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Biomedical subjects

B L Blagburn

Publications and source records attributed to B L Blagburn.

At least 19 recordsLinked to original sources

World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of parasiticides for the treatment, prevention and control of flea and tick infestation on dogs and cats.

These guidelines are intended to assist the planning and conduct of laboratory and clinical studies to assess the efficacy of ectoparasiticides applied to dogs or cats for the purpose of treating, preventing and controlling flea and tick infestations. The term ectoparasiticide includes insecticidal and acaricidal compounds, as well as insect growth regulators. The range of biological activities accruing from animal treatment that are considered include: repellency and anti-feeding effects, knockdown, speed of kill, immediate and persistent lethal effects, and interference with egg fertility and subsequent development of off-host life cycle stages. Information is provided on the selection of animals, dose determination, dose confirmation and field studies, record keeping, interpretation of results and animal welfare. These guidelines are also intended to assist registration authorities involved in the approval and registration of new parasiticides, and to facilitate the worldwide adoption of harmonized procedures.

Animals↗

Determining a diagnostic dose for imidacloprid susceptibility testing of field-collected isolates of cat fleas (Siphonaptera: Pulicidae).

The susceptibility of four laboratory strains of cat fleas, Ctenocephalides felis (Bouche), to imidacloprid was determined by three different laboratories, by using a standardized bioassay protocol. The probit lines generated by the different laboratories were very similar, with LC50 values ranging from 0.32 to 0.81 ppm. Based on these data, a diagnostic dose (DD) of 3 ppm imidacloprid in larval rearing media was provisionally identified for detecting shifts in tolerance, possibly as a consequence of incipient imidacloprid resistance. None of the larvae from the susceptible laboratory strains survived the DD. Eighteen field-collected isolates were evaluated for their susceptibility to imidacloprid and to validate a DD of 3 ppm. Probit lines from 18 field-collected isolates were very similar, with LC50 values ranging from 0.14 to 1.52 ppm. When exposed to the DD, between 3 and 10% of the exposed larvae emerged as adults from only three of the 18 isolates. All other field isolates gave 100% mortality at the DD. Under the criteria established (>5% survivorship at 3 ppm), two isolates would be established on mammalian hosts and more extensive tests conducted to exclude or confirm the presence of resistance. The DD of 3 ppm is robust enough to eliminate most of the susceptible isolates collected until today, yet low enough to identify possible isolates for further testing.

Animals↗

Ponazuril inhibits the development of Eimeria vermiformis in experimentally infected outbred Swiss mice.

We evaluated a 15% paste formulation of ponazuril in outbred Swiss mice that were experimentally infected with Eimeria vermiformis. Thirty, 8-week-old female mice (approximately 20 g) were placed in one group of 10 mice and one group of 20 mice. Mice in both groups were gavaged with approximately 5,000 sporulated oocysts of E. vermiformis on day 0. Mice in group 2 (n=10) were treated orally on days 3 and 4 with ponazuril (suspended in 30% propylene glycol) at the rate of 20 mg/kg. Mice in group 1 (n=20) were gavaged with a similar volume of 30% propylene glycol. Rates of oocyst passage (oocysts/g feces) were determined on day 10 (peak patency) for treated and nontreated mice using a fecal aliquot oocyst counting technique. Oocysts were not observed in the feces of treated mice using the aliquot technique. Control mice passaged oocysts at a geometric mean rate of >104,000 oocysts/g feces. Control mice also produced significantly less feces on day 10. These results indicate that ponazuril is effective against E. vermiformis under the conditions utilized in this study, and that the E. vermiformis mouse model could be useful in predicting the efficacy of new anticoccidial drugs.

Animals↗

Molecular phylogeny of isolates of Ctenocephalides felis and related species based on analysis of ITS1, ITS2 and mitochondrial 16S rDNA sequences and random binding primers.

The phylogenetic relationships among 31 different flea isolates representing seven different species were studied by nucleotide sequence comparison of the internal transcribed spacer 1 (ITS1), internal transcribed spacer 2 (ITS2) and/or mitochondrial 16S ribosomal RNA gene (mt16S-rDNA) to examine the patterns of variation. Results show that all regions are useful in discriminating among flea species. In Ctenocephalides felis and Tunga penetrans, some differences in these gene regions occurred among different isolates within the same species. In the latter case, the differences are in the mt16S-rDNA region, with one isolate showing 48% divergence in nucleotide sequence. The taxonomic implications of this result are unclear at present. The gene regions revealed differences between C. felis isolates only after DNA sequencing the PCR products. Further differentiation among C. felis isolates was obtained using four different random binding primers (decamers) and primers for mammalian aldolase to amplify narrow differences in the genome. Using these primers we were able to discriminate between different C. felis isolates and determine that some of the genetic variation coincided with minor differences in response to the control agent imidacloprid. However, overall findings do not support the existence of subspecies of C. felis.

Animals↗

Disseminated toxoplasmosis in a captive ring-tailed lemur (Lemur catta).

A 3-yr-old secundiparous female ring-tailed lemur presented to the Auburn University Small Animal Clinic with signs of dyspnea, lethargy, and anorexia. The animal died before she could be examined, and a full necropsy was immediately performed. Provisional necropsy findings included moderate pneumonia and hepatopathy. Acute interstitial pneumonia and focal hepatocellular necrosis were confirmed histologically. Lung impression smears, histopathology, electron microscopy, immunohistochemistry, and tissue culture isolation resulted in a diagnosis of acute disseminated Toxoplasma gondii infection, which was confirmed by polymerase chain reaction. The isolate of T. gondii was avirulent for mice and was named AU Tgl and genetically is type II. The source of the infection remains unclear, but speculation suggests contaminated fruit or blackbirds (Passeriformes: Icteridae) acting as transport hosts for oocysts from nondomestic felids and feral cats on the property.

Alabama↗

Survival of nonsporulated Toxoplasma gondii oocysts under refrigerator conditions.

Toxoplasma gondii oocysts are excreted nonsporulated in the feces of the cats into the environment. These oocysts must undergo sporulation to become infectious. Little is known about the factors that influence sporulation of T. gondii oocysts. The present study examined the survival of nonsporulated oocysts under refrigerated conditions over 11-week observation period. Microscopic examination of oocysts indicated that no visible development occurred under refrigerator conditions. The nonsporulated oocysts retained their ability to sporulate when placed at room temperature. The numbers of visually viable appearing oocysts decreased over time. Some oocysts in all samples were infectious for mice despite being refrigerated for up to an 11 weeks before undergoing sporulation. Results indicate that nonsporulated oocysts can survive in the environment for at least 3 months and retain their ability to become infectious when placed under appropriate conditions.

Animals↗

Redescription of Neospora caninum and its differentiation from related coccidia.

Neospora caninum is a protozoan parasite of animals, which before 1984 was misidentified as Toxoplasma gondii. Infection by this parasite is a major cause of abortion in cattle and causes paralysis in dogs. Since the original description of N. caninum in 1988, considerable progress has been made in the understanding of its life cycle, biology, genetics and diagnosis. In this article, the authors redescribe the parasite, distinguish it from related coccidia, and provide accession numbers to its type specimens deposited in museums.

Animals↗

Development of a larval bioassay for susceptibility of cat fleas (Siphonaptera: Pulicidae) to imidacloprid.

Strategies for controlling cat fleas, Ctenocephalidesfelisfelis (Bouché), have undergone dramatic changes in the past 5 yr. With the advent of on-animal treatments with residual activity the potential for the development of insecticide resistance increases. A larval bioassay was developed to determine the baseline susceptibility of field-collected strains of cat fleas to imidacloprid. All four laboratory strains tested showed a similar level of susceptibility to imidacloprid. Advantages of this bioassay are that smaller numbers of fleas are required because flea eggs are collected for the test. Insect growth regulators and other novel insecticides can also be evaluated. Using a discriminating dose, the detection of reduced susceptibility in field strains can be determined with as few as 40 eggs.

Acetone↗

Prevalence of agglutinating antibodies to Sarcocystis neurona in raccoons, Procyon lotor, from the United States.

Equine protozoal myeloencephalitis (EPM) is the most important protozoal disease of horses in North America and it is caused by Sarcocystis neurona. Natural cases of encephalitis due to S. neurona have been reported in raccoons, Procyon lotor. We examined 99 raccoons for agglutinating antibodies to S. neurona using the S. neurona agglutination test (SAT) employing formalin-fixed merozoites as antigen. Raccoons originated in Florida (N=24, collected in 1996), New Jersey (N=25, collected in 1993), Pennsylvania (N=25, collected in 1999), and Massachusetts (N=25, collected in 1993 and 1994). We found that 58 (58.6%) of the 99 raccoons were positive for antibodies to S. neurona using the SAT; 44 of 99 raccoons (44%) had titers of > or =1:500. This prevalence is similar to the reported seroprevalence of 33-60% for S. neurona antibodies in horses from the United States using the Western blot test.

Agglutination Tests↗

Treatment of dogs infected with Hepatozoon americanum: 53 cases (1989-1998).

OBJECTIVE: To determine clinical and pathologic findings before and after short-term (group 1) and long-term (group 2) treatment in dogs with Hepatozoon americanum infection. DESIGN: Retrospective study. ANIMALS: 53 dogs with H. americanum infection. PROCEDURE: Medical records of dogs that were treated for hepatozoonosis diagnosed on the basis of meront or merozoite stages in skeletal muscle were reviewed. RESULTS: Circulating gametocytes of H. americanum were identified in 12 of 53 dogs. Dogs were treated with various drugs, including toltrazuril, trimethoprim-sulfadiazine, clindamycin, pyrimethamine, and decoquinate. Mean WBC counts prior to treatment were 85,700 and 75,200 cells/microl in groups 1 and 2, respectively, and 1 month after initiation of treatment were 12,600 and 14,600 cells/microl, respectively. Initial response to treatment was excellent in all dogs. Twenty-three of 26 dogs in group 1 relapsed at least once and died within 2 years; mean (+/- SD) survival time was 12.6+/-2.2 months. Twenty-two of 27 group-2 dogs survived; 11 dogs had no clinical signs and were still receiving decoquinate (mean duration of treatment, 21 months), 11 dogs had no clinical signs after treatment for 14 months (range, 3 to 33 months; mean survival time, 39 months [range, 26 to 53 months]), 2 dogs were lost to follow-up, and 3 dogs were euthanatized because of severe disease. CONCLUSIONS AND CLINICAL RELEVANCE: Although no treatment effectively eliminated the tissue stages of H. americanum, treatment with trimethoprim-sulfadiazine, clindamycin, and pyrimethamine followed by long-term administration of decoquinate resulted in extended survival times and excellent quality of life.

Alkaline Phosphatase↗

Molecular and phylogenetic characterisation of Cryptosporidium from birds.

Avian isolates of Cryptosporidium species from different geographic locations were sequenced at two loci, the 18S rRNA gene and the heat shock gene (HSP-70). Phylogenetic analysis of the sequence data provided support for the existence of a new avian species of Cryptosporidium infecting finches and a second species infecting a black duck. The identity of Cryptosporidium baileyi and Cryptosporidium meleagridis as valid species was confirmed. Also, C. baileyi was identified in a number of isolates from the brown quail extending the host range of this species.

Animals↗

Prevalence of agglutinating antibodies to Neospora caninum in raccoons, Procyon lotor.

Neospora caninum is an apicomplexan parasite that causes neonatal neuromuscular disease in dogs and abortions in cattle. Dogs are the only proven definitive host. Little is known about the prevalence of antibodies to this parasite in wildlife. Sera from 99 raccoons (Procyon lotor) were examined for agglutinating antibodies to N. caninum using the modified agglutination test employing formalin-fixed tachyzoites as antigen. Raccoons originated in Florida (n = 24, collected in 1996), New Jersey (n = 25, collected in 1993), Pennsylvania (n = 25, collected in 1999), and Massachusetts (n = 25, collected in 1993 and 1994). Ten (10%) had antibodies to N. caninum; 9 had titers of 1:50, and 1 (1%) had a titer of 1:100. The present study indicates that raccoons have minimal exposure to N. caninum. The sera were also tested for agglutinating antibodies to Toxoplasma gondii and 46 (46%) were positive; 16 had titers of 1:50, 8 had titers of 1:100, and 22 had titers of > or = 1:500.

Agglutination Tests↗

Neospora hughesi: experimental infections in mice, gerbils, and dogs.

Neospora hughesi is a recently described cause of equine protozoal myeloencephalitis (EPM). A rodent model for pathogenicity would facilitate development of therapies to be used in horses. In the present study, we examined the susceptibility of BALB/c gamma-interferon gene knockout (gamma-INFKO), BALB/c, CD-1, and C57BL/6 strains of mice and gerbils to infection with tachyzoites of the Nh-A1 strain of N. hughesi isolated from a horse from AL, USA. Only the gamma-IFNKO mice developed severe clinical disease following infection with N. hughesi and died 19-25 days after infection and exhibited severe cardiac lesions. In contrast, experimental infection of gamma-INFKO mice with tachyzoites of the NC-1 or NC-Liverpool strains of Neospora caninum resulted in deaths 8-10 days after infection. The most severe lesions were in the livers, spleens, and lungs of these mice. Gerbils inoculated with N. hughesi did not develop clinical disease, had few microscopic lesions, but did seroconvert. Two dogs fed the brains of mice, shown to contain N. hughesi tissue stages by cell culture and gamma-IFNKO mouse bioassay, did not shed N. caninum-like oocysts over a 23 days observation period. The marked difference in pathogenicity between the two species of Neospora in gamma-IFNKO mice, and lack of oocyst excretion by dogs fed N. hughesi infected mice provide additional evidence that the species distinction between N. caninum and N. hughesi is valid.

Agglutination Tests↗

Inoculation of Sarcocystis neurona merozoites into the central nervous system of horses.

Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome in horses from the Americas and is usually caused by infection with the apicomplexan parasite, Sarcocystis neurona. A horse model of EPM is needed to test the efficacy of chemotherapeutic agents and potential vaccines. Five horses that were negative for antibodies to S. neurona in their serum and cerebrospinal fluid (CSF) were injected in the subarachnoid space with living merozoites of the SN2 isolate of S. neurona. None of the horses developed clinical disease or died over a 132-day observation period. All five horses developed antibodies to S. neurona in their CSF and serum 3-4 weeks after injection. Two of the horses were examined at necropsy and no parasite induced lesions were observed in their tissues and no parasites were recovered from portions of their spinal cords inoculated on to cell cultures. Results of this study demonstrate that merozoites of the SN2 isolate of S. neurona will induce seroconversion but not clinical disease when inoculated directly into the CSF of nonimmune horses.

Animals↗

Evaluation of the effects of selamectin against adult and immature stages of fleas (Ctenocephalides felis felis) on dogs and cats.

The adulticidal, ovicidal, and larvicidal effects of selamectin against flea (Ctenocephalides felis felis) infestations on dogs and cats were evaluated in a series of seven controlled and masked studies (three in cats, four in dogs). Animals were randomly allocated to treatment with either selamectin at a minimum dosage of 6mgkg(-1) in the commercial formulation or one of two negative-controls (0.9% NaCl solution or the vehicle from the commercial formulation). Treatments were administered topically in a single spot on the skin at the base of the neck in front of the scapulae. Speed of kill, measured by flea comb counts at 12h intervals during the 48h immediately following a single treatment on day 0, was evaluated in two studies. One study was in dogs and the other in cats, and each animal was infested with approximately 100 unfed viable adult fleas prior to treatment. Reductions in geometric mean flea counts for selamectin compared with saline were >98% between 24 and 36h after treatment in dogs, and between 12 and 24h after treatment in cats (P< or =0.0006). Efficacy in reducing flea egg hatch and larval development was evaluated in four studies, in which dogs and cats were treated once on day 0 and then repeatedly infested with approximately 600 fleas. Flea eggs were collected approximately for 72h after each infestation, on days 3, 7, 14, 21, and 30, counted, and cultured to determine their hatchability and subsequent larval development. Compared with the vehicle, selamectin was highly effective in reducing flea egg hatch (>92% in cats) and larval development (> or =95% for dogs and cats), and emergence of adults (97.8-100% for dogs, 85.6-100% for cats) for 30 days. Effects of exposure to hair coat debris were investigated in a study with dogs treated once on day 0 and repeatedly infested with 100 adult fleas. Debris (dander, flea faeces, hair, scales) was collected on days 1, 7, 14, 21, and 30 and added to normal flea eggs or larvae for incubation. Compared with debris from vehicle-treated dogs, debris from selamectin-treated dogs was highly effective in preventing egg hatch (>96%), in killing larvae (>98%) and in preventing larval development to adults (>99%) (P</=0.0033). Selamectin was shown to be highly effective in the treatment and control of flea infestations (C. felis) on dogs and cats. The adulticidal, ovicidal, and larvicidal effects of selamectin will be important in interrupting the flea life cycle by preventing the introduction and establishment of new flea infestations in a household environment.

Administration, Topical↗

Activity of decoquinate against Cryptosporidium parvum in cell cultures and neonatal mice.

Cryptosporidium parvum is an apicomplexan parasite that is an important cause of diarrhea in neonatal calves and humans. No treatment is currently available for neonatal calves. We have recently learned from colleagues in the pharmaceutical industry that dairy practitioners are sometimes using decoquinate for the treatment of neonatal bovine cryptosporidiosis. Therefore, the present study was undertaken to determine whether the clinical observations in calves can be substantiated by laboratory investigation. Oocysts of the KSU-1 isolate of C. parvum were used to infect human ileocecal epithelial cells in vitro to measure the efficacy of treatment using an ELISA based assay. No activity was observed at 10 or 50microM decoquinate, but at 100microM an 8% inhibition of development was seen. Oocysts of the AUCp-1 isolate of C. parvum were then used to infect suckling mice. The numbers of oocysts observed in suckling mice treated with 2.5 or 5.0mg/kg decoquinate were not significantly different from untreated control suckling mice (p0.05). The results of our study suggest that decoquinate should have little efficacy for treatment of neonatal bovine cryptosporidiosis if administered once per day and that any clinical improvement observed in treated calves may be due to factors unrelated to decoquinate's effect on C. parvum.

Animals↗