RESUMO
Horses are prone to recurrent airway obstruction (RAO), an inflammatory lung disease induced by repeated exposure to environmental mold, dust, and bacterial components. Active disease manifests with mucus hyperproduction, neutrophilic inflammation, bronchoconstriction, and coughing. Chronically affected animals have lung remodeling characterized by smooth muscle hyperplasia, collagen deposition, lymphoid hyperplasia, and impaired aerobic performance. Clara cell secretory protein (CCSP) counters inflammation in the lung, hence we hypothesized that CCSP depletion is a key feature of RAO in horses. Recombinant equine CCSP and specific antiserum were produced, and percutaneous lung biopsies were obtained from 3 healthy horses and from 3 RAO-affected horses before and after induction of RAO. CCSP relative gene expression in tissue, as well as protein concentration in lung lavage fluid, was determined. Immunocytochemical analysis, using both light and immunogold ultrastructural methods, demonstrated reduced CCSP staining in lung tissue of animals with RAO. Immunogold label in Clara cell granules was less in animals with chronic RAO than in normal animals, and absent in animals that had active disease. Median lung lavage CCSP concentration was 132 and 129 ng/ml in healthy horses, and 62 and 24 ng/ml in RAO horses before and after challenge, respectively. CCSP lung gene expression was significantly higher in healthy animals than in animals with chronic RAO. Together, these preliminary findings suggest that reduced production of CCSP and subcellular changes in Clara cells are features of chronic environmentally induced lung inflammation in horses.
Assuntos
Obstrução das Vias Respiratórias/veterinária , Regulação da Expressão Gênica/fisiologia , Doenças dos Cavalos/metabolismo , Uteroglobina/metabolismo , Obstrução das Vias Respiratórias/metabolismo , Animais , Lavagem Broncoalveolar/veterinária , Clonagem Molecular , Primers do DNA/genética , Ensaio de Imunoadsorção Enzimática , Cavalos , Imuno-Histoquímica/veterinária , Pulmão/metabolismo , Pulmão/ultraestrutura , Microscopia Eletrônica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Uteroglobina/genéticaRESUMO
A critical period of early gestation in the mare involves the immobilization (fixation) of the encapsulated conceptus at around days 16-17. We compared the major proteins in the normal equine embryonic capsule and endometrial secretions around the period of fixation with those from pregnancies in the process of termination induced by administration of an analogue of prostaglandin F(2 alpha) (PGF(2 alpha)). Uterocalin and beta(2)-microglobulin (beta(2)M) associated with the embryonic capsule were proteolytically converted to smaller forms during the fixation period. These conversions were similar in conceptuses from control and treated mares. A 17 kDa cationic protein identified as a secretory phospholipase A2 (sPLA2) type IIA was detected bound to normal capsules but increased substantially in response to PGF(2 alpha). Two forms of uteroglobin were distinguished by partial amino acid sequences of approximately 6 kDa bands in flush fluids from normal pregnant uteri. After administration of PGF(2 alpha) one immunoreactive form of uteroglobin was preferentially increased. These studies demonstrate that failure of pregnancy in this model is associated with an increase in secretory phospholipase in the capsule and a change in the forms of uteroglobin in the uterine secretions.
Assuntos
Implantação do Embrião/fisiologia , Embrião de Mamíferos/metabolismo , Glicoproteínas/metabolismo , Cavalos/fisiologia , Prenhez/fisiologia , Animais , Eletroforese em Gel de Poliacrilamida , Feminino , Idade Gestacional , Glicoproteínas/análise , Cavalos/metabolismo , Gravidez , Prenhez/metabolismo , Uteroglobina/análise , Uteroglobina/metabolismo , Útero/química , Útero/metabolismo , Saco Vitelino/química , Saco Vitelino/metabolismo , Microglobulina beta-2/análise , Microglobulina beta-2/metabolismoRESUMO
BACKGROUND: Lead is a persistent contaminant in the environment, and waterfowl are susceptible to lead toxicity from ingestion of lead pellets and fishing weights. Lead affects numerous physiologic processes through inhibition of enzyme activity and protein function, but its effects on commonly assessed avian blood values are incompletely understood. OBJECTIVES: Our aim was to evaluate hematologic and biochemical changes associated with blood lead concentrations in trumpeter swans and Canada geese. METHODS: Data for CBCs, plasma biochemical profiles (total protein, albumin, glucose, cholesterol, total bilirubin, calcium, phosphorus, gamma-glutamyltransferase [GGT], aspartate aminotransferase, lactate dehydrogenase, glutamate dehydrogenase, creatine kinase, amylase, and lipase), and whole blood lead concentrations were retrospectively analyzed for 69 trumpeter swans and 52 Canada geese. Laboratory data obtained prospectively from an additional 20 trumpeter swans also were included. RBC morphology was semiquantitated in blood smears from 70 of the birds. Data were analyzed initially by ANOVA and covariance. A statistical model then was constructed to determine the relationship between each parameter and lead concentration. RESULTS: In both avian species, PCV, hemoglobin concentration, and MCHC decreased significantly (P < .05) with increasing blood lead concentration. Uric acid concentration and GGT activity were increased in trumpeter swans and phosphorus concentration was decreased in Canada geese in association with high blood lead concentration (P < .05). CONCLUSIONS: Lead toxicosis induced significant changes in the values of commonly measured hematologic parameters in waterfowl. These changes may be useful indicators of severe lead intoxication during routine laboratory assessment. Changes in clinical chemistry values, although statistically significant, were too inconsistent to serve as indicators of lead toxicosis.
Assuntos
Anseriformes/sangue , Anseriformes/metabolismo , Doenças das Aves/sangue , Doenças das Aves/metabolismo , Intoxicação por Chumbo/veterinária , Animais , Animais Selvagens , Doenças das Aves/patologia , Intoxicação por Chumbo/sangue , Intoxicação por Chumbo/metabolismo , Intoxicação por Chumbo/patologia , Estudos Prospectivos , Estudos RetrospectivosRESUMO
To determine whether the presence of nonpathogenic piroplasms may confound field estimates of risk of Babesia microti infection, we identified sporozoites infecting the salivary glands of deer ticks (Ixodes dammini) by parallel microscopy and polymerase chain reaction assays. Piroplasms were evident in 14.4% of adult ticks from sites in the northcentral and northeastern United States. Of these, 83.3% contained DNA characteristic of Ba. odocoilei. This cervid piroplasm was detected in all of the sites examined and generally was more prevalent than was Ba. microti. Because deer ticks transmit both Ba. odocoilei and Ba. microti, estimates of pathogen prevalence based solely on microscopy may overestimate the risk of human babesiosis.
Assuntos
Vetores Aracnídeos/parasitologia , Babesia/classificação , Ixodes/parasitologia , Animais , Babesia/genética , Babesia/isolamento & purificação , Sequência de Bases , DNA de Protozoário/análise , DNA de Protozoário/química , DNA Ribossômico/química , Cervos/parasitologia , Feminino , Maine , Massachusetts , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 18S/genética , Coelhos , Alinhamento de Sequência , Análise de Sequência de DNA , WisconsinRESUMO
Interleukin-4 has been reported to critically modulate Borrelia burgdorferi infection and Lyme arthritis in experimental murine models. To determine the in vivo role of IL-4 in controlling Lyme carditis, we compared immunological responses and the severity of cardiac inflammation in wild-type BALB/c (IL-4 +/+) and IL-4 deficient BALB/c (IL-4 -/-) mice infected with B. burgdorferi by tick-bite. At day 15 and 30 post-infection IL-4 -/- mice produced significantly greater titers of spirochete-specific IgG2a than the wild-type IL-4 +/+ mice, which produced significantly more spirochete-specific IgG1. Following in vitro antigenic stimulation with B. burgdorferi antigen, splenocytes from infected IL-4 -/- and IL-4 +/+ mice displayed similar magnitudes of proliferative responses at day 15 and 30 post-infection. At day 30 antigen-stimulated splenocytes from infected IL-4 -/- mice, however, produced significantly more IFN-gamma than those derived from similarly infected IL-4 +/+ mice, suggesting that Th1-influenced responses predominated in IL-4 -/- mice. Moreover, inflamed hearts from IL-4 -/- mice displayed higher levels of IFN-gamma and TNF-alpha transcripts as compared to IL-4 +/+ mice. At both time points antigen-stimulated splenocytes from IL-4 +/+ and IL-4 -/- mice produced significant amounts of IL-10 but those from IL-4 +/+ mice produced either no or little IL-4. Histopathology demonstrated typical Lyme carditis in both IL-4 +/+ and IL-4 -/- mice at day 15 and day 30. Although Borrelia-infected IL-4 -/- mice developed a more severe carditis on day 30, the carditis resolved by day 50, as it did in IL4 +/+ mice. These results indicate that although IL-4 may help limit the severity of Lyme carditis, its absence does not preclude resolution of cardiac lesions.
Assuntos
Interleucina-4/fisiologia , Doença de Lyme/imunologia , Miocardite/imunologia , Células Th1/imunologia , Animais , Anticorpos Antibacterianos/sangue , Imunoglobulina G/sangue , Imunoglobulina G/classificação , Interferon gama/biossíntese , Interleucina-10/biossíntese , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Dogs were found to be susceptible to human granulocytotropic Ehrlichia spp. Infection was produced through the bite of Ixodes scapularis Say (= dammini Spielman, Clifford, Piesman & Corwin) nymphs and adults that acquired infection while feeding as larvae on experimentally infected mice. Dogs were also infected by intravenous injection of mouse blood or dog blood from parasitemic donors. Parasites were demonstrable in neutrophils within 8 or 9 d after nymphs began feeding; prepatent periods were longer when infection was induced by adult tick feeding (18 d) or by transfusion of mouse blood (12 d). The shortest prepatent period observed was 5 d in a dog infected by transfusion of blood from a parasitemic dog. Infections in dogs were mild and apparently transient. Mild thrombocytopenia was the most commonly observed abnormality. Parasites could be detected by light microscopy during the acute phase of infection (4 or 5 d) and parasite DNA by polymerase chain reaction as early as 5 d after exposure but not at 6-9 d after morulae were first observed in neutrophils. Likewise, dog blood was infectious for mice at 2 d but not at 25 d, and for dogs at 3 d but not at 13 d after morulae were first observed in neutrophils. Seroconversion occurred as early as 11 d after onset of tick feeding and persisted until dogs were euthanatized. Gross and histopathologic lesions were similar to those observed in dogs with E. canis (Donatien & Lestoquard), E. chaffeensis Anderson, Dawson & Wilson, and E. ewingii Anderson, Greene, Jones & Dawson infections but were generally milder than any of these. The moderate enlargement of lymphoid organs observed grossly was reflected histologically as mild to moderate reactive hyperplasia, which was largely follicular (B cell).
Assuntos
Reservatórios de Doenças , Doenças do Cão/microbiologia , Ehrlichiose/veterinária , Granulócitos/microbiologia , Animais , Vetores Aracnídeos/microbiologia , Doenças do Cão/transmissão , Cães , Ehrlichia/isolamento & purificação , Ehrlichiose/microbiologia , Ehrlichiose/transmissão , Células HL-60 , Humanos , Ixodes/microbiologia , Camundongos , Camundongos Endogâmicos C3HRESUMO
Horses suffer from recurrent airway obstruction, an asthma-like condition induced by repeat inhalation of environmental substances present in barn air. Clara cell secretory protein (CCSP) is much reduced during active inflammation when neutrophils predominate in the airways, and in chronic asthmatics. We sought to investigate morphologic and functional interactions of CCSP with neutrophils. Bronchoalveolar and blood neutrophils from healthy control animals, and from animals with recurrent airway obstruction in remission and exacerbation, were evaluated by immuno-cytochemistry and immuno-electron microscopy for presence of CCSP. Blood neutrophil oxidative burst and phagocytic activities were determined in the presence of different concentrations of recombinant equine CCSP. Bronchoalveolar lavage neutrophils from horses with exacerbated lung inflammation, but not from control horses, and not blood neutrophils from either group of animal, contained abundant immunoreactive CCSP. On immuno-electron microscopy, CCSP localized to the cytoplasm and nucleus. Incubation of blood neutrophils with CCSP significantly reduced oxidative burst activity (P<0.0001) and increased phagocytosis (P<0.001) of neutrophils. These findings indicate that CCSP enters neutrophils in horses with active neutrophilic lung inflammation and alters the function of neutrophils in blood. Presence in the nucleus suggests a potential transcriptional role of CCSP in neutrophils.
Assuntos
Neutrófilos/fisiologia , Oxirredução/efeitos dos fármacos , Fagocitose/fisiologia , Uteroglobina/fisiologia , Obstrução das Vias Respiratórias/imunologia , Obstrução das Vias Respiratórias/fisiopatologia , Obstrução das Vias Respiratórias/veterinária , Animais , Líquido da Lavagem Broncoalveolar/citologia , Citometria de Fluxo/veterinária , Doenças dos Cavalos/imunologia , Doenças dos Cavalos/fisiopatologia , Cavalos/metabolismo , Cavalos/fisiologia , Microscopia Imunoeletrônica/veterinária , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Fagocitose/efeitos dos fármacos , Uteroglobina/farmacologiaRESUMO
Deer tick-transmitted pathogens such as Lyme disease spirochetes and babesiae appear to require a period of reactivation and replication during the tick's blood meal before it is able to infect a host. The duration of nymphal tick attachment that is required for transmission of the agent of human granulocytic ehrlichiosis (HGE) was determined by removing feeding ticks from mice at various time points. As with spirochetes and babesiae, ehrlichiae infected few mice when ticks were removed prior to 36 h of tick attachment. This "grace period" may serve as a modifying factor in the epidemiology of this newly emergent zoonosis and help physicians make informed decisions concerning management of tick bites in HGE-endemic areas.
Assuntos
Ehrlichiose/transmissão , Granulócitos/microbiologia , Carrapatos , Animais , Cervos/parasitologia , Ehrlichia/isolamento & purificação , Ehrlichia/fisiologia , Ehrlichiose/sangue , Humanos , Estágios do Ciclo de Vida , Camundongos , Camundongos Endogâmicos C3H , Glândulas Salivares/parasitologia , Fatores de TempoRESUMO
To determine if eastern North American Ixodes dammini, like related ticks in Eurasia, maintain tick-borne encephalitis group viruses, we analyzed ticks collected from sites where the agent of Lyme disease is zoonotic. Two viral isolates were obtained by inoculating mice with homogenates from tick salivary glands. The virus, which was described by reverse transcriptase polymerase chain reaction and direct sequencing of the amplification products, was similar to, but distinct from, Powassan virus and is provisionally named "deer tick virus." Enzootic tick-borne encephalitis group viruses accompany the agents of Lyme disease, babesiosis, and granulocytic ehrlichiosis in a Holarctic assemblage of emergent deer tick pathogens.
Assuntos
Vírus da Encefalite/isolamento & purificação , Ixodes/virologia , Animais , Vírus da Encefalite/classificação , Vírus da Encefalite/patogenicidade , Camundongos , Reação em Cadeia da PolimeraseRESUMO
A human-derived strain of the agent of human granulocytic ehrlichiosis, a recently described emerging rickettsial disease, has been established by serial blood passage in mouse hosts. Larval deer ticks acquired infection by feeding upon such mice and efficiently transmitted the ehrlichiae after molting to nymphs, thereby demonstrating vector competence. The agent was detected by demonstrating Feulgen-positive inclusions in the salivary glands of the experimentally infected ticks and from field-derived adult deer ticks. White-footed mice from a field site infected laboratory-reared ticks with the agent of human granulocytic ehrlichiosis, suggesting that these rodents serve as reservoirs for ehrlichiae as well as for Lyme disease spirochetes and the piroplasm that causes human babesiosis. About 10% of host-seeking deer ticks were infected with ehrlichiae, and of these, 20% also contained spirochetes. Cotransmission of diverse pathogens by the aggressively human-biting deer tick may have a unique impact on public health in certain endemic sites.
Assuntos
Cervos/microbiologia , Ehrlichia/isolamento & purificação , Ehrlichiose/microbiologia , Carrapatos/metabolismo , Animais , Vetores Aracnídeos , Sequência de Bases , Cricetinae , DNA Bacteriano , Ehrlichia/patogenicidade , Ehrlichiose/transmissão , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos DBA , Dados de Sequência MolecularRESUMO
We investigated whether Ixodes scapularis-mediated host immunity interrupts transmission of the agent of human granulocytic ehrlichiosis (aoHGE) to guinea pigs. Ticks infected with aoHGE readily transmitted aoHGE to tick-immune guinea pigs, despite incomplete tick engorgement and host attachment. Although tick immunity can prevent Lyme borreliosis, protection is not afforded against granulocytic ehrlichiosis.
Assuntos
Ehrlichiose/prevenção & controle , Ixodes/imunologia , Animais , Ehrlichiose/transmissão , Feminino , Cobaias , Células HL-60 , Humanos , Immunoblotting , Camundongos , Camundongos Endogâmicos C3H , Camundongos SCID , Reação em Cadeia da PolimeraseRESUMO
C3H mice that were inoculated with ehrlichiae isolated from a patient with human granulocytic ehrlichiosis (HGE) developed anemia and leukopenia, but by day 24, they returned to normal values. Granulocytic morulae were present in peripheral blood and spleen smears on days 5 and 10, and there was a reduction in morulae on day 17. Ehrlichiae were present in HL-60 cell cultures of blood and spleen from all mice at all intervals. Pathogenicity, but not infectivity, waned with mouse passage but could be resurrected by SCID mouse passage. Various methods were tested for their relative sensitivity in detecting infection: blood smears, HL-60 cell cultures, polymerase chain reaction (PCR) amplification of a 16S recombinant DNA target, and a mouse infectivity assay. All assays detected the HGE agent in blood during early infection, but PCR and the mouse infectivity assay were most sensitive during late infection. Xenodiagnosis demonstrated that mice remain persistently infected through 55 days.