Evolutionary Ecology Research, 2002, 4: 219226

Temporal and spatial distributions of parasites and

Department of Biology, Indiana University, Bloomington, IN 47405-3700, USA and

ABSTRACT
The Red Queen hypothesis predicts that sexual reproduction should be favoured in geographic
locations where the risk of infection is high. We surveyed 20 lakes on the South Island of
New Zealand to determine whether an association exists between the presence of individuals
sterilized by trematode larvae and the presence of males in a common freshwater snail
(Potamopyrgus antipodarum). The snail is unique in that populations are often mixtures of
triploid asexual females that reproduce by apomictic parthenogenesis and diploid sexual
individuals that reproduce by cross-fertilization. We compared the results to a similar study
conducted over 10 years ago. The results showed a highly signicant correlation between the
frequency of males in the past and present data. In addition, the results showed a strong
signicant correlation between the prevalence of trematodes in past and present data sets,
indicating that any selection imposed by the parasites has remained reasonably constant.
Finally, the frequency of males averaged over both data sets was signicantly and positively
correlated with the mean frequency of infected individuals. These results suggest that infection
levels and male frequency may be relatively stable and that parasites may be a factor in selecting
for sexual reproduction.
Keywords: Microphallus, New Zealand, Potamopyrgus antipodarum, Red Queen hypothesis,
sexual reproduction.

INTRODUCTION
The presence of sexual reproduction remains an unsolved problem in evolutionary biology
(Williams, 1975; Maynard Smith, 1978; Bell, 1982). The problem is especially clear in
populations where sexual and asexual females co-exist and sexual females invest equally in
male and female ospring. All else being equal, the asexuals should eliminate the sexuals,
given the two-fold reproductive advantage of asexual reproduction (Maynard Smith, 1978).
One possible solution, known as the Red Queen hypothesis, is that parasites evolve to infect
the most common host genotypes; as such, they select against any asexual form of reproduction once the clone has become common (Jaenike, 1978, Bremermann, 1980; Hamilton,
1980, 1982; Lloyd, 1980; Hamilton et al., 1990). The hypothesis has proven dicult to test
* Author to whom all correspondence should be addressed. e-mail: [email protected]
Consult the copyright statement on the inside front cover for non-commercial copying policies.
2002 Curtis M. Lively

220

Lively and Jokela

directly, but it makes several falsiable predictions. One key prediction is that, if wholly
parthenogenetic (or self-fertilizing) populations exist, they should be found in locations that
are relatively free of co-evolving parasites (Bell, 1982).
This prediction has support from a variety of studies (Glesener and Tilman, 1978; Bell,
1982; Lively, 1987, 1992; Schrag et al., 1994). In one of these studies, sexual reproduction in
a freshwater snail (Potamopyrgus antipodarum) was found to be positively and signicantly
correlated with prevalence of infection by larval trematodes (Lively, 1992). One weakness
of this study, however, was that the data were collected in a short window of time, so it is
unknown whether the pattern is temporally robust. Here we repeated the original survey by
sampling more intensively a subset of the original populations (n = 20). Our main aims
were: (1) to determine whether the frequency of sexual hosts or the prevalence of infection
had changed over the 1015 years between surveys; and (2) to determine whether the
previously observed correlation between sex and infection was repeatable.
NATURAL HISTORY OF THE HOST AND PARASITE
We focus on the most common trematode that infects the snail. This species is presently
undescribed, but is probably a new species in the genus Microphallus (S. Deblock, personal
communication); we refer to this species here and elsewhere as Microphallus sp. Previous
data show that Microphallus sp. infects P. antipodarum throughout New Zealand and that
there is considerable gene ow among geographically separated lake populations of the
worm (Dybdahl and Lively, 1996). Hence, the dierent populations of Microphallus do not
appear to represent dierent species, although they show strong adaptation to local host
populations (Lively, 1989).
The worm has a two-host life cycle. Snails are infected by ingesting eggs. A successful
infection yields several hundred asexually produced larvae, which encyst in the host snail.
Both sexes of this gonochoric, Prosobranch snail are sterilized during the course of the
development of these larval cysts. The cysts (called metacercariae) hatch when ingested by
a nal, vertebrate host (known to include dabbling ducks and black stilts) to yield mature
hermaphroditic adults in about 24 h. The tiny worms (0.2 mm long) then produce eggs
by cross-fertilization (as inferred from genetic data; Dybdahl and Lively, 1996), which are
infective to P. antipodarum. The risk of infection is expected to vary among locations if the
abundance of the nal hosts also varies.
The host snail is a small (47 mm long) gastropod that is common in New Zealands
freshwater habitats below tree line. Both sexuals and asexuals brood eggs in the mantle until
they hatch and the young crawl out of the brood pouch. Asexual females are triploid and
reproduction is by apomixis (Dybdahl and Lively, 1995). Sexual females are diploid and
cross-fertilization is required for the production of young. Sexual and asexual females taken
from the same lake and habitat mature at the same size and they produce the same number
of ospring (Jokela et al., 1997a). The frequency of developmental defects is also the same.
Hence, there would appear to be a full two-fold cost of sexual reproduction.
METHODS
We selected lakes that were among the most intensively sampled between 1985 and 1988
(Table 1). These glacial lakes, which were all formed following the Pleistocene, are
distributed on both sides of the Southern Alps in the central part of New Zealands South

Infection of a freshwater snail by parasites

221

Table 1. Sampling locations

Lake

Early
mean (n)

Late
mean (n)

Dierence

Alexandrina
Catherine
Clearwater
Grasmere
Haupiri
Hawdon
Ianthe
Ida
Kanieri
Katrine
Mapourika
Marymere
Moeraki
Paringa
Poerua
Sarah
Selfe
Sumner
Taylor
Tennyson

1986 (10)
1988 (1)
1988 (2)
1985 (2)
1987 (5)
1987 (2)
1987 (6)
1987 (3)
1986 (4)
1987 (1)
1987 (11)
1987 (3)
1988 (2)
1987 (2)
1987 (4)
1985 (3)
1988 (1)
1987 (1)
1987 (1)
1988 (5)

2000 (20)
2001 (1)
2000 (3)
1998 (4)
1998 (4)
1998 (11)
2000 (2)
1997 (2)
1998 (9)
2001 (4)
2000 (4)
2000 (3)
2000 (3)
2000 (3)
2000 (1*)
1998 (6)
1997 (6)
2001 (2)
2001 (4)
1996 (6)

14
13
12
13
11
11
13
10
12
14
13
13
12
13
13
13
9
14
14
8

Note: Entries give the lake names, the weighted mean year for the early (198588) and late
(mostly 19972001) samples with sample sizes in parentheses. The dierence between means is
given in the last column. The average dierence in years between the weighted means was 12.2.
* 234 snails from a 400 m transect along the shore.

Island. For these lakes, the correlation between (log) male frequency and (log) Microphallus
prevalence was not statistically signicant in the original survey (r = 0.333; n = 20; P =
0.152), although the correlation was signicant in the larger sample (Lively, 1992).
We sampled the shallow-water (< 0.5 m) margins of the lakes by washing snails from small
rocks into kick nets, or by pushing the nets through the shallow-water vegetation. Most
lakes were sampled between 1998 and 2001, with the exception that one of the lakes was
sampled in 1995 and 1996 and two of the lakes in 1997. Since we knew from previous
samples that there can be considerable spatial variation for infection in shallow water
(Jokela and Lively, 1995a,b), we sampled from one to 20 (usually four) locations in each lake
(depending on lake size and access). We then transported the snails to a eld station in
Kaikoura, New Zealand, where the snails were dissected (n > 100 for each sample). During
dissection, we recorded the gender of each snail and whether it was infected by trematode
larvae. We used male frequency as our measure of the frequency of diploid sexual females,
as these variables are highly positively correlated (Fox et al., 1996). The samples were then
averaged to give a single datum for each lake; the data were log-transformed to meet the
assumptions of parametric tests (Sokal and Rohlf, 1981). We report two-tailed probability
values.

222

Lively and Jokela

RESULTS
The results showed a strong positive correlation between male frequency in the early and
late samples (r = 0.875; n = 20; P < 0.001). This nding indicates that lakes having high
levels of sexual reproduction in the 1980s still have high levels of sexual reproduction
(Fig. 1). Similarly, there was a strong positive correlation between the level of infection by
Microphallus between the two sampling periods (Fig. 2), indicating that lakes previously
having high levels of infection still have high levels of infection (r = 0.864; n = 20; P < 0.001).
Thus, it would appear that, at least over this time period of 1015 years, any selection that
may have been imposed by parasites has remained reasonably constant. Finally, we found
that the frequency of males and prevalence of Microphallus were positively and signicantly
correlated in the new survey (r = 0.452; n = 20; P = 0.046), which is consistent with the
results from the earlier survey.
Based on these results, we reasoned that our best measures of parasite prevalence and
male frequency would be the means of both sampling periods. Interestingly, these means
for male frequency give a bimodal distribution, which is apparent from inspection
of Fig. 1. One mode exists at very low male frequency. The mean ( standard error)
of the populations associated with this mode is only 2.11 0.43. This low-male group
is likely to represent all clonal populations, since some clones are known to produce
up to 6% males. The populations associated with the second mode exists at a much higher
male frequency (23.49 2.41). This high-male group probably represents populations
composed of both sexual and asexual individuals (mixed populations; see Dybdahl and
Lively, 1995).
We then compared the prevalences of infection between these two groups of populations.
The high-male group had a higher mean prevalence of infection by Microphallus
(7.49 2.05) than the low-male group (2.49 0.60), and the dierence between means

Fig. 1. Bivariate plot of P. antipodarum male frequency from samples collected in the mid-1980s
against male frequency for samples collected primarily between 1997 and 2001. The solid line shows
the line of no change; deviations from the line give the dierences between the early and late samples.
Note that the points fall into two groups: one group for which male frequency is less that 6% for both
samples and another group for which male frequency is greater than 15%. The former (low-male)
group is likely to represent all clonal populations, since some clones are known to produce up to 6%
males. The latter (high-male) group represents populations composed of sexual and asexual individuals (mixed populations).

Infection of a freshwater snail by parasites

223

Fig. 2. Bivariate plot of prevalence of infection by Microphallus from samples collected in the
mid-1980s against prevalence of infection for samples collected primarily between 1997 and 2001. The
solid line shows the line of no change; deviations from the line give the dierences between the early
and late samples. Open circles show asexual populations; lled circles show mixed populations.

(log-transformed data) was statistically signicant (F1,18 = 4.74; P = 0.043). The result can be
seen in Fig. 2, which shows the prevalence of infection in mixed and asexual populations
for both sampling periods. The same result was obtained for total prevalence of infection
for all trematode species. The congruence of results, however, is not surprising given that
total prevalence and Microphallus prevalence are highly correlated.
DISCUSSION
In previous studies, we have used the snail P. antipodarum to test the hypothesis for the
evolutionary maintenance of sexual reproduction. At present, our results provide no evidence in support of the lottery model (Lively, 1987), which is consistent with Bells (1982)
broad taxonomic survey of the literature and with Schrag and co-workers (1994) study
of the distribution of outcrossing potential in a hermaphroditic snail. Our previous results
are also consistent with Bells nding that sex is more common in stable habitats, which
is consistent with both the tangled bank model and the Red Queen model, assuming
parasites are also more common in stable habitats (Lively, 1987). However, our results
also show that parasites account for more of the variation in male frequency than habitat
per se, which favours the Red Queen model, but does not eliminate the tangled bank
hypothesis (Lively, 1987). Finally, we have found no support for the deterministic mutation
hypothesis (Kondrashov, 1988; Charlesworth, 1990; Howard, 1994), at least in terms of
how it could be used to explain the distribution of parthenogenesis in these snails (Lively
et al., 1998).
The results of the study are also consistent with expectation under the Red Queen
hypothesis. We found that the prevalence of infection by a digenetic trematode was correlated between samples taken over a 1015 year period (Fig. 2), which suggests that any
selection imposed by this worm is relatively constant over time. If this were not the case, it
would be dicult to see how the parasites could be responsible for the maintenance of sex in
the highly sexual populations. Similarly, we found that male frequency was highly correlated

224

Lively and Jokela

between the two sampling periods and that there was no overall tendency for males to have
decreased over time (Fig. 1).
In addition, we found that male frequency and parasite prevalence were positively
correlated in both the new data set and over both data sets combined. We nd the results
from the combined data set to be particularly interesting, as it should give the best estimates
of the variables, and because male frequency broke cleanly into two groups of populations:
low-male populations, which are probably composed of asexual individuals, and high-male
populations, which are at least partially sexual. Our nding that the high-male populations
also have a signicantly higher mean level of infection is consistent with the Red Queen
hypothesis.
However, the result is also consistent with alternative hypotheses. For example, sex
and parasites would be expected to covary if sexual individuals, which are diploid, were
more susceptible to parasites than asexual individuals, which are triploid. We previously
examined this idea in one lake where sexuals and asexuals co-exist, but found no indication
that diploid individuals are inherently more infected by parasites (Jokela et al., 1997b)
or that males are inherently more infected than females (Lively, 1987, 2001). Hence,
it is unlikely that the association between sexual reproduction and parasites in the wild
is a simple consequence of sexuals being more susceptible to infection. In addition,
we have found in laboratory experiments that parasites are adapted to infecting the
most common triploid clones in a population composed entirely of asexual individuals
(Dybdahl and Lively, 1998). Hence, even if ploidy has some eect on inherent infectivity,
it would not appear to be sucient to prevent parasite tracking of locally common
triploid genotypes. Similarly, using reciprocal cross-infection experiments, we have found
that parasites from a mostly diploid host population were more likely to infect snails
from this same population than a remote triploid population of hosts and vice versa (Lively
and Dybdahl, 2000). Thus, local co-evolution would appear more important that any
inherent dierence in susceptibility between diploids and triploids, even if such dierences
do exist.
Other alternatives to explain the observed association between sex and parasites will
surely exist. These might be protably sought in those lakes with high male frequencies
and low parasite loads during both sampling periods (Fig. 2). However, in our minds, the
existence of such alternatives would not undermine the need for biogeographic data. We
feel that solid data on the distribution of asexual populations has been, and will continue to
be, important to the evaluation of the dierent hypotheses for sex (e.g. Glesener and
Tilman, 1978; Bell, 1982; Little and Ebert, 1999; Michiels et al., 2001; see Kondrashov,
1993, for an alternative view). Our approach has been to combine eld surveys with
laboratory experiments. Based on our results to date, we cannot eliminate the Red Queen
hypothesis.
In closing, we should point out an important subtlety. The Red Queen hypothesis
predicts that sexual reproduction should be found where the risk of exposure to virulent
parasites is high, but we have measured prevalence of infection instead of risk. It is reasonable to suspect that risk of exposure was underestimated. This may be especially true if,
in fact, the Red Queen hypothesis is correct, because greater levels of sex reduce the levels
of infection for any given value of risk (Lively, 2001). Thus, our measures may be disproportionately conservative for sexual populations. It would be interesting to discover
whether the association between sex and parasites reported here would become stronger if
risk of parasite exposure could be directly determined.

Infection of a freshwater snail by parasites

225

ACKNOWLEDGEMENTS
This paper was drawn from a symposium talk in memory of W.D. Hamilton at Kyushu University. We
wish to thank the organizers (Tet Yahara and Akira Sasaki) and participants for helpful comments
and discussion. We also thank two anonymous reviewers for helpful suggestions. Data collection was
supported by grants from the US National Science Foundation (grant DEB 9904840 to C.M.L.) and
the Swiss National Science Foundation (grant 36-59242.99 to J.J.).

REFERENCES
Bell, G. 1982. The Masterpiece of Nature: The Evolution and Genetics of Sexuality. Berkeley, CA:
University of California Press.
Bremermann, H.J. 1980. Sex and polymorphism as strategies in hostpathogen interactions. J. Theor.
Biol., 87: 671702.
Charlesworth, B. 1990. Mutationselection balance and the evolutionary advantage of sex and
recombination. Genet. Res. (Camb.), 55: 199221.
Dybdahl, M.F. and Lively, C.M. 1995. Diverse, endemic and polyphyletic clones in mixed
populations of a freshwater snail (Potamopyrgus antipodarum). J. Evol. Biol., 8: 385389.
Dybdahl, M.F. and Lively, C.M. 1996. The geography of coevolution: comparative population
structures for a snail and its trematode parasite. Evolution, 50: 22642275.
Dybdahl, M.F. and Lively, C.M. 1998. Hostparasite coevolution: evidence for rare advantage and
time-lagged selection in a natural population. Evolution, 52: 10571066.
Fox, J.A., Dybdahl, M.F., Jokela, J. and Lively, C.M. 1996. Genetic structure of coexisting sexual
and clonal subpopulations in a freshwater snail (Potamopyrgus antipodarum). Evolution,
50: 15411548.
Glesener, R.R. and Tilman, D. 1978. Sexuality and the components of environmental uncertainty:
clues from geographical parthenogenesis in terrestrial animals. Am. Nat., 112: 659673.
Hamilton, W.D. 1980. Sex versus non-sex versus parasite. Oikos, 35: 282290.
Hamilton, W.D. 1982. Pathogens as causes of genetic diversity in their host populations. In
Population Biology of Infectious Diseases (R.M. Anderson and R.M. May, eds), pp. 269296.
New York: Springer.
Hamilton, W.D., Axelrod, R. and Tanese, R. 1990. Sexual reproduction as an adaptation to resist
parasites (a review). Proc. Natl. Acad. Sci. USA, 87: 35663573.
Howard, R.S. 1994. Selection against deleterious mutations and the maintenance of biparental sex.
Theor. Pop. Biol., 45: 313323.
Jaenike, J. 1978. An hypothesis to account for the maintenance of sex within populations. Evol.
Theory, 3: 191194.
Jokela, J. and Lively, C.M. 1995a. Spatial variation for infection by digenetic trematodes in a
population of freshwater snails (Potamopyrgus antipodarum). Oecologia, 103: 509517.
Jokela, J. and Lively, C.M. 1995b. Parasites, sex, and early reproduction in a mixed population of
freshwater snails. Evolution, 49: 12681271.
Jokela, J., Lively, C.M., Dybdahl, M.F. and Fox, J.A. 1997a. Evidence for a cost of sex in the
freshwater snail Potamopyrgus antipodarum. Ecology, 78: 452460.
Jokela, J., Lively, C.M., Fox, J.A. and Dybdahl, M.F. 1997b. Flat reaction norms and frozen
phenotypic variation in clonal snails (Potamopyrgus antipodarum). Evolution, 51: 11201129.
Kondrashov, A.S. 1988. Deleterious mutations and the evolution of sexual reproduction. Nature,
336: 435440.
Kondrashov, A.S. 1993. Classication of hypotheses on the advantage of amphimixis. J. Hered., 84:
372387.
Little, T.J. and Ebert, D. 1999. Associations between parasitism and host genotype in natural
populations of Daphnia (Crustacea: Cladocera). J. Anim. Ecol., 68: 134149.

226

Lively and Jokela

Lively, C.M. 1987. Evidence from a New Zealand snail for the maintenance of sex by parasitism.
Nature, 328: 519521.
Lively, C.M. 1989. Adaptation by a parasitic trematode to local populations of its snail host.
Evolution, 43: 16631671.
Lively, C.M. 1992. Parthenogenesis in a freshwater snail: reproductive assurance versus parasitic
release. Evolution, 46: 907913.
Lively, C.M. 2001. Trematode infection and the distribution and dynamics of parthenogenetic snail
populations. Parasitology, 123: S19S26.
Lively, C.M. and Dybdahl, M.F. 2000. Parasite adaptation to locally common host genotypes.
Nature, 405: 679681.
Lively, C.M., Lyons, E.J., Peters, A.D. and Jokela, J. 1998. Environmental stress and the maintenance
of sex in a freshwater snail. Evolution, 52: 14821486.
Lloyd, D.G. 1980. Benets and handicaps of sexual reproduction. Evol. Biol., 13: 69111.
Maynard Smith, J. 1978. The Evolution of Sex. Cambridge: Cambridge University Press.
Michiels, N.K., Beukeboom, L.W., Pongraz, N. and Zeitlinger, J. 2001. Parthenogenetic atworms
have more symbionts than their coexisting, sexual conspecics but does this support the
Red Queen? J. Evol. Biol., 14: 110119.
Schrag, S.J., Mooers, A.O., Ndifon, G.T. and Read, A.F. 1994. Ecological correlates of male
outcrossing ability in a simultaneous hermaphrodite snail. Am. Nat., 143: 636655.
Sokal, R.R. and Rohlf, F.J. 1981. Biometry, 2nd edn. New York: W.H. Freeman.
Williams, G.C. 1975. Sex and Evolution. Princeton, NJ: Princeton University Press.