Evaluation of the efficacy of emodepside+praziquantel topical solution against cestode (Dipylidium caninum, Taenia taeniaeformis, and Echinococcus multilocularis) infections in cats.

Emodepside+praziquantel topical solution was developed to provide broad-spectrum anthelmintic activity against gastrointestinal parasites in cats. Eight controlled studies were conducted to evaluate the efficacy of a topical solution of emodepside (3 mg/kg) and praziquantel (12 mg/kg) (Profender, BayerAG, Leverkusen, Germany) against feline infections with three species of cestodes. Studies featured naturally acquired infections of Dipylidium caninum or Taenia taeniaeformis, or experimental infections with Echinococcus multilocularis that were placebo-controlled, randomized and blinded. Cats were euthanatized and necropsied between 2 and 11 days after treatment, depending on the target parasite. The efficacy of emodepside+praziquantel topical solution was 100% against D. caninum and T. taeniaeformis, and 98.5- 100% against E. multilocularis. No significant systemic or local adverse reactions to treatment were noted in cats that received the combination. Topical treatment of cats with emodepside+praziquantel topical solution was safe and highly effective against cestode infections.

Raccoon ascarid larvae (Baylisascaris procyonis) as a cause of ocular larva migrans.

Larvae of the common raccoon roundworm, Baylisascaris procyonis, are known causes of visceral larva migrans and CNS disease in animals and human beings. In the present experiments we examined the ability of B. procyonis to cause ocular larva migrans (OLM) in subhuman primates, as an indication of its possible ocular zoonotic importance. Squirrel monkeys given 5,000 or 10,000 infective B. procyonis eggs per os and cynomolgus monkeys given 20,000 eggs had clinical and histologic evidence of OLM, beginning 7 days after inoculation. Clinically, multifocal retinal hemorrhages, white spots, chorioretinitis, inflammatory tracks, vascular sheathing, diffuse retinal degeneration, and motile intraretinal larvae were seen. Histologically, primarily subretinal larvae caused varying degrees of retinal disruption, degeneration and necrosis, retinitis, vasculitis, and perivascular sheathing, primarily with eosinophils. Larvae were also present in choroidal granulomas. It was concluded that B. procyonis larvae have marked ability to produce OLM in subhuman primates following oral infection and should be considered as a possible etiology in human ocular disease.

Characterization of the clinical and anatomical pathological changes associated with Hepatozoon mocassini infections in unnatural reptilian hosts.

Laboratory-reared Aedes aegypti mosquitoes were employed in the successful transmission of Hepatozoon mocassini from a cotton-mouth moccasin (Agkistrodon piscivorus leucostoma) to 3 lizard species (Sceloporus undulatus, Eumeces obsoletus and Sceloporus poinsetti). Marked to severe lethargy and anorexia developed in the S. undulatus, E. obsoletus and S. poinsetti at 15, 38, and 96 days postinfection (PI), respectively. All 3 lizards developed a leukocytosis and had increased plasma aspartate aminotransferase activity (AST) by 14 days PI. Multifocal random hepatocellular necrosis and intrahepatic aggregates of heterophils centered on mature H. mocassini meronts were demonstrated in all 3 lizards. The pulmonary interstitium was multifocally thickened by aggregates of heterophils centered on meronts. No comparable clinical or anatomical pathological changes were demonstrated in naturally infected snakes. The results of this study suggest that H. mocassini is capable of inducing necrotizing inflammatory by lesions in unnatural reptilian hosts.

Loaiasis (Loa loa) in an African student in Indiana.

Loaiasis was seen in an African student in Indiana, who had non-specific complaints, leukocyte count of 8,400/mm3, a 47% eosinophilia, and a high Loa loa microfilaremia. Successful treatment with diethylcarbamazine (Hetrazan) produced no allergic manifestations and no reactions to the adult parasites were evident.

Immunofluorescent localization of eosinophil granule major basic protein in fatal human cases of Baylisascaris procyonis infection.

We examined eosinophil degranulation in tissues from patients infected with Baylisascaris procyonis as shown by the extracellular deposition of granule major basic protein (MBP). We utilized immunofluorescence to localize MBP in eosinophils and at sites of degranulation to study specimens from 2 fatal cases of B. procyonis infection. Large numbers of intact eosinophils were present in the brain around blood vessels and necrotic migration tracks and in mesenteric granulomata. Extensive extracellular MBP deposition was present in the necrotic migration tracks in the brain and around larvae in the mesenteric granulomata in association with the radiating eosinophilic deposits characteristic of the "Splendore-Hoeppli phenomenon." The Splendore-Hoeppli deposits consist in part of eosinophil granule MBP. Release of the cytotoxic MBP in response to invading larvae may cause tissue damage. Central nervous system tissue damage by cytotoxic eosinophil granule proteins may contribute to the neurologic symptoms of B. procyonis infection.

Two cases of intraocular infection with Alaria mesocercaria (Trematoda)

We encountered two cases of human intraocular infection with mesocercariae of Alaria (Trematoda), involving unrelated Asian men who had unilateral decreased vision. Both patients had pigmentary tracks in the retina, areas of active or healed retinitis, or both, and other signs of diffuse unilateral subacute neuroretinitis. Similar, nonnematode worms were seen in the patients' retinas and vitreous, respectively, several years after apparent infection. The worm in Case 1 was analyzed from projected fundus photographs and diagnosed as an Alaria mesocercaria on the basis of its shape, size (500 x 150 microns), and movement; it was successfully killed with laser. The worm in Case 2 was removed surgically from the vitreous and identified as A. mesocercaria, 555 x 190 microns, most likely A. americana. The probable source of infection in the patients was ingestion, in local restaurants, of undercooked frogs' legs containing A. mesocercaria. In addition to causing prolonged intraocular infection, A. mesocercaria was found to be a cause of diffuse unilateral subacute neuroretinitis, a condition previously attributed only to intraocular nematode larvae.

Radiologic-pathologic findings in raccoon roundworm (Baylisascaris procyonis) encephalitis.

A 13-month-old boy developed eosinophilic meningoencephalitis, retinitis, and a protracted encephalopathy with severe residual deficits. The initial MR examination revealed diffuse periventricular white matter disease, and follow-up images showed atrophy. Brain biopsy, serology, and epidemiologic studies lead to the diagnosis of Baylisascaris procyonis infection, a parasitic disease contracted through exposure to soil contaminated by the eggs of a common raccoon intestinal roundworm. The pathologic, epidemiologic, and imaging features of this disease are herein reviewed.

A tongue biopsy technique for the detection of trichinosis in swine.

A tongue biopsy technique developed for the detection of Trichinella spiralis infection in swine involves taking a deep core biopsy of the tongue musculature, and examination of the sample by digestion. Using this procedure, 31 of 52 (60%) swine from an Indiana herd were found to be infected with T. spiralis. The average biopsy weighed 0.42 g, and the intensity of infection averaged 180 larvae per gram (range 2-1157). The biopsy was quick and easy to perform and the tongues healed well following the procedure. This technique may have applicability for Trichinella detection in epidemiological, control and research studies on swine and other animals.

Experimental cerebrospinal nematodiasis due to Baylisascaris procyonis in chickens.

Avian cerebrospinal nematodiasis was studied in chickens experimentally inoculated with infective eggs of Baylisascaris procyonis of raccoon origin. Birds each received 3,200, 1,600, 800, 400, 200, or 0 eggs, one dose per group of 10 birds. Central nervous system (CNS) disease developed in 17 of the 50 infected birds. Birds receiving the higher dosages exhibited the worst clinical signs, had higher mortality rates, lived for shorter duration, and had the most severe brain lesions. Onset of CNS disease was consistent between groups and averaged 20.4 days, and duration varied from less than one to 23 days. No gross lesions were present. Histologically, multifocal areas of malacia and necrosis, hemorrhage, perivascular cuffing, nonsuppurative meningoencephalitis, focal granulomas, and migrating larvae were seen in the brains. No larvae were recovered from the brains by the Baermann method. Extraneural lesions were limited to focal choroiditis and a granuloma in an extrinsic ocular muscle.

Atypical histomoniasis in Bobwhite quail.

An outbreak of histomonad infection of an apparent atypical nature was diagnosed in a flock of about 850 Bobwhite quail. Mortality was 95% over a 3-week period. The most prominent gross pathologic lesions were in the livers: disseminated white foci of necrosis, 1 to 2 mm in diameter, and subcapsular multifocal splenic necrosis was seen occasionally; lower intestinal lesions were infrequent. Histologic examination of liver and spleen sections revealed focal necrosis associated with variable numbers of protozoal organisms identified as a Histomonad spp. Identification of the protoza was ascertained by electron microscopy. Histmonads were isolated from affected quail livers and propagated in specific-pathogen-free chicken embryos. Lesions produced in embryos were evaluated. Isolates of the organism were used to reproduce the disease in young Bobwhite quail.

Verminous encephalitis in commercial chickens.

An outbreak of fatal encephalitis in four-week-old chickens is described. The causal agent was shown to be Baylisascaris procyonis, the common roundworm of raccoons.

Fatal hepatic trematodiasis in cockatoos due to Platynosomum proxillicens.

Severe hepatic trematodiasis due to Platynosomum proxillicens was diagnosed as the cause of death in two sulfur-crested cockatoos (Cacatua sulfurea). Gross pathologic alterations included hepatomegaly in one bird and hepatic and generalized cyanosis in the other. Microscopic changes included multifocal hepatic necrosis, dilated fibrous bile ducts containing trematode, parasites, bile duct hyperplasia, and hepatic inflammation.

Echinococcus multilocularis identified in Indiana, Ohio, and east-central Illinois.

Echinococcus multilocularis was identified in a coyote in Indiana in January 1990. Subsequently, 300 wild canids (red foxes, gray foxes, coyotes) were collected from Indiana, Ohio, Illinois, Michigan, and Kentucky and examined for this parasite. Of these, 41 (13.7%) were infected with E. multilocularis. Infected animals were from northern and central Indiana, northwestern Ohio, and east-central Illinois. Twenty-nine of 162 (17.9%) animals from northern and central Indiana were positive, versus 0 of 83 from southern Indiana. Infected animals from northern and central Indiana included 16 of 71 (22.5%) red foxes and 13 of 70 (18.6%) coyotes. In Ohio, 6 of 22 (27.3%) red foxes were infected, and in Illinois, 6 of 17 (35.3%) coyotes were positive. Mean intensity of infection for red foxes was 372 worms (range, 2-3, 640), and for coyotes, 6,579 worms (range, 1-52,000). These findings constitute new state and distribution records for E. multilocularis and a significant range extension for this parasite species in central North America.

Susceptibility of Peromyscus leucopus and Mus musculus to infection with Baylisascaris procyonis.

In this study, we compared the susceptibility of Peromyscus leucopus (white-footed mouse), a common natural intermediate host, and Mus musculus, a commonly used experimental model, to infection with larvae of the raccoon ascarid, Baylisascaris procyonis. Three groups of 10 mice of each species were given 50, 250, or 500 infective B. procyonis eggs by gavage. The mice were observed daily for clinical signs of central nervous system (CNS) disease and at necropsy the distribution of larvae in 10 body regions and organs was determined and compared. Clinical CNS disease developed in 57% of P. leucopus and 93% of M. musculus. The average clinical incubation period was significantly longer in P. leucopus (20.6 days postinfection [PI]) than in M. musculus (10.7 days PI), and clinical disease progressed slower in P. leucopus. Significantly fewer larvae were recovered from P. leucopus than from M. musculus. Most larvae were recovered from the anterior carcass and viscera of P. leucopus and from the carcass, head, and brain of M. musculus. CNS invasion was dose dependent in M. musculus but not in P. leucopus. Few or no grossly visible larval granulomas were present in P. leucopus but were abundant in M. musculus. We concluded that P. leucopus was less susceptible than M. musculus to B. procyonis infection, based on a decreased intensity of infection, longer clinical incubation period or lack of clinical disease, slower progression of disease, different larval distribution, and lower tissue reactivity to larvae.

Scanning electron microscopy of the labia of Baylisascaris procyonis (Nematoda).

The labial organization of adult Baylisascaris procyonis was studied by scanning electron microscopy, and found to be similar for males and females. The apical part of each lip was smooth, and the basal part reticulated. The dorsal lip possessed two dorsolateral double papillae and two internal labial papillae; the two subventral lips each had one ventrolateral double and one externolateral papilla, two internal labial papillae, and an amphid. The small papilla of each double set was dome-shaped and smooth, whereas the large papilla was broad and had a prominent central pore. The externolateral papillae had raised, highly sculptured surfaces with numerous slits and creases present. Amphids resembled those of previously studied nematodes. The internal labial papillae consisted of pits. Denticles arose as a single row from the apical edge of the inner labial surface, were usually evenly spaced, and pyramidal or conoidal in shape. They were typically unicuspid, but bicuspid denticles were occasionally seen. Denticle shape and size varied between specimens and on each specimen. A pit was seen in the cuticle in the central region of the denticular row of all lips. Several of these findings represent new information concerning ascaridoid nematodes.

Molecular characterization of sylvatic isolates of Trichinella spiralis.

Genetic relationships of 20 Trichinella isolates from Indiana wildlife were assessed and compared to Trichinella isolated from an infected swine herd. Trichinella larvae were isolated from coyotes, mink, raccoons, and red foxes. The larvae were maintained and amplified in white mice (ICR) and wild mice (Peromyscus leucopus). Differences in phenotypic characters of sylvatic isolates in the 2 laboratory hosts included an approximately 10-30-fold increase in parasite fecundity in wild mice. DNA for each isolate was extracted from Trichinella larvae and analyzed by dot-blot hybridization using a repetitive DNA probe pBP2 that recognizes DNA sequences specific for swine Trichinella. The probe hybridized only to Trichinella from swine and a single coyote isolate. Restriction endonucleases were used to digest DNA and the resulting fragments were separated by gel electrophoresis. Based on the presence of repetitive DNA sequences in the Trichinella genome, distinctive banding patterns were seen among the isolates. Trichinella isolated from swine had a pattern distinct from all sylvatic isolates except 1 from a coyote. Because this coyote was from the same general locality as the swine Trichinella outbreak, it was concluded that the isolate represents transmission of swine trichinellosis to the wildlife population. Further analysis using the enzyme Cla I identified unique banding patterns for wild isolates, suggesting that the sylvatic group is a genetically heterogeneous complex.

Distribution and prevalence of Echinococcus multilocularis in wild predators in Nebraska, Kansas, and Wyoming.

To further determine the distribution and prevalence of Echinococcus multilocularis in the central United States, 245 wild canids (125 red foxes, 120 coyotes) and 33 bobcats were collected from Nebraska, Kansas, and Wyoming and examined for this parasite. Animals examined included 11 red foxes from the western panhandle of Nebraska; 5 red foxes and 30 coyotes from southern Nebraska; 56 red foxes and 1 coyote from northeastern Nebraska; 20 red foxes, 63 coyotes, and 13 bobcats from northern Kansas; 2 red foxes, 26 coyotes, and 20 bobcats from southern Kansas; and 31 red foxes from east-central Wyoming. Of these, 27 of 72 (37.5%) red foxes from Nebraska were positive, including 2 of 11 (18.2%) from the western panhandle and 25 of 56 (44.6%) from the northeastern part of the state. Mean intensity of infection was 282 worms (range, 1-5,150). New distribution records were established for E. multilocularis in western Nebraska as well as for several northeastern counties. These findings support previous estimates that the southernmost front of the parasite's range extends along the southern border of Wyoming, eastward through central Nebraska and central Illinois into Indiana and Ohio.

Physicochemical characterization and monoclonal and polyclonal antibody recognition of Baylisascaris procyonis larval excretory-secretory antigens.

Baylisascaris procyonis larval excretory-secretory (ES) antigens consisted of complex glycoproteins ranging from 10 kDa to over 200 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and lectin binding. Five monoclonal antibodies (Bapr1-Bapr5) produced against B. procyonis ES antigens were assayed by western blotting with larval ES antigens from B. procyonis, Baylisascaris melis, Baylisascaris transfuga, Ascaris suum, and Toxocara canis. Bapr1 and Bapr2 recognized periodate-sensitive epitopes on 14-kDa ES components of B. procyonis, B. melis, and B. transfuga, whereas Bapr4 and Bapr5 recognized periodate-resistant epitopes present on 55-kDa ES components of B. procyonis and B. melis. Bapr3 primarily recognized periodate-resistant epitopes on 33-45-kDa components of B. procyonis and B. melis ES. Heterologous rabbit antisera cross-reacted with many B. procyonis ES antigens on western blots, but recognition of the 33-45-kDa components was genus-specific. Normal human sera and T. canis-positive human sera also cross-reacted with many B. procyonis ES antigens, including those of 33-45 kDa. However, periodate oxidation markedly decreased cross-reactions and allowed for differential immunodiagnosis of B. procyonis versus T. canis. These studies demonstrated that antibody recognition of carbohydrate epitopes on ES components is an important cause of cross-reactions in antibody detection assays. Recognition of periodate-resistant (protein) epitopes on the 33-45-kDa B. procyonis ES components appears to be useful for genus-specific immunodiagnosis of larva migrans caused by Baylisascaris spp.

Toxoplasma gondii antibodies in naturally exposed wild coyotes, red foxes, and gray foxes and serologic diagnosis of Toxoplasmosis in red foxes fed T. gondii oocysts and tissue cysts.

Antibodies to Toxoplasma gondii were determined in sera from 222 coyotes (Canis latrans), 283 red foxes (Vulpes vulpes), and 97 gray foxes (Urocyon cinereoargenteus) from Indiana, Kentucky, Michigan, and Ohio during 1990-1993. Sera were examined in 1:25, 1:100, and 1:500 dilutions by the modified direct agglutination test (MAT) with formalinized whole tachyzoites plus mercaptoethanol. Antibodies were found in 131 (59.0%) of 222 coyotes, 243 (85.9%) of 283 red foxes, and 73 (75.3%) of 97 gray foxes. Antibodies were also measured by different serologic tests in 4 littermate T. gondii-free red foxes fed T. gondii tissue cysts or oocysts; the fifth littermate fox was not fed T. gondii. Antibodies were measured in fox sera obtained 0, 14, and 36-55 days after infection with T. gondii. All 4 foxes fed T. gondii developed MAT and dye test antibody titers of 1:200 or more 14 days later. The latex agglutination test (LAT) and indirect hemagglutination test (IHAT) were less sensitive than MAT for the diagnosis of T. gondii infection in foxes. Antibodies were not detected by LAT (titer 1:64) in the 2 foxes fed tissue cysts nor by IHAT in 1 of the foxes fed tissue cysts. Toxoplasma gondii was isolated by bioassay in mice from tissues of all 4 foxes fed T. gondii. The control fox had no T. gondii antibodies detectable by any of the serologic tests.

Improved method for recovering ascarid and other helminth eggs from soil associated with epizootics and during survey studies.

Two noncentrifugal and 5 centrifugal flotation methods for the recovery of infective Baylisascaris procyonis eggs from contaminated soil were evaluated after an epizootic of fatal cerebrospinal nematodiasis in bobwhites. All involved the use of inexpensive, readily available materials and routine procedures. In all but 1 of the methods, sizes of the soil samples were 30 g; in 1 centrifugal method, the sample size was 1 g. The centrifugal methods included various combinations of (i) anionic detergent (Tween) pretreatment of the soil, (ii) 3 X washing of the sediment, (iii) NaNO3 or ZnSO4 flotation solution, and (iv) resuspension and reexamination of the sediment. By the 30-g sample centrifugal methods, significantly more eggs (av 251 to 448) were recovered than by the noncentrifugal methods (av 7 and 12) or the 1-g sample method (av 44). In the most efficacious method, the combination of Tween pretreatment, washed sediment, NaNO3 flotation solution, and resuspension/reexamination of the sediment was used. This method was tested against the 1-g sample method on soil from a naturally occurring infection of Toxocara cati visceral larva migrans in a child. On initial examination of 5 areas, 3 to 10 eggs were recovered by the 30-g sample method vs 0 eggs by the 1-g sample method. Using sandbox soil and 10 replicates each, recoveries by the 30-g sample method averaged 9.2 eggs/replicate (range 5 to 12) and those by the 1-g sample method, 0.1 eggs (range 0 to 1).