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Biomedical subjects

D E Granstrom

Publications and source records attributed to D E Granstrom.

At least 37 records · Page 2Linked to original sources

Critical test evaluation (1977-1992) of drug efficacy against endoparasites featuring benzimidazole-resistant small strongyles (population S) in Shetland ponies.

Several compounds (n = 13 single or combinations; most at therapeutic dosages) were evaluated between 1977 and 1992 in critical tests (n = 91) against benzimidazole (BZ) resistant small strongyles (Population S) and several other species of internal parasites in Shetland ponies, mostly under 1 year old. The closed breeding herd, from which the test ponies were selected, had been treated every 8 weeks with cambendazole (CBZ) for 4 years (1974-1978) and oxibendazole (OBZ) for 14 years (1978-1992). Published field test data (1974-1992) on older ponies in the herd showed BZ resistance of small strongyles. Average efficacies in the present critical tests against small strongyles for OBZ (n = 59 animals) were high in early years (95% or higher), but gradually declined to a low of 1% in 1991. Side-resistance of small strongyles was evident in critical tests (n = 1-6/single drug or combination) for several other BZs and a pro-BZ; ivermectin and piperazine were highly active, but pyrantel pamoate exhibited weak activity. BZ resistance was evident for six small strongyle species (Cyathostomum catinatum, Cyathostomum coronatum, Cylicocylus nassatus, Cylicostephanus calicatus, Cylicostephanus goldi, and Cylicostephanus longibursatus). Activity on bots, ascarids, large strongyles, and pinworms was essentially as expected, indicating no drug resistance.

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A study (1977-1992) of population dynamics of endoparasites featuring benzimidazole-resistant small strongyles (population S) in Shetland ponies.

Critical tests (91) were done between 1977 and 1992 in Shetland ponies to evaluate drug susceptibility and population dynamics (present paper) of endoparasites. The test ponies, most less than 1 year old, were from a herd where older animals were treated every 8 weeks initially with cambendazole (CBZ) (1974-1978) and then with oxibendazole (OBZ) (1978-1992). Previous field test data (1974-1992) on older ponies in the breeding herd indicated the presence of benzimidazole (BZ) resistant small strongyles. Data on population dynamics from the present critical tests indicated that 28 species of small strongyles persisted over the study period in spite of initial susceptibility and later refractiveness of six species to both CBZ and OBZ. Changes in intensities and other aspects were observed for the six BZ-resistant species (Cyathostomum catinatium, Cyathostomum coronatum, Cylicocyclus nassatus, Cylicostephanus calicatus, Cylicostephanus goldi, and Cylicostephanus longibursatus). Variabilities, some striking, were found in prevalence and intensity in bots, stomach worms, ascarids, eyeworms, large strongyles, pinworms and tapeworms.

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Activity of praziquantel (0.5 mg kg-1) against Anoplocephala perfoliata (Cestoda) in equids.

Praziquantel injectable formulation was administered at 0.5 mg k-1 per os to 24 equids naturally infected with 1-183 (average 40) Anoplocephala perfoliata. Drug activity was evaluated by a modified critical test method with necropsy 24 h after treatment. There was variable efficacy of 0-100% (aggregate average 85%); for 18 equids, 93-100%, for three equids, 70-85%, and for three equids, 0-20%.

Administration, Oral↗

Recent advances in the laboratory diagnosis of equine parasitic diseases.

This article reviews recent advances in laboratory diagnosis of equine parasitic diseases. Laboratory diagnosis of most equine parasitic diseases continues to rely on standard methods. Only laboratory diagnostic tests for EPM, cryptosporidiosis, and giardiasis were included. The criteria for testing and interpretation of results for each new diagnostic method were explained. Western blot and PCR testing for EPM and immunofluorescent staining with monoclonal antibodies for cryptosporidiosis and giardiasis were reviewed.

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Identification of opossums (Didelphis virginiana) as the putative definitive host of Sarcocystis neurona.

Sarcocystis neurona is an apicomplexan that causes equine protozoal myeloencephalitis (EPM) in North and South America. Horses appear to be an aberrant host, because the merozoites continually divide in the central nervous system, without encysting. The natural host species has not previously been identified. The small subunit ribosomal RNA (SSURNA) gene of S. neurona was compared to those of Sarcocystis muris, Sarcocystis cruzi, Toxoplasma gondii, and Cryptosporidium parvum to identify a unique region suitable for a species-specific amplification primer. The S. neurona SSURNA primer was used in a polymerase chain reaction (PCR) assay for the purpose of identifying this organism in feces and intestinal digest of wildlife specimens. Sporocysts were isolated from 4 raccoons (Procyon lotor), 2 opossums (Didelphis virginiana), 7 skunks (Mephitis mephitis), 6 cats (Felis catus), 1 hawk (Accipiter sp.), and 1 coyote (Canis latrans). The S. neurona SSURNA PCR assay and a control PCR assay using protist-specific primers were applied to all sporocyst DNA samples. All sporocyst DNA samples tested positive on the control assay. The SSURNA PCR assay yielded a 484-bp product only when applied to opossum samples. The SSURNA gene of both opossum sporocyst samples was sequenced to determine its relationship to the S. neurona SSURNA gene. The sequence had 99.89% similarity with S. neurona. This suggests that opossums are the definitive host of S. neurona.

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Multifocal myositis associated with Sarcocystis sp in a horse.

Multifocal myositis was diagnosed in a 7-year-old Quarter Horse gelding on the basis of history and findings on physical examination, serum biochemical analysis, electromyography, and microscopic examination of frozen sections of muscle biopsy specimens. Histologic examination of the muscle specimen revealed multifocal accumulations of histiocytes, lymphocytes, and plasma cells, with attendant myofiber degeneration and necrosis. Parasitic cysts with morphologic characteristics of Sarcocystis sp were found in regions of myocyte degeneration and necrosis, and in regions of normal muscle. Based on a tentative diagnosis of Sarcocystis sp-induced myositis, the horse was treated with trimethoprim/sulfamethoxazole and pyrimethamine for 28 days, phenylbutazone for 5 days, and paddock rest for 30 days. At the end of treatment, the horse had gained 35 kg, its appetite had returned to normal, and muscle mass was returning to normal. Sarcocystis fayeri is the only Sarcocystis sp reported in equine muscle in the United States and is rarely associated with acute myositis or muscle atrophy. The development of clinical signs in this horse could have been the result of an underlying immunosuppression or infection with a particularly pathogenic strain or large infective dose of S fayeri.

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Differentiation of Sarcocystis neurona from eight related coccidia by random amplified polymorphic DNA assay.

Four isolates of Sarcocystis neurona from horses with equine protozoal myeloencephalitis and eight species of coccidia from the genera Sarcocystis, Toxoplasma or Eimeria were differentiated using the random amplified polymorphic DNA assay. A single, common, 550-bp DNA fragment was amplified from the DNA of each S. neurona isolate using a 16-nucleotide primer. Crosshybridization analyses among S. neurona isolates showed that DNA fragments had at least partial sequence homology. The primer generated several DNA fragments, including a 550-bp DNA fragment, from S. cruzi, Eimeria falciformis, E. neischulzi, E. ahsata and E. bovis. DNA hybridization analyses indicated no sequence homology between these fragments and the 550-bp DNA fragment generated from S. neurona. The S. neurona 550-bp DNA fragment also did not hybridize with genomic blots of various other coccidia. These results suggest that the S. neurona DNA fragment may be exploited as a species-specific probe for this parasite.

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Humoral antibody and lymphocyte blastogenesis responses in BALB/c, C3H/HeJ and AKR/N mice following Ehrlichia risticii infection.

Three strains of mice, two susceptible to Ehrlichia risticii induced disease (BALB/c and C3H/HeJ) and one resistant (AKR/N), were evaluated for the development of humoral and cell mediated immune responses following infection with E risticii. The production of serum antibody was determined by indirect fluorescent antibody testing of sera from mice of each strain challenged with one of three different dose levels of E risticii. Antibody was assayed on days 7, 9, 12, 15 and 20 after inoculation. Cell mediated immune responses were evaluated by measuring the blastogenesis response of spleen cells from E risticii infected mice 28 days after inoculation. All three strains of mice at the high challenge level responded with the production of antibody by day 9 after inoculation. Overall, the antibody response occurred earlier and was of greater magnitude in the susceptible BALB/c and C3H/HeJ strains. A marked blastogenesis response occurred in splenocytes from E risticii infected mice of all three strains upon re-exposure to ehrlichial antigen. The findings of this study indicate that susceptibility to E risticii induced disease was not the result of deficient or delayed humoral immune responses and that E risticii infection induced the development of strong cell mediated immunity.

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Transmission of some species of internal parasites in horses born in 1990, 1991, and 1992 in the same pasture on a farm in central Kentucky.

Studies were conducted on transmission of natural infections of several species of internal parasites in horses born and kept on the same pasture on a farm in central Kentucky. Data for the first year (1989) of a 4 year study on this farm have been published recently. The present research represents the second (1990), third (1991), and fourth (1992) years of the investigation. The number of animals (n = 28) examined varied from eight born in 1990 to ten each born in 1991 and 1992. For each year, examination was made of one horse per month, beginning in June of the year of birth and extending through January (1990) or March (1991 and 1992) the following year. Ages of the horses at necropsy ranged from 87 to 251 days. Major parasites present and months of recovery were: bots--Gasterophilus intestinalis in the mouth September-January and in the stomach August-March; stomach worms--Trichostrongylus axei in August and November, Habronema spp. (immature) in July-November and January, and Habronema muscae in October, January, and February; ascarids--Parascaris equorum in the small intestine and lungs all months; intestinal threadworms--Strongyloides westeri in all months except February; large strongyles--Strongylus vulgaris in the large intestine in all months except July and August and in the cranial mesenteric artery in all months, and Strongylus edentatus in the large intestine in January and in the ventral abdominal wall in all months; pinworms--Oxyuris equi in June and January-March; tapeworms--Anoplocephala perfoliata in August-October and December-March; and eyeworms--Thelazia lacrymalis August-February. Yearly differences and similarities of infections in the horses are discussed. The value of this type of research is mentioned.

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Control of cambendazole-resistant small strongyles (Population S) with oxibendazole in a pony band: an 8 year field test (1984-1992).

Studies in a band of ponies harboring Population S benzimidazole-resistant small strongyles were initiated in 1974 and have continued for 18 years. Treatment (bimonthly) was with cambendazole for the first 4 years and with oxibendazole (OBZ) for the next 14 years. Data on the first 10 years have been published. The present investigation includes the last 8 years (4 October 1984-11 September 1992), which are the seventh through fourteenth years, of treatment with OBZ. Pre- and posttreatment mean counts of strongyle eggs (epg) and larvae (lpg) per gram of feces were determined biweekly during the current study to monitor the efficacy of OBZ. The average annual percent reductions of epg counts effected by OBZ treatments were 51%, 53%, 38%, 38%, 39%, 28%, 40%, and 19% for the seventh through fourteenth years, respectively. Similar levels of reductions were observed for lpg counts. Although OBZ was initially highly effective on this population of small strongyles, epg and lpg counts gradually declined, but have remained more or less constant since the fifth year of research. However, reductions of the counts were the lowest for the last year of the study.

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Phylogenetic relationship of Sarcocystis neurona to other members of the family Sarcocystidae based on small subunit ribosomal RNA gene sequence.

Sarcocystis neurona is a coccidial parasite that causes a neurologic disease of horses in North and South America. The natural host species are not known and classification is based on ultrastructural analysis. The small subunit ribosomal RNA (SSURNA) gene of S. neurona was amplified using polymerase chain reaction techniques and sequenced by Sanger sequencing reactions. The sequence was compared with partial sequences of S. muris, S. gigantea, S. tenella, S. cruzi, S. arieticanis, S. capracanis, Toxoplasma gondii, Eimeria tenella, and Cryptosporidium parvum. Alignments of available sites for all 10 species and alignments of the entire SSURNA sequence of S. neurona, S. muris, S. cruzi, T. gondii, and C. parvum were performed. Alignments were analyzed using maximum parsimony and maximum likelihood methods to determine relative phylogeny of these organisms. These analyses confirmed placement of S. neurona in the genus Sarcocystis and suggested a close relationship to S. muris, S. gigantea, and T. gondii. Molecular phylogeny suggests that Sarcocystis spp., which utilize the dog (Canis familiaris) as the definitive host, evolved from a common ancestor, whereas those species (including T. gondii) that utilize the cat (Felis domesticus) as the definitive host evolved from another common ancestor. This suggests a possible definitive host for S. neurona.

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Natural infections of Strongyloides westeri: prevalence in horse foals on several farms in central Kentucky in 1992.

During the period 28 February-1 July 1992, fecal samples were collected and examined for eggs of Strongyloides westeri once from each of 382 horse foals (364 Thoroughbreds, 16 Standardbreds, one Lippizaner, and one draft-type horse) in central Kentucky. Ages of the foals at the time of sampling ranged from 7 to 63 days (mean 22 days). The nine farms in the study were considered to have overall excellent deworming programs. None of the foals had been treated with an antiparasitic compound before the study. Eggs of S. westeri were found in 6% (22 of 382) of the foals on 78% (7 of 9) of the farms. Current prevalence of this parasite in foals is discussed relative to studies before the availability and usage of effective drugs.

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Equine protozoal myeloencephalitis: antigen analysis of cultured Sarcocystis neurona merozoites.

Antigens of cultured Sarcocystis neurona merozoites were examined using immunoblot analysis. Blotted proteins were probed with S. cruzi, S. muris, and S. neurona antisera produced in rabbits, S. fayeri (pre- and post-infection) and S. neurona (pre- and post-inoculation) sera produced in horses, immune sera from 7 histologically confirmed cases of equine protozoal myeloencephalitis (EPM), and pre-suckle serum from a newborn foal. Eight proteins, 70, 24, 23.5, 22.5, 13, 11, 10.5, and 10 Kd, were detected only by S. neurona antiserum and/or immune serum from EPM-affected horses. Equine sera were titered by the indirect immunofluorescent antibody (IFA) method using air-dried, cultured S. neurona merozoites. Anti-Sarcocystis IFA titers were found in horses with or without EPM. Serum titers did not correspond to the number of specific bands recognized on immunoblots.

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Immunohistochemical study to demonstrate Sarcocystis neurona in equine protozoal myeloencephalitis.

A 5-year (1985-1989) retrospective immunohistochemical study was conducted using an avidin-biotin complex (ABC) immunoperoxidase method to demonstrate Sarcocystis neurona in histologically suspect cases of equine protozoal myeloencephalitis (EPM). Primary antibodies against S. neurona and S. cruzi were utilized for the ABC technique. The findings were compared with those from cases in which the organisms were detected by examination of hematoxylin and eosin (HE)-stained neuronal sections. HE-stained sections detected the presence of the organisms in 20% of the suspect cases; whereas the ABC technique confirmed the presence of S. neurona in 51% and 67% of the cases by S. neurona and S. cruzi antibodies, respectively. A review of clinical case histories showed that 21/47 (45%) of the EPM horses with parasites in the tissue sections had prior treatment with antiprotozoal drugs and/or steroids. Using the test results of S. neurona and S. cruzi as a standard reference, HE test sensitivity based on examination of up to 30 neuronal sections per case was only 25%, and test specificity was 91%.

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Critical tests of thiabendazole, oxibendazole, and oxfendazole for drug resistance of population-B equine small strongyles (1989 and 1990).

Critical tests were conducted in horses (n = 11) with naturally acquired infections of benzimidazole (BZ)-resistant population-B small strongyles in 1989 and 1990. Anthelmintics administered were thiabendazole (44 mg/kg of body weight, n = 4), oxibendazole (10 mg/kg, n = 3), and oxfendazole (OFZ, 10 mg/kg; n = 4). All compounds were paste formulations administered orally except for 1 of the OFZ treatments, which was a suspension formulation given by stomach tube. Aggregate mean efficacy was calculated for all species of small strongyles, drug-resistant and non-resistant. The highest efficacy was for oxibendazole (98%) and OFZ (94%); efficacy for thiabendazole was 63%. Five genera and 16 species of small strongyles were recovered from the 11 horses, ranging from 7 to 13 species (mean, 11). Of these, 7 species were found to have resistance in variable degrees to most of the anthelmintics. These strongyles were Cyathostomum catinatum, Cyathostomum coronatum, Cylicocyclus nassatus, Cylicostephanus calicatus, Cylicostephanus goldi, Cylicostephanus longibursatus, and Cylicostephanus minutus. The large strongyle, Strongylus vulgaris, was present in all 11 test horses, and efficacy was 100% for all drugs. Seven of the BZ-treated foals (at least 1 horse from each BZ-treatment group), were infected with S edentatus; removal was 100%.

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Critical and controlled tests of activity of moxidectin (CL 301,423) against natural infections of internal parasites of equids.

The activity of moxidectin was evaluated in 1988 and 1989 against natural infections of internal parasites in 20 critical tests (n = 20 equids) and three controlled tests (n = 20 equids). Two formulations, injectable administered intramuscularly (i.m.) or intraorally (i.o.) and gel i.o., were given at dose rates of 0.2, 0.3 or 0.4 mg kg-1 body weight. For the critical tests (all three dose rates evaluated), removals of second instar Gasterophilus intestinalis were 93-100%, except (89%) for the injectable formulation (i.m.) at 0.2 mg kg-1. Removals of third instar G. intestinalis were 88-100% for the injectable formulation given i.m. or i.o. and 93-100% for the gel formulation, except (53%) for one batch (0.4 mg kg-1). Activity was 100% for third instar Gasterophilus nasalis, Parascaris equorum, Strongylus vulgaris and Strongylus edentatus. For Oxyuris equi, removals were 91-100%, except (27%) for one batch of the injectable formulation given i.o. at 0.3 mg kg-1. There was apparent activity against migrating S. vulgaris and S. edentatus at various dose rates and routes of administration for both formulations. At necropsy, there were local reactions observed at the injection site of three equids. In the controlled tests, dose rates were 0.2 or 0.4 mg kg-1. Removal of third instar G. intestinalis was highest for the injectable formulation given i.m. All formulations and dose rates were highly effective against S. vulgaris and S. edentatus, but variable and incomplete against O. equi. Removal was excellent on Habronema muscae and on migrating S. vulgaris and S. edentatus, although incomplete on S. vulgaris. Gasterophilus nasalis third instars and P. equorum were present in low numbers in some non-treated equids, but none were recovered from treated equids. Toxicosis was not evident.

Administration, Oral↗

Equine protozoal myelitis in Panamanian horses and isolation of Sarcocystis neurona.

Schizonts of Sarcocystis neurona were identified microscopically in hematoxylin-eosin-stained spinal cord sections from 2 native Panamanian horses that exhibited clinical signs of equine protozoal myelitis (EPM). Spinal cord homogenate from a third Panamanian horse with EPM was inoculated onto monolayers of cultured bovine monocytes (M617). Intracytoplasmic schizonts containing merozoites arranged in rosette forms surrounding a central residual body first were observed 13 wk postinoculation. Parasites divided by endopolygeny and lacked rhoptries. Schizonts from each horse reacted with Sarcocystis cruzi antiserum in an immunohistochemical test.

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