S. S. Johnson

A coprological survey of parasites of wild neotropical felidae.

In conjunction with an ecological study of jaguars in the Cockscomb Basin of Belize, Central America, fecal samples from jaguars (Panthera onca), jaguarundis (Felis yagouaroundi), ocelots (Felix pardalis), and pumas (Felix concolor) were examined for parasite products (eggs, larvae, and oocysts). Of the 45 samples examined, 39 (86.7%) were positive for parasite products, 23 of 25 (92%) jaguar samples were positive, as were all of the puma (4/4) and ocelot (8/8) samples. Four of 6 samples from unknown species were positive (66.7%). Two jaguarundis samples were negative. The following were identified in the samples: Paragonimus sp. eggs, Taeniidae eggs, Strongylate eggs, Toxocara cati eggs, Toxascaris sp. eggs, Capillaria sp. eggs, Spiruridae eggs, Aelurostrongylus sp. larvae, Oncicola sp. eggs, Hammondia pardalis oocysts, Isospora sp. oocysts, Toxoplasma gondii-like oocysts and Sarcocystis sp. sporocyst.

Prehistoric parasitism in Tennessee: evidence from the analysis of desiccated fecal material collected from Big Bone Cave, Van Buren County, Tennessee.

Eight samples of desiccated human feces collected from Big Bone Cave (40VB103), Van Buren County, Tennessee, were analyzed to determine the presence of ecto- and endoparasitic infection among the prehistoric population using the cave. Radiocarbon-dated torch material from the cave indicated that it was a locus of human activity 2,177 +/- 145 yr ago. Parasitic species identified were: Ascaris lumbricoides, Enterobius vermicularis, fleas of the tribe Phalacropsyllini, and protozoan cysts. The cysts were identified as Giardia using an indirect immunofluorescent antibody test. The only report of Giardia in a prehistoric context is the identification of cysts in 2 1,800-yr-old paleofecal specimens from a cave in Israel. This is the first report of Giardia from paleofeces in the New World.

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

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.

VIABILITY OF INFECTIVE LARVAE OF HAEMONCHUS CONTORTUS, OSTERTAGIA OSTERTAGI, AND TRICHOSTRONGYLUS COLUBRIFORMIS FOLLOWING EXSHEATHMENT BY VARIOUS TECHNIQUES

Various techniques were examined to determine optimum conditions for exsheathing infective larvae of 3 important ruminant parasites (Haemonchus contortus, Ostertagia ostertagi, and Trichostrongylus colubriformis). In repeated experiments, aliquots of 10(5)-10(6) infective larvae, 1-2 mo old, of each parasite were incubated in each of 4 exsheathing media (distilled water, Earles balanced salt solution + carbon dioxide, nematode washing buffer + carbon dioxide, or sodium hypochlorite) for 1 or 18 hr. In each case, the percentage of larvae exsheathed and infectivity for jirds was determined. Results of these studies indicate that no single exsheathing technique of those studied is optimum for every parasite. In addition, caution must be used in drawing conclusions from in vitro studies using exsheathed larvae because techniques that routinely provide high percentages of exsheathment also appear to reduce viability.

Concurrent infections with the ruminant nematodes Haemonchus contortus and Trichostrongylus colubriformis in jirds, Meriones unguiculatus, and use of this model for anthelmintic studies.

Haemonchus contortus- and Trichostrongylus colubriformis-infected jirds (Meriones unguiculatus) are useful for anthelmintic studies. With concurrent infections of these parasites established in the jird, questions of not only anthelmintic activity, but to some extent spectrum, could be assessed in a single model system. This report outlines a model using immunosuppressed (0.02% hydrocortisone in feed) jirds concurrently infected with H. contortus and T. colubriformis. Immunosuppressed jirds were inoculated with approximately 1,000 exsheathed infective larvae of each species, treated per os on day 10 postinoculation (PI), and killed on day 13 PI. Stomachs and small intestines were removed, opened longitudinally, incubated in distilled water at 37 C for 5 hr, fixed in formaldehyde solution, and stored for subsequent examination. Contents of both organs were examined using a stereomicroscope (15-45 x). Various standard anthelmintics were 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 comparable to those required for efficacy against these parasites in ruminants. This model, using worms of 2 genera living in distinct sites, allows preliminary evaluation of anthelmintic activity and spectrum for experimental compounds in a single cost- and resource-efficient experiment.

PREVALENCE OF TOXOPLASMA GONDII ANTIBODIES IN NINE POPULATIONS OF DAIRY GOATS: COMPARED TITERS USING MODIFIED DIRECT AGGLUTINATION AND INDIRECT HEMAGGLUTINATION

In a convenience survey of dairy goats on 9 farms in Tennessee, Kentucky, and Georgia, 54 of 99 females were positive for antibodies to Toxoplasma gondii as measured by the indirect hemagglutination test (IHA). Two of 9 males were also positive. Positive goats were found on all farms. The percentage of positive does increased from 55% to 65% when the sera were titered a second time by modified direct agglutination (MDAT). This difference was not statistically significant, indicating that both IHA and MDAT are reliable epidemiological tools.

Growth and development of Haemonchus contortus in jirds, Meriones unguiculatus

Growth and development of Haemonchus contortus were examined in jirds and were compared to these processes in lambs. Number, sex, size, and stage of development were determined for worms recovered at necropsy at various times postinoculation (PI) from immunosuppressed jirds inoculated with approximately 1,000 exsheathed infective larvae (L3) of H. contortus. In addition, gastric tissue samples from jirds were examined histologically. Parallel studies were done in lambs inoculated with approximately 7,500 L3. Typically, 5-30% of the inoculum established and survived in jirds at reasonably stable numbers to day 14 PI. By day 21 PI, worm numbers in jirds decreased dramatically. Although the parasite was similar in size and development on day 4 PI in jirds and lambs, from day 7 PI on, worms were significantly smaller and less developed in jirds. On histological examination, the parasite was found only in the glandular portion of the stomach of jirds (anatomically similar to its predilection site in the abomasum of lambs), and histological changes were consistent for both host species. Although growth and development of H. contortus are slower and incomplete in jirds, the parasite establishes, grows, and develops (at anatomically comparable sites in both hosts) in this model. Thus, the model appears to provide a useful laboratory host to study H. contortus.

UTILITY OF A HAEMONCHUS CONTORTUS/JIRD (MERIONES UNGUICULATUS) MODEL FOR STUDYING RESISTANCE TO LEVAMISOLE

Trichostrongylid nematodes of sheep commonly are identified as exhibiting resistance to levamisole. In vitro assays have been developed to study levamisole resistance for Haemonchus contortus, but no in vivo model has been identified for this species. To determine the utility of a H. contortus/jird (Meriones unguiculatus) model for examining levamisole resistance, immunosuppressed jirds were inoculated with approximately 1,000 exsheathed infective larvae of H. contortus (resistant or susceptible to levamisole), treated per os on day 10 postinoculation (PI) with levamisole hydrochloride or analogs of the drug, and killed on day 13 PI. Stomachs were removed, opened longitudinally, incubated in distilled water at 37 C for 5 hr, fixed in formaldehyde solution, and stored for subsequent microscopic examination. Doses of levamisole and its analogs, which elicited percentage clearances of greater than or equal to 93.5 for the susceptible strain, cleared less than or equal to 68.9% of the resistant worms. These data are consistent with activities for the drugs against wild-type and levamisole-resistant strains of Caenorhabditis elegans. Thus, the H. contortus/jird model provides a useful in vivo tool to study resistance to levamisole and possibly other anthelmintics.

INFECTIVITY OF HYMENOLEPIS DIMINUTA FOR THE JIRD, MERIONES UNGUICULATUS, AND UTILITY OF THIS MODEL FOR ANTHELMINTIC STUDIES

The jird (Meriones unguiculatus) has been shown to be a useful model host for the cestodes Taenia crassiceps and Echinococcus multilocularis. This report outlines a novel model in which hydrocortisone-treated jirds (0.02% in the feed) are infected with another cestode, Hymenolepis diminuta. Jirds were inoculated with 5 freshly harvested cysticercoids of H. diminuta prior to (day 0, -1, or -5) or after (day 1 or 5) switching to medicated feed; in some cases, jirds were never medicated. On days 7, 14, 21, or 28 postinoculation (PI), jirds were killed by CO2 inhalation and their small intestines were examined for tapeworms. Hymenolepis diminuta established, grew, and developed to the gravid adult state in jirds. They persisted longer in medicated (21 days) than in nonmedicated (7 days) animals, and generally higher levels of infection were obtained when jirds were inoculated immediately prior to switching to medicated feed. Treatment of infected jirds on day 4 or days 4, 5, and 6 PI with selected anthelmintics followed by necropsy on day 7 PI discriminated drugs with known activity against tapeworms from those with little or no activity. This rodent in vivo model should provide a useful adjunct for anthelmintic studies.

Kinetics of expulsion of Haemonchus contortus from sheep and jirds after treatment with closantel

Experiments were conducted in sheep after intramuscular treatment with closantel and in jirds after oral treatment with closantel to determine when expulsion of established H. contortus commences. Expulsion starts at about 8 h in sheep and coincides with the onset of reduced motility in worms recovered from the abomasum. In jirds, expulsion starts by 2 h after treatment. Experiments also conducted in jirds showed that infective larvae are first killed by circulating closantel 3 days after infection, when blood feeding starts, and that by 8 days 80% of larvae are lost.