Presidential address: rediscovering parasites using molecular tools--towards revising the taxonomy of Echinococcus, Giardia and Cryptosporidium

Molecular biology has provided parasitologists with a fantastic variety of techniques that have had a major impact on research into parasites and parasitism. Molecular tools have revealed the extent and nature of genetic diversity in parasites and this information has made a significant contribution to studies on the population genetics and evolutionary biology of parasites. Similarly, epidemiology has benefited enormously from the application of molecular tools in terms of studying parasite life cycles and transmission, and in the development of specific and sensitive methods for diagnosis and surveillance. However, the theme I wish to develop in this paper is concerned with the contribution molecular tools have made to parasite taxonomy and systematics, and in particular, the fact that in many cases molecular tools are validating the proposals made many years ago by taxonomists and biologists which were discounted or not fully accepted at the time. To do this I have chosen four examples (Echinococcus, Entamoeba, Giardia, Cryptosporidium) where recent research involving molecular characterisation has confirmed observations made many years ago and has resulted in a need to revise the taxonomy of different groups of parasites.     

The adaptive significance of inquiline parasite workers

Social parasites exploit the socially managed resources of their host's society. Inquiline social parasites are dependent on their host throughout their life cycle, and so many of the traits inherited from their free-living ancestor are removed by natural selection. One trait that is commonly lost is the worker caste, the functions of which are adequately fulfilled by host workers. The few inquiline parasites that have retained a worker caste are thought to be at a transitional stage in the evolution of social parasitism, and their worker castes are considered vestigial and non-adaptive. However, this idea has not been tested. Furthermore, whether inquiline workers have an adaptive role outside the usual worker repertoire of foraging, brood care and colony maintenance has not been examined. In this paper, we present data that suggest that workers of the inquiline ant Acromyrmex insinuator play a vital role in ensuring the parasite's fitness. We show that the presence of these parasite workers has a positive effect on the production of parasite sexuals and a negative effect on the production of host sexuals. This suggests that inquiline workers play a vital role in suppressing host queen reproduction, thus promoting the rearing of parasite sexuals. To our knowledge, these are the first experiments on inquiline workers and the first to provide evidence that inquiline workers have an adaptive role.     

Ecology of helminth communities in tropical Australian amphibians.

Less than 50% of Australian amphibians have been recorded as hosts for helminth parasites. Despite developments in parasite community ecology in amphibians elsewhere, Australia lags behind with only two publications on this subject. Reasons advanced for this are that much of the collecting and taxonomic studies were conducted earlier this century before more recent discoveries of host genera and species as well as species complexes in the amphibian fauna. Consequently, there is a need for re-collection of hosts and parasites, and taxonomic revision of the parasites. In addition, as shown in this study, the parasite fauna in Australian amphibians is depauperate. Composition of the parasite fauna was largely dependent on the ecological associations of the host animal species. Frogs were infected with few helminth species and these occurred at low intensity, indicating, as in Europe and North America, that a depauperate fauna is also characteristic of amphibians in tropical regions.     

Patterns of host specificity and transmission among parasites of wild primates

Multihost parasites have been implicated in the emergence of new diseases in humans and wildlife, yet little is known about factors that influence the host range of parasites in natural populations. We used a comprehensive data set of 415 micro- and macroparasites reported from 119 wild primate hosts to investigate broad patterns of host specificity. The majority (68%) of primate parasites were reported to infect multiple host species, including animals from multiple families or orders. This pattern corresponds to previous studies of parasites found in humans and domesticated animals. Within three parasite groups (viruses, protozoans and helminths), we examined parasite taxonomy and transmission strategy in relation to measures of host specificity. Relative to other parasite groups, helminths were associated with the greatest levels of host specificity, whereas most viruses were reported to infect hosts from multiple families or orders. Highly significant associations between the degree of host specificity and transmission strategy arose within each parasite group, but not always in the same direction, suggesting that unique constraints influence the host range of parasites within each taxonomic group. Finally characteristics of over 100 parasite species shared between wild primates and humans, including those recognised as emerging in humans, revealed that most of these shared parasites were reported from multiple host orders. Furthermore, nearly all viruses that were reported to infect both humans and non-human primates were classified as emerging in humans.     

Genetic and immunological characterization of the 14-3-3xi molecule from Schistosoma bovis

Currently available candidate vaccines against schistosomiasis elicit only partial protection. In addition, the type of immune response that could lead to the highest level of protection against schistosomes has not yet been described. Thus, efforts should be made in both the identification of novel proteins essential for the parasite cycle and in the modulation of immune responses against these novel candidates through the combined use of immunomodulatory molecules. Several parasites have 14-3-3 proteins, and these proteins are known to play a key role in parasite biology. In the present work, we report the isolation and characterization of a new 14-3-3 gene from Schistosoma bovis and offer new information regarding the genetic structure of the gene. In addition, we have produced the corresponding recombinant protein. Finally, we describe the immune responses elicited by this protein when combined with 4 different immunomodulators in immunized mice.     

Species loss on spatial patterns and composition of zoonotic parasites

Species loss can result in the subsequent loss of affiliate species. Though largely ignored to date, these coextinctions can pose threats to human health by altering the composition, quantity and distribution of zoonotic parasites. We simulated host extinctions from more than 1300 host–parasite associations for 29 North American carnivores to investigate changes in parasite composition and species richness. We also explored the geography of zoonotic parasite richness under three carnivore composition scenarios and examined corresponding levels of human exposure. We found that changes in parasite assemblages differed among parasite groups. Because viruses tend to be generalists, the proportion of parasites that are viruses increased as more carnivores went extinct. Coextinction of carnivore parasites is unlikely to be common, given that few specialist parasites exploit hosts of conservation concern. However, local extirpations of widespread carnivore hosts can reduce overall zoonotic richness and shift distributions of parasite-rich areas. How biodiversity influences disease risks remains the subject of debate. Our results make clear that hosts vary in their contribution to human health risks. As a consequence, so too does the loss (or gain) of particular hosts. Anticipating changes in host composition in future environments may help inform parasite conservation and disease mitigation efforts.     

Partitioning the aggregation of parasites on hosts into intrinsic and extrinsic components via an extended Poisson-gamma mixture model

It is well known that parasites are often highly aggregated on their hosts such that relatively few individuals host the large majority of parasites. When the parasites are vectors of infectious disease, a key consequence of this aggregation can be increased disease transmission rates. The cause of this aggregation, however, is much less clear, especially for parasites such as arthropod vectors, which generally spend only a short time on their hosts. Regression-based analyses of ticks on various hosts have focused almost exclusively on identifying the intrinsic host characteristics associated with large burdens, but these efforts have had mixed results; most host traits examined have some small influence, but none are key. An alternative approach, the Poisson-gamma mixture distribution, has often been used to describe aggregated parasite distributions in a range of host/macroparasite systems, but lacks a clear mechanistic basis. Here, we extend this framework by linking it to a general model of parasite accumulation. Then, focusing on blacklegged ticks (Ixodes scapularis) on mice (Peromyscus leucopus), we fit the extended model to the best currently available larval tick burden datasets via hierarchical Bayesian methods, and use it to explore the relative contributions of intrinsic and extrinsic factors on observed tick burdens. Our results suggest that simple bad luck-inhabiting a home range with high vector density-may play a much larger role in determining parasite burdens than is currently appreciated.     

Why do fish have so few roundworm (nematode) parasites?

Synopsis Although they are the oldest and most diverse members of the subphylum, the fishes have relatively few nematode parasites in comparison with other vertebrate classes. It is hypothesized that this paucity of parasite species has occurred because nematode parasites first evolved in terrestrial hosts and only a few lines of these parasites were able to transfer to fish after the appearance of heteroxeny (use of intermediate hosts) and paratenesis (use of transport hosts). The inability of nematodes to initiate parasitism in aquatic ecosystems restricted fish parasites mainly to forms first adapted to terrestrial vertebrates and at the same time deprived large groups of aquatic invertebrates such as the crustaceans, annelids and molluscs of a nematode parasite fauna.Invited editorial     

Molecular characterization of an ancient Hepatozoon species parasitizing the 'living fossil' marsupial 'Monito del Monte' Dromiciops gliroides from Chile

The Microbiotheriid Dromiciops gliroides , also known as 'Monito del Monte', is considered to be a threatened species and the only living representative of this group of South American marsupials. During the last few years, several blood samples from specimens of 'Monito del Monte' captured at Chiloé island in Chile have been investigated for blood parasites. Inspection of blood smears detected a Hepatozoon species infecting red blood cells. The sequences of DNA fragments corresponding to small subunit ribosomal RNA gene revealed two parasitic lineages belonging to Hepatozoon genus. These parasite lineages showed a basal position with respect to Hepatozoon species infecting rodents, reptiles, and amphibians but are phylogenetically distinct from Hepatozoon species infecting the order Carnivora. In addition, the Hepatozoon lineages infecting D. gliroides are also different from those infecting other micro-mammals living in sympatry, as well as from some that have been described to infect an Australian species of bandicoot. The potential vector of this parasite appears to be the host-specific tick Ixodes neuquenensis because the sequencing of a long amplicon determined the presence of one of the two lineages found in the marsupial.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 568–576.     

Vaccination against cysticercosis and hydatid disease

Infections with the larval stages of taeniid cestode parasites cause substantial human morbidity as well as economic losses in domestic livestock species. Despite ongoing efforts around the world, few countries have been able substantially to reduce or eradicate these infections through the use of anthelmintics and lifestyle changes. Vaccines offer an additional potential tool to assist with the control of parasite transmission. Here, Marshall Lightowlers and colleagues review the substantial progress that has been made towards developing practical vaccines against hydatid disease in sheep and cysticercosis in sheep and cattle. Recombinant antigens have been used to induce more than 90% protection against challenge infections. Such success in animals encourages investigation of the potential use of vaccines in humans to prevent hydatid disease arising from infection with Echinococcus granulosus and cysticercosis from infection with Taenia solium.     

Notes on some petrified cones of the Pinaceae from the Cretaceous

References are made to five species of Pinaeeous cones from the Cretaceous. The general morphology of the first three was described by Fliche in 1896. Detailed studies on their anatomy have now enabled revisions to be made in their taxonomy. Pinus argonnensis Fliche has been shown to be indistinguishable from Pityoslrobus soigniesiensis Alvin, described in 1960. Tsugiles rnagnus Fliche and Pinus praemonticola Fliche are specimens of a previously named species, Pityostrobus andraei (Coemans) Seward, and their names have been included in its synonymy. A note on Pityostrobus leckenbyi (Lindley and Hutton) Seward attempts to clarify a few points on its anatomy and taxonomy. Finally the discovery of an additional specimen of Pityoslrobus jacksonii Creber is recorded and some details of this specimen are given.     

Correlating early evolution of parasitic platyhelminths to Gondwana breakup

Investigating patterns and processes of parasite diversification over ancient geological periods should involve comparisons of host and parasite phylogenies in a biogeographic context. It has been shown previously that the geographical distribution of host-specific parasites of sarcopterygians was guided, from Palaeozoic to Cainozoic times, mostly by evolution and diversification of their freshwater hosts. Here, we propose phylogenies of neobatrachian frogs and their specific parasites (Platyhelminthes, Monogenea) to investigate coevolutionary processes and historical biogeography of polystomes and further discuss all the possible assumptions that may account for the early evolution of these parasites. Phylogenetic analyses of concatenated rRNA nuclear genes (18S and partial 28S) supplemented by cophylogenetic and biogeographic vicariance analyses reveal four main parasite lineages that can be ascribed to centers of diversity, namely Australia, India, Africa, and South America. In addition, the relationships among these biogeographical monophyletic groups, substantiated by molecular dating, reflect sequential origins during the breakup of Gondwana. The Australian polystome lineage may have been isolated during the first stages of the breakup, whereas the Indian lineage would have arisen after the complete separation of western and eastern Gondwanan components. Next, polystomes would have codiverged with hyloid sensu stricto and ranoid frog lineages before the completion of South American and African plate separation. Ultimately, they would have undergone an extensive diversification in South America when their ancestral host families diversified. Therefore, the presence of polystome parasites in specific anuran host clades and in discrete geographic areas reveals the importance of biogeographic vicariance in diversification processes and supports the occurrence and radiation of amphibians over ancient and recent geological periods.     

Worker genetic diversity and infection by Nosema apis in honey bee colonies.

The hypothesis that parasites and pathogens select for polyandry in eusocial Hymenoptera was tested, using the honey bee Apis mellifera and its microsporidian parasite Nosema apis. Five honey bee colonies with low and five with high worker genetic diversity were infected with N. apis spores. At 54-56 days after inoculation, parasite spores in the workers' midguts were counted to determine whether there was a greater variation of infection intensity (spore counts per worker) in high-diversity colonies than in low-diversity ones. In all colonies there were two discrete sets of workers, with few or many parasite spores. To compare the variations of infection intensity between two colony groups, coefficients of variation were calculated for all workers examined, and for the slightly infected and strongly infected workers. The percentages of slightly infected workers in the low- and high-diversity groups were also compared. None of the comparisons between low- and high-diversity colonies showed significant differences, therefore no relation was found between honey bee workers' genetic diversity and their infection with N. apis.     

Endocrine relationships between nematodes and their insect hosts--a review

This paper reviews the evidence for a possible endocrine basis for the remarkable synchrony often observed between the development of entomophilic nematodes and that of their hosts. Although some suggestive observations have been made by various workers, there is no rigorous evidence which points to such an association. In those few cases which have been rigorously examined, there is some evidence that the synchrony is not based upon endocrine signals passing between host and parasite. The possibility that the parasite may manipulate the endocrinology of the insect in order to alter the metabolism and physiology of the host to favour the parasite has received less attention, but is nevertheless thought to constitute a promising areas of research.     

Oh sister,where art thou? Spatial population structure and the evolution of an altruistic defence trait

The evolution of parasite virulence and host defences is affected by population structure. This effect has been confirmed in studies focusing on large spatial scales, whereas the importance of local structure is not well understood. Slavemaking ants are social parasites that exploit workers of another species to rear their offspring. Enslaved workers of the host species Temnothorax longispinosus have been found to exhibit an effective post‐enslavement defence behaviour: enslaved workers were observed killing a large proportion of the parasites’ offspring. As enslaved workers do not reproduce, they gain no direct fitness benefit from this ‘rebellion’ behaviour. However, there may be an indirect benefit: neighbouring host nests that are related to ‘rebel’ nests can benefit from a reduced raiding pressure, as a result of the reduction in parasite nest size due to the enslaved workers’ killing behaviour. We use a simple mathematical model to examine whether the small‐scale population structure of the host species could explain the evolution of this potentially altruistic defence trait against slavemaking ants. We find that this is the case if enslaved host workers are related to nearby host nests. In a population genetic study, we confirm that enslaved workers are, indeed, more closely related to host nests within the raiding range of their resident slavemaker nest, than to host nests outside the raiding range. This small‐scale population structure seems to be a result of polydomy (e.g. the occupation of several nests in close proximity by a single colony) and could have enabled the evolution of ‘rebellion’ by kin selection.     

First report of three Kudoa species from eastern Australia: Kudoa thyrsites from mahi mahi (Coryphaena hippurus), Kudoa amamiensis and Kudoa minithyrsites n. sp. from sweeper (Pempheris ypsilychnus)

Fish species around the world are parasitized by myxozoans of the genus Kudoa, several of which infect and cause damage of commercial importance. In particular, Kudoa thyrsites and Kudoa amamiensis infect certain cultured fish species causing damage to muscle tissue, making the fish unmarketable. Kudoa thyrsites has a broad host and geographic range infecting over 35 different fish species worldwide, while K. amamiensis has only been reported from a few species in Japanese waters. Through morphological and molecular analyses we have confirmed the presence of both of these parasites in eastern Australian waters. In addition, a novel Kudoa species was identified, having stellate spores, with one polar capsule larger than the other three. The SSU rDNA sequence of this parasite was 1.5% different from K. thyrsites and is an outlier from K. thyrsites representatives in a phylogenetic analysis. Furthermore, the spores of this parasite are distinctly smaller than those of K. thyrsites, and thus it is described as Kudoa minithyrsites n. sp. Although the potential effects of K. minithyrsites n. sp. on its fish hosts are unknown, both K. thyrsites and K. amamiensis are associated with flesh quality problems in some cultured species and may be potential threats to an expanding aquaculture industry in Australia.     

LINKAGE BETWEEN NUCLEAR AND MITOCHONDRIAL DNA SEQUENCES IN AVIAN MALARIA PARASITES: MULTIPLE CASES OF CRYPTIC SPECIATION?

Analyses of mitochondrial cytochrome b diversity among avian blood parasites of the genera Haemoproteus and Plasmodium suggest that there might be as many lineages of parasites as there are species of birds. This is in sharp contrast to the approximately 175 parasite species described by traditional methods based on morphology using light microscopy. Until now it has not been clear to what extent parasite mitochondrial DNA lineage diversity reflects intra- or interspecific variation. We have sequenced part of a fast-evolving nuclear gene, dihydrofolate reductase-thymidylate synthase (DHFR-TS), and demonstrate that most of the parasite mitochondrial DNA lineages are associated with unique gene copies at this locus. Although these parasite lineages sometimes coexist in the same host individual, they apparently do not recombine and could therefore be considered as functionally distinct evolutionary entities, with independent evolutionary potential. Studies examining parasite virulence and host immune systems must consider this remarkable diversity of avian malaria parasites.