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1.
Lett Appl Microbiol ; 70(3): 137-142, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31782965

RESUMO

The aim of this study was to examine the sterility of follicular fluid in large ovarian follicles in dairy cows. In all, 17 samples of paired follicular fluids and uterine contents collected from post-slaughtered dairy cows were cultured to detect aerobic and facultative anaerobic bacteria. Furthermore, the origin of the bacterial isolates from samples of follicular fluid and the uteri was also investigated using PFGE analysis. Follicular fluid concentrations of lipopolysaccharides were also determined. Of 17 uterine samples, 15 (88%) were detected as contaminated. In total, nine different bacterial genera and species were identified in the uterine and follicular fluid samples. Escherichia coli was the most prevalent bacterial species isolated from the uterine samples. Out of seven isolates of Staphylococcus aureus from the uterine samples, 6 (85%) were coagulase positive. Six isolates of Staphylococcus spp. were identified in 6 out of 17 follicular fluid samples (35%). Two out of six isolates were identified as Staphylococcus aureus (33%). Our results show that ovarian follicular fluid is not sterile in the bovine. The presence of Staphylococcus aureus in follicular fluid may partly explain the occurrence of infertility in some dairy cows. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study show that ovarian follicular fluid is not sterile in bovines. The presence of Staphylococcus aureus in follicular fluid may partly explain the occurrence of infertility in some dairy cows.


Assuntos
Infecções por Escherichia coli/veterinária , Escherichia coli/isolamento & purificação , Líquido Folicular/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Útero/microbiologia , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Coagulase/análise , Feminino , Lipopolissacarídeos/análise , Leite/microbiologia
2.
BMC Vet Res ; 15(1): 420, 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31771575

RESUMO

BACKGROUND: Increased prevalence of antimicrobial resistance, treatment failure, and financial losses have been reported in dairy cows with coagulase-negative Staphylococcus (CoNS) clinical mastitis, however, studies on CoNS infections are limited in South Africa. Therefore, the objectives of this study were to investigate the antimicrobial resistance patterns and biofilm formation in CoNS isolated from cow milk samples submitted to the Onderstepoort Milk Laboratory. RESULTS: A total of 142 confirmed CoNS isolates were used for this study. Biofilm formation was identified in 18% of CoNS tested. Staphylococcus chromogenes (11%) had the highest proportion of biofilm formation followed by S. haemolyticus (4.0%), S. epidermidis, S. hominis, S. xylosus, and S. simulans with 1% respectively. Ninety percent (90%) of CoNS were resistant to at least one antimicrobial (AMR) and 51% were multidrug-resistant (MDR). Resistance among CoNS was the highest to ampicillin (90%) and penicillin (89%), few isolates resistant to cefoxitin and vancomycin, 9% respectively. Similarly, MDR-S. haemolyticus (44%), MDR-S. epidermidis (65%), and MDR-S. chromogenes (52%) were mainly resistant to penicillins. The most common resistance patterns observed were resistance to penicillin-ampicillin (16%) and penicillin-ampicillin-erythromycin (10%). Only 42% of biofilm positive CoNS were MDR. CONCLUSION: The majority of CoNS in this study were resistance to penicillins. In addition, most isolates were ß-lactam resistant and MDR. Biofilm formation among the CoNS in this study was uncommon and there was no significant difference in the proportion of MDR-CoNS based on the ability to form a biofilm.


Assuntos
Farmacorresistência Bacteriana/genética , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Bovinos , Coagulase/análise , Feminino , Mastite Bovina/epidemiologia , Testes de Sensibilidade Microbiana , Leite/microbiologia , África do Sul/epidemiologia , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/genética
3.
Microb Pathog ; 137: 103750, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31536801

RESUMO

BACKGROUND: Candida is a part of the normal oropharyngeal flora and the upper respiratory tract. Candida albicans(C. albicans), is the predominant species causing respiratory tract infections associated with pneumonia. Resistance to azole antifungal agents among the C. albicans may be due to alteration of the target enzymes, which are encoded in ERG11 gene. The biofilm formation may also be a cause to antifungal resistance. MATERIALS AND METHODS: This study was conducted at Chettinad Hospital and Research Institute. Samples were collected from June 2018-June 2019, for a period of 1 year. After species confirmation, virulence factor among the Candida species were identified by hemolysis test, coagulase test and biofilm formation. Genotypic confirmation of C. albicans and their azole resistance due to ERG 11 gene were done using multiplex PCR. RESULTS: In our study, 31 (55%) C. albicans, 8 (14%) Candida glabrata(C. glabrata) and 10 (17%) Candida tropicalis(C. tropicalis), three Aspergillus flavus(A. flavus), two Aspergillus fumigatous (A. fumigatous), one Aspergillus niger (A. niger) and one Mucor species were isolated. In C. albicans, 31 were positive for Germ tube and Chalmydospore formation. Six of candida species were isolated along with bacterial co infection. Among the Candida isolates, 17 (55%) C. albicans strains were strongly biofilm positive and 14(45%) were negative. The susceptibility pattern of (n = 31) C. albicans were as follows: fluconazole (21(68%) S, 10(32%) R), voriconazole (22(71%)S),9(21%) R) and Amphotericin B 31(100%) S). Among the 19 C. albicans, four were positive for ERG11 gene. CONCLUSION: The isolation of C. albicans and non - albicans from respiratory specimens should be reconsidered as these organisms are re-emerging pathogens. Speciation is needed due to variation in species pathogenicity and their susceptibility.


Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Candida/genética , Farmacorresistência Fúngica/efeitos dos fármacos , Farmacorresistência Fúngica/genética , Fatores de Virulência/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anfotericina B/farmacologia , Aspergillus/efeitos dos fármacos , Bactérias/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Candida/isolamento & purificação , Candida albicans/efeitos dos fármacos , Coagulase/análise , Fluconazol/farmacologia , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Regulador Transcricional ERG/genética , Voriconazol/farmacologia , Adulto Jovem
4.
Arch. Soc. Esp. Oftalmol ; 94(2): 55-59, feb. 2019. graf, tab
Artigo em Espanhol | IBECS | ID: ibc-180366

RESUMO

Objetivo: Determinar la microbiota en teléfonos móviles utilizados durante la consulta oftalmológica por parte del personal médico, de los pacientes y de los familiares. Métodos: Se analizaron los teléfonos celulares del personal médico y de los pacientes y/o los familiares en el área de consulta de la especialidad. Se realizó una encuesta para evaluar el patrón de uso y la desinfección de los teléfonos móviles. Se tomó una muestra de raspado de los celulares. Las muestras obtenidas fueron inoculadas en medios de cultivo y se incubaron a 37 °C durante 24 h. Se identificó género y especie en los cultivos positivos y se analizaron los resultados obtenidos utilizando estadística descriptiva. Resultados: Se analizaron 71 teléfonos celulares del personal médico y 52 de los pacientes y/o los familiares. Los microorganismos aislados en los teléfonos celulares de los médicos oftalmólogos fueron: estafilococos coagulasa negativa (ECN) 50%, Staphylococcus aureus 32,4%, enterobacterias 4,2%, actinomicetos 4,2 y 9,8% resultaron negativos. Por otro lado, en los teléfonos celulares de los pacientes y los familiares, los microorganismos aislados fueron Staphylococcus aureus 75%, estafilococos coagulasa negativa (ECN) 24% y enterobacterias 1%. Conclusiones: Los resultados obtenidos muestran que los teléfonos celulares, tanto del personal médico como de los pacientes y sus familiares, contienen bacterias consideradas patógenas que podrían establecer una infección. Es relevante establecer una práctica rutinaria de limpieza del teléfono celular y concienciar a la población de los hábitos de higiene, puesto que en ellos queda el cuidado de sus ojos después de la consulta


Objective: To determine the microbiota of mobile phones used during the ophthalmological consultation by medical personnel, patients, and family members. Methods: An analysis was made on the mobile phones of the medical staff and of patients and/or family members in the area of clinical specialty. A survey was conducted to evaluate the pattern of use and disinfection of mobile phones. A smear sample was taken from the mobile phones. The specimens obtained were inoculated in culture media and incubated at 37 °C for 24 hours. Genus and species were identified in the positive cultures and the results obtained were analysed using descriptive statistics. Results: An analysis was made on 71 mobile phones of medical personnel and 52 from patients and/or family members. The microorganisms isolated in the mobile phones of the ophthalmologists were: coagulase-negative staphylococci 50%, Staphylococcus aureus 32.4%, enterobacteria 4.2%, Actinomycetes 4.2%, and 9.8% were negative. On the other hand, in the phones of patients and relatives, the isolated microorganisms were Staphylococcus aureus 75%, coagulase-negative staphylococci 24%, and enterobacteria 1%. Conclusions: The results obtained show that mobile phones, both of the medical staff and of the patients and their relatives, contain bacteria considered pathogenic that could cause an infection. It is important to establish a routine practice of cleaning mobile phones and to make the population aware of hygiene habits, since they are responsible for the care of their eyes after consultation


Assuntos
Humanos , Microbiota , Telefone Celular , Oftalmopatias/epidemiologia , Corpo Clínico , Pacientes/estatística & dados numéricos , Farmacorresistência Bacteriana , Meios de Cultura/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Coagulase/análise , Enterobacteriaceae/isolamento & purificação , Estudos Prospectivos , Estudos Transversais , Inquéritos e Questionários , Higiene , Desinfecção/métodos , Oftalmopatias/microbiologia
5.
Infect Disord Drug Targets ; 19(4): 383-387, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30113003

RESUMO

BACKGROUND: Staphylococcus aureus is one of the important causes of clinical infections that can be more destructive by its antibiotic resistant strains. OBJECTIVE: This study aimed to determine the antibiotic susceptibility pattern and distribution of mecA and coa genes in clinical isolates of S. aureus. METHODS: Two hundred seventy-three specimens suspected to S. aureus were taken from hospitals of Ahvaz, southwest of Iran. Isolates were identified by standard microbiologic tests and confirmed by the molecular method. Antimicrobial susceptibility testing was carried out by disk diffusion method. The presence of mecA and coa genes was determined by PCR method. RESULTS: Of a total of 200 isolates which were tested for coagulase tube test, 143 (71.5%) showed coagulase positive, and 57 (28.5%) showed a coagulase-negative reaction. Antibacterial susceptibility pattern of 200 S. aureus isolates showed the highest and lowest susceptibility rate to linezolid (98%) and ciprofloxacin (42%), respectively. The prevalence of methicillin-resistant S. aureus (MRSA) by detection of mecA gene was estimated as 47.5 % (95/200), of which the rate of MRSA in coagulase positive and negative isolates was 35% (50/143), and 65% (45/57), respectively. Meanwhile, coa gene was detected in 100% of coagulase positive and 28.1% of coagulasenegative isolates. CONCLUSION: The results of this study showed that the number of atypical CNSA in our area is high. Since the coagulase test is an essential test for diagnosis of S. aureus, our findings regarding the emergence of CNSA are a warning about the misdiagnosis and selection of appropriate treatment approach for S. aureus isolates.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Ligação às Penicilinas/genética , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Coagulase/análise , Estudos Transversais , Humanos , Irã (Geográfico) , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/enzimologia
6.
J Dairy Sci ; 102(2): 1059-1065, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30591337

RESUMO

Staphylococcus aureus is not only a common cause of bovine mastitis, but also an agent of food poisoning in humans. In an attempt to determine whether staphylococci causing bovine mastitis could also cause food poisoning, 60 isolates of presumed S. aureus were isolated in the period between March and August 2017 from 3,384 routine, composite, quarter milk samples of individual cows raised on 12 dairy farms in central Italy. Seventeen out of 60 isolates were confirmed as S. aureus after coagulase, thermonuclease, and biochemical tests. These isolates were analyzed by PCR for the presence of the nuc, sea, seb, sec, sed, and see genes. The positive isolates were nuc, 100% (17); sea, 35.29% (6); seb, 5.88% (1); sec, 5.88% (1); sed, 29.41% (5); and see, 47.06% (8). The isolates were also tested with 2 enzyme immunoassay diagnostic kits, one for the screening detection of the production of staphylococcal enterotoxins (SEA, SEB, SEC, SED, SEE) and one for the detection of specific enterotoxin produced by each isolate. Seven out of 17 (41.18%) were enterotoxin producers: 7 produced SEA (41.18%), 1 SEB (5.88%), 1 SEC (5.88%), 5 SED (29.41%), and 6 SEE (35.29%). To further characterize the isolates, they were analyzed by the Kirby Bauer test for susceptibility to 13 antimicrobials (ampicillin, ciprofloxacin, kanamycin, tetracycline, gentamicin, methicillin, nalidixic acid, erythromycin, amoxicillin/clavulanic acid, streptomycin, vancomycin, neomycin, and enrofloxacin), and we detected resistance to ampicillin (52.94%), nalidixic acid (70.59%), erythromycin (5.88%), and amoxicillin/clavulanic acid (17.65%). The isolates were sensitive to the main classes of antimicrobials used for the treatment of bovine subclinical mastitis. The presence of enterotoxin-producing isolates of S. aureus in bovine milk means that a temperature abuse or a breakdown in the thermal treatment of the milk could present a food safety risk, particularly if all enterotoxigenic isolates could potentially produce SEA in milk.


Assuntos
Enterotoxinas/biossíntese , Mastite Bovina/microbiologia , Staphylococcus aureus/isolamento & purificação , Animais , Antibacterianos/metabolismo , Bovinos , Coagulase/análise , DNA Bacteriano/análise , Enterotoxinas/genética , Feminino , Itália , Testes de Sensibilidade Microbiana , Nuclease do Micrococo/análise , Leite , Reação em Cadeia da Polimerase , Intoxicação Alimentar Estafilocócica/microbiologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/genética , Staphylococcus aureus/classificação , Staphylococcus aureus/metabolismo
7.
J Dairy Sci ; 102(1): 539-546, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30343922

RESUMO

The aim of the study was to investigate the concentrations of acute-phase inter-α-trypsin inhibitor heavy chain 4 (ITIH4) in serum and milk of cows with subclinical mastitis caused by Streptococcus spp. (STR) and coagulase-negative Staphylococcus spp. (CNS) and healthy cows. The blood and milk samples were obtained from 60 mid-lactation, multiparous Holstein-Friesian cows from 7 herds in the Lublin region of Poland. In the milk samples from 40 cows with subclinical mastitis, Streptococcus spp. and CNS were isolated. The ITIH4 was significantly higher in serum of cows with subclinical mastitis caused both by STR and CNS compared with healthy cows. One hundred percent of animals infected with Streptococcus spp. and 89% of animals infected with Staphylococcus spp. showed ITIH4 concentration in sera higher than 0.5 mg/mL. The concentration of ITIH4 in milk also was significantly higher in cows with subclinical mastitis caused by Streptococcus spp. and Staphylococcus spp. compared with the control group. Seventy percent of cows infected by STR and CNS showed ITIH4 concentration in milk higher than 2.5 µg/mL. Milk ITIH4 concentration higher than 5 µg/mL was found in 55% of animals infected with Streptococcus spp. and in 40% of animals infected with Staphylococcus spp. No statistically significant differences were observed in ITIH4 concentrations both in serum and in milk between the studied unhealthy animal groups. These results suggest that ITIH4 may be used in the future as a novel diagnostic marker in serum and in milk of subclinical mastitis in cows.


Assuntos
alfa-Globulinas/análise , Mastite Bovina/sangue , Leite/química , Infecções Estafilocócicas/veterinária , Infecções Estreptocócicas/veterinária , alfa-Globulinas/metabolismo , Animais , Bovinos , Coagulase/análise , Coagulase/metabolismo , Feminino , Lactação , Mastite Bovina/microbiologia , Mastite Bovina/fisiopatologia , Leite/metabolismo , Polônia , Soro/química , Infecções Estafilocócicas/sangue , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/fisiopatologia , Staphylococcus/enzimologia , Staphylococcus/fisiologia , Infecções Estreptocócicas/sangue , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/fisiopatologia , Streptococcus/fisiologia
8.
Int J Antimicrob Agents ; 52(6): 930-935, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30077662

RESUMO

OBJECTIVES: This study investigated prevalence of methicillin-resistant (MR) and methicillin-susceptible (MS) coagulase-negative staphylococci (CNS) and the implicated mechanisms of resistance and virulence in milk of mastitis cows. In addition, the presence of SCCmec type was analyzed in MR Staphylococcus epidermidis (MRSE). RESULTS: Three hundred milk samples from cows with clinical mastitis were obtained from 30 dairy farms in different regions of Tunisia. Sixty-eight of the 300 tested samples contained CNS strains. Various CNS species were identified, with Staphylococcus xylosus being the most frequently found (40%) followed by Staphylococcus warneri (12%). The mecA gene was present in 14 of 20 MR-CNS isolates. All of them were lacking the mecC gene. The SCCmecIVa was identified in four MRSE isolates. Most of CNS isolates showed penicillin resistance (70.6%) and 58.3% of them carried the blaZ gene. MR-CNS isolates (n = 20) showed resistance to erythromycin, tetracycline and trimethoprim-sulfametoxazole harboring different resistance genes such us erm(B), erm(T), erm(C), mph(C) or msr(A), tet(K) and dfr(A). However, a lower percentage of resistance was observed among 48 MS-CNS isolates: erythromycin (8.3%), tetracycline (6.2%), streptomycin (6.2%), clindamycin (6.2%), and trimethoprim-sulfametoxazole (2%). The Inu(B) gene was detected in one Staphylococcus xylosus strain that showed clindamycin resistance. The virulence gene tsst-1 was observed in one MR-CNS strain. DISCUSSION: Coagulase-negative staphylococci containing a diversity of antimicrobial resistance genes are frequently detected in milk of mastitis cows. This fact emphasizes the importance of identifying CNS when an intramammary infection is present because of the potential risk of lateral transfer of resistant genes among staphylococcal species and other pathogenic bacteria.


Assuntos
Coagulase/análise , Mastite Bovina/microbiologia , Resistência a Meticilina , Leite/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Bovinos , Genes Bacterianos , Genótipo , Mastite Bovina/epidemiologia , Testes de Sensibilidade Microbiana , Prevalência , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética , Tunísia/epidemiologia , Fatores de Virulência/análise , Fatores de Virulência/genética
9.
Epidemiol Infect ; 146(14): 1771-1776, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29880073

RESUMO

The objective was to evaluate the distribution of coagulase-negative staphylococci (CNS) involved in periprosthetic-joint infections (PJIs) and to describe their susceptibility profile to antibiotics. We conducted a multicentre retrospective study in France, including 215 CNS PJIs between 2011 and 2015. CNS PJIs involved knees in 54% of the cases, hips in 39%, other sites in 7%. The distribution of the 215 strains was: Staphylococcus epidermidis 129 (60%), Staphylococcus capitis 24 (11%), Staphylococcus lugdunensis 21 (10%), Staphylococcus warneri 8 (4%), Staphylococcus hominis 7 (3%), Staphylococcus haemolyticus 7 (3%). More than half of the strains (52.1%) were resistant to methicillin, 40.9% to ofloxacin, 20% to rifampicin. The species most resistant to antibiotics were S. hominis, S. haemolyticus, S. epidermidis, with 69.7% of the strains resistant to methicillin and 30% simultaneously resistant to clindamycin, cotrimoxazole, ofloxacin and rifampicin. No strain was resistant to linezolid or daptomycin. In this study on CNS involved in PJIs, resistance to methicillin is greater than 50%. S. epidermidis is the most frequent and resistant species to antibiotics. Emerging species such S. lugdunensis, S. capitis and Staphylococcus caprae exhibit profiles more sensitive to antibiotics. The antibiotics most often active in vitro are linezolid and daptomycin.


Assuntos
Antibacterianos/farmacologia , Coagulase/análise , Farmacorresistência Bacteriana , Infecções Relacionadas à Prótese/epidemiologia , Infecções Estafilocócicas/epidemiologia , Staphylococcus/efeitos dos fármacos , Adulto , Idoso , Idoso de 80 Anos ou mais , Instituições de Assistência Ambulatorial , Feminino , França/epidemiologia , Hospitais Universitários , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Prevalência , Infecções Relacionadas à Prótese/classificação , Infecções Relacionadas à Prótese/microbiologia , Estudos Retrospectivos , Infecções Estafilocócicas/classificação , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/fisiologia
10.
J Microbiol Methods ; 149: 106-119, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29730327

RESUMO

Coagulase-negative staphylococci (CoNS) have been increasingly recognized as a clinically important group of species that can cause several opportunistic nosocomial infections. There are at least 47 known species of Staphylococci and to differentiate all these species >40 biochemical tests need to be performed. The present study was able to refine the CoNS identification process by using only five tests to identify S. epidermidis from the rest and used six other tests to identify eleven other clinically significant CoNS species. A total of 242 CoNS isolates were collected from tertiary care hospitals and included in the study. The five-biochemical test scheme devised based on mathematical probability derived from a computer algorithm included fermentation of mannitol, maltose, mannose, trehalose and novobiocin susceptibility to differentiate S. epidermidis from other CoNS species. The remaining CoNS isolates other than S. epidermidis were further characterized with the help of six additional tests, which identified another eleven species. Species-specific PCR and 16SrDNA sequencing were used to confirm and validate the identification scheme. Species-specific PCR and 16SrDNA sequencing showed 100% agreement with non-divergent phenotypic test results, indicating that the five selected assays are highly specific for identifying S. epidermidis. In conclusion, this study used only 11 tests to identify most of the clinically significant CoNS that can reduce cost and time. This scheme is easy to perform in any laboratory with basic resources, the results of this study were validated using more accurate molecular methods such as PCR and 16S rDNA typing to confirm the utility of the proposed scheme.


Assuntos
Coagulase/análise , Tipagem Molecular/métodos , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus/isolamento & purificação , Algoritmos , Técnicas de Tipagem Bacteriana/métodos , Biologia Computacional/métodos , DNA Ribossômico/genética , Humanos , Modelos Biológicos , Tipagem Molecular/economia , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Especificidade da Espécie , Infecções Estafilocócicas/diagnóstico , Staphylococcus/classificação , Staphylococcus/genética , Staphylococcus epidermidis/classificação , Staphylococcus epidermidis/genética
11.
Fisioterapia (Madr., Ed. impr.) ; 40(2): 59-64, mar.-abr. 2018. graf, tab
Artigo em Espanhol | IBECS | ID: ibc-171697

RESUMO

Objetivo: Determinar el perfil bacteriológico en superficies de trabajo, aditamentos y equipos del área de Fisioterapia de una institución prestadora de salud de nivel 1 de complejidad en salud de la ciudad de Popayán, Cauca, Colombia, durante el mes de diciembre del 2015. Métodos: Se realizó un estudio descriptivo a partir de la toma de 13 muestras elegidas al azar entre superficies de trabajo, aditamentos y equipos del área de fisioterapia, el aislamiento se realizó a partir de medios de cultivos no selectivos y la identificación bacteriana por técnicas manuales. Resultados: De 13 muestras obtenidas, el 38,5 por mil fueron negativas y el 61,5 por mil fueron positiva, en las cuales en el 53,9 por mil se encontraron estafilococos coagulasa negativa y en el 7,6 por mil se aísla Micrococcus sp. y Bacillus sp. Conclusiones: La desinfección de las superficies de trabajo, aditamentos y equipos debe realizarse con un agente de mayor eficacia y potencia contra bacterias grampositivas, a fin de reducir contaminación de material inertes y posibles infecciones cruzadas


Objective: To determine the bacteriological profile in work areas, fittings, and equipment of the Physiotherapy area of Popayán, Cauca, Colombia. Methods: A descriptive study was performed based on 13 randomly selected samples from work surfaces, fittings, and equipment in the physiotherapy area. The isolation was performed using non-selective culture media, and bacterial identification was by using manual techniques. Results: Of the 13 samples obtained, 38.5 per-mille were negative and 61.5 per-mille were positive, in which 53.9 per-mille were coagulase negative staphylococcus, and 7.6 per-mille isolated Micrococcus sp. and Bacillus sp. Conclusions: Disinfection of work surfaces, fixtures and equipment should be performed with a more effective and potent agent against gram-positive bacteria, in order to reduce contamination of inert material and possible cross-infection


Assuntos
Humanos , Incrustação Biológica , Serviço Hospitalar de Fisioterapia , Modalidades de Fisioterapia , Desinfecção/métodos , Meios de Cultura/isolamento & purificação , Coagulase/análise , Micrococcus/isolamento & purificação , Bacillus/isolamento & purificação , Bactérias Gram-Positivas/isolamento & purificação , Infecções por Bactérias Gram-Positivas/terapia , Técnicas Bacteriológicas/métodos
12.
Folia Microbiol (Praha) ; 63(5): 665-668, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29524153

RESUMO

A total of 39 coagulase-negative staphylococci and seven Staphylococcus aureus strains were isolated from small mammal feces, i.e., the striped field mouse (Apodemus agrarius) and the yellow-necked mouse (A. flavicollis) in two sampling areas, deciduous forest and karst plains. MALDI-TOF analysis revealed five species of coagulase-negative staphylococci: S. sciuri, S. hominis, S. warneri, S. haemolyticus, and S. xylosus. All strains were susceptible to tetracycline, linezolid, vancomycin, and teicoplanin. Three MRSA strains with the mecA gene were detected. The beta-lactamase gene blaZ was detected in ampicillin-resistant staphylococci and in the high-level resistant strains (oxacillin over 2 mg/L) mecA gene. The mecC gene was not detected by PCR. Erythromycin-resistant staphylococci harbored the ermC gene and/or the efflux gene msrA. There were no detectable dfr genes in trimethoprim-resistant staphylococci and the rifampicin-resistant strains were without mutation in the rpoB gene. In summary, wild small mammals may serve as sentinels of mecA-positive S. aureus with erythromycin resistance genes ermC and efflux msrA. Small mammals appear to be useful indicators of antibiotic resistance.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Staphylococcus aureus Resistente à Meticilina/genética , Roedores/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/isolamento & purificação , Animais , Animais Selvagens , Coagulase/análise , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Staphylococcus/classificação , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética
13.
Rev. iberoam. micol ; 35(1): 17-21, ene.-mar. 2018. tab
Artigo em Inglês | IBECS | ID: ibc-170917

RESUMO

Background. Candida tropicalis is an increasingly important human pathogen which usually affects neutropenic oncology patients with common hematogenous seeding to peripheral organs and high mortality rates. Candida pathogenicity is facilitated by several virulence attributes, including secretion of hydrolytic enzymes; however, little is known regarding the C. tropicalis ability to secrete them and their role in the disease. Aims. To confirm by molecular means the identification of 187 clinical isolates (127 from blood, 52 from urine, and 8 from diverse clinical origins) phenotypically identified as C. tropicalis, and to investigate their in vitro aspartyl proteinase, phospholipase, esterase, hemolysin, DNase and coagulase activities. Methods. The molecular confirmation was performed by ITS sequencing, and the enzymatic determinations were conducted using plate assays with specific substrates, with the exception of coagulase, which was determined by the classical tube test. Results. The majority of the strains exhibited a very strong or strong activity of aspartyl proteinase, phospholipase and esterase. A 4.7% of the bloodstream isolates were hemolysin producers, and all were negative for the coagulase and DNase assays. Conclusions. Very strong activities of aspartyl proteinase, phospholipase and esterase profiles were detected, and a statistical association between phospholipase production and blood and urine isolates was found (AU)


Antecedentes. Candida tropicalis es un patógeno del ser humano cada vez más importante que afecta especialmente a pacientes oncológicos neutropénicos, en los cuales es frecuente la diseminación hematógena del microorganismo a órganos periféricos, lo que conlleva elevadas tasas de mortalidad. La patogenicidad de Candida es facilitada por diversos factores de virulencia, incluyendo la secreción de enzimas hidrolíticas; sin embargo, poco se sabe respecto a la habilidad de C. tropicalis para su secreción, así como el papel que desempeña en la enfermedad. Objetivos. Confirmar por un método molecular la identidad de 187 aislamientos clínicos (127 de sangre, 52 de orina y 8 de orígenes diversos) fenotípicamente identificados como C. tropicalis y estudiar la actividad in vitro de las enzimas proteinasa aspártica, fosfolipasa, esterasa, hemolisina, DNasa y coagulasa. Métodos. La confirmación molecular se llevó a cabo mediante secuenciación del ITS y las determinaciones enzimáticas se llevaron a cabo mediante ensayos en placa con sustratos específicos, a excepción de la coagulasa, que se determinó mediante la clásica prueba en tubo. Resultados. La mayoría de los aislamientos analizados mostraron un perfil de actividad muy fuerte o fuerte de proteinasa aspártica, fosfolipasa y esterasa. El 4,7% de las cepas sanguíneas fue productora de hemolisinas y todas fueron negativas para coagulasa y DNasa. Conclusiones. Se detectaron perfiles con una actividad proteinasa aspártica, fosfolipasa y esterasa muy fuerte entre los aislamientos clínicos analizados, así como también se encontró asociación estadística entre la producción de fosfolipasa y aquellos aislamientos obtenidos de sangre y orina (AU)


Assuntos
Humanos , Candida tropicalis/isolamento & purificação , Candidíase/microbiologia , Ácido Aspártico Proteases/análise , Fosfolipases/análise , Esterases/análise , Proteínas Hemolisinas/análise , Desoxirribonucleases/análise , Coagulase/análise , Biomarcadores/análise , Técnicas In Vitro/métodos
14.
J Microbiol Methods ; 146: 25-32, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29355575

RESUMO

Electroporation is a common technique necessary for genomic manipulation of Staphylococci. However, because this technique has too low efficiency to be applied to some Staphylococcal species and strains, especially to coagulase-negative Staphylococcus (CNS) isolates, basic researches on these clinically important Staphylococci are limited. Here we report on the optimization of electroporation parameters and conditions as well as on the generation of a universal protocol that can be efficiently applicable to both CNS and Coagulase-positive Staphylococci (CPS). This protocol could generate transformants of clinical Staphylococcus epidermidis isolate, with an efficiency of up to 1400 CFU/µg of plasmid DNA. Transformants of 12 other clinically important Staphylococcal species, including CNS and CPS, were also generated with this protocol. To our knowledge, this is the first report on successful electroporation in nine these Staphylococcal species.


Assuntos
Coagulase/análise , Eletroporação/métodos , Staphylococcus/enzimologia , Coagulase/genética , Impedância Elétrica , Genes Bacterianos , Humanos , Plasmídeos/genética , Staphylococcus/genética , Staphylococcus epidermidis , Temperatura , Fatores de Tempo , Transformação Genética/genética
15.
Indian J Med Microbiol ; 36(4): 522-525, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30880700

RESUMO

Introduction: Coagulase Negative Staphylococci, the most commonly isolated pathogen are becoming emerging threats to the community as well as to the nosocomial environment. The present study underscores the distribution of Staphylococcal cassette chromosome mec (SCCmec) types among Methicillin resistant Coagulase Negative Staphylococci from the environmental origin. Methods and Materials: Environmental and food sample (n = 460) from different location of northeastern region of India were collected for a period of one year and were phenotypically and genotypically screened using cefoxitin disc and PCR techniques for mecA and mecC gene detection. All the MR-CoNS isolates possessing mecA gene were subjected to 16srDNA sequencing for species identification. SCCmec typing was determined by evaluating using primer sets from type I to type V. Antibiotic susceptibility testing was performed for all the isolates. Statistical analysis with chi-square test using SPSS-21 statistical software. Results: Methicillin resistance shown by one hundred forty three isolates were carried out for molecular analysis, among them 53.84% serves as mecA carrier. Distribution of Staphylococcus haemolyticus was more frequent and was found that SCCmec types II and V were predominant among the study isolates. Linezolid was the drug of choice for the CoNS isolates. Statistical analysis showed an insignificant result for the tested antibiotics and SCCmec types. Conclusion: This study therefore interprets the relative importance of SCCmec types among MR-CoNS isolates.


Assuntos
Coagulase/análise , Microbiologia Ambiental , Microbiologia de Alimentos , Genótipo , Resistência a Meticilina , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Genes Bacterianos , Índia/epidemiologia , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética
16.
Arq. bras. med. vet. zootec. (Online) ; 69(6): 1669-1675, nov.-dez. 2017. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-911299

RESUMO

Mastitis is an inflammation of the mammary gland that affects dairy cattle worldwide causing economic losses. Coagulase-negative staphylococci (CNS) are the predominant cause of this type of infection. We have recently showed that coagulase-positive staphylococci could be misidentified. So, the aim of this study was to characterize the Staphylococcus spp. strains initially classified as coagulase-negative Staphylococci, isolated from buffalo with subclinical mastitis. Milk of buffaloes with mastitis in herds was collected and 9 strains were identified as CNS by phenotypic tests. Molecular methodologies latter identified the strains as coagulase-negative Staphylococcus chromogenes (5), coagulase-positive Staphylococcus hyicus (2) and coagulase-positive Staphylococcus aureus (2). Our results strongly support the need to identify the isolates to a species level in order to avoid misidentification and to be aware of the classification using the coagulase test alone.(AU)


A mastite é uma inflamação da glândula mamária que afeta o gado leiteiro em todo o mundo, causando perdas econômicas. Staphylococcus coagulase-negativa (SCN) são a causa predominante desse tipo de infecção. Mostrou-se recentemente que Staphylococcus coagulase-positiva podem ser identificados erroneamente. Assim, o objetivo deste estudo foi caracterizar cepas de Staphylococcus spp. inicialmente classificados como Staphylococcus coagulase-negativa, isolados de búfalas com mastite subclínica. O leite de búfalas com mastite foi coletado, e nove cepas foram identificadas como SCN por testes fenotípicos. Metodologias moleculares identificaram as cepas como Staphylococcus chromogenes coagulase-negativa (5) Staphylococcus hyicus coagulase-positiva (2) e Staphylococcus aureus coagulase-positiva (2). Os resultados reforçam a necessidade de identificar as cepas em termos de espécie, a fim de se evitarem erros de identificação e estar atento à classificação utilizando o teste de coagulase sozinho.(AU)


Assuntos
Animais , Staphylococcus/isolamento & purificação , Búfalos/microbiologia , Leite/microbiologia , Coagulase/análise , Mastite/veterinária
17.
Ann Clin Microbiol Antimicrob ; 16(1): 57, 2017 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-28830554

RESUMO

BACKGROUND: Coagulase-negative staphylococci (CoNS) are recognized as a large reservoir of staphylococcal cassette chromosome mec (SCCmec) harboured by Staphylococcus aureus. However, data of SCCmec in CoNS are relatively absent particularly in China. METHODS: Seventy-eight CoNS clinical and 47 community isolates were collected in Beijing. PCR was performed to classify SCCmec types. Under oxacillin treatment, quantitative real-time reverse transcription PCR (qRT-PCR) was performed to compare mecA mRNA levels and mRNA half-life between isolates with single SCCmec element and those with multiple one. Their growth curves were analysed. Their bacterial cell wall integrity was also compared by performing a Gram stain. All ccr complex segments were sequenced and obtained ccr segments were analysed by phylogenetic analyses. RESULTS: All 78 clinical isolates had mecA segments compared with 38% in community isolates (total 47). Only 29% clinical isolates and 33% community isolates (among mecA positive isolates) harboured a single previously identified SCCmec type; notably, 17% clinical isolates and 28% community isolates had multiple SCCmec types. Further studies indicated that isolates with multiple SCCmec elements had more stable mecA mRNA expression compared with isolates with single SCCmec elements. CoNS with multiple SCCmec elements demonstrated superior cell wall integrity. Interestingly, phylogenetic analyses of obtained 70 ccr segments indicated that horizontal gene transfer of the ccr complex might exist among various species of clinical CoNS, community CoNS and S. aureus. CONCLUSIONS: CoNS recovered from patients carried extremely diverse but distinctive SCCmec elements compared with isolates from the community. More attention should be given to CoNS with multiple SCCmec not only because they had superior cell wall integrity, but also because CoNS and S. aureus might acquire multiple SCCmec through the ccr complex.


Assuntos
Cromossomos Bacterianos/genética , Coagulase/análise , Resistência a Meticilina/genética , Recombinases/genética , Staphylococcus aureus/genética , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Pequim , China , DNA Bacteriano/genética , Transferência Genética Horizontal , Genes Bacterianos , Humanos , Testes de Sensibilidade Microbiana , Oxacilina/farmacologia , Proteínas de Ligação às Penicilinas/genética , Filogenia , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/isolamento & purificação
18.
Microb Pathog ; 110: 73-77, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28642006

RESUMO

OBJECTIVES: Vulvovaginal candidiasis (VVC) or vaginal candidiasis is a common fungal infection of the genitals causing inflammation, irritation, itching, and vaginal discharge. Common yeast infections are caused by the yeast species C. albicans. However, there are other species of Candida such as C. dubliniensis which are considered as the causative agents of this infection. Hydrolytic enzymes such as proteinase and coagulase are known as virulence factors. The aim of this study was the molecular confirmation and differentiation of C. dubliniensis among C. albicans strains isolated from women with vulvovaginal candidiasis by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) and the evaluation of proteinase and coagulase activities. METHODS: A total of 100 C. albicans strains isolated from women with vulvovaginal candidiasis referred to Shiraz medical clinics were enrolled in the study. All the isolates were primarily identified by conventional methods. PCR-RFLP method was used for the confirmation and identification of C. albicans and C. dubliniensis. Moreover, in vitro proteinase and coagulase activities of these isolates were evaluated using bovine serum albumin media and classical rabbit plasma tube test. RESULTS: As a result, PCR-RFLP identified 100% of the isolates as C. albicans, and no C. dubliniensis could be identified in this study. 84% of the isolates showed proteinase activity, whereas coagulase activity was only detected in 5% of the isolates. CONCLUSIONS: This study reveals that C. dubliniensis plays no role in vaginal candidiasis in Iranian patients. Proteinase production could be an essential virulence factor in C. albicans pathogenicity, but coagulase activity has less potential in this matter.


Assuntos
Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candida/enzimologia , Candida/isolamento & purificação , Candidíase Vulvovaginal/microbiologia , Candida/genética , Candida/patogenicidade , Candida albicans/genética , Candida albicans/patogenicidade , Candidíase Vulvovaginal/diagnóstico , Coagulase/análise , DNA Fúngico/isolamento & purificação , Feminino , Humanos , Irã (Geográfico) , Peptídeo Hidrolases/análise , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição
19.
Int. microbiol ; 20(2): 65-73, jun. 2017. graf, tab
Artigo em Inglês | IBECS | ID: ibc-164431

RESUMO

The present study was conducted to isolate and characterize the molecular epidemiology of the methicillinresistant staphylococci in the general university environment, where all five locations; the library, restrooms, canteens, computer rooms and outdoor surfaces, are in common use by a large population of students. We used Mannitol Salt Agar (MSA) supplemented with 4 µg/ml of oxacillin to screen the methicillin-resistant staphylococci. The species level was identified by PCR of rdr (Staphylococcus epidermidis), groESL (Staphylococcus haemolyticus) and nuc (Staphylococcus aureus and Staphylococcus warneri) genes and DNA sequencing of tuf and dnaJ genes. The susceptibility patterns of the isolates were determined using the disk diffusion method. Antibiotic and disinfectant resistance genes, together with SCCmec types, were detected by the PCR method. The methicillin resistant-staphylococci were isolated from 41 of 200 samples (20.5%), and all of them were found to be methicillin-resistant coagulase negative staphylococci (MR-CoNS). The library had the highest percentage of contamination, with 43.3% of the samples found to be contaminated. All isolates belonged to 6 different species including S. haemolyticus, S. epidermidis, S. warneri, S. cohnii, S. saprophyticus and S. hominis. The antimicrobial resistance rates were highest against penicillin (100%), then cefoxitin (73.1%), erythromycin (73.1%) and oxacillin (68.3%). Altogether, the isolates were approximately 61.0% multidrug resistant (MDR), with the S. epidermidis isolates being the most multidrug resistant (P < 0.05). The prevalence of the qacA/B gene was detected in 63.4% of the isolates, and SCCmec could be typed in 43.9% (18/41) of the isolates. The type range was: II (n = 1), IVd (n = 1), I (n = 2), V (n = 6), IVa (n = 8) and untypeable (n = 23). This result indicates that these university environments are contaminated with methicillin-resistant coagulase negative staphylococci that carry various SCCmec types and high rate of disinfectant resistance genes (AU)


No disponible


Assuntos
Humanos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Infecções Estafilocócicas/epidemiologia , Coagulase/análise , Tailândia/epidemiologia , Resistência Microbiana a Medicamentos , Testes de Sensibilidade Microbiana/métodos
20.
J Hum Lact ; 33(2): 329-340, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28418807

RESUMO

BACKGROUND: Human milk is the preferred nutrition for neonates and a source of bacteria. Research aim: The authors aimed to characterize the molecular epidemiology and genetic content of staphylococci in the human milk of mothers of preterm and term neonates. METHODS: Staphylococci were isolated once per week in the 1st month postpartum from the human milk of mothers of 20 healthy term and 49 preterm neonates hospitalized in the neonatal intensive care unit. Multilocus variable-number tandem-repeats analysis and multilocus sequence typing were used. The presence of the mecA gene, icaA gene of the ica-operon, IS 256, and ACME genetic elements was determined by PCR. RESULTS: The human milk of mothers of preterm compared with term neonates had higher counts of staphylococci but lower species diversity. The human milk of mothers of preterm compared with term neonates more often contained Staphylococcus epidermidis mecA (32.7% vs. 2.6%), icaA (18.8% vs. 6%), IS 256 (7.9% vs. 0.9%), and ACME (15.4% vs. 5.1%), as well as Staphylococcus haemolyticus mecA (90.5% vs. 10%) and IS 256 (61.9% vs. 10%). The overall distribution of multilocus variable-number tandem-repeats analysis (MLVA) types and sequence types was similar between the human milk of mothers of preterm and term neonates, but a few mecA-IS 256-positive MLVA types colonized only mothers of preterm neonates. Maternal hospitalization within 1 month postpartum and the use of an arterial catheter or antibacterial treatment in the neonate increased the odds of harboring mecA-positive staphylococci in human milk. CONCLUSION: Limiting exposure of mothers of preterm neonates to the hospital could prevent human milk colonization with more pathogenic staphylococci.


Assuntos
Recém-Nascido Prematuro/fisiologia , Leite Humano/química , Staphylococcus/isolamento & purificação , Nascimento a Termo/fisiologia , Adulto , Amoxicilina/farmacologia , Amoxicilina/uso terapêutico , Ampicilina/farmacologia , Ampicilina/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Aleitamento Materno , Cefuroxima/farmacologia , Cefuroxima/uso terapêutico , Coagulase/análise , Estônia , Feminino , Humanos , Unidades de Terapia Intensiva Neonatal/organização & administração , Unidades de Terapia Intensiva Neonatal/estatística & dados numéricos , Estudos Longitudinais , Leite Humano/microbiologia , Mães/estatística & dados numéricos , Penicilinas/farmacologia , Penicilinas/uso terapêutico , Estudos Prospectivos , Staphylococcus/metabolismo , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus epidermidis/metabolismo , Staphylococcus haemolyticus/isolamento & purificação , Staphylococcus haemolyticus/metabolismo
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