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1.
J Dairy Sci ; 103(6): 5376-5386, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32229113

RESUMO

We analyzed a large number of immune response parameters from quarter milk samples with distinct bacteriological and quarter somatic cell count (qSCC) statuses. Furthermore, we sought to explore and identify displayed immune response patterns in milk samples from mammary glands with nonspecific mastitis. Thus, 92 quarter milk samples from 28 cows were stratified into 4 groups, as follows: (1) 49 culture-negative control quarters with a low qSCC (<1 × 105 cells/mL) from 19 dairy cows (so-called healthy quarters); (2) 15 culture-negative quarters with high qSCC (>2 × 105 cells/mL; so-called quarters with nonspecific mastitis) from 10 dairy cows; (3) 8 culture-positive quarters with low qSCC (noninflammatory quarters with low qSCC) from 5 dairy cows; and (4) 20 culture-positive quarters with high qSCC (so-called truly infected quarters) from 8 dairy cows. Using flow cytometry, we evaluated the percentage of milk neutrophils and their viability, intracellular reactive oxygen species production, phagocytosis, and the expression of CD62L, CD11b, and CD44 for each of the 4 quarter strata. Furthermore, the percentage of monocyte/macrophages, B cells, and T lymphocyte subsets were evaluated by flow cytometry. Milk samples from bacteriologically negative quarters (both with a low and elevated qSCC) had a lower qSCC than those with bacteriologically positive outcomes (both with a low and elevated qSCC). As expected, the healthy quarters showed the lowest percentage of neutrophils and also showed a higher percentage of milk monocytes/macrophages and lower percentage of T lymphocytes than truly infected quarters. The most prominent result of the present study is that quarters with nonspecific mastitis showed the highest percentage of milk CD4+ T lymphocytes. The healthy quarters had a lower percentage of apoptotic neutrophils than noninflammatory and truly infected quarters, although it did not differ from those from the quarters with nonspecific mastitis. Our study supports the role of differential cell counting in the diagnosis of mastitis, as the milk leukocyte populations markedly fluctuate under healthy and inflammatory conditions. Furthermore, an increase in milk CD4+ T cells was associated with nonspecific mastitis, suggesting an increase in this leukocyte subpopulation is correlated with low bacterial shedding. Our study allows us to go further in our understanding of mammary gland immunity, providing further insights on potential protective mammary gland immunity, which we hypothesize can open new avenues for the development of novel targets that can promote bovine udder health.


Assuntos
Mastite Bovina/imunologia , Leite , Animais , Linfócitos B , Bovinos , Contagem de Células/veterinária , Feminino , Citometria de Fluxo/veterinária , Contagem de Linfócitos , Glândulas Mamárias Animais/imunologia , Glândulas Mamárias Animais/microbiologia , Mastite Bovina/diagnóstico , Mastite Bovina/microbiologia , Leite/citologia , Leite/microbiologia , Neutrófilos , Fagocitose , Linfócitos T
2.
J Dairy Sci ; 103(5): 4588-4605, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32113759

RESUMO

Staphylococcus aureus is one of the pathogens most frequently isolated from cases of mastitis worldwide. To decrease the effect of S. aureus mastitis in dairy farming, alternative strategies for controlling mastitis are needed that depend on a better knowledge of cow-to-cow variations in S. aureus antibody production. The present study sought to explore the diversity of S. aureus antibodies produced by dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine. We obtained protein extracts from S. aureus isolates derived from persistent subclinical mastitis. Proteins were fractionated using 2-dimensional gel electrophoresis and Western blotting. Then, Western blotting membranes were exposed to sera from 24 dairy cows that had been divided into the following groups: vaccinated dairy cows that were infected with S. aureus, further subdivided according to whether they (a) remained infected by S. aureus or (b) recovered from the intramammary infection; unvaccinated dairy cows infected with S. aureus; and vaccinated healthy dairy cows with no history of S. aureus mastitis. Proteins found to be reactive by Western blot were identified by mass spectrometry (MALDI/TOF-TOF). Our most important finding was that F0F1 ATP synthase subunit α, succinyl-diaminopimelate desuccinylase, and cysteinyl-tRNA synthetase were potential candidate proteins for the prevention of S. aureus mastitis. This study strengthens the notion that variations among animals should not be ignored and shows that the heterogeneity of antibody production against anti-staphylococcal antigens in animals may enable the identification of new immunotherapy targets.


Assuntos
Anticorpos Antibacterianos/sangue , Mastite Bovina/imunologia , Infecções Estafilocócicas/veterinária , Vacinas Antiestafilocócicas/administração & dosagem , Staphylococcus aureus/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Bovinos , Feminino , Humanos , Mastite Bovina/microbiologia , Mastite Bovina/prevenção & controle , Leite , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Vacinas Antiestafilocócicas/imunologia
3.
Invest New Drugs ; 37(2): 282-290, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30101387

RESUMO

Background Preclinical studies suggest that imatinib resistance in gastrointestinal stromal tumor (GIST) can be mediated by MAP-kinase activation via fibroblast growth factor (FGF) signaling. In FGF stimulated GIST cell lines, BGJ398, a pan-FGFR kinase inhibitor in combination with imatinib, was cytotoxic and superior to imatinib therapy alone. In FGF-dependent GIST, the combination of BGJ398 and imatinib may provide a mechanism to overcome imatinib resistance. Methods This phase Ib study of BGJ398 and imatinib was performed in patients with imatinib refractory advanced GIST. A standard 3 + 3 dosing schema was utilized to determine the recommended phase II dose (RP2D). Two treatment schedules were evaluated incorporating imatinib 400 mg daily in combination with (A) BGJ398 daily 3 weeks on, 1 week off or (B) BGJ398 daily 1 week on, 3 weeks off. Results 16 patients enrolled. The median age was 54 years (range: 44-77), 81% were male, and the median number of lines of prior therapy was 4 [range: 2-6, 13 patients had ≥3 prior therapies]. 12 patients received treatment on schedule A [BGJ398 dose range: 25 - 75 mg]: 2 patients experienced dose limiting toxicities (DLT) (n = 1, myocardial infarction & grade (G)4 CPK elevation; n = 1, G3 ALT elevation) on schedule A (BGJ398 75 mg), significant hyperphosphatemia, an on-target effect, was not observed, implying the maximum tolerated dose was below the therapeutic dose. Following protocol amendment, 4 patients enrolled on schedule B [BGJ398 dose range: 75 - 100 mg]: no DLTs were observed. The most common treatment related adverse events occurring in >15% of patients included CPK elevation (50%), lipase elevation (44%), hyperphosphatemia (24%), anemia (19%), and peripheral edema (19%). Among the 12 evaluable patients, stable disease (SD) was the best response observed in 7 patients by RECIST v1.1 and 9 patients by CHOI. Stable disease ≥ 32 weeks was observed in 3 patients (25%). Median progression free survival was 12.1 weeks (95% CI 4.7-19.5 weeks). Conclusions Toxicity was encountered with the combination therapy of BGJ398 and imatinib. Due to withdrawal of sponsor support the study closed before the RP2D or dosing schedule of the combination therapy was identified. In heavily pre-treated patients, stable disease ≥ 32 weeks was observed in 3 of 12 evaluable patients. Trial Registration: NCT02257541 .


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Gastrointestinais/tratamento farmacológico , Tumores do Estroma Gastrointestinal/tratamento farmacológico , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inibidores de Proteínas Quinases/administração & dosagem , Receptores de Fatores de Crescimento de Fibroblastos/antagonistas & inibidores , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/farmacocinética , Biomarcadores Tumorais/metabolismo , Feminino , Seguimentos , Neoplasias Gastrointestinais/enzimologia , Neoplasias Gastrointestinais/secundário , Tumores do Estroma Gastrointestinal/enzimologia , Tumores do Estroma Gastrointestinal/patologia , Humanos , Mesilato de Imatinib/administração & dosagem , Masculino , Dose Máxima Tolerável , Pessoa de Meia-Idade , Metástase Neoplásica , Compostos de Fenilureia/administração & dosagem , Prognóstico , Pirimidinas/administração & dosagem , Distribuição Tecidual
4.
World J Surg Oncol ; 17(1): 27, 2019 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728011

RESUMO

BACKGROUND: Few studies of robotic nipple sparing mastectomy (NSM) were reported. We report feasibility of robotic NSM and determine standard surgical procedure and learning curve threefold. METHODS: A cohort of patients with robotic NSM for breast cancer was analyzed. Complications and post-operative hospitalization stay were reported. The same technic was used for all patients except for skin and nipple areolar complex (NAC) dissection. Differences between three surgical procedures of NAC dissection were analyzed: group 1, dissection with robotic scissors using coagulation; group 2, dissection with robotic scissors without coagulation; and group 3, dissection with non-robotic scissors and then robotic dissection. We explored possible effect of learning curve among patients from group 1 with the same surgical procedure. RESULTS: Twenty-seven NSM with immediate breast reconstruction for breast cancers, 22 invasive and 5 in situ, were performed, with robotic latissimus dorsi-flap (RLDF) only in 17 cases, RLDF and breast implant in 6 cases, and implant alone in 4 cases. Repartition according to 3 surgical procedure groups was 16, 5, and 6 patients. Mean time of surgery and anesthesia decrease according to groups 1 to 3. Among 16 patients from group 1, time of surgery and anesthesia decreased with learning curve. Post-operative hospitalization decreased from group 1 to 3. We reported a total of 11 complications, with significant difference between groups (10 for group 1). Skin complications were higher for group 1 in comparison with groups 2-3 (p = 0.02). CONCLUSION: Robotic NSM can be performed with a brief learning. Standardized technique is proposed with non-robotic scissors superficial dissection and then dissection with robot.


Assuntos
Neoplasias da Mama/cirurgia , Mastectomia Subcutânea/efeitos adversos , Mamilos , Tratamentos com Preservação do Órgão/efeitos adversos , Complicações Pós-Operatórias/epidemiologia , Procedimentos Cirúrgicos Robóticos/efeitos adversos , Adolescente , Adulto , Idoso , Estudos de Coortes , Dissecação/efeitos adversos , Dissecação/educação , Dissecação/métodos , Estudos de Viabilidade , Feminino , Humanos , Curva de Aprendizado , Tempo de Internação/estatística & dados numéricos , Mastectomia Subcutânea/educação , Mastectomia Subcutânea/instrumentação , Mastectomia Subcutânea/métodos , Pessoa de Meia-Idade , Duração da Cirurgia , Tratamentos com Preservação do Órgão/instrumentação , Tratamentos com Preservação do Órgão/métodos , Complicações Pós-Operatórias/etiologia , Procedimentos Cirúrgicos Robóticos/educação , Procedimentos Cirúrgicos Robóticos/instrumentação , Procedimentos Cirúrgicos Robóticos/métodos , Resultado do Tratamento , Adulto Jovem
5.
Internist (Berl) ; 60(7): 756-760, 2019 Jul.
Artigo em Alemão | MEDLINE | ID: mdl-30980098

RESUMO

This article reports the case of a 26-year-old male patient with recurrent emesis and headache due to central nervous system tuberculosis. The thoracic computed tomography showed bilateral disseminated pulmonary micronodular infiltrates and a cavern connecting to the bronchial system. The cranial magnetic resonance imaging showed multiple supratentorial and infratentorial microabscesses with concomitant meningitis. Mycobacterium tuberculosis was detected in sputum, bronchoalveolar lavage and cerebrospinal fluid. The patient received first-line antituberculous drug treatment, including streptomycin (instead of ethambutol) and adjuvant prednisolone.


Assuntos
Cefaleia/etiologia , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose/diagnóstico , Vômito/etiologia , Adulto , Antituberculosos/uso terapêutico , Lavagem Broncoalveolar , Líquido Cefalorraquidiano/microbiologia , Etambutol/uso terapêutico , Humanos , Imageamento por Ressonância Magnética , Masculino , Escarro/microbiologia , Tomografia Computadorizada por Raios X , Tuberculose/tratamento farmacológico , Tuberculose/microbiologia
6.
Microb Pathog ; 123: 264-268, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30040999

RESUMO

Hobi-like viruses (HobiPeV) comprise a novel, recently classified species of bovine pestiviruses, originally identified in commercial fetal bovine serum of Brazilian origin and, subsequently, isolated from diseased animals in several countries. Although frequently isolated from clinical cases, most HobiPeV isolates failed to reproduce overt disease in cattle upon experimental inoculation. Herein, we describe the outcome of experimental infection of four to six months-old seronegative calves with two Brazilian HobiPeV isolates. Calves inoculated intranasally with isolate SV478/07 developed viremia between days 2 and 9 post-inoculation (pi) and shed virus in nasal secretions up to day 11pi. These animals presented hyperthermia (day 7 to 10-11 pi) and lymphopenia from days 4 to 8pi. Clinically, all four calves developed varied degrees of apathy, anorexia, mild to moderate respiratory signs (nasal secretion, hyperemia), ocular discharge and pasty diarrhea in the days following virus inoculation. In contrast, calves inoculated with isolate SV757/15 presented only hyperthermia (days 3 to 10-11 pi) and lymphopenia (days 4-8 pi), without other apparent clinical signs. In these animals, viremia was detected up to day 9 pi and virus shedding in nasal secretions lasted up to day 12-14 pi. Both groups seroconverted to the inoculated viruses, developing virus neutralizing (VN) titers from 320 to 5120 at day 28pi. These results extend previous findings that experimental infections of calves with HobiPeV are predominantly mild, yet they also indicate that field isolates may differ in their ability to cause disease in susceptible animals.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Doenças dos Bovinos/virologia , Bovinos/virologia , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/patogenicidade , Febre/virologia , Linfopenia/virologia , Infecções por Pestivirus/virologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Temperatura Corporal , Doença das Mucosas por Vírus da Diarreia Viral Bovina/imunologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/fisiopatologia , Brasil , Vírus da Diarreia Viral Bovina/isolamento & purificação , Modelos Animais de Doenças , Masculino , Infecções por Pestivirus/imunologia , Infecções por Pestivirus/veterinária , Fatores de Tempo , Carga Viral , Viremia/virologia , Eliminação de Partículas Virais
7.
J Dairy Sci ; 101(10): 9406-9418, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30031577

RESUMO

Determination of antimicrobial susceptibility (AMS) of Escherichia coli causing clinical mastitis (CM) according to the phylogenetic groups and its association with descriptors at the cow and herd level may help improve specific strategies for treatment and control of this pathogen in dairy herds. The aims of the present study were to (a) determine the frequency of phylogenetic groups of E. coli isolated from CM in dairy cows, and its association with cow-level descriptors (parity, lactation stage, CM severity, and affected quarter position), housing system, and season; and (b) determine and compare AMS among E. coli phylogenetic groups. A quadruplex PCR method was used to classify E. coli isolates into 1 of the 7 phylogenetic groups. Minimal inhibitory concentrations were determined for 10 antimicrobials, and survival analysis was performed to evaluate the AMS differences among E. coli phylogroups. Most E. coli isolates belonged to phylogroups A (52%) and B1 (38%). None of the cow- and herd-level descriptors were associated with the E. coli phylogenetic groups. Overall, E. coli isolates were mostly susceptible to ceftiofur (96.8%), sulfadimethoxine (75.5%), and cephalothin (74.5%). Based on the survival analysis, differences in AMS between phylogenetic groups of E. coli was observed only for cephalothin, in which strains of phylogroup A were inhibited at lower minimum inhibitory concentration than strains of phylogroup B1. Results of this study indicated low susceptibility of E. coli isolates identified from CM to most antimicrobials. In addition, differences in AMS can occur among E. coli phylogenetic groups, although they may be uncommon as they were limited to only one antimicrobial (i.e., cephalothin).


Assuntos
Antibacterianos/uso terapêutico , Infecções por Escherichia coli/veterinária , Escherichia coli/efeitos dos fármacos , Mastite Bovina/microbiologia , Testes de Sensibilidade Microbiana/veterinária , Animais , Bovinos , Escherichia coli/classificação , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/microbiologia , Feminino , Mastite Bovina/tratamento farmacológico , Filogenia , Gravidez
9.
J Dairy Sci ; 99(4): 2867-2874, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26830736

RESUMO

Bacteria adherence seems to be an essential first stage for the internalization of bacteria into the cytoplasm of the host cell, which is considered an important virulence strategy enabling bacteria to occupy a microenvironment separated from host defense mechanisms. Thus, this study aimed to explore the difference in the capacity of 4 bovine-associated staphylococci species or strains to adhere to and internalize into bovine mammary epithelial cells (MEC). Three different isolates of coagulase-negative staphylococci (CNS) were used: one strain of Staphylococcus fleurettii isolated from sawdust and considered an environmental opportunistic bacterium, and 2 dissimilar Staphylococcus chromogenes isolates, one cultured from a heifer's teat apex (Staph. chromogenes TA) and the other originating from a chronic intramammary infection (Staph. chromogenes IM). Also, one well-characterized strain of Staphylococcus aureus (Newbould 305) was used for comparison with a major mastitis pathogen. The CNS species and strains adhered to and internalized into MEC slower than did Staph. aureus. Still, we observed high variation in adhesion and internalization capacity among the different CNS, with Staph. chromogenes IM showing a greater ability to adhere to and internalize into MEC than the 2 CNS strains isolated from extramammary habitats. In conclusion, the 3 well-characterized bovine-associated CNS species and strains originating from distinct habitats showed clear differences in their capacity to adhere to and internalize into MEC. The observed differences might be related to their diversity in ecology and epidemiological behavior.


Assuntos
Células Epiteliais/microbiologia , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/fisiologia , Animais , Aderência Bacteriana , Bovinos , Contagem de Células , Células Cultivadas , Coagulase/metabolismo , Feminino , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/enzimologia , Staphylococcus/isolamento & purificação , Staphylococcus aureus/fisiologia
10.
J Dairy Sci ; 98(4): 2401-8, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25682144

RESUMO

An important question about intramammary infections that is still debated in the literature is the independence or interdependence of the quarters of dairy cows. The present study sought to explore milk neutrophil function and the milk lymphocyte profile of uninfected quarters from uninfected and infected (one infected quarter per cow) udders to evaluate interdependence of the quarters. Thus, 32 (8 cows) and 18 (6 cows) uninfected quarters from uninfected and infected udders were used, respectively. Using flow cytometry, we evaluated the percentage of milk neutrophils and their expression of adhesion molecules L-selectin (CD62L), ß2-integrin (CD11b), and an endothelial-selectin ligand (CD44); levels of intracellular reactive oxygen species (ROS); phagocytosis of Staphylococcus aureus by milk neutrophils; and neutrophil viability. Furthermore, we assessed the percentage of B-cell (CD21(+)) and T-lymphocyte subsets (CD3(+)/CD4(+)/CD8(-), CD3(+)/CD8(+)/CD4(-), CD3(+)/CD4(+)/CD25(-), CD3(+)/CD4(+)/CD25(+), and CD3(+)/CD4(-)/CD25(-)) using flow cytometry with monoclonal antibodies. The infected quarter did not affect somatic cell count or the percentage of neutrophils in the neighboring uninfected quarters. Furthermore, the infected quarter did not influence neutrophil viability, intracellular reactive oxygen species production, or phagocytosis of S. aureus by milk neutrophils. Conversely, the expression of adhesion molecules CD11b, CD62L, and CD44 by milk neutrophils differed between uninfected quarters from infected versus uninfected udders. The lymphocyte subsets did not differ between groups, except for a higher percentage of B cells in uninfected quarters from infected udders than in those from uninfected udders. Thus, our study strongly supports the hypothesis of interdependence of quarters based on the influence of infection on both the percentage of B cells and the expression of adhesion molecules by milk neutrophils in the neighboring uninfected quarters.


Assuntos
Citometria de Fluxo/veterinária , Glândulas Mamárias Animais/imunologia , Mastite Bovina/imunologia , Animais , Linfócitos B , Antígenos CD18 , Bovinos , Moléculas de Adesão Celular/análise , Feminino , Selectina L/imunologia , Contagem de Leucócitos/veterinária , Contagem de Linfócitos/veterinária , Subpopulações de Linfócitos/imunologia , Mastite Bovina/microbiologia , Mastite Bovina/fisiopatologia , Leite/citologia , Leite/imunologia , Neutrófilos/química , Neutrófilos/imunologia , Fagocitose/imunologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/imunologia , Linfócitos T
11.
Genet Mol Res ; 14(4): 14530-8, 2015 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-26600512

RESUMO

We conducted a phylogenetic analysis of 22 strains of bovine leukemia virus obtained by polymerase chain reaction to amplify a 582-base pair fragment of the transcriptional regulatory region 5' long terminal repeat (LTR). Twenty-two samples of proviral DNA from peripheral blood mononuclear cells containing bovine leukemia virus from naturally infected bovine from 4 distinct geographic regions in Brazil were investigated. The products obtained by polymerase chain reaction were subjected to direct sequencing and sequence alignment. Fragments of 422 nucleotides were obtained, located between positions -118 and +303 base pairs of the 5'LTR. These fragments corresponded to 80% of the LTR region and included 56% of sub-region U3, 100% of R, and 82.5% of U5. Phylogenetic analysis of these sequences showed a high conservation degree in the 5'LTR region, with 5 well defined groups. However, a hotspot occurrence in the R-U5 region was also observed, which contained 40% of all nucleotide variability observed.


Assuntos
Variação Genética , Vírus da Leucemia Bovina/genética , Filogenia , Sequências Repetidas Terminais/genética , Animais , Sequência de Bases , Brasil , Bovinos , DNA Viral/genética
12.
Genet Mol Res ; 13(1): 1717-23, 2014 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-24535910

RESUMO

This article reports the selection of bovine leukemia virus (BLV) variants after continuous passage in cell lines or experimental animals. Two wild BLV strains isolated from 2 naturally infected Holstein dairy cows in Brazil (cow codes: 485 and 141) were used for the experimental infection of 1 sheep and FLK cells, and 1 rabbit and CC81 cells. Viral DNA was isolated several months after infection, and env gene nucleotide and amino acid sequences of the "passaged" variants were compared against the 2 original infecting wild strains. The sequences of the original infecting wild strains were not recovered after their replication in the cell lines or experimental animals. These results indicate that genetic variation occurred after BLV replication in vivo and in vitro, with new variants being selected.


Assuntos
DNA Viral/genética , Genes env , Vírus da Leucemia Bovina/genética , Replicação Viral/genética , Animais , Sequência de Bases , Brasil , Bovinos , Divisão Celular , Linhagem Celular , Vírus da Leucemia Bovina/patogenicidade , Coelhos , Ovinos
13.
Genet Mol Res ; 13(2): 4607-16, 2014 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-25036510

RESUMO

This study aimed to develop and validate real-time PCR for the diagnosis of Mycobacterium bovis isolates. Two hundred and seventy-four M. bovis isolates and 156 M. tuberculosis isolates were tested. Both qPCRs amplified all of the 274 M. bovis samples, but none of the 156 M. tuberculosis samples. The qPCR for PE-PGRS 20 had 91% efficiency and a detection limit of 0.32 ng (sensitivity and specificity for qPCR "Mbovis.100" were 99.64 and 100%, respectively). The qPCR for RD4 had 100% efficiency, and a detection limit of 4 pg (diagnostic sensitivity and specificity were 100 and 100%. The qPCR tests were performed using 4 extraction sets, 3 qPCR kits, and with a range of equipment; yet, all combinations produced similar results in a diagnostic test, demonstrating the robustness of this method. The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.


Assuntos
Proteínas de Bactérias/genética , Mycobacterium bovis/genética , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Bovinos , DNA Bacteriano , Substâncias Intercalantes , Mycobacterium bovis/classificação , Mycobacterium bovis/isolamento & purificação , Mycobacterium tuberculosis/classificação , Mycobacterium tuberculosis/isolamento & purificação , Reprodutibilidade dos Testes , Tuberculose Bovina/microbiologia
14.
Data Brief ; 47: 108902, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36747980

RESUMO

A thorough characterization of base materials is the prerequisite for further research. In this paper, the characterization data of the reference materials (CEM I 42.5 R, limestone powder, calcined clay and a mixture of these three components) used in the second funding phase of the priority program 2005 of the German Research Foundation (DFG SPP 2005) are presented under the aspects of chemical and mineralogical composition as well as physical and chemical properties. The data were collected based on tests performed by up to eleven research groups involved in this cooperative program.

15.
Genet Mol Res ; 11(3): 2182-99, 2012 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-22653674

RESUMO

Equine infectious anemia caused by equine infectious anemia virus is an important disease due to its high severity and incidence in animals. We used a phage display library to isolate peptides that can be considered potential markers for equine infectious anemia diagnosis. We selected peptides using IgG purified from a pool comprised of 20 sera from animals naturally infected with equine infectious anemia virus. The diagnostic potential of these peptides was investigated by ELISA, Western blot and dot blot with purified IgG and serum samples. Based on the results, we chose a peptide mimetic for glycoprotein gp45 epitopes of equine infectious anemia virus, with potential for use as an antigen in indirect diagnostic assays. Synthesis of this peptide has possible applications for the development of new diagnostic tools for this disease.


Assuntos
Anemia Infecciosa Equina/sangue , Anemia Infecciosa Equina/diagnóstico , Cavalos/sangue , Cavalos/virologia , Peptídeos , Sequência de Aminoácidos , Animais , Western Blotting , Biologia Computacional , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/imunologia , Vírus da Anemia Infecciosa Equina/isolamento & purificação , Dados de Sequência Molecular , Biblioteca de Peptídeos , Peptídeos/química
16.
Genet Mol Res ; 11(3): 2051-9, 2012 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-22911589

RESUMO

Corynebacterium pseudotuberculosis, the infectious agent of caseous lymphadenitis (CLA), is responsible for substantial economic losses in goat and sheep production. Molecular characterization of C. pseudotuberculosis isolates by enterobacterial repetitive intergenic consensus (ERIC)-PCR has shown promising results in genotyping strains isolated from sheep with CLA. We evaluated the genetic diversity of C. pseudotuberculosis isolates collected from the Sertão region of the Pernambuco (PE) State, Brazil, and investigated the potential of ERIC-PCR as a tool for the molecular typing of strains of C. pseudotuberculosis isolated from goats. Thirty-two C. pseudotuberculosis strains isolated from goats in the municipalities of Floresta and Ibimirim, PE, C. pseudotuberculosis type strain ATCC 19410, the 1002 vaccine strain, and a field isolate of Rhodococcus equi were fingerprinted using the primers ERIC-1R and ERIC-2 and the primer pair ERIC- 1R+ERIC-2. Using 100% similarity as the cutoff, 8, 10, and 7 genotypes were obtained with ERIC-1-PCR, ERIC-2-PCR, and ERIC-1+2-PCR, respectively. The Hunter-Gaston discriminatory index calculated for the ERIC-1-PCR was 0.75. The index for the ERIC-2-PCR was 0.88, and the index for the ERIC-1+2-PCR was 0.79. Among goat isolates of C. pseudotuberculosis, three, two and four genotypes (found by ERIC-1-PCR, ERIC-2-PCR, and ERIC-1+2-PCR, respectively) had been previously described among sheep isolates from Minas Gerais State, Brazil. These results showed that ERIC-PCR has good discriminatory power and typeability, making it a useful tool for discrimination among C. pseudotuberculosis isolates from goats.


Assuntos
Sequência Consenso/genética , Corynebacterium pseudotuberculosis/genética , Corynebacterium pseudotuberculosis/isolamento & purificação , DNA Intergênico/genética , Cabras/microbiologia , Reação em Cadeia da Polimerase/métodos , Sequências Repetitivas de Ácido Nucleico/genética , Animais , Brasil , Análise por Conglomerados , Impressões Digitais de DNA , Enterobacteriaceae/genética
17.
Braz J Microbiol ; 43(4): 1632-40, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24031995

RESUMO

Suid herpesvirus 1 (SuHV-1) is the causative agent of pseudorabies (PR), a disease of great importance due to the huge losses it causes in the swine industry. The aim of this study was to determine a method for genotyping SuHV-1 based on partial sequences of the gene coding for glycoprotein C (gC) and to elucidate the possible reasons for the variability of this region. A total of 109 gCsequences collected from GenBank were divided into five major groups after reconstruction of a phylogenetic tree by Bayesian inference. The analysis showed that a portion of gC (approximately 671 bp) is under selective pressure at various points that coincide with regions of protein disorder. It was also possible to divide SuHV-1 into five genotypes that evolved under different selective pressures. These genotypes are not specific to countries or continents, perhaps due to multiple introduction events related to the importation of swine.

18.
Braz J Biol ; 84: e262851, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35674600

RESUMO

Nontuberculous mycobacteria infection is one of the most common chronic bacterial diseases in ornamental aquarium fish and appears to be directly related to stressful husbandry practices. Furthermore, it also represents zoonotic potential. Here we present the isolation and characterization of non-tuberculous mycobacteria from diseased freshwater angelfish (Pterophyllum scalare) in São Paulo, Brazil. Nine discarded breeding females with signs of disease were evaluated. The fish exhibited lethargy, loss of appetite, cachexia, skin ulcers, and exophthalmia. At necropsy, four fishes presented macroscopic granulomas in the spleen. Mycobacterium chelonae, M. fortuitum, M. gordonae, M. intracellulare and M. peregrinum were isolated and identified by hsp65 PCR restriction analysis. Histopathological analysis revealed microscopic lesions compatible with mycobacteriosis, and Mycobacterium bacillus were observed by Ziehl-Neelsen stain. Notably, all Mycobacterium species identified in this study have already been reported in human patients; therefore, diseased animals may be a source of infection for people who handle fish and aquariums.


Assuntos
Ciclídeos , Doenças dos Peixes , Infecções por Mycobacterium não Tuberculosas , Mycobacterium , Animais , Brasil , Água Doce , Humanos , Infecções por Mycobacterium não Tuberculosas/diagnóstico , Infecções por Mycobacterium não Tuberculosas/microbiologia , Infecções por Mycobacterium não Tuberculosas/veterinária
19.
Infect Genet Evol ; 103: 105322, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35753623

RESUMO

Cross-species transmission events and mixed infection of small ruminant lentiviruses (SRLVs) were studied in seven goats and two sheep from three small ruminant mixed flocks from Northeast and Southeast Brazil. Genetic and antigenic analyses with gag/env genes and ELISA multiepitope SU1/SU5 recombinant antigens were carried out, respectively. The genetic analysis of gag and env sequences showed high viral diversity in both species, MVV-like (subtype A1) and CAEV-like B1 in goats, and CAEV-like (subtype B1) in sheep, revealing SRLV interspecies transmission from sheep to goats and vice versa in Brazilian farms. Two Brazilian caprine lentiviruses were segregated in two new genetic clades based on gag analyses, which suggests a new classification into heterogenic genotype A. Furthermore, goat isolates were grouped into subtype A1 and B1 clusters. Cross-reactive antibodies were detected in goats using ELISA with a recombinant antigen carrying SU1 and SU5 immunodominant epitopes; the results showed anti-CAEV and MVV antibodies in goats and anti-CAEV antibodies in sheep. This result can be associated with the high divergence in the V4 region due to SRLV variability. All results confirm cross-species infection of SRLV in Brazilian mixed herds.


Assuntos
Doenças das Cabras , Infecções por Lentivirus , Doenças dos Ovinos , Animais , Brasil/epidemiologia , Cabras , Lentivirus/genética , Infecções por Lentivirus/veterinária , Filogenia , Ruminantes , Ovinos
20.
Chirurgie (Heidelb) ; 93(10): 976-982, 2022 Oct.
Artigo em Alemão | MEDLINE | ID: mdl-35925137

RESUMO

BACKGROUND: The COVID-19 pandemic has changed medical teaching worldwide. Digital teaching and examinations are successful for cognitive learning objectives, whereas practical skills had to be carried out predominantly in person under strict hygiene standards. AIM: This study presents the opportunities and challenges of using a presence objective structured clinical examination (OSCE) at a distance with digital support. METHOD: Following surgical practical teaching an OSCE was conducted in presence, where students demonstrated practical skills in one room while the examiners were connected via videoconference from another room. Students were surveyed about the OSCE and sustained learning via a standardized online questionnaire after completion of the surgical teaching. Additionally, examiners were surveyed on their experiences. RESULTS: In the online survey 40 students participated (25% of n = 157 students contacted) and 36 completed questionnaires were analyzed. Overall, the implementation of an OSCE even under pandemic conditions was perceived as very positive by the students (92% of students n = 33). In particular, the acquisition of practical skills was rated as very high. For 78% (n = 28) of the students, the acquisition of competencies through the practical examination was particularly sustainable. The vast majority of students and examiners felt safe regarding infection control because of the hygiene concept (92%, n = 33). Overall, 80 students achieved grade 1 (51%), 71 students grade 2 (45.2%) and 6 students grade 3 (3.8%) (grade 1 = very good, grade 6 = very bad). CONCLUSION: Practical examinations are essential for checking practical learning objectives and can be implemented at a distance with a well-developed hygiene concept and digital support.


Assuntos
COVID-19 , Educação Médica , COVID-19/epidemiologia , Humanos , Pandemias , Exame Físico
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