Major drivers of biodiversity loss and their impacts on helminth parasite populations and communities.

The world's biodiversity is in peril. The major threats to biodiversity globally are habitat change, overexploitation, climate change, invasive species and pollution. Not only do these stressors impact free-living organisms, but they affect parasitic ones as well. Herein, this perspective examines the potential consequences of these anthropogenic perturbations on helminth populations and communities, with emphasis on significant developments over the past decade. Furthermore, several case studies are examined in more detail for each of these threats to biodiversity. While effects are widespread and diverse, for the most part all these environmental stressors have negative effects on parasite populations and communities. Those parasites with complex life cycles that are trophically transmitted are often more at risk, although larval parasites with a wide host spectrum, and directly transmitted ectoparasites, appear less threatened and may even benefit. However, differential effects on hosts and parasites, on parasite life cycle stages and on host-parasite interactions made specific predictions difficult and context-dependent. Experimental laboratory and mesocosm studies on specific parasites that test effects on the different life cycle stages, hosts and host-parasite interactions, permitting the determination of net effects of an environmental stressor, yield insightful and sometimes counterintuitive results, although they remain a simplification of real-world complexity. Recent advances in the use of parasites as bioindicators of effects also are discussed.

Infection of Diplostomum spp. in invasive round gobies in the St Lawrence River, Canada.

The round goby (Neogobius melanostomus) is a successful invader of the Great Lakes-St Lawrence River basin that harbours a number of local parasites. The most common are metacercariae of the genus Diplostomum. Species of Diplostomum are morphologically difficult to distinguish but can be separated using molecular techniques. While a few species have been sequenced from invasive round gobies in this study system, their relative abundance has not been documented. The purpose of this study was to determine the species composition of Diplostomum spp. and their relative abundance in round gobies in the St Lawrence River by sequencing the barcode region of cytochrome c oxidase I. In 2007-2011, Diplostomum huronense (=Diplostomum sp. 1) was the most common, followed in order by Diplostomum indistinctum (=Diplostomum sp. 4) and Diplostomum indistinctum sensu Galazzo, Dayanandan, Marcogliese & McLaughlin (2002). In 2012, the most common species infecting the round goby in the St Lawrence River was D. huronense, followed by D. indistinctum and Diplostomum gavium (=Diplostomum sp. 3). The invasion of the round goby in the St Lawrence River was followed by a decline of Diplostomum spp. in native fishes to low levels, leading to the previously published hypothesis that the presence of the round goby has led to a dilution effect. Herein, it is suggested that despite the low infection levels in the round goby, infections still may lead to spillback, helping to maintain Diplostomum spp. in native fishes, albeit at low levels.

Distribution of parasites of slimy sculpin Cottus cognatus Richardson, 1836 (Scorpaeniformes: Cottidae) in the Athabasca drainage, Alberta, Canada, and their relation to water quality.

The composition and diversity of parasite communities are useful tools to characterise ecosystem health and integrity. Environmental disturbances may affect parasite infection in fish directly, by their effects on the free-living stages, or indirectly, on the intermediate hosts. Slimy sculpins, Cottus cognatus, a small fish inhabiting cold waters of North America, have been considered as sentinels due to their limited mobility, often occupying relatively small areas throughout their lives and thus reflecting the local environment. Ninety-six specimens of C. cognatus were sampled from four tributaries of the Athabasca River to assess patterns of helminth parasite community structure in this fish and to study the composition and diversity of its parasite communities in relation to water quality. The localities included single samples from High Hills, Horse and Dunkirk rivers, and two from the Steepbank River. Twelve metazoan parasite species were found, most of them being larval forms. Significant differences occurred in the structure and composition of parasite assemblages of sculpins from the tributaries, although similarities were observed in connected and nearby sites. Parasite communities were influenced mainly by a combination of local environmental conditions, distance and connectivity, and were separated based on the distribution and abundance of autogenic and allogenic parasites. Water quality appeared to influence the distribution of trematode species that use gastropods as intermediate hosts, while proximity and connectivity of sites led to sharing allogenic parasite species in slimy sculpin.

Parasite community similarity in Athabasca River trout-perch (Percopsis omiscomaycus) varies with local-scale land use and sediment hydrocarbons, but not distance or linear gradients.

Parasite communities have been shown to be structured by processes at scales ranging from continental to microhabitat, but few studies have simultaneously considered spatial and environmental variables, measured at different scales, to assess their relative influences on parasite abundance, species richness, and community similarity. Parasite abundance, diversity, and community similarity in Athabasca River trout-perch (Percopsis omiscomaycus) were examined in relation to water quality, substrate profile, metal and organic compound levels in water and sediment, and landscape use patterns at different scales, as well as distance among sites and upstream-downstream position along the river. Although species richness did not differ among sites, there were significant differences in abundance of individual taxa and community structure. We observed a shift from communities dominated by larval trematodes Diplostomum spp. to domination by gill monogeneans Urocleidus baldwini, followed by a reversion further downstream. Variations in the abundance of these taxa and of overall community similarity were strongly correlated with sediment hydrocarbons (alkanes and polycyclic aromatic hydrocarbons (PAHs)) as well as landscape use within 5 km of study sites. No correlations were noted with any other predictors, indicating that parasite populations and communities in this system were likely primarily influenced by habitat level and landscape-scale filters, rather than larger-scale processes such as distance decay or river continuum effects.

Completion of the life cycle of Tylodelphys mashonense (Sudarikov, 1971) (Digenea: Diplostomidae) with DNA barcodes and rDNA sequences.

The life cycle of Tylodelphys mashonense (Digenea: Diplostomidae), whose metacercariae occur in the cranial cavity of the widely cultivated catfish Clarias gariepinus, was resolved by the application of molecular markers. Both COI barcodes and ITS sequences obtained from diplostomid-like cercariae infecting Bulinus spp. from Mindu Dam, Morogoro, matched those acquired from metacercariae from the catfish C. gariepinus, and those from adult T. mashonense from the grey heron Ardea cinerea and the white egret Egretta alba. The success in linking the life cycle stages of T. mashonense using molecular tools highlights the usefulness of this approach in resolving the complex life cycles of digeneans in the absence of experimental establishment.

Morphometric and molecular analyses of Tylodelphys sp. metacercariae (Digenea: Diplostomidae) from the vitreous humour of four fish species from Lake Naivasha, Kenya.

Even in the relatively well-characterized faunas of the developed world, it is difficult to discriminate species of metacercariae in the Diplostomidae using morphology, infection site or host use. The taxonomy, diversity and ecology of diplostomids infecting freshwater fishes in the African continent are particularly poorly known, but recent morphometric and genetic studies have revealed four species of diplostomids in the eyes and brains of siluriform fishes. In the present study, diplostomid metacercariae were collected from the eyes of 288 fish comprising two species within the Cyprinidae (Cyprinus carpio, n = 145, and Barbus paludinosus, n = 67), two Cichlidae (Oreochromis leucostictus, n = 56, and Tilapia zillii, n = 18) and one Centrarchidae (Micropterus salmoides, n = 2) caught in Lake Naivasha, Kenya. Morphometric (14 characters and 8 indices in 111 specimens) and molecular (sequences from the barcode region of the cytochrome c oxidase 1 gene in 11 specimens) data were used to discriminate species. All fish species except B. paludinosus were infected with Tylodelphys metacercariae that were initially separated into two types differing mainly in body length. However, this morphological distinction received only intermediate support in quantitative morphological analysis and molecular data indicated that both morphotypes were conspecific. All the specimens therefore are inferred to belong to a single unidentified species of Tylodelphys, which is not conspecific with any other diplostomid for which comparable molecular data are available, including four diplostomid species known from siluriform fish in Nigeria and Tanzania.

Growth and condition of juvenile coho salmon Oncorhynchus kisutch relate positively to species richness of trophically transmitted parasites.

The aims of this study were first, to test the hypothesis that metrics of fish growth and condition relate positively to parasite species richness (S(R)) in a salmonid host; second, to identify whether S(R) differs as a function of host origin; third, to identify whether acquisition of parasites through marine v. freshwater trophic interactions was related to growth and condition of juvenile salmonids. To evaluate these questions, species diversity of trophically transmitted parasites in juvenile coho salmon Oncorhynchus kisutch collected off the coast of the Oregon and Washington states, U.S.A. in June 2002 and 2004 were analysed. Fish infected with three or more parasite species scored highest in metrics of growth and condition. Fish originating from the Columbia River basin had lower S(R) than those from the Oregon coast, Washington coast and Puget Sound, WA. Parasites obtained through freshwater or marine trophic interactions were equally important in the relationship between S(R) and ocean growth and condition of juvenile O. kisutch salmon.

Estimating diet in individual pumpkinseed sunfish Lepomis gibbosus using stomach contents, stable isotopes and parasites.

The diets of 99 pumpkinseed sunfish Lepomis gibbosus from a pair of small, adjacent lakes in Ontario, Canada, were estimated from their stomach contents, trophically transmitted parasites and stable isotopes of carbon and nitrogen in fish tissue. The three methods provided virtually unrelated information. There was no significant correlation in the importance of any prey item across all three methods. Fish with similar diets according to one method of estimating diet showed no tendency to be similar according to other methods. Although there was limited variation in fish size and the spatial scale of the study was small, both fish size and spatial origin showed comparatively strong associations with diet data obtained with all three methods. These results suggest that a multidisciplinary approach that accounts for fish size and spatial origins is necessary to accurately characterize diets of individual fish.

Trichodinid (Ciliophora: Trichodinidae) Infections in Perch (Perca fluviatilis) experimentally exposed to pulp and paper mill effluents.

Wild-caught European perch (Perca fluviatilis) were exposed in the laboratory to untreated bleached pulp and paper mill effluent in two separate experiments. The first experiment was conducted at 7-8°C using effluent concentrations of 5 and 10%, and the second experiment was conducted at ambient river temperature of 4-20°C using an effluent concentration of 1%. Trichodinid ciliates were identified and enumerated at the end of the exposure using a mucus subsampling technique from gill and skin as well as a formalin immersion technique, which provided total counts on each fish. Four different trichodinid species were identified on the fish. Prevalence of infection, mean number, and mean density of Trichodina spp. decreased on fish exposed to effluents compared with controls. Prevalence of infection, mean number, and mean density of Trichodinella epizootica decreased on fish exposed to 5% and 10% effluents but increased on fish exposed to 1% effluents compared with controls. These results demonstrate that trichodinid ciliates vary in their susceptibility to at least certain types of contaminants and cautions against using trichodinids as environmental indicators without delineating species.

Temporal dynamics of epidermal responses of guppies Poecilia reticulata to a sublethal range of waterborne zinc concentrations.

This study assessed the histological changes in the epidermis of guppies Poecilia reticulata induced by waterborne zinc (Zn). Laboratory-reared P. reticulata fry were maintained individually in separate vessels containing artificial water (8 microg l(-1) Zn) to which 0, 15, 30, 60 or 120 microg l(-1) Zn was added. Their epidermal response to Zn was monitored regularly over 4 weeks. Compared with controls, mucus was rapidly released and mucous cell numbers decreased at all concentrations. Thereafter mucous release, epidermal thickness, numbers and size of mucous cells fluctuated at a rate that varied with Zn concentration, but fluctuations declined after day 18. Results clearly highlight the dynamic nature of the epidermal response to sublethal concentrations of waterborne Zn. In general, low concentrations of Zn induced a rapid response with reduced numbers and size of mucous cells and shift in mucin composition, and a subsequent thickening of the epidermis. Epidermal thickness and mucous cell area fluctuated over time but were normal after a month of exposure to low Zn concentrations. The number of mucous cells, however, remained low. Virtually all mucous cells from fish maintained in 15 and 60 microg l(-1) Zn contained acidic mucins throughout the month, whereas fish maintained at 30 microg l(-1) Zn responded by production of neutral mucins during the first 12 days followed by a mixture of neutral and acidic mucins. At 120 microg l(-1) Zn, the most dramatic effects were the gradual but sustained decrease in numbers and area of mucous cells, and the shift to acidic mucins in these cells. Thus, as concentration of Zn increased, the epidermal responses indicated a disturbed host response (dramatic decline in mucous cell numbers, with mixed composition of mucins), which may have been less effective in preventing Zn uptake across the epithelium.

The impact of climate change on the parasites and infectious diseases of aquatic animals.

Climate change is predicted to have important effects on parasitism and disease in freshwater and marine ecosystems, with consequences for human health and socio-economics. The distribution of parasites and pathogens will be directly affected by global warming, but also indirectly, through effects on host range and abundance. To date, numerous disease outbreaks, especially in marine organisms, have been associated with climatic events such as the El Niño-Southern Oscillation. In general, transmission rates of parasites and pathogens are expected to increase with increasing temperature. Evidence suggests that the virulence of some pathogens and parasites may also increase with global warming. The effects of climate change on parasites and pathogens will be superimposed onto the effects of other anthropogenic stressors in ecosystems, such as contaminants, habitat loss and species introductions. This combination of stressors may work cumulatively or synergistically to exacerbate negative effects on host organisms and populations. Climatic effects on parasites and diseases of key species may cascade through food webs, with consequences for entire ecosystems.

Effects of cadmium exposure on embryogenesis of Stagnicola elodes (Mollusca, Gastropoda): potential consequences for parasite transmission.

Experiments on the toxicity of cadmium (Cd(2+)) to the embryonic development of Stagnicola elodes (Mollusca, Gastropoda), obligatory first intermediate host of numerous trematodes of pathogenic importance, were carried out as part of a study on the effects of metal pollution on host-parasite relationships. Freshly laid snail eggs were exposed to Cd concentrations of 0, 0.02, 0.2, and 2.0 mg Cd(2+)/L, and survival and embryogenesis were examined for 30 days. Mean survival time (+/- SD) of the control group was 23.1 (+/- 5.3) days compared with 10.1 (+/- 3.2) at 0.02 mg Cd(2+)/L, 3.9 (+/- 0.7) at 0.2 mg Cd(2+)/L, and 1.1 (+/- 0.08) at 2.0 mg Cd(2+)/L. Mortality patterns of all test groups differed significantly from each other, demonstrating that the percentage of surviving individuals at any given time was inversely related to Cd concentration. Concentration-dependent effects of Cd exposure on snail embryogenesis were noted. While embryos of the control group developed properly and started hatching on day 16, eggs exposed to 0.02 mg Cd(2+)/L exhibited a prolonged gastrula period and failed to hatch. Eggs in the 0.2 mg Cd(2+)/L group were blocked in the gastrula stage on day 5, whereas individuals exposed to 2.0 mg Cd(2+)/L died in the morula stage on the second day. Data showed that Cd severely affects S. elodes embryogenesis. By implication, Cd contamination at concentrations >or=0.02 mg Cd(2+)/L will thus decrease transmission success of various trematodes by decreasing intermediate host snail abundance.

Fecundity of sealworm (Pseudoterranova decipiens) infecting grey seals (Halichoerus grypus) in the Gulf of St. Lawrence, Canada: lack of density-dependent effects.

Uterine egg counts were determined by means of an electronic particle counter in individual female sealworm, Pseudoterranova decipiens (Nematoda: Ascaridoidea: Anisakidae) collected from grey seals (Halichoerus grypus) from Anticosti Island, northern Gulf of St. Lawrence, in August and September 1992. Mean number of eggs per female was 54,916.9 (+/- 51,866.4), with a maximum of 300,000. Egg counts were strongly correlated with worm length, and less so with host sex and age, sealworm sex ratio, and densities of Anisakis simplex and Phocascaris sp. No effect of sealworm density on worm egg counts was determined and competitive effects by Contracaecum osculatum on sealworm fecundity were not observed. Analyses demonstrated that variation in sealworm fecundity was greater among seal hosts within groups than between groups for host age and sex, density of sealworm, and density of C. osculatum. No evidence for density-dependent regulation of sealworm fecundity could be found. Variations in worm egg counts are attributed to individual host effects on worm growth and fecundity.

Effects of agricultural pesticides on the immune system of Xenopus laevis and Rana pipiens.

Over the last 30 years, there have been mass declines in diverse geographic locations among amphibian populations. Multiple causes have been suggested to explain this decline. Among these, environmental pollution is gaining attention. Indeed, some chemicals of environmental concern are known to alter the immune system. Given that amphibians are frequently exposed to agricultural pesticides, it is possible that these pollutants alter their immune system and render them more susceptible to different pathogens. In this study, we exposed two frog species, Xenopus laevis and Rana pipiens, for a short period of time to a mixture of pesticides (atrazine, metribuzine, endosulfan, lindane, aldicarb and dieldrin) representative in terms of composition and concentrations to what it is found in the environment of the southwest region of the province of Quebec. The pesticides were known to be present in surface water of many tributaries of the St. Lawrence River (Quebec, Canada). Our results demonstrate that the mixture of pesticides could alter the cellularity and phagocytic activity of X. laevis and the lymphocyte proliferation of R. pipiens. Taken together, these results indicate that agricultural pesticides can alter some aspects of the immune response in frogs and could contribute to their global decline by rendering them more susceptible to certain infections.

Effects of cadmium upon longevity of Diplostomum sp. (Trematoda: Diplostomidae) cercariae.

Laboratory experiments were conducted to examine the impact of cadmium (Cd) on survival of Diplostomum sp. cercariae. Freshly emerged cercariae were exposed to Cd solutions at concentrations ranging between 0.2 and 200 microg/l. The mortality patterns and mean survival times (MSTs) of the exposed parasites were determined by the Kaplan-Meier estimator and were compared (Log-Rank test) to corresponding patterns and estimators of the unexposed controls. Cd concentrations >20 microg/l caused statistically significant changes in the mortality pattern of Diplostomum sp. cercariae and reduced MST by at least five hours compared to the control group, where MST was 52 h. The results show that longevity of the free-living Diplostomum sp. cercariae can be reduced by direct exposure to heavy metals. Cd contamination of aquatic habitats thus potentially can have an impact on parasite populations and communities.

Occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata).

A study was conducted in 1999 to determine the occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata). Concentrations of selected contaminants were measured in surface water, municipal effluent, sediments and mussels. Analyses were performed on 4-tert-octylphenol (4-t-OP), 4-n-nonylphenol (4-n-NP), nonylphenol polyethoxylates (NP(1-16)EO), nonylphenol-mono and di-ethoxycarboxylic acids (NP(1)EC and NP(2)EC), and octylphenol-mono and di-ethoxycarboxylic acids (OP(1)EC and OP(2)EC). Mussels (Elliptio complanata) taken from a reference lake were placed in cages and submerged for 62 days at two sites in the St. Lawrence River, 1.5 km upstream and 5 km downstream of the outfall of a municipal wastewater treatment plant. The results showed that many of the target chemicals were present in all matrices studied: in water, at ppt and ppb levels, and reaching ppm levels in sediments and mussels. Concentrations of these contaminants were higher in matrices sampled at the downstream site than in those drawn at the site upstream of the Montreal effluent outfall, especially in sediments. Likewise, the slight, but not significant, bioconcentration of certain alkylphenol polyethoxylates (AP(n)EO) in the mussels was more noticeable at the downstream site than at the upstream site.

Larval nematodes infecting Amphiporeia virginiana (Amphipoda: Pontoporeioidea) on Sable Island, Nova Scotia.

Three species of larval nematodes were found infecting the nearshore subtidal amphipod Amphiporeia virginiana collected from beaches on Sable Island, Nova Scotia. Five and 2 sealworms, Pseudoterranova decipiens s.l., were found in 2,020 and 2,044 amphipods, respectively, collected in 1991 and 1992, dissected, and examined microscopically. Two sealworms, 1 Paracuaria adunca and 1 Ascarophis sp., were found in 5,157 amphipods collected in 1992 and enzymatically digested. Infection with each species constitutes a new host record. These 3 nematodes possess the capability to infect a wide variety of invertebrate intermediate hosts. The shoreline niche of A. virginiana places it in close proximity to the sealworm's definitive host, the grey seal, which hauls out on Sable Island in abundance. The amphipod's distribution also places it in the vicinity of both the definitive hosts of Ascarophis sp. (marine fish) and P. adunca (piscivorous birds).

Aeginina longicornis (Amphipoda: Caprellidea), new intermediate host for Echinorhynchus gadi (Acanthocephala: Echinorhynchidae).

Cystacanths of the acanthocephalan Echinorhynchus gadi were found infecting the caprellid amphipod Aeginina longicornis on Banquereau, an offshore bank on the Scotian Shelf east of Nova Scotia, Canada. Two of 229 amphipods (0.9%) were each infected with a single male cystacanth. This is the first report of an intermediate host for E. gadi in the western Atlantic Ocean and the first record of A. longicornis as an intermediate host for any helminth. The cystacanths are described.

Seasonal occurrence of Lernaea cyprinacea on fishes in Belews Lake, North Carolina.

Prevalence, abundance, and mean intensity of Lernaea cyprinacea were monitored on 3 species of fishes from June 1984 through August 1986 in Belews Lake, North Carolina. Copepods were observed parasitizing fishes only during summer, when water temperatures exceeded 25 C. Infection levels in mosquitofish (Gambusia affinis) and red shiners (Notropis lutrensis) were very low, but they were much higher in fathead minnows (Pimephales promelas). Infection levels do not appear to be related to the size of the host. Host differences in parasite prevalence may be due to the host habitats.

Myxozoan communities parasitizing Notropis hudsonius (Cyprinidae) at selected localities on the St. Lawrence River, Quebec: possible effects of urban effluents.

Late summer myxozoan infra- and component communities parasitizing 73 Notropis hudsonius at 5 sites on the St. Lawrence River upstream and downstream from the island of Montreal are described from study of histological sections of individual fish. Community membership included Myxobolus sp. A (intracellular in striated muscle fibers), Myxobolus sp. B (intracellular in striated muscle fibers), Myxobolus sp. C (brain), and Thelohanellus notatus (Mavor, 1916) Kudo, 1929 (loose connective tissue), all of which are histozoic myxobolids displaying strict tissue specificity for trophozoite development. Mean infracommunity richness in fish at the separate localities was estimated to be 0.4 +/- 0.5 to 1.3 +/- 0.7, with a maximum richness of 3 in any 1 fish. Component community richness in host samples was 2 to 4. It is argued that these are relatively high levels of diversity for freshwater fish parasites but that the values are probably conservative because of the study of only portions of individual fish. The percentage of fish infected with myxozoans of any species and infracommunity richness was significantly greater below the island of Montreal compared with above it. It is suspected that increased oligochaete populations at these sites, resulting from sewage-caused organic enrichment of sediments, may have accounted for the observed increased prevalence of infection.