A site-specific glycosylation of salivary glands (SGs) isolated from unfed and partially fed Ixodes ricinus females was identified with the use of lectin affinity labeling on sections and western blots of SDS-PAGE gels. The results revealed that secretory granules of a, b, and c cells of the type II acinus and e and f cells of the type III acinus are glycosylated. In partially engorged tick SGs, 2 subtypes of c cells were distinguished. The granules of c1 cells contained mannose, N-acetyl-d-glucosamine, and sialic acid residues. The granules of b, c2, and e cells exhibited complex glycoconjugates rich in mannose, N-acetyl-D-glucosamine, galactose, N-acetyl-d-galactosamine, and a moderate amount of sialic acid. The granules of f cells contained N-acetyl-d-glucosamine and mannose moieties. Type III acini surfaces were covered with mannose-specific ConA binding sites. Except the granules of salivary cells, sialic acid–specific lectins MAA II and SNA strongly bound cuticular structures of alveolar ducts, and weakly with the cuticular spiral thread of excretory salivary ducts. The total sialic acid level in SG homogenates isolated from partially fed females was determined by the thiobarbituric acid method. Sialic acid, which has been found during the development of a few insect species, has not been reported in ticks as yet.

Ultrastructure of spermiogenesis and the main characters of the mature spermatozoon of Troglotrema acutum are described by means of transmission electron microscopy. Specimens were obtained from the nasolacrimal sinuses of an American mink (Mustela vison). Spermiogenesis in T. acutum follows the general pattern of digeneans. The zone of differentiation is a conical-shaped area bordered by cortical microtubules and delimited at its base by a ring of arched membranes. This area contains 2 centrioles associated with striated rootlets and an intercentriolar body between them. The centrioles develop 2 free flagella that grow ortogonally to the median cytoplasmic process. The posterior flagellar rotation and proximodistal fusion of the free flagella with the median cytoplasmic process originate the spermatozoon. The mature spermatozoon of T. acutum is characterized by the presence of 2 axonemes of different lengths presenting the 9 ‘1’ trepaxonematan pattern, 2 bundles of parallel cortical microtubules, 2 mitochondria, a nucleus, and granules of glycogen. These ultrastructural characters are compared with other digenean species previously studied and the importance of different spermatological features is discussed.

We studied the influence of host age and sex on the helminth fauna of 324 Larus michahellis captured in different locations in the region of Galicia (northwestern Spain). Gulls were grouped into prefledglings, first-year immature birds, second- and third-year immature birds, and adults. Second-year, third-year, and adult birds were grouped by sex. Thirty-six helminth species were recorded. Total species richness and mean infracommunity species richness were both significantly lower for prefledglings than for the other age groups. Prevalence increased significantly with age for Brachylecithum microtesticulatum, probably reflecting changing feeding habits. Likewise, 8 species (Cardiocephaloides longicollis, Microphallus similis, Maritrema gratiosum, Gynaecotyla longiintestinata, Brachylecithum microtesticulatum, Himasthla elongata, Parorchis acanthus, and Renicola sp.) were absent or had very low prevalence in prefledglings. At least 5 of these 8 species are transmitted to gulls through ingestion of molluscs or crustaceans, which suggests that these types of prey are seldom fed to prefledglings. In Gymnophallus deliciosus, G. longiintestinata, and Cosmocephalus obvelatus, mean intensity, and in the latter case prevalence, declined with age, suggesting that protective immunity against these species increase with age. Only G. deliciosus, Microphallus similis, and G. longiintestinata presented significant differences between the sexes.

Viviparous gyrodactylids are remarkable monogenoid ectoparasites, not only because of their speciousness, but also due to their unusually wide range of hosts. Although many factors have been proposed to determine the location where gyrodactylids attach to their hosts, little is known about how their preference for specific host body regions changes over the course of infection. In this study, we investigate the dynamics of topographical specialization of the parasite Gyrodactylus anisopharynx on 2 of its natural freshwater fish hosts (Corydoras paleatus and C. ehrhardti), as well as a naïve host (C. schwartzi). We recorded the spatial location of this parasite from the foundation of the infrapopulation to its extinction to assess how topographical specialization is affected by host species, the size and the age of the infrapopulation, and the possibility of transmission among hosts. Our results indicate that topographical specialization is negatively correlated with infrapopulation size and only marginally affected by infrapopulation age. Also, the degree of specialization was different among host species, but seemed unaffected by the possibility of transmission among hosts. Therefore, observed changes in spatial specialization of G. anisopharynx do not appear to represent adaptive responses to maximize their transmission. Rather, mechanisms such as increased competition and/ or local immune responses might cause parasites to occupy less favorable regions of the body of their hosts with increasing density.

Leptorhynchoides thecatus (Linton, 1891), an acanthocephalan parasite of freshwater fishes, varies in host use, development, and habitat use throughout North America. Spatial structure of these characteristics was examined from data extracted from the literature. Geographic patterns were inferred from point comparisons using correllograms and then tested with Moran's I statistic for global and local significance, and visually from regional means within major river drainages. Species of Micropterus Lacepède, 1802 (black basses) were common hosts in most regions, except the Lower Mississippi and South Atlantic regions where species of Lepomis Rafinesque, 1819 (sunfishes) were common hosts. Development, described as the proportions of adults relative to cystacanths (extraintestinal juveniles), decreased with latitude. Habitat use of L. thecatus showed marked geographic patterns. Leptorhynchoides thecatus occurred in the intestine of sunfishes in the South Atlantic and Lower Mississippi regions, in the ceca in fish of all species included in the study in the Missouri and Texas-Gulf regions, and both in ceca and intestines in fish of all species in northern regions. Leptorhynchoides thecatus showed geographic patterning within the variable traits across the range of the species. These patterns may be the result of ecological factors or of genetic differences that might indicate L. thecatus comprises multiple cryptic species.

Infections with sea lice species belonging to Lepeophtheirus and Caligus are reported from examinations of 1,309 three-spine sticklebacks collected in coastal British Columbia. Over 97% of the 19,960 Lepeophtheirus specimens and nearly 96% of the 2,340 Caligus specimens were in the copepodid and chalimus developmental stages. The parasites were identified as Lepeophtheirus salmonis and Caligus clemensi based on morphology of adult stages. Between 1,763 and 1,766 base pairs (bp) of 18S rDNA from adult specimens collected from sticklebacks and salmon differed from the GenBank L. salmonis reference sequence by a single bp and were distinct from those of 2 other Lepeophtheirus species. A 530-bp region of 18S rDNA from chalimus stages of Lepeophtheirus obtained from sticklebacks and salmon was identical to that of the L. salmonis reference sequence. The three-spine stickleback is a new host record for L. salmonis. The prevalence of L. salmonis was 83.6% and that of C. clemensi was 42.8%. The intensities of these infections were 18.3 and 4.2, respectively. There was no significant relationship between sea lice abundance and stickleback condition factor. Significant spatial and temporal variations both in abundance of sea lice and surface seawater salinities were measured. The abundance of both sea lice species was lowest in zones in which surface seawater salinity was also lowest. Sticklebacks appear to serve as temporary hosts, suggesting a role of this host in the epizootiology of L. salmonis. The stickleback may be a useful sentinel species with which to monitor spatial and temporal changes in the abundance of L. salmonis and C. clemensi.

Cornutohaptor nigrescensi n. sp. (Diplectanidae) is described from the gills of the black snook, Centropomus nigrescens (Perciformes: Centropomidae) from the Pacific coast of Mexico. Cornutohaptor n. gen. is proposed for this new species and is characterized by possessing 2 intestinal ceca terminating blindly; a germarium looping right intestinal cecum; bilobed testis; 2 seminal vesicles; 7 pairs of hooks, each with protruding thumb; a grooved ventral bar and coiled male copulatory organ (MCO); an accessory piece comprising a “baglike structure” with an appendage; dorsal bars associated parallelly to body midline; and no adhesive accessory organs on the haptor. Cornutohaptor differs from all confamilial genera by including species with anchors with straight and deep root longest, hook pair 1 reduced in size, MCO with counterclockwise rings, and by the morphology of the accessory piece. Cornutohaptor nigrescensi most closely resembles species of Murraytrema Price, 1937, Lobotrema Tripathi, 1937, and Murraytrematoides Yamaguti, 1958, because of the absence of squamodiscs or lamellodiscs on the haptor and tegumental scales on the posterior portion of the body. Cornutohaptor differs from these genera in the position and number of haptoral bars (2 bars in Lobotrema spp., dorsal bars transversally associated in Murraytrema and Murraytrematoides spp.) and in having a coiled MCO (copulatory organ is a comparatively straight, poorly sclerotized tube in Murraytrematoides spp.). This is the first diplectanid described from a centropomid along the Pacific coast of Mexico.

A comparative morphological study of specimens of Ligophorus spp. from Mugil cephalus in western Mediterranean and the Black Sea localities has been carried out, indicating the presence of 2 distinct forms, i.e., Ligophorus chabaudi and Ligophorus cephali n. sp. A detailed description of the latter and an up-to-date redescription of L. chabaudi are provided. The existence of these 2 morphological species was additionally supported by principal component analysis based on 19 metric characters of 87 specimens arranged in samples defined by parasite species and geographical locality. The analysis indicated consistent differences between species but not between localities; this pattern was well supported by jackknife procedures. Linear discriminant analyses showed that the main metric differences between the 2 species were attributable to the lengths of the dorsal and ventral bars and to the uncinulus length; use of the first 2 variables was enough to allocate all specimens studied to either form. The erection of L. cephali n. sp. raises the number of sympatric Mediterranean species of Ligophorus on M. cephalus to 3. The known geographical ranges of these species are compared.

Schistosoma mansoni belongs to the dioecious Schistosomatidae. The occasional observation of males with rudimentary female characteristics is thought to attest the hermaphroditic roots of this parasite. Supernumerary testicular lobes also are recurrently seen in this helminth, but their morphology and origin are elusive. Here, we investigated the morphology of the supernumerary lobes from 15 S. mansoni males and similar structures of 2 females in whole mounts by brightfield and confocal laser scanning microscopy. The follicles in the females were not regarded as testicular lobes, but 1 male had a rudimentary ovary with a residual oviduct posterior to the normal set of testicular lobes and was considered hermaphroditic. In addition, 6 other males contained supernumerary lobes that enclosed fully matured oocytes in variable amounts. In the remaining 8 males, no female characteristics could be seen, and the lobes showed morphologically a more testicular aspect, although this morphology was only confirmed in 1 male where spermatozoids were detected. The process underlying the expression of supernumerary testes or lobes with oocytes is similar and is, at least in part, induced by the host.

Pygidiopsis pindoramensis Travassos, 1928, is redescribed from the holotype and specimens obtained from experimental infections of hamsters with metacercariae of the naturally infected poeciliids Poecilia vivipara and Phalloptychus januarius from Rio de Janeiro, Brazil. Ascocotyle (Phagicola) pindoramensis (Travassos, 1928) n. comb. is proposed, based on the presence of a solid, tapering muscular posterior prolongation of the oral sucker and Ascocotyle (Phagicola) mollienisicola (Sogandares-Bernal and Bridgman, 1960) is synonymized with A. (P.) pindoramensis because of the identical morphology and measurements of adults and metacercariae, similar spectrum of fish intermediate hosts (poeciliids), and the same site of infection of the metacercariae. Trematodes designated as Pygidiopsis pindoramensis, previously reported from Argentina and Mexico, represent another species of Pygidiopsis Looss, 1907, because they possess circumoral spines and their oral sucker is devoid of the posterior muscular prolongation. Data on the surface morphology of metacercariae and adults of A. (P.) pindoramensis are inferred from scanning electron microscopy observations.

We studied variation in isozyme patterns of 8 metabolic enzymes in 5 species of Heterorhabditis (H. bacteriophora, H. indica, H. marelata, H. megidis, and H. zealandica) comprising 18 isolates. Isozyme banding patterns of all the 8 enzymes were species specific; however, 3 enzymes, i.e., arginine kinase, fumarate hydratase, and malate dehydrogenase, displayed distinct patterns among all the 18 isolates. Phylogenetic analysis of the isozyme patterns produced dendrograms depicting a high degree of genetic variation among Heterorhabditis species, with the average pairwise distance of 0.2000. Trees constructed using different phylogenetic methods showed a relatively close genetic relationship between H. megidis and H. zealandica and between H. bacteriophora and H. indica. Also, H. bacteriophora HP88 was the most distant species from H. megidis (UK isolate), H. marelatus (Oregon isolate), and H. zealandica (X1 isolate) with pairwise distance of 0.1957, 0.2228, and 0.2120, respectively. Phylogenetic analysis also revealed genetic variation among H. bacteriophora isolates with the average pairwise distance of 0.1507. GPS2 and GPS3 were the most closely related isolates with the average distance of only 0.0870, followed by GPS1 and GPS2 with average distance of 0.1087. In contrast, KMD19 and HP88, OH25, and HP88, and OH25 and Acows isolates were the most divergent populations with a pairwise distance of 0.2011 and 37 character differences. Pairwise distance analysis also revealed that genetic divergence among populations of H. bacteriophora is relatively independent of geographic distance. Overall, these results demonstrate strong subspecies structuring in H. bacteriophora.

Neospora caninum is a world-wide parasite that causes neuromuscular disorders in dogs and bovine abortion. Biological diversity among isolates has been proved in both in vivo and in vitro studies. In contrast, little is known about the genetic diversity of this parasite. Microsatellite sequence analysis constitutes a suitable tool that has been used for the genetic analysis of other apicomplexan parasites. In this report, we describe the identification and analysis of 13 microsatellite loci from N. caninum DNA sequences deposited in public databases, which were evaluated with the use of 9 isolates grown in vitro. One microsatellite was monomorphic, and the remaining 12 loci exhibited 3 to 9 separate alleles. Multilocus analysis showed that each of the 9 isolates investigated here displayed a unique profile and revealed no association between the genetic similarity and host or geographic origin. The multilocus analysis approach described here might nevertheless provide the powerful tool needed to study the genetic complexity of N. caninum and the molecular epidemiology of neosporosis.

Partial sequences of mitochondrial genes nad1 (316 bp) and cox1 (429 bp) were analyzed to estimate the variability of the liver fluke samples collected in 20 localities in Russia, Belarus, Ukraine, Bulgaria, Armenia, Azerbaijan, Georgia, Turkey, Turkmenistan, and China. The sequences had 4.1% (nad1) and 2.3% (cox1) of variable sites, and 13 and 10 haplotypes were identified among nad1 and cox1 genes, respectively. Spatial analysis of genetic and nucleotide diversity indicated little or no structuring of genetic variation between hosts or regions. The analysis of distribution of both separate and combined (nad1 cox1) haplotypes revealed the existence of 2 well-defined lineages with 2 main haplotypes and a number of shared divergent haplotypes. Our study showed that the first lineage included the main N1–C1 haplotype, which was found in Australia, China, Georgia, Turkey, Armenia, Azerbaijan, and in all European populations (from Russia, Belarus, Ukraine, Bulgaria). The second lineage was found in all European populations and in populations from Armenia and Azerbaijan. It was suggested that one of the lineages (I) has an Asian origin. The possible source of mtDNA variability and associations between lineage divergence of parasite and its definitive hosts (cattle and sheep) are discussed.

Mice were fed ad libitum with a normal diet (25% protein) or low-protein diets (0–12.5% protein) for a wk and then infected with a nonlethal or lethal strain of Plasmodium yoelii, that is, blood stage infection. The same diet was continued until recovery. Mice fed with a normal diet showed severe parasitemia during nonlethal infection, but survived the infection. They died within 2 wk in the case of lethal infection. However, all mice fed with low-protein diets survived without apparent parasitemia (there were small peaks of parasitemia) in cases of both nonlethal and lethal strains. These surviving mice were found to have acquired potent innate immunity, showing the expansion of NK1.1⁻TCR^int cells and the production of autoantibodies during malarial infection. Severe combined immunodeficiency (scid) mice, which lack TCR^int cells as well as TCR^high cells, did not survive after malarial infection of lethal strain of P. yoelii, even when low-protein diets were given. These results suggest that low-protein diets enhanced innate immunity and inversely decreased conventional immunity, and that these immunological deviations rendered mice resistant against malaria. The present outcome also reminds us of our experience in the field study of malaria, in which some inhabitants eventually avoided contracting malaria even after apparent malarial infection.

Cystacanths of Oncicola venezuelensis (Acanthocephala: Oligacanthorhynchidae) were discovered in the hemocoel of Caribbean termites (Nasutitermes acajutlae) on St. Thomas and St. John islands in the U.S. Virgin Islands. In addition to occurring in the insect intermediate host, cystacanths were present in subcutaneous nodules of lizards (Anolis cristatellus and Anolis stratulus), in the greater omentum of small Indian mongooses (Herpestes auropunctatus), and embedded in mesenteries of pearly-eyed thrashers (Margarops fuscatus). These vertebrates likely are paratenic hosts, although a definitive host in the Virgin Islands is yet to be discovered. Cystacanths from intermediate and paratenic hosts agree fully with the original description of proboscis armature, including size and shape of hooks and their roots, of the species. Qualitative features of developing and growing structures agree with the original description of the species, but the sizes are smaller.

Cercariae of many digenean trematodes target particular developmental stages of their hosts. For some digeneans that are parasites of amphibians, infection appears timed to host metamorphosis. The success and timing of metamorphosis is itself affected by a number of factors, including host density. We investigated the degree to which rearing density of Rana pipiens larvae influenced time to metamorphosis and snout-to-vent length and mass at metamorphosis, as well as establishment of cercariae of the trematode Manodistomum syntomentera Stafford, 1905. As expected, individuals metamorphosed later, were smaller, and weighed less at metamorphosis if they were reared under intermediate to high densities compared with low densities, in experimental outdoor mesocosms. Cercariae establishment was higher in smaller metamorphs that took longer to metamorphose within the low-density treatment. Additionally, cercariae establishment was lower in larvae from the low-density tanks compared with larvae from the intermediate- to high-density tanks. However, more tadpoles had failed to metamorphose in the intermediate to high rearing densities by the time cercariae were no longer available from natural collections of first intermediate hosts, Physa spp. Larval amphibians under crowded conditions should experience increased susceptibility to trematode establishment in nature, but only if they metamorphose within the time period when cercariae are still available.

The possession of genital spines has been considered as a key taxonomic trait to differentiate Corynosoma from other genera of the Polymorphidae. However, Corynosoma currently consists of 2 groups of species with clear ecological and morphological divergences: the “marine” group (with ca. 30 species) infects mammals and piscivorous birds in the marine realm, whereas the “freshwater” group (with ca. 7 species) infects waterfowl in continental waters. Species from these groups differ in shape of body and neck, trunk spination, lemnisci length and shape, testes arrangement, and number and shape of cement glands. We tested whether species from these 2 groups formed a monophyletic assemblage based on a phylogenetic analysis by using 15 morphological characters. We also included species of Andracantha, Polymorphus, and Hexaglandula with which potential taxonomic conflicts could most likely arise. We obtained 108 equally most parsimonious trees of 32 steps, with a consistency index (CI) = 0.59, and a retention index (RI) = 0.82. The strict consensus tree indicated that the “freshwater” species of Corynosoma form a monophyletic assemblage closely related to some species of Polymorphus, whereas the “marine” species of Corynosoma are grouped together with Andracantha. Accordingly, Corynosoma is not a monophyletic assemblage, and Pseudocorynosoma n. gen. is proposed for the “freshwater” species of Corynosoma. This decision was strongly supported by (1) a functional comparison of foretrunk muscles between species of Polymophus, Andracantha, and Corynosoma; (2) a multivariate morphometric study of proboscis characters and egg size; and (3) an analysis of ecological patterns of host–parasite relationships.

A new species of allocreadiid digenean is described from Priapichthys annectens (Regan) (Osteichthyes: Poeciliidae) in the Área de Conservación Guanacaste, Costa Rica. A new genus, Paracreptotrema, is proposed to accommodate this species as well as Fellodistomum mendezi Songandares-Bernal, 1955, which was previously described from another poeciliid, Brachyrhaphis episcopi, in Panama. Paracreptotrema differs from all other nominal genera of Allocreadiidae by a combination of its symmetrical testes, restricted vitellaria, and the lack of oral lappets (muscular ‘papillae’) or other such appendages. Paracreptotrema blancoi n. sp. resembles Creptotrema creptotrema Travassos, Artigas and Pereira, 1928, but differs in lacking ventral oral lappets and in having vitellaria extending posteriorly only to the level of the testes. It can be distinguished from P. mendezi n. comb. in having a relatively larger and more posteriorly placed acetabulum, vitellaria that are more restricted anteriorly, smaller testes, and a uterus that does not extend to the posterior end of the body. The genus seems to be restricted to poeciliids in Central America.

Xiphocephalus triplogemmatus n. sp. and Xiphocephalus quadratogemmatus n. sp. (Apicomplexa: Eugregarinida) are described from Eleodes tricostata and Eleodes fusiformis (Coleoptera: Tenebrionidae), respectively, collected from Keith County in the sandhills of western Nebraska. Gamonts can be diagnosed with some confidence based on relative size and shape of the deutomerite, but these taxa are distinguished by differences in oocyst size, shape, and residuua number. Together with Xiphocephalus ellisi from Eleodes opacus in the same region, X. triplogemmatus and X. quadratogemmatus form a distinct Nearctic xiphocephalid group that is morphologically distinct from groups that occur in the Palearctic, Ethiopian, and Oriental regions.

A new Himasthlinae species, Curtuteria arguinae, is described as metacercariae from the cockle Cerastoderma edule (L.), collected at Banc d'Arguin (southwestern France). These metacercariae encysted preferentially in the mantle and also in the foot of cockles. Encysted and chemically excysted metacercariae were studied by light microscopy and scanning electron microscopy, respectively. Excysted metacercariae were elongated and curved ventrally. They bore a 33-spine circumoral collar. Sensory papillae were arranged around the oral sucker and also symmetrically along the ventral surface body, from the collar to the acetabulum. The dorsal and ventral tegument surfaces were densely packed with similar pointed spines. The posterior end of the body was without any spines. Among the Curtuteria species described previously, only Curtuteria haematopodis Smogorjewskaja and Iskova, 1966 had the same number of circumoral collar spines. A 6-yr field survey showed that the cockle population at Banc d'Arguin was subjected to a summer infection of C. arguinae. Curtuteria arguinae phenology of infection is characterized by interannual variability and seasonality (beginning in July–August and maximum in autumn). The first intermediate and final hosts remain unknown.

The lungs of a northern elephant seal, Mirounga angustirostris (Gill, 1866), and Pacific harbor seal, Phoca vitulina richardii (Gray, 1864), from the eastern central Pacific, central California coast, were examined for parasites. This represents the first thorough description and identification of Parafilaroides species from northern elephant seals and Pacific harbor seals on the west coast of North America. From this study, 2 new species of Filaroides (Parafilaroides) are described, illustrated, and differentiated by shape of vaginal sphincter, body morphometrics, and spicule shape from the 4 existing, recognized species in the subgenus. In addition, the subgenus Parafilaroides is proposed to be reestablished to full generic status in the Filaroididae based on the presence of caudal papillae and DNA.

Vexillata geomyos n. sp. is described as a parasite of Attwater's pocket gopher, Geomys attwateri (Geomyidae), from Welder Wildlife Refuge, Texas. The new species possesses 12 cuticular ridges; based on this character, it can be distinguished from 7 of the 13 species in the genus. From the 6 remaining species, V. geomyos can be differentiated because the dorsal ray in 3 of them (V. armandae, V. dessetae, and V. vexillata) has 2 short branches arising from main trunk, whereas dorsal ray of the new species lacks these branches. In addition, V. geomyos can be distinguished from V. brooksi because its spicules are divided at the tip (whereas in the new species, spicules are whole). The new species differs from V. legallae and V. convoluta in the number and arrangement of cuticular ridges at the posterior region of the body.

Sciurodendrium bravohollisae n. sp. (Heligmonellidae) is described as an intestinal parasite of 2 squirrels, Sciurus aureogaster Cuvier, 1829, collected from Los Robles, Municipio de Tlalnepantla, Morelos State, Mexico. The new species differs from all other congeners in possessing a very well-developed and sacciform genital cone. This is the seventh species described for the genus and the first one recorded in Mexico.

A bulk analysis of inter-simple sequence repeat–polymerase chain reaction (ISSR-PCR) provides a quick, reliable, and highly informative system for DNA banding patterns that permit species identification. The present study evaluates the applicability of this system to Trichinella species identification. After a single amplification carried out on a single larva with the primer 816([CA]_nRY) under high stringency conditions, which provide high reproducibility, we were able to identify by consistent banding patterns 5 sibling species: Trichinella spiralis (ISS48), 2 Trichinella britovi isolates (ISS11 and ISS86), Trichinella murrelli (ISS35), Trichinella nativa (ISS71), Trichinella nelsoni (ISS29); 3 additional Trichinella genotypes: T8 (ISS149), T9 (ISS408 and ISS409), and T6 (ISS34); and the nonencapsulated species Trichinella pseudospiralis (ISS13). Moreover, 33 new Trichinella isolates from 2 zoogeographical regions were unequivocally identified. All Trichinella isolates have shown an identical pattern with those produced by the reference strain. According to these data, we have demonstrated that ISSR-PCR is a robust technique that emerges as a useful new application for the molecular identification of Trichinella isolates in epidemiological studies.

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) × 499 (230–1,200) × 23 (16–35) μm, polysporic, producing many disporic pansporoblasts. The mature spores, 12.3 (12.0–13.5) × 7.6 (7.0–9.0) × 6.6 (6.0–8.0) μm, 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) × 4.2 (3.8–4.5) μm 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.

Megasolena dongzhaiensis n. sp. was collected from the intestine of Scatophagus argus (Linnaeus) (Perciformes: Scatophagidae) from the Dongzhai Bay (110°32′–37′E, 19°51′–20°1′N), Hainan Province, China. It resembles Megasolena acanthuri Machida and Uchida, 1991 in having larger body size, but it differs from the latter species in having an oral sucker that is larger, instead of smaller, than the acetabulum (sucker length ratio 1:0.456–0.494, and width ratio 1:0.61–0.65 in M. dongzhaiensis n. sp., as opposed to 1:1.3–1.7 in M. acanthuri). Moreover, the cuticle is spinose rather than aspinose; there is a band of circular muscle in the pharynx, and its eggs are smaller instead of larger (0.062–0.068 × 0.036–0.039 in M. dongzhaiensis n. sp., compared with 0.087–0.103 × 0.058–0.072 in M. acanthuri). Finally, the intestinal bifurcation is anterior, instead of dorsal, to the acetabulum. It resembles other species of Megasolena Linton, 1910 in having oral sucker larger than the acetabulum, and in having a circular muscle band in the oral sucker and pharynx, but it differs in having a larger body and smaller eggs. This is the first record of a Megasolena species from ray-finned fishes as well as in China.

There are many reports of cryptosporidial infection in ostriches, but none with molecular characterization of the isolates. A study was undertaken for the characterization of a Brazilian Cryptosporidium sp. ostrich isolate by using molecular phylogenetic analysis of fragments of the 18S ribosomal DNA, heat-shock protein (hsp) 70 coding gene, and actin coding gene. Biological studies were accomplished by the experimental inoculation of chickens via oral or intratracheal routes with fresh ostrich Cryptosporidium sp. oocysts. Molecular analysis of nucleotide sequences of the 3 genes by using neighbor-joining and parsimony methods grouped the ostrich isolate as a sister taxon of Cryptosporidium baileyi and showed that the ostrich isolate is genetically distinct from all other known Cryptosporidium species or genotypes. None of the inoculated chickens developed infection as determined by mucosal smears, histology, and fecal screening for oocysts. Although biological and molecular studies indicate that the ostrich Cryptosporidium is a new species, further studies regarding morphological, biological, and molecular characteristics of other ostrich isolates are required to confirm the species status of the ostrich Cryptosporidium.

An overlooked reference led to the production of an homonym and the correct type host for Leucocytozoon bennetti is the European Roller, Coracias garrulas. The emended name proposed from the hook-billed vanga, Vanga curvirostris, is renamed L. vangis n. nov.

Capillaria (Procapillaria) navoneae n. sp. is described from the congrid fish Conger orbignianus from waters off Mar del Plata, Argentina (38°08′S, 57°32′W) (prevalence 29.17%, mean intensity ± SD 6.07 ± 6.59). The new species is assigned to the subgenus Procapillaria Moravec, 1987, despite the absence of vulvar appendages in gravid females. Of the 3 species so far known in the subgenus, the new one most closely resembles C. (P.) margolisi, from which it is distinguished by lacking a medial expansion in the spicule and a slightly elevated anterior lip of vulva, and by having a shorter esophagus, a larger number of stichocytes, and a shorter tail.

Rhabdias bakeri n. sp. is described from specimens found in lungs of the wood frog, Rana sylvatica, from North Dakota. The new species has previously been mistakenly identified as Rhabdias ranae Walton, 1929, a common parasite of the leopard frog, Rana pipiens. The new species differs from R. ranae and Rhabdias joaquinensis Ingles, 1935 by the shape and size of pseudolabia, shape and size of buccal capsule, and wider esophageal bulb. Molecular analysis based on the partial sequences of nuclear 18S rDNA gene, complete sequences of internal transcribed spacer region, and partial sequences of 28S gene demonstrates clear differences between Rhabdias from Ra. sylvatica and Ra. pipiens, and supports the status of R. bakeri as a new species.

The ability of ponazuril to prevent or limit clinical signs of equine protozoal myeloencephalitis (EPM) after infection with Sarcocystis neurona was evaluated. Eighteen horses were assigned to 1 of 3 groups: no treatment, 2.5 mg/kg ponazuril, or 5.0 mg/kg ponazuril. Horses were administered ponazuril, once per day, beginning 7 days before infection (study day 0) and continuing for 28 days postinfection. On day 0, horses were stressed by transport and challenged with 1 million S. neurona sporocysts per horse. Sequential neurologic examinations were performed, and serum and cerebrospinal fluid were collected and assayed for antibodies to S. neurona. All horses in the control group developed neurologic signs, whereas only 71 and 40% of horses in the 2.5 and 5.0 mg/kg ponazuril groups, respectively, developed neurologic abnormalities. This was significant at P = 0.034 by using Fisher exact test. In addition, seroconversion was decreased in the 5.0 mg/kg group compared with the control horses (100 vs. 40%; P = 0.028). Horses with neurologic signs were killed, and a post-mortem examination was performed. Mild-to-moderate, multifocal signs of neuroinflammation were observed. These results confirm that treatment with ponazuril at 5.0 mg/kg minimizes, but does not eliminate, infection and clinical signs of EPM in horses.

Prevalence of antibodies against Toxoplasma gondii was studied in 534 pregnant women and 40 domestic cats in Grenada, West Indies. Antibodies (IgG) for T. gondii were sought in human sera by an enzyme-linked immunosorbent assay and in cat sera by using the modified agglutination test (MAT). Antibodies were found in 57 % of pregnant women. Seroprevalence increased with age; 51% of 15- to 19-yr-old women (100 total) had antibodies versus 60% of 20- to 24-yr-old women (127 total). Antibodies to T. gondii (MAT, 1:25 serum dilution) were found in 35% of cats; titers were 1:25 in 7 cats, 1:50 in 4 cats, and 1:500 in 3 cats. Epidemiological data suggested that the ingestion of food or water contaminated with oocysts was an important mode of transmission of T. gondii to women.

A cattle-drinking pool in nature reserve “Zwin” on the Belgian coast contained free-living third-stage infective filaroid juveniles. These juveniles clearly differ morphologically from all known nematodes. Morphological and molecular analyses indicate a position within the Filaroidea. The aberrant biology of this nematode, namely, a free-living stage in an aquatic environment, is unknown within this superfamily, and the evolution of the parasitic phenotype to a free-living state is generally thought to be unlikely. However, the obtained placement in the small subunit molecular phylogenetic tree suggests that this free-living stage is most likely a secondary adaptation. It is reasonable to assert that nematodes with complex life cycles still have the genetic potential for a reversion from parasitism to a (partial) free-living stage.

Antibodies to Toxoplasma gondii were assayed in sera of 266 humans from 71 farms located at Rondônia State, Western Amazon, Brazil, by the modified agglutination test (MAT) and the indirect immunofluorescent antibody test (IFAT). Antibodies were found in 195 humans (73.3%), with MAT titers of 1:25 in 11, 1:50 in 11, 1:100 in 16, 1:200 in 27, 1:400 in 38, 1:800 in 37, 1:1,600 in 22, and 1:3,200 or higher in 33. From the 71 farms visited, 69 had seropositive humans. Prevalence of anti-T. gondii antibodies increased with age of the people (P < 0.05), and no difference was observed in the occurrence by gender (P > 0.05). A sanitary questionnaire was applied in each farm, and statistical association between the serologic status and several variables were analyzed. Home-grown vegetable consumption and origin of drinking water (well or river) were the independent variables that displayed significant association (P = 0.002 and 0.02, respectively). Higher values of occurrence were found in people with consumption of home-grown vegetables (76.1%) and people that drink well water (75.4%) compared with people that did not consume this type of food (61.9%) and drink river water (55.2%). By IFAT (≥1:16), 194 of 266 (73%) humans were seropositive and there was a good correlation between MAT and IFAT.

Humidity and temperature have been considered important factors affecting the infectivity of Fasciola hepatica to its molluscan host. One hundred and thirty laboratory-reared Lymnaea humilis were exposed for 4 hr to the miracidia of F. hepatica over a pH range from 4.0 to 10.0, and their rates of survival were compared with 130 similarly treated but unexposed control snails. All control snails died within 24 hr at pH 4.0, but they showed better survival at pH 5.0–10.0. Their sensitivity to solutions with high and low pH, however, was increased if kept in the presence of F. hepatica miracidia. Snails exposed at pH 5.0 died within 24 hr, whereas most other pHs also affected survival such that by day 18 only those snails exposed at pH 7.2 remained alive. The increased sensitivity of the snails to pH could be explained by a damage-mediated release of parasite enzymes, because infectivity was highest at pHs associated with the lowest host mortality.

Serum was collected from laboratory-reared Virginia opossums (Didelphis virginiana) to determine whether experimentally infected opossums shedding Sarcocystis neurona sporocysts develop serum antibodies to S. neurona merozoite antigens. Three opossums were fed muscles from nine-banded armadillos (Dasypus novemcinctus), and 5 were fed muscles from striped skunks (Mephitis mephitis). Serum was also collected from 26 automobile-killed opossums to determine whether antibodies to S. neurona were present in these opossums. Serum was analyzed using the S. neurona direct agglutination test (SAT). The SAT was modified for use with a filter paper collection system. Antibodies to S. neurona were not detected in any of the serum samples from opossums, indicating that infection in the opossum is localized in the small intestine. Antibodies to S. neurona were detected in filter-paper– processed serum samples from 2 armadillos naturally infected with S. neurona.

Experimental murine cysticercosis caused by Taenia crassiceps has proved to be a useful model with which to test the efficacy of new vaccine candidates and delivery systems against pig cysticercosis. A high level of protection against murine cysticercosis was previously observed by intramuscular or intradermal DNA immunization with the use of the sequence of the recombinant KETc7 antigen cloned in pcDNA3 (pTc-sp7). To determine the effect of KETc7 differential expression in DNA vaccination, KETc7 was cloned in pGEM 11Zf( ) under the control of the tissue-specific regulatory promoter phosphoenolpyruvate carboxykinase (pPc-sp7). A high level of protection was induced by intrahepatic immunization with pPc-sp7, pTc-sp7 and the empty vector in the absence of any specific immunity. The empty vector pGEM 11Zf( ), the plasmid with the highest content of CpG sequences, provided to the most efficient protection. This protection was related to an increased number of splenocytes, enhanced nonspecific splenocyte proliferation, and intensified intrahepatic INF-γ production. Overall, intrahepatic plasmid CpG-DNA immunization provokes an exacerbated nonspecific immune response that can effectively control Taenia crassiceps cysticercosis.

Acute toxoplasmosis was diagnosed in 3 gray squirrels (Sciurus carolensis) from Louisiana and Pennsylvania. The predominant lesion was multifocal necrosis in several organs, especially of the lymph nodes. Numerous Toxoplasma gondii tachyzoites were seen in lesions, and the diagnosis was confirmed immunohistochemically by reaction with polyclonal T. gondii-specific antibodies. Tissue cysts were seen in several organs, including lung alveoli. The presence of tissue cysts in alveoli of pet squirrels maybe of public health concern if tissue cysts excreted in nasal secretions are swallowed by children.

Annual spring relapse of blood parasite infections in birds is believed to be the result of hormonal changes associated with breeding. As part of a larger study on the epizootiology of Haemoproteus danilewskyi in blue jays in south-central Florida, we studied the relationship between H. danilewskyi infections and levels of luteinizing hormone, prolactin, progesterone, testosterone, estradiol, and corticosterone. We found a positive association between intensity of H. danilewskyi infection and corticosterone levels in females but not in males. We also found no association between infection and levels of prolactin, luteinizing hormone, progesterone, testosterone, or estradiol in males or females. In addition, we found a positive relationship between levels of corticosterone and handling time and between corticosterone and testosterone levels. We suggest a possible influence of corticosterone on spring relapse of Haemoproteus spp. infections in birds but provide no support for the influence of breeding hormones on relapse of these parasites.

A study was conducted to determine the prevalence and geographical distribution of Sarcocystis spp. infecting 2 subspecies of caribou (Rangifer tarandus) inhabiting Newfoundland and Labrador and its potential definitive hosts. Muscle samples of caribou were obtained, primarily from hunters, and feces of red foxes (Vulpes vulpes) and wolves (Canis lupus), from trappers, and Husky dogs (Canis familiaris), from owners. Histological sections of muscle and flotation methods for feces were used for parasitic detection. Sarcocystis sp. infected more than 50% of barren-ground caribou (R. t. tarandus) from 4 locations in Newfoundland, but it was significantly greater in the north, where 99% of woodland caribou (R. t. caribou) from Labrador harbored the infection. Sporocysts were observed in 27 of 32 red foxes from eastern and northern Newfoundland, whereas 15 of 15 wolves and 22 of the 38 Husky dogs were infected. Wolves and red foxes probably acquired the infection through scavenging, and Husky dogs, from meat they were fed.

The prevalence of agglutinating antibodies to Toxoplasma gondii was examined in striped skunks (Mephitis mephitis), opossums (Didelphis virginiana), and raccoons (Procyon lotor) from 8 cities in Connecticut. Ten (42%) of the 24 striped skunks, 2 of 7 (29%) opossums, and 12 of 12 (100%) raccoons were positive at dilutions of 1:50 or greater. These results suggest that T. gondii is prevalent in the environment, or prey items, or both, of these omnivores in Connecticut.

Gnathiid isopods are common ectoparasites of fish on the Great Barrier Reef, Australia. While screening for appropriate markers for phylogenetic studies of gnathiids, we found that primers for 12S and 16S rDNA preferentially amplified the host fish DNA instead of gnathiid DNA. This amplification occurred even when using gnathiids that were not engorged with host blood and adult gnathiids that do not feed on fish blood. This method could be used in host–parasite studies to identify hosts without having to sample parasites directly from the host (which can be costly and requires considerable skill in a marine environment). Target ribosomal DNA sequences can be amplified from total DNA extracted from parasites that are captured in funnel traps or plankton tows. Sequence data from these can be used to identify the hosts that gnathiids were feeding on before capture.

Molecular analysis of parasite genomes will require new molecular genetic tools. The nat1 gene of Streptomyces noursei encodes nourseothricin acetyltransferase, conferring resistance to the aminoglycoside antibiotic nourseothricin. Electroporation of nat1 cassettes into RH or Prugniaud strains of Toxoplasma gondii allows for selection of stable nourseothricin-resistant clones.

Urotocus rossittensis occurs in the bursa Fabricii of the scarlet-rumped tanager, Ramphocelus passerinii, and the common bush tanager, Chlorospingus ophthalmicus, from the Area de Conservación Guanacaste, Costa Rica. Morphological examination of type material of U. fusiformis and U. kenyensis and Costa Rican specimens suggests that U. fusiformis and U. kenyensis are indistinguishable from U. rossittensis. Confirmed accounts of Urotocus spp. refer to a single adult morphotype whose geographic distribution includes the Palearctic, Africa, Nearctic, and northern Neotropics.

The neutral theory of biodiversity and biogeography is a null model of community structure that suggests that it may be possible to explain the richness and relative abundance of species through neutral processes of immigration, extinction, and speciation, without resort to interactive processes such as competition. There have been no attempts to fit neutral models to parasite communities to date. The nature of parasite communities, however, challenges the basic assumptions of neutral theory. In particular, the spatially dynamic relationships between hosts as habitat patches result in immigration rates that are in a constant state of flux. In addition, the partial compositional overlap of many component communities means that they can affect each other's process rates, which violates the zero-sum assumptions of neutral theory. Despite these obstacles, many of the patterns that neutral theory seeks to explain are still present in parasite communities. Far from being an esoteric special case, parasite communities are ubiquitous in nature and, therefore, any attempts to produce unified theoretical frameworks should accommodate the characteristics of parasite communities, or risk obsolescence.