RESUMO
Culture conditions were optimized for toxin production of Clostridium botulinum type G, the last toxigenic serotype described. Six factorial experiments were performed to assess the effect of medium composition (nutrients, metal ions, sterile soil, pH), incubation conditions (time, temperature and anaerobiosis) and associated microorganisms (Bacillus subtilis, Lactobacillus plantarum) on the toxinogenesis of C. botulinum type G. A significant (4 to 10 fold) improvement of toxin production was obtained by using an optimized medium (3% proteose-peptone, 0.5% trypticase, 1.1% glucose, 0.5% yeast extract, adjusted to pH 8.0) and incubation of the culture for 12 days at 26 degrees C in a nitrogen atmosphere.
Assuntos
Toxinas Botulínicas/biossíntese , Clostridium botulinum/metabolismo , Anaerobiose , Meios de Cultura , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
In order to detect subclinical mastitis by means of California Mastitis Test and recounting of somatic cells, 163 cows from the dairies of San Luis city, Argentina, were examined. Seventy six individuals (46.6%) exhibited an inflammatory response ranging > or = 2+ grade and a cellular recounting value of > or = 5 x 10(5), data compatible with those of subclinical mastitis. Staphylococcus aureus was isolated from 39 (51.3%) cultures as estimated by the sum of the two last values listed in Table 1. Organisms were isolated by plating on brain heart infusion agar with 5% of sheep blood and on Baird-Parker media. One hundred and three S. aureus isolates recovered from 51 of 63 cows were characterized by coagulase activity by the tube method using human and bovine plasma; clumping factor; glucose and mannitol fermentation; thermonuclease (TNase), pigment, gelatinase, fibrinolysin, acetoin, hemolysin production; egg yolk, tellurite and catalase reaction and crystal violet types. All isolates were susceptible to cephalothin, clindamycin, methicillin, gentamycin and vancomycin; 94.1% were susceptible to chloramphenicol and 53.8% to G penicillin. Sixty three isolates (61.1%) were classified according to Hájek and Marsálek scheme as biotype C (bovine and ovine ecovar), 33 isolates (32.0%) were classified as biotype B (swine and poultry ecovar); 1 isolated (0.9%) as intermediate between B and D; 5 isolates (4.8%) as biotype A (human ecovar) and 1 isolated (0.9%) as biotype D (ecovar silvestres spp) (Table 2). Production of enterotoxins A to E and toxic shock syndrome toxin-1 (TSST-1) was determined by the optimal susceptibility plate method on 27 isolates (26.2%) which were coagulase 3+ to 4+ and TNase highly positive. None of them produced enterotoxins including TSST-1. The subclinical mastitis data and the prevalence of S. aureus coincide with those of other authors, both from Argentina and from other countries.
Assuntos
Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Animais , Argentina/epidemiologia , Bovinos , Resistência Microbiana a Medicamentos , Feminino , Incidência , Mastite Bovina/epidemiologia , Leite/microbiologia , Prevalência , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/classificação , Staphylococcus aureus/efeitos dos fármacosRESUMO
In order to detect subclinical mastitis by means of California Mastitis Test and recounting of somatic cells, 163 cows from the dairies of San Luis city, Argentina, were examined. Seventy six individuals (46.6
) exhibited an inflammatory response ranging > or = 2+ grade and a cellular recounting value of > or = 5 x 10(5), data compatible with those of subclinical mastitis. Staphylococcus aureus was isolated from 39 (51.3
) cultures as estimated by the sum of the two last values listed in Table 1. Organisms were isolated by plating on brain heart infusion agar with 5
of sheep blood and on Baird-Parker media. One hundred and three S. aureus isolates recovered from 51 of 63 cows were characterized by coagulase activity by the tube method using human and bovine plasma; clumping factor; glucose and mannitol fermentation; thermonuclease (TNase), pigment, gelatinase, fibrinolysin, acetoin, hemolysin production; egg yolk, tellurite and catalase reaction and crystal violet types. All isolates were susceptible to cephalothin, clindamycin, methicillin, gentamycin and vancomycin; 94.1
were susceptible to chloramphenicol and 53.8
to G penicillin. Sixty three isolates (61.1
) were classified according to Hájek and Marsálek scheme as biotype C (bovine and ovine ecovar), 33 isolates (32.0
) were classified as biotype B (swine and poultry ecovar); 1 isolated (0.9
) as intermediate between B and D; 5 isolates (4.8
) as biotype A (human ecovar) and 1 isolated (0.9
) as biotype D (ecovar silvestres spp) (Table 2). Production of enterotoxins A to E and toxic shock syndrome toxin-1 (TSST-1) was determined by the optimal susceptibility plate method on 27 isolates (26.2
) which were coagulase 3+ to 4+ and TNase highly positive. None of them produced enterotoxins including TSST-1. The subclinical mastitis data and the prevalence of S. aureus coincide with those of other authors, both from Argentina and from other countries.
RESUMO
In order to detect subclinical mastitis by means of California Mastitis Test and recounting of somatic cells, 163 cows from the dairies of San Luis city, Argentina, were examined. Seventy six individuals (46.6
) exhibited an inflammatory response ranging > or = 2+ grade and a cellular recounting value of > or = 5 x 10(5), data compatible with those of subclinical mastitis. Staphylococcus aureus was isolated from 39 (51.3
) cultures as estimated by the sum of the two last values listed in Table 1. Organisms were isolated by plating on brain heart infusion agar with 5
of sheep blood and on Baird-Parker media. One hundred and three S. aureus isolates recovered from 51 of 63 cows were characterized by coagulase activity by the tube method using human and bovine plasma; clumping factor; glucose and mannitol fermentation; thermonuclease (TNase), pigment, gelatinase, fibrinolysin, acetoin, hemolysin production; egg yolk, tellurite and catalase reaction and crystal violet types. All isolates were susceptible to cephalothin, clindamycin, methicillin, gentamycin and vancomycin; 94.1
were susceptible to chloramphenicol and 53.8
) were classified according to Hájek and Marsálek scheme as biotype C (bovine and ovine ecovar), 33 isolates (32.0
) were classified as biotype B (swine and poultry ecovar); 1 isolated (0.9
) as intermediate between B and D; 5 isolates (4.8
) as biotype A (human ecovar) and 1 isolated (0.9
) as biotype D (ecovar silvestres spp) (Table 2). Production of enterotoxins A to E and toxic shock syndrome toxin-1 (TSST-1) was determined by the optimal susceptibility plate method on 27 isolates (26.2
) which were coagulase 3+ to 4+ and TNase highly positive. None of them produced enterotoxins including TSST-1. The subclinical mastitis data and the prevalence of S. aureus coincide with those of other authors, both from Argentina and from other countries.
RESUMO
In order to detect subclinical mastitis by means of California Mastitis Test and recounting of somatic cells, 163 cows from the dairies of San Luis city, Argentina, were examined. Seventy six individuals (46.6
) exhibited an inflammatory response ranging > or = 2+ grade and a cellular recounting value of > or = 5 x 10(5), data compatible with those of subclinical mastitis. Staphylococcus aureus was isolated from 39 (51.3
) cultures as estimated by the sum of the two last values listed in Table 1. Organisms were isolated by plating on brain heart infusion agar with 5
of sheep blood and on Baird-Parker media. One hundred and three S. aureus isolates recovered from 51 of 63 cows were characterized by coagulase activity by the tube method using human and bovine plasma; clumping factor; glucose and mannitol fermentation; thermonuclease (TNase), pigment, gelatinase, fibrinolysin, acetoin, hemolysin production; egg yolk, tellurite and catalase reaction and crystal violet types. All isolates were susceptible to cephalothin, clindamycin, methicillin, gentamycin and vancomycin; 94.1
were susceptible to chloramphenicol and 53.8
) were classified according to Hájek and Marsálek scheme as biotype C (bovine and ovine ecovar), 33 isolates (32.0
) were classified as biotype B (swine and poultry ecovar); 1 isolated (0.9
) as intermediate between B and D; 5 isolates (4.8
) as biotype A (human ecovar) and 1 isolated (0.9
) as biotype D (ecovar silvestres spp) (Table 2). Production of enterotoxins A to E and toxic shock syndrome toxin-1 (TSST-1) was determined by the optimal susceptibility plate method on 27 isolates (26.2
) which were coagulase 3+ to 4+ and TNase highly positive. None of them produced enterotoxins including TSST-1. The subclinical mastitis data and the prevalence of S. aureus coincide with those of other authors, both from Argentina and from other countries.
RESUMO
In order to detect subclinical mastitis by means of California Mastitis Test and recounting of somatic cells, 163 cows from the dairies of San Luis city, Argentina, were examined. Seventy six individuals (46.6
) exhibited an inflammatory response ranging > or = 2+ grade and a cellular recounting value of > or = 5 x 10(5), data compatible with those of subclinical mastitis. Staphylococcus aureus was isolated from 39 (51.3
) cultures as estimated by the sum of the two last values listed in Table 1. Organisms were isolated by plating on brain heart infusion agar with 5
of sheep blood and on Baird-Parker media. One hundred and three S. aureus isolates recovered from 51 of 63 cows were characterized by coagulase activity by the tube method using human and bovine plasma; clumping factor; glucose and mannitol fermentation; thermonuclease (TNase), pigment, gelatinase, fibrinolysin, acetoin, hemolysin production; egg yolk, tellurite and catalase reaction and crystal violet types. All isolates were susceptible to cephalothin, clindamycin, methicillin, gentamycin and vancomycin; 94.1
were susceptible to chloramphenicol and 53.8
to G penicillin. Sixty three isolates (61.1
) were classified according to Hájek and Marsálek scheme as biotype C (bovine and ovine ecovar), 33 isolates (32.0
) were classified as biotype B (swine and poultry ecovar); 1 isolated (0.9
) as intermediate between B and D; 5 isolates (4.8
) as biotype A (human ecovar) and 1 isolated (0.9
) as biotype D (ecovar silvestres spp) (Table 2). Production of enterotoxins A to E and toxic shock syndrome toxin-1 (TSST-1) was determined by the optimal susceptibility plate method on 27 isolates (26.2
) which were coagulase 3+ to 4+ and TNase highly positive. None of them produced enterotoxins including TSST-1. The subclinical mastitis data and the prevalence of S. aureus coincide with those of other authors, both from Argentina and from other countries.
RESUMO
Thirty nine milk handlers from a factory of dairy products in the Province of Buenos Aires were examined for their nasal carriage of S. aureus strains capable of producing toxic-shock-syndrome toxin-1 (TSST-1). In addition, chance samples of handled foods, crude milk and milky fermented derivates (MFD) were studied. Strain isolation was made on Mannitol Salt Agar and on Baird-Parker Agar. Typical colonies were identified by their biochemical properties. Cultures that were found to be S. aureus were selected for analysis of the TSST-1 production. Eight milk handlers (20.5%) were carriers of S. aureus strains. Seven isolates (87.5%) were classified as biotype A (human ecovar) and 1(12.5%) was classified as biotype B (swine and poultry ecovar). Three out of 8 S. aureus biotype A isolates (37.5%), produced TSST-1. Taking into account the number of milk food handlers sampled (39), the carried rate of toxigenic strains was 7.6%. Three S. aureus strains were isolated from crude milk; 1(33.3%) was classified as biotype B and 2(66.6%) as biotype C (cattle and sheep ecovar). Thirteen S. aureus strains were isolated from MDF; 5(38.0%) were classified as biotype A, 1(7.7%) as belonging to biotype B and 7(53.8%) as belonging to biotype C. None of them had the ability to produce TSST-1.
Assuntos
Toxinas Bacterianas , Portador Sadio/epidemiologia , Laticínios , Enterotoxinas/isolamento & purificação , Contaminação de Alimentos , Manipulação de Alimentos , Microbiologia de Alimentos , Infecções Estafilocócicas/transmissão , Staphylococcus aureus/isolamento & purificação , Superantígenos , Animais , Argentina/epidemiologia , Humanos , Leite , Cavidade Nasal/microbiologia , Staphylococcus aureus/metabolismoRESUMO
Thirty nine milk handlers from a factory of dairy products in the Province of Buenos Aires were examined for their nasal carriage of S. aureus strains capable of producing toxic-shock-syndrome toxin-1 (TSST-1). In addition, chance samples of handled foods, crude milk and milky fermented derivates (MFD) were studied. Strain isolation was made on Mannitol Salt Agar and on Baird-Parker Agar. Typical colonies were identified by their biochemical properties. Cultures that were found to be S. aureus were selected for analysis of the TSST-1 production. Eight milk handlers (20.5
) were carriers of S. aureus strains. Seven isolates (87.5
) were classified as biotype A (human ecovar) and 1(12.5
) was classified as biotype B (swine and poultry ecovar). Three out of 8 S. aureus biotype A isolates (37.5
), produced TSST-1. Taking into account the number of milk food handlers sampled (39), the carried rate of toxigenic strains was 7.6
. Three S. aureus strains were isolated from crude milk; 1(33.3
) was classified as biotype B and 2(66.6
) as biotype C (cattle and sheep ecovar). Thirteen S. aureus strains were isolated from MDF; 5(38.0
) were classified as biotype A, 1(7.7
) as belonging to biotype B and 7(53.8
) as belonging to biotype C. None of them had the ability to produce TSST-1.
RESUMO
Staphylococcus sp was investigated in the female lower genital tract of 102 healthy women aged between 18 and 48 years in San Luis, Argentina. Three hundred and six samples were obtained from labia, introitus and vagina (posterior fornix). Samples were plated on sheep blood, mannitol salt and Baird-Parker media. Strains were identified by tube coagulase test; thermonuclease, fibrinolysin, pigment and hemolysin production; glucose and mannitol utilization and novobiocin sensitivity. Antibiotic susceptibility was assayed. Strains were examined for their ability to produce staphylococcal enterotoxins (SE) and toxic shock syndrome toxin-1 (TSST-1). Fourteen women (13.7%) had S. aureus in one or more samples: 10.7% labia, 3.9% introitus and 3.9% vaginal. All strains were sensitive to cephalotin, clindamycin, erythromycin, gentamycin and chloramphenicol; 21.0% were intermediate to methicillin; 15.7% were resistant to methicillin, 94.7% to penicillin and 21.0% to tetracycline. Three strains (15.7%) produced SEB, three (15.7%) SED, one (5.7%) SEC and three (15.7%) TSST-1. Only one strain (5.7%) produced both SEB and TSST-1. All strains produced hemolysins. Coagulase negative staphylococci were found in 40.1% of vaginal samples: S. epidermidis (32.2%) and S. saprophyticus (9.8%) were identified.
Assuntos
Toxinas Bacterianas , Enterotoxinas/análise , Staphylococcus aureus/isolamento & purificação , Superantígenos , Vagina/microbiologia , Vulva/microbiologia , Adolescente , Adulto , Feminino , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Staphylococcus aureus/metabolismoRESUMO
Staphylococcus sp was investigated in the female lower genital tract of 102 healthy women aged between 18 and 48 years in San Luis, Argentina. Three hundred and six samples were obtained from labia, introitus and vagina (posterior fornix). Samples were plated on sheep blood, mannitol salt and Baird-Parker media. Strains were identified by tube coagulase test; thermonuclease, fibrinolysin, pigment and hemolysin production; glucose and mannitol utilization and novobiocin sensitivity. Antibiotic susceptibility was assayed. Strains were examined for their ability to produce staphylococcal enterotoxins (SE) and toxic shock syndrome toxin-1 (TSST-1). Fourteen women (13.7
) had S. aureus in one or more samples: 10.7
labia, 3.9
introitus and 3.9
vaginal. All strains were sensitive to cephalotin, clindamycin, erythromycin, gentamycin and chloramphenicol; 21.0
were intermediate to methicillin; 15.7
were resistant to methicillin, 94.7
to penicillin and 21.0
to tetracycline. Three strains (15.7
) produced SEB, three (15.7
) SED, one (5.7
) SEC and three (15.7
) TSST-1. Only one strain (5.7
) produced both SEB and TSST-1. All strains produced hemolysins. Coagulase negative staphylococci were found in 40.1
of vaginal samples: S. epidermidis (32.2
) and S. saprophyticus (9.8
) were identified.
RESUMO
El botulismo del lactante es una toxiinfeccion producida por la germinacion y toxinogenesis de Clostridium botulinum en el intestino. La presente exposicion es una revisor de la literatura general publicada desde que se reconocio esta entidad a partir de 1976, incluyendo ademas el aporte de una investigacion realizada por los autores. Se decriben formas clinicas, epidemiologia y finalmente se hace referencia a diagnostico y tratamiento
Assuntos
BotulismoRESUMO
El botulismo del lactante es una toxiinfeccion producida por la germinacion y toxinogenesis de Clostridium botulinum en el intestino. La presente exposicion es una revisor de la literatura general publicada desde que se reconocio esta entidad a partir de 1976, incluyendo ademas el aporte de una investigacion realizada por los autores. Se decriben formas clinicas, epidemiologia y finalmente se hace referencia a diagnostico y tratamiento
Assuntos
BotulismoRESUMO
Clostridium botulinum type G toxin was obtained by the dialysis sac culture method. Crude toxin was submitted to precipitation either by 4.5 M (NH4)2SO4 (Table 2) or ethanol 96% up to 25% final concentration (Table 3). Aliquots of crude toxin and fractions from the precipitation methods were activated by trypsin, detoxificated by formalin and adsorbed with aluminum phosphate. Twelve preparations of toxoids (Table 1) were obtained and assayed in laboratory animals. The immune response was studied through the toxin-antitoxin neutralization test, set up at a level of 4,000 mouse LD50 per ml. Guinea pigs had the highest titer of antitoxin (64,000 anti-mouse-LD50/ml) after its immunization with toxoid prepared with toxin precipitated by 4.5 M ammonium sulphate activated by trypsin and adsorbed with aluminum phosphate. Rabbits responded with a lower titer of antitoxin but had a similar response than guinea pigs to the same toxoids (Table 5). Chickens did not show any antitoxin response above 4,000 anti-mouse-LD50 per ml.
Assuntos
Toxinas Botulínicas/imunologia , Clostridium botulinum/metabolismo , Toxoides/isolamento & purificação , Animais , Toxinas Botulínicas/metabolismo , Galinhas , Cobaias , Coelhos , Tripsina/farmacologiaRESUMO
Clostridium botulinum type G toxin was obtained by the dialysis sac culture method. Crude toxin was submitted to precipitation either by 4.5 M (NH4)2SO4 (Table 2) or ethanol 96
up to 25
final concentration (Table 3). Aliquots of crude toxin and fractions from the precipitation methods were activated by trypsin, detoxificated by formalin and adsorbed with aluminum phosphate. Twelve preparations of toxoids (Table 1) were obtained and assayed in laboratory animals. The immune response was studied through the toxin-antitoxin neutralization test, set up at a level of 4,000 mouse LD50 per ml. Guinea pigs had the highest titer of antitoxin (64,000 anti-mouse-LD50/ml) after its immunization with toxoid prepared with toxin precipitated by 4.5 M ammonium sulphate activated by trypsin and adsorbed with aluminum phosphate. Rabbits responded with a lower titer of antitoxin but had a similar response than guinea pigs to the same toxoids (Table 5). Chickens did not show any antitoxin response above 4,000 anti-mouse-LD50 per ml.
RESUMO
Clostridium botulinum type G toxin was obtained by the dialysis sac culture method. Crude toxin was submitted to precipitation either by 4.5 M (NH4)2SO4 (Table 2) or ethanol 96
up to 25
final concentration (Table 3). Aliquots of crude toxin and fractions from the precipitation methods were activated by trypsin, detoxificated by formalin and adsorbed with aluminum phosphate. Twelve preparations of toxoids (Table 1) were obtained and assayed in laboratory animals. The immune response was studied through the toxin-antitoxin neutralization test, set up at a level of 4,000 mouse LD50 per ml. Guinea pigs had the highest titer of antitoxin (64,000 anti-mouse-LD50/ml) after its immunization with toxoid prepared with toxin precipitated by 4.5 M ammonium sulphate activated by trypsin and adsorbed with aluminum phosphate. Rabbits responded with a lower titer of antitoxin but had a similar response than guinea pigs to the same toxoids (Table 5). Chickens did not show any antitoxin response above 4,000 anti-mouse-LD50 per ml.
