Showing papers in "Journal of Parasitology in 1990"

Effect of high temperature on infectivity of Toxoplasma gondii tissue cysts in pork.

Abstract: To study the effect of high temperature on infectivity of Toxoplasma gondii tissue cysts, pork from infected pigs was mixed with infected mouse brains and homogenized thoroughly. Twenty-gram samples of infected homogenized meat were sealed in plastic pouches, pressed to a uniform thickness of 2 mm, and subjected to water-bath temperatures of 49, 52, 55, 58, 61, 64, and 67 C for 0.01, 3, 6, 9, 12, 24, 48, and 96 min. Treated samples were digested in HCl-pepsin solution and bioassayed in mice. Toxoplasma gondii tissue cysts remained viable at 52 C for 9.5 min but not for 9.5 min at 58 C; tissue cysts were generally rendered nonviable by heating to 61 C or higher temperature for 3.6 min. Tissue cysts survived once at 64 C for 3 min. These data demonstrate that T. gondii tissue cysts are less heat resistant than encysted Trichinella spiralis larvae.

Fatal congenital Neospora caninum infection in a lamb.

Abstract: Neospora caninum tissue cysts were found in the brain and spinal cord of a 1-wk-old lamb that was unable to stand after birth. The lamb was originally diagnosed as having toxoplasmosis, but ultrastructural and immunohistochemical techniques used in the present study permitted a definitive diagnosis of Neospora caninum tissue cysts in the brain and spinal cord of this lamb. This is the first report of N. caninum in sheep.

Efficacy of agar-plate culture in detection of Strongyloides stercoralis infection.

Abstract: Agar-plate culture of feces using a modified petri dish proved to be highly efficient in the detection of Strongyloides stercoralis infection Furrows left by S stercoralis on the agar plate were distinguished readily in size from those left by Necator americanus

Neospora caninum (Apicomplexa) in an aborted equine fetus

Abstract: Tachyzoites of Neospora caninum were found in sections of lung of an equine fetus aborted 2 mo before term. Individual tachyzoites were approximately 3-5 x 2-3 microns, divided by endodyogeny, and stained positively with anti-N. caninum serum but not with anti-Toxoplasma gondii serum. Toxoplasma gondii antibody was not found in the mare's serum. This is the first report of N. caninum in a horse and indicates that N. caninum can be transmitted transplacentally in equids.

Axenic culture and characterization of Giardia ardeae from the great blue heron (Ardea herodias).

Abstract: Trophozoites of Giardia ardeae were obtained from the great blue heron (Ardea herodias) and established in axenic culture using the TYI-S-33 medium. The generation time in culture for G. ardeae was 22-25 hr, which was 3-fold longer than for Giardia duodenalis (WB strain). A morphological comparison of trophozoites in the original intestinal isolate to those grown in culture revealed that they were identical for the following characteristics: a pyriform-shaped body, a ventral adhesive disc with a deep notch in the posterior border, teardrop-shaped nuclei, pleomorphism in median body structure ranging from a round-oval appearance (Giardia muris type) to that of a clawhammer (G. duodenalis type), and a single caudal flagellum on the right side (as viewed dorsally) with the left one being rudimentary. Analysis of the chromosomal migration patterns was performed by orthogonal-field-alternation gel electrophoresis and demonstrated that the pattern for G. ardeae was distinctly different from that for G. duodenalis (Portland 1-CCW strain). Bacterial symbionts were seen attached to trophozoites in the original isolate but could not be detected in cultured trophozoites using scanning electron microscopy, fluorescence light microscopy using the Hoechst 33258 dye for DNA localization, or by standard microbiological techniques using nonselective media for growing aerobic or anaerobic bacteria. This study demonstrated that avian-derived Giardia could be grown in axenic culture; based on morphological criteria and chromosomal migration patterns, that G. ardeae should be considered a distinct species; and that rationale for determining Giardia spp., based on median body structure alone, should no longer be considered adequate for classification at the species level.

In vitro attenuation of Cryptobia salmositica and its use as a live vaccine against cryptobiosis in Oncorhynchus mykiss.

Abstract: r: The present communication reports on the attenuation of a pathogenic hemoflagellate, Cryptobia salmositica Katz (Sarcomastigophora: Kinetoplastida) and its use as a live vaccine against cryptobiosis. The parasite was attenuated by continuous in vitro culture (at 10 C for 55 wk) in minimum essential medium. Attenuated (culture) forms are morphologically similar to virulent (blood) forms. They are however more slen- der and have a shorter anterior flagellum and a smaller nucleus and kinetoplast. The attenuated form returned to its normal size and multiplied when inoculated into naive Oncorhynchus mykiss. It produced a low para- sitemia but did not cause disease (e.g., no exophthalmia or anemia) in fish. At four wk after infection, the vac- cinated fish were challenged with the virulent parasite. They were protected from the disease, whereas the con- trol (naive) fish, infected with only the virulent parasite, had the usual clinical signs (e.g., anemia, exophthal- mia). No parasite was detected in any of 10 vaccinated fish at 22 wk after challenge with the virulent parasite. However, 5 of 9 fish infected with culture forms and 6 of 9 fish infected with blood forms still had detectable parasites at 26 and 22 wk after infection, respectively. Cryptobia salmositica is 1 of the 2 pathogenic hemoflagellates known to cause disease in eco- nomically important fishes in North America; it has been recorded from all species of Pacific salmon (see Woo, 1987a). The clinical signs of the disease include exophthalmia, splenomegaly, edema, abdominal distension with ascites fluid, a microcytic and hypochromic anemia (Woo, 1979), immunodepression (Wehnert and Woo,

Effects of murine giardiasis on growth, intestinal morphology , and disaccharidase activity.

Abstract: The aim of this study was to assess the effects of Giardia muris on host growth and food intake, small intestinal morphometrics, mucosal enzyme activities, and brush border ultrastructure. Weanling mice infected with 1,000 G. muris cysts were compared to control and pair-fed sham-treated animals. Infection with G. muris resulted in decreased food intake and retarded growth. In infected animals, villus atrophy was observed in the duodenum throughout the study period and in the jejunum on days 8 and 50. On day 30, whereas jejunal architecture returned to normal in infected animals, malnourished pair-fed animals exhibited a compensatory increase in villus height. Sucrase and maltase were depressed in infected animals on days 2-24. On day 8 jejunal disaccharidases in pair-fed animals were also decreased but to a lesser extent than in infected animals. On day 24, disaccharidase values for control and infected mice were similar, whereas values in pair-fed animals were increased. On day 8, jejunal microvilli were shorter in infected animals than in control and pair-fed animals. This brush border injury was present throughout the jejunum and was also observed in pair-fed animals, but to a lesser extent. These findings suggest that G. muris retards growth in weanling mice, results in small intestinal injury, and interferes with the compensatory response to malnutrition of the infected host. Villus atrophy and brush border enzyme deficiencies associated with the disease mainly occur in the duodenum and jejunum, where trophozoites are most numerous. In infected and in pair-fed animals, the decrease in jejunal disaccharidase activities correlated with a diffuse shortening of brush border microvilli.

Serum-stimulated feeding in vitro by third-stage infective larvae of the canine hookworm Ancylostoma caninum.

Abstract: ABsTRAcr: Developmentally arrested nonfeeding infective larvae of hookworms resume development after entry into the host, presumably in response to a signal encountered during invasion. Logically, an initial step in the resumption of development might be the resumption of feeding. An in vitro assay for feeding is described for the third-stage larvae of the canine hookworm Ancylostoma caninum. Populations of larvae incubated under hostlike conditions in the presence of 10% canine serum resume feeding within 6 hr, as evidenced by the uptake of fluorescein-labeled bovine serum albumin. Feeding is dependent on the presence of canine serum, and peaks by 24 hr incubation. Maximal feeding levels occur at temperatures above 34 C with a gas phase of 5% CO2/ 95% air, whereas culture medium and pH are unimportant for feeding. Serum concentrations between 0.1% and 1.0% (v/v) initiate feeding, and the response peaks at approximately 8.0% serum. Serum triggers feeding within 6 hr and is not required for feeding to continue once initiated. The saturation effect and the trigger phenomenon suggest that the initiation of feeding is a receptor-mediated response.

Prevalence and abundance of equine strongyles (Nematoda: Strongyloidea) in tropical Australia.

Abstract: A postmortem survey of 57 horses in tropical northern Queensland revealed 41 (89%) infected with intestinal strongyles. Thirty-five strongyle species (8 large strongyles and 27 small strongyles [Cyathostominae]) were recorded of which 9 species are reported from Australia for the first time. The 14 most prevalent small strongyles were Cyathostomum catinatum (in 76% of horses), Cyathostomum coronatum (65%), Cyathostomum pateratum (33%), Cyathostomum labiatum (30%), Cylicostephanus calicatus (70%), Cylicostephanus longibursatus (67%), Cylicostephanus goldi (43%), Cylicostephanus minutus (26%), Cylicocylus nassatus (67%), Cylicocyclus leptostomus (41%), Cylicocylus insigne (41%), Cylicocyclus radiatus (33%), Cylicocyclus brevicapsulates (22%), and Poteriostomum imperidentum (24%). The remaining cyathostomes were each found in less than 15% of horses. The 4 most common large strongyles were Triodontophorus serratus (30%), Strongylus vulgaris (28%), Strongylus equinus, and Strongylus edentatus (both 22%). The number of species of small strongyles per horse showed a marked variation (mean 10.3, range 2-21) but bore no relationship to either the total number of strongyles per horse, age, sex, and breed of horse, or season. Total number of strongyles per horse (mean 15,890, range 20-165,000) was less than in recent surveys in Europe and the U.S.A. Most horses had low worm burdens, whereas a very small number were heavily infected. Ninety-seven per cent of the total strongyle counts were small strongyles. Strongylus species contributed just over 1%. Small numbers of large strongyles per horse were usual with T. serratus (mean 570), S. vulgaris (mean 330), and S. equinus (mean 330) the most numerous.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro effect of larval Schistosoma mansoni excretory-secretory products on phagocytosis-stimulated superoxide production in hemocytes from Biomphalaria glabrata.

Abstract: The in vitro production of the reactive oxygen metabolite superoxide (02-) was confirmed in hemocytes from the schistosome intermediate host Biomphalaria glabrata. Active forms of the enzyme super- oxide dismutase (SOD) inhibited reduction of nitroblue tetrazolium (NBT) to formazan in cells that had phago- cytozed zymosan particles, whereas an inactivated form of SOD did not. Moreover, based on the prevalence of 02 -positive hemocytes and the relative intensity of NBT staining reactions, hemocytes from the Schistosoma mansoni-resistant 10-R2 strain of B. glabrata possessed an overall greater capacity for generating superoxide than did those from S. mansoni-susceptible M-line snails. Schistosoma mansoni excretory-secretory (E-S) products, released during in vitro transformation of miracidia to sporocysts, inhibited phagocytosis of zymosan particles and superoxide activity in hemocytes from both snail strains, but 1 0-R2 hemocytes maintained higher levels of phagocytosis and superoxide production than did M-line hemocytes. The dose-dependent decreases in phagocytosis observed in both snail strains in the presence of E-S products could not account fully for the concomitant decrease in superoxide levels detected, indicating that either a single E-S factor differentially affects phagocytosis and superoxide production, or that different E-S factors are involved in the specific interference of each of these hemocyte functions. Immune compatibility of larval Schistosoma mansoni with the snail intermediate host Biom-

Identification of the secreted neutral proteases from Anisakis simplex.

Abstract: Ingestion of larval nematodes (family: Anisakidae) can cause the human disease known as anisakiasis. After ingestion, Anisakis larvae can be invasive, penetrating host stomach or intestinal wall. Observation of larvae penetrating the tissue layers of human stomach in vitro by SEM showed tunnels and burrows were formed in the mucosa and submucosa. Based on these observations, we hypothesized that secreted proteases may be involved in the degradation of host tissue macromolecules to allow tunnel formation. Using a model of connective tissue extracellular matrix (ECM), we found that as few as 5 Anisakis simplex larvae could degrade approximately 25% of the ECM in a 16-mm culture well in 24 hr. Further characterization of the secreted proteases using synthetic peptide substrates and inhibitors revealed that there were 2 classes of proteases present: a metallo aminopeptidase and a trypsinlike serine protease. Extracts of Anisakis larvae contained a 25-kDa protease that was recognized by rabbit anti-rat trypsin antibody on western blots. This suggests that there is structural as well as functional similarity between the Anisakis trypsin and vertebrate trypsins.

Infections in mice with tachyzoites and bradyzoites of Neospora caninum (Protozoa: Apicomplexa).

Abstract: Tachyzoites of 2 isolates of Neospora caninum (NC-1 and NC-2) were inoculated subcutaneously (s.c.), intraperitoneally (i.p.), or orally into mice to compare the effects of route of inoculation on pathogenicity. Mice developed more severe disease, and disease occurred sooner when inoculated with the NC-1 isolate compared to the NC-2 isolate. Deaths occurred earlier in mice inoculated i.p. with either isolate. Mice inoculated orally or s.c. with tachyzoites responded similarly to infection. Tissue cysts of the NC-2 isolate produced infections in mice following oral or s.c. inoculation. Lesions seen in mice inoculated with tachyzoites or bradyzoites were primarily acute pneumonia, myositis, encephalitis, ganglioradiculoneuritis, and pancreatitis. In vitro studies demonstrated that tachyzoites of both isolates were killed by incubation in pepsin-HCl solution but not 1% trypsin solution. Bradyzoites of the NC-2 isolate were able to withstand treatment with pepsin-HCl solution.

Anthelmintic activity of paraherquamide in sheep.

Abstract: Paraherquamide, an oxindole alkaloid metabolite of Penicillium paraherquei, was tested against the common gastrointestinal nematode species of sheep at 0.25, 0.5, 1.0, and 2.0 mg/kg, per os. It was highly efficacious (greater than or equal to 98% reduction) as a single oral treatment dosages greater than or equal to 0.5 mg/kg against adult Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis and Cooperia curticei, and the L4 stage of Cooperia spp. Noteworthy is the fact the isolate of H. contortus used was ivermectin-resistant and the isolate of T. colubriformis used was ivermectin- and benzimidazole-resistant. This suggests a different mode of action for paraherquamide relative to ivermectin and the benzimidazoles. The adult stage of Oesophagostomum columbianum was the dosage-limiting parasite with 79% efficacy recorded at the highest treatment level (2.0 mg/kg). Extrapolation from the O. columbianum response curve suggests a dosage in excess of 4.0 mg/kg would be required to attain 95% efficacy.

Effect of egg dosage and host genotype on liver trapping in murine larval toxocariasis.

Abstract: In this study we examined the effect of various initial sensitizing doses of infective Toxocara canis eggs and the effect of murine host genotype on the level of trapping of larvae in the liver after larval challenge. In the initial experiments, C57BL/6J mice were infected with a sensitization dose of 5, 25, 75, 125, or 250 infective T. canis eggs on day 0 postinfection (PI). On day 28 PI all mice were challenged with 500 infective eggs. On days 7, 14, and 21 postchallenge (PC) larval numbers within individual livers were determined. Trapping of larvae was observed in mice receiving a sensitization dose of 25 or more eggs. At 7 and 14 days PC the level of trapping increased with sensitization egg dose up to a dose of 125 eggs. At 21 days PC the level of trapping reached a plateau at a sensitization dose of 75 eggs. The peak level of larval trapping was observed on day 7 and day 14 PC following sensitization doses of 125 and 250 eggs, respectively. In the subsequent experiments, mice of various strains and H-2 haplotypes were inoculated with an initial sensitization dose of 125 eggs and a challenge dose of 500 eggs on day 0 and day 28 PI, respectively. Larval trapping within the liver was determined on day 14 PC. C57BL/6J mice trapped significantly more larvae than DBA/2J mice (P less than 0.01); all other strains trapped larvae at a lower, but statistically similar, level to the C57BL6/J mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction in cell entry of eimeria tenella (coccidia) sporozoites by protease inhibitors, and partial characterization of proteolytic activity associated with intact sporozoites and merozoites

Abstract: The role of proteases in the invasion of host cells by Eimeria tenella (Wisconsin strain) was studied in vitro. Protease inhibitors were used to treat sporozoites before inoculation or were applied to cultured chicken kidney cells before infection. The inhibitors antipain, leupeptin, aprotinin, L-l-tosylamide-2-phenyl-ethyl chlo- romethyl ketone (TPCK), or N-a-p-tosyl-L-lysine chloromethyl ketone (TLCK) reduced parasite invasion to 16-66% of control after treatment of cultured cells or sporozoites with 5- or 50-,g/ml concentrations of inhibitors in the culture medium. Phenylmethylsulfonyl fluoride (PMSF) reduced invasion to 32-57.7% at concentrations of 1-4 mM. The optimum pH for hydrolysis of azocasein by intact sporozoites or merozoites was determined over a range of pH 5.0 to pH 9.0. Sporozoites were highly active over a broad range from pH 5.5 to pH 9.0, with an apparent optimum at pH 8.0. Merozoites had a much lower specific activity with pH optima at 7.0 and 8.5. The protease activity of sporozoites or merozoites could be inhibited completely by the addition of 50 ,ug/ ml of leupeptin, TPCK, or TLCK or of 4 mM PMSF. Antipain inhibited proteases of sporozoites but not of merozoites. Pepstatin had little effect on either sporozoites or merozoites. The results suggest that parasite proteases of Eimeria may be necessary for invasion of host cells. The apicomplexan sporozoa possess an intri- cate system at the anterior of the sporozoite or merozoite called the apical complex that is im- portant to entry into the host cell. Associated with the apical complex is a system of rhoptries, saccules that empty their contents during cell en- try and during the rupture of the schizont (Jensen

Structure-activity relationships of benzothiazole and benzimidazole anthelmintics: a molecular modeling approach to in vivo drug efficacy.

Abstract: An investigation of the biochemical effects of an anthelmintic, tioxidazole (TIOX, methyl 6-[n-propoxy]benzothiazole-2-carbamate), on Hymenolepis diminuta in experimentally infected rats is reported. The chemotherapeutic actions of TIOX on H. diminuta in vivo were accompanied by marked changes in worm weight and chemical composition. Tapeworms recovered from rats that had received a therapeutically effective dose of TIOX 24 hr earlier were significantly smaller and contained much less glycogen (as a percentage of the wet weight) than worms from untreated controls. In TIOX-treated worms, protein concentrations rose at a rate sufficient to offset the decline in glycogen concentration. Glycogen/protein ratios in TIOX-treated worms were considerably lower than the corresponding control-values. Differences in the absolute amounts of glycogen and protein between control and drug-treated worms were even more pronounced. Administration of a subcurative dose of TIOX to the rat produced in H. diminuta another change, the onset of which preceded the gross alterations in worm weight and chemical composition. In vitro studies, carried out 18 hr after treatment, revealed that TIOX-treated worms absorbed and metabolized much smaller quantities of exogenous glucose than did the controls and that the ability of the worm to accumulate glucose against a concentration difference was significantly depressed. A mode of action common to the structurally related benzothiazole and benzimidazole anthelmintics is indicated by the similarity of their biochemical and physiological effects on the tapeworms and their time course of action when administered to rats infected with H. diminuta. Molecular modeling revealed that the benzothiazole and benzimidazole anthelminitics are congruent electronically and structurally. In vivo drug efficacy depends upon the magnitude of the molecular dipole moment and the percentage of polar surface area. Within the benzimidazole series, structural and electronic congruence is found between the 2-thiazolyl and 2-methyl carbamate groups, suggesting that these groups behave similarly in transport to, and binding at, the active site. Finally, anthelmintics that have the 5' substituents twisted out-of-plane were more active than those anthelminitics with 5' substituents in-plane. All of these factors implicate a highly polar, L-shaped cleft to which the anthelmintics bind at the active site.

Photocycle-dependent emergence by cercariae of Halipegus occidualis from Helisoma anceps, with special reference to cercarial emergence patterns as adaptations for transmission.

Abstract: Emergence by cercariae of Halipegus occidualis (Hemiuridae) from naturally infected Helisoma anceps (Pulmonata) was evaluated with respect to change in temperature and light. Total cercarial emergence per snail per day increased with temperature in 2 experiments: at constant temperatures of 16, 22, and 28 C, and at temperatures varying within the range 15-30 C. The number of cercariae emerging per snail per day varied extensively among snails and from day to day for individual snails. The proportion of cercariae that emerged during darkness in each 24-hr period on a 12-hr light: 12-hr dark photocycle was consistent for each snail over 3 photocycles, but it varied among snails: a mean of 73% of cercariae emerged during darkness at 16 C, 84% at 22 C, and 89% at C. The ecological consequences of nocturnal emergence by sessile, long-lived cercariae, such as those of H. occidualis, are discussed with reference to 3 hypotheses: synchronization with activity of the next host, enhancement of dispersal, and reduction of mortality. Daily cycles of emergence by cercariae from molluscan hosts are reported widely (Rees, 1948; Macy et al., 1960; Asch, 1972; Betterton, 1979; Th6ron, 1985, 1989; Lewis et al., 1989). These cycles of emergence correspond to daily changes in ambient light or temperature and often correlate with activity cycles of the next host (Ginetsinskaya, 1968; Cable, 1972; Betterton, 1979; Th6ron, 1984, 1985; Lewis et al., 1989). An absence of daily cycles has been reported for cercariae that encyst in the external medium following emergence (Kendall and McCullough, 1951; Ginetsinskaya, 1968). Several authors (Ginetsinskaya, 1968; Cable, 1972; Th6ron, 1984) interpreted these types of contrasting observations as evidence that daily cycles of emergence evolved as adaptations for transmission, by enhancing the ability of active, short-lived cercariae to find hosts rapidly. However, not all digeneans produce such cercariae. Cercariae of Halipegus occidualis are nonmotile and long-lived (Shostak and Esch, 1990) and they are ingested by their microcrustacean second intermediate hosts. Macy et al. (1960), in a limited study, reported that cercariae of H. occidualis tend to emerge from Helisoma subcrenatum during late afternoon and night. This observation seems contrary to the usual hypothesis to explain periodicity of cercarial emergence Received 9 April 1990; revised 12 June 1990; accepted 14 June 1990. * Present address: Department of Zoology, University of Alberta, Edmonton, Alberta, Canada T6G 2E9. (i.e., a necessity to find hosts rapidly). The present study evaluated the emergence of cercariae ofH. occidualis from Helisoma anceps, primarily with respect to photocycle but also with respect to the modifying influence of temperature. Nocturnal emergence by cercariae was confirmed, and alternate hypotheses were developed to explain periodic emergence by cercariae. MATERIALS AND METHODS Helisoma anceps was collected from Charlie's Pond, an impoundment in the piedmont area of North Carolina (described in Crews and Esch [ 1986]), in October 1987 and maintained at 20-24 C under natural lighting in a 50-L aquarium containing pond water and vegetation. Lettuce was provided. After 1-2 mo snails were screened individually for infection with H. occidualis, following the methods of Goater et al. (1989). One experiment evaluated the effect of temperature. At 1030 hr on day 0, 7 naturally infected snails (shell diameter: 9.0-11.3 mm) in individual 55-mm-diameter dishes, containing 30 ml filtered pond water and a 1-cm2 piece of lettuce, were placed in a controlled environment chamber at 22 ? 1 C. A 12-hr light: 12hr dark photocycle (light commencing 0500 hr) was established in the chamber using a 25-watt incandescent light bulb suspended 25 cm above the dishes. Based on the study by Asch (1972), it was assumed that body temperatures of H. occidualis changed negligibly when illuminated. At 1030 hr daily until day 24, each snail was transferred to a new dish containing fresh water and lettuce and cercariae in the old dishes were counted (total counts if 500 cercariae). The temperature was changed by 5 C every 3-4 days (Fig. 1). Snails were killed on day 24, and sporocysts and rediae were counted. A second experiment evaluated the effect of light. At 0700 hr on day 0, 24 infected snails (shell diameter:

Current Concepts in Parasitology

Abstract: There are twelve chapters in this book. The subjects discussed include the mechanisms of eosinophilia in parasitic infections, piscine immunity to protozoan diseases, survival strategies of trichuroid nematodes, sero-diagnosis, purine and pyrimidine biosynthesis, metabolism as targets for antiparasitic chemotherapy, ecology of marine parasites, transmission patterns and evolution of nematodes as well as coevolution.

Schistosoma mansoni modulation of phagocytosis in Biomphalaria glabrata.

Abstract: Both short-term (3 hr) exposure of Biomphalaria glabrata snails (M-line and 13-16-R1) to Schistosoma mansoni (PR1) miracidia and in vitro incubation of parasite sporocysts with host hemolymph components altered host phagocytic ability. Hemocytes obtained from susceptible (M-line) snails that had been exposed to parasite miracidia for 3 hr showed reduced levels of phagocytosis of yeast cells in vitro compared to hemocytes from unexposed individuals. Incubation of whole hemolymph with sporocysts in vitro also reduced yeast phagocytosis in this susceptible strain. In contrast, resistant (13-16-R1) hemocytes showed increased levels of yeast phagocytosis after in vitro incubation with the parasite, and the opsonic properties of 13-16-R1 plasma were greater after exposure of snails to miracidia. These strain-specific effects of S. mansoni on host hemocyte phagocytosis and plasma opsonization were seen only when both plasma and hemocytes were present at the time of exposure to the parasite.

Polymorphism of Eimerian Oocysts Can be a Problem in Naturally Infected Hosts: An Example from Subterranean Rodents in Bolivia

Abstract: Since 1986, 364 tuco-tucos (Ctenomys spp.) representing 7 species were collected from 16 major collecting areas representing at least 4 distinct ecological habitats in Bolivia, South America. All were examined for coccidia, and 125 (34%) had oocysts in their feces including 84 of 236 (36%) Ctenomys boliviensis from tropical palm/savanna habitats; 1 of 3 (33%) Ctenomys conoveri from a chaco thorn forest; 3 of 7 (33%) Ctenomys frater from medium altitude grass habitats; and 6 of 8 (75%) Ctenomys lewisi and 31 of 35 (88%) Ctenomys opimus from high altitude/puna habitats. None of 3 Ctenomys leucodon (high altitude/puna) or 72 Ctenomys steinbachi (tropical palm/savanna) were passing oocysts when examined. The 5 infected host species all had oocysts of Eimeria opimi Lambert, Gardner, and Duszynski, 1988, in their feces. These oocysts and their sporocysts varied greatly in size, both within and between host species, but qualitative characters (e.g., residua and wall texture) remained constant. Our conclusion, that all oocysts seen were E. opimi, was supported by multigroup discriminant analysis of 256 individual oocysts, 30-67 selected randomly from each Ctenomys sp. Minimum polygons enclosing the centroid (= multivariate mean) and the spread of individuals for each species group (OTU) showed significant overlap in discriminant space, and Geisser classification showed a 55% miss rate of individuals being classified into the wrong OTUs. Thus, oocyst and sporocyst lengths and widths cannot be used to separate morphotypes of E. opimi from different Ctenomys spp. from different geographic regions of Bolivia.

The pathogenicity, localization, and cyst structure of echinostomatid metacercariae (Trematoda) infecting the kidneys of the frogs Rana clamitans and Rana pipiens.

Abstract: Light and scanning electron microscopy were used to examine the localization and pathogenicity of echinostomatid metacercariae infecting the kidneys of leopard frogs, Rana pipiens, and green frogs, Rana clamitans. Cysts occurred predominantly in the ventrolateral renal cortex, and at least some were confined to the lumen of the Bowman's capsules. Each vermiform metacercarial body was enclosed by a spherical cyst wall that had a uniform thickness. The wall was composed of a homogeneous material containing basic and keratinlike proteins, with sulfated acid mucopolysaccharides on the outer surface. Most cysts were enclosed by a fibrous capsule of host origin, or were surrounded by an inflammatory focus. Fibrosis was always focal, but its degree varied between individual hosts and between different cysts within the same host. Some heavily encapsulated cysts were darkened and contained disintegrating worms. In heavily infected kidneys, confluence of fibrotic or inflammatory foci resulted in the displacement of functional renal tissue. These data suggest that infection by echinostomatids may impair renal function and that the host's response affects parasite viability.

Trichinella spiralis in an agricultural ecosystem: transmission in the rat population.

Abstract: Four hundred forty-three Norway rats (Rattus norvegicus) were examined to determine their role in the transmission and maintenance of Trichinella spiralis on a pig farm. Rats, classified by sex and weight, were examined for trichinellosis by peptic digestion of muscle samples. Over a 25-mo period, 188 (42.4%) rats were found to be infected with T. spiralis. The mean intensity of infection was 293.2 larvae per gram (LPG) of muscle; 65 (34.6%) infected rats had intensities of infection greater than 100 LPG. Even in the absence of a known source of infected meat (garbage containing meat scraps or dead animals), the rat population maintained the infection, probably through cannibalism. Population reduction was an effective method for reducing the prevalence of infection within the rat population. Therefore, to reduce the likelihood of transmission of T. spiralis between rats and swine, it is essential that rat populations in a farmyard environment be controlled.

Short-tailed shrews as reservoirs of the agents of Lyme disease and human babesiosis.

Abstract: To determine whether short-tailed shrews (Blarina brevicauda) serve as reservoir hosts for the Lyme disease spirochete (Borrelia burgdorferi) and the agent of human babesiosis (Babesia microti), we examined nymphal ticks that had fed as larvae on shrews collected from 3 enzootic sites in coastal Massachusetts for evidence of infection by either or both of these agents. Xenodiagnosis indicated that 11 of 14 shrews were infected by B. burgdorferi. One of 3 piroplasm-infected shrews also infected ticks with B. microti. In a site where the piroplasm is endemic, 11 of 17 shrews showed patent parasitemias by thin blood smears. Of these, 4 had parasitemias exceeding 40%. Few immature ticks infested shrews, however, suggesting that B. brevicauda, although abundant in some endemic sites and serving as a competent reservoir, would contribute minimally to the population of infected nymphs.

Effects of sulfadiazine and amprolium on Neospora caninum (Protozoa: Apicomplexa) infections in mice.

Abstract: An immunosuppressed mouse model was used to determine the effects of amprolium and sulfadiazine on experimental Neospora caninum infections. Both drugs were given in the drinking water. Neither drug was effective in treating infections when given 7 days after inoculation of tachyzoites, when clinical signs of disease had developed. Amprolium did not prevent deaths or development of clinical signs when given in the drinking water at 1 mg/ml or 5 mg/ml 3 days after inoculation of tachyzoites. Sulfadiazine in drinking water was not effective when given at 0.5 mg/ml but was effective in preventing deaths and clinical disease when given at 1 mg/ml 3 days after inoculation with tachyzoites. Most mice (6 of 10) treated for 3 days with 1 mg/ml sulfadiazine in drinking water developed encephalitis after drug treatment was stopped. Treatment for 14 days with 1 mg/ml sulfadiazine in drinking water was needed to protect 90% of inoculated mice.

Immunohistochemical demonstration of Trypanosoma evansi in tissues of experimentally infected rats and a naturally infected water buffalo (Bubalus bubalis).

Abstract: Trypanosoma evansi was demonstrated by an immunohistochemical technique in formalin-fixed paraffin-embedded tissues of experimentally infected rats. Trypanosoma evansi was visible readily, nuclei were stained darkly, the cytoplasm was stained moderately, and the cell membranes were delineated clearly. The parasites were present in small- to large-sized blood vessels of all organs, in extravascular spaces of ventricles and neuropil of the brain, and in interstitial tissues of the lung and testes. This method also stained nuclei but not cytoplasm or cell membranes of Trypanosoma congolense, and did not stain Trypanosoma theileri. In a water buffalo (Bubalus bubalis) with nonsuppurative meningoencephalitis, the presence of T. evansi could not be demonstrated by conventional histological stains. However, the trypanosomes were recognized readily in the Virchow-Robin spaces and neuropil of the brain by the immunohistochemical method.

Tissue "cysts" of Hepatozoon griseisciuri in the grey squirrel, Sciurus carolinensis: the significance of these cysts in species of Hepatozoon.

Abstract: The lungs of a grey squirrel infected with Hepatozoon griseisciuri contained, in addition to typical haemogregarine schizonts, small cysts, each of which contained a single cystozoite. The presence of these cysts, which resemble those recorded in Hepatozoon species in reptiles, suggests that they may be a common feature in all Hepatozoon species and that these parasites may be transmitted by predation as well as by ingestion of infected arthropod vectors.

A novel anthelmintic model utilizing jirds, Meriones unguiculatus, infected with Haemonchus contortus

Abstract: Currently, no in vivo laboratory model is available for evaluating anthelmintics against the important ruminant helminth Haemonchus contortus. This report outlines a novel anthelmintic assay utilizing immunosuppressed (0.02% hydrocortisone in feed) jirds, Meriones unguiculatus, infected with H. contortus. Immunosuppressed jirds were inoculated with approximately 1,000 exsheathed infective larvae of H. contortus, treated per os on day 10 postinoculation (PI), and necropsied on day 13 PI. Each stomach was removed, opened longitudinally, incubated in distilled water at 37 C for 5 hr, fixed in formaldehyde solution, and stored for subsequent examination. Stomach contents were examined using a stereomicroscope (15-45x). A variety of standard anthelmintics has been evaluated in the model; modern broad-spectrum ruminant anthelmintics (benzimidazoles, febantel, ivermectin, levamisole hydrochloride, and milbemycin D) are active uniformly and in most cases at doses (mg/kg) comparable to those required for efficacy against H. contortus in ruminants. This model provides an important new tool to assess preliminarily the activity of experimental drugs against H. contortus in vivo prior to studies in ruminants and also may provide a useful tool for studying host-parasite interactions for H. contortus.

Lernaeocera branchialis: a potential pathogen to cod ranching.

Abstract: A study was carried out to determine the effect of Lernaeocera branchialis on Atlantic cod infected in the laboratory and in the field and also to ascertain its effect on cod-ranching. Sixty-four percent (308) of 481 cod acquired infections in the laboratory and 33% (159) of the infected fish died over a 4-yr period. About 74% of the deaths occurred within 4 mo of the infection. Monthly samples of cod collected adjacent to a cod-ranching operation showed an initial prevalence of 30% that subsequently decreased in the following 2 mo to 15%. Prevalence of the infection also decreased among the initial field sample of cod that were kept alive, from 30 to 17% during the same 2-mo period and to 9% after 8 mo and was associated with death caused by the parasite. Cod examined at intervals after infection showed evidence of reduced weight gain, lower liver somatic index, liver lipid, and blood values than controls. A field sample taken from the same area during the summer of the following year indicated a prevalence of 12%. This higher than usual prevalence (4-6%) was associated with retention of the intermediate host, Cyclopterus lumpus, that provided an additional source of infective stages. It is likely that increased parasitism could affect the success of the cod-ranching operation in view of the parasite's devastating effects on its hosts.