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Several factors influence whether an organism remains in its local habitat. Parasites can, for example, influence host movement by impacting their behavior, physiology, and morphology. In rivers, fish that swim efficiently against the current are able to maintain their position without being displaced downstream, a behavior referred to as positive rheotaxis. We hypothesized that both the presence and number of ectoparasites on a host would affect the ability of fish to avoid downstream displacement and thus prevent them from remaining in their habitat. We used the guppy-Gyrodactylus host-ectoparasite model to test whether parasite presence and parasite load had an effect on fish rheotaxis. We quantified rheotaxis of sham-infected and parasite-infected fish in a circular flow tank in the laboratory prior to infection and 5-6 days postinfection. Both parasite-infected and sham-infected individuals expressed similar levels of positive rheotaxis prior to infection and after infection. However, with increasing parasite numbers, guppies covered less distance in the upstream direction and spent more time in slower flow zones. These results suggest that higher numbers of Gyrodactylus ectoparasites negatively influence rheotactic movements. Further research is needed to understand the ecological and evolutionary implications of this ectoparasite on fish movement., (© 2024 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

Tetrahymena piriformis belongs to the ciliated protists (ciliates), causing severe economic losses in aquaculture. Chemical drugs currently used usually have toxic side effects, and there is no specific drug against Tetrahymena. Therefore, it is an urgent need to identify new antiparasitic lead compounds. In the present study, the in vitro parasiticidal activity of ethyl acetate (EtOAc) extracts and water extracts from 22 selected traditional Chinese medicines (TCMs) were evaluated against T. piriformis. The EtOAc extract of P. corylifolia turned out to be the most active with the minimum parasiticidal concentration of 100 mg/L within 3 h. Thus, it was separated into 12 fractions by the first-dimensional (D1) normal phase liquid chromatography (NPLC), meanwhile combining with in vitro antiparasitic tests for activity tracking. Subsequently, 8 flavonoids were identified in the active fractions by the second-dimensional (D2) reverse phase liquid chromatography (RPLC) tandem high-resolution mass spectrometry. According to the results, 5 flavonoids were selected for in vitro antiparasitic test, of which isobavachalcone showed the minimum parasiticidal concentration of 3.125 mg/L in 2 h. Bathing treatment of infected guppies with isobavachalcone could significantly reduce the burden of T. piriformis, obtaining a 24-h median effective concentration (24-h EC 50 ) value of 1.916 mg/L. And the concentration of isobavachalcone causing guppies to die within 24 h is 39 times than that of 24-h EC 50 . The results demonstrated that isobavachalcone has the potential to be developed into a novel commercial fish drug against T. piriformis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Behavioural and physiological resistance are key to slowing epidemic spread. We explore the evolutionary and epidemic consequences of their different costs for the evolution of tolerance that trades off with resistance. Behavioural resistance affects social cohesion, with associated group-level costs, while the cost of physiological resistance accrues only to the individual. Further, resistance, and the associated reduction in transmission, benefit susceptible hosts directly, whereas infected hosts only benefit indirectly, by reducing transmission to kin. We therefore model the coevolution of transmission-reducing resistance expressed in susceptible hosts with resistance expressed in infected hosts, as a function of kin association, and analyse the effect on population-level outcomes. Using parameter values for guppies, Poecilia reticulata , and their gyrodactylid parasites, we find that: (1) either susceptible or infected hosts should invest heavily in resistance, but not both; (2) kin association drives investment in physiological resistance more strongly than in behavioural resistance; and (3) even weak levels of kin association can favour altruistic infected hosts that invest heavily in resistance (versus selfish tolerance), eliminating parasites. Overall, our finding that weak kin association affects the coevolution of infected and susceptible investment in both behavioural and physiological resistance suggests that kin selection may affect disease dynamics across systems.

Schyzocotyle acheilognathi is a fish tapeworm native to Asia but has been reported as an alien species on practically all other continents. Its invasive potential is due to its low host specificity and high adaptability to different environments, and its spread to new areas can result in economic and ecological impacts. Studies reporting this species in South America are still scarce, indicating the need to monitor its dispersion to new areas. Herein, tapeworms found in guppies, Poecilia reticulata, from an urban stream located in Belo Horizonte, Minas Gerais, Brazil, in April 2021 were subjected to morphological and molecular characterization. As a result, 5/13 (38.5%) of the P. reticulata specimens evaluated were infected with intestinal tapeworms. It was verified a mean intensity of infection of 7.8 (1-25) and a mean abundance of infection of 3 (0-25). The morphology of the cestodes obtained was compatible with that of S. acheilognathi. Moreover, genetic analysis based on cytochrome c oxidase subunit 1 gene (Cox-1) revealed 97.89-99.77% similarity to isolates of this species from different localities. The possibility that S. acheilognathi is expanding to new regions of South America is discussed.

Cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) is a distinctive member of the serine–threonine protein AGC kinase family and an effective kinase for cAMP signal transduction. In recent years, scuticociliate has caused a lot of losses in domestic fishery farming, therefore, we have carried out morphological and molecular biological studies. In this study, diseased guppies ( Poecilia reticulata ) were collected from an ornamental fish market, and scuticociliate Philaster apodigitiformis Miao et al ., 2009 was isolated. In our prior transcriptome sequencing research, we discovered significant expression of the β-PKA gene in P. apodigitiformis during its infection process, leading us to speculate its involvement in pathogenesis. A complete sequence of the β-PKA gene was cloned, and quantified by quantitative reverse transcription-polymerase chain reaction to analyse or to evaluate the functional characteristics of the β-PKA gene. Morphological identification and phylogenetic analysis based on small subunit rRNA sequence, infection experiments and haematoxylin–eosin staining method were also carried out, in order to study the pathological characteristics and infection mechanism of scuticociliate. The present results showed that: (1) our results revealed that β-PKA is a crucial gene involved in P. apodigitiformis infection in guppies, and the findings provide valuable insights for future studies on scuticociliatosis; (2) we characterized a complete gene, β-PKA , that is generally expressed in parasitic organisms during infection stage and (3) the present study indicates that PKA plays a critical role in scuticociliate when infection occurs by controlling essential steps such as cell growth, development and regulating the activity of the sensory body structures and the irritability system.

Circadian rhythms are timekeeping mechanisms responsible for an array of biological processes. Disruption of such cycles can detrimentally affect animal health. Circadian rhythms are critical in the co-evolution of host–parasite systems, as synchronization of parasite rhythms to the host can influence infection dynamics and transmission potential. This study examines the circadian rhythms in behaviour and activity of a model fish species ( Poecilia reticulata ) in isolation and in shoals, both when uninfected and infected with an ectoparasite ( Gyrodactylus turnbulli ). Additionally, the rhythmical variance of parasite activity under different light conditions as well as rhythmical variance in parasite transmissibility was explored. Overall, infection alters the circadian rhythm of fish, causing nocturnal restlessness. Increased activity of gyrodactylids on the host's skin at night could potentially contribute to this elevated host activity. Whilst migration of gyrodactylids across the host's skin may have caused irritation to the host resulting in nocturnal restlessness, the disruption in guppy activity rhythm caused by the expression of host innate immunity cannot be excluded. We discuss the wider repercussions such behavioural responses to infection have for host health, the implications for animal behaviour studies of diurnal species as well as the application of chronotherapeutic approaches to aquaculture.

The 'good genes' hypothesis for the evolution of male secondary sexual traits poses that female preferences for such traits are driven by indirect genetic benefits. However, support for the hypothesis remains ambiguous, and, in particular, the genetic basis for the benefits has rarely been investigated. Here, we use seminatural populations of Trinidadian guppies to investigate whether sexually selected traits (orange, black and iridescent colouration, gonopodium length and body size) predict fitness measured as the number of grandoffspring, a metric that integrates across fitness components and sexes. Furthermore, we tested whether two potential sources of genetic benefits-major histocompatibility complex (MHC) genotypes and multilocus heterozygosity (MLH)-are significant predictors of fitness and of the size of sexually selected traits. We found a significant, nonlinear effect of the area of black pigmentation and male body size on the number of grandoffspring, suggesting stabilizing selection on black area, and nonlinear selection favouring small body size. MLH was heritable (h 2  = 0.14) and significantly predicted the number of grandoffspring, indicating the potential for genetic benefits based on heterozygosity. We also found support for local heterozygosity effects, which may reflect a noneven distribution of genetic load across the genome. MHC genotype was not significantly associated with any tested fitness component, or with the load of Gyrodactylus parasites. Neither MHC nor MLH was significant predictor of sexually selected traits. Overall, our results highlight the role of heterozygosity in determining fitness, but do not provide support for male sexually selected traits being indicators of genetic quality., (© 2023 John Wiley & Sons Ltd.)

The reliance on chemical communication is well established for evading predation in aquatic systems. Only a few studies have found evidence that chemical cues released from aquatic animals infected with parasites alter behaviour. Furthermore, the link between putative chemical cues and susceptibility to infection has not been studied. The objectives of this study were to determine if exposure to chemical cues from Gyrodactylus turnbulli -infected guppies ( Poecilia reticulata ) at various times post-infection resulted in altered behaviour of uninfected conspecifics, and if prior exposure to this putative infection cue reduced transmission. Guppies responded to this chemical cue. Those exposed for 10 min to cues released from fish that had been infected for 8 or 16 days spent less time in the centre half of the tank. Continuous exposure to infection cues for 16 days did not alter guppy shoal behaviour but provided partial protection against infection when the parasite was introduced. Shoals exposed to these putative infection cues became infected, but infection intensity increased more slowly and to a lower peak compared with shoals exposed to the control cue. These results indicate that guppies show subtle behavioural responses to infection cues, and that exposure to infection cues reduces the intensity of outbreaks.

Background: Mathematical modelling of host-parasite systems has seen tremendous developments and broad applications in theoretical and applied ecology. The current study focuses on the infection dynamics of a gyrodactylid-fish system. Previous experimental studies have explored the infrapopulation dynamics of co-infecting ectoparasites, Gyrodactylus turnbulli and G. bullatarudis, on their fish host, Poecilia reticulata, but questions remain about parasite microhabitat preferences, host survival and parasite virulence over time. Here, we use more advanced statistics and a sophisticated mathematical model to investigate these questions based on empirical data to add to our understanding of this gyrodactylid-fish system., Methods: A rank-based multivariate Kruskal-Wallis test coupled with its post-hoc tests and graphical summaries were used to investigate the spatial and temporal parasite distribution of different gyrodactylid strains across different host populations. By adapting a multi-state Markov model that extends the standard survival models, we improved previous estimates of survival probabilities. Finally, we quantified parasite virulence of three different strains as a function of host mortality and recovery across different fish stocks and sexes., Results: We confirmed that the captive-bred G. turnbulli and wild G. bullatarudis strains preferred the caudal and rostral regions respectively across different fish stocks; however, the wild G. turnbulli strain changed microhabitat preference over time, indicating microhabitat preference of gyrodactylids is host and time dependent. The average time of host infection before recovery or death was between 6 and 14 days. For this gyrodactylid-fish system, a longer period of host infection led to a higher chance of host recovery. Parasite-related mortalities are host, sex and time dependent, whereas fish size is confirmed to be the key determinant of host recovery., Conclusion: From existing empirical data, we provided new insights into the gyrodactylid-fish system. This study could inform the modelling of other host-parasite interactions where the entire infection history of the host is of interest by adapting multi-state Markov models. Such models are under-utilised in parasitological studies and could be expanded to estimate relevant epidemiological traits concerning parasite virulence and host survival., (© 2022. The Author(s).)

The major histocompatibility complex (MHC) plays an important role in infectious disease resistance. The presence of certain MHC alleles and functionally similar groups of MHC alleles (i.e., supertypes) has been associated with resistance to particular parasite species. Farmed and domesticated fish stocks are often depleted in their MHC alleles and supertype diversity, possibly as a consequence of artificial selection for desirable traits, inbreeding (loss of heterozygosity), genetic drift (loss of allelic diversity) and/or reduced parasite biodiversity. Here we quantify the effects of depletion of MHC class II genotype and supertype variation on resistance to the parasite Gyrodactylus turnbulli in guppies (Poecilia reticulata). Compared to the descendants of wild-caught guppies, ornamental fish had a significantly reduced MHC variation (i.e., the numbers of MHC alleles and supertypes per individual, and per population). In addition, ornamental fish were significantly more susceptible to G. turnbulli infections, accumulating peak intensity 10 times higher than that of their wildtype counterparts. Four out of 13 supertypes were associated with a significantly reduced parasite load, and the presence of some supertypes had a dramatic effect on the intensity of infection. Remarkably, the ornamental and wildtype fish differed in the supertypes that were associated with parasite resistance. Analysis with a genetic algorithm showed that resistance-conferring supertypes of the wildtype and ornamental fish shared two unique amino acids in the peptide-binding region of the MHC that were not found in any other alleles. These data show that the supertype demarcation captures some, but not all, of the variation in the immune function of the alleles. This study highlights the importance of managing functional MHC diversity in livestock, and suggests there might be some immunological redundancy among MHC supertypes., (© 2020 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

The fishborne zoonotic trematode Haplorchis pumilio (Looss, 1896) is here reported for the first time in Brazil based on morphological, experimental and molecular studies. Pleurolophocercous cercariae emerged from the invasive snail Melanoides tuberculata collected in the municipality of Ceará-Mirim, state of Rio Grande do Norte, Northeast region of Brazil, in November 2018, were used for experimental infection of guppies, Poecilia reticulata. Metacercariae were extracted from the caudal musculature of these fishes and were used to infect mice. Adult parasites recovered in the small intestine of the mice were morphologically identified as H. pumilio. Molecular sequences were obtained for the Brazilian and Peruvian isolates of H. pumilio, and were compared with data available in GenBank. Analyses of fragments of the nuclear genes 28S (1219 bp) and ITS-2 (290 bp) revealed 98.48-100% similarity between the South American and Asian isolates of H. pumilio. Moreover, new sequences of the mitochondrial gene cox-1 obtained for the Brazilian (797 bp) and Peruvian (646 pb) isolates were 100% similar to a Mexican isolate of this species and 97.54% similar to an isolate from Thailand. This finding reveals the potential for occurrence of human haplorchiasis in Brazil, especially because of the increasing popularity of raw fish dishes in the country., (Copyright © 2020 Elsevier B.V. All rights reserved.)

AbstractSexually selected ornaments range from highly dynamic traits to those that are fixed during development and relatively static throughout sexual maturity. Ornaments along this continuum differ in the information they provide about the qualities of potential mates, such as their parasite resistance. Dynamic ornaments enable real-time assessment of the bearer's condition: they can reflect an individual's current infection status, or they can reflect resistance to recent infections. Static ornaments, however, are not affected by recent infection but may instead indicate an individual's genetically determined resistance, even in the absence of infection. Given the typically aggregated distribution of parasites among hosts, infection is unlikely to affect the ornaments of the vast majority of individuals in a population: static ornaments may therefore be the more reliable indicators of parasite resistance. To test this hypothesis, we quantified the ornaments of male guppies ( Poecilia reticulata ) before experimentally infecting them with Gyrodactylus turnbulli . Males with more left-right symmetrical black coloration and those with larger areas of orange coloration, both static ornaments, were more resistant. However, males with more saturated orange coloration, a dynamic ornament, were less resistant. Female guppies often prefer symmetrical males with larger orange ornaments, suggesting that parasite-mediated natural and sexual selection act in concert on these traits.

The global movement of pathogens is altering populations and communities through a variety of direct and indirect ecological pathways. The direct effect of a pathogen on a host is reduced survival, which can lead to decreased population densities. However, theory also suggests that increased mortality can lead to no change or even increases in the density of the host. This paradoxical result can occur in a regulated population when the pathogen's negative effect on survival is countered by increased reproduction at the lower density. Here, we analyze data from a long-term capture-mark-recapture experiment of Trinidadian guppies ( Poecilia reticulata ) that were recently infected with a nematode parasite ( Camallanus cotti ). By comparing the newly infected population with a control population that was not infected, we show that decreases in the density of the infected guppy population were transient. The guppy population compensated for the decreased survival by a density-dependent increase in recruitment of new individuals into the population, without any change in the underlying recruitment function. Increased recruitment was related to an increase in the somatic growth of uninfected fish. Twenty months into the new invasion, the population had fully recovered to preinvasion densities even though the prevalence of infection of fish in the population remained high (72%). These results show that density-mediated indirect effects of novel parasites can be positive, not negative, which makes it difficult to extrapolate to how pathogens will affect species interactions in communities. We discuss possible hypotheses for the rapid recovery., Competing Interests: The authors declare no competing interest.

In accordance with the Red Queen hypothesis, the lower genotypic diversity in clonally reproducing species should make them easier targets for pathogen infection, especially when closely related sexually reproducing species occur in close proximity. We analyzed two populations of clonal P. formosa and their sexual parental species P. mexicana by correlating individual parasite infection with overall and immune genotype. Our study revealed lower levels of overall genotypic diversity and marginally fewer MHC class I alleles in P. formosa individuals compared to sexually reproducing P. mexicana. Parasite load, however, differed only between field sites but not between species. We hypothesize that this might be due to slightly higher genotypic diversity in P. formosa at the innate immune system (toll like receptor 8) which is likely due to the species' hybrid origin. In consequence, it appears that clonal individuals do not necessarily suffer a disadvantage compared to sexual individuals when fighting parasite infection., Competing Interests: The authors have declared that no competing interests exist.

Treated wastewater (TWW) constitutes a sustainable water resource and has been used for fish culture in some countries around the world, although there are no comprehensive data on the effect of TWW on fish growth and health in the context of aquaculture production. Our objectives were to examine how fish culture in TWW affected fish growth and fitness, as well as compliance with the international standards for safe consumption. Guppy (Poecilia reticulata) fingerlings were reared in 0%, 50% and 100% tertiary TWW (TTWW), from the age of five days, for a period of four months. In water analyses, 33 out of 67 tested organic micropollutants (OMPs) were detected in the TTWW samples at least once, at concentrations that are typically reported in domestic TTWW. Fish survival ranged between 77-80% and did not differ between treatment groups. Fish growth and mortality following challenge infection with Tetrahymena sp. (which ranged between 64-68%), were similar among treatment groups. Of tested immunological parameters, lysozyme and anti-protease was similar among treatments while complement activity was highest in the 50% TTWW-reared fish. No abnormalities were observed in the histopathological analysis. Levels of heavy metals, polychlorinated-biphenyls (PCBs) and organochlorines (OCs) in fish were below the detection limit and below the Food and Agriculture Organization of the United Nations (FAO) and the European Union EU maximal permitted levels in food fish. Results suggest that the yield of fish grown in TTWW is potentially similar to that in freshwater, and the produced fish comply with the standards of consumer safety. The results are in line with previous studies that examined the feasibility of TWW-fed aquaculture., Competing Interests: The authors have declared that no competing interests exist.

Species of the genus Posthodiplostomum are intestinal parasites of piscivorous birds with worldwide distribution. Recent molecular surveys have focused on developmental stages from vertebrate hosts, with few sequences from larvae found in molluscs. Moreover, most published molecular data are from collections in North America, Europe and Asia, and there are no data for South American species. In the present study, cercariae found in the freshwater limpet Gundlachia ticaga from Brazil were used for morphological, experimental and molecular studies. The longifurcate cercariae, Neascus-type metacercariae obtained in experimentally infected guppies (Poecilia reticulata), and an adult parasite recovered from a mouse were morphologically identified as Posthodiplostomum nanum. Sequences of DNA from internal transcribed spacer (ITS) rDNA and cytochrome c oxidase 1 (cox1) from the cercariae and metacercariae were compared with published data, yielding no matches (ITS divergence ≥ 5.5%, cox1 ≥ 13.9%). Phylogenetic analyses of the ITS sequences suggest paraphyly within the genus Posthodiplostomum, or misidentifications of parasites in molecular studies. For example, ITS sequences of Mesoophorodiplostomum pricei and Posthodiplostomum centrarchi or the unnamed species Posthodiplostomum sp. 8 diverge by only 1.1-1.2% and 0.6%, respectively, and all three species fall within a well-supported clade, suggesting that these isolates are congeneric. The phylogenetic tree obtained for cox1 sequences, although not well resolved, reveals that the type species of the genus, Posthodiplostomum cuticola, does not group with any species previously identified as Posthodiplostomum. Overall, the analyses of molecular data here obtained for P. nanum compared with sequences of related species suggest that a review of this group is necessary. Such studies may result in a split of the genus Posthodiplostomum and the transference of some species currently assigned in this genus to other already described genera., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Two cryptic species of haploporid digeneans belonging in Saccocoelioides are described from Costa Rica: one from a poeciliid fish, Poecilia gillii, and the other from a characid fish, Astyanax aeneus. Traditional morphological features are examined and found to be largely inefficient for differentiating among the new species and 20 New World congeners. Comparison of ribosomal DNA sequences among the 2 new species, 2 North American, 4 Middle American, and 3 South American species of Saccocoelioides, including the type-species Saccocoelioides nanii from Argentina and Saccocoelioides sogandaresi from Texas, is effective in differentiating among these species. A Bayesian inference analysis is conducted using a concatenated alignment of the same 2 ribosomal gene regions from 35 species belonging in the Haploporoidea and rooted by the atractotrematid Isorchis anomalus. The analysis provides stronger support for a close relationship between species in Saccocoelioides and Intromugil; thus, Intromugil is transferred from the Waretrematinae to the Chalcinotrematinae. Additionally, interrelationships among 11 species of Saccocoelioides are resolved and reveal a distinct genetic rift suggesting the presence of 2 distinct lineages within the genus, 1 containing the type-species and the other containing a group of species that more closely resemble other genera in the subfamily. Taxonomy of Saccocoelioides is discussed, and observations made during the present study justify taking several taxonomic actions: new combinations are proposed for Saccocoelioides tilapiae n. comb., formerly in the now-defunct Culuwiya, and Saccocoelioides ruedasueltensis n. comb., formerly in Chalcinotrema; Saccocoelioides guaporense nomen novem is proposed for Lecithobotrioides elongatus; Saccocoelioides papernai is considered a junior subjective synonym of Saccocoelioides overstreeti; Saccocoelioides godoyi is considered a junior subjective synonym of Saccocoelioides szidati; Saccocoelioides magnorchis and Saccocoelioides saccodontis are considered species inquirendae; and Saccocoelioides adelae is considered a nomen nudum. Four new hosts are reported for Saccocoelioides cichlidorum: Amphilophus lyonsi, Amatitlania nigrofasciatus, Amatitlania septemfasciatus, and Hypsophrys nicaraguensis. Molecular data call into question some existing species identifications in Saccocoelioides and reveal that molecular tools combined with traditional taxonomy are required for accurately identifying species in the genus. Twenty-two species of Saccocoelioides are formally accepted, but it is noted that as new molecular data become available, some of these species may be transferred to other genera in the subfamily.

While group formation provides antipredatory defences, increases foraging efficiency and mating opportunities, it can be counterintuitive by promoting disease transmission amongst social hosts. Upon introduction of a pathogen, uninfected individuals often modify their social preferences to reduce infection risk. Infected hosts also exhibit behavioural changes, for example, removing themselves from a group to prevent an epidemic. Conversely, here we show how Trinidadian guppies infected with a directly transmitted ectoparasite, Gyrodactylus turnbulli, significantly increase their contact rates with uninfected conspecifics. As uninfected fish never perform this behaviour, this is suggestive of a parasite-mediated behavioural response of infected hosts, presumably to offload their parasites. In the early stages of infection, however, such behavioural modifications are ineffective in alleviating parasite burdens. Additionally, we show that fish exposed to G. turnbulli infections for a second time, spent less time associating than those exposed to parasites for the first time. We speculate that individuals build and retain an infection cue repertoire, enabling them to rapidly recognize and avoid infectious conspecifics. This study highlights the importance of considering host behavioural modifications when investigating disease transmission dynamics.

The ornamental fish trade provides a pathway for the global translocation of aquatic parasites. We examined a total of 1020 fish imported from Singapore, Malaysia, Thailand, or Sri Lanka to Australia (including freshwater and marine fish species) for monogenean ectoparasites. Fish were received following veterinary certification that they showed no clinical signs of pests and diseases from the exporting country and visual inspection at Australian border control. Australian import conditions require mandatory treatment for goldfish with parasiticides (e.g. trichlorfon, formaldehyde, sodium chloride) for the presence of gill flukes (Dactylogyrus vastator Nybelin, 1924 and Dactylogyrus extensus Mueller and Van Cleave, 1932) prior to export. Over 950 individual parasites were detected in five imported fish species, representing 14 monogenean species. Seven Dactylogyrus spp. including D. vastator and three Gyrodactylus spp. infected goldfish, Carassius auratus Linnaeus, 1758, from Malaysia, Singapore, and Thailand. Dactylogyrus ostraviensis Řehulka, 1988, infected rosy barb, Pethia conchonius Hamilton, 1822, from Singapore, Sri Lanka, and Thailand while two Trianchoratus spp. infected three spot gourami, Trichopodus trichopterus Pallas, 1970 and pearl gourami Trichopodus leerii Bleeker, 1852, from Sri Lanka. Urocleidoides reticulatus Mizelle & Price, 1964, infected guppy, Poecilia reticulata Peters, 1859, from Sri Lanka. The discovery of D. vastator in goldfish, as well as 13 other monogenean species, shows that pre-export health requirements, which include chemical treatment of goldfish, and inspection of all ornamental fish species did not prevent infection by monogeneans. Inspection prior to exportation and at border control must account for the highly cryptic nature of monogenean parasites and consider alternatives to current pre-export conditions and visual inspection at border control.

Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.

The major histocompatibility complex (MHC) is crucial to the adaptive immune response of vertebrates and is among the most polymorphic gene families known. Its high diversity is usually attributed to selection imposed by fast-evolving pathogens. Pathogens are thought to evolve to escape recognition by common immune alleles, and, hence, novel MHC alleles, introduced through mutation, recombination, or gene flow, are predicted to give hosts superior resistance. Although this theoretical prediction underpins host-pathogen "Red Queen" coevolution, it has not been demonstrated in the context of natural MHC diversity. Here, we experimentally tested whether novel MHC variants (both alleles and functional "supertypes") increased resistance of guppies ( Poecilia reticulata ) to a common ectoparasite ( Gyrodactylus turnbulli ). We used exposure-controlled infection trials with wild-sourced parasites, and Gyrodactylus -naïve host fish that were F 2 descendants of crossed wild populations. Hosts carrying MHC variants (alleles or supertypes) that were new to a given parasite population experienced a 35-37% reduction in infection intensity, but the number of MHC variants carried by an individual, analogous to heterozygosity in single-locus systems, was not a significant predictor. Our results provide direct evidence of novel MHC variant advantage, confirming a fundamental mechanism underpinning the exceptional polymorphism of this gene family and highlighting the role of immunogenetic novelty in host-pathogen coevolution., Competing Interests: The authors declare no conflict of interest.

Introduced alien fish species and their associated parasites may result in a serious threat to indigenous biodiversity. Furthermore, this may have negative impacts on cultured fish as well as on native parasitic fauna. In the present study, the invasive Asian nematode, Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae), is reported from the guppy (Poecilia reticulata) for the first time in Africa. This parasite is assumed to be introduced into Africa along with the introduction of exotic poeciliid fishes, which are known to be the most common hosts of C. cotti in ornamental fish industry worldwide.The presence of this parasite in both aquarium-cultured fish as well as fish from natural waterbodies is evidence of the introduction of the alien organisms due to insufficient prophylactic veterinary control during transfer of non-native hosts between countries and the spread of them by the anthropogenic introduction to natural systems.

As part of ongoing surveys of the gyrodactylid parasite fauna of freshwater fishes in Mexico, we recorded the infection of three species of poeciliids (Poecilia mexicana, Poeciliopsis gracilis, and Pseudoxiphophorus bimaculatus [syn.=Heterandria bimaculata]) with Gyrodactylus cichlidarum, a monogenean parasite of cichlid fishes, which has been co-introduced globally with its translocated, African "tilapia" hosts. This tilapia pathogen was found on poeciliid fishes both within their native distribution range in the Gulf of Mexico slope, as well as on invasive species artificially introduced to the Mexican highlands, to rivers draining into the Pacific Ocean. Identity of G. cichlidarum was confirmed by morphological and molecular analyses. Prevalence and abundance of infection were low, but this is the first record of G. cichlidarum infecting poeciliids (Cyprinodontiformes), which are distantly related to this parasite's primary cichlid fish hosts (Perciformes). This study provides evidence that G. cichlidarum, a recognized pathogen which has been co-introduced globally with its cichlid fish hosts for aquacultural purposes, is able to infect non-related poeciliid fishes inhabiting water bodies adjacent to tilapia farms, thereby potentially increasing its ability to disperse between farms and different river basins. It is of particular concern that G. cichlidarum was found on poeciliids, as these invasive fishes have been introduced worldwide and could act as carriers for this parasite known to induce significant mortality of farmed tilapias - globally, the second most important freshwater aquaculture fish group, after the carps., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Goodeid topminnows are live-bearing fishes endemic to the Mexican Highlands (Mesa Central, MC). Unfortunately, in the MC, environmental degradation and introduced species have pushed several goodeid species to the brink of extinction. Invasive fishes can introduce exotic parasites, and the most abundant goodeid, blackfin goodea Goodea atripinnis Jordan, is parasitised by six exotic helminths. Poeciliids are widely dispersed invasive fishes, which exert negative ecological effects on goodeids. Poeciliids host several species of the monogenean genus Gyrodactylus von Nordmann, 1832, including pathogenic, invasive parasites. Here, we looked for evidence of Gyrodactylus species switching hosts from poeciliids to goodeids., Methods: Fish were collected in rivers draining the MC into both sides of the continental divide. Hosts were screened for gyrodactylid parasites in localities where G. atripinnis and poeciliids occurred sympatrically. Gyrodactylus specimens were characterised morphologically (attachment apparatus) and molecularly (internal transcribed spacer region, ITS). A Bayesian phylogenetic tree using ITS sequences established relationships between gyrodactylids collected from goodeid fishes and those from parasites infecting poeciliids., Results: Gyrodactylids were collected from G. atripinnis in six localities on both sides of the watershed where exotic poeciliids occurred sympatrically. Morphological and molecular analyses indicated the presence of four undescribed species of Gyrodactylus infecting this goodeid host. Gyrodactylus tomahuac n. sp., the most abundant and geographically widespread species, is described here. The other three Gyrodactylus spp. are not described, but their ITS sequences are used as molecular data presented here, are the only available for gyrodactylids infecting goodeid fishes. Morphological and molecular data suggest that two distinct groups of gyrodactylids infect goodeids, one of which shares a common ancestor with gyrodactylids parasitizing poeciliids., Conclusions: No evidence was found of gyrodactylids switching hosts from invasive poeciliids to endemic goodeids, nor vice versa. Moreover, considering that G. atripinnis is known to host both Gyrodactylus lamothei Mendoza-Palmero, Sereno-Uribe & Salgado-Maldonado, 2009 and Gyrodactylus mexicanus Mendoza-Palmero, Sereno-Uribe & Salgado-Maldonado, 2009, with the addition of G. tomahuac n. sp. and the three undescribed Gyrodactylus spp. reported, at least six gyrodactylids may infect this host. This would make monogeneans the second most abundant parasite group infecting G. atripinnis, which to date is known to harbour 22 helminth species: nine digeneans, five nematodes, four cestodes, three monogeneans and one acanthocephalan.

Inbreeding can threaten population persistence by reducing disease resistance through the accelerated loss of gene diversity (i.e. heterozygosity). Such inbreeding depression can affect many different fitness-related traits, including survival, reproductive success, and parasite susceptibility. Empirically quantifying the effects of inbreeding on parasite resistance is therefore important for ex-situ conservation of vertebrates. The present study evaluates the disease susceptibility of individuals bred under three different breeding regimes (inbred, crossed with full siblings; control, randomly crossed mating; and fully outbred). Specifically, we examined the relationship between inbreeding coefficient (F-coefficient) and susceptibility to Gyrodactylus turnbulli infection in a live bearing vertebrate, the guppy Poecilia reticulata. Host-breeding regime significantly affected the trajectories of parasite population growth on individual fish. Inbred fish showed significantly higher mean parasite intensity than fish from the control and outbred breeding regimes, and in addition, inbred fish were slower in purging their gyrodactylid infections. We discuss the role of inbreeding on the various arms of the immune system, and argue that the increased disease susceptibility of inbred individuals could contribute to the extinction vortex. This is one of the first studies to quantify the effects of inbreeding and breeding regime on disease susceptibility in a captive bred vertebrate of wild origin, and it highlights the risks faced by small (captive-bred) populations when exposed to their native parasites., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Natural enemies such as predators and parasites are known to shape intraspecific variability of behaviour and personality in natural populations, yet several key questions remain: (i) What is the relative importance of predation vs. parasitism in shaping intraspecific variation of behaviour across generations? (ii) What are the contributions of genetic and plastic effects to this behavioural divergence? (iii) And to what extent are responses to predation and parasitism repeatable across independent evolutionary lineages? We addressed these questions using Trinidadian guppies (Poecilia reticulata) (i) varying in their exposure to dangerous fish predators and Gyrodactylus ectoparasites for (ii) both wild-caught F0 and laboratory-reared F2 individuals and coming from (iii) multiple independent evolutionary lineages (i.e. independent drainages). Several key findings emerged. First, a population's history of predation and parasitism influenced behavioural profiles, but to different extent depending on the behaviour considered (activity, shoaling or boldness). Second, we had evidence for some genetic effects of predation regime on behaviour, with differences in activity of F2 laboratory-reared individuals, but not for parasitism, which had only plastic effects on the boldness of wild-caught F0 individuals. Third, the two lineages showed a mixture of parallel and nonparallel responses to predation/parasitism, with parallel responses being stronger for predation than for parasitism and for activity and boldness than for shoaling. These findings suggest that different sets of behaviours provide different pay-offs in alternative predation/parasitism environments and that parasitism has more transient effects in shaping intraspecific variation of behaviour than does predation., (© 2016 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2016 European Society For Evolutionary Biology.)

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.)

Recognizing and associating with specific individuals, such as conspecifics or kin, brings many benefits. One mechanism underlying such recognition is imprinting: the long-term memory of cues encountered during development. Typically, juveniles imprint on cues of nearby individuals and may later associate with phenotypes matching their 'recognition template'. However, phenotype matching could lead to maladaptive social decisions if, for instance, individuals imprint on the cues of conspecifics infected with directly transmitted diseases. To investigate the role of imprinting in the sensory ecology of disease transmission, we exposed juvenile guppies,Poecilia reticulata, to the cues of healthy conspecifics, or to those experiencing disease caused by the directly transmitted parasite Gyrodactylus turnbulli In a dichotomous choice test, adult 'disease-imprinted' guppies preferred to associate with the chemical cues of G. turnbulli-infected conspecifics, whereas 'healthy-imprinted' guppies preferred to associate with cues of uninfected conspecifics. These responses were only observed when stimulus fish were in late infection, suggesting imprinted fish responded to cues of disease, but not of infection alone. We discuss how maladaptive imprinting may promote disease transmission in natural populations of a social host., (© 2016 The Author(s).)

To better understand the spread of disease in nature, it is fundamentally important to have broadly applicable model systems with readily available species which can be replicated and controlled in the laboratory. Here we used an experimental model system of fish hosts and monogenean parasites to determine whether host sex, group size and group composition (single-sex or mixed-sex) influenced host-parasite dynamics at an individual and group level. Parasite populations reached higher densities and persisted longer in groups of fish compared with isolated hosts and reached higher densities on isolated females than on isolated males. However, individual fish within groups had similar burdens to isolated males regardless of sex, indicating that females may benefit more than males by being in a group. Relative condition was positively associated with high parasite loads for isolated males, but not for isolated females or grouped fish. No difference in parasite dynamics between mixed-sex groups and single-sex groups was detected. Overall, these findings suggest that while host sex influences dynamics on isolated fish, individual fish in groups have similar parasite burdens, regardless of sex. We believe our experimental results contribute to a mechanistic understanding of host-parasite dynamics, although we are cautious about directly extrapolating these results to other systems.

We evaluated the extent to which males and females evolve along similar or different trajectories in response to the same environmental shift. Specifically, we used replicate experimental introductions in nature to consider how release from a key parasite (Gyrodactylus) generates similar or different defence evolution in male vs. female guppies (Poecilia reticulata). After 4-8 generations of evolution, guppies were collected from the ancestral (parasite still present) and derived (parasite now absent) populations and bred for two generations in the laboratory to control for nongenetic effects. These F2 guppies were then individually infected with Gyrodactylus, and infection dynamics were monitored on each fish. We found that parasite release in nature led to sex-specific evolutionary responses: males did not show much evolution of resistance, whereas females showed the evolution of increased resistance. Given that male guppies in the ancestral population had greater resistance to Gyrodactylus than did females, evolution in the derived populations led to reduction of sexual dimorphism in resistance. We argue that previous selection for high resistance in males constrained (relative to females) further evolution of the trait. We advocate more experiments considering sex-specific evolutionary responses to environmental change., (© 2015 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2015 European Society For Evolutionary Biology.)

Parasites are detrimental to host fitness and therefore should strongly select for host defence mechanisms. Yet, hosts vary considerably in their observed parasite loads. One notable source of inter-individual variation in parasitism is host sex. Such variation could be caused by the immunomodulatory effects of gonadal steroids. Here we assess the influence of gonadal steroids on the ability of guppies (Poecilia reticulata) to defend themselves against a common and deleterious parasite (Gyrodactylus turnbulli). Adult male guppies underwent 31 days of artificial demasculinization with the androgen receptor-antagonist flutamide, or feminization with a combination of flutamide and the synthetic oestrogen 17 β-estradiol, and their parasite loads were compared over time to untreated males and females. Both demasculinized and feminized male guppies had lower G. turnbulli loads than the untreated males and females, but this effect appeared to be mainly the result of demasculinization, with feminization having no additional measurable effect. Furthermore, demasculinized males, feminized males and untreated females all suffered lower Gyrodactylus-induced mortality than untreated males. Together, these results suggest that androgens reduce the ability of guppies to control parasite loads, and modulate resistance to and survival from infection. We discuss the relevance of these findings for understanding constraints on the evolution of resistance in guppies and other vertebrates.

A common evolutionary response to predation pressure is increased investment in reproduction, ultimately resulting in a fast life history. Theory and comparative studies suggest that short-lived organisms invest less in defence against parasites than those that are longer lived (the pace of life hypothesis). Combining these tenets of evolutionary theory leads to the specific, untested prediction that within species, populations experiencing higher predation pressure invest less in defence against parasites. The Trinidadian guppy, Poecilia reticulata, presents an excellent opportunity to test this prediction: guppy populations in lower courses of rivers experience higher predation pressure, and as a consequence have evolved faster life histories, than those in upper courses. Data from a large-scale field survey showed that fish infected with Gyrodactylus parasites were of a lower body condition (quantified using the scaled mass index) than uninfected fish, but only in lower course populations. Although the evidence we present is correlational, it suggests that upper course guppies sustain lower fitness costs of infection, i.e. are more tolerant, than lower course guppies. The data are therefore consistent with the pace of life hypothesis of parasite defence allocation, and suggest that life-history traits mediate the indirect effect of predators on the parasites of their prey., (© 2015 The Author(s).)

Objective: Lernaeids are crustacean parasites that are globally distributed among freshwater and marine fish. Approximately 110 species of Lernaeids have been divided into 14 genera. The most common species of Lernaeids is Lernaea cyprinacea, which has been transmitted to ornamental fish worldwide. The economic importance of L. cyprinacea is increasing because of the epidemic caused by the parasite in most of the ornamental fish breeding centers in different parts of the world. The parasite affects its host's health, decreases growth rate, and causes abnormal metabolic activity. Accumulation of these parasites in some parts of the body causes painful points and has harmful outcomes for the functioning and survival of the host., Methods: The present study was conducted to examine the prevalence of L. cyprinacea among ornamental fish within 1 year from September-October 2011 to September-October 2012. In total, 3520 fish [3380 mollies (Poecilia latipinna) and 140 swordtails (Xiphophorus helleri)] were collected from 10 fish maintenance and breeding centers in Kerman, Iran., Results: Of 3520 fish, only 186 fish (5.3%) were infected with L. cyprinacea, and the remaining fish (94.7%) were not infected. The swordtails (X. helleri) and mollies (P. latipinna) showed the highest (10.7%) and the lowest (5.1%) level of infection, respectively. In other words, there was a significant correlation between species of fish and L. cyprinacea infection (p<0.05). The highest prevalence (39.4%) and the lowest prevalence (0%) of L. cyprinacea were observed during summer and winter, respectively, which can be attributed to the temperature difference between the two seasons. Also, the histopathologic examination of sections revealed some lesions in the epidermis, dermis, and muscles., Conclusion: Considering the existing L. cyprinacea infection in ornamental fish reproduction and breeding centers in Kerman, public knowledge should increase through management methods; physical and chemical treatments should also be applied to inform the public regarding the risk of infection and other internal diseases that may be associated with ornamental fish.

Hosts and parasites are in a perpetual co-evolutionary "arms race". Due to their short generation time and large reproductive output, parasites are commonly believed to be ahead in this race, although increasing evidence exists that parasites are not always ahead in the arms race - in part owing to evolutionary lineage and recent ecological history. We assess local adaptation of hosts and parasites, and determine whether adaptation was influenced by ecological or evolutionary history, using full reciprocal cross-infections of four Gyrodactylus ectoparasite populations and their four guppy (Poecilia reticulata) host populations in Trinidad. To consider effects of evolutionary lineage and recent ecology, these four populations were collected from two different river drainages (Marianne and Aripo) and two different predation environments (high and low). The highest infection levels were obtained when parasites from the Aripo lineage infected guppies from the Marianne lineage, indicating a higher infectivity, virulence and/or reproductive success of the Aripo parasites. Aripo lineage guppies were also better able to limit Gyrodactylus population growth than guppies from the Marianne River, indicating their strong "resistance" to Gyrodactylus regardless of the source of the parasite. Predation environment had no detectable influence on host-parasite population dynamics of sympatric or allopatric combinations. The much stronger effect of evolutionary lineage (i.e., river) than recent ecological history (i.e., predation) emphasises its importance in driving co-evolutionary dynamics, and should be explored further in future studies on local host-parasite adaptation., (Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

The morphological, biological, and molecular characteristics of Cryptosporidium piscine genotype 1 from the guppy (Poecilia reticulata) are described, and the species name Cryptosporidium huwi n. sp. is proposed to reflect its genetic and biological differences from gastric and intestinal Cryptosporidium species. Oocysts of C.huwi n. sp. over-lap in size with Cryptosporidium molnari, measuring approximately 4.4-4.9 µm (mean 4.6) by 4.0-4.8 µm (mean 4.4 µm) with a length to width ratio of 1.04 (0.92-1.35) (n = 50). Similar to C.molnari, C.huwi n. sp. was identified in the stomach only and clusters of oogonial and sporogonial stages were identified deep within the epithelium. However, phylogenetic analysis of 18S rRNA sequences indicated that C. huwi n. sp. exhibited 8.5-9.2% and 3.5% genetic distance from C.molnari isolates and piscine genotype 7 respectively. At the actin locus, the genetic distance between C.huwi n. sp. and C.molnari was 16.6%. The genetic distance between C.huwi n. sp. and other Cryptosporidium species at the 18S locus was 13.2%-17% and at the actin locus was 18.9%-26.3%. Therefore C. huwi n. sp. is genetically distinct from previously described Cryptosporidium species., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Despite the large number of reports of species of Clinostomum from vertebrate hosts in South America, studies evaluating the molluscan transmitters of these parasites are scarce. In the present study, clinostomatoid cercariae shed from 0.02% (4/17,485) specimens of Biomphalaria spp., collected at the Pampulha reservoir, Belo Horizonte, Minas Gerais, Brazil, were used for experimental infection of Poecilia reticulata . Samples of cercariae from molluscs and metacercariae experimentally obtained from fish were subjected to morphological and molecular analyses and compared with species of Clinostomum reported in the Americas. The cercariae and metacercariae, here identified as Clinostomum sp., present general morphology similar to that reported for Clinostomum marginatum , however, from molecular point of view, differ significantly from North American C. marginatum and other species of Clinostomum reported in South America. These results suggest that the diversity of Clinostomum found in Brazil may be underestimated. Additional studies aimed at molecular characterization of South American species of Clinostomum, including the finding of specimens with sequences similar to that reported for C. marginatum in North America are required.

Predation pressure can alter the morphology, physiology, life history, and behavior of prey; each of these in turn can change how surviving prey interact with parasites. These trait-mediated indirect effects may change in direction or intensity during growth or, in sexually dimorphic species, between the sexes. The Trinidadian guppy, Poecilia reticulata presents a unique opportunity to examine these interactions; its behavioral ecology has been intensively studied in wild populations with well-characterized predator faunas. Predation pressure is known to have driven the evolution of many guppy traits; for example, in high-predation sites, females (but not males) tend to shoal, and this anti-predator behavior facilitates parasite transmission. To test for evidence of predator-driven differences in infection in natural populations, we collected 4715 guppies from 62 sites across Trinidad between 2003 and 2009 and screened them for ectosymbionts, including Gyrodactylus. A novel model-averaging analysis revealed that females were more likely to be infected with Gyrodactylus parasites than males, but only in populations with both high predation pressure and high infection prevalence. We propose that the difference in shoaling tendency between the sexes could explain the observed difference in infection prevalence between males and females in high-predation sites. The infection rate of juveniles did not vary with predation regime, probably because juveniles face constant predation pressure from conspecific adults and therefore tend to shoal in both high- and low-predation sites. This represents the first evidence for age- and sex-specific trait-mediated indirect effects of predators on the probability of infection in their prey.

Cryptic species complexes are common among parasites, which tend to have large populations and are subject to rapid evolution. Such complexes may arise through host-parasite co-evolution and/or host switching. For parasites that reproduce directly on their host, there might be increased opportunities for sympatric speciation, either by exploiting different hosts or different micro-habitats within the same host. The genus Gyrodactylus is a specious group of viviparous monogeneans. These ectoparasites transfer between teleosts during social contact and cause significant host mortality. Their impact on the guppy (Poecilia reticulata), an iconic evolutionary and ecological model species, is well established and yet the population genetics and phylogenetics of these parasites remains understudied. Using mtDNA sequencing of the host and its parasites, we provide evidence of cryptic speciation in Gyrodactylus bullatarudis, G. poeciliae and G. turnbulli. For the COII gene, genetic divergence of lineages within each parasite species ranged between 5.7 and 17.2%, which is typical of the divergence observed between described species in this genus. Different lineages of G. turnbulli and G. poeciliae appear geographically isolated, which could imply allopatric speciation. In addition, for G. poeciliae, co-evolution with a different host species cannot be discarded due to its host range. This parasite was originally described on P. caucana, but for the first time here it is also recorded on the guppy. The two cryptic lineages of G. bullatarudis showed considerable geographic overlap. G. bullatarudis has a known wide host range and it can also utilize a killifish (Anablepsoides hartii) as a temporary host. This killifish is capable of migrating overland and it could act as a transmission vector between otherwise isolated populations. Additional genetic markers are needed to confirm the presence of these cryptic Gyrodactylus species complexes, potentially leading to more in-depth genetic, ecological and evolutionary analyses on this multi-host-parasite system.

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non-stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI-, NI- and I-treated groups (94.5, 64 and 62.3 μg mg(-1), respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re-infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3-75%, higher than in the non-exposed control (40.6% mortality)., (© 2014 John Wiley & Sons Ltd.)

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.)

The unprecedented polymorphism in the major histocompatibility complex (MHC) genes is thought to be maintained by balancing selection from parasites. However, do parasites also drive divergence at MHC loci between host populations, or do the effects of balancing selection maintain similarities among populations? We examined MHC variation in populations of the livebearing fish Poecilia mexicana and characterized their parasite communities. Poecilia mexicana populations in the Cueva del Azufre system are locally adapted to darkness and the presence of toxic hydrogen sulphide, representing highly divergent ecotypes or incipient species. Parasite communities differed significantly across populations, and populations with higher parasite loads had higher levels of diversity at class II MHC genes. However, despite different parasite communities, marked divergence in adaptive traits and in neutral genetic markers, we found MHC alleles to be remarkably similar among host populations. Our findings indicate that balancing selection from parasites maintains immunogenetic diversity of hosts, but this process does not promote MHC divergence in this system. On the contrary, we suggest that balancing selection on immunogenetic loci may outweigh divergent selection causing divergence, thereby hindering host divergence and speciation. Our findings support the hypothesis that balancing selection maintains MHC similarities among lineages during and after speciation (trans-species evolution)., (© 2014 The Authors. Journal of Evolutionary Biology © 2014 European Society For Evolutionary Biology.)

Riverine fish are subjected to heterogeneous flow velocities and turbulence and may use this to their advantage by selecting regions that balance energy expenditure for station holding while maximizing energy gain through feeding opportunities. This study investigated microhabitat selection by guppies Poecilia reticulata in terms of flow characteristics generated by hemisphere boulders in an open channel flume. Velocity and turbulence influenced the variation in swimming behaviour with respect to size, sex and parasite intensity. With increasing body length, fish swam further and more frequently between boulder regions. Larger guppies spent more time in the areas of high-velocity and low-turbulence regions beside the boulders, whereas smaller guppies frequented the low-velocity and high-turbulence regions directly behind the boulders. Male guppies selected the regions of low velocity, indicating possible reduced swimming ability owing to hydrodynamic drag imposed by their fins. With increasing Gyrodactylus turnbulli burden, fish spent more time in regions with moderate velocity and lowest turbulent kinetic energy which were the most spatially and temporally homogeneous in terms of velocity and turbulence. These findings highlight the importance of heterogeneous flow conditions in river channel design owing to the behavioural variability within a species in response to velocity and turbulence.

A reduction in the strength of selection is expected to cause the evolution of reduced trait expression. Elimination of a parasite should thus cause the evolution of reduced resistance to that parasite. To test this prediction in nature, we studied the fourth- and eighth-generation descendants of guppies (Poecilia reticulata) introduced into four natural streams following experimental elimination of a common and deleterious parasite (Gyrodactylus spp.). After two generations of laboratory rearing to control for plasticity and maternal effects, we infected individual fish to assess their resistance to the parasite. Contrary to theoretical expectations, the introduced guppy populations had rapidly and repeatably evolved increased resistance to the now-absent parasite. This evolution was not owing to a resistance-tolerance trade-off, nor to differences in productivity among the sites. Instead, a leading candidate hypothesis is that the rapid life-history evolution typical in such introductions pleiotropically increases parasite resistance. Our study adds a new dimension to the growing evidence for contemporary evolution in the wild, and also points to the need for a re-consideration of simple expectations from host-parasite theory. In particular, our results highlight the need for increased consideration of multiple sources of selection and pleiotropy when studying evolution in natural contexts.

Parasites typically have low reproductive fitness on paratenic hosts. Such hosts offer other significant inclusive fitness benefits to parasites, however, such as increased mobility and migration potential. The parasite fauna of the guppy (Poecilia reticulata) is dominated by the directly transmitted ectoparasites Gyrodactylus bullatarudis and Gyrodactylus turnbulli. In the wild, close predatory and competitive interactions occur between the guppy and the killifish Rivulus hartii. Previous observations suggest that these fish can share gyrodactylids, so we tested experimentally whether these parasites can use R. hartii as an alternative host. In aquaria, G. bullatarudis was the only species able to transmit from prey to predator. Both parasite species transferred equally well to prey when the predator was experimentally infected. However, in semi-natural conditions, G. bullatarudis transmitted more successfully to the prey fish. Importantly, G. bullatarudis also survived significantly longer on R. hartii out of water. As R. hartii can migrate overland between isolated guppy populations, G. bullatarudis may have an enhanced ability to disperse and colonize new host populations, consistent with its wider distribution in the wild. To our knowledge, this is the first empirical study demonstrating a predator acting as a paratenic host for the parasites of its prey.

Studies of phenotypic variation in nature often consider only a single potential selective agent. In such cases, it remains an open question as to whether variation attributed to that single measured agent might be influenced by some other unmeasured agent. Previous research has shown that phenotypic variation in the Trinidadian guppy (Poecilia reticulata) is strongly influenced by predation regime, and we here ask whether parasitism might represent an additional important selective agent shaping this variation. We performed a field survey of 26 natural guppy populations of known predation regime in northern Trinidad. We quantified levels of parasitism of guppies by the monogenean ecotoparasite, Gyrodactylus, and examined whether this parasite was associated with guppy body size or male colour. Spatial variation in Gyrodactylus parasitism was consistent between years, and parasite prevalence was generally, but not always, higher at high-predation sites than at low-predation sites. Consistent with previous work, predation regime was related to guppy size and some aspects of male colour, whereas parasitism showed few and only minor associations with the same traits. Moreover, a consideration of parasitism did not alter any interpretations regarding associations between guppy traits and predation regimes. These results suggest that parasitism, at least as quantified in the present study, does not play a major role in shaping variation in guppy body size or colour. Nevertheless, considerable variation in these traits, even within a predation regime, suggests the likely importance of other selective agents beyond just predation regime.

Understanding disease transmission is important to species management and human health. Host body condition, nutrition and disease susceptibility interact in a complex manner, and while the individual effects of these variables are well known, our understanding of how they interact and translate to population dynamics is limited. Our objective was to determine whether host relative body condition influences epidemic dynamics, and how this relationship is affected by food availability. Poecilia reticulata (guppies) of roughly similar size were selected and assembled randomly into populations of 10 guppies assigned to 3 different food availability treatments, and the relative condition index (Kn) of each fish was calculated. We infected 1 individual per group ('source' fish) with Gyrodactyus turnbulli and counted parasites on each fish every other day for 10 days. Epidemic parameters for each population were analysed using generalized linear models. High host Kn-particularly that of the 'source' fish-exerted a positive effect on incidence, peak parasite burden, and the degree of parasite aggregation. Low food availability increased the strength of the associations with peak burden and aggregation. Our findings suggest that host Kn and food availability interact to influence epidemic dynamics, and that the condition of the individual that brings the parasite into the host population has a profound impact on the spread of infection.

Predation and parasitism are two of the most important sources of mortality in nature. By forming groups, individuals can gain protection against predators but may increase their risk of being infected with contagious parasites. Animals might resolve this conflict by forming mixed-species groups thereby reducing the costs associated with parasites through a relative decrease in available hosts. We tested this hypothesis in a system with two closely related poeciliid fishes (Poecilia reticulata and Poecilia picta) and their host-specific monogenean ectoparasites (Gyrodactylus spp.) in Trinidad. Fish from three different rivers were sampled from single and mixed-species groups, measured and scanned for Gyrodactylus. The presence and abundance of Gyrodactylus were lower when fish of both species were part of mixed-species groups relative to single-species groups. This is consistent with the hypothesis that mixed-species groups provide a level of protection against contagious parasites. We discuss the importance of potentially confounding factors such as salinity and individual fish size.

In a previous study we found that female guppies shoaled more than males and that there was greater transmission of the ectoparasite Gyrodactylus turnbulli between females. Here, to test for a possible sex bias in parasite transmission, we conducted a similar experiment on single sex shoals of male and female guppies, observing host behaviour before and after the introduction of an infected shoal mate. The initial parasite burden was considerably lower in the present experiment (30 worms versus >100 worms previously) and we used a different stock of ornamental guppies (Green Cobra variety versus a Tuxedo hybrid previously). Contrary to our previous finding, males aggregated significantly more than females. Males performed 'sigmoid' displays towards each other, a courtship behaviour that is more generally directed towards females. Due to the high rate of male-male interactions, parasite transmission was 10 times higher between males than between females. Furthermore, shoaling intensity was highest for the most parasitised fish indicating that these infected fish were not avoided by non-parasitised conspecifics. These studies show that certain social behaviours including shoaling and courtship displays, appear to facilitate the transmission of gyrodactylid parasites., (Copyright © 2012 Elsevier Inc. All rights reserved.)