CARACTERIZAÇÃO DO STAPHYLOCOCCUS COAGULASE NEGATIVO ISOLADOS DE QUARTOS MAMÁRIOS DE VACAS COM MASTITE SUBCLÍNICA

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pesquisa dos genes icaD e bap de adesão intercelular bacteriana e formação de biofilme por Staphylococcus aureus e Staphylococcus coagulase-negativa isolados de mastite bovina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Perfil microbilógico de diferentes tipos de saladas provenientes de cozinhas hospitalares

Pós-graduação em Medicina Veterinária - FMVZ

Características genotípicas de Staphylococcus coagulase-negativos e taxas de cura da mastite ovina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Assessment of the risk of raw milk consumption related to staphylococcal food poisoning

This study aimed to identify the risks of staphylococcal food poisoning due to the consumption of raw milk. Fifty-one farms in Londrina (PR) and 50 in Pelotas (RS) were analyzed, to determine the population of coagulase-positive staphylococci (UFC/ mL), as well as to verify the ability of producing Staphylococcal Enterotoxin A (SEA) by immunodifusion (OSP), the presence of the gene for the production of SEA (PCR) in the cultures, and the research of enterotoxin (SEA to SEE) in milk samples using ELISA commercial kit. Considering the 101 farms analyzed, 19 (18.8%) presented coagulase-positive staphylococci count above 105 UFC/mL. For the evaluation of the enterotoxigenic ability (SEA) by the OSP technique, six cultures coagulase-positive (5.5%) were positive to the test and identified as S. aureus. From the coagualse-negative sample, one (5.5%) was OSP positive. For the evaluation of the presence of the gene for EEA synthesis, 51 cultures of staphylococci were tested. From this total, 14 (27.45%) presented the gene, and from that, only 5 (9.81%) cultures were capable of expressing it in the technique of the OSP. The morphologic characteristic of the evaluated cultures that had enterotoxigenic capacity, from the 14 (33,3%) cultures that presented the gene for EEA production, 05 (11.9%) were characterized as typical cultures of S.aureus in Baird Parker agar. All the 12 milk samples studied for the presence of EEA to EEE in milk were negative. Thus, it can be concluded that there is extensive contamination of raw milk for staphylococci coagulase, however, most of the isolated strains were not enterotoxigenic or did not express such a characteristic. Only 9.81% of the tested colonies expressed the gene and effectively produced SEA. None of the samples had sufficient counts to produce detectable amounts of SEA. The milk samples did not present risk to cause staphylococcal food poisoning if consumed in natura until the collection moment.

Low rate of methicillin-resistant coagulase-positive staphylococcal colonization of veterinary personnel in Hong Kong

Elevated rates of methicillin-resistant Staphylococcus aureus (MRSA) carriage have been reported in veterinary personnel, suggesting an occupational colonization risk. Hong Kong veterinary personnel (n = 150) were sampled for coagulase-positive staphylococci (CPS) nasal colonization. Risk factors for colonization were assessed by questionnaire. Isolates were identified and antibiotic susceptibility determined. All CPS isolates were investigated for mecA carriage, SCCmec type and PVL genes. Two subjects were colonized with methicillin-resistant CPS: one with MRSA (spa type t002 (CC5), SCCmec type II) and one with methicillin-resistant Staphylococcus pseudintermedius (MRSP) (MLST type ST71, SCCmec type II-III). MLST type ST71 S. pseudintermedius strain is the predominant MRSP clone circulating in dogs in Europe and in Hong Kong. The low MR-CPS colonization rate may be associated with low levels of large animal exposure or low rates of MRSA colonization of companion animals in Hong Kong. Colonization with non-aureus CPS, which may cause human infection, must also be considered in veterinary personnel.

Presence of new mecA and mph(C) variants conferring antibiotic resistance in Staphylococcus spp. isolated from the skin of horses before and after clinic admission

Because of the frequency of multiple antibiotic resistance, Staphylococcus species often represent a challenge in incisional infections of horses undergoing colic surgery. To investigate the evolution of antibiotic resistance patterns before and after preventative peri- and postoperative penicillin treatment, staphylococci were isolated from skin and wound samples at different times during hospitalization. Most staphylococci were normal skin commensals and belonged to the common coagulase-negative group. In some cases they turned out to be opportunistic pathogens present in wound infections. MICs were determined for 12 antibiotics, and antibiotic resistance genes were detected by microarray. At hospital admission, horses harbored staphylococci that were susceptible to antibiotics or resistant to one group of drugs, mainly due to the presence of new variants of the methicillin and macrolide resistance genes mecA and mph(C), respectively. After 3 days, the percentage of Staphylococcus isolates displaying antibiotic resistance, as well as the number of resistance genes per isolate, increased moderately in hospitalized horses without surgery or penicillin treatment but dramatically in hospitalized horses after colic surgery as well as penicillin treatment. Staphylococcus species displaying multiple resistance were found to harbor mainly genes conferring resistance to beta-lactams (mecA and blaZ), aminoglycosides [str and aac(6')-Ie-aph(2')-Ia], and trimethoprim [dfr(A) and dfr(D)]. Additional genes conferring resistance to macrolides [mph(C), erm(C), and erm(B)], tetracycline [tet(K) and tet(M)], chloramphenicol [cat(pC221) and cat(pC223)], and streptothricin (sat4) appeared in several strains. Hospitalization and preventive penicillin use were shown to act as selection agents for multidrug-resistant commensal staphylococcal flora.

Strain typing and antimicrobial susceptibility of methicillin-resistant coagulase-positive staphylococcal species in dogs and people associated with dogs in Thailand

AIMS This study was to investigate and to characterize methicillin-resistant coagulase-positive staphylococci (MRCoPS) harboring in dogs and people associated with dogs in Thailand. METHODS AND RESULTS Staphylococci were collected from 100 dogs, 100 dog owners, 200 small animal veterinarians and 100 people without pet association. Species of MRCoPS were identified phenotypically and genotypically. Molecular characteristics were determined by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and SCCmec typing, and antimicrobial susceptibility was assayed by broth microdilution and by microarray analysis for resistance genes. Methicillin-resistant Staphylococcus pseudintermedius (MRSP), methicillin-resistant Staphylococcus schleiferi subsp. coagulans (MRSSc) and methicillin-resistant Staphylococcus aureus (MRSA) were isolated from dogs (45, 17 and 1%, respectively), veterinarians (8, 2 and 1·5%, respectively) and dog owners (3, 2 and 0%, respectively). Seventeen sequence types (STs) were identified among 83 MRSP isolates which specifically carried SCCmec V, II-III, ΨSCCmec57395 and three uncharacterized SCCmec types. MRSP ST 45, 68 and novel STs including 169, 178, 181 and 183 were shared among canine and human isolates. Most of MRSA ST398 and MRSSc carried SCCmec type V. The MRCoPS commonly displayed multiple resistances to tested antimicrobials and carried various resistance genes. CONCLUSION Variety of MRCoPS, especially new MRSP clones, distributed in dogs and people in Thailand. SIGNIFICANCE AND IMPACT OF THE STUDY The existence of MRCoPS circulating between dogs and humans in Thailand provides indirect evidence of interspecies transmission and represents a potential public health hazard.

Rapid serodiagnosis of Staphylococcus aureus surgical site infection following median sternotomy

Objectives: To determine the sensitivity and specificity of a novel ELISA for the serodiagnosis of surgical site infection (SSI) due to staphylococci following median sternotomy. Methods: Twelve patients with a superficial sternal SSI and 19 with a deep sternal SSI due to Staphylococcus aureus were compared with 37 control patients who also underwent median sternotomy for cardiac surgery but exhibited no microbiological or clinical symptoms of infection. A further five patients with sternal SSI due to coagulase-negative (CoNS) staphylococci were studied. An ELISA incorporating a recently recognised exocellular short chain form of lipoteichoic acid (lipid S) recovered from CoNS, was used to determine serum levels of anti-lipid S IgG in all patient groups. Results: Serum anti-lipid S IgG titres of patients with sternal SSI due to S. aureus were significantly higher than the control patients (P<0.0001). In addition, patients with deep sternal SSI had significantly higher serum anti-lipid S IgG titres than patients with superficial sternal SSI (P=0.03). Serum anti-lipid S IgG titres of patients with sternal SSI due to CoNS were significantly higher than the control patients (P=0.001). Conclusion: The lipid S ELISA may facilitate the diagnosis of sternal SSI due to S. aureus and could also be of value with infection due to CoNS. © 2005 Published by Elsevier Ltd. on behalf of The Bristish Infection Society.

Microbiologia dos seios maxilar e etmoidal em pacientes com rinossinusite crônica submetidos à cirurgia funcional endoscópica dos seios paranasais

Estudos da microbiologia da rinossinusite crônica mostram a presença de microorganismos aeróbicos, anaeróbicos, fungos e vírus e sua incidência varia de acordo com cada estudo. Estes estudos nos guiam para a escolha do antimicrobiano mais adequado para eliminar o processo infeccioso, ajudando a restaurar a mucosa nasossinusal. FORMA DE ESTUDO: Clínico prospectivo. OBJETIVO: O objetivo deste trabalho foi estudar a microbiologia dos seios maxilar e/ou etmoidal de pacientes com rinossinusite crônica e com indicação de cirurgia funcional endoscópica dos seios paranasais. MATERIAIS E MÉTODOS: Durante a cirurgia coletamos, em 41 pacientes, secreção e/ou fragmento de mucosa dos seios maxilar e/ou etmoidal para realização de bacterioscopia, pesquisa direta de fungos, cultura para microorganismos aeróbios, anaeróbios e fungos. RESULTADOS: Identificou-se a presença de microorganismos aeróbios em 21 pacientes (51,2%), anaeróbios em 16 (39%) e fungos em 1 (2,4%). Na população estudada, apenas em 12 (29,2%) o microorganismo isolado foi considerado patogênico quando analisado junto à contagem semiquantitativa de leucócitos. O Staphylococcus coagulase-negativo e o Staphylococcus aureus foram os microorganismos mais freqüentes, em 5 (12,1%) e em 4 pacientes (9,75%) respectivamente. CONCLUSÃO: Este estudo revela que o Staphylococcus coagulase-negative e o Staphylococcus aureus foram os microorganismos mais freqüentes isolados nos pacientes com rinossinusite crônica.

Sepsis in burned patients

A prospective study was conducted from June 2001 to May 2002 at the Burns Unit of Hospital Regional da Asa Norte, Brasília, Brazil. During the period of the study, 252 patients were treated at the Burns Unit, 49 (19.4%) developed clinically and microbiologically proven sepsis. Twenty-six (53.1%) were males and 23 (46.9%) females with a mean age of 22 years (range one to 89 years) and mean burned body surface area of 37.7 ± 18.4% (range 7 to 84%). Forty-three patients had flame burns, five a scald and one an electric burn. These 49 patients had a total of 62 septic episodes. Forty (81.6%) patients had only one and nine (18.4%) had up to three episodes of sepsis. Thirty (61.2%) patients had their first septicemic episode either earlier or by one week postburn. Out of 62 septic episodes, 58 were due to bacteria and four due to Candida sp. The most common bacteria isolated from blood culture were Staphylococcus aureus, coagulase-negative Staphylococcus, Acinetobacter baumannii, Enterobacter cloacae and Klebsiella pneumoniae. Eleven (18.9%) episodes were due to oxacillin resistant Staphylococcus aureus. Acinetobacter baumannii was sensitive to ampicillin/sulbactam in 71.4% and to imipenem in 85.7% of the cases. The primary foci of sepsis were the burn wound in 15 ( 24.2% ) episodes. The most common clinical findings of sepsis in these patients were fever, dyspnea, hypotension and oliguria. The most common laboratory findings of these patients were anemia, leukocytosis, hypoalbuminemia and thrombocytopenia. Twelve (24.5%) patients died. The appropriate knowledge of clinical, epidemiological, laboratorial and microbiological aspects of sepsis in burned patients permits an adequate diagnosis and treatment of this complication.

Nosocomial infections in a Brazilian neonatal intensive care unit: a 4-year surveillance study

INTRODUCTION: Report the incidence of nosocomial infections, causative microorganisms, risk factors associated with and antimicrobial susceptibility pattern in the NICU of the Uberlândia University Hospital. METHODS: Data were collected through the National Healthcare Safety Network surveillance from January 2006 to December 2009. The patients were followed five times/week from their birth to their discharge or death. RESULTS: The study included 1,443 patients, 209 of these developed NIs, totaling 293 NI episodes, principally bloodstream infections (203; 69.3%) and conjunctivitis (52; 17.7%). Device-associated infection rates were as follows: 17.3 primary bloodstream infections per 1,000 central line-days and 3.2 pneumonias per 1000 ventilator-days. The mortality rate in neonates with NI was 11.9%. Mechanical ventilation, total parenteral nutrition, orogastric tube, previous antibiotic therapy, use of CVC and birth weight of 751-1,000g appeared to be associated with a significantly higher risk of NI (p < 0.05). In multiple logistic regression analysis for NI, mechanical ventilation and the use of CVC were independent risk factors (p < 0.05). Coagulase- negative Staphylococcus (CoNS) (36.5%) and Staphylococcus aureus (23.6%) were the most common etiologic agents isolated from cultures. The incidences of oxacillin-resistant CoNS and S. aureus were 81.8% and 25.3%, respectively. CONCLUSIONS: Frequent surveillance was very important to evaluate the association of these well-known risk factors with NIs and causative organisms, assisting in drawing the attention of health care professionals to this potent cause of morbidity.

Urinary tract infection in full-term newborn infants: value of urine culture by bag specimen collection

OBJETIVE: to evaluate the efficacy of urine culture by bag specimen for the detection of neonatal urinary tract infection in full-term newborn infants. Retrospective study (1997) including full-term newborn infants having a positive urine culture (>100,000 CFU/ml) by bag specimen collection. The urinary tract infection diagnosis was confirmed by positive urine culture (suprapubic bladder aspiration method). The select cases were divided into three groups, according to newborn infant age at the bag specimen collection: GI (< 48 h, n = 17), GII (48 h to 7 d, n = 35) and GIII (> 7 d, n = 9). Sixty one full-term newborn infants were studied (5.1 % of total infants). The diagnosis was confirmed on 19/61 (31.1 %) of full-term infants born alive. Distribution among the groups was: GI = 2/17 (11.8 %), GII = 10//35 (28.6 %), and GIII = 7/9 (77.7 %). The most relevant clinical symptoms were: fever (GI - 100 %, GII - 91.4 %) and weight loss (GI - 35.3 %, GII - 45.7 %). Urine culture results for specimens collected by suprapubic aspiration were: E. coli GI (100 %), GII (40 %) and GIII (28.6 %), E. faecalis GI (30%), Staphylococcus coagulase-negative GII (20 %) and GIII (42.8 %), and Staphylococcus aureus GII (10 %). Correlation between positive urine culture collection (bag specimen method) and urinary tract infection diagnosis, using relative risk analysis, produced the following results: GI=0.30 (CI95% 0.08-1.15), GII=0.51 (CI 95% 0.25-1.06) and GIII=3.31 (CI95% 1.8-6.06) The most frequent urinary tract infection clinical signs in the first week were fever and weight loss, while non-specific symptomatology occurred later. E. coli was most frequent infectious agent, although from the 7th day of life, staphylococcus was noted. The urine culture (bag specimen method) was effective in detecting urinary tract infection only after the 7th day of life.

Clinical presentation, etiology, and outcome of infective endocarditis in the 21st century: the International Collaboration on Endocarditis-Prospective Cohort Study.

We sought to provide a contemporary picture of the presentation, etiology, and outcome of infective endocarditis (IE) in a large patient cohort from multiple locations worldwide. Prospective cohort study of 2781 adults with definite IE who were admitted to 58 hospitals in 25 countries from June 1, 2000, through September 1, 2005. The median age of the cohort was 57.9 (interquartile range, 43.2-71.8) years, and 72.1% had native valve IE. Most patients (77.0%) presented early in the disease (<30 days) with few of the classic clinical hallmarks of IE. Recent health care exposure was found in one-quarter of patients. Staphylococcus aureus was the most common pathogen (31.2%). The mitral (41.1%) and aortic (37.6%) valves were infected most commonly. The following complications were common: stroke (16.9%), embolization other than stroke (22.6%), heart failure (32.3%), and intracardiac abscess (14.4%). Surgical therapy was common (48.2%), and in-hospital mortality remained high (17.7%). Prosthetic valve involvement (odds ratio, 1.47; 95% confidence interval, 1.13-1.90), increasing age (1.30; 1.17-1.46 per 10-year interval), pulmonary edema (1.79; 1.39-2.30), S aureus infection (1.54; 1.14-2.08), coagulase-negative staphylococcal infection (1.50; 1.07-2.10), mitral valve vegetation (1.34; 1.06-1.68), and paravalvular complications (2.25; 1.64-3.09) were associated with an increased risk of in-hospital death, whereas viridans streptococcal infection (0.52; 0.33-0.81) and surgery (0.61; 0.44-0.83) were associated with a decreased risk. In the early 21st century, IE is more often an acute disease, characterized by a high rate of S aureus infection. Mortality remains relatively high.

Determination of bacterial quantity by sonication in vacum-assisted closure (VAC) foams used for treatment of chronic wounds

Background: Negative pressure wound treatment is increasingly used through a Vacuum-Assisted Closure (VAC) device in complex wound situations. For this purpose, sterile polyurethane (PU) and polyvinyl alcohol (PVA) foam dressings are fitted to the wound size and covered with an adhesive drape to create an airtight seal. Little information exists about the type and quantity of microorganisms within the foams. Therefore, we investigated VAC foams after removal from the wound using a validated method (sonication) to detect the bacterial bioburden in the foam consisting as microbial biofilms.Methods: We prospectively included VAC foams (PU and PVA, KCI, Rümlamg, Switzerland) without antibacterial additions (e.g. silver), which were removed from wounds in patients with chronic ulcers from January 2007 through December 2008. Excluded were patients with acute wound infection, necrotizing fasciitis, underlying osteomyelitis or implant. Removed foams from regular changes of dressing were aseptically placed in a container with 100 ml sterile Ringer's solution. Within 4 hours after removal, foams were sonicated for 5 min at 40 kHz (as described in NEJM 2007;357:654). The resulting sonication fluid was cultured at 37°C on aerobic blood agar plates for 5 days. Microbes were quantified as No. of colony-forming units (CFU)/ml sonication fluid and identified to the species level.Results: A total of 68 foams (38 PU and 30 PVA) from 55 patients were included in the study (median age 71 years; range 33-88 years, 57% were man). Foams were removed from the following anatomic sites: sacrum (n=29), ischium (n=18), heel (n=13), calves (n=6) and ankle (n=2). The median duration of being in place was 3 days (range, 1-8 days). In all 68 foams, bacteria were found in large quantities (median 105 CFU/ml, range 102-7 CFU/ml sonication fluid. No differences were found between PU and PVA foams. One type of organisms was found in 11 (16%), two in 17 (24%) and 3 or more in 40 (60%) foams. Gram-negative rods (Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa) were isolated in 70%, followed by Staphylococcus aureus (20%), koagulase-negative staphylococci, streptococci (8%), and enterococci (2%).Conclusion: With sonication, a high density of bacteria present in VAC foams was demonstrated after a median of 3 days. Future studies are needed to investigate whether antimicrobial-impregnated foams can reduce the bacterial load in foams and potentially improve wound healing.