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.

Effects of agricultural pesticides on the health of Rana pipiens frogs sampled from the field.

There is evidence that over the last 30 years, there have been mass declines in diverse geographic locations among amphibian populations due to disease outbreaks. Multiple causes have been suggested to explain this increase in disease incidence. Among these, climate changes, environmental pollution and reduced water quality are gaining attention. Indeed, some chemicals of environmental concerns are known to alter the immune system. It is possible that exposure to these pollutants could alter the immune system of anurans and render them more susceptible to different pathogens. In this study, we sampled Rana pipiens in five different sites near St. Lawrence River (Quebec, Canada) during the months of July and September in 2001. Two of these sites were protected areas, in which low levels of pesticides were detected, while the remaining three sites were located in areas with intensive corn and soybeans cultivations. Our results demonstrated that frogs living in agricultural regions are smaller in size and weight than frogs living in areas with lower levels of pesticides at both sampling times. Moreover, we have observed a significant decrease in the number of splenocytes (cellularity) and the phagocytic activity in frogs sampled in impacted sites. Taken together, these results suggest that frogs living in agricultural regions might be more vulnerable to infections and diseases through their smaller size and alteration of their immune system. Our results also contribute to the overall discussion on factors involved in amphibian declines.

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.

The effects of a primary-treated municipal effluent on the immune system of rainbow trout (Oncorhynchus mykiss): exposure duration and contribution of suspended particles.

Municipal sewage effluents are complex mixtures of contaminants known to disrupt both immune and endocrine functions in aquatic organisms. The present study sought to determine the impacts of municipal effluent on the immune systems of juvenile rainbow trout (Oncorhynchus mykiss), by exposing specimens to low concentrations (0.01%, 0.1%, 1% or 10%) of sewage effluent for periods of 28 or 90 days. The soluble and insoluble fractions of the effluent were also studied to assess the contribution of fractions rich in microorganisms and particles on fish immune systems. To this end, the trout were also exposed to soluble and insoluble fractions of the effluent for a period of 28 days. Immunocompetence was assessed by the following three parameters: phagocytosis, natural cytotoxic cells (NCC) and blastogenesis of lymphocytes under mitogen stimulation. Fish exposed to the 1% sewage effluent concentration for 28 days had enhanced phagocytic activity; at 90 days, phagocytic activity was reduced. T and B lymphocyte proliferation in fish from both groups was similarly stimulated. Phagocytosis and NCC activities were influenced more by the insoluble fraction than the soluble fraction of the effluent. Conversely, mitogen-stimulated T lymphocyte proliferation was enhanced in cells of fish exposed to the soluble fraction of the effluents, with a dampening effect on the insoluble (particulate) fraction of the effluent. In conclusion, the effects of the effluent and its fractions were higher at the cellular-mediated immunity level than at the acquired immunity level. Immunotoxicity data on the soluble fraction of the effluent were more closely associated to data on the unfractionated effluent, but the contribution of the particulate fraction could not be completely ignored for phagocytosis and B lymphocyte proliferation.

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.

Impacts of agriculture on the parasite communities of northern leopard frogs (Rana pipiens) in southern Quebec, Canada.

Given that numerous amphibians are suffering population declines, it is becoming increasingly important to examine the relationship between disease and environmental disturbance. Indeed, while many studies relate anthropogenic activity to changes in the parasitism of snails and fishes, little is known of the impact on the parasites of amphibians, particularly from agriculture. For 2 years, the parasite communities of metamorphic northern leopard frogs from 7 agricultural wetlands were compared with those from 2 reference wetlands to study differences in parasite community diversity and abundance of various species under pristine conditions and 3 categories of disturbance: only agricultural landscape, only pesticides, and agricultural landscape with pesticides. Agricultural (and urban) area was negatively related to species richness, and associated with the near absence of adult parasites and species that infect birds or mammals. We suggest that agriculture and urbanization may hinder parasite transmission to frogs by limiting access of other vertebrate hosts of their parasites to wetlands. The only parasite found at all localities was an unidentified echinostome infecting the kidneys. This parasite dominated communities in localities surrounded by the most agricultural land, suggesting generalist parasites may persist in disrupted habitats. Community composition was associated with dissolved organic carbon and conductivity, but few links were found with pesticides. Pollution effects may be masked by a strong impact of land use on parasite transmission.

Concentration-dependent effects of waterborne zinc on population dynamics of Gyrodactylus turnbulli (Monogenea) on isolated guppies ( Poecilia reticulata ).

The effect of waterborne zinc (Zn) on Gyrodactylus population dynamics was studied on isolated guppies maintained at concentrations ranging from 0 to 240 microg Zn/l. After 1 week pre-exposure to Zn, each fish was experimentally infected with 3 gyrodactylids and parasite numbers were recorded daily on each fish until the fish either died or recovered from infection. Parasite establishment was most successful at 0 and 240 microg Zn/l (97%) compared with the intermediate Zn concentrations. Low to moderate concentrations of Zn were beneficial to the parasite, as evidenced by the concentration-dependent increase in peak parasite burden on recovered fish up to 120 microg Zn/l. In contrast, 240 microg Zn/l may have been toxic to the parasite, as both peak parasite burden (in fish that recovered from infection), and maximum rate of increase of the parasite population (in fish that died) declined at this concentration. The combined effect of infection and Zn is harmful to the fish, because mortality of infected fish (but not uninfected fish) increased with increasing Zn concentrations. We suggest that the observed mortality occurs because of the inability of fish to continuously produce mucous that is a key factor for protecting fish from both waterborne Zn and ectoparasites.

Occurrence of Proteocephalus tetrastomus (Rudolphi, 1810) (Cestoda: Proteocephalidea) in larval rainbow smelt (Osmerus mordax) in North America: identification of a potential pathogen confirmed.

A morphological evaluation and genetic analysis (sequencing of ITS2 region of rDNA) of proteocephalidean cestodes from rainbow smelt (Osmerus mordax) in the Saint Lawrence Estuary, Canada, has shown their conspecificity with Proteocephalus tetrastomus, a specific parasite of smelt (Osmeridae), previously known only from northern Europe, Russia, and Japan. The parasite occurs only in larval, but not adult, smelt in the Saint Lawrence Estuary. Prevalence of larval smelt infection was 42% (n = 50), mean intensity 2.1 +/- 2.4 and mean abundance 1.1 +/- 1.0.

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.

Exposure of leopard frogs to a pesticide mixture affects life history characteristics of the lungworm Rhabdias ranae.

We tested the hypothesis that exposure of leopard frogs ( Rana pipiens) to agricultural pesticides can affect the infection dynamics of a common parasite of ranid frogs, the lungworm Rhabdias ranae. After a 21-day exposure to sublethal concentrations of a pesticide mixture composed of atrazine, metribuzin, aldicarb, endosulfan, lindane and dieldrin, or to control solutions (water, dimethyl sulfoxide), parasite-free juvenile frogs were challenged with 30 infective larvae of R. ranae. Approximately 75% of the larvae penetrated the skin and survived in both exposed and control animals, suggesting that pesticides did not influence host recognition or penetration components of the transmission process. Rather, we found that the migration of R. ranae was significantly accelerated in hosts exposed to the highest concentrations of pesticides, leading to the establishment of twice as many adult worms in the lungs of frogs 21 days post-infection. Pesticide treatment did not influence the growth of lungworms but our results indicate that they matured and reproduced earlier in pesticide-exposed frogs compared to control animals. Such alterations in life history characteristics that enhance parasite transmission may lead to an increase in virulence. Supporting evidence shows that certain components of the frog immune response were significantly suppressed after exposure to the pesticide mixture. This suggests that the immune system of anurans exerts a control over lungworm migration and maturation and that agricultural contaminants can interfere with these control mechanisms. Our results also contribute to the ongoing debate regarding the role that anthropogenic factors could play in the perplexing disease-related die-offs of amphibians observed in several parts of the world.