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Biomedical subjects

R J MacKay

Publications and source records attributed to R J MacKay.

At least 19 recordsLinked to original sources
PubMed2003

Naturally occurring Sarcocystis infection in domestic cats (Felis catus).

Equine protozoal myeloencephalitis is an important neurological disease of horses in the United States. Consequently, there is an active research effort to identify hosts associated with the primary causative agent, Sarcocystis neurona. The purpose of this study was to determine whether the domestic cat (Felis catus) is a natural host for S. neurona. Muscle sections from 50 primarily free-roaming domestic cats were examined for the presence of sarcocysts. Serum from cats in this group and another group of 50 free-roaming cats were evaluated for the presence of S. neurona antibody. Sarcocysts were found in five of 50 (10%) cats, and S. neurona antibody in five of 100 (5%) cats. Morphological, molecular (including ribosomal RNA genes), and biological characterisation of these sarcocysts showed that they were not S. neurona or S. neurona-like. Sarcocysts found in the cats were identified morphologically as Sarcocystis felis, a common parasite of wild felids. The life cycle of S. felis is not known, and prior to this study, no molecular marker for S. felis existed. Although cats were found to be infected with S. felis sarcocysts, serological data provided evidence of possible infection with S. neurona as well. Further work is needed to determine the role of the domestic cat in the life cycle of S. neurona.

Animals↗
PubMed2001

Development of Sarcocystis falcatula in its intermediate host, the Brown-headed Cowbird (Molothrus ater).

Sporocysts of Sarcocystis falcatula obtained from experimentally infected Virginia opossums (Didelphis virginiana) were inoculated orally to 60 wild-caught Brown-headed Cowbirds (Molothrus ater). Another 30 Brown-headed Cowbirds were not challenged and served as uninfected controls. Two inoculated and one control cowbird were necropsied every 2 weeks and the pectoral and thigh muscles were examined grossly for cyst development. Stained histologic sections of pectoral muscle, thigh muscle, and lung were examined by light microscopy and presence, density, and size of sarcocysts were determined. Sarcocysts were present by 6 weeks post-inoculation (PI) and were still growing at 40 weeks PI. The sarcocysts from birds 40 weeks post-infection were infective to an opossum. The morphology of the sarcocyst wall by transmission electron microscopy substantiated the identification as S. falcatula. Lung sections were examined for the presence of schizonts, but were seen only at 2 weeks PI. This evaluation was complicated by the presence of unidentified microfilariae. These birds are migratory and the continued growth and development of muscle cysts would allow them to be a source of infection at both extremes of their geographic range, regardless of which end of the migration at which they were infected.

Animals↗
PubMed2001

Immunoconversion against Sarcocystis neurona in normal and dexamethasone-treated horses challenged with S. neurona sporocysts.

Equine protozoal myeloencephalitis is a common neurologic disease of horses in the Americas usually caused by Sarcocystis neurona. To date, the disease has not been induced in horses using characterized sporocysts from Didelphis virginiana, the definitive host. S. neurona sporocysts from 15 naturally infected opossums were fed to horses seronegative for antibodies against S. neurona. Eight horses were given 5x10(5) sporocysts daily for 7 days. Horses were examined for abnormal clinical signs, and blood and cerebrospinal fluid were harvested at intervals for 90 days after the first day of challenge and analyzed both qualitatively (western blot) and quantitatively (anti-17kDa) for anti-S. neurona IgG. Four of the challenged horses were given dexamethasone (0.1mg/kg orally once daily) for the duration of the experiment. All challenged horses immunoconverted against S. neurona in blood within 32 days of challenge and in CSF within 61 days. There was a trend (P = 0.057) for horses given dexamethasone to immunoconvert earlier than horses that were not immunosuppressed. Anti-17kDa was detected in the CSF of all challenged horses by day 61. This response was statistically greater at day 32 in horses given dexamethasone. Control horses remained seronegative throughout the period in which all challenged horses converted. One control horse immunoconverted in blood at day 75 and in CSF at day 89. Signs of neurologic disease were mild to equivocal in challenged horses. Horses given dexamethasone had more severe signs of limb weakness than did horses not given dexamethasone; however, we could not determine whether these signs were due to spinal cord disease or to effects of systemic illness. At necropsy, mild-moderate multifocal gliosis and neurophagia were found histologically in the spinal cords of 7/8 challenged horses. No organisms were seen either in routinely processed sections or by immunohistochemistry. Although neurologic disease comparable to naturally occurring equine protozoal myeloencephalitis (EPM) was not produced, we had clear evidence of an immune response to challenge both systemically and in the CNS. Broad immunosuppression with dexamethasone did not increase the severity of histologic changes in the CNS of challenged horses. Future work must focus on defining the factors that govern progression of inapparent S. neurona infection to EPM.

Animals↗
PubMed2001

The nine-banded armadillo (Dasypus novemcinctus) is an intermediate host for Sarcocystis neurona.

The nine-banded armadillo (Dasypus novemcinctus) is an intermediate host of at least three species of Sarcocystis, Sarcocystis dasypi, Sarcocystis diminuta, and an unidentified species; however, life cycles of these species have not been determined. Following feeding of armadillo muscles containing sarcocysts to the Virginia opossum (Didelphis virginiana), the opossums shed sporulated Sarcocystis sporocysts in their faeces. Mean dimensions for sporocysts were 11.0x7.5 microm and each contained four sporozoites and a residual body. Sporocysts were identified as Sarcocystis neurona using PCR and DNA sequencing. A 2-month-old foal that was negative for S. neurona antibodies in the CSF was orally inoculated with 5x10(5) sporocysts. At 4 weeks post-infection, the foal had a 'low positive' result by immunoblot for CSF antibodies to S. neurona and by week 6 had a 'strong positive' CSF result and developed an abnormal gait with proprioceptive deficits and ataxia in all four limbs. Based on the results of this study, the nine-banded armadillo is an intermediate host of S. neurona.

Animals↗
PubMed2001

The nine-banded armadillo (Dasypus novemcinctus) is naturally infected with Sarcocystis neurona.

Sarcocysts were dissected from the tongue of a nine-banded armadillo (Dasypus novemcinctus). DNA was extracted and characterised by PCR amplification followed by restriction fragment length polymorphism analysis and nucleotide sequencing. A total of 1879 nucleotides were compared; the sarcocyst DNA sequence was identical to that reported for Sarcocystis neurona. DNA was extracted from the sarcocysts of five more nine-banded armadillos. A 254-nucleotide sequence was determined for each and found to be identical to S. neurona. Western blot techniques for detection of anti-S. neurona antibody were developed for use with armadillo plasma and samples from 19 wild-caught and 17 captive-raised armadillos were examined. Whereas all of the 19 wild-caught armadillos had antibodies to S. neurona, only one of 17 captive-raised armadillos did. These results suggest that the nine-banded armadillo are naturally infected with S. neurona.

Animals↗
PubMed2001

The striped skunk (Mephitis mephitis) is an intermediate host for Sarcocystis neurona.

Striped skunks, initially negative for antibodies to Sarcocystis neurona, formed sarcocysts in skeletal muscles after inoculation with S. neurona sporocysts collected from a naturally infected Virginia opossum (Didelphis virginiana). Skunks developed antibodies to S. neurona by immunoblot and muscles containing sarcocysts were fed to laboratory-reared opossums which then shed sporulated Sarcocystis sporocysts in their faeces. Mean dimensions for sporocysts were 11.0 x 7.5 microm and each contained four sporozoites and a residuum. Sarcocysts from skunks and sporocysts from opossums fed infected skunk muscle were identified as S. neurona using PCR and DNA sequence analysis. A 2-month-old, S. neurona-naive pony foal was orally inoculated with 5 x 10(5) sporocysts. Commercial immunoblot for antibodies to S. neurona performed using CSF collected from the inoculated pony was low positive at 4 weeks p.i., positive at 6 weeks p.i., and strong positive at 8 weeks p.i. Gamma-interferon gene knockout mice inoculated with skunk/opossum derived sporocysts developed serum antibodies to S. neurona and clinical neurologic disease. Merozoites of S. neurona present in the lung, cerebrum, and cerebellum of mice were detected by immunohistochemistry using polyclonal antibodies to S. neurona. Based on the results of this study, the striped skunk is an intermediate host of S. neurona.

Animals↗
PubMed2000

Equine protozoal myeloencephalitis.

Recent advances in the understanding of the parasite life cycle, epidemiology, clinical signs, diagnosis, treatment, and prevention of EPM are reviewed. The NAHMS Equine '98 study and a controlled retrospective study from The Ohio State University College of Veterinary Medicine identified a number of risk factors associated with development of the disease. The national annual incidence of EPM was 1% or less depending on the primary use of the animals. Increased disease risk was associated with age (1-5 and > 13 years of age), season (lowest in winter months and increasing with ambient temperature), previous stressful events, the presence of opossums, the use of nonsurface water drinking systems, and failure to restrict wildlife access to feed. Horses that received treatment were 10 times more likely to improve, and those that improved were 50 times more likely to survive. A number of recent studies confirmed that horses can be experimentally infected with S. neurona; however, large numbers of sporocysts are apparently necessary to achieve infection, and clinical signs and abnormal CNS histology are only seen inconsistently. Results suggest that CNS infection and positive CSF immunoblot findings may be transient phenomena among naturally infected horses. Although immunosuppression may be involved in the development of EPM, some element of the immune response seems to be necessary for the development of clinical signs. Use of the standard immunoblot test for the detection of anti-S. neurona antibodies in CSF continues to provide the most useful adjunct to a detailed neurologic examination for the diagnosis of EPM. Test sensitivity and specificity were 89% in 295 horses euthanatized because of neurologic disease, of which 123 were confirmed cases of EPM. The PPV was 85%, and the NVP was 92%. A number of promising new EPM treatments are under investigation. In addition to standard SDZ/PYR therapy, toltrazuril, ponazuril, diclazuril, and NTZ have shown promise as possible alternatives.

Animals↗
PubMed2000

Inflammation in horses.

After inflammation is initiated by detection of antigen, plasma components and activated leukocytes are concentrated at the inflammatory site. Cellular and chemical effectors of inflammation are focused on the offending antigen, usually resulting in its destruction and elimination. Activation of endogenous counter-regulatory systems damps down the inflammatory process and is the first stage of repair. In addition to local effects, the inflammatory focus may initiate a continuum of systemic acute phase responses ranging from the systemic inflammatory response syndrome (SIRS) to generalized immunosuppression.

Animals↗
PubMed2000

Effect of eltenac in horses with induced endotoxaemia.

Ten horses were used in a crossover study to evaluate the effectiveness of eltenac against endotoxaemia. Eltenac (0.5 mg/kg bwt) or saline control was given i.v. then 15 min later, intravenous infusion of endotoxin was begun and continued for 120 min (total dose 100 ng/kg bwt). Horses were monitored for heart and respiratory rates, pulmonary and carotid arterial pressure and core body temperature. Blood was sampled at intervals for measurement of haematological variables and plasma concentrations of lactate, prostanoid metabolites, tumour necrosis factor (TNF) and stress hormones. In comparison with saline-treatment, use of eltenac significantly protected against endotoxin-induced changes in respiratory rate, core temperature, systemic arterial blood pressure (SAP), pulmonary arterial pressure, PCV, and plasma protein, 6-keto prostaglandin F1 alpha, thromboxane B2, epinephrine, and cortisol concentrations. Despite statistical effect of eltenac on SAP, values in both treatment groups remained well above baseline throughout the evaluation period. Significant protective effect of eltenac was not found for heart rate, white blood cell count, plasma lactate concentration or TNF activity. On the basis of these results, it is expected that use of eltenac will provide clinical benefit in horses with naturally occurring endotoxaemia.

Aniline Compounds↗
PubMed2000

Small intestinal adenomatous polyposis resulting in protein-losing enteropathy in a horse.

A 4-year-old Quarter Horse gelding was presented with a history of weight loss of 6 months duration, along with extensive ventral subcutaneous edema. Clinicopathologic findings included a markedly low serum total protein (2.9 g/dl) and a low packed cell volume (24%). The mucosal surface of the distal jejunum and entire ileum were carpeted with numerous polypoid, papillary, and glandular masses comprised of pseudostratified tall columnar cells and large numbers of interspersed goblet cells. Neoplastic change was diffuse throughout the mucosa of each mass, but abrupt demarcation occurred between neoplastic masses and adjacent mucosa. Immunohistochemical staining for protein of the p53 tumor suppressor gene revealed only occasional cytoplasmic reactivity within polyps and normal mucosa. Nuclear staining for papillomavirus antigens was not observed. Electron microscopic examination revealed features of well-differentiated intestinal epithelial cells, including apical tight junctions and microvilli, desmosomes, and the presence of numerous goblet cells. Microorganisms were not detected. Small intestinal polyposis should be considered as a rare differential diagnosis for protein-losing enteropathy in the horse.

Adenomatous Polyps↗
PubMed1999

Multiple DNA markers differentiate Sarcocystis neurona and Sarcocystis falcatula.

Studies designed to investigate the causative agent of equine protozoal myeloencephalitis and its life cycle have been hampered by the marked similarity of Sarcocystis neurona to other Sarcocystis spp. present in the same definitive host. Random-amplified polymorphic DNA techniques were used to amplify DNA from isolates of S. neurona and Sarcocystis falcatula. DNA sequence analysis of polymerase chain reaction (PCR) products was then used to design PCR primers to amplify specific Sarcocystis spp. DNA products. The ribosomal RNA internal transcribed spacer was also amplified and compared between S. neurona and S. falcatula. Useful sequence heterogeneity between the 2 organisms was identified, creating potential markers to distinguish these Sarcocystis spp. These markers were used to characterize Sarcocystis isolates from opossum (Didelphis virginiana) feces. Our data suggest that S. neurona and S. falcatula can be differentiated with these markers and that multiple Sarcocystis spp., including S. neurona and S. falcatula, are shed by opossums.

Animals↗
PubMed1999

Are Sarcocystis neurona and Sarcocystis falcatula synonymous? A horse infection challenge.

Equine protozoal myeloencephalitis (EPM) is a debilitating neurologic disease of the horse. The causative agent. Sarcocystis neurona, has been suggested to be synonymous with Sarcocystis falcatula, implying a role for birds as intermediate hosts. To test this hypothesis, opossums (Didelphis virginiana) were fed muscles containing S. falcatula sarcocysts from naturally infected brown-headed cowbirds (Molothrus ater). Ten horses were tested extensively to ensure no previous exposure to S. neurona and were quarantined for 14 days, and then 5 of the horses were each administered 10(6) S. falcatula sporocysts collected from laboratory opossums. Over a 12-wk period, 4 challenged horses remained clinically normal and all tests for S. neurona antibody and DNA in serum and cerebrospinal fluid were negative. Rechallenge of the 4 seronegative horses had identical results. Although 1 horse developed EPM, presence of S. neurona antibody prior to challenge strongly indicated that infection occurred before sporocyst administration. Viability of sporocysts was confirmed by observing excystation in equine bile in vitro and by successful infection of naive brown-headed cowbirds. These data suggest that S. falcatula and S. neurona are not synonymous. One defining distinction is the apparent inability of S. falcatula to infect horses, in contrast to S. neurona, which was named when cultured from equine spinal cord.

Animals↗
PubMed1999

Effect of a conjugate of polymyxin B-dextran 70 in horses with experimentally induced endotoxemia.

OBJECTIVE: To determine the efficacy of polymyxin B-dextran 70 (PBD) for treatment of endotoxemic horses. ANIMALS: 15 horses during study 1 and 6 horses during study 2. PROCEDURES: 3 groups were used in study 1. Horses in groups 1 and 2 were given 30 ng of lipopolysaccharide (LPS)/kg of body weight, IV, over 60 minutes. Horses in group 3 were given saline (0.9% NaCl) solution. Beginning 15 minutes before LPS infusion and continuing for 75 minutes, horses in groups 1 and 3 were given PBD, IV. Horses in group 2 were given dextran 70. Blood samples were obtained for hemograms and determination of cytokine, lactate, and prostanoid concentrations. In study 2, horses were given ketoprofen (2.2 mg/kg) or saline solution 15 minutes before infusion of PBD. Fourteen days later, treatments were reversed, using a crossover design. Blood samples were obtained for measurement of thromboxane B2 (TXB2) concentration. RESULTS: For study 1, prior treatment with PBD completely blocked endotoxin-induced changes for heart and respiratory rates, rectal temperature, WBC count, and plasma tumor necrosis factor, interleukin 6, TXB2, and prostaglandin F1 concentrations. There was transient tachypnea, sweating, and increased plasma TXB2 concentration in horses given PBD (with or without LPS). Prior treatment with ketoprofen eliminated all PBD-induced signs and prevented the increase in plasma TXB2 concentration. CONCLUSIONS: Signs of endotoxemia were prevented in horses by treatment with PBD, although its use was associated with mild adverse effects. CLINICAL RELEVANCE: When used in combination with a cyclooxygenase-inhibiting drug, PBD has potential for treatment of horses with endotoxemia.

Animals↗
PubMed1998

Human interleukin 10 suppresses production of inflammatory mediators by LPS-stimulated equine peritoneal macrophages.

To investigate the ability of recombinant human interleukin 10 (rhuIL-10) to suppress the release of inflammatory mediators from lipopolysaccharide (LPS)-stimulated equine macrophages, rhuIL-10 was added to equine peritoneal macrophage monolayers at concentrations of 0, 0.1, 1, 10, or 100 ng/ml. Thirty minutes later, LPS (E. coli O55:B5) was added at final concentrations of 0, 1, 10, 100 ng/ml. Macrophages were incubated for 16 h at 37 degrees C, then supernates were harvested and assayed for tumor necrosis factor (TNF) activity (L929 cytotoxicity), interleukin-6 (IL-6) activity (B9 proliferation), prostaglandin E2 concentration (ELISA), and nitric oxide (Griess reaction for nitrite). Preincubation of LPS-stimulated peritoneal macrophages with rhuIL-10 caused significant (P<0.05) reduction in secretion of TNF, IL-6, and PGE2, in a dose-dependent manner. Of the inflammatory mediators, TNF was most sensitive to the effects of rhuIL-10. At concentrations of rhuIL-10> or =1 ng/ml, TNF activity in the supernate was inhibited significantly at all concentrations of LPS. At one or more LPS concentrations, there was significant inhibition of each mediator in the presence of 1 ng rhuIL-10/ml and, at 100 ng/ml, rhuIL-10 significantly inhibited production of each mediator at all LPS concentrations tested. When data were expressed as a percentage of control values and pooled across all LPS concentrations, both PGE2 and TNF values were significantly reduced at rhuIL-10 concentrations of > or =1 ng/ml, whereas IL-6 was inhibited significantly at concentrations of > or =10 ng rhuIL-10/ml. Tumor necrosis factor production was more completely suppressed (7.8% of control) by the highest concentration of rhuIL-10(100 ng/ml) than was PGE2 (27.2%) or IL-6 (43.8%). Nitrite was not detected in any supernate from peritoneal macrophage monolayers.

Animals↗
PubMed1997

Overwhelming strongyloidosis in a foal.

A 6-month-old foal was evaluated because of weakness, weight loss, and inappetence of 3 weeks' duration. On initial examination, the foal was weak, poorly responsive, and emaciated. Clinicopathologic abnormalities included anemia and hypoproteinemia. Because of its severe debilitation, the foal was euthantized. Necropsy revealed marked infection of the small intestine with Strongyloides westeri and severe edema of the entire gastrointestinal tract. The foal had been orphaned when it was 6 hours old and raised in isolation from other horses. We believe that this foal developed overwhelming strongyloidosis when it was first exposed to other foals at 5 months of age, because it had not been naturally exposed to the organism at a younger age and was immunologically naive.

Animals↗
PubMed1997

Equine protozoal myeloencephalitis.

Equine protozoal myeloencephalitis (EPM) is a common neurologic disease of horses in the Americas. Horses with EPM most commonly have abnormalities of gait, but they also may present with signs of brain disease. The disease ranges in severity from mild lameness to sudden recumbency, and clinical signs usually are progressive. A causative agent, Sarcocystis neurona, has been isolated from affected horses, and serologic surveys suggest that approximately 50% of horses in the United States have been exposed. EPM is considered a treatable disease, although the response to antimicrobial treatment often is incomplete. This article highlights new information about the life cycle of S. neurona and reviews the literature regarding diagnosis, clinical signs, and treatment of the disease.

Animals↗