Manifestations of infectious disease may represent host adaptations to avoid or reduce the effects of infection on host fitness, parasite manipulations that benefit the pathogen's fitness, or nonadaptive side effects of parasitism. Threespine stickleback fish (Gasterosteus aculeatus) from Alaska and the cestode macroparasite Schistocephalus solidus provide an excellent system for study of the effects of parasitism on host egg size because females in populations there are capable of producing clutches of eggs in the face of substantial infection, contrary to the inhibition of reproduction that has been observed in other stickleback populations or other species of fish. A side effect resulting in reduction of mean ovum mass among infected females was predicted based on the egg production process in female stickleback, the considerable energy and resource demands of S. solidus, and the chronic and progressive nature of the effects the macroparasite should have on the host fish. In each of 9 populations of G. aculeatus representing replicate natural experiments in lakes scattered across the Matanuska–Susitna Valley and the Kenai Peninsula of south-central Alaska and among all populations combined, the mean ovum mass of infected female fish is significantly reduced in comparison with that of uninfected females taken from the same population at the same time. Reduction in mean female egg mass ranged from 8 to 32% across all populations. To examine whether reduction in mean female ovum mass was a nonadaptive side effect or an adaptation, relatively large data sets from 2 of the populations were used. Mean ovum mass of infected females was predicted to decrease directly in relation to parasite index (PI) if the diminution in mean egg mass were the result of a nonadaptive side effect resulting from host nutrient loss. Alternatively, the absence of a relationship between PI and reduction in ovum mass is predicted if decreases in mean female ovum mass result from host or parasite adaptation (or both) because lightly infected hosts should show a response similar to that of heavily infected ones. In each of the 2 populations, there is a significant, negative relationship between mean female ovum mass and PI, demonstrating a correlation between the decrease in ovum mass and the level of infection. Thus, the results are consistent with the hypothesis that the reductions in mean female egg mass represent side effects of parasitism involving nutrient theft. Moreover, the proportional decline in egg mass with increasing PI apparently differed between the 2 populations, and there was no significant relationship between mean percent decrease in mean female ovum mass and mean PI across populations. These observations suggest that unknown ecological and evolutionary factors influence the degree of reduction in mean ovum mass in a population-specific manner.

Adult and mobile preadult sea lice Lepeophtheirus salmonis were incubated with mucus samples from rainbow trout (Oncorhynchus mykiss), coho salmon (O. kisutch), Atlantic salmon (Salmo salar), and winter flounder (Pseudopleuronectes americanus) to determine the response of L. salmonis to fish skin mucus as assessed by the release of proteases and alkaline phosphatase. There was variation in the release of respective enzymes by sea lice in response to different fish. As well, sea lice collected from British Columbia responded differently than New Brunswick sea lice to coho salmon mucus. Fish mucus and seawater samples were also analyzed using protease gel zymography to observe changes in the presence of low molecular weight (LMW) proteases after L. salmonis incubation. Significantly higher proportions of sea lice secreted multiple bands of L. salmonis–derived LMW proteases after incubation with rainbow trout or Atlantic salmon mucus in comparison with seawater, coho salmon, or winter flounder mucus. Susceptibility to L. salmonis infections may be related to the stimulation of LMW proteases from L. salmonis by fish mucus. The resistance of coho salmon to L. salmonis infection may be due to agents in their mucus that block the secretion of these LMW proteases or factors may exist in the mucus of susceptible species that stimulate their release.

Accelerated proliferation of the tick-borne bacterial pathogen Francisella tularensis was demonstrated in mice when the bacterium was injected together with salivary gland extract from Ixodes ricinus ticks. A significant increase in the numbers of bacteria was recorded in the dermal site of infection, the draining lymph nodes, and the spleen. Analysis of the expression of cytokine messenger ribonucleic acids showed polarization toward a Th2 profile. Salivary gland extract–mediated suppression of interleukin-12 and interferon-γ, the cytokines required for the expression of the protective immunity against tularemic infection, apparently contributed to the decreased resistance against this tick-transmitted pathogen.

Eastern Tubifex tubifex worms were exposed to Myxobolus cerebralis spores at 9, 13, 17, and 20 C in 1-L jars that contained sand, mud, or leaf litter as substrata. Beginning 60 days after exposure, water from each jar was filtered daily and examined for the presence of waterborne triactinomyxon spores (TAMs). On discovering a single TAM from an experimental jar, 48 T. tubifex worms from that jar were placed individually into 24-well plates. Spores released from individual infected T. tubifex worms were quantified to determine the first day of TAM release from infected worms, the infection rate, the total number of TAMs released per worm, and the duration of release. No TAMs were found in any of the jars incubated at 20 C or in uninfected, control worms at any temperature. The total number of TAMs released by infected worms in mud and sand was highest at 13 C compared with other temperatures. Infection rates among individual worms increased with temperature between 9 and 17 C. Higher temperatures (up to 17 C) induced earlier TAM releases among infected worms, and substratum did not influence this production parameter. The average duration of TAM release decreased as the temperature increased from 9 to 17 C, and there was a significant effect of substratum in the groups maintained at 13 and 17 C. In all temperature treatments between 9 and 17 C, the duration of release was least in the worms maintained in leaf litter, as was the total number of TAMs released during the experimental period and the median number of TAMs per production day.

Mesocestoides corti is a suitable model for studying cestode development because of its ability to reproduce asexually and segment in vitro. The cultured parasite is also capable of sexual differentiation and, probably, reproduction. To establish conditions that increase the efficiency of in vitro M. corti larvae (tetrathyridia) segmentation, we tested the effects of an inducing agent and some physical parameters in cultures. We found that a 5% CO₂–95% N₂ gas phase, an incubation temperature of 39 C (instead of 37 C), and a 24-hr pretreatment with trypsin (10⁵ BAEE/ml, BAEE = Nα-benzoil-l-arginine ethyl ester unit of trypsin activity) in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 20% fetal bovine serum (FBS) are able to increase individually or synergistically the segmentation rate of tetrathyridia. A segmentation rate of up to 100% was achieved on day 4 of culture, when all these conditions were used simultaneously, in comparison with an average rate of 40% obtained not before day 11 in cultures without any inducing treatment. Fetal bovine serum is essential for segmentation, and a concentration of 20% was established as the standard for induction.

Leishmania (Leishmania) amazonensis LV79 (MPRO/BR/72/M1841) has been adapted to grow at 33 C as amastigote-like (AL) organisms in modified UM-54 medium initially adjusted to a pH of 4.8–5.0. Axenic cultures could be routinely restarted from parasites recovered from footpad lesions obtained by inoculation of BALB/c mice with preadapted culture stages. Morphological features, proteinase activities, and infectivity of AL organisms were examined during the in vitro growth cycle, and differences were found between log- and stationary-phase parasites. Stationary-phase AL organisms were morphologically similar to lesion amastigotes, did not react with a paraflagellar rod-specific monoclonal antibody in western blots, and contained proteinase activities resolving identically to the enzymes of lesion amastigotes in gelatin gels. Whereas typical megasomes could be identified in about a third of the stationary-phase AL population, the organelles were rarely seen in log-phase organisms. Azocaseinolytic activity progressively increased during the exponential growth phase and reached its highest values (∼65–70% of those determined in lesion amastigotes) at the stationary phase; the association of total proteinase activity with increased expression of cysteine proteinases was indicated by the strong inhibition of azocasein hydrolysis by E-64, the intensified banding of the 28-, 31-, and 35-kDa proteinases in gelatin gels, and the higher susceptibility of stationary-phase AL organisms to l-leucine methyl ester. Although overall axenic amastigotes were less infective to BALB/c mice than were lesion-derived parasites, stationary-phase AL organisms were more infective than were log-phase parasites. Medium pH increased during the exponential growth phase, but dropped in the stationary phase, when the observed morphological, biochemical, and biological changes became apparent.

C57BL/6 mice were vaccinated with a bacterially expressed and purified polyhistidine-tagged full-length version of the microneme protein NcMIC3 (recNcMIC3) emulsified in Ribi Adjuvant System (RAS). Subsequently, they were challenged by intraperitoneal inoculation of 2 × 10⁶ live Neospora caninum tachyzoites. As controls, groups of mice received phosphate-buffered saline (PBS)-RAS alone (adjuvant control) or were treated with PBS before infection (infection control). The protective effect of vaccination was assessed by Neospora-specific polymerase chain reaction (PCR), immunohistochemical investigation of brain tissue, and serological means (enzyme-linked immunosorbent assay). Assessment by PCR performed on DNA from different organs revealed that in all treatment groups parasite DNA could only be detected in brain tissue. According to the PCR results, the recNcMIC3 vaccine conferred protection to 75% of mice (n = 16 in 2 independent experiments), whereas application of PBS-RAS and of PBS alone resulted in protection of 12.5% and 0% of mice, respectively (n = 16 as above). Mice in the PBS-treated infection control group were affected by evident clinical signs of neosporosis starting on day 6 postinfection (p.i.). Conversely, none of the animals treated with either PBS-RAS or recNcMIC3 exhibited any symptoms until day 21 p.i. Immunohistochemical staining of paraffin-embedded brain tissue sections confirmed the protective effect of recNcMIC3 vaccination. Quantitative Neospora-specific real-time PCR revealed that infection intensities were lower in the brain tissues of recNcMIC3-vaccinated mice compared with PBS-RAS–treated adjuvant control mice. Serological analysis showed that the protective effect observed in recNcMIC3-vaccinated mice was associated with a Th2-type IgG1 antibody response directed against native NcMIC3 and a mixed IgG1–IgG2a antibody response directed against the recombinant antigen itself. Taken together, these results demonstrated that recombinant NcMIC3 vaccine confers a significant protectivity against experimentally induced cerebral neosporosis in mice.

The effects of multiple infections on the host–parasite relationship between bluegill sunfish (Lepomis macrochirus) and parasitic glochidial larvae of the freshwater mussel Utterbackia imbecillis were examined. Naïve, young-of-the-year bluegills were infected with glochidia and placed in individual observation chambers. Each day, water was drained from each chamber and the numbers of dead glochidia, live glochidia, partially metamorphosed glochidia, and fully metamorphosed juvenile mussels were counted. The same fishes were infected a total of 4 times. After 2 infections, the fish began to exhibit evidence of acquired resistance to glochidia. During the third and fourth infections, this resistance was clearly evidenced by the marked increase in the percentage of dead and live glochidia shed during the first 5 days of the infection and by the significant decrease in the success of metamorphosis. The total number of glochidia that successfully attached to the fish decreased significantly during the fourth infection relative to the first. The number of larvae attached to the host fish was positively correlated with the size of the fish during the first infection but was negatively correlated during all subsequent infections. Variance to mean ratios indicated that larvae were aggregated among host fishes during the infections. This study has important implications in propagation and conservation efforts of this endangered group of organisms.

The identification of lectin-binding structures in adult worms of Echinococcus granulosus was carried out by lectin fluorescence; the distribution of carbohydrates in parasite glycoconjugates was also studied by lectin blotting. The lectins with the most ample recognition pattern were ConA, WGA, and PNA. ConA showed widespread reactivity in tegument and parenchyma components, including the reproductive system, suggesting that mannose is a highly expressed component of the adult glycans. Although reproductive structures appeared to be rich in N-acetyl-d-glucosamine (GlcNAc)–N-acetyl neuraminic acid (NeuAc) and galactose (Gal) as demonstrated by their strong reactivity with WGA and PNA, respectively, some differences were observed in their labeling patterns. This was very clear in the case of the vagina, which only reacted with WGA. Furthermore, WGA and ConA both had reactivity with the excretory canals. RCA, the other Gal binding lectin used, only reacted with the tegument, suggesting that widespread PNA reactivity with the reproductive system is related to the presence of the d-Gal-β-(1,3)-d-GalNAc terminal structure. UEA I failed to bind to any parasite tissues as determined by lectin fluorescence, whereas DBA and SBA showed a very faint staining of the tegument. However, in transferred glycans, N-acetyl-d-galactosamine (GalNAc) and fucose (Fuc) containing glycoproteins were distinctly detected.

Mosquitoes are vectors of many deadly and debilitating pathogens. In the current study, we used light and electron microscopies to study the immune response of Aedes aegypti hemocytes to bacterial inoculations, Plasmodium gallinaceum natural infections, and latex bead injections. After challenge, mosquitoes mounted strong phagocytic and melanization responses. Granulocytes phagocytosed bacteria singly or pooled them inside large membrane-delimited vesicles. Phagocytosis of bacteria, Plasmodium sporozoites, and latex beads was extensive; we estimated that individual granulocytes have the capacity to phagocytose hundreds of bacteria and thousands of latex particles. Oenocytoids were also seen to internalize bacteria and latex particles, although infrequently and with low capacity. Besides phagocytosis, mosquitoes cleared bacteria and sporozoites by melanization. Interestingly, the immune response toward 2 species of bacteria was different; most Escherichia coli were phagocytosed, but most Micrococcus luteus were melanized. Similar to E. coli, most Plasmodium sporozoites were phagocytosed. The immune response was rapid; phagocytosis and melanization of bacteria began as early as 5 min after inoculation. The magnitude and speed of the cellular response suggest that hemocytes, acting in concert with the humoral immune response, are the main force driving the battle against foreign invaders.

Prior exposure of Biomphalaria glabrata to the eggs of an incompatible digenean, Plagiorchis elegans, rendered this snail host less suitable to a compatible species, Schistosoma mansoni. Although P. elegans failed to develop patent infections in B. glabrata, it reduced the production of S. mansoni cercariae by 88%. Concomitantly, host attributes such as reproduction, growth, and survival were compromised. The effect of P. elegans infection was most severe among snails that, in addition, had developed patent schistosome infections. Although few S. mansoni cercariae were produced, egg production by B. glabrata was only 4% of control values. Furthermore, no doubly infected snails survived for more than 3 wk after patency, whereas controls experienced no mortality during the same time period. The above effects were attributable to the establishment and persistence of P. elegans sporocysts in the tissues of the incompatible snail host. Their indirect antagonistic interaction with the larval stages of S. mansoni may be mediated, in part, through their long-term stimulation of the host's internal defense mechanisms. These findings are discussed with a view to use P. elegans and other plagiorchiid digeneans as agents in the biological control of snails and snail-borne diseases.

The objective of this work was to examine the calcium concentration in the shells of naturally infected individuals of Lymnaea stagnalis. Calcium concentration in the water of lakes and in snail shells was examined by the EDTA method. Calcium concentration in the shells of infected snails was found to be higher than that in the shells of uninfected ones only in the lake with the lowest calcium concentration in water. However, this difference is not evidence of hypercalcification of snail shells caused by trematode larvae but suggests that the limited calcium concentration in the environment can be a very good factor for studying snail shell calcification under natural conditions. Moreover, small adult snails from 2 lakes had more calcium in their shells than did large ones. Different reproductive rates of different-sized snails could be the cause of this phenomenon.

This study addressed the problem of local patterns of host specificity among Ancyrocephalinae (Monogenoidea) on bass and sunfish species, when the hosts occur in different species combinations in separate ponds. One hundred fifty-three fish of the Centrarchidae, from 4 study sites in Nebraska, were collected. Host species included bluegill (Lepomis macrochirus), green sunfish (L. cyanellus), largemouth bass (Micropterus salmoides), black crappie (Pomoxis nigromaculatus), white crappie (P. annularis), and rock bass (Ambloplites rupestris). These fish occurred in different species combinations, depending on the pond sampled. Results indicated that several centrarchid species could inhabit the same pond and yet support distinct monogene communities. Clavunculus bursatus, Onchocleidus helicis, O. principalis, and Syncleithrum fusiformis were found only on largemouth bass, regardless of what other centrarchids were present in a particular pond. Haplocleidus dispar occurred on green sunfish, bluegill, largemouth bass, and black crappie, and H. furcatus occurred on both bluegill and largemouth bass. Onchocleidus cyanellus and O. ferox were found on both bluegill and green sunfish. Rock bass were present in only 1 of the 4 ponds, but were not infected with any monogenes, even though co-occurring centrarchids were often heavily infected. Largemouth bass had the most diverse ancyrocephaline communities. The degree of parasite host specificity among these monogenes was inversely related to the diversity of host species present in a particular pond. In general, the parasites were more host specific than might be inferred from the literature; parasite species did not necessarily colonize supposedly receptive host species even when the latter were present, and host relatedness was the major factor in determining whether host species shared a common parasite species.

To determine the potential for dissemination of myxozoan parasites by transfer of their alternate oligochaete hosts, shipments of tubificid worms obtained from an overseas commercial aquarium supplier were screened for actinospore stages of myxozoan parasites. At least 7 different triactinomyxon types were identified. The morphological characteristics of the actinospores recovered from these tubificids shared characteristics with triactinomyxons characterized in other surveys, particularly from eastern Europe. Analysis of the screened samples by polymerase chain reaction and comparison of morphological data indicated that these actinospores did not correspond to the triactinospore of Myxobolus cerebralis. Although identification of these triactinomyxon types was beyond the scope of this study, these data suggest that the unregulated import–export and exchange of live organisms for ornamental fish food may result in accidental introduction or dissemination of myxozoan parasites.

A serine proteinase gene was isolated from a cDNA library of Trichinella spiralis muscle larvae at 30 days postinfection (PI). The library was immunoscreened with T. spiralis–infected sera. A clone, designated Ts23-2, contained a cDNA transcript of 1,445 bp that encoded a putative signal peptide of 27 amino acids, a proregion of 20 amino acids, and a predicted mature enzyme of 374 amino acids. The predicted molecular mass of the Ts23-2 mature protein was 42.3 kDa. The enzyme comprised 2 regions, a catalytic domain of 234 residues and a C-terminal domain. The closest homologues of the Ts23-2 mature protein were serine proteinases from a wide range of organisms. The catalytic domain of the Ts23-2 clone was expressed as a proform in Escherichia coli. The recombinant protein cleaved serine proteinase–specific synthetic peptide substrates, and class-specific inhibitors of serine proteinases inhibited the enzymatic activity. Antibody against the Ts23-2 recombinant protein stained proteins migrating at about 51 and 33 kDa in crude extracts from 30-day PI muscle larvae and 18-day PI muscle larvae, but it failed to stain any proteins in crude extracts from newborn larvae and adult worms or in excretory–secretory products from 30-day PI muscle larvae. Production of the mRNA transcript for the Ts23-2 gene was mainly restricted to the 30-day PI muscle larvae, suggesting stage-specific expression. Intense staining with the anti–Ts23-2 serum was found within the parasites at the muscle stage of 30 days PI.

Recently, several investigators have reported large-bowel diarrhea in cats associated with intestinal trichomonad parasites. These reports have presumptively identified the flagellates as Pentatrichomonas hominis, an organism putatively capable of infecting the intestinal tracts of a number of mammalian hosts, including cats, dogs, and man. The purpose of the present study was to determine the identity of this recently recognized flagellate by means of rRNA gene sequence analysis; restriction enzyme digest mapping; and light, transmission, and scanning electron microscopy (SEM).

Trichomonad total extracts (TTE), or vesicular (P30) and soluble (S30) subcellular fractions from 3 pathogenic Trichomonas vaginalis strains (GT-3, GT-13, and GT-15), lysed both human and Sprague–Dawley rat erythrocytes in a time- and dose-dependent manner. The entire hemolytic activity of TTE was located in P30, showing 2 peaks of maximum activity, one at pH 6.0 and another at pH 8.0, in the presence of 1 mM Ca² . Hemolytic activity on rat erythrocytes was greater at pH 6.0 (6.71 ± 0.33 hemolytic units [HU]/mg/hr to 11.60 ± 0.24 HU/mg/hr) than at pH 8.0 (3.81 ± 0.30 HU/mg/hr to 5.75 ± 0.65 HU/mg/hr), and it was greater than that on human red blood cells at pH 6.0 (2.67 ± 0.19 HU/mg/hr to 4.08 ± 0.15 HU/mg/hr) or pH 8.0 (2.24 ± 0.09 HU/mg/hr to 2.81 ± 0.06 HU/mg/hr). The alkaline and acidic hemolytic activity diminished (60–93% at pH 6.0 and 78–93% at pH 8.0) by the effect of 80 μM Rosenthal's inhibitor, which also inhibited 27–45% and 29–54% trichomonad alkaline and acidic phospholipase A activities, respectively. Vesicles, vacuoles, and hydrogenosomes were rich in P30. Trichomonas vaginalis has a hemolytic PLA, which could be involved in its cytopathogenic mechanism.

Rhabdias anolis n. sp. from the lungs of Anolis frenatus collected in Panama is described and illustrated. Rhabdias anolis n. sp. represents the 45th species assigned to the genus and the ninth from the Neotropical Realm. It is distinguished from all other Neotropical species by the presence of a pair of sessile lateral papillae on its tail.

Strongyluris panamaensis n. sp. from the intestine of Anolis biporcatus collected in Panama is described and illustrated. Strongyluris panamaensis represents the 30th species assigned to this genus and the fourth from the neotropical region. Strongyluris panamaensis can be easily distinguished from all other neotropical species because it has the shortest spicule. In addition to S. panamaensis, 3 species of Trematoda, Mesocoelium monas, Parallellopharynx arctus, and Urotrema scabridum, and 3 species of Nematoda, Crytosomum penneri, Oswaldocruzia bainae, and Piratuba digiticauda, were identified.

A new genus of Nippostrongylinae, Malvinema n. gen., with 3 coparasitic species M. frederici n. sp., M. carolinae n. sp., and M. victoriae n. sp. from the intestine of the water rat, Scapteromys aquaticus Thomas (Rodentia: Muridae), from the northeast of Buenos Aires Province, Argentina, is proposed in this study. The new genus shows similarities to 2 Neotropical Nippostrongylinae: Carolinensis (Travassos, 1937) by some characters of the synlophe and Stilestrongylus Freitas, Lent and Almeida, 1937, by the pattern of the caudal bursa. It is characterized by a synlophe with triple or quadruple gradient of size of the ridges, lateromedian, decreasing from the largest left and right ridges. The gradient situated in the right ventral quadrant is always present. The caudal bursa shows a pattern of type 1-4. Malvinema frederici possesses a synlophe with 17 ridges and an axis of orientation inclined at 45° from the sagittal axis; M. carolinae possesses a synlophe with 22–24 ridges and an axis of orientation almost merged with the sagittal axis. Both species have a caudal bursa with the right lobe enlarged transversally. Malvinema victoriae possesses a synlophe with 22–24 ridges, an axis of orientation inclined at 45° from the sagittal axis, and a caudal bursa with the right lobe enlarged vertically.

The original description of the type species of Allencotyla, A. mcintoshi Price, 1962, was based on 5 specimens collected from 1 amberjack Seriola lalandi in Florida. In this study, A. mcintoshi is redescribed based on new specimens collected from juvenile greater amberjacks S. dumerili captured off the Spanish Mediterranean coast and maintained in experimental culture tanks. Except for a slightly smaller size, individuals of S. dumerili were morphologically similar to the type specimens. However, reexamination of the type material and examination of the new specimens revealed the following discrepancies with respect to the original description: (1) the vaginal pore is unarmed and covered with pointed folds with a spinelike appearance; (2) the cirrus is armed, but spines are frequently lost (e.g., in the type specimens); (3) the esophagus is long with 3 or 4 pairs of diverticula and branches into cecae at the level of the genital atrium; and (4) the genital atrium is surrounded by 8 concentric crowns of spines, not 11 or 12 as reported in the original description. An emended diagnosis of Allencotyla is presented.

A new species of parasitic nematode, Cystoopsis atractostei (Trichinelloidea: Cystoopsidae), is described based on female specimens recovered from the subcutaneous tissue of the tropical gar, Atractosteus tropicus Gill (Lepisosteiformes: Lepisosteidae), from 2 localities (Canal Nueva Esperanza and Canal Tabasquillo) of the Pantanos de Centla Biological Reserve, State of Tabasco, southeastern Mexico, collected in April 2001. The total prevalence was 13%, and the mean intensity of infection was 1 nematode per fish. The new species differs from females of the only other adequately described congeneric species, C. acipenseris Wagner, 1867, mainly in possessing a bulbous inflation at the anterior end of the muscular esophagus, the vulva situated well posterior to the nerve ring, smooth cuticle, and in the shape of the posterior vesicular portion of the body (markedly transversely oval) in the largest specimens. Both species also differ in their host types (Lepisosteiformes vs. Acipenseriformes) and in geographical distribution (tropical southern Mexico vs. temperate zones of the Holarctic).

Auriculotrema lechneri n. gen., n. sp. is described from the small intestine of Emydura krefftii and Elseya latisternum from northern Queensland, Australia. The new species strongly resembles species of Choanocotyle in body shape, ventral incision of the oral sucker, structure of the cirrus sac, and location of the genital pore. The distinctive taxonomic feature is the presence of 2 winglike projections extending beyond the lateral margins of the oral sucker, in contrast to the extremely large, expanded oral sucker diagnostic of Choanocotyle spp. Auriculotrema n. gen. is the second genus included in the formerly monotypic Choanocotylidae Jue Sue and Platt, 1998.

Choanocotyle hobbsi n. sp. and Choanocotyle juesuei n. sp. are described from the small intestine of the oblong turtle Chelodina oblonga from the vicinity of Perth, Western Australia. These are the third and fourth species referred to Choanocotyle. Choanocotyle hobbsi is most similar to Choanocotyle nematoides but differs in the size and shape of the oral sucker and the absence of a median loop in the cirrus sac. Choanocotyle juesuei is most similar to Choanocotyle elegans but differs in the size of the oral sucker and other morphometric criteria. Comparative analysis of the sequences of different nuclear ribosomal deoxyribonucleic acid regions of C. nematoides and C. hobbsi has confirmed that they are closely related but distinct species.

Development stages of a haemogregarine were found in Giemsa-stained heart blood smears of 3 of the 4 horned blennies (Parablennius cornutus) captured at De Hoop Nature Reserve, South Africa. Gamonts of this haemogregarine conformed to an existing description of Desseria (Haemogregarina) fragilis from P. cornutus, but intraerythrocytic trophozoites, as well as meronts undergoing division, were reported for the first time for this species. A detailed redescription of D. fragilis allowed its taxonomic re-evaluation. Intraerythrocytic division excluded D. fragilis from Desseria, whereas morphometric similarities and identical patterns of development lead to the conclusion that D. fragilis is the cosmopolitan haemogregarine, Haemogregarina bigemina. Nomenclatural correction synonymizing the 2 species is recommended.

Acanthotrema felis n. sp. (Digenea: Heterophyidae) is described from adult flukes recovered in the small intestine of stray cats caught in the southwestern part of the Republic of Korea. The worms were characterized by minute body size, the presence of a bipartite seminal vesicle, and a ventral sucker associated with a ventrogenital sac, which enclosed 3 sclerites (2 long and pointed, and 1 short and thumblike), and an unarmed gonotyl. They were assigned to Acanthotrema Travassos, 1928, which currently contains 4 species. The new species resembles A. acanthotrema, the type species, because both have 3 sclerites on the ventrogenital sac. However, all 3 sclerites in the new species are armed with minute spines on their base, whereas only 2 sclerites in A. acanthotrema have minute spines near their tips. Three species of Stictodora Looss, 1899, namely, Stictodora tridactyla Martin and Kuntz, 1955, S. cursitans Kinsella and Heard, 1974, and S. tanayensis Velasquez, 1973, are transferred to Acanthotrema, and the new species is distinguished from them particularly in terms of the number and shape of its sclerites. The generic diagnosis of Acanthotrema is emended.

Acanthostomum macroclemidis n. sp. is described from specimens found in the intestine of an alligator snapping turtle Macroclemys temmincki from southern Mississippi. The most important diagnostic features of the new species are the general shape and proportions of the body, the position of the pharynx (relative length of the prepharynx and esophagus), the egg size, the relative length and position of the vitelline fields, and the number, shape, and size of the circumoral spines. The new species has a very elongated body (length–width ratio, 8.9–13.0:1), 26 circumoral spines, which are almost oval in shape, a long prepharynx and a very short (shorter than the pharynx) esophagus, a seminal receptacle situated between the ovary and the anterior testis, a uterus not extending posterior to the anterior margin of the ovary, a long-stemmed and short-armed excretory vesicle, and 2 anal openings. Some features of the external morphology, such as the suckers, circumoral spines, sensory papillae, tegumental spines, and morphology of the posterior end, are examined using scanning electron microscopy. A diagnosis differentiating A. macroclemidis n. sp. from some other acanthostomine digeneans is provided. Acanthostomum macroclemidis n. sp. is the first digenean reported from an alligator snapping turtle and represents the northernmost record of an acanthostomine from turtles.

Two unusual myxozoan parasites are described from the somatic muscle of 2 reef fishes from Australia's Great Barrier Reef. Kudoa quadricornis n. sp. from the somatic muscle of Carangoides fulvoguttatus is morphologically consistent with other Kudoa sp., having 4 polar capsules and 4 shell valves. Kudoa quadricornis n. sp. is unique in that it has a pyriform spore body with a greater length than width (7.82–9.95 and 5.94–8.66 μm, respectively) and distinct posterolateral projections. Spores of Kudoa permulticapsula n. sp. observed within pseudocysts of the somatic muscle tissue of Scomberomorus commerson are different from those of all other myxozoans. The ovoid spores (length, 4.69–6.65 μm; width, 8.42–9.92 μm; thickness, 6.36–8.33 μm) contain 13 polar capsules with an equal number of shell valves. Phylogenetic analysis using small subunit ribosomal DNA sequences of K. quadricornis n. sp. and K. permulticapsula n. sp. showed that these parasites cluster within a clade comprised of Kudoa species. This brings into question the division of parasites of the Multivalvulida into genera based solely on polar capsule numbers.

Baylisascaris procyonis, raccoon roundworm, causes a severe retinal lesion in humans. The lesion is termed as diffuse unilateral subacute neuroretinitis (DUSN). To understand the pathogenesis of B. procyonis in gerbils, we inoculated 17 embryonated eggs/g body weight of B. procyonis into 15 male Mongolian gerbils, Merionis ungiculatus, and monitored their fundi with an ophthalmoscope. Six of 15 gerbils (40%) showed severe retinitis with a sinuous track due to larval movement. The lesions extended across nearly half of the affected fundi. Histopathological examination revealed perivasculitis in the optic disk region, inflammatory proliferation of the pigment cells, and vitreitis in most cases. These findings were similar to those in human cases of DUSN, suggesting that gerbils might be a useful model for understanding the pathogenesis of B. procyonis infection in humans.

Chicks were experimentally infected with Acanthoparyphium tyosenense (Digenea: Echinostomatidae) metacercariae per os, and the growth and development of worms in this host were observed from days 1 to 38 postinfection (PI). The worms grew rapidly and matured sexually in the small intestine (chiefly in the jejunum) of chicks by day 5 PI, and survived at least up to day 38 PI, although worm recovery decreased after day 5 PI. Both parenchymal and reproductive organs increased greatly in size from day 2 to day 10 PI and then continued to increase gradually in size up to day 38 PI. The number of uterine eggs reached a peak on days 10 and 15 PI and then decreased gradually. The results suggest that chicks are a fairly suitable definitive host for experimental infection with A. tyosenense.

Monoclonal antibodies (mAbs) were produced against the proteins of advanced third-stage larvae (AdL3) of Gnathostoma nipponicum. Six mAbs (Gn2C3, Gn2H3, Gn4C3, Gn4E9, Gn5H1, and Gn10B7) were obtained as determined by enzyme-linked immunosorbent assay (ELISA). Gn4E9 and Gn5H1 seemed to be genus-specific, as they did not cross-react with Anisakis sp., Dirofilaria immitis, Gongylonema pulchrum, Toxocara canis, Trichinella sp., Trichuris vulpis, Metagonimus sp., or Spirometra erinaceieuropaei by ELISA. Immunohistochemistry showed that Gn2C3, Gn4E9, and Gn5H1 reacted strongly with the central esophagus; Gn2H3 reacted with cuticle, muscle, intestine, and the cervical sac; and Gn4C3 and Gn10B7 reacted with cuticle, muscle, esophagus, intestine, and the cervical sac of AdL3. In Western blotting analysis, Gn2C3, Gn4E9, and Gn5H1 reacted to 60-, 53-, 46-, and 41-kDa proteins; Gn4C3 reacted to the AdL3 protein of G. nipponicum (>42 kDa). Moreover, proteins purified using a mAb Gn4E9 immunoprecipitation method (sizes 60-, 53-, 46-, and 41-kDa) were used as antigens in ELISAs. A significant difference (P < 0.01) was shown between mouse sera infected with G. nipponicum and sera infected with Trichnella sp. or not infected. These results provide a rationale for evaluating esophageal proteins for the development of diagnostic methods for detecting G. nipponicum or Gnathostoma sp. infections.

Species of Philophthalmus parasitize primarily the eyes of wild and domestic birds. A variety of mammals, including humans, occasionally serve as the intermediate as well as the definitive hosts for this parasite, although human cases are extremely rare. Here, we report a case of human conjunctivitis caused by an infection with Philophthalmus sp. in Mexico. The patient was a 31-yr-old male who visited an ophthalmologist in Los Mochis, Sinaloa, Mexico, because of a foreign-body sensation in his left eye for 2 mo. A small live parasite was found in the connective tissue of the bulbar conjunctiva and was removed surgically under local anesthesia under ophthalmoscopic observation. The parasite was identified morphologically as Philophthalmus lacrimosus Braun, 1902. This is the first case of human philophthalmosis in Mexico and, to our knowledge, the first human case of P. lacrimosus infection in the world.

To determine how long waterborne spores of Encephalitozoon cuniculi, E. hellem, and E. intestinalis could survive at environmental temperatures, culture-derived spores were stored in water at 10, 15, 20, 25, and 30 C and tested for infectivity in monolayer cultures of Madin Darby bovine kidney (MDBK) cells. At 10 C, spores of E. intestinalis were still infective after 12 mo, whereas those of E. hellem and E. cuniculi were infective for 9 and 3 mo, respectively. At 15 C, spores of the same species remained infective for 10, 6, and 2 mo, and at 20 C, for 7, 5, and 1 mo, respectively. At 25 C, spores of E. intestinalis and E. hellem were infective for 3 mo, but those of E. cuniculi were infective for only 3 wk. At 30 C, the former 2 species were infective for 3 wk and 1 mo, respectively, and the latter species for only 1 wk. These findings indicate that spores of different species of Encephalitozoon differ in their longevity and temperature tolerance, but at temperatures from 10 to 30 C, all 3 have the potential to remain infective in the environment long enough to become widely dispersed.

Infection with helminth parasites typically evokes a T helper 2-cell response in the mammalian host. Interleukin (IL)-4 and IL-13 are important in the rapid expulsion of parasitic nematodes, with the absence of either cytokine being compensated for by the other. However, the IL-4 and IL-13 common signaling molecule, signal transducer and activator of transcription 6 (STAT-6) appears to be mandatory for the spontaneous expulsion of enteric helminths. Mice genetically deficient in IL-4, IL-13, or STAT-6 were infected with the cestode Hymenolepis diminuta and worm infectivity and jejunal goblet cell responses assessed 12–18 days postinfection (PI). Only the STAT-6 knockout (KO) animals harbored adult worms; neither gravid adult nor stunted H. diminuta was obtained from the infected IL-4 KO or IL-13 KO mice ≥12 days PI. Also, the establishment of worms in the intestine of STAT-6 KO animals was associated with a reduced goblet cell response. These findings support the hypothesis that increased mucin production is an important part of the host response to tapeworm infection and that functional STAT-6 signaling may be an absolute requirement for the rejection of intestinal cestodes and thus, helminth parasites in general.

Gametocyte sex ratio of the malaria parasite Plasmodium mexicanum is variable in its host, the western fence lizard (Sceloporus occidentalis), both among infections and within infections over time. We sought to determine the effect of host physiological quality on the gametocyte sex ratio in experimentally induced infections of P. mexicanum. Adult male lizards were assigned to 4 treatment groups: castrated, castrated testosterone implant, sham implant, and unmanipulated control. No significant difference in gametocyte sex ratio was found among the 4 treatment groups. Two other analyses were performed. A surgery stress analysis compared infection sex ratio of castrated, castrated testosterone implant, and sham implant groups with the unmanipulated control group. A testosterone alteration analysis compared infection sex ratio of the castrated and castrated testosterone implant groups with the sham implant and unmanipulated control groups. Again, no significant difference was observed for these 2 comparisons. Thus, physiological changes expected for experimentally induced variation in host testosterone and the stress of surgery were not associated with any change in the gametocyte sex ratio. Also, the experiment suggests testosterone is not a cue for shaping the sex ratio of gametocytes in P. mexicanum. These results are related to the evolutionary theory of sex ratios as applied to malaria parasites.

Biomphalaria peregrina was found to be naturally infected with cercariae of Zygocotyle lunata in a pond of the Zoological Garden of Buenos Aires. Mice and chicks were fed metacercariae and gravid adults recovered. Eggs recovered from mice feces were used for experimental infections. Laboratory-reared B. peregrina and 4 other Biomphalaria spp. were successfully infected with Z. lunata miracidia. Biomphalaria glabrata was refractory. Species of Helisoma, the only intermediate hosts of Z. lunata so far reported, have never been found in Argentina. Species of Biomphalaria may be intermediate hosts of Z. lunata in the southern region of the Paraná River Basin.

To assess the potential risk for tick-borne agents, Ixodes ricinus were collected from 2 sites in northwestern Poland. The ticks were tested by polymerase chain reaction for coinfection with Borrelia burgdorferi sensu lato (s. l.), human granulocytic ehrlichiosis (HGE) agent, and Babesia microti. Of the 533 processed ticks, 16.7% were positive for B. burgdorferi s. l., 13.3% for B. microti, and 4.5% for the HGE agent. Twenty ticks were coinfected with 2 or 3 of the pathogens.

Partial mitochondrial 16S ribosomal ribonucleic acid gene sequences in the ticks Carios capensis collected from black-footed albatross, Diomedea nigripes, colonies on Torishima Island, Japan (30°28′N, 140°18′E), were examined. The sequence was compared with those of C. capensis from Hawaii, South Carolina, and Texas. The sequences were all identical in ticks from Torishima and 2 from Hawaii. There were 2–3 transitions between the other Hawaiian and Texas ticks and Torishima specimens. Two transitions were also observed when compared with the ticks from South Carolina. The results suggest the possibility of gene flow between tick populations at each of the 2 geographic sites, which probably was accomplished by tick-infested migratory seabirds at their breeding sites. Sequence comparison analysis indicated that the C. capensis ticks are on the branch with C. marginatus and C. mexicanus ticks and not with Ornithodoros. This supports the revision suggested by Klompen and Oliver (1993).

Diplozoidae monogeneans are fish-gill ectoparasites comprising 2 individuals fused in so-called permanent copula. This unique situation occurs when 2 larvae (diporpae) make contact on the host gill, such that their union triggers maturation into an individual adult worm. The present study examined paired stages of Eudiplozoon nipponicum microscopically to ascertain whether somatic fusion involves neural connectivity between these 2 heterogenic larvae. Neuronal pathways were demonstrated in whole-mount preparations of the worm, using indirect immunocytochemical techniques interfaced with confocal scanning laser microscopy for peptidergic and serotoninergic innervations and enzyme cytochemical methodology and light microscopy for cholinergic components. Elements of the central nervous systems of paired worms are connected by commissures in the region of fusion so that the 2 systems are in structural continuity. Interindividual connections were most apparent between corresponding ventral nerve cords. All 3 classes of neuronal mediators were identified throughout both central and peripheral connections of the 2 nervous systems. The anatomical complexity and apparent plasticity of the diplozoon nervous system suggest that it has a pivotal role not only in motility, feeding, and reproductive behaviors but also in the events of larval pairing and somatic fusion.