RESUMO
Algumas espécies de Staphylococcus causam infecções crônicas intramamárias e podem levar à formação de biofilme. No presente estudo, levantou-se a hipótese de que as espécies de Staphylococcus isolados da mastite bovina são capazes de formar biofilme in vitro associado à presença dos genes icaA, icaD ou bap. Um total de 200 isolados de Staphylococcus, sendo 100 Staphylococcus aureus de casos de mastite subclínica e 100 estafilococos não aureus (ENA) de casos de mastite subclínica e clínica, obtidos em duas fazendas leiteiras, no estado de São Paulo, foram avaliados quanto à capacidade de produzir biofilmes in vitro. A presença de icaA, icaD e bap foi confirmada por PCR, e a produção de biofilme em ágar vermelho congo (Congo Red Agar - CRA) e em teste de microplaca (Microtiter Plate - MtP) foi avaliada nos isolados de S. aureus e ENA. Os resultados mostraram a presença dos genes icaA, icaD e bap em S. aureus, mas não em ENA. A produção de biofilme pode estar associada à presença de outros fatores ou genes que estimulam a produção de biofilme in vitro. O ensaio de MtP serve como um modelo quantitativo para o estudo da aderência de espécies de estafilococos associados à mastite bovina.(AU)
Assuntos
Animais , Feminino , Bovinos , Staphylococcus/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Biofilmes , Mastite Bovina/diagnóstico , Reação em Cadeia da Polimerase/veterinária , ÁgarRESUMO
Algumas espécies de Staphylococcus causam infecções crônicas intramamárias e podem levar à formação de biofilme. No presente estudo, levantou-se a hipótese de que as espécies de Staphylococcus isolados da mastite bovina são capazes de formar biofilme in vitro associado à presença dos genes icaA, icaD ou bap. Um total de 200 isolados de Staphylococcus, sendo 100 Staphylococcus aureus de casos de mastite subclínica e 100 estafilococos não aureus (ENA) de casos de mastite subclínica e clínica, obtidos em duas fazendas leiteiras, no estado de São Paulo, foram avaliados quanto à capacidade de produzir biofilmes in vitro. A presença de icaA, icaD e bap foi confirmada por PCR, e a produção de biofilme em ágar vermelho congo (Congo Red Agar - CRA) e em teste de microplaca (Microtiter Plate - MtP) foi avaliada nos isolados de S. aureus e ENA. Os resultados mostraram a presença dos genes icaA, icaD e bap em S. aureus, mas não em ENA. A produção de biofilme pode estar associada à presença de outros fatores ou genes que estimulam a produção de biofilme in vitro. O ensaio de MtP serve como um modelo quantitativo para o estudo da aderência de espécies de estafilococos associados à mastite bovina.(AU)
Assuntos
Animais , Feminino , Bovinos , Staphylococcus/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Biofilmes , Mastite Bovina/diagnóstico , Reação em Cadeia da Polimerase/veterinária , ÁgarRESUMO
AIMS: To investigate the anti-inflammatory activity of an invasive and Hp65-producing strain Lactococcus lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) in acute 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced colitis in mice as an innovative therapeutic strategy against Crohn's disease (CD). METHODS AND RESULTS: The pXYCYT:Hsp65 plasmid was transformed into the L. lactis NCDO2118 FnBPA+ strain, resulting in the L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) strain. Then, the functionality of the strain was evaluated in vitro for Hsp65 production by Western blotting and for invasion into Caco-2 cells. The results demonstrated that the strain was able to produce Hsp65 and efficiently invade eukaryotic cells. Subsequently, in vivo, the anti-inflammatory capacity of the recombinant strain was evaluated in colitis induced with TNBS in BALB/c mice. Oral administration of the recombinant strain was able to attenuated the severity of colitis by mainly reducing IL-12 and IL-17 levels and increasing IL-10 and secretory immunoglobulin A levels. CONCLUSIONS: The L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) strain contributed to a reduction in inflammatory damage in experimental CD. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, which used L. lactis for the production and delivery of Hsp65, has scientific relevance because it shows the efficacy of this new strategy based on therapeutic protein delivery into mammalian enterocytes.
Assuntos
Proteínas de Bactérias/metabolismo , Chaperonina 60/metabolismo , Colite/terapia , Imunoglobulina A Secretora/metabolismo , Interleucina-10/metabolismo , Lactococcus lactis/fisiologia , Administração Oral , Animais , Proteínas de Bactérias/genética , Células CACO-2 , Chaperonina 60/genética , Colite/induzido quimicamente , Colite/imunologia , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Inflamação/terapia , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ácido Trinitrobenzenossulfônico/toxicidadeRESUMO
Oral mucositis (OM) is a common and dose-limiting side effect of cancer treatment, including 5-fluorouracil (5-FU) and radiotherapy. The efficacy of the therapeutic measures to prevent OM is limited and disease prevention is not fully observable. Amifostine is a cytoprotective agent with a described anti-inflammatory potential. It is clinically used to reduce radiotherapy and chemotherapy-associated xerostomia. This study investigated the protective effect of amifostine on an experimental model of OM. Hamsters were divided into six groups: saline control group (5 mL/kg), mechanical trauma (scratches) of the right cheek pouch; 5-FU (60 and 40 mg/kg, ip, respectively, administered on days 1 and 2); amifostine (12.5, 25, or 50 mg/kg) + 5-FU + scratches. Salivation rate was assessed and the animals were euthanized on day 10 for the analysis of macroscopic and microscopic injury by scores. Tissue samples were harvested for the measurement of neutrophil infiltration and detection of inflammatory markers by ELISA and immunohistochemistry. 5-FU induced pronounced hyposalivation, which was prevented by amifostine (P<0.05). In addition, 5-FU injection caused pronounced tissue injury accompanied by increased neutrophil accumulation, tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1ß) tissue levels, and positive immunostaining for TNF-α, IL-1ß, and inducible nitric oxide synthase (iNOS). Interestingly, amifostine prevented the inflammatory reaction and consequently improved macroscopic and microscopic damage (P<0.05 vs 5-FU group). Amifostine reduced inflammation and protected against 5-FU-associated oral mucositis and hyposalivation.
Assuntos
Amifostina/uso terapêutico , Fluoruracila/efeitos adversos , Inflamação/prevenção & controle , Substâncias Protetoras/uso terapêutico , Estomatite/prevenção & controle , Xerostomia/prevenção & controle , Animais , Cricetinae , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Inflamação/patologia , Masculino , Estomatite/induzido quimicamente , Estomatite/patologia , Xerostomia/induzido quimicamente , Xerostomia/patologiaRESUMO
Oral mucositis (OM) is a common and dose-limiting side effect of cancer treatment, including 5-fluorouracil (5-FU) and radiotherapy. The efficacy of the therapeutic measures to prevent OM is limited and disease prevention is not fully observable. Amifostine is a cytoprotective agent with a described anti-inflammatory potential. It is clinically used to reduce radiotherapy and chemotherapy-associated xerostomia. This study investigated the protective effect of amifostine on an experimental model of OM. Hamsters were divided into six groups: saline control group (5 mL/kg), mechanical trauma (scratches) of the right cheek pouch; 5-FU (60 and 40 mg/kg, ip, respectively, administered on days 1 and 2); amifostine (12.5, 25, or 50 mg/kg) + 5-FU + scratches. Salivation rate was assessed and the animals were euthanized on day 10 for the analysis of macroscopic and microscopic injury by scores. Tissue samples were harvested for the measurement of neutrophil infiltration and detection of inflammatory markers by ELISA and immunohistochemistry. 5-FU induced pronounced hyposalivation, which was prevented by amifostine (P<0.05). In addition, 5-FU injection caused pronounced tissue injury accompanied by increased neutrophil accumulation, tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) tissue levels, and positive immunostaining for TNF-α, IL-1β, and inducible nitric oxide synthase (iNOS). Interestingly, amifostine prevented the inflammatory reaction and consequently improved macroscopic and microscopic damage (P<0.05 vs 5-FU group). Amifostine reduced inflammation and protected against 5-FU-associated oral mucositis and hyposalivation.
Assuntos
Animais , Masculino , Estomatite/prevenção & controle , Xerostomia/prevenção & controle , Amifostina/uso terapêutico , Substâncias Protetoras/uso terapêutico , Fluoruracila/efeitos adversos , Inflamação/prevenção & controle , Estomatite/induzido quimicamente , Estomatite/patologia , Xerostomia/induzido quimicamente , Xerostomia/patologia , Cricetinae , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Inflamação/patologiaRESUMO
AIMS: A regimen utilizing Bacille Calmette-Guerin (BCG) and another vaccine system as a booster may represent a promising strategy for the development of an efficient tuberculosis vaccine for adults. In a previous work, we confirmed the ability of Lactococcus lactis fibronectin-binding protein A (FnBPA+) (pValac:ESAT-6), a live mucosal DNA vaccine, to produce a specific immune response in mice after oral immunization. In this study, we examined the immunogenicity of this strain as a booster for the BCG vaccine in mice. METHODS AND RESULTS: After immunization, cytokine and immunoglobulin profiles were measured. The BCG prime L. lactis FnBPA+ (pValac:ESAT-6) boost group was the most responsive group, with a significant increase in splenic pro-inflammatory cytokines IL-17, IFN-γ, IL-6 and TNF-α compared with the negative control. CONCLUSIONS: Based on the results obtained here, we demonstrated that L. lactis FnBPA+ (pValac:ESAT-6) was able to increase the BCG vaccine general immune response. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is of great scientific and social importance because it represents the first step towards the development of a booster to the BCG vaccine using L. lactis as a DNA delivery system.
Assuntos
Antígenos de Bactérias/imunologia , Vacina BCG/imunologia , Proteínas de Bactérias/imunologia , Citocinas/sangue , Interleucina-17/metabolismo , Lactococcus lactis/genética , Vacinas de DNA/imunologia , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Feminino , Interleucina-6 , Lactococcus lactis/metabolismo , Camundongos , Fator de Necrose Tumoral alfaRESUMO
The purpose of the study was to compare the peripheral retinal sensitivity of the visual field between professional soccer players and age-gender matched non-athlete subjects. All participants underwent a complete eye evaluation. The visual field was evaluated with the achromatic program 60-4 from the Humphrey automated perimetry. The binocular visual field was created with the best location model. It was divided into 4 quadrants (left superior, right superior, left inferior, and right inferior) and compared between groups. The study group comprised 29 professional male football players and the control group comprised 26 age-matched male non-athletes. Mean age was 25.8±4.7 years in the study group and 26.3±5.1 for controls. The average of retina sensitivity in the left inferior and right inferior quadrants was higher in the study group (27.2±1.2 dB and 27.0±1.4 dB) as compared to controls (26.1±1.9 dB and 25.5±2.1 dB). (Student's t test, P=0.011 and P=0.004, respectively). In this small cohort, professional soccer players presented higher retina sensitivity in the inferior quadrants when compared to non-athletes.
Assuntos
Atletas , Retina/fisiologia , Futebol , Campos Visuais/fisiologia , Adolescente , Adulto , Estudos Transversais , Humanos , Masculino , Testes de Campo Visual , Adulto JovemRESUMO
In this study, Lactococcus lactis was engineered to express mutated internalin A on its surface and to secrete large amounts of listeriolysin O (LLO) in order to improve its potential as a vehicle for DNA vaccination. Western blotting experiments demonstrated that the bacterium expressed LLO in both the cytoplasmic and extracellular compartments, with higher quantities found in the culture supernatants. A hemolytic assay showed that the recombinant strain secreted 250 ng active LLO/mg total protein. This mInlA/LLO-producing strain of L. lactis may be used as an alternative tool in DNA vaccination against a number of infectious diseases or in cancer therapy.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Toxinas Bacterianas/genética , Toxinas Bacterianas/imunologia , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/imunologia , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/imunologia , Lactococcus lactis/genética , Listeria monocytogenes/imunologia , Mutação , Proteínas Recombinantes , Vacinas Bacterianas , Membrana Celular/metabolismo , Expressão Gênica , Hemólise , Lactococcus lactis/metabolismo , VacinaçãoRESUMO
Toxoplasmosis is the major parasitic disease affecting sheep. It is important for veterinary medicine, animal science and public health since it causes reproductive and economic losses in the herd, as well as damaging human health due to consumption of contaminated meat and milk, which can facilitate zoonotic transmission. Detection of Toxoplasma gondii in ovine milk and lack of data in the literature describing differentiation between acute and chronic disease for this species stimulated the elaboration of the present research project. To achieve the aim of this study, the animals were allocated to two groups of 20 ewes each, of which group 1 was composed of animals with positive serology and group 2 with negative serology. Acute and chronic stages of the disease were differentiated by modified direct agglutination test (MAT), in which antigens were fixed with formalin (MAT-AF) and methanol (MAT-AM). The parasite was detected in milk by polymerase chain reaction (PCR), and the molecular identity of the amplified products was confirmed by sequencing. The serological results indicated that sheep had a chronic infection profile. T. gondii DNA was detected in seven milk samples from five seropositive sheep, and twice in milk of two sheep. Sequences of species shared 97-100% identity with T. gondii. These findings allowed the hypothesis that the peripartum period may also lead to the resurgence of tissue T. gondii tachyzoites cysts which can circulate again and be excreted in the milk. This study used sheep naturally infected with T. gondii as a prerequisite for further investigations on the possible participation of this species in toxoplasmosis epidemiology and as a potential transmission route related to consumption of milk from infected sheep.
Assuntos
DNA de Protozoário/análise , Leite/parasitologia , Doenças dos Ovinos/parasitologia , Toxoplasma/genética , Toxoplasmose Animal/parasitologia , Zoonoses/parasitologia , Testes de Aglutinação/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Brasil/epidemiologia , DNA de Protozoário/química , DNA de Protozoário/genética , Feminino , Reação em Cadeia da Polimerase/veterinária , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/imunologia , Toxoplasmose Animal/sangue , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/imunologia , Zoonoses/epidemiologia , Zoonoses/transmissãoRESUMO
Road-killed wild animals have been for years used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies. In the current study, fungal infection was evaluated by PCR and nested-PCR in tissue samples collected from 19 road-killed wild animals. The necropsies were carried out and samples were collected for DNA extraction. Results, using PCR with a panfungal primer and nested PCR with specific primers, indicated that some animals are naturally infected with Amauroascus aureus, Metarhizium anisopliae, Aspergillus flavus, Aspergillus oryzae, Emmonsia parva, Paracoccidioides brasiliensis or Pichia stipitis. The approach employed herein proved useful for detecting the environmental occurrence of several fungi, as well as determining natural reservoirs in wild animals and facilitating the understanding of host-pathogen relationships.(AU)