both significantly more prevalent in salvaged ducks. Mean intensity of infection was consistently higher for S. globulus in the salvaged carcasses than in hunter-shot ducks. These data provide strong circumstantial support for the hypothesis that C. bushiensis continues to contribute to the annual mortality, but also strongly suggest that S. globulus may be involved. Comparison of the relative numbers of each species of duck between the salvaged carcass sample and the hunter-shot sample revealed that blue-winged teal (Anas discors) occurred significantly more frequently in the salvaged carcass sample whereas wood ducks (Aix sponsa) were common in the hunter-shot sample but were never found in the salvaged carcass sample. A comparison of susceptibility to experimental infection with C. bushiensis revealed that mallards (Anas platyrhynchos), black ducks (Anas rubripes), blue-winged teal, pintail (Anas acuta) and lesser scaup (Aythya affinis) had similar susceptibilities but that wood ducks were significantly less susceptible to infection. Experimental infections also showed that age of mallards significantly influenced the number of parasites obtained 7 days postinfection; ducklings were most heavily infected. Significant differences were detected in the gross pathology caused by C. bushiensis among duck species. Data obtained from sentinel ducks placed in the field for 24 hr revealed that transmission of both C. bushiensis and S. globulus occurs at least between mid-May and mid-September and that large, potentially lethal, infections can be acquired in as little as 24 hr. Finally evidence is presented to suggest that ducks acquire resistance to challenge infections of C. bushiensis.

Ulcerative hemorrhagic enteritis of swans induced by Sphaeridiotrema globulus is enzootic at Lake Musconetcong in northern New Jersey. It accounted for a calculated 250 deaths of wild mute swans (Cygnus olor) between 1970 and 1980. The swans died primarily in the winter and early spring. Sites of infection included the jejunum and ileum. Trematodes singly or in groups ulcerated the intestines. Severe hemorrhage from damaged submucosal capillaries provided a blood meal for the parasites and probably precipitated the anemia observed. The intensity of fatal infections ranged from 20 to 870 parasites. A mononuclear periportal hepatitis and secondary nodules in the spleen were commonly observed. Affected swans exhibited muscular weakness and appeared to have died from hypovolemic shock or some complication associated with it.

Mortality of pelyad (Coregonus peled) caused by Tetraonchus alaskensis took place in winter 1973 in the Voikara and Syn rivers (the Ural tributaries of the Lower Ob) during anadromous and catadromous migrations. The highest infection rate was observed in older fished. Up to 2330 specimens of the parasite were recorded from the gills of one pelyad. A low infection rate with T. alaskensis was recorded from Coregonus nasus, C. lavaretus pidgian and Stenodu. The mortality coincided with the period of the greatest abundance of the Ob pelyard stock.