Clostridium ramosum rapidly identified by MALDI-TOF MS. A rare gram-variable agent of bacteraemia.

Clostridium ramosum is an enteric anaerobic, endospore-forming, gram-positive rod with a low GC content that is rarely associated with disease in humans. We present a case of C. ramosum bacteraemia. To the best of our knowledge, this is the second case of C. ramosum bacteraemia in an elderly patient presenting with fever, abdominal pain and bilious emesis. We highlight the Gram stain variability, the lack of visualization of spores and the atypical morphology of the colonies that showed C. ramosum in a polymicrobial presentation that initially appeared to show monomicrobial bacteraemia. The microorganism was rapidly identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We present a comprehensive literature review of 32 cases of clinical infections by C. ramosum in which we describe, if available, sex, age, clinical symptoms, predisposing conditions, other organisms present in the blood culture, other samples with C. ramosum , identification methodology, treatment and outcome.

Unusual presentations of Comamonas kerstersii infection.

The association of Comamonas kerstersii with peritonitis resulting from the presence of perforated appendix has previously been described by our research team. In the present study, we describe the isolation of this microorganism from two forms of unusual presentations of C. kerstersii infection not previously described in the literature: localized intra-abdominal infection (psoas abscess) and pelvic peritonitis.

Survey of Aspergillus section Flavi presence in agricultural soils and effect of glyphosate on nontoxigenic A. flavus growth on soil-based medium.

AIMS: To evaluate the cultivable mycobiota from agricultural soils exposed to pesticides, the aflatoxigenic capacity of Aspergillus section Flavi strains and the effect of glyphosate on lag phase and growth rates of native nontoxigenic Aspergillus flavus under different water potential (MPa) conditions on soil-based medium. METHODS AND RESULTS: Culturable mycobiota analysis from different agricultural soils was performed by the surface spread method. The effect of glyphosate (0-20 mmol l(-1)) on the growth of A. flavus strains was evaluated on a soil extract solid medium. Mycobiota analysis of crop soils showed the presence of twenty-one genera of filamentous fungi. Aspergillus flavus and Aspergillus niger aggregate strains were isolated from the three soil types. Ninety-two per cent of A. flavus strains were toxigenic. In vitro assay results showed that at -0·70 MPa, a significant increase in growth rate in all strains was recorded at 5 and 20 mmol l(-1) of glyphosate. At -2·78 MPa, this parameter remained constant at all glyphosate concentrations, except in GM4 strain where an increase in growth rate was recorded with increasing pesticide concentrations. At -7·06 MPa, a significant increase in growth rate has also been observed in GM 3 strain with 5 mmol l(-1) and in GM 4 strain with 10 and 20 mmol l(-1). CONCLUSIONS: This study showed that the imperfecti fungi Aspergillus spp., Penicillium spp., Trichoderma spp., Cladosporium spp. and Paecilomyces spp. are isolated as prevalent groups in agricultural soil exposed to pesticides, and the capacity of nontoxigenic A. flavus strains to tolerate different glyphosate concentrations under different water potential (MPa) conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This manuscript makes a contribution to the knowledge of cultivable fungal populations from agricultural soils exposed to pesticides and the glyphosate tolerance of A. flavus strains.

Survey of Aspergillus section Flavi and aflatoxin B(1) in brewer's grain used as pig feedstuff in Córdoba, Argentina.

Brewing industry by-products are important animal feedstuff alternatives for local swine producers in Córdoba, Argentina. The high content of nutrients makes these by-products vulnerable to bacterial and fungal contamination. The objectives of the present study were (1) to determine the presence of Aspergillus section Flavi in brewer's grain used to feed pigs and (2) to evaluate the incidence of aflatoxin B(1) in the substrate. Total fungal count of most samples exceeded the levels proposed as feed quality limits, and most Aspergillus section Flavi strains found were able to produce high amounts of AFB(1) in vitro. However, the incidence of AFB(1) was low. The presence of contamination by aflatoxicogenic species in feedstuff might affect the productivity of swine producers and indirectly represents a public health issue.

Food-grade antioxidants and antimicrobials to control growth and ochratoxin a production by Aspergillus section Nigri on peanut kernels.

Each year, a significant portion of the peanuts produced cannot be marketed because of fungal disease at the postharvest stage and mycotoxin contamination. Antioxidants could be used as an alternative to fungicides to control ochratoxigenic fungi in peanuts during storage. This study was carried out to determine the effect of the antioxidant butylated hydroxyanisole (BHA) and the antimicrobial propyl paraben (PP) on the lag phase before growth, growth rate, and ochratoxin A (OTA) production by Aspergillus section Nigri strains in peanut kernels under different conditions of water activity (aw) and temperature. At 20 mM/g BHA, 18 degrees C, and 0.93 aw, complete inhibition of growth occurred. For PP, there was no growth at 20 mM/g, 18 degrees C, and 0.93, 0.95, and 0.98 aw. BHA at 20 mM/g inhibited OTA production in peanuts by Aspergillus carbonarius and Aspergillus niger aggregate strains at 0.93 aw and 18 degrees C. PP at 20 mM/g completely inhibited OTA production at 18 degrees C. The results of this work suggest that PP is more appropriate than BHA for controlling growth and OTA production by Aspergillus section Nigri species in peanut kernels.

Ecophysiological factor effect on growth rate, lag phase and ochratoxin A production by Aspergillus niger aggregate strains on irradiated peanut seeds.

The aims of this study were to evaluate the effects of water activity, temperature, incubation time and their interactions on lag phase, growth rate and ochratoxin A (OTA) production by strains belonging to the Aspergillus niger aggregate on irradiated peanut seeds. In the temperature and water activity range assayed, the optimal conditions of growth for RCP42 and RCP176 strains on irradiated peanut seeds were 0.995 a(w) and 30 degrees C being the growth rates of 12.4 and 14.6 mm/day, respectively. The maximum OTA production occurred at 0.973 a(w) and 25 degrees C for both strains assayed; whereas the minimum OTA production was obtained at 0.951 a(w) and 15 degrees C, at 14 and 21 days of incubation for RCP42 and RCP176 strains, respectively. The amount of OTA accumulated during 21 days assayed by both strains varied from 6.5 to 460 microg/g and from 10 to 210 microg/g with mean levels of 119.2 and 97.5 microg/g for RCP42 and RCP176 strains, respectively. The variance analysis (ANOVA) revealed that OTA concentration produced by RCP42 strain was significantly (p<0.0001) greater than that produced by RCP176 strain. If the strains with which the experiments were carried out were representative of the Aspergillus niger aggregate toxigenic species and the water activity in peanut seeds stored at 0.910 or lower was maintained, OTA production would be reduced during at least 21 days at variable temperatures.

Ochratoxin A production in relation to ecophysiological factors by Aspergillus section Nigri strains isolated from different substrates in Argentina.

Contamination of foodstuff with mycotoxins such as ochratoxins is a major matter of concern for human and animal health. In Aspergillus species, ochratoxin synthesis depends on several environmental factors. The aims of this work were to evaluate the effect of water activity (0.995-0.85), temperature (15, 25 and 30 degrees C), incubation time (7, 14 and 21 days) and their interactions on OTA production on peanut, maize kernels, dried grapes and coffee beans meal extract agar medium by eight strains of Aspergillus section Nigri isolated from human food in Argentina. The optimum temperature for OTA production was 25 or 30 degrees C depending on the strains assayed, in most cases the highest OTA levels were achieved after 7 days of incubation, whereas this situation occurred at 15 degrees C after 14 days of incubation for only one strain. The maximum OTA level was obtained at earlier growth states when incubation temperature increased. In general, OTA concentration increased as water activity (a(W)) increased with no significant production at 0.85-0.91 a(W) under all temperature levels tested. Production occurred over a range of temperatures (15-30 degrees C) with optimum production at 30 degrees C depending on a(W) assayed. The knowledge of Aspergillus section Nigri ecophysiology is critical in the development and prediction of the risk models of raw material and final product contamination by these species under fluctuating and interacting environmental parameters.

Caracterización fenotípica y genotípica de la resistencia enzimática a las cefalosporinas de tercera generación en Enterobacter spp. / Phenotypic and genotypic characterization of resistance to third-generation cephalosporins in Enterobacter spp.

Enterobacter spp. es un patógeno intrahospitalario que presenta múltiples mecanismos de resistencia a los antibióticos b-lactámicos. Se caracterizaron fenotípica y genotípicamente las diferentes b-lactamasas presentes en 27 aislamientos consecutivos e ininterrumpidos de Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes). También se evaluó la habilidad de diferentes métodos fenotípicos para detectar b-lactamasas de espectro extendido (BLEE) en estos microorganismos. En 15/27 aislamientos (63%) se observó resistencia a las cefalosporinas de tercera generación. En 12 de los aislamientos resistentes se detectó un alto nivel de producción de cefalosporinasa cromosómica, siendo 6 de ellos también productores de PER-2. Dicha resistencia en los 3 aislamientos restantes se debió exclusivamente a la presencia de BLEE, PER-2 en 2 de ellos y CTX-M-2 en un caso. Sólo CTX-M-2 se detectó con todas las cefalosporinas probadas en los ensayos de sinergia, utilizando el método de difusión, mientras que cefepima mejoró la detección de PER-2 en 7/8 aislamientos productores de esta BLEE, 4/8 utilizando la prueba de doble disco y 7/8 comparando discos de cefepima con y sin el agregado de ácido clavulánico. El método de dilución empleado solo detectó 1/9 BLEE al comparar las cefalosporinas con y sin el agregado de inhibidor.

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to b-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum b-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63%) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC b-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor aproximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

[Phenotypic and genotypic characterization of resistance to third-generation cephalosporins in Enterobacter spp]. / Caracterización fenotípica y genotípica de la resistencia enzimática a las cefalosporinas de tercera generación en Enterobacter spp.

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to beta-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63%) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC beta-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor approximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

Caracterización fenotípica y genotípica de la resistencia enzimática a las cefalosporinas de tercera generación en Enterobacter spp. / [Phenotypic and genotypic characterization of resistance to third-generation cephalosporins in Enterobacter spp]

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to beta-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63

) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC beta-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor approximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

[Activity of 14 antimicrobials against Eikenella corrodens]. / Actividad de catorce antimicrobianos sobre Eikenella corrodens.

Eikenella corrodens is a gram-negative bacillus that colonizes as normal flora of the mouth, the upper respiratory tract and the gastrointestinal tract. The aim of this study was to determine the susceptibility patterns against fourteen antibiotics of 25 E. corrodens strains isolated at our hospital. MICs were determined by the agar dilution technique using Müeller-Hinton agar with sheep blood (5% v/v) to penicillin, ampicillin, ampicillin-sulbactam, cephalotin, cefoxitin, ceftiaxone, colistin, gentamicin, amikacin, erythromycin, rifampin, ciprofloxacin and clindamycin. The most active antibiotics were ciprofloxacin and ceftriaxone (MIC90 = 0.008 and 0.125 microgram/ml, respectively), whereas eritromycin, gentamicin and amikacin showed less activity. Only one strain was beta lactamase positive, and it was inhibited by sulbactam. Erithromycin, gentamicin and amikacin had poor activity (MIC90 = 16.8 and 64 micrograms/ml, respectively), whereas all the strains were uniformly resistant to clindamycin (MIC > or = 32 micrograms/ml). We suggest about the need of periodical surveys of E. corrodens susceptibility patterns, since strains have been found with decreased susceptibility against antibiotics which are currently being used for the treatment of infectious diseases.

Actividad de catorce antimicrobianos sobre Eikenella corrodens / [Activity of 14 antimicrobials against Eikenella corrodens]

Eikenella corrodens is a gram-negative bacillus that colonizes as normal flora of the mouth, the upper respiratory tract and the gastrointestinal tract. The aim of this study was to determine the susceptibility patterns against fourteen antibiotics of 25 E. corrodens strains isolated at our hospital. MICs were determined by the agar dilution technique using M³eller-Hinton agar with sheep blood (5

v/v) to penicillin, ampicillin, ampicillin-sulbactam, cephalotin, cefoxitin, ceftiaxone, colistin, gentamicin, amikacin, erythromycin, rifampin, ciprofloxacin and clindamycin. The most active antibiotics were ciprofloxacin and ceftriaxone (MIC90 = 0.008 and 0.125 microgram/ml, respectively), whereas eritromycin, gentamicin and amikacin showed less activity. Only one strain was beta lactamase positive, and it was inhibited by sulbactam. Erithromycin, gentamicin and amikacin had poor activity (MIC90 = 16.8 and 64 micrograms/ml, respectively), whereas all the strains were uniformly resistant to clindamycin (MIC > or = 32 micrograms/ml). We suggest about the need of periodical surveys of E. corrodens susceptibility patterns, since strains have been found with decreased susceptibility against antibiotics which are currently being used for the treatment of infectious diseases.

Actividad de catorce antimicrobianos sobre Eikenella corrodens. / [Activity of 14 antimicrobials against Eikenella corrodens]

Eikenella corrodens is a gram-negative bacillus that colonizes as normal flora of the mouth, the upper respiratory tract and the gastrointestinal tract. The aim of this study was to determine the susceptibility patterns against fourteen antibiotics of 25 E. corrodens strains isolated at our hospital. MICs were determined by the agar dilution technique using M³eller-Hinton agar with sheep blood (5

v/v) to penicillin, ampicillin, ampicillin-sulbactam, cephalotin, cefoxitin, ceftiaxone, colistin, gentamicin, amikacin, erythromycin, rifampin, ciprofloxacin and clindamycin. The most active antibiotics were ciprofloxacin and ceftriaxone (MIC90 = 0.008 and 0.125 microgram/ml, respectively), whereas eritromycin, gentamicin and amikacin showed less activity. Only one strain was beta lactamase positive, and it was inhibited by sulbactam. Erithromycin, gentamicin and amikacin had poor activity (MIC90 = 16.8 and 64 micrograms/ml, respectively), whereas all the strains were uniformly resistant to clindamycin (MIC > or = 32 micrograms/ml). We suggest about the need of periodical surveys of E. corrodens susceptibility patterns, since strains have been found with decreased susceptibility against antibiotics which are currently being used for the treatment of infectious diseases.

[Diagnosis and treatment of Helicobacter pylori infection. Its relationship with gastrointestinal ulcer and antimicrobial resistance]. / Diagnóstico y tratamiento de la infección por Helicobacter pylori. Su relación con la úlcera gastrointestinal y la resistencia a los antimicrobianos.

Reliable data regarding the efficacy of different schemes of triple therapy for the eradication of Helicobacter pylori in our country, are not available. Patients with Helicobacter pylori infection and non-ulcer dyspepsia or active peptic ulcer disease were randomized in three different groups for therapy with, omeprazole 20 mg, clarithromycin 500 mg and amoxicillin 1000 mg, twice daily for one week (OCA 1, 40 patients) and the same treatment but for two weeks in a second group (OCA 2, 40 patients). The third group received omeprazole 20 mg, clarithromycin 500 mg and metronidazole 500 mg twice daily during one week (OCM, 40 patients). The primary efficacy end point was the eradication of Helicobacter pylori as confirmed by negative urea breath test, 4 weeks after the completion of treatment. Of 120 patients enrolled in the study, 113 met the entry criteria. Of them, 103 completed the treatment. When analyzed by intention to treat, after 4 weeks of finishing the treatment, Helicobacter pylori was eradicated in 92.3% of patients in OCA 1, 89.7% in OCA 2, and 82.8% in OCM. There was no significant difference between the three groups, regarding the eradication efficacy. Side effects were observed more frequently in OCA 2 and OCM groups. Primary resistance to amoxicillin and clarithromycin was not demonstrated, while 20% of cultured strains were resistant to metronidazole. In patients with peptic ulcer disease or non-ulcer dysplasia, triple therapy with omeprazole and two antibiotics is highly effective in the eradication of Helicobacter pylori. One week of OCA therapy is as effective as two weeks of OCA or one week of OCM, with less side effects.

Diagnóstico y tratamiento de la infección por Helicobacter pylori. Su relación con la úlcera gastrointestinal y la resistencia a los antimicrobianos. / [Diagnosis and treatment of Helicobacter pylori infection. Its relationship with gastrointestinal ulcer and antimicrobial resistance]

Reliable data regarding the efficacy of different schemes of triple therapy for the eradication of Helicobacter pylori in our country, are not available. Patients with Helicobacter pylori infection and non-ulcer dyspepsia or active peptic ulcer disease were randomized in three different groups for therapy with, omeprazole 20 mg, clarithromycin 500 mg and amoxicillin 1000 mg, twice daily for one week (OCA 1, 40 patients) and the same treatment but for two weeks in a second group (OCA 2, 40 patients). The third group received omeprazole 20 mg, clarithromycin 500 mg and metronidazole 500 mg twice daily during one week (OCM, 40 patients). The primary efficacy end point was the eradication of Helicobacter pylori as confirmed by negative urea breath test, 4 weeks after the completion of treatment. Of 120 patients enrolled in the study, 113 met the entry criteria. Of them, 103 completed the treatment. When analyzed by intention to treat, after 4 weeks of finishing the treatment, Helicobacter pylori was eradicated in 92.3

of patients in OCA 1, 89.7

in OCA 2, and 82.8

in OCM. There was no significant difference between the three groups, regarding the eradication efficacy. Side effects were observed more frequently in OCA 2 and OCM groups. Primary resistance to amoxicillin and clarithromycin was not demonstrated, while 20

of cultured strains were resistant to metronidazole. In patients with peptic ulcer disease or non-ulcer dysplasia, triple therapy with omeprazole and two antibiotics is highly effective in the eradication of Helicobacter pylori. One week of OCA therapy is as effective as two weeks of OCA or one week of OCM, with less side effects.

Cefotaxime-hydrolysing beta lactamases in Morganella morganii.

The frequency of enterobacterial isolates with high resistance to expanded-spectrum beta-lactam antibiotics (mainly cefotaxime or ceftriaxone) has increased notoriously in Argentina, mainly because of the spread of extended-spectrum beta-lactamases. The aim of this work was the study of extended-spectrum beta-lactamases in several Morganella morganii isolates with unusually high resistance to ceftriaxone. These strains produced at least two beta-lactamases, of apparent pIs of 5.4 and 8.2, molecular weight 23 000, well inhibited by clavulanate, compatible with a broad-spectrum beta-lactamase - perhaps TEM-1 - and an extended-spectrum beta-lactamase, respectively. The extended-spectrum beta-lactamase was identified as a CTX-M-type beta-lactamase - probably CTX-M-2 - by polymerase chain reaction, restriction profile analysis and DNA-DNA hybridisation. The remaining isolates studied produced either the broad-spectrum beta-lactamase plus the ubiquitous AmpC beta-lactamase (13 strains), or the AmpC beta-lactamase only (10 strains).