Ecological factors responsible for the geographic distribution of Rhabdias joaquinensis: where do lungworms infect anurans in nature?

The lungworm Rhabdias joaquinensis (Nematoda: Rhabdiasidae) is a common parasite of anurans in eastern Nebraska, yet absent from the same hosts in western Nebraska. This study investigated the ecology of the lungworm's free-living stages that reside in host feces and surrounding soils to establish the worm's free-living niche in eastern Nebraska. Using a comparative method, this study also investigated the absence of lungworms in western Nebraska's Sandhills. Soil composition, moisture, and temperature were experimentally varied in the laboratory to assess their effects on juvenile development and survival. Field mesocosm and host defecation experiments were used to determine where in nature lungworms survive and eventually infect frogs and toads and to discover if vegetation facilitates lungworm transmission to hosts. The results found loam soils were amenable to lungworm development, whereas soils with high clay or sand content produced few infective lungworms. Soil moisture <50 % did not support lungworm development. Infective juveniles successfully developed between 5 and 35 °C, albeit at different rates, whereas only a limited number of non-infective worms developed at 40 °C. Field studies found that shoreline environments supported lungworm development, and the majority of lungworm infections appear to occur within a zone of infection adjacent to shorelines in eastern Nebraska. The prevalence in vegetation mesocosms was 100 %, and a significantly higher mean abundance was found in toads from containers with vegetation than without. Overall, these experiments suggest that the spatial distribution of R. joaquinensis in Nebraska is constrained by the worm's ability to survive and reproduce in a soil matrix.

Macroparasite population dynamics among geographical localities and host life cycle stages: eugregarines in Ischnura verticalis.

Populations of several species of gregarine parasites within a single host species, the damselfly Ischnura verticalis , were examined over the course of 1 season at 4 geographic localities separated by a maximum distance of 9.7 km. Gregarines, having a life cycle with both exogenous and endogenous stages, are subject to a wide variety of selective pressures that may drive adaptation. Gregarine species showed some specificity for host life cycle stage, i.e., Steganorhynchus dunwoodyi and Hoplorhynchus acanthatholius were most prevalent in larval hosts while Steganorhynchus dunwoodyi, Actinocephalus carrilynnae, and Nubenocephalus nebraskensis were most prevalent in adult hosts. Species prevalence and abundance differed by geographic locality. Gregarine prevalence was significantly higher in adult female damselflies than in males at 2 localities; sex differences in prevalence were insignificant for larval damselflies at all 4 localities. In larval hosts, gregarine abundance was independent of age (size). The present study, therefore, shows that pond characteristics, host life cycle stage, and adult host sex are the main factors that influence the prevalence and abundance of gregarine populations.

Co-occurrence of Haematoloechus complexus and Rhabdias joaquinensis in the plains leopard frog from Nebraska.

Species of Haematoloechus and Rhabdias both are ubiquitous lung parasites of frogs, yet surprisingly little research has been conducted on the interactions between these worms. In a small Nebraska stream, 256 Rhabdias joaquinensis and 225 Haematoloechus complexus were found in the lungs of Lithobates (=Rana) blairi. Thirty-six of the 44 (82%) frogs were co-infected with H. complexus and R. joaquinensis whereas 5 (11%) frogs were infected with only 1 species and 3 (7%) frogs were uninfected. Tests for association between H. complexus and R. joaquinensis found a significant positive relationship between the 2 parasites. Significant positive relationships in intensity of infection were found when both lungs were combined but not when lungs were considered individually. Tests for association between H. complexus and R. joaquinensis found no relationship between the worms in the anterior and posterior portions of the lungs. These data suggest that H. complexus and R. joaquinensis do not competitively exclude each other from the lungs of L. blairi in southeastern Nebraska.

Host specificity of North American Rhabdias spp. (Nematoda: Rhabdiasidae): combining field data and experimental infections with a molecular phylogeny.

Lungworms of the cosmopolitan genus Rhabdias are among the most common parasites of amphibians and squamate reptiles. The present study used experimental infections, field studies, and a molecular phylogeny to determine the host specificity of 6 Rhabdias spp. that infect snakes and anurans from North America. The molecular phylogeny suggests Rhabdias ranae from Nebraska and Mississippi may represent separate, cryptic species. In addition, the phylogeny strongly supports separate clades for anuran and snake lungworms. Field studies and experimental infections indicate that snake lungworms are generalist snake parasites; however, laboratory experiments also suggest that lizards can be infected under some environmental conditions. Lungworms from anurans were found not to infect salamanders or reptiles, in nature or in the laboratory; anuran lungworm species ranged from strict host specificity, e.g., R. ranae from Nebraska, to relative generalist, e.g., Rhabdias joaquinensis from Nebraska. Overall, host specificity for species of Rhabdias does not provide support for the evolution of progressive specialization over time. For most species of lungworms, host specificity in nature appears to be limited by both ecological and physiological factors, which vary between species and their hosts. Furthermore, some lungworms, e.g., Rhabdias bakeri from Missouri, appear to be tracking host resources instead of host phylogenies, an example of ecological fitting.

Spatial and temporal patterns of intraspecific morphological variation in Dactylogyrus simplexus from fathead minnows in Nebraska.

Dactylogyrus simplexus Monaco and Mizelle, 1955, occurs on the gills of fathead minnows (Pimephales promelas). Previous research on parasites of fathead minnows from 3 converging Nebraska streams, Elk Creek, Oak Creek, and West Oak Creek, shows that fish in each stream constitute distinct populations. To determine whether their parasites had diverged structurally in response to such isolation, or in response to seasonal change, we searched for patterns of intraspecific morphometric variation among D. simplexus. Over 3 collection dates in fall 2007, spring 2008, and fall 2009, 203 D. simplexus were collected from Elk and West Oak Creeks. We ran 1-way ANOVA to compare differences in 15 distinct point-to-point measurements of sclerotized parts across sites and collection dates. Significant differences were found in some D. simplexus measurements between Elk and West Oak Creeks for all 3 collection dates, but the characteristics that differed and the trend of variation between the creeks were not consistent over time. Dactylogyrus simplexus from both Elk and West Oak Creeks showed consistent patterns of variation over time for 5 measurements, including hamulus gap width, bar length, marginal hook length, sickle length, and sickle width. In conclusion, D. simplexus demonstrate consistent patterns of seasonal variation, but not spatial.

The role of damselflies (Odonata: Zygoptera) as paratenic hosts in the transmission of Halipegus eccentricus (Digenea: Hemiuridae) to anurans.

Halipegus eccentricus is a common hemiurid trematode in the eustachian tubes of North America frogs. However, the life cycle of this species has never been completely elucidated. Studies on H. eccentricus suggest that it has a 3-host life cycle. Here, we show through fieldwork and host specificity experimental infections that the life cycle of Halipegus eccentricus utilizes 4 hosts. Metamorphosed anurans become infected with H. eccentricus by feeding on infected damselflies; worms reside in the stomach of anurans, migrate to the eustachian tubes within 32-39 days post-exposure (DPE), and release eggs 50-60 DPE. Cystophorous cercariae develop in Physa gyrina snails within 32-35 DPE, infect ostracod ( Cypridopsis sp.) second intermediate hosts, and develop to metacercariae. Fifteen- to 19-day-old metacercariae from ostracods are infective to both damselfly larvae and metamorphosed anurans. Field surveys of damselflies and tadpoles, along with laboratory exposure of damselfly larvae, metamorphosed anurans, and tadpoles with infected ostracods, indicated that only metamorphosed anurans and damselflies become infected with H. eccentricus , whereas field-collected tadpoles and laboratory-exposed tadpoles were never infected with H. eccentricus . Because little morphological change occurred in the metacercaria stage of H. eccentricus between the ostracod second intermediate host and damselfly host, and metamorphosed anurans became infected with H. eccentricus metacercariae recovered from both host groups, we suggest that odonates serve as paratenic hosts in this life cycle. Additionally, our field work and experimental infections provide data on the use of odonates as the route of infection by another North American Halipegus sp. that matures in the stomach of frogs. Our data indicate that when the life cycles are known, the use of odonates as the route of infection to anurans is common in the life cycles of Halipegus spp., and all species exhibit remarkable infection site fidelity in their amphibian hosts.

Redescription of the frog bladder fluke Gorgoderina attenuata from the northern leopard frog, Rana pipiens.

Morphological characters used to differentiate North American bladder flukes, Gorgoderina spp., are problematic and different authors use different morphological characteristics for distinguishing species. More importantly, no type specimens exist for 4 of the 12 North American species infecting anuran and caudatan hosts. A redescription of Gorgoderina attenuata (Stafford, 1902) Stafford, 1905 is based on new collections from 6 species of anurans from Arkansas, Nebraska, New York, and Wisconsin. Morphological comparisons between gravid G. attenuata recovered from bullfrogs and northern leopard frogs indicated statistically significant differences in 11 of 28 morphological characters examined. However, there was overlap among all of these characters, and it is unclear whether these morphological types represent host-induced morphological changes in worm morphology or cryptic species. Based on our findings, we suggest that morphological and molecular data from G. attenuata-like worms recovered from the other 23 definitive hosts reported for G. attenuata need to be collected to resolve this issue.

Community structure and seasonal dynamics of Dactylogyrus Spp. (Monogenea) on the fathead minnow (Pimephales promelas ) from the Salt Valley Watershed, Lancaster County, Nebraska.

The gill monogene communities of Pimephales promelas (fathead minnow) in 3 distinct sites on converging streams were investigated from 2004 to 2006 in 3 different seasons. Thirty collections of P. promelas were made in southeastern Nebraska along 3 converging tributaries: Elk Creek (40.88534°N, 96.83366°W), West Oak Creek (40.9082°N, 96.81432°W), and Oak Creek (40.91402°N, 96.770583°W), Lancaster County, Nebraska. In all, 103 P. promelas were collected from Elk Creek, 115 from West Oak Creek, and 78 from Oak Creek and examined for gill monogenes. Among the P. promelas collected, 93.5% were infected with up to 3 species of Dactylogyrus, including Dactylogyrus simplex Mizelle, 1937, Dactylogyrus bychowskyi Mizelle, 1937, and Dactylogyrus pectenatus Mayes, 1977. Mean intensities at Elk Creek, West Oak Creek, and Oak Creek were 17.6, 22.8, and 25.1, and prevalences 88, 95, and 97%, respectively. At these 3 sites: (1) P. promelas does not share Dactylogyrus species with Semotilus atromaculatus (Creek chub) or Notropis stramineus (Sand shiner); (2) fish size and sex are not predictive of Dactylogyrus infection; (3) Dactylogyrus spp. vary (not always predictably) in their seasonal occurrence; (4) populations of Dactylogyrus spp. respond to environmental differences among sites; and (5) the community structure of Dactylogyrus spp. (order of abundance) is independent of environment.

Comparative life cycles and life histories of North American Rhabdias spp. (Nematoda: Rhabdiasidae): lungworms from snakes and anurans.

The present study used experimental infections to compare the life cycles and life histories of 6 Rhabdias spp. infecting snakes and anurans. Free-living development of anuran lungworms was primarily limited to heterogonic reproduction, and females utilized matricidal endotoky exclusively, whereas snake lungworms primarily reproduced homogonically and, when heterogonic reproduction occurred, females used a combination of releasing eggs and matricidal endotoky. Infective snake lungworms survived for longer periods in fresh water compared to anuran worms. Infective anuran lungworms penetrated into the skin of frogs and toads; few infections resulted from per os infections. In contrast, snake lungworms were unable to penetrate skin; instead, infective juveniles penetrated into snake esophageal tissue during per os infections. Despite separate points of entry, anuran and snake lungworms both migrated and developed in the fascia, eventually penetrating into the body cavity of the host. Worms molted to adulthood inside the body cavity and subsequently penetrated into the host's lungs, where they fed on blood while becoming gravid. Adult lungworm survival varied among lungworm species, but, in general, snake lungworms were longer lived than anuran worms. Anuran lungworms were poorly suited for transmission via transport hosts, whereas snake lungworms were consistently capable of establishing infections using transport hosts. Overall, these observations suggest that snake and anuran lungworms have discrepant life cycles and life history strategies.

Alternative life cycle strategies and colonization of young anurans by Gorgoderina attenuata in Nebraska.

Studies on life cycles and epizootiology of North American frog bladder flukes indicate that adult frogs become infected predominantly by ingesting tadpoles or other frogs that serve as second intermediate hosts for gorgoderid metacercariae. Other studies have indicated that newly metamorphosed frogs are rarely infected with these parasites because they are gape-limited predators that cannot feed on large intermediate hosts such as tadpoles and other frogs. We examined the role of potential intermediate hosts in the recruitment of the frog bladder fluke, Gorgoderina attenuata, to metamorphosed northern leopard frogs, Woodhouse's toads, and bullfrogs from western Nebraska. We completed the life cycle of G. attenuata in the laboratory in 3 anuran species by experimentally infecting a variety of hosts. In addition, we generated and compared DNA sequence data from life cycle stages collected from a variety of naturally infected hosts. Our field and laboratory data indicate that in Nebraska G. attenuata has a truncated, 2-host life cycle that includes fingernail clams and anurans. Cercariae are ingested directly by tadpoles; unencysted juvenile worms then develop in the kidneys of tadpoles before moving to, and maturing in, the urinary bladder when tadpoles metamorphose. Additionally, G. attenuata can infect metamorphosed leopard frogs, bullfrogs, and toads when metacercariae in damselfly second intermediate hosts are ingested. These worms can also infect adult bullfrogs when they feed on other infected anurans possessing worms in their kidneys. Comparison of our material to published accounts of G. attenuata morphology and life cycles in Massachusetts suggests that previous work may have inadvertently involved 2 different species of gorgoderids. Our comparative approach to life cycle studies in different anuran life stages and multiple species of hosts suggests that tadpoles and metamorphosed anurans have favored alternative life cycle strategies in this trematode.

Gregarines on a diet: the effects of host starvation on Gregarina confusa Janovy et al., 2007 (Apicomplexa: Eugregarinida) in Tribolium destructor Uyttenboogaart, 1933 (Coleoptera: Tenebrionidae) larvae.

This study was designed to explore the nutritional relationship between Gregarina confusa (Apicomplexa: Eugregarinida) parasites and its coleopteran host, Tribolium destructor, by measuring the cytoplasmic density of gregarines in continuously fed larvae, starved larvae, and larvae refed after starvation. Cultures were maintained in a standard media (whole wheat flour:commercial wheat germ:yeast [30:10:1]). Larvae from control and experimental groups were dissected daily for 3 days then allowed to feed or starve for an additional 3 days. On day 6, the remaining experimental larvae were divided and placed into 2 groups; 1 group remained starved while larvae from the second group were fed a Wheaties flake. Photographs were taken of the parasites daily and analyzed using ScionImage. Gregarines from starved larvae were significantly longer and skinnier than those from fed controls, and there was also a significant difference between gregarine deutomerite cytoplasmic densities. Parasites from refed larvae regained cytoplasmic density within 24 hr and showed morphological similarities to those from fed larvae. This study shows that the Tribolium destructor-Gregarina confusa relationship can be manipulated easily through alterations of host diet and thus is an excellent model for use in the study of chemical relationships between parasites and their hosts.

A tick from a prehistoric Arizona coprolite.

Ticks have never been reported in archaeological analyses. Here, we present the discovery of a tick from a coprolite excavated from Antelope Cave in extreme northwest Arizona. Dietary analysis indicates that the coprolite has a human origin. This archaeological occupation is associated with the Ancestral Pueblo culture (Anasazi). This discovery supports previous hypotheses that ticks were a potential source of disease and that ectoparasites were eaten by ancient people.

Evolutionary avenues for, and constraints on, the transmission of frog lung flukes (Haematoloechus spp.) in dragonfly second intermediate hosts.

Metacercariae survival patterns and their distribution in second intermediate odonate hosts were examined for 4 species of frog lung flukes. Surveys of aquatic larvae and recently emerged teneral dragonflies and damselflies indicated that prevalence and mean abundance of Haematoloechus spp. metacercariae were significantly lower in teneral dragonflies than larval dragonflies, while there was no significant difference in prevalence or mean abundance of Haematoloechus spp. metacercariae among larval and teneral damselflies. Experimental infections of dragonflies indicated that metacercariae of Haematoloechus coloradensis and Haematoloechus complexus were located in the head, thorax, and branchial basket of dragonflies, whereas metacercariae of Haematoloechus longiplexus and Haematoloechus parviplexus were restricted to the branchial basket of these hosts. Metacercariae of H. coloradensis, H. complexus, and H. longiplexus infected the head, thorax, and abdomen of damselflies, but these insects were resistant to infection with H. parviplexus. Subsequent metamorphosis experiments on experimentally infected dragonflies indicated that most metacercariae of H. longiplexus were lost from the branchial basket during metamorphosis, but most metacercariae of H. coloradensis, H. complexus, and H. parviplexus survived dragonfly metamorphosis. These observations suggest that the observed ecological host specificity of H. longiplexus in semiterrestrial leopard frogs may be due to few metacercariae of H. longiplexus reaching these frogs in a terrestrial environment. Because of the uncertain validity of Haematoloechus varioplexus as a distinct species from its synonym H. parviplexus, their morphological characters were reevaluated. The morphological data on H. varioplexus and H. parviplexus indicate that they differ in their acetabulum length and width, ovary shape, testes length, and egg length and width. Experimental infections of plains leopard frogs, northern leopard frogs, and bullfrogs with worms from bullfrogs indicate that the synonymy of H. parviplexus with H. varioplexus is not warranted, and that these flukes are distinct species, i.e., H. parviplexus in bullfrogs and H. varioplexus in plains leopard frogs and northern leopard frogs.

Small frogs get their worms first: the role of nonodonate arthropods in the recruitment of Haematoloechus coloradensis and Haematoloechus complexus in newly metamorphosed northern leopard frogs, Rana pipiens, and woodhouse's toads, Bufo woodhousii.

Studies on the life cycles and epizootiology of North American frog lung flukes indicate that most species utilize odonates as second intermediate hosts; adult frogs become infected by ingesting odonate intermediate hosts. Newly metamorphosed frogs are rarely infected with these parasites, predominantly because they are gape-limited predators that cannot feed on large intermediate hosts such as dragonflies. We examined the role of the frog diet and potential intermediate hosts in the recruitment of the frog lung fluke, Haematoloechus coloradensis, to metamorphosed northern leopard frogs (Rana pipiens), Woodhouse's toads (Bufo woodhousii), and bullfrogs (Rana catesbeiana) from western Nebraska. Because of the uncertain validity of H. coloradensis as a distinct species from Haematoloechus complexus, morphological characters of both species were reevaluated and the life cycles of both species were completed in the laboratory. The morphological data on H. coloradensis and H. coimplexus indicate that they differ in their oral sucker to pharynx ratio, uterine loop distribution, and placement of vitelline follicles. However, in terms of their life cycles, both species are quite similar in their use of physid snails as first intermediate hosts, a wide range of nonodonate and odonate arthropods as second intermediate hosts, and leopard frogs and toads as definitive hosts. These results indicate that H. coloradensis and H. complexus are generalists at the second intermediate host level and might be able to infect newly metamorphosed leopard frogs and toads by using small nonodonate arthropods more commonly than other frog lung fluke species. Comparisons of population structure of adult flukes in newly metamorphosed leopard frogs indicate that the generalist nature of H. coloradensis metacercariae enables it to colonize young of the year leopard frogs more commonly than other Haematoloechus spp. that only use odonates as second intermediate hosts. In this respect, the generalist nature of H. coloradensis and H. complexus at the second intermediate host level is an avenue for the colonization of young of year frogs.

A new species of Myxidium (Myxosporea: Myxidiidae), from the western chorus frog, Pseudacris triseriata triseriata, and Blanchard's cricket frog, Acris crepitans blanchardi (Hylidae), from eastern Nebraska: morphology, phylogeny, and critical comments on amphibian Myxidium taxonomy.

During March 2001-April 2004, 164 adult anurans of 6 species (47 Rana blairi, 35 Rana catesbeiana, 31 Hyla chrysoscelis, 31 Pseudacris triseriata triseriata, 11 Bufo woodhousii, and 9 Acris crepitans blanchardi) from Pawnee Lake, Lancaster County, Nebraska, were surveyed for myxozoan parasites. Of these, 20 of 31 (65%) P. triseriata triseriata and 1 of 9 (11%) A. crepitans blanchardi were infected with a new species of Myxidium. Myxidium melleni n. sp. (Myxosporea) is described from the gallbladder of the western chorus frog, P. triseriata triseriata (Hylidae). This is the second species of Myxidium described from North American amphibians. Mature plasmodia are disc-shaped or elliptical 691 (400-1,375) x 499 (230-1,200) x 23 (16-35) microm, polysporic, producing many disporic pansporoblasts. The mature spores, 12.3 (12.0-13.5) x 7.6 (7.0-9.0) x 6.6 (6.0-8.0) microm, containing a single binucleated sporoplasm, are broadly elliptical, with 2-5 transverse grooves on each valve, and contain 2 equal polar capsules 5.2 (4.8-5.5) x 4.2 (3.8-4.5) microm positioned at opposite ends of the spore. Myxidium melleni n. sp. is morphologically consistent with other members of Myxidium. However, M. melleni n. sp. was phylogenetically distinct from other Myxidium species for which DNA sequences are available. Only with improved morphological analyses, accompanied by molecular data, and the deposit of type specimens, can the ambiguous nature of Myxidium be resolved. Guidelines for descriptions of new species of Myxidium are provided.

Observations on myiasis by the calliphorids, Bufolucilia silvarum and Bufolucilia elongata, in wood frogs, Rana sylvatica, from southeastern Wisconsin.

Larvae of certain species of blowflies (Calliphoridae) can cause myiasis in frogs and toads, but there are few reports from North American amphibians. Of these, most are from toads (bufonids). In this study, we observe primary myiasis in a population of juvenile wood frogs, Rana sylvatica, collected on 22-23 August 2003, from southeastern Wisconsin and compare our observations with previous studies on myiasis from toads. Two (5%) of 39 frogs were infected by the blow fly Bufolucilia silvarum, with an intensity of 28 and 31, whereas 1 (2.5%) of 39 frogs was infected by the blow fly Bufolucilia elongata with an intensity of 14. We found that (1) B. silvarum lay eggs on healthy wood frogs, (2) eggs hatch, with first-instar maggots penetrating under the skin, (3) maggots develop to mature third instars within 13-16 hr of egg hatching, (4) maggots kill the host within 7-47 hr of egg hatching, and (5) maggots consume the entire frog carcass reducing it to bones within 42-59 hr of egg hatching. Our observations on the time of death and how quickly carcasses of wood frogs were consumed by these maggots compared with previous studies on toads suggest that finding infected juvenile wood frogs may be uncommon. Therefore, myiasis by these flies on wood frogs and other small terrestrial anurans may be a phenomenon that is much more common than is currently observed. This is the first report of B. silvarum and B. elongata causing myiasis in wood frogs.

Observations on the life history and descriptions of coccidia (Apicomplexa) from the western chorus frog, Pseudacris triseriata triseriata, from eastern Nebraska.

Two hundred and twenty-four anurans of 6 species (47 adults and 16 tadpoles of Rana blairi, 35 R. catesbeiana, 31 Hyla chrysoscelis, 30 adults and 46 tadpoles of Pseudacris triseriata triseriata, 11 Bufo woodhousii, and 8 Acris crepitans) from Pawnee Lake, Lancaster County, Nebraska, were surveyed for coccidian parasites during March 2001 to May 2002. Of these, 23 of 30 (77%) adults and 4 of 46 (9%) tadpoles of P. t. triseriata shed oocysts of Isospora cogginsi n. sp. Oocysts of I. cogginsi were ovoid, 19.3 x 15.1 (18-23 x 11-20) microm, with a thin, smooth, colorless, single-layered wall, with no micropyle or oocyst residuum. Sporocysts were ovoid, 13.3 x 9.9 (11-15 x 9-13) microm, with a thin, colorless, smooth wall, and Stieda body absent. Sporocyst residuum was present, 5.5 x 5.3 (4-7 x 4-7) microm, consisting of numerous granules. Histological examination of frogs and tadpoles infected with the new species revealed endogenous stages including mature meronts, developing microgamonts, mature microgametes, mature macrogamonts, and young unsporulated oocysts located in the cytoplasm of the epithelial cells of the small intestine. Concurrently, 2 adult P. t. triseriata shed oocysts of Eimeria streckeri. Oocysts of E. streckeri were spherical, 15.7 x 15.4 (14-17 x 14-19) microm, with a thin, smooth, single-layered, colorless wall with an oocyst residuum composed of numerous granules surrounding a large vacuolated area, with a previously undescribed globularlike body present within the vacuole, and no micropyle. Sporocysts were ovoid, 9.1 x 6.1 (7-10 x 5-7) microm, with a thin, colorless, smooth wall with a Stieda body and sporocyst residuum. Our results are the first to document infection of adult and tadpole stages of frogs of the same species with the same species of coccidian, indicating that adult frogs may contaminate breeding ponds with oocysts during their breeding season and infect tadpoles directly by the ingestion of sporulated oocysts.

Phyllodistomum funduli n. sp. (Trematoda: Gorgoderidae) from Fundulus sciadicus Cope from Cedar Creek in western Nebraska.

Phyllodistomum funduli n. sp. is described from the urinary bladder and ureters of the plains topminnow, Fundulus sciadicus, from Cedar Creek in Keith County, Nebraska (41 degrees 11.18'N, 101 degrees 21.77'W). Phyllodistomum funduli differs from most other Phyllodistomum species reported in possessing an oral sucker that is larger than the acetabulum. Currently, 7 other species of Phyllodistomum have been reported to possess this trait. A comparison of P. funduli with these 7 other species indicates that P. funduli is a distinct species, lacking notches, posterior body folds, caudal projections, or cephalic glands found in some congeners. There is no published report on parasites other than monogeneans inhabiting the plains topminnow, and there is no other report of members from the Gorgoderidae inhabiting any members of the Fundulidae.