Distribution of fasciolosis in Kansas, with results of experimental snail susceptibility studies.

A total of 278 veterinarians throughout Kansas were sent mail-in survey forms asking specific questions relating to their experience with fasciolosis in their practice area. Replies were received from 178 (64%) veterinarians representing six practice types; one-third reported having seen cases of fasciolosis in their practice. The results of our survey indicate that the majority of the cattle diagnosed with liver fluke disease in Kansas are imported from other areas of the USA. However, in both central and southeastern regions of Kansas, some cattle that had never been out of the state were infected with Fasciola hepatica. Thus, these areas of Kansas should be considered endemic for liver fluke disease. Methods of diagnosis, types of operations, and improvements seen after treatment were also discussed. In order to ascertain the existence of one or more possible snail intermediate hosts within Kansas, five species of lymnaeid snails were collected from central and southeastern parts of the state and tested for their susceptibility to infection by Fasciola hepatica. The snails collected included Pseudosuccinea columella, Fossaria obrussa, Fossaria bulimoides, Fossaria parva and Fossaria dalli. Of these, Pseudosuccinea columella and Fossaria bulimoides proved susceptible to experimental infection by Fasciola hepatica. Metacercariae obtained from experimentally infected snails were used to infect both a weanling calf thereby completing the life cycle of the parasite. This report is the first to identify the existence of suitable snail intermediate hosts for Fasciola hepatica in Kansas.

Strategic use of ivermectin during pregnancy to control toxocara canis in greyhound puppies.

Twenty-one greyhound bitches were bred (Day 0) and housed throughout their pregnancies on three greyhound breeding farms in Kansas. These dogs were assigned randomly to one of four treatment groups. Group A dogs (6) were given ivermectin subcutaneously (300 microg/kg) on Day 0 (the first day the dogs were bred), and Days 30 and 60 of gestation. Group B dogs (6) were given ivermectin (300 microg/kg SQ) on Day 42. Group C dogs (3) were given ivermectin (300 microg/kg SQ) on Days 0, 30, and 60 plus 10 days after whelping. Group D dogs (6) served as controls and received no anthelmintic. Bitches and puppies were moved to the university on the day after birth and were maintained inside for 28 days. Weekly quantitative fecal exams were done on the bitches during this time. The puppies were euthanized humanely at 28 days of age. Intestinal parasites were recovered, identified, counted, sexed, and preserved in either 10% formalin or frozen at -70 degrees C. The geometric mean numbers of adult Toxocara canis in the small intestines for Group A puppies (n = 40) were 2.8, 8.5 for Group B puppies (n = 39), and 29.7 for Group D puppies (n = 28). No adults were found in the Group C puppies (n = 15). The geometric mean eggs per gram of feces from the pups in group A, B, and D were 1.3, 704, and 27, 134, respectively. No eggs were recovered from the Group C pups. The strategic use of ivermectin at 300 microg/kg in greyhound bitches on Days 0, 30, and 60 of gestation reduced the worm burden carried by the puppies by 90% and the actual number of eggs passed into the environment by 99.8%. The same dose on day 42 reduced the worm burden by 71.4% and the number of eggs passed into the environment by 97.4%. This dose given on days 0, 30, and 60 plus 10 days postwhelping, reduced the worm burden by 100%, and no eggs were passed into the environment.

Efficacy of fenbendazole granules and pyrantel pamoate suspension against Toxocara canis in greyhounds housed in contaminated runs.

The efficacy of fenbendazole granules against Toxocara canis in naturally infected greyhounds housed in contaminated environments was evaluated. Eight pens, each containing three to seven greyhounds, 3-12 months of age, were randomly allotted into two treatment groups. Greyhounds in Group 1 were treated with fenbendazole granules mixed in their feed at 50 mg/kg/day for 3 consecutive days once a month for 4 months. Greyhounds in Group 2 were treated with pyrantel pamoate suspension at 5.0 mg/kg per os once a month for 4 months. Quantitative fecal examinations were performed on days 0, 10 and then on the first day of each monthly treatment. Greyhounds administered fenbendazole had fecal egg count reductions (FECRs) of 95.8 and 99.8% at 10 and 31 days following initial treatment, respectively. Greyhounds administered pyrantel pamoate had FECRs of 85.8 and 88.3% at 10 and 31 days after the first treatment, respectively. T. canis fecal egg counts conducted from Day 31 through Day 128 were significant lower in those greyhounds administered fenbendazole as compared to greyhounds administered pyrantel pamoate. Fenbendazole produced FECRs in greyhounds from Day 31 through Day 128 by 96.8-99.8%. Pyrantel pamoate reduced fecal egg counts during the same time period 71.4-98.3%.

Production and characterization of monoclonal antibodies against excretory-secretory products of Fasciola hepatica.

Four monoclonal antibodies (mAbs) (9.49, 24.27, 46.71 and 179.57) were produced against Fasciola hepatica excretory-secretory products. Isotype analysis revealed the antibodies to be IgM, IgG3, IgG1, and IgM. In immunoblot assays, the mAbs recognized different antigenic polypeptides migrating between 29 and 180 kDa. Specificity of the mAbs was evaluated by ELISA against antigens of Fascioloides magna, Anoplocephala magna, Stichorchis subtriquetrus, Haemonchus contortus, sheep liver extract (SLE), bovine liver extract (BLE), bovine serum albumin (BSA), bovine viral diarrhea virus (BVDV), and Madin-Darby bovine kidney (MDBK) cells. Monoclonals 9.49 and 24.27 were specific, and reacted only with Fasciola hepatica antigens. However, mAb 46.71 cross-reacted with antigens of Fascioloides magna, A. magna, Stichorchis subtriquetrus, and H. contortus but not with SLE, BLE, BSA, BVDV or MDBK cells. Monoclonal antibody 179.57 cross-reacted with Fascioloides magna, A. magna, S. subtriquetrus, H. contortus, SLE, and BLE, but not with BSA, BVDV, or MDBK cells.

The hispid cotton rat (Sigmodon hispidus) as an experimental host for the bovine liver fluke (Fasciola hepatica).

The results of a study testing the susceptibility of the hispid cotton rat, Sigmodon hispidus, to infection with Fasciola hepatica are described. Ten cotton rats were placed into one of four test groups, one uninfected control or three animals each receiving two, five, or 10 F. hepatica metacercariae. Three of nine (33%) became infected, and two of these animals carried the infection to patency. At necropsy one animal had two mature F. hepatica within the common bile duct, and the other harbored a single fluke. Gross and microscopic changes, primarily of the liver, associated with infection are described, as are fecal egg counts throughout the duration of infection.

Effect of 0.29% w/w fipronil spray on adult flea mortality and egg production of three different cat flea, Ctenocephalides felis (Bouché), strains infesting cats.

To evaluate the effect of fipronil spray on adult flea mortality and flea egg production of three different cat flea, Ctenocephalides felis (Bouché) strains, 30 domestic short hair cats were randomly allocated into six groups of five cats each. On day 0, cats in groups 2, 4 and 6 were treated with fipronil at 5-6ml/kg. Cats in groups 1, 3 and 5 served as untreated controls. On days -2, 7, 14, 21, and 28 each cat was infested with 50 adult cat fleas. Groups 1 and 2 were infested with fleas from the Kansas1 Colony (KS1) strain. Groups 3 and 4 were infested with a recently colonized cat flea strain from Florida (R6). Groups 5 and 6 were infested with fleas from the ARC strain. The adulticidal activity of fipronil was determined by flea comb counts 48h after treatment and then 48h after each reinfestation. Any flea eggs produced during the infestations were collected and counted prior to the 48h comb counts. Fipronil spray was > or = 99.5% effective against adults of all three cat flea strains when applied during an active infestation. Fipronil spray provided > or = 98.2 and > or = 99.5% control of adult fleas and egg production, respectively, for all strains through week 2. On days 23 and 30 control of R6 adults and egg production was significantly lower than either the ARC or the KS1 strain. On day 30, control of R6 adults and egg production was 77.3 and 87.3%, respectively. Control of KS1 adults and egg production on day 30 was significantly lower than the ARC strain. Fipronil provided > or = 99.5 and > or = 99.9% control of ARC fleas and egg production, respectively, throughout the entire study. The susceptibility to fipronil for the three strains was also evaluated on filter paper pesticide bioassays. The R6 strain was found to be less susceptible than the KS1 and ARC strains. The LC(95) estimates for the strains were 10.13, 4.77 and 2.62mg/m(2) for the R6, ARC and KS1 strains, respectively.

Utilization of propionic acid by the L4 and adult stages of Cooperia punctata (Nematoda: Trichostrongylidae) grown in vitro.

Cultures of Cooperia punctata, a nematode parasitic in cattle, were studied in Ae medium at a pH of 7.2 to 7.3 under air. In fourth and fifth stages of development, they absorbed or otherwise took into their free pool and tissues, carbon derived from 14C-labeled sodium propionate and converted, by pathways not elucidated, a portion of this carbon into protein and lipid fractions. Thirty minutes postincubation in balanced slat solution was adequate to reduce the amount of isotope in the nematode gut to a constant level. Activity from specifically-labeled 14C-propionate was recovered (as glucose pentaacetate) from worm glucose and from CO2 evolved from cultures consisting of L4 and adult stages. Use of propionate by these worms, for whatever metabolic purpose, would result in depriving the ruminant host of some of its necessary glucogenic precursors, and could account for a specific pathogenic mechanism attendant to heavy infections with this parasite.

New host and locality record for Trypanosoma peromysci.

Trypanosoma peromysci Watson, 1912 (Sarcomastigophora: Kinetoplastida), is described from a new host and locality. One of 20 (5.0%) Peromyscus leucopus collected from Pottawatomie and Riley counties in Kansas was found to harbor the parasite. Morphometric and statistical analysis confirmed the trypanosome to be indistinguishable from T. peromysci, the only difference being a greater mean flagellar length than reported previously. This is the first reported occurrence of T. peromysci in the white-footed mouse (Peromyscus leucopus noveboracensis Fischer, 1829) and also the first record of its occurrence in Kansas.

Immunodominant proteins of Sarcocystis cruzi bradyzoites isolated from cattle affected or nonaffected with eosinophilic myositis.

Sarcocystis cruzi sarcocysts were isolated from eosinophilic myositis (EM)-affected and nonaffected bovine hearts. Isolates were ruptured and used to prepare a bradyzoite antigen extract from each heart. The nonaffected heart from one newborn calf contained no apparent sarcocysts when examined histologically and was used to prepare Sarcocystis-negative control antigen. Blood samples were taken from the heart approximately 20 minutes after slaughter. Serum was obtained and evaluated, using a radioimmunoassay to measure Sarcocystis-specific IgG and IgE titers. Sarcocystis cruzi extract from a heart without EM lesions was used for antigen in the radioimmunoassay. Sarcocystis-specific IgG titer ranged between 1:1,280 and 1:2,560 in EM-affected cattle and was 1:640 in non-affected cattle. Sarcocystis-specific IgE titer ranged between 1:640 and 1:1,280 in EM-affected and nonaffected cattle. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein (western) immunoblot analysis were used to compare antigen extracts and serum samples from EM-affected vs nonaffected cattle. Twenty protein bands, ranging from approximately 22 to 215 kD, were detected consistently on bradyzoite blots probed with anti-bovine IgG after incubation with serum samples. Seven of these bands, 37, 44, 53, 57, 94, 113, and 215 kD, were also detected consistently on bradyzoite blots probed with monoclonal anti-bovine IgE. One additional band, 61 kD, was detected consistently on bradyzoite blots probed for IgE, but was seldom recognized when probed for IgG. Sixteen protein bands were evident in silver-stained gels of S cruzi-negative, newborn calf antigen, but none were recognized by antisera on western blots.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of activity for various anthelmintics against in vitro-produced Cooperia punctata.

The in vitro-grown parasitic stages of Cooperia punctata were used to evaluate 28 compounds with different kinds and degrees of in vivo activity. Using presumptive and confirmatory tests, it was possible to establish a group order of in vitro potency that compared favorably with an order based on established in vivo use of these compounds. The procedure lends itself to evaluating activity against a given parasitic growth stage and gives a quantitative estimate (range) of the concentration that produces 50% nematode kill. The system was most successful in detecting compounds with in vivo activity for C punctata, followed in order by compounds active against Cooperia spp, other gastrointestinal nematodes of ruminants, and other nematodes of non-bovine hosts. The procedure showed some differentiation between activity against nematodes versus that against cestodes, trematodes, and arthropods. The system permits considerable flexibility in experimental design, thus making possible the acquisition of the particular information desired. In addition to establishing lethal effects on the nematode, the procedure detected compounds with nematode-anesthetizing effects. The results indicate this in vitro system can be used with some expediency as a preliminary screening method in the search for new anthelmintic compounds.

Economic value and course of infection after treatment of cattle having a low level of nematode parasitism.

To determine whether it is economically advantageous to treat calves having inapparent parasitism, we conducted experiments from 1971 to 1976, involving more than 1,800 calves from 30 pens or lots, using formulations of thiabendazole, levamisole, and crufomate (ruelene). Differential egg counts, cultured larvae, and cultured parasitic stages were used to estimate the kind and degree of nematode parasitism. Differentiation of infective larvae consistently established Cooperia as the predominating (%) genus in all fecal samplings. Bunostomum, when initially present, decreased or disappeared, whereas Trichostrongylus increased; other genera fluctuated less consistently. These qualitative generic fluctuations were not primarily the result of treatment, but more likely were seasonal variation. Judged by average daily gain (ADG), anthelmintic treatment was statistically advantageous at one or more points during the observation periods in 10 of 13 treated groups. In seven treated groups, the observation periods were concluded with statistical advantage in ADG, whereas in three groups, compensatory gain by corresponding controls had neutralized earlier advantages. The comparative influence of the various anthelmintics was not consistent from year to year. When total cost/kilogram gain was calculated from feed efficiency measuremnts and other costs, economic treatment advantage was evident in seven of 11 tests (7 of 10 treatment groups) from 1973 through 1976. This financial advantage, due primarily to feed efficiency and noted after 28 to 51 days, justified anthelmintic treatment. This advantage was not likely lost by the animals in subsequent periods (to 218 days) on pasture or in lots, since ADG indicated the treated calves performed either as well as, or better than, the nontreated controls. Considering all aspects of the study, the results indicate calves coming into Kansas from southern states and weighing 184 to 267 kg may possess a level of subclinical (symptomless) nematode parasitism that when treated will result in a response justifying expense for using anthelmintics.

Type-I hypersensitivity as a component of eosinophilic myositis (muscular sarcocystosis) in cattle.

Eight bovine hearts with lesions of eosinophilic myositis (EM) and 2 bovine hearts without EM lesions were collected at slaughter. Blood samples from these 10 hearts, and the heart of a newborn calf also were collected. Histologically, Sarcocystis cruzi was identified in the 8 hearts with EM lesions and the 2 hearts without EM lesions, but not in the heart of the newborn calf. Serum was harvested from the 10 blood samples and was used in homologous, modified, passive cutaneous anaphylaxis tests. Antigen was prepared from S cruzi bradyzoites isolated from the 2 hearts without EM lesions. Serum samples from the 8 cattle with EM lesions reacted positively to S cruzi antigen. When heat-inactivated IgE in serum (56 C for 4 hours) was used, all passive cutaneous anaphylaxis responses were considered negative. Using ELISA, serum IgE concentrations from the 10 cattle with and without EM lesions were 2.2 to 9 U/ml. As determined by radial immunodiffusion, IgM concentrations were 80 to 215 mg/dl. Immunoglobulin G concentrations were 420 to 2,050 mg/dl, but most were less than or equal to 1,700 mg/dl. Immunoglobulin A concentrations were 0 to 62 mg/dl; 1 steer with EM lesions had 0 mg/dl. Double-gel immunodiffusion confirmed the presence of Sarcocystis-specific precipitating antibodies. Sera from the 10 cattle with and without EM lesions formed at least 1 precipitin band.

The efficacy of pyrantel pamoate against ascarids and hookworms in cats.

The purposes of this study were to evaluate pyrantel pamoate administered orally at 20 mg/kg body weight for the removal of induced or natural infections of Ancylostoma tubaeformae and Toxocara cati in cats and to compare the efficacy of paste (40 mg base/g) and granule (80 mg base/g) formulations. Thirty cats of mixed breeding and various ages with natural and/or induced infections of A. tubaeformae and T. cati were assigned to one of three treatment groups: (1) non-medicated controls; (2) paste formulation at 20 mg base/kg; or (3) granule formulation at 20 mg base/kg. Infections were induced by feeding the cats on carcasses of infected mice. The study was conducted in replicates of at least one animal per treatment per replicate. The study parameters included clinical observations, physical examinations, faecal egg counts and the numbers, species and stages of worms recovered at necropsy. The paste formulation was 99.3% and 99.7% effective in reducing egg counts of Ancylostoma sp. and Toxocara sp. respectively. The granule formulation was 97.7% and 99.9% effective in reducing faecal egg counts of Ancylostoma sp. and Toxocara sp. respectively. When administered in paste form, pyrantel pamoate was 99.5% effective in removing adult Ancylostoma and 100.0% effective against adult Toxocara. The granule formulation was 97.9% effective against Ancylostoma and 100% effective against Toxocara. No toxic effects of either formulation of the drug were noted.

Demodicosis in an American bison.

An 18-month-old, male American bison (Bison bison) was presented with 7- to 9-mm size nodules periorbital, perineal, and on the ventral surface of the tail. Demodex spp. were identified from the exudate by microscopic examination. Examination 6 mo later revealed that the infestation had nearly cleared without treatment.