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Enteroaggregative Escherichia coli (EAEC), a biofilm forming pathogen, causes acute and persistent diarrhea worldwide, requiring antimicrobial therapy in severe or persistent cases. To determine the susceptibility of EAEC biofilm to antimicrobials, as single-agent or combined therapy, biofilm formation was investigated using EAEC clinical strains via peg lid. Of the 78 initially analyzed strains, 35 could form biofilms, 15 (42.9%; 15/35) were resistant to at least 1 tested antimicrobial and 20 (57.1%) were susceptible to all of them in the planktonic form. The biofilms of these susceptible strains were challenged against chosen antimicrobials, and displayed resistance to tetracycline, trimethoprim-sulfamethoxazole, chloramphenicol, ampicillin, cefotaxime, ceftriaxone (85%-100%), tobramycin (25%), cefoxitin (20%), and ciprofloxacin (5%). Moreover, ciprofloxacin combined with ampicillin, and tobramycin eradicated the biofilm of 2 of the 4 tested strains. Ciprofloxacin, cefoxitin, and tobramycin maintained their activity well against EAEC biofilm, suggesting their possible effectiveness to treat diarrhea caused by biofilm-forming EAEC strains., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Methicillin-resistant Staphylococcus aureus (MRSA) detection in cystic fibrosis (CF) is challenging. We compared different phenotypic methods among 157 S. aureus from 136 CF-patients: cefoxitin (FOX) and oxacillin (OXA) broth-microdilution; MicroScan-WalkAway®; FOX and OXA disk-diffusion (DD), and PBP2a-latex agglutination. PCR detection of mecA/mecC was the gold standard. Growth on ChromID TM -MRSA agar was evaluated and compared with that of 157 blood culture (BC) isolates. ChromID TM -MRSA was also tested on sputa from 111 CF-patients. 32 isolates (20%) were mecA-positive. Both FOX DD and MicroScan-WalkAway® (FOX/OXA) showed the highest sensitivity and specificity (100% and 100%, 96.9% and 99.2%, 96.9% and 100%). ChromID TM -MRSA showed an excellent sensitivity for BC and CF-isolates (100% and 96.9%) but a poorer specificity for CF ones (95.5% vs. 73.7%), which was also observed when samples were seeded on this medium. FOX DD and MicroScan-WalkAway® are suitable for MRSA detection among CF-isolates and should be used to confirm ChromID TM -MRSA positive CF-cultures., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Linezolid (LZD) serves as an effective option in the treatment of Mycobacterium abscessus complex (MABC) infection. Unfortunately, the combined activities of LZD with other antimicrobial agents against MABC have not been evaluated systemically. In this study, we randomly selected 32 Mycobacterium abscessus and 32 Mycobacterium massiliense isolates for the determination of in vitro synergistic effect between LZD and other antimicrobial agents, including amikacin (AMK), moxifloxacin (MOX), cefoxitin (CFX) and tigecycline (TGC). Out of 64 MABC isolates tested, only one (1.6%, 1/64) and two (3.2%, 2/64) exhibited resistance to AMK and LZD, respectively. Statistical analysis revealed that the percentage of TGC-resistant isolates was significantly lower among M. massiliense (9.4%, 3/32) than that among M. abscessus (25.0%, 8/32, P<0.001). In addition, LZD and AMK showed synergy for 29 MABC isolates (45.3%), whereas no antagonism was noted for this combination. The second mostly frequent synergistic effect was found in LZD plus TGC combination, and 26.6% (17/64) of the strains tested exhibited synergy. In contrast, LZD-CFX and LZD-MOX combinations appeared antagonistic for half of the isolates (48.4%, 31/64 for CFX and 51.6%, 33/64), and almost no synergistic effect was reported in any of the strains for these two combinations. In conclusion, our data reveal that LZD and AMK show the most potent activity against MABC. The frequent synergism is observed in LZD-AMK and LZD-TGC combinations, while LZD rarely exhibits in vitro synergy with MOX and CFX when tested against MABC., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Staphylococcus aureus isolates with discordant susceptibility results between oxacillin and cefoxitin obtained using automated microbiology systems are infrequently observed. From April 2013 to December 2014, 1956 methicillin-resistant S. aureus (MRSA) and 1761 methicillin-susceptible S. aureus isolates were obtained from different patients. Forty isolates (1.1% and 2% in case of S. aureus and MRSA, respectively) with discordant susceptibility results (oxacillin susceptible and cefoxitin resistant) and carrying mecA gene were obtained. Except 2 SCCmec type IV isolates, 38 MRSA isolates were all SCCmec type V (V T or non-V T ), which were further divided into V T (n=22) and non-V T (n=16). The most common spa type in V T and non-V T isolates were t437 (n=19) and t1081 (n=13), respectively. Only 55% of patients received effective antimicrobial agents; 2 mortalities were not attributable to MRSA infection. Using standard agar dilution, 17 MRSA isolates (0.46% and 0.87% in case of S. aureus and MRSA, respectively) had oxacillin MIC in the susceptible ranges (oxacillin-susceptible MRSA [OS-MRSA]); all carried SCCmec type V (V T , n=8; non-V T , n=9). The most common spa-MLST types of OS-MRSA in V T and non-V T were t437-ST59 (n=4) and t1081-ST45 (n=7), respectively. Concomitant testing by both cefoxitin- and oxacillin-based methods is a practical strategy for OS-MRSA detection in the clinical laboratories. Continuous monitoring of OS-MRSA isolates is necessary to elucidate their impact in clinical infectious diseases., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Small-colony variants (SCVs) are slow-growing subpopulations of various auxotrophic bacterial strains. Thymidine-dependent SCVs (TD-SCVs) are unable to synthesize thymidine; hence, these variants fail to grow in a medium without thymidine. In this study, we used 10 TD-SCVs of Staphylococcus aureus, of which four strains possessed mecA. We compared the efficacy of a newly modified medium containing thymidine for the detection of TD-SCVs of methicillin-resistant S. aureus (MRSA) to the efficacy of routinely used laboratory media. We observed that none of the 10 TD-SCVs of S. aureus grew in Mueller-Hinton agar, and four TD-SCVs of MRSA failed to grow on all MRSA screening media, except for the ChromID™ MRSA medium. Laboratory tests conducted using medium with thymidine incorporated showed that thymidine did not affect the minimum inhibitory concentrations of oxacillin and cefoxitin for clinical isolates of S. aureus, and was able to detect MRSA, including TD-SCVs. These findings showed that thymidine-incorporated media are able to detect TD-SCVs of MRSA without altering the properties of other clinically isolated MRSA strains., (Copyright © 2015 Elsevier B.V. All rights reserved.)

During a 1-year period, 87 methicillin-resistant Staphylococcus aureus isolates were collected from 4 major Cuban hospitals for epidemiological analysis. The majority (86%) were related to the community-associated USA300 clone, whereas the remaining belonged to a new clone ST72-V. Interestingly, no hospital-associated clone was found in these Cuban hospitals., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Susceptibility testing on 60 isolates of Staphylococcus epidermidis and 20 isolates of Staphylococcus hominis was performed using the Sensititre (Trek Diagnostics, Cleveland, OH, USA) method and compared with Vitek-2 automated susceptibility testing (bioMèrieux, Marcy l'Etoile, France), MICE strips for minimum inhibitory concentration determination (Oxoid, Basingstoke, UK), and disc diffusion (MAST, Bootle, UK). The categorical agreement between Vitek and Sensititre was greater than 90% for agents tested against S. epidermidis except for tetracycline (52%) and oxacillin (67%), and for S. hominis, the agreement was greater than 90% except for tetracycline (80%), gentamicin (65%), and oxacillin (75%). The categorical agreement between disc diffusion and Sensititre was greater than 90% for antimicrobials tested against S. epidermidis except for tetracycline (45%), moxifloxacin (70%), clindamycin (88%), and cefoxitin (80%), while against S. hominis, the categorical agreement was greater than 90% for antimicrobials tested except for gentamicin (65%), tetracycline (75%), moxifloxacin (80%), and ciprofloxacin (75%). Categorical agreement between MICE strips and Sensititre for S. epidermidis and S. hominis was greater than 95% for vancomycin and daptomycin. Sensititre method shows good categorical agreement with other standard methods for susceptibility testing of S. hominis and S. epidermidis., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Methicillin-resistant Staphylococcus aureus (MRSA) arises when methicillin-susceptible S. aureus (MSSA) acquires the staphylococcal cassette chromosome mec (SCCmec). Most pvl-positive MRSA in Taiwan belong to ST59 lineage and carry SCCmec V. The genetic profiles of 51 MSSA were compared with those of 80 MRSA from the same hospitals. Nine pvl-positive MSSA (oxacillin MIC < or = 2 microg/mL) shared >80% similarity in pulsed field gel electrophoresis pattern with 17 pvl-positive SCCmec V MRSA. Further investigation found that 5 of these 9 isolates were MRSA by cefoxitin and carried SCCmec V. All 26 pvl-positive isolates had very similar genetic profile (ST59, protein A clonal complex [spa-CC] c2:441/437, and agr group I). The success of the ST59:SCCmec V MRSA may be due in part to its heterogeneous and borderline resistance to methicillin, which may be missed by testing only oxacillin, with subsequent exposure to beta-lactams causing the emergence of more resistant subpopulations.

In this study the susceptibility of 58 coagulase-negative staphylococci (CoNS) strains and 58 Staphylococcus aureus strains to oxacillin was evaluated by a novel method called quantitative disk diffusion (DD) method. The results obtained were compared to phenotypic methods as agar dilution (AD) for oxacillin, disk diffusion (DD) for cefoxitin, and related to the presence of the mecA gene detected by PCR. Minimum inhibitory concentrations (MIC) determined by the quantitative DD method were equivalent to MICs determined in the AD method for S. aureus (Student's t test, p=0.99) and CoNS (Student's t test, p=0.97). Incongruent results between PCR mecA gene determinations and the quantitative DD method were obtained in 8 strains (5 S. aureus and 3 CoNS) where the mecA gene expression was blocked. However, oxacillin resistance was detected by the proposed method even in staphylococci strains showing low-level or heterogeneous resistance to the antibiotic while other phenotypic methods failed. The single quantitative DD method is not expensive, it can be performed in any laboratory and permits accurate identification of oxacillin resistant staphylococci.

Phenotypic methods for detecting mecA-mediated resistance in Staphylococcus aureus include both oxacillin and cefoxitin susceptibility tests; many laboratories perform multiple tests. Conflicting oxacillin and cefoxitin susceptibility results are most likely to occur for isolates that either have reduced susceptibility to oxacillin by a non-mecA-mediated mechanism or are mecA positive but are very heteroresistant. To understand the performance of oxacillin and cefoxitin tests for such isolates, we tested 135 S. aureus isolates using either cefoxitin or oxacillin and compared the results with mecA polymerase chain reaction. These strains either expressed borderline oxacillin MICs (1-4 microg/mL) and had undetermined mecA status or were mecA positive but were not detected by oxacillin broth microdilution (BMD) or disk diffusion (DD) in original testing. For 24-h readings, performance of cefoxitin tests (sensitivity/specificity) were DD (99/100), Etest using < or =6 microg/mL as susceptible (99/98), and Phoenix MIC using < or =4 microg/mL as susceptible (98/100). Using 6 microg/mL of cefoxitin as a screen test in both BMD and agar dilution also worked well (98/98-100). Sensitivity/specificity of oxacillin methods were oxacillin agar screen (BBL: 80/86; Remel, Lenexa, KS: 85/50), DD (91/59), BMD (85/88), MicroScan (89/96), VITEK Legacy (82/93), VITEK 2 (91/73), and Phoenix, (67/96). These results suggest that a cefoxitin test can be used alone to predict mecA-mediated resistance in S. aureus.

Detection of plasmid-mediated (P-M) AmpC beta-lactamase-producing isolates is considered critical for epidemiologic studies and hospital infection control, but the documents of the Clinical and Laboratory Standards Institute do not contain any recommendation for the phenotypic detection. In this study, phenotypic detection methods, cefoxitin-Hodge test and induction test, were evaluated using cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae isolates. The cefoxitin-Hodge test detected all bla(CMY-10), and 97.4% of bla(CMY-2) allele-positive isolates, but only 57.3% of bla(DHA-1) allele-positive isolates. Induction test with an aztreonam and an amoxicillin-clavulanic acid disk was more sensitive than with cefoxitin disk, which detected 86.6% of bla(DHA-1) allele-positive isolates. These phenotypic tests should be useful to screen P-M AmpC beta-lactamase-producing E. coli and K. pneumoniae isolates.

Of 176 Salmonella isolates isolated from 2000 to 2003 in a Korean university hospital, 2 cefoxitin-resistant isolates of Salmonella enterica serovar Rissen were found in 2002. Both isolates were resistant to multiple beta-lactam antibiotics, including ampicillin, amoxicillin-clavulanate, cefoxitin, cefotaxime, and ceftazidime. Cell sonicates of the both clinical isolates and their transconjugants had beta-lactamase band of approximate isoelectric point of >8.6. The resistance to cefoxitin was transferred by conjugation to the recipient Escherichia coli J53. The resistance determinant was on conjugative plasmids that encoded a CMY-2, a plasmid-mediated AmpC beta-lactamase. To our knowledge, these are the first description of S. enterica serovar Rissen harboring CMY-2 in South Korea.

One hundred Vitek-2 identified oxacillin resistant S. aureus (ORSA) isolates were tested by oxacillin screen-plate (OXA-plate) and cefoxitin disk (FOX-disk) for confirmation of oxacillin resistance. Ninety-five of 100 confirmed oxacillin resistant by both, 3 of 5 were oxacillin susceptible by both and mecA negative, and 2 of 5 were oxacillin susceptible by OXA-plate, resistant by FOX-disk and mecA positive. The FOX-disk out-performed the OXA-plate for ORSA confirmation and can be performed using standard laboratory techniques.

Cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae are widespread in Korea. Significant proportions of them are considered to be CMY-1 producers. For effective screening of CMY-1 producers, the Hodge test was modified by using a cefoxitin disk and the performance was evaluated. The sensitivity and specifity of the test were 100% and 94.9%, respectively. The test was easier to perform than the three-dimensional extract test. This modified test should be suitable for screening CMY-1-producing strains of E. coli and K. pneumoniae., (Copyright 2002 Elsevier Science B.V.)

Escherichia coli may become resistant to cephamycines and oxyimino cephalosporins by virtue of promotor and attenuator mutations or because they have acquired mobilized beta-lactamases from other gram-negative bacilli. This study examined Canadian strains to determine how often promotor and/or attenuator mutations account for this mechanism of resistance and the extent to which clonal spread of these organisms has occurred. We sequenced the promotor and attenuator region of 30 strains resistant to cefoxitin. Twenty-two strains had promotor mutations, 26 had attenuator mutations. Most promotor mutations resulted either in a change in the -35 promotor region towards the E. coli sigma 70 consensus sequence or in the creation of a new consensus hexamer upstream. Eight strains had mutations that increased the typical ampC 16-nucleotide spacer region to the consensus 17- or an 18-nucleotide sequence. Of the attenuator mutations, most did not substantially affect the attenuator loop. Several of the mutations have previously been described in South Africa, Scandinavia, and France. There was evidence that strains bearing certain mutations were clonally disseminated; however, the 11 strains bearing a complex set of attenuator mutations were not. The majority of cephamycin resistant E. coli strains in Toronto have attenuator and/or promotor mutations upstream of the chromosomal ampC gene.

The objective of this study was to assess the prophylactic efficacy of cefoxitin, ceftizoxime, and metronidazole-gentamicin in colorectal surgery. A double-blind, randomized prospective clinical trial design was used in a Canadian tertiary care teaching hospital. Patients were randomized to one of three treatment groups and received three doses of a study drug (30 min preoperative and 2 postoperative doses at 12 and 24 h). Cefoxitin and ceftizoxime were given as 1000-mg doses. Metronidazole-gentamicin was given as 500 mg of metronidazole plus 120 mg of gentamicin in a minibag. High-risk patients (bowel ischemia, diabetic, current steroid use, etc.) received 10 postoperative doses. Patients with infections, prior antibiotics, or study drug allergies were excluded. Over 30 months, 153 patients were enrolled. Thirty-one patients were excluded for protocol violations. Of the 122 evaluable patients (38 ceftizoxime, 45 metronidazole-gentamicin, 39 cefoxitin), there was no difference across groups regarding sex, age, weight, preoperative Apache II score, and prior history of bowel surgery. Groups were equivalent regarding surgeon, nursing unit, high-risk status (six ceftizoxime, seven metronidazole-gentamicin, seven cefoxitin), bowel preparation, and procedure (including blood loss, drains, organ injury, intraoperative complications). Clinically significant infection requiring systemic antibiotics (7-day hospital and 30-day follow-up) was identified in 0% of ceftizoxime, 15% of metronidazole-gentamicin, and 26% of cefoxitin receiving patients (p = 0.005). Mean ASEPSIS scores for each group were 2.3 (range 0-15) for ceftizoxime, 9.2 (range 0-45) for metronidazole-gentamicin, and 10.4 (range 0-75) for cefoxitin (p = 0.01). Ceftizoxime patients tended to have a shorter total hospital stay (12.2 days versus 19.7 days for cefoxitin versus 13.9 days for metronidazole-gentamicin; p = 0.04), although the procedure to discharge interval was not significantly different (p = 0.09). There was no difference in clinical outcome according to risk status. Anaerobic bacteria were observed more commonly in the ceftizoxime and cefoxitin groups, whereas enteric Gram-negative aerobes were observed most often in the metronidazole-gentamicin group. The study regimens were generally well tolerated. Drug costs were equivalent between ceftizoxime and cefoxitin and lowest with the metronidazole-gentamicin regimen. Ceftizoxime appears to be more effective for the prevention of infection in colorectal surgery than either cefoxitin or metronidazole-gentamicin in the dosage regimens studied.

The inhibitory activity of five beta-lactam agents, alone and in combination with a beta-lactamase inhibitor, was compared with that of cefoxitin and metronidazole against 300 beta-lactamase producing Bacteroides fragilis group isolates. Each of the beta-lactamase inhibitors significantly potentiated the activity of the respective beta-lactam. In the presence of clavulanate, the MIC90 (minimum inhibitory concentration) values of amoxicillin and ticarcillin were reduced 64-fold and 32-fold, respectively. Similarly, sulbactam enhanced the activity of ampicillin and cefoperazone 16-fold and 8-fold, respectively, whereas tazobactam potentiated the activity of piperacillin 16-fold. Few strains were resistant to the beta-lactam-beta-lactamase inhibitor combinations and were comprised of strains of B. fragilis, B. thetaiotamicron, and B. distasonis. Of the strains, 7% were resistant to cefoxitin, and none to metronidazole. Using time-kill kinetic studies, the bactericidal activity of the various beta-lactam agents, with and without beta-lactamase inhibitors, was determined and compared with that of cefoxitin and metronidazole against cefoxitin-susceptible and cefoxitin-resistant isolates of the B. fragilis group. Overall, metronidazole was the most bactericidal agent with all isolates being killed with less than or equal to 4 micrograms/ml at 24 hr. Ampicillin-sulbactam was the next most bactericidal agent with all isolates being killed with less than or equal to 16/8 micrograms/ml of ampicillin-sulbactam at 24 hr. Amoxicillin-clavulanate and cefoperazone-sulbactam had bactericidal activity similar to that of ampicillin-sulbactam. Piperacillin-tazobactam and ticarcillin-clavulanate were bactericidal at higher concentrations with all isolates killed with 64 micrograms/ml of piperacillin and 128 micrograms/ml of ticarcillin combined with their respective beta-lactamase inhibitors. None of the beta-lactam agents alone was able to kill more than 19 of the 26 isolates. We conclude that beta-lactam agents combined with beta-lactamase inhibitors have both inhibitory and bactericidal activity against cefoxitin-resistant members of the B. fragilis group provided that the concentrations achieved for these combinations are at the upper limits for maximum recommended dosing. Although isolates of the B. fragilis group have been reported to produce unusual beta-lactamases that are refractory to beta-lactamase inhibitors, none of the cefoxitin-resistant isolates tested in this study were resistant to the beta-lactam-beta-lactamase inhibitor combinations.

Time-kill kinetic methodology was used to evaluate the bactericidal activity of cefoxitin, cefotetan, clindamycin, and metronidizole against cefoxitin-susceptible and cefoxitin-resistant isolates of the Bacteroides fragilis group. Overall, metronidazole was the most bactericidal agent, with all isolates being killed with less than or equal to 4 micrograms/ml at 24 hr. Clindamycin was the next most bactericidal agent, with 20 of 26 isolates being killed with less than 16 micrograms/ml. Six isolates with clindamycin MICs greater than or equal to 64 micrograms/ml were not killed at 24 hr, with concentrations as high as 256 micrograms/ml. Cefoxitin and cefotetan were the least bactericidal agents tested. Seven isolates with MICs of greater than or equal to 64 micrograms/ml to each agent demonstrated a lack of killing at 24 hr, with concentrations of the respective agent as high as 256 micrograms/ml. At concentrations with either agent of 32 micrograms/ml, the remaining 19 isolates were killed at 24 hr. Of the six B. fragilis isolates resistant to clindamycin, four were also resistant to both cefoxitin and cefotetan. We conclude that in hospitals with cefoxitin-resistant B. fragilis group isolates, metronidazole would provide appropriate therapy.

Ceftibuten, like cefotaxime, was observed to be a weak inducer of the class-I beta-lactamases of Enterobacter cloacae, Serratia marcescens, and Enterobacter aerogenes. In contrast, cefoxitin and imipenem induced these enzymes strongly at subinhibitory concentrations.

Cefotaxime, cefoxitin, ceftazidime, and cefuroxime were tested in a multicenter study to establish susceptibility testing criteria and quality control guidelines for Neisseria gonorrhoeae. Interpretive criteria were established by using triplicate testing of at least 100 gonococcal strains having various susceptibility patterns to currently utilized drug regimens. Only a susceptible category was proposed for cefotaxime and ceftazidime (greater than or equal to 31 mm and less than or equal to 0.5 micrograms/ml) because of rare resistant isolates. The other interpretive crtieria were cefoxitin susceptible, greater than or equal to 28 mm (less than or equal to 2 micrograms/ml); intermediate, 24-27 mm (4 micrograms/ml), and resistant, less than or equal to 23 mm (greater than or equal to 8 micrograms/ml); cefuroxime-susceptible, greater than or equal to 31 mm (less than or equal to 1 micrograms/ml); moderately susceptible, 26-30 mm (2 micrograms/ml); and resistant, less than or equal to 25 mm (greater than or equal to 4 micrograms/ml). Quality control criteria were established by using multiple agar lots, three disk lots, and a number of replicates consistent with National Committee for Clinical Laboratory Standards M23-T guidelines.

Nineteen strains of the Bacteroides fragilis group were used to determine the bactericidal activity of ceftizoxime, cefoxitin, cefotetan, and penicillin G with time-kill kinetics studies. Each antimicrobial agent was tested at subinhibitory (1/2 X MIC), inhibitory (1 X MIC), and suprainhibitory (4 X MIC) concentrations. Penicillin G exhibited virtually no sustained bactericidal activity at any of the antimicrobial concentrations tested. At subinhibitory concentrations, ceftizoxime was considerably more bactericidal than cefoxitin or cefotetan: At 12 hr, ceftizoxime killed 89% of the inoculum, whereas cefoxitin and cefotetan killed 35% and 33% of the inoculum, respectively. At inhibitory concentrations, ceftizoxime was again more bactericidal than cefoxitin and cefotetan: At 12 hr, ceftizoxime killed 90% of the inoculum, whereas cefoxitin and cefotetan killed 78% and 73%, respectively. At suprainhibitory concentrations, all three antimicrobial agents showed comparable bactericidal activity at 12 and 24 hr. Ceftizoxime and cefoxitin had somewhat lower killing rates overall against test strains with high MICs (greater than or equal to 32) versus low MICs (less than or equal to 16). However, at subinhibitory concentrations, ceftizoxime killed the B. fragilis group strains with high or low MIC values more effectively than cefotetan killed strains with low MICs. At the highest antibiotic concentrations tested (4 X MIC), only slight differences were seen in the bactericidal activity of the three compounds, regardless of MICs.

Three new beta-lactams were evaluated against 94 anaerobic strains representing 15 species using a Wilkins-Chalgren broth microdilution method. The penems, Sch 29482 and Sch 34343, were most active with all minimum inhibitory concentrations (MICs) at less than or equal to 4.0 micrograms/ml and MIC90s of less than or equal to 0.25 micrograms/ml. BMY 28142 had a more limited antianaerobic activity against Bacteroides fragilis with a MIC50 and MIC90 of 32 and 128 micrograms/ml, respectively. Cefbuperazone (T-1982) had low B. fragilis MICs (MIC90, 8.0 micrograms/ml), but potentially resistant range MIC90 results for the other species in the B. fragilis group and Clostridium species.

Ceftizoxime and cefoxitin (control) were tested against 99 anaerobic bacteria using three commonly used susceptibility testing methods. The cefoxitin MIC results with the National Committee for Clinical Laboratory Standards (NCCLS) reference agar dilution (RAD) method and the broth microdilution (BMD) method did not differ by more than one log2 dilution step. However, the ceftizoxime BMD MIC results were 2- to 4-fold lower than those produced by the RAD procedure. Broth disk elution (BDE) tests for both cephalosporins (ceftizoxime and cefoxitin) had very high rates of false-susceptible and false-resistant error when compared to the RAD MICs: the BDE tests are not recommended for either cephalosporin. Findings indicate that because of method and possible medium or technical variability, ceftizoxime MICs should be determined only by the NCCLS RAD method. Also, cefoxitin susceptibility should be assessed primarily by RAD or BMD procedures. This should minimize confusion related to ceftizoxime's spectrum and activity against anaerobic bacteria compared to similar beta-lactam drugs. Ceftizoxime quality control guidelines were established for Bacteroides fragilis (ATCC 25285): mode, 64 micrograms/ml, range 32-128 micrograms/ml. Other control strains were not reliable.

Three aminothiazolyl methoxyimino cephalosporins (ceftizoxime, ceftriaxone, and cefotaxime with and without its metabolite) were tested against 500 strains of anaerobic bacteria using the NCCLS reference agar dilution procedure. Eighty-seven percent of all strains tested were from the Bacteroides fragilis group. When tested against a collection of 100 cefoxitin-resistant isolates, ceftizoxime and the cefotaxime/desacetyl-cefotaxime combination were the most active in vitro, inhibiting 32-38% of strains. Ceftriaxone inhibited the greatest number (87%) of cefoxitin-susceptible anaerobes at less than or equal to 32 micrograms/ml. A regional variation in the activity of these drugs was confirmed when an additional 300 isolates were examined from three medical centers. Cefotaxime, ceftizoxime, and ceftriaxone were essentially equal in overall antimicrobial activity, although each drug was judged the best at one of the three locations. Cefoxitin resistance (MIC greater than or equal to 32 micrograms/ml) was consistent among the institutions at a 25% incidence for all organisms tested, and 28% cefoxitin resistance among the B. fragilis group strains. Cefoxitin resistance was not determined to be associated with a beta-lactamase mechanism, but ceftriaxone and other aminothiazolyl cephems were hydrolyzed, thus elevating their MICs. Regional variation in anaerobic organism susceptibility to cephamycins and cephalosporins in company with the variable beta-lactam resistance mechanisms seems to require periodic, epidemiologic monitoring of in vitro drug activity by appropriate methods to assure continued antimicrobial efficacy.

Cefotaxime (CTX) and desacetylcefotaxime (des-CTX) alone and in combination were tested against anaerobic bacteria collected from clinical infections from several geographically diverse medical centers. The CTX minimum inhibitory concentration (MIC) inhibiting 50% of tested Bacteroides fragilis strains was in the moderately susceptible range (32 micrograms/ml), but when placed in combination with des-CTX it had a potency compatible to cefoxitin (MIC50, 8.0 micrograms/ml). Other B. fragilis group species (B. distasonis and B. vulgatus) were also susceptible to CTX and des-CTX alone at the MIC50 level. MIC90 statistics for CTX, cefoxitin, and ticarcillin were generally in the resistant range. Synergy studies showed that 80% of tested anaerobes were synergistically killed by the combination of CTX and des-CTX. Most of these strains had their synergy occur at drug levels that could be achieved in vivo. A large number of the B. thetaiotaomicron strains must be considered resistant to the combination because of the very high levels of des-CTX required to produce synergistic killing. Other drugs routinely used for anaerobic infections (clindamycin, chloramphenicol, and metronidazole) also had elevated B. thetaiotaomicron MICs. Endemic difference in susceptibility to the beta-lactam drugs were observed, especially the CTX-des-CTX combination. The combination and other beta-lactams were most usable for strains isolated from the Portland metropolitan area, were moderately active against those from Cleveland, and were rarely usable on Bacteroides isolates at Northwestern in Chicago. Laboratories are urged to monitor the cephalosporin and semisynthetic penicillin in vitro efficacy and not rely on published statistics. Staphylococcus aureus strains were susceptible to CTX alone, but were even more susceptible (two- to fourfold reduction in MICs) when used in combination with des-CTX. These data show CTX to be the most active antistaphylococcal compound among the new cephalosporins and to be comparable to cefamandole and cefuroxime, but superior to the anaerobe-active cefoxitin.

The comparative in vitro activity of cefoxitin, piperacillin, cefotetan, and ceftizoxime was compared against 843 surgical microbial isolates. All compounds exhibited excellent activity against the streptococcal isolates. Antimicrobial activity was poor for Staphylococcus aureus and S. epidermidis. Cefotetan activity against the Enterobacteriaceae was comparable to ceftizoxime (greater than 95% susceptible). Resistance rates of 0, 1, 5, and 2% were observed with cefoxitin, piperacillin, cefotetan, and ceftizoxime against the anaerobic cocci and anaerobic gram-positive non-spore-forming rods. Ninety-six and ninety-nine percent of the clostridial strains were susceptible to cefotetan and piperacillin, respectively. Piperacillin, cefotetan, and ceftizoxime exhibited similar activity against Bacteroides fragilis (resistance less than 10%). Cefotetan and cefoxitin exhibited poor activity against Bacteroides distasonis, B. ovatus, and B. thetaiotaomicron. Antimicrobial activities were comparable for the four drugs against other Bacteroides and Fusobacterium species. The results demonstrate that all four compounds exhibited broad antimicrobial activity against facultative and obligate anaerobic surgical isolates from intraabdominal and soft tissue infections.

The new aminothiazoyl-cephalosporin, ceftriaxone (Ro 13-9904), was found to have excellent inhibitory activity against the Enterobacteriaceae (minimum inhibitory concentration needed to inhibit 50% of isolates (MIC50) less than or equal to 0.004-0.5 microgram/ml, Haemophilus influenzae (MIC50 less than or equal to 0.004 micrograms/ml), Neisseria species (MIC50 less than or equal to 0.001 microgram/ml), pneumococci (MIC50 0.25 micrograms/ml), Staphylococcus aureus (MIC50 2.0 micrograms/ml), and Streptococcus pyogenes (MIC50 0.015 micrograms/ml). Ceftriaxone was less effective against Acinetobacter species, Pseudomonas aeruginosa, and other Pseudomonas species (MIC50 8.0-16 micrograms/ml). Methicillin-resistant S. aureus and enterococci were not significantly inhibited by ceftriaxone. Ceftriaxone was very resistant to beta-lactamase hydrolysis, although the type IV cephalosporinase minimally destroyed the compound at 16.4-19.9% of the rates for cephaloridine. Type I cephalosporinases were inhibited by ceftriaxone and related enzyme-stable cephalosporins. Based on analysis of disk-MIC regression statistics, tentative recommendations for the disk test of the National Committee for Clinical Laboratory Standards are 21 mm or more = susceptible, 14-20 mm = moderately susceptible, and 13 mm or less = resistant. These criteria produce interpretive accuracy of more than 92%, with very rare major errors. Ceftriaxone was comparable to cefotaxime in spectrum and activity, thus allowing the use of the "spectrum-class" concept (for example, cefotaxime tests in vitro to predict ceftriaxone susceptibility, and vice versa).

Amdinocillin alone and in combination with other beta-lactam antibiotics was tested for in vitro activity against aminoglycoside-susceptible and resistant gram-negative bacteria. Amdinocillin alone or in combination with ampicillin, ticarcillin, piperacillin, cefazolin, cefoxitin, and cefamandole had little to no activity against aminoglycoside-resistant E. coli, E. cloacae, K. pneumoniae, and S. marcescens. There was better activity with aminoglycoside-susceptible organisms, however, Overall, there was significantly more antagonism of amdinocillin combinations when tested with aminoglycoside-resistant organisms than with aminoglycoside-susceptible strains.

The agar dilution method was used to determine the inhibitory activity of cefotaxime (CTX) alone, desacetylcefotaxime (dCTX) alone, CTX plus dCTX, and cefoxitin against 74 clinical isolates of the Bacteroides species recovered from diabetic patients with foot ulcers. The study concluded that the addition of dCTX to CTX increased the inhibitory activity from 45% to 73% for all strains tested and from 50% to 81% among the 32 strains of Bacteroides fragilis. This synergistic interaction against B. fragilis resulted in a four- to nine-fold reduction in the MIC of seven strains (64-128 micrograms/ml, resistant category MICs). While the lowest CTX MIC for B. fragilis was 2 micrograms/ml (four strains), the addition of dCTX also produced a remarkable reduction in susceptible range CTX MICs to 0.05-2 micrograms/ml in 16 strains (50%). The overall susceptibility to cefoxitin and CTX plus dCTX was as follows: 100% and 100% for Bacteroides vulgatus, 50% and 66% for Bacteroides thetaiotaomicron, 100% and 33% for Bacteroides ovatus, and 83% and 82% for Bacteroides species other than the B. fragilis group.

The synergistic activity of the combination cefotaxime-desacetylcefotaxime (CTX/dCTX) was compared to the effectiveness of seven other antimicrobial agents: cefoxitin (CFOX), cefotetan (CTAN), ceftizoxime (CTIZ), chloramphenicol (CLOR), clindamycin (CLIND), metronidazole (METR), and ampicillin-sulbactam (A/S) tested against 100 clinical isolates belonging to the Bacteroides fragilis group. All tests were performed using the NCCLS reference agar-dilution method. The overall susceptibility of these organisms to CTX/dCTX was 84% compared to CFOX at 78% or CTAN at 66%. The other antimicrobials inhibited greater than 90% of these isolates. There was no difference between the susceptibility rates of CTX/dCTX and CTX with the B fragilis (85%) or B. distasonis (75%) strains. Bacteroides thetaiotaomicron showed a 11% greater susceptibility to CTX/dCTX than to CTX. Of the 100 isolates tested, 40% showed either synergy or partial synergistic interactions between CTX and dCTX. Most of the isolates showed indifference (52%), while 8% demonstrated antagonism; a relatively unique finding to date.

The antimicrobial activity of three 7-alpha-methoxy beta-lactams were compared to cefoperazone and ceftriaxone. All had a similar spectrum of activity against the Enterobacteriaceae, except cefoxitin. Cefotetan was only slightly less active than moxalactam against the Enterobacter spp. Ceftriaxone was most effective on Neisserias, Haemophilus spp, nonenterococcal Streptococcus spp, and Acinetobacter spp. Cefoperazone generally inhibited more pseudomonads while all of the "cephamycins" showed activity against Bacteroides fragilis and B. thetaiotaomicron. Beta-lactamase hydrolysis studies of six substrates having pharmacologic serum half lives of greater than or equal to 2 hrs were performed by bioassay and automated procedures. Excellent correlations were found between methods up to 24 hrs. A "lag-phase" was observed for several drug/enzyme combinations before initiation of significant substrate hydrolysis. The 7-alpha-methoxy beta-lactams were routinely more stable to the six representative enzymes (Richmond-Sykes types I-V) than other "stable" cephalosporins. Substrate hydrolysis rates resulting in greater than 50% drug loss in less than or equal to 1 hr generally produced resistant in vitro test results. Cefotetan, cefoxitin, moxalactam, ceftriaxone, and dicloxacillin were potent inhibitors of Type I (P99) beta-lactamases. Moxalactam demonstrated significant inhibition and affinity for the Type V enzyme while cefoperazone uniquely possesses affinity (so-called inhibition) for all tested beta-lactamases. Cefotetan appears to be a promising, beta-lactam compound with some in vitro characteristics comparable to the 1-oxa-beta-lactams and alpha-methoxyimino cephalosporins.

Two hundred and forty-six strains of the Bacteroides fragilis group, all clinical isolates, collected at the Wadsworth Veterans Administration Medical Center from 1977 to 1982, were tested for susceptibility to clindamycin and cefoxitin. There was no significant change in resistance to either clindamycin or cefoxitin over the time period tested for any individual species, nor for the B. fragilis group in toto. Striking differences in susceptibility to the two drugs were seen among species of the B. fragilis group. B. fragilis displayed resistance to cefoxitin (32 micrograms/ml) and clindamycin (8 micrograms/ml) of 0.0% and 0.8%, respectively, whereas B. thetaiotaomicron showed resistances of 12.7% to cefoxitin (32 micrograms/ml) and 9% to clindamycin (8 micrograms/ml). B. thetaiotaomicron, B. distasonis, and B. ovatus are distinctly more resistant to cefoxitin than B. fragilis and B. vulgatus. Similarly B. thetaiotaomicron and B. distasonis are much more resistant to clindamycin than are the other B. fragilis group species. It is apparent that determination of species within the B. fragilis group is important in evaluating a potential therapeutic regimen.

Resistance to cefoxitin among species of the Bacteroides fragilis group of organisms has remained low (8%-10%) in a multicenter nationwide survey. However, a statistically significant increase in the percentage of B. fragilis group organisms resistant to cefoxitin was found at Tufts-New England Medical Center from 1981 to 1982. Non fragilis species accounted for most of the resistance. The presence of cefoxitin resistance in B. fragilis isolates correlated with resistance to other antibiotics. The presence of cefoxitin-resistant B. fragilis group organisms also correlated with the presence of other cefoxitin-resistant bacteria. No difference could be detected in therapeutic outcome of patients with cefoxitin-sensitive or cefoxitin-resistant B. fragilis group organisms, regardless of treatment with cefoxitin or other antibiotics.