RESUMO
Among the recognized neurologic diseases in horses, equine protozoal myeloencephalitis (EPM) has been reported around the world and still presents challenges in diagnosis and treatment. Horses can present with clinical neurologic signs consistent with EPM while testing negative for the two main causative agents, Sarcocystis neurona or Neospora hughesi, and may still be clinically responsive to anti-parasitic drug therapy. This context led to our hypothesis that another protozoal parasite, Toxoplasma gondii, which is known to cause toxoplasmosis in other mammalian species, is a potential pathogen to cause neurologic disease in horses. To evaluate this hypothesis, serum and cerebrospinal fluid (CSF) were collected from 210 horses presenting with clinical signs compatible with EPM, and the indirect immunofluorescent antibody test (IFAT) was used to detect antibody titers for T. gondii, S. neurona, and N. hughesi. Additionally, the serum to CSF titer ratio was calculated for T. gondii, S. neurona, and N. hughesi infections, suggesting intrathecally-derived antibodies for each of the three agents if the serum:CSF ratio was ≤ 64. There were 133 (63.3%) horses positive for serum T. gondii antibodies using a cutoff titer of 160, and 31 (14.8%) positive for CSF T. gondii antibodies using a cutoff titer of 5. Overall, 21 (10.0%) of EPM-suspect horses had a serum:CSF ratio ≤ 64 for antibodies for T. gondii, while 43 (20.5%) and 8 (3.8%) horses had a serum to CSF ratio ≤ 64 for antibodies for S. neurona and N. hughesi, respectively. A total of 6 (2.9%) animals presented evidence of concurrent intrathecally-derived antibodies for T. gondii and at least one other apicomplexan parasite in this study. Signalment and clinical signs were not different across the groups aforementioned. These data provide evidence of intrathecal production of anti-T. gondii antibodies, indicative of T. gondii infection in the brain and/or spinal cord of horses with EPM-like disease.
Assuntos
Encefalomielite , Doenças dos Cavalos , Sarcocystis , Sarcocistose , Toxoplasma , Cavalos , Animais , Sarcocistose/veterinária , Sarcocistose/parasitologia , Anticorpos Antiprotozoários , Doenças dos Cavalos/diagnóstico , Encefalomielite/veterinária , Encefalomielite/parasitologia , MamíferosRESUMO
Interactions with livestock in public settings such as county and state fairs can expose people and other livestock to faecal material capable of spreading zoonotic enteric pathogens. The goal of this study was to understand these risks by screening livestock faeces (n = 245) and livestock bedding (n = 155) for common zoonotic pathogens (Giardia, Cryptosporidium, Salmonella and Campylobacter spp.) and by measuring faecal indicator, Escherichia coli, concentrations in drinking water (n = 153), feed containers (n = 124) and bedding material (n = 157) in four livestock species (cattle, sheep, goats and swine) from county fairs in California, USA. Results indicated that sheep were most likely to have pathogens detected in faeces and that Giardia was the most frequently detected pathogen in both faeces (11%) and bedding (21%) across all livestock species. Additionally, increasing the number of animals in a holding pen at fairs, increasing the stocking density of animals in transport trailers to fairs, and having access to water in transport trailers significantly increased the odds of detecting pathogens in livestock faeces of any animal species. Observing solid material in water, stale feed and soiled bedding was associated with detecting higher E. coli concentrations. These findings provide evidence of faecal pathogens present at county fairs and suggest that site observations can aid in assessing levels of faecal exposure. The findings also indicate that the use of biosecurity measures such as (a) routine changing of livestock drinking water, feed and bedding, (b) not overstocking animals in holding pens and trailers and (c) keeping species in separate holding areas may reduce the risk of humans and livestock being exposed to faecal pathogens.
Assuntos
Doenças dos Bovinos , Criptosporidiose , Cryptosporidium , Doenças das Cabras , Doenças dos Ovinos , Doenças dos Suínos , Adolescente , Animais , Biosseguridade , Bovinos , Escherichia coli , Fezes , Humanos , Ovinos , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
An ante-mortem diagnosis of equine protozoal myeloencephalitis (EPM) is presently based on clinical presentation, immunodiagnostics performed on serum and cerebrospinal fluid (CSF), and ruling out other neurological disorders. Molecular techniques introduce a novel and promising approach for the detection of protozoal agents in CSF. Hypothesizing that real-time PCR (rtPCR) can be a useful complement to EPM diagnostics, 210 CSF samples from horses suspected of neurological disease with EPM included as a differential diagnosis were tested using rtPCR to detect Sarcocystis neurona DNA and immunodiagnostics targeting antibodies against the same pathogen, performed on serum and CSF samples. Molecular and immunological results were compared with respect to origin of the horse, time of the year, signalment, clinical signs and treatment history. Twenty-five horses tested positive in CSF for S. neurona by rtPCR only, while 30 horses had intrathecally-derived antibodies to S. neurona only (serum to CSF ratio ≤ 64 by indirect fluorescent antibody test - IFAT), and 13 horses tested rtPCR-positive in CSF with evidence of intrathecally-derived antibodies to S. neurona. Previous treatment for EPM was the only variable presenting statistical difference between the two testing modalities, highlighting that animals with history of anti-protozoal treatment were more likely to test positive solely in IFAT, while horses without treatment were more likely to test positive by rtPCR only. The results support the use of molecular diagnosis for EPM caused by S. neurona as a complement to immunodiagnostics. The use of rtPCR in CSF for the detection of S. neurona may improve the diagnostic work-up of neurologic disease suspected horses, especially in animals without previous anti-protozoal treatment.
Assuntos
Doenças dos Cavalos/líquido cefalorraquidiano , Doenças dos Cavalos/parasitologia , Doenças do Sistema Nervoso/parasitologia , Sarcocystis/genética , Sarcocistose/veterinária , Animais , DNA de Protozoário/líquido cefalorraquidiano , Cavalos , Doenças do Sistema Nervoso/patologia , Patologia Molecular , Sarcocistose/líquido cefalorraquidiano , Sarcocistose/complicações , Sarcocistose/parasitologiaRESUMO
The western blacklegged tick, Ixodes pacificus, an important vector in the western United States of two zoonotic spirochetes: Borrelia burgdorferi (also called Borreliella burgdorferi), causing Lyme disease, and Borrelia miyamotoi, causing a relapsing fever-type illness. Human cases of Lyme disease are well-documented in California, with increased risk in the north coastal areas and western slopes of the Sierra Nevada range. Despite the established presence of B. miyamotoi in the human-biting I. pacificus tick in California, clinical cases with this spirochete have not been well studied. To assess exposure to B. burgdorferi and B. miyamotoi in California, and to address the hypothesis that B. miyamotoi exposure in humans is similar in geographic range to B. burgdorferi, 1,700 blood donor sera from California were tested for antibodies to both pathogens. Sampling was from high endemic and low endemic counties for Lyme disease in California. All sera were screened using the C6 ELISA. All C6 positive and equivocal samples and nine randomly chosen C6 negative samples were further analyzed for B. burgdorferi antibody using IgG western blot and a modified two ELISA test system and for B. miyamotoi antibody using the GlpQ ELISA and B. miyamotoi whole cell sonicate western blot. Of the 1,700 samples tested in series, eight tested positive for antibodies to B. burgdorferi (0.47%, Exact 95% CI: 0.20, 0.93) and two tested positive for antibodies to B. miyamotoi (0.12%, Exact 95% CI: 0.01, 0.42). There was no statistically significant difference in seroprevalence for either pathogen between high and low Lyme disease endemic counties. Our results confirm a low frequency of Lyme disease and an even lower frequency of B. miyamotoi exposure among adult blood donors in California; however, our findings reinforce public health messaging that there is risk of infection by these emerging diseases in the state.
Assuntos
Doadores de Sangue , Borrelia burgdorferi/patogenicidade , Borrelia/patogenicidade , Doença de Lyme/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Borrelia/isolamento & purificação , Borrelia burgdorferi/isolamento & purificação , California/epidemiologia , Feminino , Humanos , Doença de Lyme/epidemiologia , Doença de Lyme/parasitologia , Doença de Lyme/transmissão , Masculino , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Adulto JovemRESUMO
OBJECTIVE To describe the general seroprevalence of anti-Sarcocystis neurona and anti-Neospora hughesi antibodies among healthy equids by use of indirect fluorescent antibody tests and determine potential risk factors for seropositivity. DESIGN Cross-sectional study. SAMPLE Whole blood samples collected from 5,250 equids (1 sample/animal) across 18 states in the United States during October 2013. PROCEDURES Information regarding potential risk factors (geographic region, breed, primary use, sex, and age) was collected along with the blood samples. For each equid, an indirect fluorescent antibody test was used to determine serum titers of antibody against each of the 2 protozoal parasites. Mixed-effects logistic regression models were created to determine ORs for seropositivity. RESULTS The overall seroprevalence of anti-S neurona and anti-N hughesi antibodies in the tested equids was 78% and 34%, respectively. Of the equids, 31% were seropositive and 18% were seronegative for antibodies against both parasites. Factors associated with equids being seropositive for anti-S neurona antibodies were residence in the South, warmblood breed, and age > 5 years. Seroprevalence of anti-N hughesi antibodies did not differ among equids in different states across the country, but warmblood breed and age > 5 years were associated with seropositivity. CONCLUSIONS AND CLINICAL RELEVANCE With regard to risk factors for S neurona and N hughesi exposure and antibody response among tested equids, older age was not unexpected; however, the influences of warmblood breed and geographic location on seropositivity for anti-S neurona antibody but not for anti-N hughesi antibody deserve further investigation.
Assuntos
Coccidiose/veterinária , Encefalomielite/veterinária , Doenças dos Cavalos/epidemiologia , Neospora/isolamento & purificação , Sarcocystis/isolamento & purificação , Sarcocistose/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Coccidiose/epidemiologia , Coccidiose/parasitologia , Estudos Transversais , Encefalomielite/epidemiologia , Encefalomielite/parasitologia , Feminino , Doenças dos Cavalos/sangue , Doenças dos Cavalos/parasitologia , Cavalos , Masculino , Neospora/imunologia , Prevalência , Fatores de Risco , Sarcocystis/imunologia , Sarcocistose/epidemiologia , Sarcocistose/parasitologia , Estados Unidos/epidemiologiaRESUMO
Tissue-cyst forming coccidia in the family Sarcocystidae are etiologic agents of protozoal encephalitis in marine mammals including the federally listed Southern sea otter (Enhydra lutris). California sea lions (Zalophus californianus), whose coastal habitat overlaps with sea otters, are definitive hosts for coccidian protozoa provisionally named Coccidia A, B and C. While Coccidia A and B have unknown clinical effects on aquatic wildlife hosts, Coccidia C is associated with severe protozoal disease in harbor seals (Phoca vitulina). In this study, we conducted surveillance for protozoal infection and fecal shedding in hospitalized and free-ranging California sea lions on the Pacific Coast and examined oocyst morphology and phenotypic characteristics of isolates via mouse bioassay and cell culture. Coccidia A and B were shed in similar frequency, particularly by yearlings. Oocysts shed by one free-ranging sea lion sampled at Año Nuevo State Park in California were previously unidentified in sea lions and were most similar to coccidia infecting Guadalupe fur seals (Arctocephalus townsendi) diagnosed with protozoal disease in Oregon (USA). Sporulated Coccidia A and B oocysts did not replicate in three strains of mice or in African green monkey kidney cells. However, cultivation experiments revealed that the inoculum of fecally-derived Coccidia A and B oocysts additionally contained organisms with genetic and antigenic similarity to Sarcocystis neurona; despite the absence of detectable free sporocysts in fecal samples by microscopic examination. In addition to the further characterization of Coccidia A and B in free-ranging and hospitalized sea lions, these results provide evidence of a new role for sea lions as putative mechanical vectors of S. neurona, or S. neurona-like species. Future work is needed to clarify the distribution, taxonomical status, and pathogenesis of these parasites in sea lions and other marine mammals that share their the near-shore marine environment.
RESUMO
Toxoplasma gondii and Sarcocystis neurona are protozoan parasites with terrestrial definitive hosts, and both pathogens can cause fatal disease in a wide range of marine animals. Close monitoring of threatened southern sea otters (Enhydra lutris nereis) in California allowed for the diagnosis of dual transplacental transmission of T. gondii and S. neurona in a wild female otter that was chronically infected with both parasites. Congenital infection resulted in late-term abortion due to disseminated toxoplasmosis. Toxoplasma gondii and S. neurona DNA was amplified from placental tissue culture, as well as from fetal lung tissue. Molecular characterization of T. gondii revealed a Type X genotype in isolates derived from placenta and fetal brain, as well as in all tested fetal organs (brain, lung, spleen, liver and thymus). This report provides the first evidence for transplacental transmission of T. gondii in a chronically infected wild sea otter, and the first molecular and immunohistochemical confirmation of concurrent transplacental transmission of T. gondii and S. neurona in any species. Repeated fetal and/or neonatal losses in the sea otter dam also suggested that T. gondii has the potential to reduce fecundity in chronically infected marine mammals through parasite recrudescence and repeated fetal infection.
Assuntos
Aborto Animal/etiologia , Lontras/parasitologia , Sarcocistose/veterinária , Toxoplasmose Animal/congênito , Toxoplasmose Animal/complicações , Animais , Anticorpos Antiprotozoários/sangue , California , Células Cultivadas , Doença Crônica , DNA de Protozoário/análise , Feminino , Genótipo , Gravidez , Sarcocystis/genética , Sarcocystis/fisiologia , Sarcocistose/complicações , Sarcocistose/congênito , Sarcocistose/transmissão , Toxoplasma/genética , Toxoplasma/fisiologia , Toxoplasmose Animal/patologia , Toxoplasmose Animal/transmissãoRESUMO
Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi being considered nonendemic on the U.S. mainland. The purpose of the present study was to evaluate the suitability of RAP-1-cELISA as a single serological test to determine the infection status of B. caballi in U.S. horses. Immunoblotting indicated that sera from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein. Antibody reactivity to B. caballi lysate was exclusively directed against a single â¼50-kDa band corresponding to a native B. caballi RAP-1 protein. In contrast, sera from experimentally and naturally infected horses from regions where B. caballi is endemic bound multiple proteins ranging from 30 to 50 kDa. Dilutions of sera from U.S. horses positive by cELISA revealed low levels of antibodies, while sera from horses experimentally infected with B. caballi and from areas where B. caballi is endemic had comparatively high antibody levels. Finally, blood transfer from seropositive U.S. horses into naive horses demonstrated no evidence of B. caballi transmission, confirming that antibody reactivity in cELISA-positive U.S. horses was not consistent with infection. Therefore, we conclude that a combination of cELISA and immunoblotting is required for the accurate serodiagnosis of B. caballi.
Assuntos
Anticorpos Antiprotozoários/sangue , Babesia/imunologia , Babesiose/veterinária , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/imunologia , Proteínas de Protozoários , Animais , Antígenos de Protozoários/imunologia , Babesiose/diagnóstico , Babesiose/imunologia , Ensaio de Imunoadsorção Enzimática , Cavalos , Proteínas de Protozoários/imunologia , Soro/química , Estados UnidosRESUMO
During April 2004, 40 sick and dead southern sea otters (Enhydra lutris nereis) were recovered over 18km of coastline near Morro Bay, California. This event represented the single largest monthly spike in mortality ever recorded during 30 years of southern sea otter stranding data collection. Because of the point-source nature of the event and clinical signs consistent with severe, acute neurological disease, exposure to a chemical or marine toxin was initially considered. However, detailed postmortem examinations revealed lesions consistent with an infectious etiology, and further investigation confirmed the protozoan parasite Sarcocystis neurona as the underlying cause. Tissues from 94% of examined otters were PCR-positive for S. neurona, based on DNA amplification and sequencing at the ITS-1 locus, and 100% of tested animals (n=14) had elevated IgM and IgG titers to S. neurona. Evidence to support the point-source character of this event include the striking spatial and temporal clustering of cases and detection of high concentrations of anti-S. neurona IgM in serum of stranded animals. Concurrent exposure to the marine biotoxin domoic acid may have enhanced susceptibility of affected otters to S. neurona and exacerbated the neurological signs exhibited by stranded animals. Other factors that may have contributed to the severity of this epizootic include a large rainstorm that preceded the event and an abundance of razor clams near local beaches, attracting numerous otters close to shore within the affected area. This is the first report of a localized epizootic in marine wildlife caused by apicomplexan protozoa.
Assuntos
Organismos Aquáticos/parasitologia , Epidemias , Lontras/parasitologia , Sarcocystis , Sarcocistose/epidemiologia , Animais , Anticorpos Antiprotozoários/sangue , Bivalves/química , Encéfalo/parasitologia , California , DNA Espaçador Ribossômico/genética , Ácido Caínico/análogos & derivados , Ácido Caínico/análise , Músculo Esquelético/química , Músculo Esquelético/parasitologia , Fármacos Neuromusculares Despolarizantes/análise , Oceano Pacífico , Sarcocystis/genética , Sarcocistose/mortalidade , Sarcocistose/patologiaRESUMO
To investigate how different routes of Toxoplasma gondii transmission influence the antibody response and infection status of deer mice (Peromyscus maniculatus), 80 mice were orally infected with 1, 5, 10, or 100 T. gondii oocysts. Ten weeks postinfection, 15 T. gondii -seropositive female mice were bred and allowed to produce 2 litters. Evidence of persistent T .gondii infection in orally infected mice was detected by serology and DNA amplification in mice from all 4 oocyst treatment groups, including those that received only a single T. gondii oocyst. Congenital transmission of T. gondii was detected by polymerase chain reaction (PCR) in 7/8 first and 4/7 second litters. Toxoplasma gondii was also detected by PCR in 9/30 congenitally infected offspring 16 wk after birth, despite the fact that detectable serological titers had waned. These findings raise questions about the applicability of serological testing to assess the prevalence of T. gondii infection in deer mice and other rodents in the wild. Additionally, the detection of frequent congenital transmission suggests that deer mice could help maintain T. gondii in the environment even in the absence of definitive feline hosts.
Assuntos
Transmissão Vertical de Doenças Infecciosas/veterinária , Peromyscus/parasitologia , Doenças dos Roedores/transmissão , Toxoplasmose Animal/transmissão , Animais , Anticorpos Antiprotozoários/sangue , Reservatórios de Doenças , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Masculino , Reação em Cadeia da Polimerase/veterinária , Doenças dos Roedores/imunologia , Doenças dos Roedores/parasitologia , Toxoplasma/genética , Toxoplasma/imunologia , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/congênito , Toxoplasmose Animal/imunologiaRESUMO
Sarcocystis neurona, a protozoal parasite shed by opossums (Didelphis virginiana), has been shown to cause significant morbidity and mortality in horses, sea otters, and other marine mammals. Over the course of 3 years (fall 2005-summer 2008), opossums from central California were tested for infection with S. neurona. Of 288 opossums sampled, 17 (5.9%) were infected with S. neurona based on the molecular characterization of sporocysts from intestinal scrapings or feces. Risk factors evaluated for association with S. neurona infection in opossums included: age, sex, location, season, presence of pouch young in females, concomitant infection, and sampling method (live-trapped or traffic-killed). Multivariate logistic regression analysis identified that opossums in the Central Valley were 9 times more likely to be infected than those near the coast (p=0.009). Similarly, opossum infection was 5 times more likely to be detected during the reproductive season (March-July; p=0.013). This first investigation of S. neurona infection prevalence and associated risk factors in opossums in the western United States can be used to develop management strategies aimed at reducing the incidence of S. neurona infections in susceptible hosts, including horses and threatened California sea otters (Enhydra lutris neries).
Assuntos
Didelphis/parasitologia , Sarcocystis/fisiologia , Sarcocistose/veterinária , Fatores Etários , Animais , California/epidemiologia , DNA Espaçador Ribossômico/genética , Feminino , Masculino , Análise Multivariada , Prevalência , Fatores de Risco , Sarcocystis/genética , Sarcocistose/epidemiologia , Estações do Ano , Fatores SexuaisRESUMO
Sera from 523 wild rodents were tested for Toxoplasma gondii antibodies using either an indirect fluorescent antibody test (IFAT) (rats and mice, with titer >or=80 considered positive) or a latex agglutination test (LAT) (voles, squirrels, and pocket mice, with titer >or=32 considered positive). Seventeen percent (88/523) of the rodents, including 26% (85/328) of the Peromyscus sp. and 8% (3/37) of Spermophilus beecheyi, were seropositive. Fourteen percent (23/161) of rodents captured in trap sites next to Morro Bay (California) and 15% (16/109) of rodents from sites adjacent to riparian habitats had antibodies to T. gondii, compared to 19% (49/253) of rodents captured in habitats not associated with water; this difference was not statistically significant (P = 0.32). Significantly fewer rodents were captured <200 m from residential housing compared to locations further away (11% vs. 30%, respectively). Factors associated with an increased risk for T. gondii seropositivity in rodents were capture location >or=200 m from residential housing and adult age.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças dos Roedores/epidemiologia , Toxoplasma/imunologia , Toxoplasmose Animal/epidemiologia , Animais , Animais Selvagens , Arvicolinae/parasitologia , California/epidemiologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Testes de Fixação do Látex/veterinária , Modelos Logísticos , Masculino , Camundongos , Peromyscus/parasitologia , Ratos , Fatores de Risco , Doenças dos Roedores/parasitologia , Roedores , Sciuridae/parasitologia , Estudos Soroepidemiológicos , Sigmodontinae/parasitologiaRESUMO
We evaluated the sensitivity (Se) and specificity (Sp) of an IgG enzyme-linked immunosorbent assay (ELISA) and IgG indirect fluorescent antibody test (IFAT) for detection of Toxoplasma gondii-specific antibodies in sera from 2 cat populations using a Bayesian approach. Accounting for test covariance, the Se and Sp of the IgG ELISA were estimated to be 92.6% and 96.5%, and those of the IgG IFAT were 81.0% and 93.8%, respectively. Both tests performed poorly in cats experimentally coinfected with feline immunodeficiency virus and T. gondii. Excluding this group, Se and Sp of the ELISA were virtually unchanged (92.3% and 96.4%, respectively), whereas the IFAT Se improved to 94.2% and Sp remained stable at 93.7%. These tests and an IgM ELISA were applied to 123 cat sera from the Morro Bay area, California, where high morbidity and mortality attributable to toxoplasmosis have been detected in southern sea otters. Age-adjusted IgG seroprevalence in this population was estimated to be 29.6%, and it did not differ between owned and unowned cats. Accounting for Se, Sp, and test covariances, age-adjusted seroprevalence was 45.0%. The odds for T. gondii seropositivity were 12.3-fold higher for cats aged >12 mo compared with cats aged <6 mo.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Gato/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Toxoplasma/imunologia , Toxoplasmose Animal/diagnóstico , Animais , Teorema de Bayes , California/epidemiologia , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , Diagnóstico Diferencial , Fezes/parasitologia , Feminino , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Variações Dependentes do Observador , Fatores de Risco , Estudos Soroepidemiológicos , Organismos Livres de Patógenos Específicos , Toxoplasmose Animal/epidemiologiaRESUMO
The protozoan parasite Toxoplasma gondii is increasingly recognized as a waterborne pathogen. Infection can be acquired by drinking contaminated water and conventional water treatments may not effectively inactivate tough, environmentally resistant oocysts. The present study was performed to assess the efficacy of 2 commonly used chemicals, sodium hypochlorite and ozone, to inactivate T. gondii oocysts in water. Oocysts were exposed to 100 mg/L of chlorine for 30 min, or for 2, 4, 8, 16, and 24 hr, or to 6 mg/L of ozone for 1, 2, 4, 8, or 12 min. Oocyst viability was determined by mouse bioassay. Serology, immunohistochemistry, and in vitro parasite isolation were used to evaluate mice for infection. Initially, mouse bioassay experiments were conducted to compare the analytical sensitivity of these 3 detection methods prior to completing the chemical inactivation experiments. Toxoplasma gondii infection was confirmed by at least 1 of the 3 detection methods in mice inoculated with all doses (10(5)-10(0)) of oocysts. Results of the chemical exposure experiments indicate that neither sodium hypochlorite nor ozone effectively inactivate T. gondii oocysts, even when used at high concentrations.
Assuntos
Desinfetantes/farmacologia , Oxidantes/farmacologia , Ozônio/farmacologia , Hipoclorito de Sódio/farmacologia , Toxoplasma/efeitos dos fármacos , Microbiologia da Água , Animais , Bioensaio , Encéfalo/parasitologia , Gatos , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Oocistos/efeitos dos fármacos , Lontras , Organismos Livres de Patógenos Específicos , Toxoplasmose/prevenção & controle , Toxoplasmose/transmissão , Abastecimento de Água/normasRESUMO
Inactivation of Toxoplasma gondii oocysts occurred with exposure to pulsed and continuous UV radiation, as evidenced by mouse bioassay. Even at doses of >or=500 mJ/cm2, some oocysts retained their viability.
Assuntos
Desinfecção/métodos , Oocistos/efeitos da radiação , Toxoplasma/efeitos da radiação , Raios Ultravioleta , Água/parasitologia , Animais , Gatos , Relação Dose-Resposta à Radiação , Camundongos , Oocistos/crescimento & desenvolvimento , Toxoplasma/crescimento & desenvolvimento , Toxoplasma/patogenicidade , Toxoplasmose/microbiologia , Abastecimento de ÁguaRESUMO
The purpose of this study was to determine the effect of blood contamination of cerebrospinal fluid (CSF) on the results of indirect fluorescent antibody tests (IFATs) for Sarcocystis neurona and Neospora hughesi. The in vitro study used antibody-negative CSF collected from non-neurologic horses immediately after euthanasia and blood samples from 40 healthy horses that had a range of IFAT antibody titers against S. neurona and N. hughesi. Serial dilutions of whole blood were made in seronegative CSF to generate blood-contaminated CSF with red blood cell (RBC) concentrations ranging from 10 to 100,000 RBCs/microl. The blood-contaminated CSF samples were then tested for antibodies against both pathogens using IFAT. Blood contamination of CSF had no detectable effect on IFAT results for S. neurona or N. hughesi at any serologic titer when the RBC concentration in CSF was <10,000 RBCs/microl. At concentrations of 10,000-100,000 RBCs/microl of CSF, positive CSF results (IFAT titer >or=5) for S. neurona and N. hughesi were detected only when the corresponding serum titers were >or=160 and >or=80, respectively. The IFAT performed on CSF is reliable for testing horses for equine protozoal myeloencephalitis caused by S. neurona or N. hughesi, even when blood contamination causes the RBC concentration in CSF to be up to 10,000 RBCs/microl.
Assuntos
Coccidiose/veterinária , Encefalomielite/veterinária , Doenças dos Cavalos/parasitologia , Neospora/isolamento & purificação , Sarcocystis/isolamento & purificação , Sarcocistose/veterinária , Animais , Anticorpos Antiprotozoários/líquido cefalorraquidiano , Coccidiose/sangue , Coccidiose/líquido cefalorraquidiano , Coccidiose/parasitologia , Encefalomielite/sangue , Encefalomielite/líquido cefalorraquidiano , Encefalomielite/parasitologia , Técnica Indireta de Fluorescência para Anticorpo/normas , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Doenças dos Cavalos/sangue , Doenças dos Cavalos/líquido cefalorraquidiano , Doenças dos Cavalos/diagnóstico , Cavalos , Sarcocistose/sangue , Sarcocistose/líquido cefalorraquidiano , Sarcocistose/parasitologia , Manejo de Espécimes/veterináriaRESUMO
Eight female Peromyscus californicus were infected with 10(2) or 10(4) Toxoplasma gondii culture-derived tachyzoites (Type II or X) isolated from southern sea otters. All but 2 mice survived infection and developed antibodies to T. gondii. The 2 fatally infected mice were inoculated with 10(4) tachyzoites of the Type X strain. Parasite detection by immunohistochemistry (IHC) and DNA amplification with 2 polymerase chain reaction (PCR) methods was compared for brain, heart, lung, liver, spleen, biceps muscle, and tongue, at a mean of 41 days postinfection. Parasites were detected most commonly by IHC in spleen (8/8) and brain (6/8). DNA amplification by PCR was most successful from brain, heart, and spleen.
Assuntos
Anticorpos Antiprotozoários/biossíntese , Peromyscus/parasitologia , Doenças dos Roedores/imunologia , Toxoplasma/imunologia , Toxoplasmose Animal/imunologia , Animais , Encéfalo/parasitologia , DNA de Protozoário/análise , Suscetibilidade a Doenças , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Coração/parasitologia , Imuno-Histoquímica , Lontras , Reação em Cadeia da Polimerase , Distribuição Aleatória , Doenças dos Roedores/parasitologia , Doenças dos Roedores/transmissão , Baço/parasitologia , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/transmissãoRESUMO
OBJECTIVE: To estimate risk of exposure and age at first exposure to Sarcocystis neurona and Neospora hughesi and time to maternal antibody decay in foals. ANIMALS: 484 Thoroughbred and Warmblood foals from 4 farms in California. PROCEDURE: Serum was collected before and after colostrum ingestion and at 3-month intervals thereafter. Samples were tested by use of the indirect fluorescent antibody test; cutoff titers were > or = 40 and > or = 160 for S neurona and N hughesi, respectively. RESULTS: Risk of exposure to S neurona and N hughesi during the study were 8.2% and 3.1%, respectively. Annual rate of exposure was 3.1% for S neurona and 1.7% for N hughesi. There was a significant difference in the risk of exposure to S neurona among farms but not in the risk of exposure to N hughesi. Median age at first exposure was 1.2 years for S neurona and 0.8 years for N hughesi. Highest prevalence of antibodies against S neurona and N hughesi was 6% and 2.1 %, respectively, at a mean age of 1.7 and 1.4 years, respectively. Median time to maternal antibody decay was 96 days for S neurona and 91 days for N hughesi. There were no clinical cases of equine protozoal myeloenchaphlitis (EPM). CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to S neurona and N hughesi was low in foals between birth and 2.5 years of age. Maternally acquired antibodies may cause false-positive results for 3 or 4 months after birth, and EPM was a rare clinical disease in horses < or = 2.5 years of age.
Assuntos
Anticorpos Antiprotozoários/sangue , Coccidiose/veterinária , Doenças dos Cavalos/parasitologia , Neospora , Sarcocystis , Fatores Etários , Animais , Anticorpos Antiprotozoários/imunologia , California , Coccidiose/imunologia , Técnica Indireta de Fluorescência para Anticorpo , Doenças dos Cavalos/imunologia , Cavalos , Imunidade Materno-Adquirida/imunologia , Medição de RiscoRESUMO
The objectives of this study were to evaluate the accuracy of the indirect fluorescent antibody test (IFAT) using serum and cerebrospinal fluid (CSF) of horses naturally and experimentally infected with Sarcocystis neurona, to assess the correlation between serum and CSF titers, and to determine the effect of S. neurona vaccination on the diagnosis of infection. Using receiver-operating characteristic analysis, the areas under the curve for the IFAT were 0.97 (serum) and 0.99 (CSF). Sensitivity and specificity were 83.3 and 96.9% (serum, cutoff 80) and 100 and 99% (CSF, cutoff 5), respectively. Titer-specific likelihood ratios (LRs) ranged from 0.03 to 187.8 for titers between <10 and 640. Median time to conversion was 22-26 days postinfection (DPI) (serum) and 30 DPI (CSF). The correlation between serum and CSF titers was moderately strong (r = 0.6) at 30 DPI. Percentage of vaccinated antibody-positive horses ranged from 0 to 95% between 0 and 112 days after the second vaccination. Thus, the IFAT was reliable and accurate using serum and CSF. Use of LRs potentially improves clinical decision making. Correlation between serum and CSF titers affects the joint accuracy of the IFAT; therefore, the ratio of serum to CSF titers has potential diagnostic value. The S. neurona vaccine could possibly interfere with equine protozoal myeloencephalitis diagnosis.
Assuntos
Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/líquido cefalorraquidiano , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Doenças dos Cavalos/imunologia , Sarcocystis/imunologia , Sarcocistose/veterinária , Animais , Encefalomielite/diagnóstico , Encefalomielite/parasitologia , Encefalomielite/veterinária , Feminino , Doenças dos Cavalos/sangue , Doenças dos Cavalos/líquido cefalorraquidiano , Cavalos , Funções Verossimilhança , Masculino , Vacinas Protozoárias/imunologia , Curva ROC , Reprodutibilidade dos Testes , Sarcocistose/sangue , Sarcocistose/líquido cefalorraquidiano , Sarcocistose/imunologia , Sensibilidade e Especificidade , Vacinação/veterináriaRESUMO
The study objective was to assess the risk of transplacental transmission of Sarcocystis neurona and Neospora hughesi in foals from 4 California farms during 3 foaling seasons. Serum of presuckle foals and serum and colostrum of periparturient mares were tested using indirect fluorescent antibody tests for S. neurona and N. hughesi. Serum antibody titers were < or =10 in 366 presuckle foals tested. There was no serologic or histologic evidence of either parasite in aborted fetuses or placentas examined. Positivity for S. neurona and N. hughesi in mares increased with age. Mares < or =9 yr that originated from Kentucky were 3.8 and 1.4 times more likely to be positive for S. neurona and N. hughesi, respectively, than mares from California. The strength of association between positivity to either parasite and state of birth decreased as age increased. Mares positive for S. neurona and N. hughesi were 2.2 and 1.7 times more likely, respectively, to have a previous abortion than negative mares, adjusted for age and state of birth. The annual mortality rate for mares was 4%. The annual incidence rate of equine protozoal myeloencephalitis was 0.2%. In conclusion, there was no detectable risk of transplacental transmission of S. neurona and N. hughesi. Prevalence of antibodies against both parasites in mares increased with age.