Results from the Survey of Antibiotic Resistance (SOAR) 2015-17 in Latin America (Argentina, Chile and Costa Rica): data based on CLSI, EUCAST (dose-specific) and pharmacokinetic/pharmacodynamic (PK/PD) breakpoints.

OBJECTIVES: To determine antibiotic susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolates from community-acquired respiratory tract infections (CA-RTIs) collected in 2015-17 from Argentina, Chile and Costa Rica. METHODS: MICs were determined by CLSI broth microdilution and susceptibility was assessed using CLSI, EUCAST (dose-specific) and pharmacokinetic/pharmacodynamic (PK/PD) breakpoints. RESULTS: A total of 170 S. pneumoniae and 218 H. influenzae isolates were collected at five centres in Argentina, Chile and Costa Rica in 2015-17. Small S. pneumoniae isolate numbers from Costa Rica (n = 2) meant that these could only be included in the penicillin susceptibility analysis; they were excluded from further country analyses. Around one-third of pneumococcal isolates from Argentina and two-thirds from Chile were non-susceptible to penicillin by CLSI oral or EUCAST low-dose IV breakpoints, but most (≥89%) were susceptible by CLSI IV or EUCAST high-dose breakpoints. Amongst pneumococci from Argentina, about 80% or more were susceptible to most other antibiotics except cefaclor (all breakpoints), cefixime (PK/PD breakpoints), cefuroxime (EUCAST breakpoints) and trimethoprim/sulfamethoxazole (CLSI and PK/PD breakpoints). S. pneumoniae isolates from Chile showed significantly lower susceptibility (P < 0.05) using CLSI breakpoints compared with those from Argentina for many of the antibiotics tested. Among isolates of H. influenzae from Latin America, more than 90% were susceptible to amoxicillin/clavulanic acid (high dose), cefixime, cefpodoxime, ceftriaxone and fluoroquinolones, irrespective of the breakpoints used. The application of different EUCAST breakpoints for low and higher doses for some of the antibiotics (amoxicillin, amoxicillin/clavulanic acid, ampicillin, penicillin, ceftriaxone, clarithromycin, erythromycin, levofloxacin and trimethoprim/sulfamethoxazole) allowed, for the first time in a SOAR study, the effect of raising the dosage on susceptibility to be quantified. CONCLUSIONS: Antibiotic susceptibility of H. influenzae isolates was generally high in the Latin American countries studied; however, susceptibility profiles varied for S. pneumoniae by country and depending on the breakpoints used, especially for cefaclor. These factors are important in decision making for empirical therapy of bacterial infections.

[In vitro activity of tigecycline in clinical isolates of multidrug resistant Acinetobacter spp. Comparison of different methods of evaluation]. / Actividad in vitro de la tigeciclina en aislamientos clínicos de Acinetobacter spp. multirresistentes. Comparación de diferentes métodos de evaluación.

Infections caused by multidrug resistant (MDRA) Acinetobacter spp. have increased worldwide. Tigecycline provides a new therapeutic option for treating these infections. We conducted a study tailored to validate an in house methodology by broth microdilution (BM) vs. commercial BM in 32 MDRA isolates. Sixty MDRA isolated in 2 time periods (2000-2003 and 2006-2008) were compared by BM, agar dilution (AD) and disk diffusion (DD) methods, as described by the CLSI and the MIC90 for each period was determined. Susceptibility was interpreted by using the breakpoints suggested by the FDA for Enterobacteriaceae. The correlation between the methodologies was performed by a scattergram, and the errors between methods were calculated. The correlation coefficients between AD and BM, and BM and DD were, r: 0.68 in both cases. For 2002-2003, the MIC90 for BM was: 1 g/ml, and for 2006-2008: 2 microg/ml. Using the FDA breakpoints for DD, we observed an unacceptable minor error (17.6%) because of false-intermediate values, considering the MIC90 was 1-2 microg/ml.

[In vitro activity of doripenem and other carbapenems against Pseudomonas aeruginosa]. / Actividad comparativa in vitro de doripenem y de otros carbapenemes frente a Pseudomonas aeruginosa.

Doripenem, a new carbapenem, has shown to be more active against Pseudomonas aeruginosa than other carbapenems. The activity of doripenem, imipenem and meropenem was evaluated against 93 P. aeruginosa isolates, by agar dilution and disk diffusion methods. MIC50 and MIC90, were as follows (microg/ml): doripenem, 2 and 4; meropenem, 2 and 8; and imipenem, 4 and 8, respectively. Doripenem MICs were 1 to 3 dilutions lower (i.e. more active) than those for imipenem in 82% of the isolates. In comparison with meropenem, doripenem was 1 to 3 dilutions more active in 50% of the isolates. Forty-nine percent of isolates showed the same MIC for both antibiotics. Resistance percentages for both methods were (dilution/diffusion): imipenem = 7.5%/49.5% and meropenem = 3.2%/9.7%. As the CLSI has not established cut off values for doripenem yet, resistance rates for this antibiotic were estimated by considering (a) the same cut off values for imipenem/meropenem set up by the CLSI, and (b) those suggested by Brown et al. In case (a), resistance rates would be 1.1%/2.2% whereas in case (b) 1.1%/17.2% for agar dilution and disk diffusion, respectively. In scenarios where resistance to carbapenem is based on mechanisms other than carbapenemases, doripenem has a promising future for treating P. aeruginosa infections.

Uropathogen resistance: are laboratory-generated data reliable enough?

Therapy of uncomplicated urinary tract infection (UTI) is based on knowledge of in vitro susceptibility profiles of uropathogens in the geographic region. Microbiological surveillance systems, which lack epidemiological and clinical data to differentiate between complicated and uncomplicated UTI may incorrectly estimate rates of resistance in the community. We determined the susceptibility profile of bacteria isolated from a random sample of 124 adult outpatients with diagnosis of uncomplicated UTI and we compared it with all outpatient urine specimens collected by the same participant laboratories during the same period. Escherichia coli was the most frequently isolated uropathogen in patients with uncomplicated UTI, and its rate of resistance to different antimicrobials was lower than overall resistance rates to E. coli reported by the participating laboratories during the same period. Resistance to cotrimoxazole was significantly lower. These results suggest that surveillance systems without clinical and epidemiological data may incorrectly gauge uropathogen resistance in the community.

Modified Spot CAMP Test: A rapid, inexpensive and accurate method for identification of group B streptococci.

A rapid modified spot CAMP test using 183 clinical isolates of beta haemolytic streptococci was compared with the standard CAMP test described by Christie et al. The scheme of biochemical identification and serological confirmation was taken as reference method. The sensitivity of both tests was 100%, and the specificity of the rapid and standard tests was 96.8% and 88.9% respectively. The modified spot CAMP test is a rapid, inexpensive and accurate method for the identification of group B streptococci, and is more specific than the standard CAMP test.

[Microbiologic study of bacteremia and fungemia in chronic hemodialysis patients]. / Estudio microbiológico de bacteriemias y fungemias en pacientes en hemodiálisis crónica.

Microbiologic study of bacteremia and fungemia in chronic hemodialysis patients. Bloodstream infections are the second cause of death in patients in chronic hemodialysis (CHD), and the knowledge of the epidemiology is useful to establish proper empiric therapies. The aim of this study was to evaluate the frequency and distribution of microorganisms, in bacteremia and fungemia in 530 patients in CHD. Two hundred and forty eight blood culture series from 114 patients with suspected bacteremia were processed; 44% of them were positive from which 71% (n=78) were clinically significative and belonged to 58 patients. Sixty eight percent of these isolates were gram-positive cocci (n:53), and 22% gram-negative rods (n:17). Staphylococcus aureus was the most prevalent pathogen showing 23% of methicillin-resistance. Candida spp. was the fourth pathogen most common in frequency.

Comparative in vitro bactericidal activity between cefepime and ceftazidime, alone and associated with amikacin, against carbapenem-resistant Pseudomonas aeruginosa strains.

Fifteen unique isolates of carbapenem-resistant Pseudomonas aeruginosa were selected for time-kill studies to assess the bactericidal activity of cefepime (CFP) and ceftazidime (CZD) (at 4 and 16 microg/mL), alone and associated with amikacin (AMK) (4 microg/mL). CFP proved more active than CZD (p < 0.05, Student's t test). Bactericidal activity after 24-h incubation was only achieved by the combination of CFP (16 microg/mL) plus AMK. The higher in vitro activity of cefepime over that of ceftazidime against imipenem-resistant P. aeruginosa strains highlights the differences of these drugs beyond Enterobacterspp. and Staphylococcus aureus.

Comparison of several methods to determine methicillin-resistance in Staphylococcus aureus with focus on borderline strains.

We compared the performance of several phenotypic tests to detect methicillin-resistant Staphylococcus aureus, with special focus on borderline strains. The reliability of the agar screen oxacillin and BBL Crystal tests was asserted for all methicillin-susceptible (n = 25), -resistant (n = 29) and borderline beta-lactamase-hyperproducer (n = 10) strains. Whereas these tests failed to detect 4 of 5 rare borderline strains containing few cells with high-level methicillin resistance (i.e., a frequency of 10(-7)-10(-8)), a "two-temperature" disk diffusion method, performed simultaneously at 35 and 42 degrees C, detected all of such strains.

Activity of gatifloxacin compared to those of seven agents against bacteria recovered from outpatients with respiratory tract infection.

The in vitro activity of gatifloxacin and levofloxacin, ciprofloxacin, penicillin, ampicillin, ampicillin-sulbactam, ceftriaxone and clarithromycin was evaluated against 173 S. pneumoniae strains (128, penicillin-susceptible strains; 32, intermediate penicillin- resistant strains and 13, penicillin-resistant strains), 163 H. influenzae strains (128, beta-lactamase non-producer; 35, beta-lactamase producers), 111 M. catarrhalis (9, beta-lactamase non-producer; 102, beta-lactamase producers), 95 Streptococcus pyogenes and 116 S. aureus strains (96, methicillin-susceptible; 20, methicillin-resistant) recovered from outpatients with respiratory tract infection. Based upon the MICs at which 50% and 90% of the isolates were inhibited we concluded that gatifloxacin proved to be the most active antibiotic against respiratory pathogens, including all the penicillin-resistant pneumococci and H. influenzae or M. catarrhalis producing beta-lactamase. Furthermore, their MICs against S. pneumoniae and methicillin-resistant S. aureus were lower than those of levofloxacin and ciprofloxacin.Therefore, this new fluoroquinolone displayed in vitro features that make it suitable for treating community-acquired respiratory tract infections.

Prospective evaluation of in-house polymerase chain reaction for diagnosis of mycobacterial diseases in patients with HIV infection and lung infiltrates.

SETTING: Rapid diagnosis of tuberculosis (TB) in AIDS is critical for optimal treatment to reduce mycobacterial dissemination, HIV-1 replication and mortality. The inadequate sensitivity of Ziehl-Neelsen staining and its inability to distinguish atypical mycobacteria delays accurate diagnosis. OBJECTIVE: To evaluate the polymerase chain reaction (PCR) for diagnosis of TB in bronchoalveolar lavage (BAL), blood and extra-pulmonary samples from patients with AIDS and pulmonary infiltrates. DESIGN: Specimens from 103 HIV-1-infected patients were prospectively analysed using bacteriological methods and IS6110-PCR. Smear-positive samples were also tested using 16S ribosomal-DNA-PCR to identify Mycobacterium avium complex (MAC) infections. Gold standard diagnosis relied on positive cultures or treatment outcome. RESULTS: Thirty-four patients exhibited TB, one TB and MAC and four MAC. The sensitivity of IS6110-PCR was 100% in smear-positive samples, 81.8% in smear-negative BAL, 66.7% in extra-pulmonary samples and 42.9% in blood. Its specificity was 97.1% in BAL and 100% in extra-pulmonary and blood specimens. The 16S rDNA-PCR identified M. avium from all smear-positive samples that grew MAC. CONCLUSIONS: IS6110-PCR proved useful in evaluating episodes with probable clinical diagnosis of pulmonary or mixed TB and negative smears, whereas 16S rDNA-PCR would be helpful for prompt differential diagnosis of MAC in smear-positive specimens.

Multicentre study of the in vitro evaluation of moxifloxacin and other quinolones against community acquired respiratory pathogens.

The in vitro activity of moxifloxacin was compared with that of ciprofloxacin, levofloxacin, ofloxacin and trovafloxacin against 710 strains (180 Streptococcus pneumoniae, 180 Haemophilus influenzae, 160 Moraxella catarrhalis and 190 Streptococcus pyogenes) isolated from patients with community-acquired respiratory tract infections. MIC values for moxifloxacin, trovafloxacin were 0.25/0.25, 0.03/0.03, 0.06/0.03 and 0.125/0.0125 mg/l for S. pneumoniae, H. influenzae, M. catharralis and S. pyogenes. Based upon the MIC(90) values and the MIC distributions, moxifloxacin and trovafloxacin were the most active of the quinolones tested. They showed enhanced activity against Gram-positive organisms including penicillin non susceptible S. pneumoniae strains. Moxifloxacin was also highly active against ciprofloxacin-resistant S. pneumoniae strains.

Rationale for treating community-acquired lower respiratory tract infections with amoxicillin/sulbactam combination through pharmacodynamic analysis in the setting of aminopenicillin-resistant organisms.

In order to establish a rationale for treating community-acquired lower respiratory tract infections, we assess here the pharmacodynamics of amoxicillin/sulbactam, 500mg/500mg, a formulation marketed in Argentina since 1988 and currently available in 17 countries, against the major pathogens, in comparison with that of a novel formulation (875mg/125mg, see J Chemother 2000; 12: 223-227). In time-kill studies, both bactericidal and inhibitory activity were seen in the 1.5- and 6-h sera, obtained from 12 volunteers after a single oral dose, against both a penicillin-susceptible and an -intermediate Streptococcus pneumoniae strain, as well as against Moraxella catarrhalis and a beta-lactamase-negative Haemophilus influenzae strain. Only the 1.5-h sera proved bactericidal against a penicillin-resistant S. pneumoniae strain (MIC, 2 microg/ml) and a beta-lactamse-positive H. influenzae isolate. This study suggests that amoxicillin/sulbactam (500mg/500mg) is still a suitable option for treating community-acquired lower respiratory tract infections, allowing a b.i.d. dosing schedule. Caution should be taken with pneumonia caused by beta-lactamase-positive H. influenzae or penicillin-resistant (MIC > or =2 microg/ml) S. pneumoniae isolates. Either shorter dosing intervals (t.i.d.) or a higher amoxicillin content in the formulation (i.e. 875 mg) may be required in these situations.

A pharmacodynamic model to support a 12-hour dosing interval for amoxicillin/sulbactam, a novel oral combination, in the treatment of community-acquired lower respiratory tract infections.

We evaluated, by time-kill studies, the pharmacodynamics of amoxicillin/sulbactam (AMX/SUL, 875 mg/125 mg), a novel oral combination, against the major respiratory pathogens in 12 volunteers receiving a single dose. The sera corresponding to 50% of a 12-h dosing interval displayed either bactericidal or inhibitory activity against both a penicillin-susceptible and a penicillin-intermediate Streptococcus pneumoniae strain (penicillin MIC of 0.03 and 0.25 microg/ml, respectively), as well as against a beta-lactamase-positive Moraxella catarrhalis and a beta-lactamase-negative Haemophilus influenzae strain. Both the peak samples and those corresponding to 4 h after dose (i.e. 33% of a 12-h dosing interval) proved active against both a penicillin-resistant S. pneumoniae (MIC, 2 microg/ml) and a beta-lactamase-positive H. influenzae strain. The AMX-SUL formulation evaluated in this study showed pharmacodynamic features that support clinical trials to assess its efficacy in the treatment of lower respiratory tract infections with a 12-h dosing interval regimen.

[Endocarditis due to Corynebacterium xerosis]. / Endocarditis por Corynebacterium xerosis.

Corynebacterium sp. are found as normal flora in skin and mucosal sites. They have been isolated in empyemas, brain abscesses, blood cultures and ventricular shunts. About 9-10% of early-onset and 4-5% late-onset prosthetic valve endocarditis are due to different species of the so-called "diphteroids". A 30 year-old white female was admitted after 30 days with fever of undetermined origin. A mitral prosthesis had been fitted in 1977. On physical examination a protomesosystolic mitral murmur, petechiae, retinal hemorrhages and hepatosplenomegaly were detected. Laboratory tests showed 37% hematocrit, 14,800/mm3 white blood cells, 78 mm ESR, urinary sediment: less than 30/h.p.f. red blood cells. A new first-degree A-V block was detected. Blood cultures were negative. Due to persistent fever, progressive anemia, leukocytosis and new vegetations on echocardiogram, surgery was performed. A mitral valve ring abscess was found. Corynebacterium xerosis was isolated from surgical specimens. The strain was found susceptible to penicillin, ampicillin, oxacillin, ticarcillin, piperacillin, cephalotin, cefoxitin, cefoperazone, rifampin, gentamicin, amikacin, and norfloxacin. Studies with clindamycin, disclosed MIC and MBC = 0.25 mg/l. The patient received 1800 mg/day clindamycin for 4 weeks. Serum cidal studies showed a peak concentration 1/128 and a titre of trough 1/4. Negative control blood cultures were obtained. She has remained well for nine months after treatment. Corynebacterium sp. can cause "apparently" negative blood cultures. Blood samples should be incubated for more than 15 days before they can be considered negative. Almost 50% of previously described cases have been detected during the six months after cardiac surgery. Mortality has been high (48%).(ABSTRACT TRUNCATED AT 250 WORDS)

[Specificity against group A, C, and G streptococci of the reverse CAMP test for identifying Clostridium perfringens]. / Especificidad frente a estreptococos grupo A, C y G de la prueba de "CAMP reversa" en identificación de Clostridium perfringens.

We studied "reverse CAMP" test specificity for the identification of Clostridium perfringens, and the probable existence of cross-reactions with groups A, C and G streptococci. Ninety eight Clostridium strains were tested with ten S. agalactiae, ten S. pyogenes, two group C streptococci and one group G. All the strains of S. agalactiae yielded a positive "reverse CAMP" reaction when it was performed with C. perfringens strains. Cross-reactions were not detected when C. perfringens were tested with groups A, C or G streptococci. The "reverse CAMP" test proved to be specific for all the C. perfringens strains used.

[Non-pigmented Serratia: utility of hydrolysis of Tween 80 in its identification]. / Serratia no pigmentadas: utilidad de la hidrólisis del Tween 80 en su identificación.

The ability of Serratia sp. to hydrolyze Tween 80 and the usefulness of this test for differentiating it from other Gram negative bacilli was studied, comparing it with the capacity of production of DNAsa by the same strains using the O-Toluidine blue method. The total number of strains assayed was 88. The sensitivity obtained was 83.9% and specificity 91.3%, with a positive predictive value of 83.9% and a negative predictive value of 91.2%. Using this method to differentiate only non-pigmented Serratia from Klebsiella sp. and Enterobacter sp. the sensitivity and specificity obtained was 100%. Concerning the cost, this method proved to be 3.86 times cheaper than the DNAsa one.

[Utility of pyrrolidonyl-arylamidase detection for typing Enterobacteriaceae and non-fermenting Gram-negative bacteria]. / Utilidad de la detección de pirrolidonil-arilamidasa en la diferenciación de enterobacterias y bacilos gram-negativos no fermentadores.

Detection of pyrrolidonyl-aryl-amidase activity (PYR) is an important tool to identify gram-positive cocci, such as staphylococci, enterococci, streptococci, and other related genera. However, only few studies evaluating its usefulness with gram-negative rods have been published. Thus, a prospective study including 542 and 215 unique clinical isolates of Enterobacteriaceae and non-fermentative gram-negative rods, respectively, was undertaken. Strains were identified by conventional methods. PYR test was performed using a commercial kit, according to the manufacturer recommendations. Positive results were uniformly obtained for the PYR test with the following species: Citrobacter spp, Klebsiella spp, Enterobacter aerogenes, Enterobacter agglomerans group, Serratia marcescens and S. odorifera. On the other hand, negative results were uniformly displayed by E. coli (including inactive E. coli), Protease group, Salmonellia spp, Shigella spp, Acinetobacter spp, Burkholderia (Pseudomonas) cepacia and Flavobacterium spp. Variable results were shown in Pseudomonas aeruginosa, Stenotrophomonas (xanthomonas) malthophilia, Kluyvera cryocrescens, and Enterobacter cloacae. PYR test proved to be a reliable and simple tool to rapidly distinguish certain species belonging to Enterobacteriaceae (ie. Citrobacter freundii from Salmonella spp, and inactive E. coli from K. ozaenae). Further studies, including a wide diversity of species, are required to assess usefulness of the PYR test for the identification of non-fermentative gram-negative rods.

[Infection of a cerebrospinal fluid shunt system by Bacillus circulans and Bacillus larvae]. / Infección del sistema de derivación del LCR por Bacillus circulans y Bacillus larvae.

A two episodes case of CSF ventriculo-atrial shunt infection due to B. circulans and B. larvae is presented. B. circulans was first isolated from 4 blood cultures and CSF (shunt valve tap). The patient showed a brain damage syndrome reversible with antibiotic treatment. Lethal toxin production was demonstrated for the B. circulans strain in a mouse model. This strain was found to be a variant of Gordon's description as it produced urease and was tolerant to 7% NaCl. The patient recovered after cefotaxime, cotrimoxazole and rifampicin treatment. A second infection due to B. larvae was detected two months later. The shunt system was removed due to obstruction and a scanning electron microscopy study was performed. Confluent masses of white blood cells and rods were observed on the inner surface of the catheter. As for as we know, this is the first case of human infection due to B. larvae.

[Clostridium difficile diarrhea: frequency of detection in a medical center in Buenos Aires, Argentina]. / Diarrea asociada a Clostridium difficile: frecuencia de detección en un centro médico de Buenos Aires, Argentina.

Clostridium difficile has been recognized as the most important enteric pathogen of nosocomial antibiotic-associated diarrhea (CDAD) in adults from industrialized countries. The importance of C. difficile as a cause of diarrhea in ambulatory patients appears underestimated or under-recognized. Since the 1980's, outbreaks of CDAD have been increasingly reported, but there are few data available in Argentina. We developed a retrospective study to provide some information about CDAD in our country. From July 1998 to November 1999, a total of 245 fecal specimens from hospitalized and some ambulatory patients were tested in order to confirm the diagnosis of CDAD. C. difficile cytotoxin (toxin B) was identified by detecting its cytopathic effect on monolayers of McCoy culture cells. For culture and isolation of C. difficile, stool samples were prepared by ethanol shock prior to plating onto a selective medium which contained blood, cefoxitin and fructose. Of the 245 samples, 14 (5.8%) were identified as positive by the cell cytotoxicity assay. Using the criteria of isolation of cytotoxigenic C. difficile positivity increased to 6.5% (16 samples). Thirteen of the positive results were from hospitalized patients (81.3%) and 3 (18.7%) from outpatients. The mean age of inpatients was 72.9 years (ranging from 47 to 88). All patients had received 2 or more antimicrobial agents (most of them beta-lactams) 2 months before the appearance of diarrhea. There was one patient who had received only chemotherapy. The prevalence of CDAD in this study was less than in others previously reported. This difference may be due to the fact that not all general practitioners include testing for C. difficile when the patient with diarrhea had previously received antibiotics. More educational programs should be directed to all physicians, concerning the role of C. difficile as an important enteric pathogen in patients who have undergone treatment with antimicrobial or chemotherapeutic agents.