Members of the Anisakidae are known to infect over 200 pelagic fish species and have been frequently used as biological tags to identify fish populations. Despite information on the global distribution of Anisakis species, there is little information on the genetic diversity and population structure of this genus, which could be useful in assessing the stock structure of their fish hosts. From 2005 through 2008, 148 larval anisakids were recovered from Pacific sardine (Sardinops sagax) in the California Current upwelling zone and were genetically sequenced. Sardines were captured off Vancouver Island, British Columbia in the north to San Diego, California in the south. Three species, Anisakis pegreffii, Anisakis simplex ‘C’, and Anisakis simplex s.s., were identified with the use of sequences from the internal transcribed spacers (ITS1 and ITS2) and the 5.8s subunit of the nuclear ribosomal DNA. The degree of nematode population structure was assessed with the use of the cytochrome c oxidase 2 (cox2) mitochondrial DNA gene. All 3 Anisakis species were distributed throughout the study region from 32°N to 50°N latitude. There was no association between sardine length and either nematode infection intensity or Anisakis species recovered. Larval Anisakis species and mitochondrial haplotype distributions from both parsimony networks and analyses of molecular variance revealed a panmictic distribution of these parasites, which infect sardines throughout the California Current ecosystem. Panmictic distribution of the larval Anisakis spp. populations may be a result of the presumed migratory pathways of the intermediate host (the Pacific sardine), moving into the northern portion of the California Current in summer and returning to the southern portion to overwinter and spawn in spring. However, the wider geographic range of paratenic (large piscine predators), and final hosts (cetaceans) can also explain the observed distribution pattern. As a result, the recovery of 3 Anisakis species and a panmictic distribution of their haplotypes could not be used to confirm or deny the presence of population subdivision of Pacific sardines in the California Current system.

Populations of North American waterbirds, particularly lesser scaup, have been declining due to habitat disturbance, changing food resources, contaminants, bad water quality, and competition. However, epizootic diseases, including parasitism, may also play an important role in further decline. Trematode-associated mortality of migrating waterbirds, mainly American coot and lesser scaup, has been occurring in the Upper Mississippi River National Wildlife and Fish Refuge since 2002. We examined the levels of infective stages of Cyathocotyle bushiensis and Sphaeridiotrema globulus in the invasive, intermediate host snail, Bithynia tentaculata, during the fall of 2005 and compared these to infection levels in moribund or dead bird hosts. Our results show different infection levels of these 2 parasites in the 2 bird species; C. bushiensis is found more frequently in coot, and S. globulus is more common in scaup. This result is interesting because both bird species are presumed to forage on the same snail population and thus should be experiencing the same extent of exposure. These differences in infections could be attributed to differences in resources of gastrointestinal tracts of coot and scaup, or host resistance. Alternatively, differences in feeding behaviors of coot and scaup may also contribute to differential infections of the 2 trematodes.

The nematode Skrjabinoptera phrynosoma is a stomach parasite of horned lizards in the genus Phrynosoma. This nematode demonstrates a distinctive life cycle wherein entire gravid females harboring infective eggs exit with lizard feces. Pogonomyrmex spp. harvester ants collect these females and feed them to their larvae, which are the only stages of the intermediate host that can become infected. We hypothesized that the seasonal dynamics of nematode abundance within lizard hosts would be correlated with the seasonal availability of suitable intermediate hosts. To describe seasonal variation of nematode population variables and elucidate the timing of critical events in the parasite life cycle, nematodes were collected from both hosts across three collection periods in the ant-and-lizard activity season of 2008 in the Alvord Basin of southeastern Oregon. Among 3 collection periods, and across the activity season, nematodes were harvested from individual Phrynosoma platyrhinos, and the distribution of developmental categories and body lengths of nematodes was analyzed to determine the seasonal change in nematode population composition. Pogonomyrmex spp. ants were collected in pit-fall traps and dissected to determine infection prevalence. The abundance of non-gravid female and juvenile nematodes collected from lizards' stomachs decreased significantly between the early and late collection period, which was likely a consequence of the sequential conversion of these developmental categories to gravid females. The presence of gravid female nematodes peaked in cloacal and fecal collections during mid-season. The body lengths of male nematodes increased as the activity season progressed, perhaps due to growth, but their abundance remained the same. Smaller juvenile nematodes were present in late-season collections from lizards, possibly indicating new acquisitions from infected ants. We propose that once a set population of male nematodes establishes in lizards' stomachs, newly acquired juvenile nematodes develop into non-gravid females that mate, become gravid females, and exit the lizard mid-season. We additionally suggest that the exit of females may be timed with the peak foraging activity of ant intermediate hosts and access to larval ants in the nests. Infection prevalence in the intermediate host was low, with only 1 of 6,000 dissected harvester ants containing a single larval nematode. The temporal dynamics of S. phrynosoma populations within P. platyrhinos at this northern locale is most likely driven by the seasonal availability of harvester ant intermediate hosts.

Seasonal occurrence of parasitic copepods in wild bullseye puffer, Sphoeroides annulatus (Pisces: Tetraodontidae), was analyzed in conjunction with variation of biotic and abiotic factors. Eleven samples were taken between February 2007 and February 2008 in Santa María La Reforma lagoon (northwestern coast of México). In total, 337 fish was examined; 5 parasitic copepod species were observed, including Acantholochus zairae, Caligus serratus, Lepeophtheirus simplex, Pseudochondracanthus diceraus, and Parabrachiella sp. The most common species were L. simplex, P. diceraus, and C. serratus (overall prevalence, 59, 53, and 35%, respectively), which significantly varied in prevalence and mean intensity between sampling months. A seasonal pattern was only observed for L. simplex, with higher infection levels in the warmest month than in the coldest month. Statistical analyses indicated that the intensity of L. simplex was positively correlated with water temperature. There were no significant differences in prevalence and intensity of infection among female and male hosts. At the component community level, species richness ranged between 4 and 5 during most of the study period, and no seasonality was observed in the number of individuals, Shannon diversity index, evenness index, or the Berger–Parker dominance index. At the infracommunity level, 4 descriptors used (mean species richness, mean number of individuals, mean Brillouin's diversity index, and mean Berger–Parker index) varied significantly between sampling months, but no seasonality was observed, except for a slight increase in the number of individuals during the warmest month. A significant positive association was detected between number of individuals and water temperature and between host size and both species richness and number of individuals. This is the first account of the ecology of these 5 parasitic copepods. Although no significant association was detected between fish condition factor and the burden of parasitic copepods, given the high occurrence of the caligid copepod L. simplex, we suggest that this copepod could represent a threat for the culture of S. annulatus.

A collection of laelapine mites from small mammals in the Galapagos Islands are identified and their host distributions reviewed. Two species of native rodents, Aegialomys galapagoensis and Nesoryzomys narboroughii, were infested only with laelapine species typical of Neotropical oryzomyine rodents; Rattus rattus was infested with Laelaps nuttalli, a host-specific ectoparasite endemic to Old World Rattus. A synopsis of Gigantolaelaps Fonseca is provided and we describe a new laelapine mite, Gigantolaelaps aegialomys n. sp., from the pelage of the rodent A. galapagoensis on Santa Fe Island. The new species has strong morphological affinities with a subgroup of Gigantolaelaps associated with a group of semiaquatic oryzomyine rodents (Holochilus, Nectomys, Sooretamys, Pseudoryzomys, Oryzomys palustris). The other nominal species of this group, Gigantolaelaps mattogrossensis (Fonseca, 1935) and Gigantolaelaps goyanensis Fonseca, 1939, are characterized by 10 setae on Tibia IV, large metapodal shields, and spiniform setae on Coxae I. Gigantolaelaps aegialomys is distinguished from these species by a lack of clearly spiniform setae on Coxa I, with setiform distal seta longer than the proximal; metapodal shields about the same size as the stigma; less than 100 µm separating the first pair of sternal setae.

Amyloodinium ocellatum is a parasitic dinoflagellate that infects warm-water marine and estuarine fishes and causes mortalities in aquaculture. Its life cycle consists of 3 stages: a feeding trophont that parasitizes the gills and skin where it interferes with gas exchange, osmoregulation, and tissue integrity; a detached reproductive tomont; and a free-swimming infective dinospore. We compared the susceptibility and tolerance of juvenile spotted seatrout, Cynoscion nebulosus, and red snapper, Lutjanus campechanus, to this parasite by individually exposing fish in 3-L aquaria (at 25 C and 33 practical salinity units) to several dinospore doses over different time periods and quantified the size and number of resulting trophonts. We estimated the trophont detachment rate and trophont size at detachment, the 24-hr dinospore infection rate, the dinospore 48-hr median lethal dose (LD₅₀), and the trophont lethal load at the 48-hr LD₅₀. There were no significant differences in dinospore infection rates or dinospore lethal doses between spotted seatrout and red snapper; however, trophonts remained attached longer and attained a larger size in red snapper than in spotted seatrout. The trophont lethal load was significantly higher in spotted seatrout than in red snapper. A proposed model simulating the trophont dynamics reflected our experimental findings and showed that A. ocellatum reproductive success is linked both to the number of dinospores and the size of the trophont, factors that, in turn, are linked to the time the trophont spends on the host and the number of trophonts the host can tolerate.

Three new species of Demidospermus Suriano, 1983 were found in mixed infections on the gills of 4 pimelodid catfish collected in the Amazon River basin around Iquitos, Peru, namely, Demidospermus peruvianus n. sp. from Pimelodus ornatus Kner, Pimelodus blochii Valenciennes, Pimelodus sp. (type host), and an unidentified pimelodid. Demidospermus peruvianus n. sp. (type host, Pimelodus sp.) is characterized by the presence of the male copulatory organ with 2 lateral, thickened projections, and the vagina with a loop at its middle. Demidospermus curvovaginatus n. sp. from P. ornatus, Pimelodus sp. (type host), and Pimelodidae gen. sp. differs from its congeners by the presence of a funnel-shaped vagina directed posteriorly and a male copulatory organ with a small lateral projection at its base. Demidospermus striatus n. sp. from P. blochii and Pimelodus sp. (type host) is distinguished by a cup-shaped vagina with conspicuous ridges at the perimeter of the aperture with a short tube directed anteriorly. In addition, Urocleidoides lebedevi Kritsky and Thatcher, 1976, a parasite originally described from the gills of Pimelodus grosskopfii (type host) from Colombia, is transferred to Demidospermus, based on morphological characters that we observed in type specimens.

A description and illustrations are given for Myrsidea povedai n. sp. from the black-and-yellow silky-flycatcher Phainoptila melanoxantha. The female of M. povedai is distinguished from those of other species of Myrsidea from Costa Rican passerine hosts by a unique combination of the following characteristics: (1) well-developed hypopharynx, (2) well-defined median gap in the rows of tergal setae and another gap between groups of setae on lateral side of tergites II–VII and the most central seta, and (3) enlarged metanotum with at least 23 setae. These characters place M. povedai close to Myrsidea campestris from Euneornis campestris and Myrsidea marini from Pezopetes capitalis (both from the Emberizidae). The female of M. povedai can be easily separeted from both aforementioned species by the abdomen without conspicuously enlarged tergites. Moreover, the male of M. povedai is characterized by a unique male genital sclerite, which is quite long (0.13–0.15), tapered apically, with a long median line, and without subapical processes. This is the first record of a chewing louse from this host and the first record of Myrsidea from the passerine family Bombycillidae. All 7 birds examined in Costa Rica in 2010 were parasitized with M. povedai. Mean abundance was 11.6, with intensity range 4–27 lice per bird.

Dicyemids (phylum Dicyemida) are endoparasites, or endosymbionts, typically found in the renal sac of benthic cephalopod molluscs. The body organization of dicyemids is very simple, consisting of only 9 to 41 somatic cells. Dicyemids appear to have no differentiated tissues. Although categorization of somatic cells, to some types, is based on differences in the pattern of cilia and their position in the body, whether or not these cells are functionally different remains to be revealed. To provide insight into the functional differentiation, we performed whole mount in situ hybridization (WISH) to detect expression patterns of 16 genes, i.e., aquaglyceroporin, F-actin capping protein, aspartate aminotransferase, cathepsin-L-like cysteine peptidase, Ets domain-containing protein, glucose transporter, glucose-6-phosphate 1-dehydrogenase, glycine transporter, Hsp 70, Hsp 90, isocitrate dehydrogenase subunit alpha, Rad18, serine hydroxymethyltransferase, succinate-CoA ligase, valosin-containing protein, and 14-3-3 protein. In certain genes, regional specific expression patterns were observed among somatic cells of vermiform stages and infusoriform larvae of dicyemids. The WISH analyses also revealed that the Ets domain-containing protein and Rad18 are molecular markers for agametes.

Understanding the genetic variation within a parasitic species is crucial to implementing successful control programs and preventing the dispersal of drug resistance alleles. We examined the population genetics and structure of canine heartworm (Dirofilaria immitis) by developing a panel of 11 polymorphic microsatellite loci for this abundant parasite. In total, 192 individual nematodes were opportunistically sampled from 9 geographic regions in the United States and Mexico and genotyped. Population genetic analyses indicate the presence of 4 genetic clusters. The canine heartworm samples used in this study were characterized by low heterozygosity, with eastern and central North America experiencing high levels of reciprocal gene flow. Geographic barriers impede the movement of vectors and infected hosts west of the Rocky Mountains and south of the Central Mexican Plateau. This, combined with corridors of contiguous habitat, could influence the spread of drug resistance alleles.

Extracted DNA from 28 Histomonas meleagridis-infected avian tissue samples from multiple hosts and geographic locations was analyzed for variation in the 5.8S rRNA and the flanking internal transcribed spacer regions (ITS 1 and ITS 2). Samples were amplified by polymerase chain reaction, sequenced, and compared with known sequences from GenBank accessions of H. meleagridis and other related protozoa. The analyses revealed significant genetic variation within H. meleagridis sequences and suggested the possibility of multiple genotypes within the samples or a possible misdiagnosis. Related protozoa found in some samples were mostly identified as Tetratrichomonas spp. However, 1 sample had a 93% identity to Simplicimonas similis, a newly described organism, suggesting the possibility of a new pathogen in poultry. A phylogenetic tree analyzing the 5.8S and flanking ITS regions was inconclusive and we were unable to resolve all H. meleagridis into a single grouping. In contrast, a tree constructed only on the 5.8S rRNA grouped all but 1 H. meleagridis sample into 1 clade, including GenBank accessions submitted from Europe. This suggests that the 5.8S region alone is more reliable in identifying H. meleagridis than are the combined 5.8S and flanking ITS regions. There was no correlation between genotypes and host species or geographic location, suggesting that H. meleagridis moves freely between multiple avian species in the sampled regions.

Little is known concerning the prevalence of Toxoplasma gondii infection in pigs in Mexico. Accordingly, antibodies to T. gondii were determined in 1,074 domestic pigs in Durango, Mexico using the modified agglutination test. Two groups (A, B) of pigs were sampled: Group A pigs (n  =  555) were raised in 3 geographical regions in Durango State and Group B pigs (n  =  519) were from Sonora State but slaughtered in Durango City. Overall, antibodies to T. gondii were found in 136 (12.7%) of 1,074 pigs with titers of 1∶25 in 29, 1∶50 in 23, 1∶100 in 18, 1∶200 in 22, 1∶400 in 12, 1∶800 in 8, 1∶1,600 in 2, and 1∶3,200 or higher in 22. Of the pigs raised in Durango State, seroprevalence varied with age, management, and the geographic region; pigs raised in backyards in the mountainous region had a significantly higher seroprevalence (32.1%) than those raised in the valley (13.0%) and the semi-desert regions (14.0%). In Group A pigs from Durango, seroprevalence of T. gondii infection was significantly higher in pigs older than 8 mo (19.5%) than in younger pigs (10.9%). In the whole pig population (Groups A and B together), seroprevalence was higher in pigs raised in Durango (16.0%) than in those raised in Sonora (9.1%) and higher in mixed-breed pigs (15.7%) than in pure-bred pigs (10.3%). This is the first, in-depth study on the seroprevalence of T. gondii infection of pigs in Mexico and the first report on pigs from Durango State, Mexico. Results indicate that infected pork is likely an important source of T. gondii infection for humans in Durango State.

Toxoplasma gondii is an important zoonotic parasite with a worldwide distribution. It infects about one-third of the world's population, causing serious illness in immunosuppressed individuals, fetuses, and infants. Toxoplasma gondii biology within the host cell includes several important phases: (1) active invasion and establishment of a nonfusogenic parasitophorous vacuole in the host cell, (2) extensive modification of the parasitophorous vacuolar membrane for nutrient acquisition, (3) intracellular proliferation by endodyogeny, (4) egress and invasion of new host cells, and (5) stage conversion from tachyzoite to bradyzoite and establishment of chronic infection. During these processes, T. gondii regulates the host cell by modulating morphological, physiological, immunological, genetic, and cellular biological aspects of the host cell. Overall, the infection/development predispositions of T. gondii–host cell interactions overtakes the infection resistance aspects. Upon invasion and development, host cells are modulated to keep a delicate balance between facilitating and eliminating the infection.

In total, 462 tadpoles and salamander larvae of 8 species were examined for the presence of Gyrinicola batrachiensis from 5 locations in Nebraska. Infection by G. batrachiensis occurred in tadpoles of Rana blairi, Rana catesbeiana, Rana pipiens, and Bufo woodhousii. Tadpoles of Hyla chrysoscelis, Spea bombifrons, and Pseudacris maculata and larvae of Ambystoma mavortium were not infected with G. batrachiensis. Population structure, defined as prevalence, mean abundance, and mean intensity of G. batrachiensis, varied among tadpoles of different amphibian species and was determined by collection locality, developmental period of tadpole hosts, amphibian species co-occurrence, and different reproductive strategies of G. batrachiensis, or a combination. Gyrinicola batrachiensis observed in all ranid tadpoles and B. woodhousii tadpoles from where bufonids were the only anuran species present, confirmed to the didelphic haplodiploidy and monodelphic parthenogenetic reproductive strategies, respectively. However, tadpoles of B. woodhousii that co-occurred with tadpoles of R. pipiens at Cedar Creek were inconsistent with these predictions and contained both male and didelphic female nematodes, but at a low mean intensity (1.61 ± 0.70). Didelphic female nematodes from B. woodhousii tadpoles at Cedar Creek only produced thick-shelled eggs, whereas nematodes in R. pipiens tadpoles had a high mean intensity (14.88 ± 23.83) from this location and contained both thick-shelled and thin-shelled eggs in their respective uteri. More importantly, adult female nematodes from tadpoles of R. pipiens and B. woodhousii from Cedar Creek were morphologically more similar to each other than to female nematodes recovered from tadpoles of other anuran species, other locations, or both. These data suggest that when strains of G. batrachiensis are shared by tadpoles of different amphibian species that differ in developmental period, the nematodes have an intermediate reproductive strategy in amphibian species, with tadpoles having short development.

A survey of the helminth fauna of the blue-footed booby, Sula nebouxii, on Isla Isabel, off the Pacific coast of México, is presented. Eight parasite species were found: 4 digeneans (Galactosomum puffini, Mesostephanus microbursa, Opisthometra planicollis, and Renicola thapari), 3 nematodes (Contracaecum sp., Porrocaecum sp., and Tetrameres sp.), and 1 cestode (Tetrabothrius sp). All these species are reported for the first time in the blue-footed booby, and they represent 8 new locality records as well. Species accumulation curves suggest these 8 parasite species comprise the total helminth fauna of this population of boobies. The most frequent and abundant parasite was R. thapari, inhabiting the kidney of its host. In male boobies, a significant negative correlation was found between abundance of R. thapari and host body condition.

A specific fatty acid binding protein was isolated from Giardia lamblia, using an affinity column with butyric acid acting as a ligand in place of stearic acid. This method has proved to be more efficient than the one previously described using stearic acid as ligand. The purified fraction showed 8 electrophoretic bands of proteins, with molecular weights ranging between 8 and 80 kDa. This pattern is a consequence of the aggregation of a protein with a molecular weight of 8,215 Da, corresponding to the lower molecular weight band, the only one capable of binding to fatty acids. The labeled oleic acid bound to these purified proteins was replaced by a 100-fold greater concentration of taurocholate, glycocholate, deoxycholate, palmitic acid, and arachidonic acid, having a greater displacement of the bile salts than the free fatty acids.

Novel Hepatozoon spp. sequences collected from previously unrecognized vertebrate hosts in North America were compared with documented Hepatozoon 18S rRNA sequences in an effort to examine phylogenetic relationships between the different Hepatozoon organisms found cycling in nature. An approximately 500–base pair fragment of 18S rDNA common to Hepatozoon spp. and some other apicomplexans was amplified and sequenced from the tissues or blood of 16 vertebrate host species from the southern United States, including 1 opossum (Didelphis virginiana), 2 bobcats (Lynx rufus), 1 domestic cat (Felis catus), 3 coyotes (Canis latrans), 1 gray fox (Urocyon cinereoargenteus), 4 raccoons (Procyon lotor), 1 pet boa constrictor (Boa constrictor imperator), 1 swamp rabbit (Sylvilagus aquaticus), 1 cottontail rabbit (Sylvilagus floridanus), 4 woodrats (Neotoma fuscipes and Neotoma micropus), 3 white-footed mice (Peromyscus leucopus), 8 cotton rats (Sigmodon hispidus), 1 cotton mouse (Peromyscus gossypinus), 1 eastern grey squirrel (Sciurus carolinensis), and 1 woodchuck (Marmota monax). Phylogenetic analyses and comparison with sequences in the existing database revealed distinct groups of Hepatozoon spp., with clusters formed by sequences obtained from scavengers and carnivores (opossum, raccoons, canids, and felids) and those obtained from rodents. Surprisingly, Hepatozoon spp. sequences from wild rabbits were most closely related to sequences obtained from carnivores (97.2% identical), and the sequence from the boa constrictor was most closely related to the rodent cluster (97.4% identical). These data are consistent with recent work identifying prey–predator transmission cycles in Hepatozoon spp. and suggest this pattern may be more common than previously recognized.

Aplectana novaeguineae n. sp. (Ascaridida, Cosmocercidae) from the intestines of Sphenomorphus pratti (Squamata: Scincidae) is described and illustrated. Aplectana novaeguineae n. sp. represents the 48th species assigned to the genus and the 5th species of the genus reported from the Australo-Papuan region. It is most similar to Aplectana leesi, Aplectana linstowi, and Aplectana krausi in that females of only these 4 species exhibit a swollen anterior vulvar lip. These 4 species may be separated by spicule length and caudal papillae arrangement. Spicules of A. linstowi and A. krausi are less than 200 µm in length, A. leesi between 200 and 250 µm, and A novaeguineae greater than 400 µm. Aplectana linstowi possesses 18 post cloacal papillae, A. leesi and A. krausi each possess 10, and A. novaeguineae possesses 8. In addition, Paraleptonema ranae has been assigned to Aplectana as Aplectana fujianae nom. nov.

Two new allocreadiid trematodes are described from the intestine of characid fishes from Rio Itaya, a tributary of the Amazon River near Iquitos, Peru. Auriculostoma foliaceum n. sp. is described from Bryconops cf. caudomaculatus (Günther, 1864) Backup, 2003, and Auriculostoma diagonale n. sp. from Stethaprion cf. erythrops Cope, 1870. Auriculostoma foliaceum n. sp. differs from its congeners by having a foliated lateral margin and lacking vitelline follicles in the forebody. Auriculostoma diagonale n. sp. differs from all of its congeners except Auriculostoma platense (Szidat, 1954) Scholz, Aguirre-Macedo, and Choudhury, 2004, by having oblique rather than tandem testes and differs from A. platense by having a much more elongated cirrus sac. Phylogenetic hypotheses based on analyses of 3 alignments of partial sequences of the 28S rDNA gene conducted in this study provide a framework for revaluating the classification of Allocreadiidae Looss, 1902. The present analyses show that (1) Paracreptotrematina limi Amin and Myer, 1982, does not belong in the Allocreadiidae; (2) Creptotrematina aguirrepequenoi Jiménez-Guzmán, 1973 (spelled here without diacritic mark), does not belong to Paracreptotrematina Amin and Myer, 1982, but remains in the Allocreadiidae; (3) Auriculostoma astyanace Scholz, Aguirre-Macedo, and Choudhury, 2004, is closely related to C. aguirrepequenoi and Wallinia chavarriae Choudhury, Hartvigsen-Daverdin, and Brooks, 2002; (4) Megalogonia Surber, 1928, is retained as an accepted genus in the Allocreadiidae; and (5) Margotrema Lamothe-Argumedo, 1970, is found to be nested among 3 species of Crepidostomum Braun, 1900.

The Attwater's prairie chicken (APC; Tympanuchus cupido attwateri Bendire, 1894) has been a federally listed endangered species since 1967. Several captive propagation programs consisting of small populations are being used to keep this species from extinction. Fecal samples were collected from APCs in April 2007 and again in August 2008 from 2 separate captive propagation facilities in Texas after clinical signs of coccidiosis were observed. One Eimeria species was observed (Eimeria attwateri), which we describe as a putative new species. Sporulated oocysts are ellipsoidal, 30.0 × 18.4 (27.4–31.3 × 16.0–22.4) µm. Oocysts have a smooth wall 0.7 µm thick and lack both a micropyle and oocyst residuum, but 1 ellipsoidal polar granule is present, 2.3 × 1.9 (2.1–2.4 × 1.7–2.0) µm. Sporocysts have a nipple-like Stieda body with a rounded opposite end and are 14.0 × 7.1 (10.2–16.8 × 6.0–9.2) µm. The sporocysts contain a sporocyst residuum usually consisting of 2–4 dispersed globules, and each sporozoite contains 2 large posterior spheroid refractile bodies 3.4 µm wide. Nucleotide sequence amplified from the 18S rDNA does not match any DNA sequence information for publicly available Eimeria species, and phylogenetic reconstructions place this species with other eimerians from Galliformes. The discovery of a potentially pathogenic species of Eimeria in captive APCs is of great importance, and managers should be aware of the potential devastating effect(s) this parasite could have on the APC conservation programs.

A new oxyurid nematode Syphacia hodarae n. sp. is described from the cecum and rectum of the cricetid rodent Calomys laucha Fischer, 1814 (Sigmodontinae, Phyllotini), captured in an agroecosystem of central Argentina. The new species is distinguished from other members of the genus mainly by the shape of the cephalic plate, presence of cervical alae in females, absence of lateral alae, and absence of deirids. Some characters are shared with Syphacia carlitosi, a parasite of Akodon azarae from the wetlands in Argentina. However, S. hodarae can be differentiated from this species by the absence of ornamentation on the accessory hook of the gubernaculum, length of spicule and gubernaculum, size of the eggs, and distance to the vulva from the anterior end. This is the first record of a Syphacia species from the tribe Phyllotini in Argentina, and the first time a Syphacia species is reported from C. laucha.

Haemoproteus (Parahaemoproteus) micronuclearis n. sp., Haemoproteus (Parahaemoproteus) nucleofascialis n. sp., Haemoproteus (Parahaemoproteus) paranucleophilus n. sp., and Haemoproteus (Parahaemoproteus) homobelopolskyi n. sp. (Haemosporida, Haemoproteidae) are described from African passeriform birds based on the morphology of their blood stages and segments of the mitochondrial cytochrome b gene. Red-billed quelea (Quelea quelea), red-headed malimbe (Malimbus rubricollis), and black-headed weaver (Ploceus melanocephalus) are the type vertebrate hosts of new hemoproteids. It is probable that new species have wide distribution in weavers in sub-Saharan Africa. Both H. micronuclearis and H. nucleofascialis can be readily distinguished from other avian hemoproteids by tiny, compact microgametocyte nuclei that are significantly smaller than macrogametocyte nuclei and are a rare character of hemosporidian parasites. Gametocytes of H. paranucleophilus are closely appressed to the erythrocyte nuclei and do not touch the erythrocyte envelope along their entire margin at all stages of their development, including fully grown gametocytes. A particularly distinctive feature of H. homobelopolskyi development is the presence of circumnuclear dumbbell-shaped macrogametocytes. Illustrations of blood stages of the new species are given, and morphological and phylogenetic analyses identify the DNA lineages that are associated with these parasites. Numerous recent studies show that some lineages of hemoproteids are often present in birds belonging to different families. As a result, the use of the host family as a taxonomic character should be questioned and preferably discouraged in hemoproteid taxonomy, particularly with regard to the parasites of passerine birds. Microscopic identification of avian hemoproteids requires comparison of Haemoproteus species described from birds of different families, as is an established practice with avian Plasmodium spp. Development of bar-coding techniques remains essential in taxonomic and field studies of hemosporidian parasites.

Cucullanus dodsworthi Barreto, 1922 was originally described from the checkered puffer fish, Sphoeroides testudineus (Linnaeus), from Brazilian waters. New material of this nematode species was recovered from the same type host species from Mexican waters off the Yucatán Peninsula. This material was compared with Brazilian specimens. Although Mexican material closely resembles the original description of C. dodsworthi in general appearance, previously undescribed characters, as observed by light and scanning electron microscopy, are described for the first time in this species from both Brazilian and Mexican specimens. These characters include lateral body alae or conspicuous lateral fields that begin in the cervical region and end anterior to first pair of adcloacal papillae in males and at the anus level in females, cephalic and caudal alae absent; presence of pseudobuccal capsule with simple buccal frame well sclerotized with dorsal arrow structures, lateral structures, and lateral reniform structures; deirids, excretory pore, and postdeirids; slight anal protuberance in both sexes, unpaired precloacal papilla in males, phasmids near pair 10 in males and near tail tip in females; female with protruding vulvar lips and smooth eggs. In the absence of better descriptions of this genus, it can be concluded that C. dodsworthi is the only species of marine Cucullanus from the Americas that possesses lateral body alae. Molecular characterization of C. dodsworthi with SSU (18S) and ITS2 rDNA genes is included. A preliminary genetic comparison between SSU rDNA of C. dodsworthi, Truttaedacnitis truttae (Fabricius, 1794), and Dichelyne mexicanus Caspeta-Mandujano, Moravec and Salgado-Maldonado, 1999 places C. dodsworthi as a putative sister taxon to T. truttae. The finding of C. dodsworthi in Mexican marine waters also represents a new geographical record.

Paracreptotrema profundulusi n. sp. (Trematoda: Allocreadiidae) is described from the intestine of the freshwater fishes Profundulus punctatus and P. balsanus (Teleostei: Profundulidae) from the Tehuantepec and the Atoyac-Verde River basins, in Oaxaca, México, in the western extreme of Central America. The new species is distinguished from Paracreptotrema blancoi Choudhury, Pérez-Ponce de León, Brooks, and Daverdin, 2006, to which it is most similar, and from P. mendezi (Sogandares-Bernal, 1955) Choudhury, Pérez-Ponce de León, Brooks, and Daverdin, 2006, by the caeca extending beyond the testes, vitelline follicles that invade the postesticular area, and uterus with transverse loops located mainly between the testes and the genital pore. Paracreptotrema blancoi was collected from the same host species and also from Profundulus oaxacae. Here, we provide data that show its broad distribution in several river basins of Neotropical southern México, including the Papagayo River basin, Guerrero, México, and the Atoyac-Verde and Tehuantepec river basins, and other rivers in Oaxaca, México. Freshwater fishes of the Profundulidae are endemic to Central America and host a helminth fauna that includes at least 4 species found only in these hosts.

Sphaeromyxa cannolii sp. n. is described from the bile ducts of aquaria-maintained lined seahorses (Hippocampus erectus) from the Gulf of Mexico. Spores of the new species are linear, 17–18 µm long and 5–6 µm wide, with flattened tips; polar capsules measure 4 × 3 µm. Routine necropsies of H. erectus following planned death revealed liver inflammation, bile duct obstruction, bile accumulation, and myxozoan parasites in the bile ducts of 11 of 40 animals sampled (27.5%). The presence of S. cannolii in an aquaculture setting should prompt keepers to carefully quarantine new animals and exclude annelid fauna, a potential intermediate host of myxozoans.

Paraoistosomum novaeguineae n. gen., n. sp. is described based on specimens from the kidneys of a New Guinea crocodile Crocodylus novaeguineae collected in Papua New Guinea. The body shape and the topology of most internal organs of the new species are strongly reminiscent of Oistosomum caduceus Odhner 1902, the sole member of Oistosomum, a genus of unclear phylogenetic relationships and systematic position described from the Nile crocodile in Sudan in 1902 and never reported since then. At the same time, the new species has a number of significant differences from Oistosomum caduceus. Among them are much shorter intestinal ceca, a relatively larger ventral sucker, ovary anterolateral to ventral sucker (posterolateral in O. caduceus), vitellaria arranged in 2 clusters of loosely organized follicles at the level of ventral sucker (2 narrow long lateral fields in O. caduceus), much longer esophagus, and other characters. Most importantly, the new species has the genital pore situated at the anterior body end adjacent to the oral sucker, whereas O. caduceus has the genital pore in front of the ventral sucker. These dramatic differences suggest establishment of a new genus for the species from Papua New Guinea. The anatomy of P. novaeguineae n. gen., n. sp. suggests that these genera may not belong to the Plagiorchiidae, the current familial allocation of Oistosomum. Scarcity of material and lack of molecular data do not permit clarification of this problem at the present time.

With the use of 2 monoclonal antibodies (MAbs) against excretory/secretory (ES) antigens of adult Angiostrongylus cantonensis, a new method was developed for double antibody sandwich ELISA for the detection of circulating antigens (CAg). To evaluate the sensitivity of the new procedure, the CAg in sera of rats (80) and mice (15) infected with A. cantonensis, as well as CAg in sera of clinically confirmed angiostrongyliasis patients (70), were evaluated. Cross-reaction testing was used to determine the specificity of serum from patients infected with Ascaris lumbricoides, Trichinella spiralis, Toxoplasma gondii, Schistosoma japonicum, Paragonimus westermani, Clonorchis sinensis, Echinococcus granulosus, Spirometra, and Taenia solium, as well as normal healthy people. The results proved that the sensitivity and the specificity of the new method were totally effective for the detection of A. cantonensis CAg. The assay is highly sensitive, specific, and reproducible, with easy handling and excellent cost effectiveness, and thereby provides a new method for the accurate diagnosis of angiostrongyliasis.

Rapid and reliable detection and identification of coccidian oocysts are essential for animal health and foodborne disease outbreak investigations. Traditional microscopy and morphological techniques can identify large and unique oocysts, but they are often subjective and require parasitological expertise. The objective of this study was to develop a real-time quantitative PCR (qPCR) assay using melting curve analysis (MCA) to detect, differentiate, and identify DNA from coccidian species of animal health, zoonotic, and food safety concern. A universal coccidia primer cocktail was designed and employed to amplify DNA from Cryptosporidium parvum, Toxoplasma gondii, Cyclospora cayetanensis, and several species of Eimeria, Sarcocystis, and Isospora using qPCR with SYBR Green detection. MCA was performed following amplification, and melting temperatures (T_m) were determined for each species based on multiple replicates. A standard curve was constructed from DNA of serial dilutions of T. gondii oocysts to estimate assay sensitivity. The qPCR assay consistently detected DNA from as few as 10 T. gondii oocysts. T_m data analysis showed that C. cayetanensis, C. parvum, Cryptosporidium muris, T. gondii, Eimeria bovis, Eimeria acervulina, Isospora suis, and Sarcocystis cruzi could each be identified by unique melting curves and could be differentiated based on T_m. DNA of coccidian oocysts in fecal, food, or clinical diagnostic samples could be sensitively detected, reliably differentiated, and identified using qPCR with MCA. This assay may also be used to detect other life-cycle stages of coccidia in tissues, fluids, and other matrices. MCA studies on multiple isolates of each species will further validate the assay and support its application as a routine parasitology screening tool.

In June 2009, 27 guinea pigs kept at an animal petting facility at a zoo in Kanagawa Prefecture, Japan, were observed to scratch intensely, weaken, and develop lesions. Three sarcoptiform mites were found in skin scrapings taken from affected areas of 2 guinea pigs, and they were identified as Trixacarus caviae by morphological examination. This result confirmed the presence of T. caviae in Japan. For treatment, doses of 13.6–18.75 mg/head of selamectin were administered in a topical preparation applied to a single spot on the skin on the back of the neck, and no side effects were observed. In April 2010, a second outbreak of mange occurred at the zoo, and, following investigation, 2 mite eggs were observed. It was, therefore, thought probable that the mites had survived during the winter within nonclinical carriers. Accordingly, doses of 5.0–7.5 mg/head of selamectin were applied on days 0 and 28, after which clinical symptoms disappeared and general condition improved. This dose of selamectin was thus shown to be a suitable and economical treatment for guinea pigs infested with the mites. Because the mite is not always easily observed in infested guinea pigs and the potential for human infestation exists, clinicians should not hesitate to treat when the clinical presentation suggests infestation, particularly in a setting such as an animal petting facility, where large numbers of children and adults have direct contact with the animals.

Trichinella spp. infections of marine mammals pose a human health risk in Arctic regions where game meat is an important part of the diet. In the last decade, the grey seal population of the sub-arctic Baltic Sea has reached a level at which hunting is possible and seal meat is now available for food. We conducted a survey on the occurrence of Trichinella spp. in grey seals Halichoerus grypus (n  =  171) and ringed seals Phoca hispida botnica (n  =  56) of the Baltic Sea in the coastal waters of Finland, a highly Trichinella sp. endemic area. Muscle samples were examined by a mechanically assisted digestion method during 2006–2010. One grey seal was positive for Trichinella nativa, while all samples from ringed seals were negative. Even though just 1 grey seal was infected, the finding here emphasizes the importance of proper meat inspection of seals intended for human consumption, especially in areas with high infection pressure.

Between April 2008 and March 2009, we analyzed the pattern of coccidian oocysts present in the feces of the European bison (Bison bonasus L., 1758) and found 4 species (Eimeria bovis, E. canadensis, E. ellipsoidalis, E. zuernii) previously reported from this host and 3 species (Eimeria alabamensis, E. cylindrica, E. pellita) that are new host and locality records. All the species occurred in bison females, and only 4 occurred in males; E. bovis was the most prevalent in both sexes. The overall prevalence of Eimeria spp. invasion reached 34.7% in cows and 13.9% in bulls. The highest prevalence was noted in early spring, with a peak in April, and the lowest in late autumn and winter. The oocyst count per gram of feces (OPG) varied from 50 to 1,350; no symptoms of clinical coccidiosis were observed. We found a significant influence of winter aggregations of bison on shedding of coccidian oocysts. The prevalence and OPG values were higher in bison congregating in large numbers around winter-feeding sites in comparison to other sites. We suggest that the coming together of cows during the growing season impacts the gender-related differences in prevalence and the number of coccidian species involved. This observation probably results from an increased production of oocysts by sub-clinically infected individuals in high-density bison populations.

The protozoan Neospora caninum and the bacterium Brucella abortus are well-recognized causes of abortion in dairy cattle. Serum samples (n  =  240) from aborting (n  =  141) and at-risk (n  =  99) animals from 5 herds with high abortion rates in Punjab Province, Pakistan, were tested for antibodies to N. caninum using monoclonal antibody–based ELISA and for antibodies to B. abortus using the serum agglutination test. Antibodies to N. caninum and B. abortus were detected in 105 (43.8%) and 135 (56.3%) cattle, respectively. Prevalences of antibodies to N. caninum and B. abortus were higher in aborting cows (46.8% and 76.6%, P < 0.05) than in animals at risk (39.4% and 27.3%, P > 0.05). Sixty-six animals (27.5%) were seropositive to both N. caninum and B. abortus, and results showed no significant difference (P > 0.05) with respect to geographical district, breed, and age. This is the first report of N. caninum infection among dairy cattle herds in Pakistan.

Helisoma spp. snails are not susceptible to infection with miracidia of Schistosoma mansoni because the miracidia do not penetrate them. However, in view of the phylogenetic proximity and histocompatibility between Helisoma spp. and the normal intermediate host, Biomphalaria glabrata, schistosome miracidia conceivably could survive if experimentally introduced into the hemocoel of Helisoma spp. To test this hypothesis, schistosome-susceptible NIH albino B. glabrata, schistosome-resistant Salvador B. glabrata, and Helisoma duryi were injected with miracidia of S. mansoni, and the outcome was followed both by monitoring snails for infection for several weeks and by histological examination at 24 and 48 hr post-injection (PI). Patent infections developed in most NIH albino snails but in none of the Salvador B. glabrata or H. duryi individuals. Histological analysis showed a higher proportion of normal sporocysts in various tissues of NIH albino snails at both time periods relative to Salvador snails, which contained mostly sporocysts undergoing hemocytic encapsulation. In H. duryi, nearly all sporocysts were dead by 48 hr PI.

A simplified method for the culture of neonatal Biomphalaria glabrata snails on a diet of the cyanobacteria Nostoc sp. has been developed. Previous studies reported using Nostoc sp. grown on a mud-based medium to feed neonatal snails; this medium is difficult to prepare and gives inconsistent growth between laboratories due to variations in the soil used in its preparation. Here, we report the use of Bg-11 medium supplemented with sodium nitrate for the culture of Nostoc sp. as a food source for neonatal B. glabrata snails. Bacteria grown in the medium may be maintained in continuous culture and used exclusively as the nutrient source for neonatal snails. Neonatal snails fed with these bacteria exhibit a growth rate of 1–2 mm per week and may be transferred to standard culture conditions after 2–3 wk. Survival of neonatal snails after 2 wk on the Nostoc sp. diet was 60–80%, while survival after switching to standard culture conditions approached 100%. Analysis of the growth rate and survival of neonatal snails maintained on Nostoc sp., as well as the growth rate, survival, and sexual maturation of the resulting juvenile snails, showed that snails maintained using this method are phenotypically comparable to those maintained under other standard laboratory conditions.

Seroprevalence of Toxoplasma gondii infection in pigs in 10 regions of Zhejiang Province, China, was obtained by enzyme-linked immunosorbent assay (ELISA). Anti–T. gondii antibodies were found in 53.4% (434/813) of pigs. Results were analyzed by a chi-square (χ²) test. Differences were observed according to farm size, animal age, and sampling regions. Seroprevalences in pigs raised on small farms (71.4%) were significantly higher than that (42.7%) on large farms (P < 0.05), and seroprevalence increased progressively with age. The seroprevalence ranged from 28.1% to 66.0% in different regions, with Jiaxing having the lowest level (28.1%), followed by Hangzhou (36.0%) and Taizhou (42.0%). This is the first study on seroprevalence of T. gondii infection in pigs in Zhejiang Province.