On the arrival of fasciolosis in the Americas.

Fasciola hepatica is a worldwide emerging and re-emerging parasite heavily affecting several regions in South America. Some lymnaeid snail species of American origin are among the major hosts of F. hepatica worldwide. Recent paleoparasitological findings detected its DNA in a 2300-year-old sample in Patagonia, countering the common hypothesis of the recent arrival of F. hepatica in the Americas during European colonization. Thus, the theory of an initial introduction in the 1500s can no longer be sustained. This article discusses how it was possible for F. hepatica to reach and spread in the Americas in relation to the availability and compatibility of hosts through natural and incidental introductions. Our study will serve to better understand the ongoing Neotropical scenario of fasciolosis.

An outbreak of canine schistosomiasis in Utah: Acquisition of a new snail host (Galba humilis) by Heterobilharzia americana, a pathogenic parasite on the move.

Parasites with complex life cycles engaging multiple host species living among different environments well-exemplify the value of a cross-cutting One Health approach to understanding fundamental concerns like disease emergence or spread. Here we provide new information regarding a pathogenic schistosome trematode parasite of both wild and domestic mammals that has recently expanded its known range from mesic/wet environments of the southeastern United States to the arid southwest. In 2018, 12 dogs living near a man-made pond in Moab, Utah, were found positive for Heterobilharzia americana, the most westerly report of this endemic North American schistosome, and the first from Utah. Raccoon scats collected near the pond were positive for H. americana eggs, and snails living near the pond's water line identified as Galba humilis shed H. americana cercariae, the first indication of natural infections in this widespread North American snail species. The susceptibility of G. humilis to H. americana was confirmed experimentally. Our studies support the existence of two variants of H. americana and emphasize the need for further investigations of lymnaeids and their compatibility with H. americana, to better define the future potential for its spread. Capture of a new species of intermediate host vector snail and construction of man-made habitats suitable for this snail have created the potential for a much more widespread animal health problem, especially for dogs and horses. H. americana will prove difficult to control because of the role of raccoons in maintaining transmission and the amphibious habits of the snail hosts of this pathogenic schistosome.

Genetic diversity and relationships of the liver fluke Fasciola hepatica (Trematoda) with native and introduced definitive and intermediate hosts.

Fasciolosis is a worldwide spread parasitosis mainly caused by the trematode Fasciola hepatica. This disease is particularly important for public health in tropical regions, but it can also affect the economies of many developed countries due to large infections in domestic animals. Although several studies have tried to understand the transmission by studying the prevalence of different host species, only a few have used population genetic approaches to understand the links between domestic and wildlife infections. Here, we present the results of such genetic approach combined with classical parasitological data (prevalence and intensity) by studying domestic and wild definitive hosts from Camargue (southern France) where fasciolosis is considered as a problem. We found 60% of domestic hosts (cattle) infected with F. hepatica but lower values in wild hosts (nutria, 19%; wild boars, 4.5%). We explored nine variable microsatellite loci for 1,148 adult flukes recovered from four different populations (non-treated cattle, treated cattle, nutria and wild boars). Populations from the four groups differed, though we found a number of migrants particularly non-treated cattle and nutria. Overall, we detected 729 different multilocus genotypes (from 783 completely genotyped individuals) and only 46 genotypes repeated across samples. Finally, we experimentally infected native and introduced intermediate snail hosts to explore their compatibility with F. hepatica and assess the risks of fasciolosis expansion in the region. The introduced species Galba truncatula and Pseudosuccinea columella attained the higher values of overall compatibility in relation to the European species. However, concerning the origin, sympatric combinations of G. truncatula were more compatible (higher prevalence, intensity and survival) than the allopatric tested. According to our results, we should note that the assessment of epidemiological risks cannot be limited to a single host-parasite system, but should focus on understanding the diversity of hosts in the heterogeneous environment through space and time.

Systematics and geographical distribution of Galba species, a group of cryptic and worldwide freshwater snails.

Cryptic species can present a significant challenge to the application of systematic and biogeographic principles, especially if they are invasive or transmit parasites or pathogens. Detecting cryptic species requires a pluralistic approach in which molecular markers facilitate the detection of coherent taxonomic units that can then be analyzed using various traits (e.g., internal morphology) and crosses. In asexual or self-fertilizing species, the latter criteria are of limited use. We studied a group of cryptic freshwater snails (genus Galba) from the family Lymnaeidae that have invaded almost all continents, reproducing mainly by self-fertilization and transmitting liver flukes to humans and livestock. We aim to clarify the systematics, distribution, and phylogeny of these species with an integrative approach that includes morphology, molecular markers, wide-scale sampling across America, and data retrieved from GenBank (to include Old World samples). Our phylogenetic analysis suggests that the genus Galba originated ca. 22 Myr ago and today comprises six species or species complexes. Four of them show an elongated-shell cryptic phenotype and exhibit wide variation in their genetic diversity, geographic distribution, and invasiveness. The remaining two species have more geographically restricted distributions and exhibit a globose-shell cryptic phenotype, most likely phylogenetically derived from the elongated one. We emphasize that no Galba species should be identified without molecular markers. We also discuss several hypotheses that can explain the origin of cryptic species in Galba, such as convergence and morphological stasis.

Prevalence of Fasciola hepatica infection in Galba cousini and Galba schirazensis from an Andean region of Ecuador.

Some Lymnaeid snails are intermediate hosts of the liver fluke Fasciola hepatica, the causal agent of fasciolosis, a zoonotic parasitic disease. Human and livestock fasciolosis has been reported in a highland community located in the Chimborazo Province of the Ecuadorian Andes. However, no previous study has been carried out to identify which snail species act as intermediate host/s of F. hepatica. This study first aimed to identify the intermediate snail species and secondly to determine the prevalence of natural infection with F. hepatica in 230 lymnaeid snails sampled from irrigation and drainage canals in this area. The first objective entailed observations of shell morphology and internal organs as well as sequencing of the cytochrome oxidase subunit 1 (COI) gene. For the second objective, we used classic parasitological methods (observation of rediae and cercarial emission) and PCR amplification specie-specific to F. hepatica. COI haplotype networks were built to elucidate phylogeographic relationships between the snail populations from this highland community with other American and worldwide populations. We identified two lymnaeid Galba cousini and Galba schirazensis and found high infection rates of F. hepatica in G. cousini, but these differed according to the method used, with PCR showing a higher rate (61 ± 20%) compared to rediae observation (29 ± 17%). F. hepatica in G. schirazensis was identified only by DNA amplification. G. cousini populations were genetically structured by geographic distance whereas G. schirazensis populations showed very low genetic diversity. The higher abundance and infection rate of G. cousini compared to G. schirazensis suggests that the former is likely the specie responsible for F. hepatica transmission in this region.

Reviewing Fasciola hepatica transmission in the West Indies and novel perceptions from experimental infections of sympatric vs. allopatric snail/fluke combinations.

Fasciolosis is an important food-borne parasitic disease affecting over two million people worldwide with economic losses related to cattle production of up to US$ 3 billion annually. Despite the long known presence of Fasciola hepatica in the Caribbean islands its transmission is not well known. This study reviews historical and recent data on fasciolosis in the West Indies, revealing for the first time the outcomes of sympatric and allopatric fluke/snail interactions in the area by exploring the susceptibility of four lymnaeid species after exposure to F. hepatica isolates from Cuba, the Dominican Republic and France. Overall, Galba cubensis showed a mean prevalence of 71.8% and appears to be the most suitable intermediate host species irrespective of the isolate used. Sympatric combinations (snail and parasite from the same country) were generally more compatible (higher susceptibility, parasite intensity and snail survival post-exposure) and only the allopatric interaction of French G. truncatula/Cuban F. hepatica attained 100% prevalence and mean intensity over 33 rediae/snail. However, certain Dominican populations of Pseudosuccinea columella showed high parasite intensities (>30 rediae/snail) when infected with Cuban flukes, highlighting the potential risks of biological introductions. Overall, high compatibility in most sympatric combinations compared to low or moderate compatibility in allopatric ones, suggests the existence of local adaptation from a long sustained interaction that has led to high rates of transmission. Interestingly, attempts to infect G. schirazensis with sympatric and allopatric flukes failed and coupled with the lowest survival rates which supposes a low risk of fasciolosis transmission in areas where this is the only snail species. Although there are significant gaps in the actual status of fasciolosis transmission from several islands in the West Indies these results show a permanent risk. We conclude that fasciolosis transmission is high in areas where the local snail, G. cubensis, occurs, and will be even higher in the presence of the invasive P. columella.

A strong colonizer rules the trematode guild in an intertidal snail host.

We examined the extent to which supply-side, niche, and competition theories and concepts help explain a trematode community in which one species comprises 87% of the trematode individuals, and the remaining 15 species each have <3%. We collected and dissected the common and wide-ranging snail host Heleobia australis over four seasons from three distinct habitats from the intertidal area of the Bahía Blanca estuary, Argentina. Inside a snail, trematodes interact with each other with outcomes that depend on facilitation, competition, and preemption, suggesting that dominant species should be common. The abundant trematode species, Microphallus simillimus, is a weak competitor, but has life-history traits and strategies associated with higher colonization ability that could increase its probability of invading the host first, allowing it to preempt the rare species. Rather than segregate by habitat, trematode species aggregated in pans during the summer where dominant trematode species often excluded subordinate ones. Despite losses to competition, and a lack of niche partitioning, M. simillimus ruled this species-rich trematode guild through strong recruitment and (potentially) preemption. Therefore, extremely skewed species abundance distributions, like this one, can derive from extremely skewed colonization abilities.

Impact of Human Activities on Fasciolosis Transmission.

Fasciolosis is a worldwide disease caused by the liver fluke Fasciola spp. This food- and water-borne disease is a major public health and veterinary issue. It is currently (re)emerging in several regions mainly due to the rapid evolution of human activities. This article reviews the current knowledge of the impact of irrigation-system management, livestock management, and human diet and hygiene habits on the emergence of fasciolosis. We also identify the gaps in this knowledge and the possible solutions for limiting these impacts. Integrated control seems to be the most effective solution for controlling fasciolosis, because it enables monitoring, prevention, and rapid action in case of the (re)emergence of the disease.

A new multiplex PCR assay to distinguish among three cryptic Galba species, intermediate hosts of Fasciola hepatica.

A molecular tool described here allows in one step for specific discrimination among three cryptic freshwater snail species (genus Galba) involved in fasciolosis transmission, a worldwide infectious disease of humans and livestock. The multiplex PCR approach taken targets for each species a distinctive, known microsatellite locus which is amplified using specific primers designed to generate an amplicon of a distinctive size that can be readily separated from the amplicons of the other two species on an agarose gel. In this way, the three Galba species (G. cubensis, G. schirazensis, and G. truncatula) can be differentiated from one another, including even if DNA from all three were present in the same reaction. The accuracy of this new molecular tool was tested and validated by comparing multiplex PCR results with species identification based on sequences at mitochondrial and nuclear markers. This new method is accurate, inexpensive, simple, rapid, and can be adapted to handle large sample sizes. It will be helpful for monitoring invasion of Galba species and for developing strategies to limit the snail species involved in the emergence or re-emergence of fasciolosis.

First molecular identification of Ascocotyle (Phagicola) longa in its first intermediate host the mud snail Heleobia australis.

This is the first study that used species-specific DNA primers to confirm the presence of the heterophyid Ascocotyle (Phagicola) longa Ransom, 1920 in its first intermediate host. The larval stages (rediae and cercariae) of this parasite were morphologically and genetically identified in the gonad of the intertidal mud snail Heleobia australis (d'Orbigny, 1835) (Cochliopidae) in the Bahía Blanca estuary, Argentina. In addition, we asked whether the prevalence in H. australis varied between seasons. Mullets - the second intermediate host of this heterophyid - migrate in estuaries during the warmer seasons and it is expected that piscivorous birds and mammals - the definitive hosts - prey more intensively on this species at those times. Thus, the number of parasite eggs released into the tidal flat within their feces should be higher, thereby increasing the ingestion of the parasite by H. australis.We therefore expected a higher prevalence of A. (P.) longa in H. australis in the Bahía Blanca estuary during spring and summer than autumn and winter. We found that 16 out of 2,744 specimens of H. australis had been infected with A. (P.) longa (total prevalence of 0.58%). Nonetheless, the prevalence showed no significant variation between seasons. Hence, we discuss an alternative scenario where the lack of seasonal changes might be mostly related to the permanent residence of definitive hosts in the estuary and not to the seasonal recruitment of mullets. Finally, we highlight the need for more experimental and comparative approaches in order to understand the diagnosis and geographical distribution of this worldwide heterophyid.

Larval trematodes infecting the South-American intertidal mud snail Heleobia australis (Rissooidea: Cochliopidae).

Larval trematodes infecting the snail Heleobia australis (Cochliopidae) from the Bahía Blanca estuary, Argentina were surveyed for two years. A total of 7,504 snail specimens was dissected and the larval stages of 15 different trematodes were recovered and examined morphologically. These larvae included four species that had previously been reported from H. australis in Brazil, Uruguay, and Argentina: the heterophyid Ascocotyle (Phagicola) longa and the microphallids, Maritrema bonaerense, Maritrema orensense, and Microphallus simillimus. Three other species, the echinostomatid Stephanoprora uruguayensis, the microphallid Levinseniella cruzi, and the psilostomid Psilochasmus oxyurus are reported here for the first time as parasites of H. australis. Eight other trematodes found in H. australis are described and placed in the appropriate superfamilies, families or genera (Cryptogonimidae, Apocreadiidae, Aporocotylidae, Notocotylidae, Haploporidae, Renicolidae, Himasthla, and Renicola). The prevalence of the trematode taxa infecting H. australis in the Bahía Blanca estuary was low (<3%) with a single exception (M. simillimus; >20%). Microphallidae were the richest and the most prevalent family, probably because of the high abundance of crabs - the second-intermediate hosts of certain microphallid species - and the considerable diversity of gulls. Here we compare the parasite assemblage found in the H. australis from Bahía Blanca estuary with other parasite assemblages infecting Heleobia spp. and other rissooids from the rest of the world.

Maritrema orensense and Maritrema bonaerense (Digenea: Microphallidae): descriptions, life cycles, and comparative morphometric analyses.

We elucidate the life cycle of Maritrema orensense for the first time and experimentally confirm that of the sympatric Maritrema bonaerense. In Argentinean estuaries, both species parasitize the cochliopid snail Heleobia australis as first intermediate host, the grapsid crabs Neohelice granulata and Cyrtograpsus angulatus as second intermediate hosts, and gulls as definitive hosts. Here, we describe the daughter sporocysts and cercariae of M. orensense and redescribe these stages for M. bonaerense. Sporocysts of M. orensense are shorter, with fewer developed cercariae than M. bonaerense. The cercariae of M. orensense have longer, larger, and more undulating cephalic glands than M. bonaerense. We redescribe metacercariae and adults of both species and compare them with the previous descriptions. Intestinal ceca length, vitellaria shape and extension, and egg size are the most relevant characteristics in metacercariae and adults for differentiating the species. Hence, the detailed morphological description and comparative analyses of morphometrics obtained from natural and experimental infections permit clear differentiation of M. orensense and M. bonaerense at each life stage.