RESUMEN
A flagellated enteric diplomonad protozoan consistent with Spironucleus meleagridis (formerly Hexamita meleagridis) associated with gastrointestinal disease and mortality in psittacine birds including cockatiels (Nymphicus hollandicus) has been sporadically described in the literature. However, molecular characterization of psittacine protozoal isolates had not yet been performed. The 16S rRNA gene from a protozoan persistently shed in the feces in a small group of cockatiels demonstrated a 98% molecular identity with S. meleagridis isolated from turkeys. Based on these sequence data, a diagnostic PCR assay was developed to detect the presence of S. meleagridis. Nineteen privately owned pet cockatiels from unrelated households were clinically evaluated. All birds microscopically positive for this organism were PCR positive, with several additional birds microscopically negative but PCR positive. Many of the birds identified as positive for S. meleagridis by fecal PCR had signs of gastrointestinal disease such as diarrhea, soft feces, and melena, whereas none of the birds that tested negative had gastrointestinal signs. Examination of feces from two unrelated cockatiel breeding facilities revealed 70% and 86% PCR positive rates. Prevalence of infection and incidence of clinical disease, including factors that lead to clinical manifestation such as viral, bacterial, or mycotic coinfections, are not yet known and warrant further study, but spironucleosis is likely an under-recognized disease in cockatiels.
Asunto(s)
Enfermedades de las Aves/parasitología , Cacatúas/parasitología , Diplomonadida/aislamiento & purificación , Infecciones Protozoarias en Animales/parasitología , Animales , Diplomonadida/genética , Heces/parasitología , Femenino , Masculino , Reacción en Cadena de la Polimerasa/métodosRESUMEN
Bartonella species are gram-negative bacteria that infect erythrocytes, endothelial cells and macrophages, often leading to persistent blood-borne infections. Because of the ability of various Bartonella species to reside within erythrocytes of a diverse number of animal hosts, there is substantial opportunity for the potential uptake of these blood-borne bacteria by a variety of arthropod vectors that feed on animals and people. Five Bartonella species are transmitted by lice, fleas or sandflies. However, Bartonella DNA has been detected or Bartonella spp. have been cultured from numerous other arthropods. This review discusses Bartonella transmission by sandflies, lice and fleas, the potential for transmission by other vectors, and data supporting transmission by ticks. Polymerase chain reaction (PCR) or culture methods have been used to detect Bartonella in ticks, either questing or host-attached, throughout the world. Case studies and serological or molecular surveys involving humans, cats and canines provide indirect evidence supporting transmission of Bartonella species by ticks. Of potential clinical relevance, many studies have proposed co-transmission of Bartonella with other known tick-borne pathogens. Currently, critically important experimental transmission studies have not been performed for Bartonella transmission by many potential arthropod vectors, including ticks.
Asunto(s)
Vectores Arácnidos/microbiología , Infecciones por Bartonella/transmisión , Infecciones por Bartonella/veterinaria , Garrapatas/microbiología , Zoonosis , Animales , Bartonella/aislamiento & purificación , ADN Bacteriano/química , ADN Bacteriano/genética , Reservorios de Enfermedades , Humanos , Insectos Vectores/microbiología , Phthiraptera/microbiología , Siphonaptera/microbiologíaRESUMEN
BACKGROUND: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. HYPOTHESIS: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. ANIMALS: Ninety-five dogs and 52 cats presented during a neutering campaign. METHODS: A prospective cross-sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. RESULTS: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66-67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.
Asunto(s)
Enfermedades de los Gatos/epidemiología , Enfermedades Transmisibles/veterinaria , Enfermedades de los Perros/epidemiología , Animales , Gatos , Enfermedades Transmisibles/epidemiología , Estudios Transversales , Perros , Ecuador/epidemiología , Enfermedades Endémicas/veterinaria , Femenino , Masculino , Prevalencia , Estudios ProspectivosRESUMEN
Trematodes belonging to the genus Bolbophorus have recently been reported as the cause of substantial morbidity and mortality in cultured channel catfish Ictalurus punctatus in Mississippi and Louisiana. Previous investigators identified only a single species, B. confusus. In this investigation, genetic techniques were used to identify all stages of the parasite in all of its hosts. The 18s rRNA genes from specimens collected in Mississippi were sequenced and compared; this analysis revealed that there are two distinct species, B. damnificus (previously identified as B. confusus) and another, undescribed species. (Phylogenetic analysis indicated that a third species, B. levantinus, is also closely related to the Mississippi species.) Species-specific polymerase chain reaction assays capable of identifying and differentiating between these two parasites were developed. Both species were found to infect the first intermediate host (the ram's horn snail Planorbella trivolvis) in commercial channel catfish ponds, but only B. damnificus was recovered from the fish themselves. The new, unidentified Bolbophorus species was determined to be highly pathogenic to a number of fish species. The contribution of B. damnificus to disease in cultured channel catfish remains undetermined. Future investigations of these parasites must now take into account the presence of two distinct species.
RESUMEN
Babesia canis has generally been considered the only large Babesia to infect dogs. Here we describe the molecular characterization of a large Babesia species that was detected in the blood and bone marrow of a dog with clinical and hematological abnormalities consistent with babesiosis. Analysis of the 18S rRNA genes revealed a unique sequence that shared 93.9% sequence identity with B. bigemina and 93.5% sequence identity with B. caballi, compared to 91.2-91.6% identity with B. canis canis, B. c. vogeli, and B. c. rossi. Cross-reactive antibodies against B. canis, B. gibsoni (Asian genotype), or B. gibsoni (California genotype) antigens were not detected in acute or convalescent serum samples. The dog was treated with imidocarb diproprionate, which resulted in the resolution of clinical signs, and subsequently Babesia DNA was not detectable by PCR in post-treatment samples. The organism described in this report represents a genetically unique large Babesia sp. and is the eighth genetically distinct piroplasm capable of infecting the domestic dog.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Babesia/clasificación , Babesia/aislamiento & purificación , Babesiosis/veterinaria , Enfermedades de los Perros/parasitología , Animales , Antiprotozoarios/uso terapéutico , Babesia/genética , Babesia/inmunología , Babesiosis/tratamiento farmacológico , Babesiosis/parasitología , Reacciones Cruzadas , Enfermedades de los Perros/tratamiento farmacológico , Perros , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Imidocarbo/uso terapéutico , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , ARN Ribosómico 18S , Homología de Secuencia de Ácido Nucleico , Resultado del TratamientoRESUMEN
Infections by trematodes are among the most common fish-borne zoonoses. Metacercariae of the Family Heterophyidae in marine and freshwater fishes are nonfastidious in their choice of definitive hosts, and therefore, cause infections in human and domestic animals. In the present study, species-specific polymerase chain reaction (PCR) assays were developed for identifying and differentiating the various species examined. Sequencing and aligning the 18S (SSU) rDNA revealed interspecific variation for which species-specific DNA oligonucleotides were designed and used for the identification of 6 heterophyid species recovered from piscivorous birds. The oligonucleotides were further used to evaluate the various stages (cercariae recovered from snails, metacercariae recovered from fish and adult trematodes) of the digeneans. By applying this method we elucidated for the first time the life cycle of Pygidiopsis genata. The phylogenetic interrelationship among the newly sequenced species of Heterophyidae is outlined.
Asunto(s)
Aves/parasitología , Peces/parasitología , Heterophyidae/genética , Estadios del Ciclo de Vida/genética , Filogenia , Caracoles/parasitología , Animales , Secuencia de Bases , Análisis por Conglomerados , Cartilla de ADN , ADN Ribosómico/genética , Electroforesis en Gel de Agar , Agua Dulce , Funciones de Verosimilitud , Modelos Genéticos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Infections by metacercariae of Clinostomum (Leidy, 1856) species adversely affect aquacultured fish and are potentially transmissible to humans. Molecular methodologies are efficient tools, which enable diagnosis of all life-history stages of trematodes in their diverse hosts. The small subunit of ribosomal DNA genes of adults of the Old World Clinostomum complanatum (Rudolphi, 1819) and the New World Clinostomum marginatum (Rudolphi, 1819), obtained from a little egret Egretta garzetta (Linnaeus, 1766) and the great blue heron Ardea herodias (Linnaeus, 1758), respectively, were amplified, sequenced, and aligned. The resulting alignment was used to develop a genetic assay to differentiate between these species.
Asunto(s)
Enfermedades de las Aves/parasitología , ADN Ribosómico/genética , Enfermedades de los Peces/parasitología , Trematodos/clasificación , Infecciones por Trematodos/veterinaria , Animales , Secuencia de Bases , Aves , Cyprinidae/parasitología , Cartilla de ADN/química , ADN de Helmintos/química , ADN de Helmintos/genética , ADN Ribosómico/química , Enfermedades de los Peces/transmisión , Variación Genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/veterinaria , Polimorfismo Genético , Alineación de Secuencia/veterinaria , Especificidad de la Especie , Trematodos/genética , Infecciones por Trematodos/parasitologíaRESUMEN
Metacercariae of Bolbophorus species are serious pathogens of farmed fish. Molecular diagnostic tools, capable of identifying and differentiating these parasites, may assist in the development of rationale control strategies. The rDNA 18S (small sub-unit: SSU) genes of adult B. confusus and B. levantinus obtained from a pelican, Pelecanus onocrotalus, and a night heron, Nycticorax nycticorax, respectively, were amplified, sequenced, and aligned. Based on this alignment, we developed a genetic differentiation assay between B. confusus and B. levantinus. These 2 species were compared genetically with the North American species B. damnificus and Bolbophorus sp. ('Type 2'). The relationship between species is outlined and discussed. In addition to the molecular study, specimens of B. confusus and B. levantinus were compared morphologically, using scanning electron microscopy. Morphologic analysis revealed interspecific differences in details of the holdfast organ and the position of the acetabulum.
Asunto(s)
Variación Genética , Filogenia , Platelmintos/genética , Platelmintos/ultraestructura , Animales , Secuencia de Bases , Análisis por Conglomerados , Cartilla de ADN , ADN Ribosómico/genética , Funciones de Verosimilitud , Microscopía Electrónica de Rastreo , Modelos Genéticos , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Trematodes belonging to the genus Bolbophorus have recently been reported as the cause of substantial morbidity and mortality in cultured channel catfish Ictalurus punctatus in Mississippi and Louisiana. Previous investigators identified only a single species, B. confusus. In this investigation, genetic techniques were used to identify all stages of the parasite in all of its hosts. The 18s rRNA genes from specimens collected in Mississippi were sequenced and compared; this analysis revealed that there are two distinct species, B. damnificus (previously identified as B. confusus) and another, undescribed species. (Phylogenetic analysis indicated that a third species, B. levantinus, is also closely related to the Mississippi species.) Species-specific polymerase chain reaction assays capable of identifying and differentiating between these two parasites were developed. Both species were found to infect the first intermediate host (the ram's horn snail Planorbella trivolvis) in commercial channel catfish ponds, but only B. damnificus was recovered from the fish themselves. The new, unidentified Bolbophorus species was determined to be highly pathogenic to a number of fish species. The contribution of B. damnificus to disease in cultured channel catfish remains undetermined. Future investigations of these parasites must now take into account the presence of two distinct species.
RESUMEN
OBJECTIVE: To determine whether infection with Tritrichomonas foetus causes diarrhea in specific-pathogen-free or Cryptosporidium coinfected cats. ANIMALS: 4 cats with subclinical cryptosporidiosis (group 1) and 4 specific-pathogen-free cats (group 2). PROCEDURE: Cats were infected orogastrically with an axenic culture of T. foetus isolated from a kitten with diarrhea. Direct microscopy and protozoal culture of feces, fecal character, serial colonic mucosal biopsy specimens, and response to treatment with nitazoxanide (NTZ; group 1) or prednisolone (groups 1 and 2) were assessed. RESULTS: Infection with T. foetus persisted in all cats for the entire 203-day study and resulted in diarrhea that resolved after 7 weeks. Group-1 cats had an earlier onset, more severe diarrhea, and increased number of trichomonads on direct fecal examination, compared with group-2 cats. Use of NTZ eliminated shedding of T. foetus and Cryptosporidium oocysts, but diarrhea consisting of trichomonad-containing feces recurred when treatment was discontinued. Prednisolone did not have an effect on infection with T. foetus but resulted in reappearance of Cryptosporidium oocysts in the feces of 2 of 4 cats. During necropsy, T. foetus was isolated from contents of the ileum, cecum, and colon. Tritrichomonas foetus organisms and antigen were detected on surface epithelia and within superficial detritus of the cecal and colonic mucosa. CONCLUSIONS AND CLINICAL RELEVANCE: After experimental inoculation in cats, T. foetus organisms colonize the ileum, cecum, and colon, reside in close contact with the epithelium, and are associated with transient diarrhea that is exacerbated by coexisting cryptosporidiosis but not treatment with prednisolone.
Asunto(s)
Enfermedades de los Gatos/parasitología , Enfermedades Intestinales/veterinaria , Infecciones Protozoarias en Animales , Tritrichomonas foetus , Animales , Antígenos de Protozoos/análisis , Antiprotozoarios/administración & dosificación , Antiprotozoarios/uso terapéutico , Enfermedades de los Gatos/tratamiento farmacológico , Gatos , Criptosporidiosis/complicaciones , Criptosporidiosis/tratamiento farmacológico , Criptosporidiosis/veterinaria , Cryptosporidium/crecimiento & desarrollo , Diarrea/tratamiento farmacológico , Diarrea/parasitología , Diarrea/veterinaria , Heces/parasitología , Femenino , Enfermedades Intestinales/tratamiento farmacológico , Enfermedades Intestinales/parasitología , Mucosa Intestinal/parasitología , Nitrocompuestos , Recuento de Huevos de Parásitos/veterinaria , Prednisolona/administración & dosificación , Prednisolona/uso terapéutico , Infecciones por Protozoos/complicaciones , Infecciones por Protozoos/tratamiento farmacológico , Organismos Libres de Patógenos Específicos , Tiazoles/administración & dosificación , Tiazoles/uso terapéuticoAsunto(s)
Enfermedades de los Gatos/parasitología , Diarrea/veterinaria , Infecciones Protozoarias en Animales , Tritrichomonas foetus , Animales , Enfermedades de los Gatos/diagnóstico , Gatos , Diagnóstico Diferencial , Diarrea/diagnóstico , Diarrea/parasitología , Heces/parasitología , Giardia/clasificación , Giardiasis/diagnóstico , Giardiasis/veterinaria , Infecciones por Protozoos/diagnóstico , Tritrichomonas foetus/clasificaciónRESUMEN
Canine ehrlichiosis is a highly variable syndrome presenting a significant differential diagnostic difficulty. It imitates many metabolic and infectious diseases and lacks standardized diagnostic criteria, common reagents, and database resources. A clinical diagnosis of canine ehrlichiosis may be based on the manifestation of fever, thrombocytopenia, anorexia, nasolacrimal discharge, epistaxis, and exclusion of autoimmune and common canine vector borne diseases. These parameters are not invariably observed especially in the atypical form of the disease often caused by species other than Ehrlichia canis. A definitive diagnosis is based on the presence of specific antibodies to ehrlichial agent(s), the demonstration of the etiologic agent(s) itself, or specific amplicons by a strigently quality controlled PCR protocol. The relationship of the various clinical and laboratory parameters, the status of the currently available tests, and their real or presumed predictive value are discussed in the context of stimulating an effort to formulate an international standard for the diagnosis of this and related diseases of man and animals.
Asunto(s)
Enfermedades de los Perros/diagnóstico , Ehrlichiosis/veterinaria , Animales , Anorexia , Diagnóstico Diferencial , Enfermedades de los Perros/fisiopatología , Perros , Ehrlichia/clasificación , Ehrlichia/aislamiento & purificación , Ehrlichiosis/diagnóstico , Ehrlichiosis/fisiopatología , Epistaxis , Fiebre , Masculino , Reacción en Cadena de la Polimerasa , TrombocitopeniaRESUMEN
OBJECTIVE: To establish clinical features, course of illness, and treatment outcome of cats with diarrhea and concurrent infection with Trichomonas organisms. Prevalence of fecal trichomonads in a geographically comparable population of healthy indoor and feral cats also was assessed. DESIGN: Longitudinal study and a cohort study. ANIMALS: 32 cats with diarrhea and naturally acquired trichomonosis that were native to North Carolina, Virginia, Connecticut, and Tennessee; 20 healthy indoor cats; and 100 feral cats. PROCEDURE: Trichomonosis was diagnosed in 32 cats by identification of organisms in fresh feces or by protozoal culture of feces. RESULTS: Diarrhea associated with the large intestine and trichomonosis were diagnosed in 32 cats. Median age of the cats was 9 months; 23 cats were < or = 1 year old at the time of diagnosis. Two cats developed diarrhea accompanied by infection with Trichomonas organisms after the addition of an infected kitten into the home. Duration of diarrhea ranged from 2 days to 3 years. Six cats had a coexisting enteric infection. Treatment with antimicrobials improved fecal consistency and reduced the number of flagellates in the feces, but did not eliminate infection. Diarrhea (with microscopically detectable flagellates) was observed shortly after antibiotics were discontinued. Trichomonads were not recovered from feces of any healthy indoor or feral cats. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings suggest that trichomonosis may be a cofactor in development of diarrhea in young cats. Trichomonas organisms were not identified as part of the indiginous fauna of healthy indoor or feral cats.
Asunto(s)
Enfermedades de los Gatos/epidemiología , Diarrea/veterinaria , Parasitosis Intestinales/veterinaria , Tricomoniasis/veterinaria , Animales , Animales Domésticos , Animales Salvajes , Enfermedades de los Gatos/parasitología , Gatos , Estudios de Cohortes , Connecticut/epidemiología , Diarrea/complicaciones , Diarrea/epidemiología , Heces/parasitología , Femenino , Parasitosis Intestinales/epidemiología , Intestino Grueso/parasitología , Estudios Longitudinales , Masculino , North Carolina/epidemiología , Prevalencia , Tennessee/epidemiología , Tricomoniasis/complicaciones , Tricomoniasis/epidemiología , Virginia/epidemiologíaRESUMEN
The recognition of canine babesiosis in North Carolina caused by Babesia gibsoni documents the expansion of the previously reported endemic area of this disease. Clinical signs ranged from severe hemolytic anemia and thrombocytopenia to subclinical infections. No infected dogs had traveled to endemic areas. Antibabesial treatment failed to eradicate the organism from infected dogs.
Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/epidemiología , Enfermedades de los Perros/epidemiología , Animales , Babesiosis/diagnóstico , Babesiosis/terapia , Diagnóstico Diferencial , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/microbiología , Perros , Femenino , Masculino , North Carolina/epidemiologíaRESUMEN
The dinoflagellate Amyloodinium ocellatum, which causes amyloodiniosis or 'marine velvet disease', is one of the most serious ectoparasitic diseases plaguing warmwater marine fish culture worldwide. We report that tomato clownfish Amphiprion frenatus develop strong immunity to Amyloodinium ocellatum infection following repeated nonlethal challenges and that specific antibodies are associated with this response. Reaction of immune fish antisera against dinospore and trophont-derived antigens in Western blots indicated both shared and stage-specific antibody-antigen reactions. A mannan-binding-protein affinity column was used to isolate IgM-like antibody from A. frenatus serum. The reduced Ig consisted of one 70 kD heavy chain and one 32 kD light chain with an estimated molecular weight of 816 kD for the native molecule. Immunoglobulin (Ig) isolated from immune but not non-immune fish serum significantly inhibited parasite infectivity in vitro. An enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal rabbit antibody produced against affinity-purified A. frenatus Ig. Anti-Amyloodinium serum antibody was not always detectable in immune fish, although serum antibody titers in immune fish increased after repeated exposure to the parasite. These results suggest that there may be a localized antibody response in skin/gill epithelial tissue, although antibody was rarely detected in skin mucus.
Asunto(s)
Anticuerpos Antiprotozoarios/biosíntesis , Dinoflagelados/inmunología , Enfermedades de los Peces/inmunología , Infecciones Protozoarias en Animales , Animales , Anticuerpos Antiprotozoarios/sangre , Especificidad de Anticuerpos , Antígenos de Protozoos/inmunología , Western Blotting , Cromatografía de Afinidad/veterinaria , Relación Dosis-Respuesta Inmunológica , Electroforesis en Gel de Poliacrilamida/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedades de los Peces/prevención & control , Peces , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Sueros Inmunes/inmunología , Inmunización Pasiva , Inmunodifusión/veterinaria , Inmunoglobulinas/sangre , Infecciones por Protozoos/inmunología , Infecciones por Protozoos/prevención & controlRESUMEN
Three antibacterial proteins were isolated from acid extracts of channel catfish (Ictalurus punctatus) skin by cation exchange chromatography and reverse-phase high-pressure liquid chromatography. The molecular masses of the proteins were 15.5, 15.5 and 30 kD as determined by SDS-polyacrylamide gel electrophoresis. Mass spectrometry, amino acid composition and amino acid sequence data suggest that the most abundant protein is closely related to histone H2B. The H2B-like protein was inhibitory to Aeromonas hydrophila and Saprolegnia spp., which are important bacterial and fungal pathogens of fish. These findings suggest that histones may be important defensive molecules in fish.
Asunto(s)
Antibacterianos/aislamiento & purificación , Histonas/química , Ictaluridae/inmunología , Proteínas/aislamiento & purificación , Piel/química , Aeromonas/efectos de los fármacos , Secuencia de Aminoácidos , Animales , Antibacterianos/farmacología , Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Electroforesis en Gel de Poliacrilamida , Enfermedades de los Peces/inmunología , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Peso Molecular , Oomicetos/efectos de los fármacos , Proteínas/farmacología , Piel/microbiologíaRESUMEN
Historically, considerable variation has been reported in the type and severity of clinical and hematologic abnormalities associated with canine ehrlichiosis. Because of difficulties associated with the isolation of intracellular monocytic Ehrlichia species in tissue culture systems, few E. canis isolates are available for comparative microbiologic studies. To address the issue of potential E. canis antigenic diversity in different regions of the world, dog sera reactive by indirect fluorescent antibody testing to E. canis (Florida) antigen were obtained from France, Israel, Italy, the United States, the Virgin Islands, and Zimbabwe. Ehrlichia canis proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and at least 5 sera from each region were stained by western immunoblotting. Antibody immunodominance was scored based upon staining intensity. There was relative homogeneity in the immunogenic protein reactions to E. canis antigens. Of the 58 E. canis reactive sera, 54 samples resulted in immunoblot patterns indicative of chronic ehrlichiosis. Four reactive sera (reciprocal titers of 160-2,560) did not recognize any genus-specific antigens resulting in protein bands between 22 and 29 kD, indicating serologic cross-reactivity with other microorganisms. Relatively homogenous immunoblot patterns, consistent with the reported immunoblot response of dogs with experimental chronic ehrlichiosis, were observed with sera from Arizona, France, Israel, North Carolina, Texas, and the Virgin Islands. In contrast, unique major proteins were observed in dog sera from Italy and Zimbabwe. Our results indicate that although relatively homogeneous, antigenic diversity may exist among E. canis organisms in different regions of the world.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Enfermedades de los Perros , Ehrlichia/inmunología , Ehrlichiosis/veterinaria , Inmunoglobulina G/sangre , Animales , Formación de Anticuerpos , Antígenos Bacterianos/inmunología , Western Blotting/métodos , Perros , Ehrlichia/genética , Ehrlichiosis/inmunología , Francia , Variación Genética , Israel , Italia , Estados Unidos , Islas Virgenes de los Estados Unidos , ZimbabweRESUMEN
A simple and reproducible method was developed for the measurement of blastogenesis of peripheral blood lymphocytes using whole blood of hybrid bass (striped bass [Morone saxatilis] female x white bass [M. chrysops] male) stimulated with Concanavalin A, phytohemagglutinin-P, lipopolysaccharide or pokeweed mitogen. Compared to traditional methods which use leucocyte separation procedures, whole blood culture is faster and less expensive. Only small aliquots of blood (10 microliters per culture well) were needed, which would be beneficial for sampling small fish as well as for taking multiple samples from single animals. Optimal culture conditions for hybrid bass, including mitogen concentration, incubation temperature and incubation period, were determined. This is the first report to demonstrate a blastogenic response of whole blood cells in fish.
Asunto(s)
Lubina/inmunología , Activación de Linfocitos , Animales , Femenino , Masculino , Mitógenos/farmacología , TemperaturaRESUMEN
A polymerase chain reaction (PCR) for amplifying ribosomal DNA of Rickettsia rickettsii was performed on blood clots and urine samples from 10 patients with suspected Rocky Mountain spotted fever (RMSF) and five controls with nonrickettsial diseases. The results of this PCR-based procedure were positive in four of the five patients with probable RMSF, but reamplification was required in three patients. Rickettsia rickettsii was grown from the blood of two of these four patients. The urine from one patient was also PCR-positive. These results confirm earlier findings that the PCR can detect R. rickettsii, but the need for reamplification indicates that the lack of sensitivity is a serious limitation in the usefulness of the PCR as a clinical diagnostic test.
Asunto(s)
ADN Bacteriano/análisis , ADN Ribosómico/análisis , Reacción en Cadena de la Polimerasa , Rickettsia rickettsii/genética , Fiebre Maculosa de las Montañas Rocosas/diagnóstico , Bacteriemia/microbiología , Bacteriuria/microbiología , ADN Bacteriano/sangre , ADN Bacteriano/orina , ADN Ribosómico/sangre , ADN Ribosómico/orina , Electroforesis en Gel de Agar , Femenino , Humanos , Masculino , Estudios Prospectivos , ARN Ribosómico 16S/genética , Rickettsia rickettsii/aislamiento & purificación , Sensibilidad y EspecificidadRESUMEN
The ability of tabanid mouthparts to retain and to transfer mechanically Ehrlichia risticii Holland, Weiss, Burgdorfer, Cole & Kakoma was evaluated by feeding flies on infected and noninfected mice and on capillary tubes containing infected cells and cell-free medium. Flies representing two genera and 29 species were collected at equine boarding stables, farms, and along riding trails in Wake, Johnston, and Duplin counties in North Carolina for the feeding trials. Two species, Tabanus fulvulus Wiedemann and T. pallidescens Philip, fed on mice but failed to transfer the pathogen from infected to susceptible mice. Chrysops vittatus Wiedemann, Tabanus americanus Forster, and T. sulcifrons Macquart transferred E. risticii-infected cells from capillary tubes containing infected cells in medium to tubes containing medium. These studies document that E. risticii-infected cells can be retained on mouthparts and potentially transferred by tabanids.