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1.
Burns ; 50(3): 685-690, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38042627

RESUMO

OBJECTIVE: Commercially available bioethanol-fueled fireplaces are a potential source of burns and are commonly used for home use. The present study aimed to evaluate the quality of life following burn injuries that were caused by bioethanol-related accidents. METHODS: Burned patients who were admitted to our burn unit with burn injury due to bio-ethanol fueled fire places between January 2010 and December 2021 were contacted to ask for their willingness to participate in this study. They were asked to answer questions regarding the circumstances of the accident and three questionnaires to capture burn specific and general health related quality of life (Burn Specific Health Scale-Brief (BSHS-B), Short-Form Health Survey 36 (SF-36)) and general information about the accident. Patients were matched and compared to a group of patients suffering comparable burns from other burn mechanisms, which were also admitted to our burn unit at the same time. RESULTS: Of 35 patients that met the inclusion criteria, 19 answered the questionnaire and were compared to 38 patients with other burn mechanisms. There were no statistical differences regarding age (bioethanol: 37.4 ± 14.7 years vs. control: 36.2 ± 14.3 years, p = 0.777), TBSA (9.9 ± 6.8% vs. 8.9 ± 10.4, p = 0.715), and sex (42.1% females vs. 36.8% females, p = 0.882). Most patients in the bioethanol-group reported that they did not follow the manual instructions (68.4%) and that the accident happened during the refilling process (52.6%). There was no significant difference in any subscale of the BSHS-B or the SF-36. DISCUSSION: Burns related to bioethanol-fueled fireplaces are rare compared to other typical burn mechanisms. However, as they are used for personal pleasure and interior design, psychological impairment following burn may be even more critical. Detailed education on the use of these fireplaces needs to take place in order to reduce the risk of accidents.


Assuntos
Queimaduras , Qualidade de Vida , Feminino , Humanos , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Masculino , Qualidade de Vida/psicologia , Queimaduras/epidemiologia , Produtos Domésticos , Etanol/efeitos adversos , Ansiedade , Inquéritos e Questionários
2.
Unfallchirurg ; 113(11): 915-22, 2010 Nov.
Artigo em Alemão | MEDLINE | ID: mdl-20703444

RESUMO

Severe burn injuries are traumatic events and can have serious impact on all areas of life frequently causing high emotional distress. In a multicentre study resources and emotional distress of patients with serious burn injuries were assessed during the first hospitalization and at 6 and 12 months follow-up. Patients with severe burn injuries after accidents in a private environment (NBG patients) and patients after occupational accidents covered by the German Social Accident Insurance (BG patients) were compared. All patients reported marked emotional impairment, particularly during the hospitalization. At follow-up a reduction of emotional distress was detected. Nearly half of the patients received a diagnosis of one or more mental disorders according to DSM-IV criteria. When treating patients with burns, special attention should be given to their mental health. They should be offered psychological support to cope with the aftermath of the accident, especially after discharge from hospital when returning to their normal surroundings.


Assuntos
Queimaduras/epidemiologia , Queimaduras/psicologia , Previdência Social/estatística & dados numéricos , Estresse Psicológico/epidemiologia , Estresse Psicológico/psicologia , Adolescente , Adulto , Comorbidade , Feminino , Alemanha/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Medição de Risco , Fatores de Risco , Adulto Jovem
3.
Int J Food Microbiol ; 135(3): 199-202, 2009 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-19723600

RESUMO

Between October 2007 and March 2008, 153 wild boars shot in the Canton of Geneva in Switzerland were sampled. Fifty-one percent of the animals were males and 49% were females. The age of most (81%) animals varied between 6months and 2years. Prevalence of enteropathogenic Yersinia in tonsils and faeces was studied using culture and PCR methods and in tissue fluid of tonsils using an ELISA system. Prevalence of anti-Yersinia antibodies in tissue fluid was 65%. Detection rate of enteropathogenic Yersinia in tonsils of 153 wild boars by real-time PCR was 44%. Ail-positive Yersiniaenterocolitica and inv-positive Yersiniapseudotuberculosis were detected in 35 and 20% of the animals, respectively. Both species were detected in 10% of the animals. Isolation rate of enteropathogenic Yersinia was low; ail-positive Y. enterocolitica and inv-positive Y. pseudotuberculosis were found in 9 and 3% of the animals, respectively. Prevalence was shown to be significantly higher in tonsils than in faeces. Furthermore, females were more commonly positive than males. This study shows that the prevalence of enteropathogenic Yersinia is high and both enteropathogenic Y. enterocolitica and Y. pseudotuberculosis are common findings in tonsils of wild boars in Switzerland.


Assuntos
Fezes/microbiologia , Tonsila Palatina/microbiologia , Sus scrofa/microbiologia , Yersiniose/veterinária , Yersinia enterocolitica/isolamento & purificação , Yersinia pseudotuberculosis/isolamento & purificação , Animais , Anticorpos/metabolismo , DNA Bacteriano , Ensaio de Imunoadsorção Enzimática , Feminino , Masculino , Tonsila Palatina/imunologia , Reação em Cadeia da Polimerase , Prevalência , Suíça/epidemiologia , Yersiniose/epidemiologia , Yersinia enterocolitica/genética , Yersinia enterocolitica/imunologia , Yersinia pseudotuberculosis/genética , Yersinia pseudotuberculosis/imunologia
4.
Neuroscience ; 157(2): 385-95, 2008 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-18832015

RESUMO

In this study we evaluated the effects of the novel, potent non-competitive metabotropic glutamate receptor (mGluR) 1 antagonist (3aS,6aS)-6a-naphthalen-2-ylmethyl-5-methyliden-hexahydro-cyclopental[c]furan-1-on (BAY 36-7620) on different types of synaptic plasticity in the hippocampal cornu ammonis (CA) 1-region and on hippocampus-dependent spatial learning. After having confirmed the presence of mGluR1 in the hippocampal CA1 region of our rat strain by confocal microscopy, we tested the effects of BAY 36-7620 on: 1) long-term potentiation (LTP) induced by weak and strong stimulation; 2) 3,5-dihydroxyphenylglycine (DHPG, 30 microM)-induced depression of synaptic transmission; and 3) learning of the hidden platform version of the water maze by mice. BAY 36-7620 (10 microM) amplified LTP but, like the mGluR1 antagonists 7-hydroxyiminocyclopropan[b]chromen-1a-carboxylic acid ethyl ester (CPCCOEt, 10 microM) and 4-carboxyphenylglycine (4-CPG, 50 microM), diminished LTP at 1 microM. The mGluR5 antagonist 6-methyl-2-(phenylethynyl)-pyridine (MPEP, 10 microM) had no effect. BAY 36-7620 (10 microM) did not affect strong LTP. Thus, mGlu 1, but not mGlu 5, receptors modulate LTP elicited by weak stimulation in vitro. DHPG-induced depression of synaptic transmission was only marginally affected by BAY 36-7620 (1 microM) or 4-CPG (100 microM). In a mouse water maze study, BAY 36-7620 (10 mg/kg, i.v.) increased the escape latency and impaired water escape task acquisition during the first 4 days. Drug- and vehicle-treated groups showed comparable performance at day 5. Our data support a role for mGluR1 in LTP and in the acquisition of spatial memory.


Assuntos
Hipocampo/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacos , Naftalenos/farmacologia , Plasticidade Neuronal/efeitos dos fármacos , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Transmissão Sináptica/efeitos dos fármacos , Análise de Variância , Animais , Estimulação Elétrica , Antagonistas de Aminoácidos Excitatórios/farmacologia , Hipocampo/citologia , Masculino , Camundongos , Camundongos Knockout , Técnicas de Patch-Clamp , Ratos , Receptores de Glutamato Metabotrópico/deficiência
5.
Int J Food Microbiol ; 95(1): 89-94, 2004 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-15240078

RESUMO

The distribution of Yersinia spp. in butcher shops in the Munich area was studied. The isolates recovered were then characterised with pulsed-field gel electrophoresis (PFGE) to identify possible contamination routes. A total of 298 samples were collected from eight small butcher shops between June and August in 2001. Of these, 113 were surface samples from carcasses, offal and raw pork products, and 185 were environmental surface samples from tools, equipment and processing areas. The samples were studied with direct plating, overnight enrichment in nonselective broth and selective enrichment in two different enrichment broths. The Yersinia isolates recovered were characterised with PFGE using NotI, ApaI and XhoI DNA restriction enzymes. Yersinia was recovered from all eight butcher shops, and pathogenic Yersinia enterocolitica 4/O:3 was present in six shops. The occurrence of this pathogen on raw pork products varied from 8% to 25%. Pathogenic Y. enterocolitica 4/O:3 was isolated from two environmental samples: a worktable and a chain glove. Most (18/24) of the Yersinia-positive samples were found already after direct plating. Forty-nine Yersinia isolates from 24 samples were studied with PFGE. Twelve genotypes (I-XII) were obtained among Y. enterocolitica 4/O:3 when 33 isolates from 16 samples were characterised with NotI, ApaI and XhoI enzymes. The genotypes of Y. enterocolitica 4/O:3 strains differed among butcher shops. In most (5/6) shops, more than one genotype was found, indicating different contamination sources. In conclusion, raw pork products from butcher shops are frequently contaminated with different genotypes of pathogenic Y. enterocolitica 4/O:3, thus serving as an important transmission vehicle from butcher shops to humans.


Assuntos
Qualidade de Produtos para o Consumidor , Contaminação de Equipamentos , Carne/microbiologia , Yersinia enterocolitica/genética , Animais , Eletroforese em Gel de Campo Pulsado , Contaminação de Alimentos , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Genótipo , Alemanha , Humanos , Produtos da Carne/microbiologia , Suínos , Yersiniose/transmissão , Yersinia enterocolitica/classificação , Yersinia enterocolitica/isolamento & purificação
6.
Syst Appl Microbiol ; 26(1): 132-7, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12747421

RESUMO

The distribution of different genotypes of Y. enterocolitica 4:O3 strains recovered from pig tonsils in Southern Germany and Finland in 1999-2000 was investigated. A total of 96 and 207 Y. enterococolitica 4:O3 isolates recovered from 47 and 66 tonsils of finishing pigs in Germany and Finland, respectively, were characterised with PFGE using NotI enzyme. In all, 39 different NotI profiles were obtained, only one of which, NB1, was found in both Germany and Finland. All strains were further characterised with ApaI and XhoI enzymes. When the 54 German and 74 Finnish strains were characterised with all three enzymes, 51 genotypes were obtained. The 23 genotypes found in German strains differed from the 28 found in Finnish strains. These results indicate that Y. enterocolitica 4:O3 genotypes have a differential geographical distribution and thus can be used in epidemiological studies.


Assuntos
Microbiologia de Alimentos , Tonsila Palatina/microbiologia , Suínos/microbiologia , Yersinia enterocolitica/isolamento & purificação , Matadouros , Animais , Desoxirribonucleases de Sítio Específico do Tipo II/genética , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Eletroforese em Gel de Campo Pulsado/métodos , Finlândia , Genótipo , Alemanha , Humanos , Yersinia enterocolitica/enzimologia , Yersinia enterocolitica/genética
7.
Int J Food Microbiol ; 82(1): 81-6, 2003 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-12505462

RESUMO

In order to detect phosphatase activity a total of 137 isolates from 12 Clostridium (C.) species were examined via fluorescence on SCA-agar with methylumbelliferyl phosphate (SCA-MUP), via APIZYM and RAPID ID 32 A as well as using a phosphatase reagent containing 1-naphthyl phosphate. Fluorescence on SCA-MUP showed the presence of acid or alkaline phosphatase in almost all isolates examined. Likewise, acid or alkaline phosphatase could be detected via APIZYM and RAPID ID 32 A in most strains and species. Opposed to this, the majority of the Clostridium bifermentans isolates showed a positive reaction exclusively on SCA-MUP. On the other hand, the phosphatase reagent for the detection of acid phosphatase lead to unambiguously positive results only when examining Clostridium perfringens isolates. Therefore, the SCA-MUP-agar, in contrast to the phosphatase reagent, was proven to be unsuitable for the identification of C. perfringens via detection of acid phosphatase. Using the phosphatase reagent the activity of this enzyme was detected in 95.1% of the C. perfringens isolates included in the study. In addition, the phosphatase reagent showed identical reactions after a 24 h incubation at 37 degrees C and when used on cultures incubated for 6 h at 44 degrees C in the case of 98.5% of the C. perfringens isolates.


Assuntos
Clostridium perfringens/enzimologia , Microbiologia de Alimentos , Monoéster Fosfórico Hidrolases/isolamento & purificação , Fluorescência , Monoéster Fosfórico Hidrolases/metabolismo , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade
8.
Lett Appl Microbiol ; 34(4): 238-43, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11940151

RESUMO

AIMS: Clostridium (Cl.) perfringens is a common cause of food poisoning outbreaks. Ribosomal DNA analysis (ribotyping), a method which analyses restriction fragment length polymorphisms in the chromosomal genes that encode rRNA, has been shown to be useful for microbial species identification and subtyping. METHODS AND RESULTS: The current study has used ribotyping to examine 111 Cl. perfringens isolates from industrially produced ground meat in order to collect a basis for a contamination survey. Among the 111 isolates 107 distinctly different ribopatterns were detected. In only four cases two Cl. perfringens isolates showed an identical ribopattern. The isolates gave identical ribotype patterns in three different runs, carried out 3-4 months apart from each other. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The discriminatory index for EcoRI ribotyping of the Cl. perfringens isolates was 0 x 99. Results showed that ribotyping is suitable for subtyping Cl. perfringens isolates from raw meat. Ribotyping appeared to be a useful tool for profound epidemiologic studies of Cl. perfringens-contamination in food production and processing.


Assuntos
Clostridium perfringens/isolamento & purificação , Produtos da Carne/microbiologia , Animais , Bovinos , Clostridium perfringens/genética , Eletroforese em Gel de Poliacrilamida , Indústria Alimentícia , Microbiologia de Alimentos , Genes Bacterianos , Humanos , Filogenia , Ribotipagem/métodos
9.
J Food Prot ; 64(9): 1435-8, 2001 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11563524

RESUMO

Contamination of minced meat with Salmonella is still considered a major problem in food hygiene. Therefore, in this study the Salmonella incidence in minced meat produced in a European Union-approved slaughtering and cutting plant was investigated in detail. Throughout 21 months, 297 pool samples (1,485 individual samples) of mixed minced meat (beef and pork) were examined according to Council Directive 94/65/EC and to ISO 6579. Salmonellae were detected in 47 (15.8%) of the pool samples. After separation of the positive pools, 93 individual samples were determined to be Salmonella positive, representing 6.3% of the total 1,485 samples. Serotyping resulted in most isolates (69.6%) being identified as Salmonella Typhimurium. It was further shown that the incidence of Salmonella isolations varied during the year and that the isolation rate was higher on some days of the week compared with others.


Assuntos
Produtos da Carne/microbiologia , Salmonella/isolamento & purificação , Animais , Bovinos , União Europeia , Contaminação de Alimentos , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Incidência , Sorotipagem , Suínos
10.
Electrophoresis ; 22(8): 1585-9, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11386674

RESUMO

Genotyping of bacterial strains via pulsed-field gel electrophoresis has to be considered an important tool for epidemiological investigations in food hygiene as well as in other areas. Yet, a major disadvantage of this method is its long duration. Therefore, rapid procedures for DNA isolation and restriction are being sought. One such protocol was modified and further shortened to two days. This short protocol was used for macrorestriction analysis of 34 strains of 25 different Clostridium species. Parallel analyses were performed using a conventional 5-day protocol in order to compare the long and the short method by running the DNA samples obtained via both protocols on the same gel. In the case of nine strains, none of the two methods yielded satisfactory results, whereas for three strains the long protocol proved to be preferable to the short one. Comparable results were obtained using both methods in the case of 22 strains belonging to 17 different Clostridium species.


Assuntos
Clostridium/genética , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado/métodos , Clostridium/classificação
11.
Mol Pharmacol ; 59(5): 965-73, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11306677

RESUMO

L-Glutamate (Glu) activates at least eight different G protein-coupled receptors known as metabotropic glutamate (mGlu) receptors, which mostly act as regulators of synaptic transmission. These receptors consist of two domains: an extracellular domain in which agonists bind and a transmembrane heptahelix region involved in G protein activation. Although new mGlu receptor agonists and antagonists have been described, few are selective for a single mGlu subtype. Here, we have examined the effects of a novel compound, BAY36-7620 [(3aS,6aS)- 6a-Naphtalen-2-ylmethyl-5-methyliden-hexahydro-cyclopental[c]furan-1-on], on mGlu receptors (mGlu1-8), transiently expressed in human embryonic kidney 293 cells. BAY36-7620 is a potent (IC(50) = 0.16 microM) and selective antagonist at mGlu1 receptors and inhibits >60% of mGlu1a receptor constitutive activity (IC(50) = 0.38 microM). BAY36-7620 is therefore the first described mGlu1 receptor inverse agonist. To address the mechanism of action of BAY36-7620, Glu dose-response curves were performed in the presence of increasing concentrations of BAY36-7620. The results show that BAY36-7620 largely decreases the maximal effect of Glu. Moreover, BAY36-7620 did not displace the [(3)H]quisqualate binding from the Glu-binding pocket, further indicating that BAY36-7620 is a noncompetitive mGlu1 antagonist. Studies of chimeric receptors containing regions of mGlu1 and regions of DmGluA, mGlu2, or mGlu5, revealed that the transmembrane region of mGlu1 is necessary for activity of BAY36-7620. Transmembrane helices 4 to 7 are shown to play a critical role in the selectivity of BAY36-7620. This specific site of action of BAY36-7620 differs from that of competitive antagonists and indicates that the transmembrane region plays a pivotal role in the agonist-independent activity of this receptor. BAY36-7620 will be useful to further delineate the functional importance of the mGlu1 receptor, including its putative agonist-independent activity.


Assuntos
Naftalenos/farmacologia , Receptores de Glutamato Metabotrópico/agonistas , Animais , Células Cultivadas , Humanos , Fosfatos de Inositol/metabolismo , Proteínas de Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Ratos , Receptores de Glutamato Metabotrópico/metabolismo , Transfecção
12.
Syst Appl Microbiol ; 24(3): 457-63, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11822684

RESUMO

Prevalence and contamination routes of pathogenic Yersinia enterocolitica were studied in Southern Germany. Tonsil and faeces samples of 50 fattening pigs, 140 offal samples and 120 minced meat samples were examined. Pig and offal samples were collected from a slaughterhouse approved by the European Union, and minced meat samples from two large meat factories. Yersinia enterocolitica was isolated using direct plating, overnight enrichment and selective enrichment in MRB and ITC broth. The isolates were bio- and serotyped, and pathogenicity was studied using two plasmid-encoded virulence markers: calcium dependence and Congo red absorption. The genotypes were studied with pulsed-field gel electrophoresis using NotI enzyme. Prevalence of pathogenic Y. enterocolitica 4:O3 was 60% and 10% in tonsils and faeces of fattening pigs, respectively. Besides tonsils, prevalence of pathogenic Y. enterocolitica 4:O3 was also high in other pluck set samples, including tongues, lungs, hearts, diaphragms and livers. However, the highest isolation rate was obtained from the tonsils. Kidneys, which were not attached to the pluck set and did not hang together with tonsils on the rack, had the lowest isolation rate. Yersinia enterocolitica 4:O3 was isolated from 12% of minced meat samples. A total of 25 NotI profiles were obtained from porcine samples. The most common genotype, NBI, found in tonsils was also the most common type recovered from offal and minced meat samples. The high contamination rate of tonsils, and the indistinguishable NotI profiles obtained from tonsils and offal indicate that the tonsils contaminate offal when they are removed and hung on the rack together. When the head, with the tonsils and tongue, is not removed prior to evisceration and is not handled and inspected separately, it is difficult to control the spread of Y. enterocolitica 4:O3 from tonsils to the carcass, and subsequently, to meat.


Assuntos
Matadouros , Suínos/microbiologia , Yersinia enterocolitica/isolamento & purificação , Animais , Alemanha , Prevalência
13.
J Food Prot ; 62(12): 1488-96, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10606159

RESUMO

Impedance microbiology is a rapid method that enables qualitative and quantitative tracing of microorganisms by measuring the change in the electrical conductivity. With direct impedance technology, the change in the conductivity of a liquid culture medium serves as a measuring parameter, whereas with indirect impediometry, the change in the electrical conductivity of a reaction solution, which occurs through the absorption of gases from the inoculated bacterial culture, is measured. Most investigations concerning the applicability of impediometry in food microbiology deal with the impedimetric detection or enumeration of Enterobacteriaceae, especially the detection of Salmonella. However, impediometry has been applied to other bacterial groups or species as well. Furthermore, a great number of published findings concern the impedimetric determination of the total bacterial count. The successful application of this fast method on further areas of food hygiene, such as tracing antibiotics and testing additives for their antimicrobiological effect, has also been described. In general the use of impediometry for the application areas stated has been judged positively. However, the time and expense required by the user to optimize the method, the deficits when testing slightly contaminated sample material or determining the bacterial count in those cases in which the microorganisms are sublethally damaged, and the necessity of performing individual calibration for each food category limit the applicability of impediometry.


Assuntos
Bactérias/isolamento & purificação , Técnicas Bacteriológicas , Condutometria , Impedância Elétrica , Microbiologia de Alimentos , Condutometria/instrumentação , Condutometria/métodos
14.
FEMS Immunol Med Microbiol ; 24(3): 281-6, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10397312

RESUMO

Clostridium perfringens continues to be a common cause of food-borne disease. It produces an enterotoxin (CPE) which is released upon lysis of the vegetative cell during sporulation in the intestinal tract. Catering premises with insufficient cooling and reheating devices often seem to be the cause of outbreaks of C. perfringens food poisoning. Typing of C. perfringens is of great importance for investigating sources of food poisoning cases and for studying the epidemiology of this microorganism. This report describes the examination of 155 C. perfringens isolates by molecular methods. Isolates were taken from 10 food poisoning outbreaks and cases (n = 34, food and fecal isolates) and from meat and fish pastes (n = 121). Isolates were characterized by plasmid profiling, ribotyping, and/or macrorestriction analysis by pulsed-field gel electrophoresis (PFGE). Results show that all three methods are suitable for classifying C. perfringens isolates below the species level. Ribopatterns and PFGE patterns can be interpreted more easily than plasmid profiling results and can be recommended for contamination studies and epidemiologic investigation of food poisonings associated with C. perfringens.


Assuntos
Infecções por Clostridium/microbiologia , Clostridium perfringens/genética , Surtos de Doenças , Animais , Bovinos , Galinhas , Infecções por Clostridium/epidemiologia , Clostridium perfringens/isolamento & purificação , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Carne/microbiologia , Plasmídeos , Coelhos
15.
FEMS Immunol Med Microbiol ; 24(3): 287-92, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10397313

RESUMO

While pulsed field gel electrophoresis has become an important tool for genotyping of bacteria, one of its drawbacks is that standard methods are rather time-consuming. In order to overcome this problem, shortened procedures for DNA preparation have been developed for some bacterial species. The aim of this study was to examine if a short procedure used for pulsed field gel electrophoresis of Clostridium botulinum could be applied to other Clostridia species. For this, the protocol was modified and used to prepare the DNA of 34 strains of 25 different Clostridia species. In contrast to a standard procedure, which takes at least 5 days from DNA extraction to completion of the electrophoresis, this protocol yielded results within 2 days. In order to directly compare the results of the short protocol with those of the standard, long procedure, parallel DNA preparations were performed using both methods and the two DNA samples thus obtained per strain were then run on the same gel. Briefly, the procedure was as follows. After embedding the bacterial cells in agarose, the agarose blocks were incubated for 1 h in lysis solution containing lysozyme, mutanolysin, lysostaphin and RNase. This was followed by a 1-h proteinase K treatment. Then, slices were cut from the agarose blocks and washed for 15 min in TE buffer, these washes were repeated four times with fresh TE. After a 2-h restriction with SmaI, electrophoresis was carried out overnight.


Assuntos
Clostridium/genética , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado/métodos
16.
Int J Food Microbiol ; 52(3): 169-80, 1999 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-10733248

RESUMO

The identification of Clostridium species using conventional biochemical reactions or commercially available miniaturized ready-to-use test kits often yields uncertain results. One of these test kits, the RAPID ID 32 A identification system for anaerobes (bioMérieux, Marcy-l'Etoile, France), is based on the evaluation of the action of preformed bacterial enzymes and allows classification to the species level within 4 h. This study intended to assess the suitability and reliability of this system for the rapid identification of Clostridium species relevant to food hygiene. For this purpose, 122 test strains of 18 different Clostridium species were examined via RAPID ID 32 A. Of these, 110 (90.2%) were correctly identified to the species level within 4 h. In addition, six strains were successfully classified after examination with supplementary biochemical reactions suggested by the manufacturer, which required an overall examination time of 72 h. The identity of five Clostridium strains could not be determined and in one case the test kit yielded a wrong species classification. Altogether, a correct identification was possible for 116 (95.1%) of the isolates, the total error rate was 4.9%. Using the RAPID ID 32 A system a great number of Clostridium species which are relevant to food hygiene and are often difficult to identify can be correctly and reliably classified within 4 h. Nevertheless, the test system has to be regarded as not yet completely satisfactory, e.g., because C. tyrobutyricum isolates were only insufficiently identified. Further improvements of the system are desirable in order to make it universally applicable in food microbiology.


Assuntos
Clostridium/isolamento & purificação , Microbiologia de Alimentos , Kit de Reagentes para Diagnóstico/normas , Clostridium/classificação , Carne/microbiologia
17.
J Food Prot ; 61(2): 240-3, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9708289

RESUMO

The prevalence of the enterotoxin gene in a well-characterized collection of 71 Clostridium perfringens strains from 36 separate food-poisoning cases or outbreaks was analyzed with the polymerase chain reaction (PCR). The clonality of 39 strains originating from 14 outbreaks where at least two isolates were available was studied with pulsed-field gel electrophoresis (PFGE) using SmaI and ApaI restriction endonucleases. The cpe gene PCR assay was found to correlate well with Clostridium perfringens enterotoxin (CPE) production in vitro with reverse passive latex agglutination. Of the C. perfringens food and clinical food-poisoning isolates 24 (86%) and 38 (88%) were cpe-positive, respectively. Different PFGE patterns indicated that multiple cpe-positive clones are frequently present within one outbreak. The existence of cpe-positive and negative isolates with identical or nearly identical PFGE patterns in a single outbreak suggests that the cpe gene may be in a movable genetic element.


Assuntos
Infecções por Clostridium/epidemiologia , Clostridium perfringens/genética , Surtos de Doenças , Enterotoxinas/genética , Doenças Transmitidas por Alimentos/epidemiologia , Animais , Clonagem Molecular , Desoxirribonucleases de Sítio Específico do Tipo II , Enterotoxinas/química , Fezes/microbiologia , Alemanha , Humanos , Produtos da Carne/microbiologia , Produtos Avícolas/microbiologia
18.
Zentralbl Veterinarmed B ; 45(10): 595-602, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9916550

RESUMO

Bacteriocin typing, plasmid profiling and ribotyping were used to type 34 food and patient Clostridium perfringens isolates from 10 food poisoning cases, respectively, outbreaks. In nine cases/outbreaks bacteriocin patterns showed identical main groups. Subgroups differed within all cases/outbreaks. Plasmid profiles were identical for all isolates within each of three outbreaks. In eight food poisoning cases and outbreaks, all the ribotypes of each food and stool isolate were found to be identical. All three typing methods give valuable results for the characterization of C. perfringens beyond the species level. Bacteriocin typing represents a suitable addition to plasmid typing, particularly since the results do not show any correlation between losses of plasmids and changes in bacteriocin sensitivity patterns. Ribotyping was found to be a suitable tool to determine the genetic relationship of C. perfringens isolates in the context of food-borne poisoning.


Assuntos
Infecções por Clostridium/microbiologia , Clostridium perfringens/classificação , Doenças Transmitidas por Alimentos/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Bacteriocinas/análise , DNA Bacteriano/análise , Fezes/microbiologia , Humanos , Plasmídeos/classificação
19.
Appl Environ Microbiol ; 63(10): 3992-4, 1997 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9327563

RESUMO

Ribotyping was used to characterize 34 Clostridium perfringens strains isolated from 10 food poisoning cases and outbreaks over a 7-year period. Twelve different ribopatterns were generated by EcoRI digestion. In eight food poisoning cases and outbreaks, all of the ribotypes of each food and stool isolate were found to be identical. Two C. perfringens isolates showed unique patterns. Ribotyping was found to be a useful tool for determining the genetic relationship of C. perfringens isolates in the context of foodborne poisoning cases.


Assuntos
Infecções por Clostridium/microbiologia , Clostridium perfringens/classificação , Clostridium perfringens/genética , Doenças Transmitidas por Alimentos/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Infecções por Clostridium/epidemiologia , Clostridium perfringens/isolamento & purificação , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Desoxirribonuclease EcoRI , Surtos de Doenças , Fezes/microbiologia , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/epidemiologia , Alemanha/epidemiologia , Humanos
20.
Zentralbl Veterinarmed B ; 44(4): 221-33, 1997 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9230673

RESUMO

A procedure is given for differentiation of motile Aeromonas spp. Based on nine to 12 biochemical properties it allows an assignment of isolated Aeromonas strains first to phenotypes and, in the second step, to hybridization groups (HGs) 1-13 as well as to the genotypically defined species Aer. allosaccharophila and Aer. encheleia. The computer aided classification is carried out according to the principles of numerical taxonomy. Using this differentiation scheme 23 Aeromonas strains, which were isolated from food, and two reference strains were speciated. A total of 16 strains were assigned to Aer. hydrophila, six strains to Aer. caviae and one strain to Aer. sobria. Of the 16 Aer. hydrophile strains, 11 were attached to HG 1, one to HG 2 and four could not be classified clearly as HG 3 or HG 1. All six Aer. caviae isolated were assigned to HG 5 and the Aer. sobria strain was ranked among HG 8. The reference strains were assigned to the same genotypes by this method than by DNA-DNA hybridization assays.


Assuntos
Aeromonas/classificação , Galinhas/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Doenças das Aves Domésticas/diagnóstico , Aeromonas/genética , Aeromonas/metabolismo , Animais , DNA Bacteriano/análise , DNA Bacteriano/genética , Diagnóstico por Computador/métodos , Diagnóstico por Computador/veterinária , Microbiologia de Alimentos , Genótipo , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/microbiologia , Técnicas Microbiológicas/veterinária , Hibridização de Ácido Nucleico , Fenótipo , Doenças das Aves Domésticas/microbiologia
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