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Microorganisms Dec 2022Background microorganism growth on Chromogenic Coliform Agar (CCA) can be challenging. For this reason, a new alternative method with a Cefsulodin/Vancomycin...
Background microorganism growth on Chromogenic Coliform Agar (CCA) can be challenging. For this reason, a new alternative method with a Cefsulodin/Vancomycin (CV)-supplemented CCA should be developed in this study. CCA supplemented with CV was validated according to ÖNORM EN ISO 16140-4:2021 using water from natural sources in Styria, Austria. Results show that the alternative method using the supplemented CCA has similar values in relation to sensitivity (82.2%), specificity (98.6%) and higher selectivity (59%) compared to the reference method. Repeatability and reproducibility were acceptable for the alternative method and showed similar results with the reference method. The alternative method shows a very low false positive rate and a low false negative rate paired with good performance regarding the inclusion study. The exclusion study shows the advantage of our method by suppressing background microorganisms and facilitating the process of enumeration of and other coliform bacteria on CCA plates. and growth was inhibited using the supplement. To conclude, the coliform CV selective supplement combined with CCA is an appropriate tool for coliform bacteria detection in water samples.
PubMed: 36557752
DOI: 10.3390/microorganisms10122499 -
Antimicrobial Agents and Chemotherapy Mar 2024Non-clinical antibiotic development relies on susceptibility and infection model studies. Validating the achievement of the targeted drug concentrations is essential to...
Non-clinical antibiotic development relies on susceptibility and infection model studies. Validating the achievement of the targeted drug concentrations is essential to avoid under-estimation of drug effects and over-estimation of resistance emergence. While certain β-lactams (e.g., imipenem) and β-lactamase inhibitors (BLIs; clavulanic acid) are believed to be relatively unstable, limited tangible data on their stability in commonly used media are known. We aimed to determine the thermal stability of 10 β-lactams and 3 BLIs via LC-MS/MS in cation-adjusted Mueller Hinton broth at 25 and 36°C as well as agar at 4 and 37°C, and in water at -20, 4, and 25°C. Supplement dosing algorithms were developed to achieve broth concentrations close to their target over 24 h. During incubation in broth (pH 7.25)/agar, degradation half-lives were 16.9/21.8 h for imipenem, 20.7/31.6 h for biapenem, 29.0 h for clavulanic acid (studied in broth only), 23.1/71.6 h for cefsulodin, 40.6/57.9 h for doripenem, 46.5/64.6 h for meropenem, 50.8/97.7 h for cefepime, 61.5/99.5 h for piperacillin, and >120 h for all other compounds. Broth stability decreased at higher pH. All drugs were ≥90% stable for 72 h in agar at 4°C. Degradation half-lives in water at 25°C were >200 h for all drugs except imipenem (14.7 h, at 1,000 mg/L) and doripenem (59.5 h). One imipenem supplement dose allowed concentrations to stay within ±31% of their target concentration. This study provides comprehensive stability data on β-lactams and BLIs in relevant media using LC-MS/MS. Future studies are warranted applying these data to antimicrobial susceptibility testing and assessing the impact of β-lactamase-related degradation.
Topics: beta-Lactamase Inhibitors; beta-Lactams; Doripenem; Agar; Chromatography, Liquid; Tandem Mass Spectrometry; Anti-Bacterial Agents; Penicillins; Clavulanic Acid; Imipenem; Water; Microbial Sensitivity Tests
PubMed: 38329330
DOI: 10.1128/aac.01399-23 -
The Journal of Antibiotics Jun 1978A single dose of 20 mg/kg of cefsulodin [3-(4-carbamoyl-1-pyridiniomethyl)-7 beta-(D-alpha-sulfophenylacetamido)-ceph-3-em-4-carboxylate monosodium salt] was...
A single dose of 20 mg/kg of cefsulodin [3-(4-carbamoyl-1-pyridiniomethyl)-7 beta-(D-alpha-sulfophenylacetamido)-ceph-3-em-4-carboxylate monosodium salt] was administered subcutaneously to mice, and intramuscularly to rats and dogs. The plasma and tissue levels reached the peak 15 approximately 30 minutes after administration. In mice and rats, no plasma levels were measurable 2 and 4 hours after administration. In dogs, the plasma levels were measurable 6 hours after administration. The level in the kidney of mice was slightly lower than the plasma level, while in rats and dogs, the level in the kidney was higher than the plasma level. The cefsulodin levels in the lung of rats and dogs were relatively high, and the level in mice was relatively low. The hepatic levels were very low in all test animal species. Cefsulodin was mainly excreted into the urine, and the excretion of cefsulodin into the bile was very slight.
Topics: Animals; Bile; Cephalosporins; Dogs; Injections, Intramuscular; Injections, Subcutaneous; Intestinal Absorption; Male; Mice; Mice, Inbred Strains; Rats; Species Specificity; Time Factors
PubMed: 681241
DOI: 10.7164/antibiotics.31.593 -
Frontiers in Immunology 2020Leukocyte inflammatory responses require integrin cell-adhesion molecule signaling through spleen tyrosine kinase (Syk), a non-receptor kinase that binds directly to...
Leukocyte inflammatory responses require integrin cell-adhesion molecule signaling through spleen tyrosine kinase (Syk), a non-receptor kinase that binds directly to integrin β-chain cytoplasmic domains. Here, we developed a high-throughput screen to identify small molecule inhibitors of the Syk-integrin cytoplasmic domain interactions. Screening small molecule compound libraries identified the β-lactam antibiotics cefsulodin and ceftazidime, which inhibited integrin β-subunit cytoplasmic domain binding to the tandem SH2 domains of Syk (IC range, 1.02-4.9 µM). Modeling suggested antagonist binding to Syk outside the pITAM binding site. Ceftazidime inhibited integrin signaling Syk, including inhibition of adhesion-dependent upregulation of interleukin-1β and monocyte chemoattractant protein-1, but did not inhibit ITAM-dependent phosphorylation of Syk mediated by FcγRI signaling. Our results demonstrate a novel means to target Syk independent of its kinase and pITAM binding sites such that integrin signaling this kinase is abrogated but ITAM-dependent signaling remains intact. As integrin signaling through Syk is essential for leukocyte activation, this may represent a novel approach to target inflammation.
Topics: Anti-Inflammatory Agents; Cefsulodin; Ceftazidime; High-Throughput Screening Assays; Humans; Integrin beta Chains; Leukocytes; Male; Phosphorylation; Protein Binding; Protein Interaction Domains and Motifs; Signal Transduction; Small Molecule Libraries; Syk Kinase; THP-1 Cells
PubMed: 33488575
DOI: 10.3389/fimmu.2020.575085 -
Applied and Environmental Microbiology Jun 1996Cefsulodin was evaluated as a potential selective agent for aeromonads. Resistance of Aeromonas and coliform isolates was determined by using a standard disk diffusion... (Comparative Study)
Comparative Study
Cefsulodin was evaluated as a potential selective agent for aeromonads. Resistance of Aeromonas and coliform isolates was determined by using a standard disk diffusion technique. A total of 119 Aeromonas and 78 coliform strains were isolated. For 102 of 130 [corrected] Aeromonas isolates (environmental and reference strains), the MIC of cefsulodin was < 8 micrograms/ml. Results of MIC tests by the agar dilution method showed that a concentration of cefsulodin of 10 micrograms/ml or less inhibited the growth of 96% of isolates. In comparison, for 81 of 94 coliform isolates (environmental and reference strains), the MIC of cefsulodin was > 32 micrograms/ml. Because cefsulodin suppresses growth of Aeromonas and other oxidase-positive organisms, total coliform (TC) and Escherichia coli counts on Chromocult Coliform agar (CC agar) without cefsulodin and on CC agar with 10 mg of cefsulodin per liter (CC-CFS) were compared. Variance analysis of data from 14 sewage-polluted irrigation water specimens did not demonstrate any statistically significant difference in the enumeration of E. coli with CC and CC-CFS media. On average, the CC agar recovered 2.46 times as many TCs as CC-CFS. However, Aeromonas colonies made up an average of 58.6% of the TC counts on CC agar. Because no Aeromonas spp. were recovered on CC-CFS, background interference was eliminated and the counts that were obtained reflected more accurately the number of TCs. Results of this study suggest that cefsulodin may be a useful selective agent against Aeromonas spp. which should be included in coliform chromogenic media when high levels of accompanying flora are expected.
Topics: Aeromonas; Cefsulodin; Cephalosporins; Colony Count, Microbial; Culture Media; Drug Resistance, Microbial; Enterobacteriaceae; Escherichia coli; Microbial Sensitivity Tests; Sewage; Water Microbiology
PubMed: 8787387
DOI: 10.1128/aem.62.6.1885-1888.1996 -
European Journal of Biochemistry Feb 1989A pair of strains of Pseudomonas aeruginosa (3-Pre: cefsulodin-sensitive, inducible beta-lactamase; and 3-Post: cefsulodin-resistant, elevated beta-lactamase, derived...
Permeability to cefsulodin of the outer membrane of Pseudomonas aeruginosa and discrimination between beta-lactamase-mediated trapping and hydrolysis as mechanisms of resistance.
A pair of strains of Pseudomonas aeruginosa (3-Pre: cefsulodin-sensitive, inducible beta-lactamase; and 3-Post: cefsulodin-resistant, elevated beta-lactamase, derived from 3-Pre by subculture in the presence of cefsulodin) were taken as representative of the class of bacteria resistant to third-generation cephalosporins due to elevated synthesis of the normally inducible, chromosomally encoded beta-lactamase. These two strains were used to differentiate between 'trapping' and 'hydrolytic' mechanisms of cefsulodin resistance by (a) measuring the outer-membrane permeabilities to cefsulodin, (b) measuring the kinetics of cefsulodin hydrolysis and the stoichiometry of cefsulodin trapping by the periplasmic beta-lactamase, and (c) comparing the predictions of the trapping and hydrolysis hypotheses with the minimum inhibitory concentrations (MIC) of cefsulodin. The MIC of cefsulodin for strains 3-Pre and 3-Post were 2.35 microM (1.25 micrograms ml-1) and 37.6 microM (20.0 micrograms ml-1) respectively. The permeability parameter for cefsulodin of the outer membrane of the resistant strain was 0.0034 cm3 min-1 mg dry mass-1, so the flux of cefsulodin across its outer membrane at the MIC was calculated to be 0.120 nmol min-1 mg dry mass-1. Hydrolysis of cefsulodin by the beta-lactamase in the periplasm occurred at a rate of 0.118 nmol min-1 mg dry mass-1 which can thus account for resistance by matching the above rate of inflow. Trapping by the beta-lactamase, even with a 1:1 stoichiometry, would require the enzyme to be synthesized at 5.0 micrograms protein min-1 mg dry mass-1 or about 40% of the dry mass/generation. We conclude that hydrolysis, but not trapping, adequately explains the resistance to cefsulodin in P. aeruginosa 3-Post. A similar calculation for latamoxef resistance, using data taken from the literature, led to the same conclusion.
Topics: Bacterial Outer Membrane Proteins; Cefsulodin; Cell Membrane Permeability; Drug Resistance; Enzyme Induction; Hydrolysis; Isoelectric Focusing; Kinetics; Plasmids; Pseudomonas aeruginosa; beta-Lactamases
PubMed: 2493375
DOI: 10.1111/j.1432-1033.1989.tb14599.x -
Journal of Applied Microbiology Jun 2006In this study, the growth characteristics of Yersinia enterocolitica biotype 4, GER O:3 plasmid bearing (P+) and plasmid cured (P-) strain types were evaluated in brain...
AIM
In this study, the growth characteristics of Yersinia enterocolitica biotype 4, GER O:3 plasmid bearing (P+) and plasmid cured (P-) strain types were evaluated in brain heart infusion broth supplemented with cefsulodin, irgasan, and novobiocin alone or in combination.
METHODS AND RESULTS
Growth curves were obtained for the two strain types in broth supplemented with selective agents at 25 or 37 degrees C for 32 h to obtain data on the lag phase durations and growth rates of the strains. Generally, the lag times and growth rates of the P+ and P- strains were similar for cultures incubated at 25 degrees C regardless of the selective agent added and where plasmid replication and expression were not under any significant burden. However, where the lag times and growth rates of the strains were examined at 37 degrees C, significant differences were observed in the lag phase durations of the plasmid bearing strain type compared the plasmid cured strain, an effect that was due to the burden of the plasmid and the influence of selective agents. Generally, when two or more agents were present, lag phase durations were longer for the plasmid bearing strain. Some exceptions noted where in the presence of irgasan or full selective agent (CIN) the opposite case was observed. When growth rates were compared, the plasmidless strain type was typically faster than the plasmid bearing strain in the presence of most selective agents at 37 degrees C and the growth rates of both strain types at 25 degrees C were similar where the temperature appeared to negate the effects of plasmid.
CONCLUSIONS
The data obtained in these studies suggest that selective agents (in particular irgasan) and incubation temperature play a significant role in influencing the growth characteristics of plasmid bearing and plasmid cured strains of Y. enterocolitica.
SIGNIFICANCE AND IMPACT OF THE STUDY
This data presented in this study has significant implications for enrichment methods used in the detection or recovery of plasmid bearing Y. enterocolitica strains from food, environmental or clinical samples.
Topics: Anti-Bacterial Agents; Bacteriology; Carbanilides; Cefsulodin; Culture Media; Food Microbiology; Hot Temperature; Hydrogen-Ion Concentration; Novobiocin; Plasmids; Yersinia enterocolitica
PubMed: 16696677
DOI: 10.1111/j.1365-2672.2006.02861.x -
The Journal of Antibiotics Oct 1981The effect of cefsulodin in combination with various beta-lactam antibiotics was examined against Serratia marcescens. In vitro, the optimum ratio for all combinations...
The effect of cefsulodin in combination with various beta-lactam antibiotics was examined against Serratia marcescens. In vitro, the optimum ratio for all combinations tested was almost the same (cefsulodin - other antibiotic = 1:1 approximately 1:4). The combinations of cefsulodin-cefazolin and cefsulodin-cefotiam were found to have a synergistic effect and other combinations, such as cefsulodin-cefmenoxime, -ampicillin and -sulbenicillin, an additive effect with the checkerboard dilution and the fixed combination methods. The synergistic effect of cefsulodin-cefotiam was more potent than that of cefsulodin-cefazolin and the effect of both combinations was clearer with heavy than with light inoculum size. With the killing kinetic method, all combinations tested showed a synergistic effect. In vivo, the optimum combination ratios of cefsulodin-cefazolin and cefsulodin-cefotiam were 1:2 and 1:1, respectively, the protective effect of the latter combination being much stronger than that of the former. With the fixed combination method (cefsulodin - other antibiotic = 1:1 approximately 1:4), the effect of the combination of cefsulodin with all antibiotics except cefazolin and cefotiam was additive.
Topics: Animals; Anti-Bacterial Agents; Cefsulodin; Cephalosporins; Drug Synergism; Enterobacteriaceae Infections; Male; Mice; Mice, Inbred ICR; Serratia marcescens
PubMed: 7031032
DOI: 10.7164/antibiotics.34.1327 -
Veterinary Microbiology Jun 2022A Yersinia pseudotuberculosis outbreak was diagnosed in a male turkey flock in Finland. Y. pseudotuberculosis is a quite rare zoonotic bacterium, which typically causes...
A Yersinia pseudotuberculosis outbreak was diagnosed in a male turkey flock in Finland. Y. pseudotuberculosis is a quite rare zoonotic bacterium, which typically causes enteritis in humans and sudden death in animals. In this study, osteomyelitis was diagnosed in small, lame, 11- to 12-wk-old male turkeys. Lameness and slower growth among the turkeys was observed on the farm. During pathological examination, multiple lesions were found in the metaphyseal and physeal areas of the femurs, tibiotarsi, and tarsometatarsi, with multifocal to coalescing mixed heterophilic/granulomatous necrotizing osteomyelitis. Y. pseudotuberculosis was isolated from the femoral and tibiotarsal bones or from the joints of six lame turkeys sent for necropsy. The isolation required homogenizing of lesion tissue in phosphate-mannitol-peptone broth, which was cultured directly - and, if needed, after cold enrichment - on selective cefsulodin-irgasan-novobiocin agar. Whole-genome sequencing was used for identification and typing. All isolates belonged to bio/serotype 1/O:1a and sequence type ST42 (Achtman scheme), which is commonly reported in both human and animal Y. pseudotuberculosis infections in Europe. The isolates from all six turkeys showed only one to two allele differences in the core genome comparison, indicating a common source of infection. All asymptomatic turkeys were slaughtered at the age of 17 weeks. Whole and partial carcass condemnation rates at the slaughterhouse were high, but no macroscopic changes in the skeletal system were found, showing that food chain information is essential. This study confirms earlier findings that Y. pseudotuberculosis can cause osteomyelitis in fattening turkeys, leading to lameness. Food chain information is essential for slaughterhouse operations, to protect the workers and emphasize good working hygiene during slaughter.
Topics: Animals; Lameness, Animal; Male; Osteomyelitis; Turkeys; Yersinia pseudotuberculosis; Yersinia pseudotuberculosis Infections
PubMed: 35429816
DOI: 10.1016/j.vetmic.2022.109424 -
Harnessing β-Lactam Antibiotics for Illumination of the Activity of Penicillin-Binding Proteins in .ACS Chemical Biology May 2020Selective chemical probes enable individual investigation of penicillin-binding proteins (PBPs) and provide critical information about their enzymatic activity with...
Selective chemical probes enable individual investigation of penicillin-binding proteins (PBPs) and provide critical information about their enzymatic activity with spatial and temporal resolution. To identify scaffolds for novel probes to study peptidoglycan biosynthesis in , we evaluated the PBP inhibition profiles of 21 β-lactam antibiotics from different structural subclasses using a fluorescence-based assay. Most compounds readily labeled PBP1, PBP2a, PBP2b, or PBP4. Almost all penicillin scaffolds were coselective for all or combinations of PBP2a, 2b, and 4. Cephalosporins, on the other hand, possessed the lowest IC values for PBP1 alone or along with PBP4 (ceftriaxone, cefoxitin) and 2b (cefotaxime) or 2a, 2b, and 4 (cephalothin). Overall, five selective inhibitors for PBP1 (aztreonam, faropenem, piperacillin, cefuroxime, and cefsulodin), one selective inhibitor for PBP5 (6-aminopenicillanic acid), and various coselective inhibitors for other PBPs in were discovered. Surprisingly, carbapenems strongly inhibited PBP3, formerly shown to have low affinity for β-lactams and speculated to be involved in β-lactam resistance in . To investigate the specific roles of PBP3, we developed activity-based probes based on the meropenem core and utilized them to monitor the activity of PBP3 in living cells. We showed that PBP3 activity localizes as patches in single cells and concentrates as a ring at the septum and the division site during the cell growth cycle. Our activity-based approach enabled spatial resolution of the transpeptidation activity of individual PBPs in this model microorganism, which was not possible with previous chemical and biological approaches.
Topics: Acetylglucosamine; Anti-Bacterial Agents; Bacillus subtilis; Cell Division; Cell Proliferation; Drug Evaluation, Preclinical; Enzyme Activation; Enzyme Inhibitors; Fluorescent Dyes; Glycosylation; Humans; Lighting; Muramic Acids; Optical Imaging; Penicillin-Binding Proteins; Structure-Activity Relationship; beta-Lactams
PubMed: 32155044
DOI: 10.1021/acschembio.9b00977