Cefotaxime-resistant Bacteria Research Articles

Transmission of human-pet antibiotic resistance via aerosols in pet hospitals of Changchun

In recent years, aerosols have been recognized as a prominent medium for the transmission of antibiotic-resistant bacteria and genes. Among these, particles with a particle size of 2 μm (PM2.5) can directly penetrate the alveoli. However, the presence of antibiotic-resistant genes in aerosols from pet hospitals and the potential risks posed by antibiotic-resistant bacteria in these aerosols to humans and animals need to be investigated. In this study, cefotaxime-resistant bacteria were collected from 5 representative pet hospitals in Changchun using a Six-Stage Andersen Cascade Impactor. The distribution of bacteria in each stage was analyzed, and bacteria from stage 5 and 6 were isolated and identified. Minimal inhibitory concentrations of isolates against 12 antimicrobials were determined using broth microdilution method. Quantitative Polymerase Chain Reaction was employed to detect resistance genes and mobile genetic elements that could facilitate resistance spread. The results indicated that ARBs were enriched in stage 5 (1.1–2.1 μm) and stage 3 (3.3–4.7 μm) of the sampler. A total of 159 isolates were collected from stage 5 and 6. Among these isolates, the genera Enterococcus spp. (51%), Staphylococcus spp. (19%), and Bacillus spp. (14%) were the most prevalent. The isolates exhibited the highest resistance to tetracycline and the lowest resistance to cefquinome. Furthermore, 56 (73%) isolates were multidrug-resistant. Quantitative PCR revealed the expression of 165 genes in these isolates, with mobile genetic elements showing the highest expression levels. In conclusion, PM2.5 from pet hospitals harbor a significant number of antibiotic-resistant bacteria and carry mobile genetic elements, posing a potential risk for alveolar infections and the dissemination of antibiotic resistance genes.

Spatial Distribution and Seasonal Variation of Antibiotic-Resistant Bacteria in an Urban River in Northeast China

As the largest freshwater river flowing through Harbin, the Songhua River is a standby water source. It is very important to know the species and distribution of antibiotic-resistant bacteria (ARB) in the river. In this study, five antibiotics were selected to screen and identify ARB in spring and autumn. The results showed that the concentration of cefotaxime-resistant bacteria was the highest, and the maximum concentration at S6 in spring was up to 1.40 × 104 CFU/mL. In spring and autumn, bacteria resistant to three antibiotics were screened at S1 of the Songhua River, and bacteria resistant to five antibiotics were screened at S6. No multiple antibiotic-resistant bacteria (MARB) were screened in the other four sites in autumn, while MARB were screened in the other three samples except S2 in spring. In all sample areas in spring and autumn, the probability of screening MARB at S1 and S6 was the highest, reaching 100%. The identification results of 16S rDNA polymerase chain reaction (PCR) products of ARB showed that a total of 51 ARB strains from 15 bacterial genera were screened in the Songhua River, of which 20 ARB strains were from Pseudomonas. Among the 15 bacterial genera, bacteria from 8 bacterial genera have pathogenicity. The results of this study revealed the concentration, spatial distribution, and seasonal variation of culturable ARB in the Songhua River, providing data support for the remediation of antibiotic resistance gene (ARG) pollution in the river.

Swine-manure composts induce the enrichment of antibiotic-resistant bacteria but not antibiotic resistance genes in soils

Composting is a common and effective strategy for reducing antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) from animal manure. However, it is unclear whether the advantages of composting for the control of ARGs and ARB can be further increased in land application. This study investigated the fate of ARB and ARGs after land application of swine-manure composts (SMCs) to three different soil types (red soil, loess and black soil). The results showed that although the SMCs caused an increase in the abundance of total ARGs in the soil in the short period, they significantly reduced (p < 0.01) the abundance of total ARGs after 82 days compared to the control. The decay rate of ARGs reflected by the half-life times (t1/2) varied by soil type, with red soil being the longest. The SMCs mainly introduced ermF, tetG and tetX into the soils, while these ARGs quickly declined to the control level. Notably, SMCs increased the number of ARB in the soils, especially for cefotaxime-resistant bacteria. Although SMCs only affected the microbiome significantly during the early stage (p < 0.05), it took a much longer time for the microbiome to recover compared to the control. Statistical analysis indicated that changes in the microbial community contributed more to the fate of ARGs during SMCs land application than other factors. Overall, it is proposed that the advantages of ARGs control in the composting process for swine manure can be further increased in land application, but it can still bring some risks in regard to ARB.

Microplastics accumulate priority antibiotic-resistant pathogens: Evidence from the riverine plastisphere

Microplastics (MPs) might accumulate and transport antibiotic-resistant bacteria (ARB) in aquatic systems. We determined the abundance and diversity of culturable ciprofloxacin- and cefotaxime-resistant bacteria in biofilms covering MPs placed in river water, and characterized priority pathogens from these biofilms. Our results showed that the abundance of ARB colonizing MPs tends to be higher compared to sand particles. Also, higher numbers were cultivated from a mixture of polypropylene (PP), polyethylene (PE) and polyethylene terephthalate (PET), compared to PP and PET alone. Aeromonas and Pseudomonas isolates were the most frequently retrieved from MPs placed before a WWTP discharge while Enterobacteriaceae dominated the culturable plastisphere 200 m after the WWTP discharge. Ciprofloxacin- and/or cefotaxime-resistant Enterobacteriaceae (n = 54 unique isolates) were identified as Escherichia coli (n = 37), Klebsiella pneumoniae (n = 3), Citrobacter spp. (n = 9), Enterobacter spp. (n = 4) and Shigella sp. (n = 1). All isolates presented at least one of the virulence features tested (i.e. biofilm formation, haemolytic activity and production of siderophores), 70% carried the intI1 gene and 85% exhibited a multi-drug resistance phenotype. Plasmid-mediated quinolone resistance genes were detected in ciprofloxacin-resistant Enterobacteriaceae [aacA4-cr (40% of the isolates), qnrS (30%), qnrB (25%), and qnrVC (8%)], along with mutations in gyrA (70%) and parC (72%). Cefotaxime-resistant strains (n = 23) harbored blaCTX-M (70%), blaTEM (61%) and blaSHV (39%). Among CTX-M producers, high-risk clones of E. coli (e.g. ST10 or ST131) and K. pneumoniae (ST17) were identified, most of which carrying blaCTX-M-15. Ten out of 16 CTX-M producers were able to transfer blaCTX-M to a recipient strain. Our results demonstrated the occurrence of multidrug resistant Enterobacteriaceae in the riverine plastisphere, harboring ARGs of clinical concern and exhibiting virulence traits, suggesting a contribution of MPs to the dissemination of antibiotic-resistant priority pathogens. The type of MPs and especially water contamination (e.g. by WWTPs discharges) seem to determine the resistome of the riverine plastisphere.

Field emission scanning electron microscopic, X-ray diffraction and ultraviolet spectroscopic analysis of Terminalia bellerica based silver nanoparticles and evaluation of their antioxidant, catalytic and antibacterial activity

In recent years, scientists have come up with ways to make nanoparticles that are inexpensive and good for the environment. Terminalia bellerica-based silver nanoparticles (TBAgNPs) were made in this study using methanol extract from T. bellerica fruits. This method was quick, economical, and good for the environment. The biosynthesized TBAgNPs were used as antioxidants, antibacterial agents, and anti-catalytic agents. Analytical techniques like XRD, FESEM, and UV–Vis were used to find out more about the spherical TBAgNPs that were made. Also, Cefotaxime-resistant bacteria found in hospitals were used to test how well the TBAgNPs killed bacteria. With the Bauer-agar Kirby's gel diffusion and Mueller-Hinton broth methods, the ability of the synthesized TBAgNPs to stop bacterial growth was tested. After the TBAgNPs were studied, it was found that the average size of their crystals was between 10 and 25 nm. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) reducing tests showed that these AgNPs could act as antioxidants, and TBAgNPs (%inhibition = 20.90% to 94.94%) were better antioxidant than ascorbic acid (%inhibition = 13.80% to 86.10%) and extract (%inhibition = 16.90% to 80.50%). The reduction of methylene blue (MB) to leucomethylene blue (LMB) with sodium borohydride (NaBH4) was used as a model to test the catalytic potential of TBAgNPs. On UV spectroscopic analysis at room temperature, TBAgNPs at different concentrations were able to reduce methylene blue effectively. For Escherichia coli and Klebsiella pneumoniae, the minimum inhibitory concentration (MIC) for TBAgNPs was 0.625 μg/mL and 1.25 μg/mL, respectively. Based on these results, silver nanoparticles made with Terminalia bellerica extract may have much biological importance and could be used in making useful therapeutic applications.

Dumpsite leachate as a hotspot of multidrug resistant Enterobacteriaceae harbouring extended spectrum and AmpC β-lactamase genes; a case study of Awotan municipal solid waste dumpsite in Southwest Nigeria

Leachates generated by solid waste dumpsites could be a factor in the development of antibiotic resistance in environmental bacteria. In Nigeria, there is a dearth of information on the contribution of leachates from dumpsite to the ever-increasing trend of antibiotic resistance. A total of 250 cefotaxime-resistant bacteria recovered from dumpsite leachate was screened for extended spectrum β-lactamase (ESBL) production using the double disc synergy test and 41 were ESBL producers. The ESBL producers belonged to four genera and were identified as Escherichia (20), Citrobacter (13), Enterobacter (6) and Raoultella (2). Antibiotic susceptibility testing was carried out using disk diffusion method, while detection of extended spectrum and AmpC β-lactamase genes was done using Real-Time PCR. All the ESBL- producing bacteria were multidrug resistant, with 100% resistance observed to cefotaxime, cefpodoxime, tetracycline, sulfonamide, ciprofloxacin, ampicillin, ertapenem and imipenem, while 81.6% and 63.1% of the ESBL producers showed resistance to ceftazidime and amoxicillin-clavulanate respectively. blaCTX-M-1 and blaCTX-M-9 groups of CTX-M β-lactamases were detected in 33 (80.5%) of the isolates, while 26 (63.4%) harboured the blaCTX-M-2, blaCTX-M-8 and blaCTX-M-25 groups. blaTEM was detected in 31 (75.6%) of the ESBL producers, while blaSHV was detected in 13 (31.7%). One isolate (E. coli AW-32) carried blaCMY-1 gene. None of the isolates carried blaKPC, blaNDM-1 and blaampC genes, while seven isolates were devoid of any gene. There was co-occurrence of resistance genes in thirty of the isolates. This study has revealed that dumpsite leachates could be important reservoirs of multidrug resistant Enterobacteriaceae harbouring extended spectrum and AmpC β-lactamases genes.

Selection of antibiotic resistance by metals in a riverine bacterial community

Antibiotic resistance is a health challenge across human, animal and environmental settings. In the environment, metals may contribute to antibiotic resistance selection. This study aimed to investigate the role of copper and zinc in the selection of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in a riverine bacterial community. Using a microcosm approach, bacteria in water samples were exposed to 50 μg L−1 and 100 μg L−1 of copper and zinc, for 20 days. The prevalence of ARB was determined from colony forming units counts in media with and without antibiotics. A significant increase in the prevalence of cefotaxime-resistant (from 2.3% in control to 9.5% in Cu50 and 16.8% in Cu100) and tetracycline-resistant bacteria (from 0.03% to 0.23% in Cu100) was observed in communities exposed to copper. Zinc exposure resulted in an increase in the prevalence of cefotaxime-resistant bacteria (from 24.6% to 91.3% in Zn50 and 72.4% in Zn100) and of kanamycin-resistant bacteria (from 6.1% to 24.1% in Zn50 and 43% in Zn100). Cefotaxime- and kanamycin-resistant bacteria belonged to genera intrinsically resistant to these compounds. DGGE profiling confirmed that metal exposure altered the structure and diversity of bacterial communities. Changes in the abundance of genes usually associated with mobile genetic elements (blaCTX-M, blaTEM, tet(A) and intI1) were not detected after exposure. Results demonstrated the selection of bacteria intrinsically resistant to antibiotics imposed by copper and zinc exposure, suggesting an important role played by cross-resistance mechanisms.

Effect of solar photo-Fenton process in raceway pond reactors at neutral pH on antibiotic resistance determinants in secondary treated urban wastewater.

Solar photo-Fenton process in raceway pond reactors was investigated at neutral pH as a sustainable tertiary treatment of real urban wastewater. In particular, the effect on antibiotic resistance determinants was evaluated. An effective inactivation of different wild bacterial populations was achieved considering total and cefotaxime resistant bacteria. The detection limit (1 CFU mL-1) was achieved in the range 80-100 min (5.4-6.7 kJ L-1 of cumulative solar energy required) for Total Coliforms (TC) (40-60 min for resistant TC, 4.3-5.2 kJ L-1), 60-80 min (4.5-5.4 kJ L-1) for Escherichia coli (E. coli) (40 min for resistant E. coli, 4.1-4.7 kJ L-1) and 40-60 min (3.9-4.5 kJ L-1) for Enterococcus sp. (Entero) (30-40 min for resistant Entero, 3.2-3.8 kJ L-1) with 20 mg L-1 Fe2+ and 50 mg L-1 H2O2. Under these mild oxidation conditions, 7 out of the 10 detected antibiotics were effectively removed (60-100%). As the removal of antibiotic resistance genes (ARGs) is of concern, no conclusive results were obtained, as sulfonamide resistance gene was reduced to some extent (relative abundance <1), meanwhile class 1 integron intI1 and ß-lactam resistance genes were not affected. Accordingly, more research and likely more intensive oxidative conditions are needed for an efficient ARGs removal.

Kinetic assessment of antibiotic resistant bacteria inactivation by solar photo-Fenton in batch and continuous flow mode for wastewater reuse

The presence of antibiotic resistant bacteria in municipal wastewater treatment plants represents a real risk to human health. For the first time, this paper shows that the inactivation rate of cefotaxime resistant bacteria is the same as total bacteria when secondary effluents are treated by the solar photo-Fenton process. To obtain this result, an exhaustive and comparative kinetic study on the inactivation of both total and cefotaxime resistant bacteria (Total coliform, Escherichia coli and Enterococcus sp) was carried out, taking into account the effects of the main operation conditions, such as solar irradiance and iron concentration, and operation mode (batch and continuous). In all the operation conditions studied, no significant differences were found between the first order inactivation rate constants, ki, of total and cefotaxime resistant bacteria. Additionally, ki increased with solar irradiance and iron concentration. As for the effect of the operation mode, the main finding of this work is much quicker inactivation in continuous flow mode than in batch mode, pointing out its potential application at large scale. The best continuous operation condition to inactivate the bacteria to the detection limit (1 CFU mL−1), was at 22.4 min of hydraulic residence time with 5 mg Fe2+ L−1 and 30 mg H2O2·L−1. This treatment time is approximately a third of that reported in batch mode. The efficiency, in terms of figure of merits, of the continuous flow operation was 2.7 m2 of solar collector area to reduce one log of E. coli concentration per m3 of treated water and per hour, in comparison with 2137 m2 calculated for batch operation under the same solar UVA irradiance, 30 W m−2. This paper encourages research into continuous solar disinfection processes due to its enhanced efficiency with regard to the commonly used batch wise operation and shows that efficient removal of total bacteria ensures the removal of antibiotic resistant bacteria.

OCCURRENCE AND MOLECULAR CHARACTERIZATION OF EXTENDED SPECTRUM BETA-LACTAMASE PRODUCING Enterobacteriaceae IN MILK AND SOME DAIRY PRODUCTS

In the present study a total of one hundred samples were randomly purchased from different supermarkets in Damietta city, Egypt, including 25 samples each of raw milk, ice cream, Kareish and Domiati cheese. The collected samples were screened initially for cefotaxime-resistant bacteria using selective enrichment. The suspected colonies were further characterized by analysis of resistance genes in ESBL-producing isolates. A total of 59 ESBLs producers colonies (24 from raw milk, 15 from kareish cheese, 13 from Domiati cheese and 7 from ice cream) were recovered and identified as Escherichia coli (n = 29), Klebsiella pneumonia (n = 8), Klebsiella oxytoca (n = 5), Enterobacter aerogenes (n =8), Citrobacter diversus (n = 8) and Serratia liquefaciens (n = 1). Resistance to two or more antibiotics was observed among the recovered isolates. E. coli isolates showed high resistance pattern against cefaclor (100%), cefoxitin (100%), cefalaxim (96.5%), ceftazidime (93%), pencillin (93%) and 79% for Pipercillin/Tazobactam. All the recovered Klebseilla spp. isolates were resistant to Cefepime and Pipercillin/Tazobactam (84.6% each) and exhibited 100% resistant to other antimicrobials agents. Meanwhile, all the recovered isolates were sensitive to imipenem and meropenem. Gene encoding blaCTX-M1 was mostly predominant among screened genes, being present in 13 (48%) dairy samples for blaTEM and blaSHV, they were detected in 12 (44%) and 4 (14.8%) isolates, respectively. In conclusion, there is clear evidence of circulation of antibiotic-resistant food borne ESBL producing Enterobacteriaceae in the examined dairy samples. The concern about increasing the risk of dissemination of such multi-drug resistant pathogens rises with a potential asymptomatic colonization and complication of systemic infection in human subjects. There is also a possible interface for the exchange of resistance genes within and across species and with commensal bacteria of the human and animals.Key words: dairy products; ESBL producing Enterobacteriaceae; Egypt; public health

High Prevalence of Cefotaxime Resistant Bacteria in Grazing Beef Cattle: A Cross Sectional Study.

Although the over-use of antibiotics during food animal production is a potential driver of antimicrobial resistant microorganisms (ARMs), a high prevalence of cefotaxime resistant bacteria (CRB) has been observed in grazing animals raised without antibiotic supplementation. In this cross-sectional study, the prevalence and concentration of CRB in beef cattle on grazing farms were investigated. Fecal samples from the recto-anal junction of cattle (n = 840) and environmental samples (n = 258) were collected from 17 farms in North and Central Florida in the United States, and a survey of farm characteristics, animal husbandry practices, and antibiotic usage was conducted. CRB were detected in fecal samples from 47.4% of all cattle, with the prevalence ranging from 21.1 to 87.5% on farms, and significantly higher (P < 0.001) in calves compared to adult cows (54.1 vs. 41.8%). Environmental samples had a higher prevalence than fecal samples (P < 0.001), with CRB detected in 88.6% of water, 98.7% of soil, and 95.7% of forage samples. Compared to the concentration (log CFU/g) of CRB in fecal samples (2.95, 95% CI: 2.89, 3.02), the concentration of CRB was higher (P < 0.001) in soil and forage samples (5.37, 95% CI: 5.16, 5.57) and lower (P < 0.001) in water samples (1.08, 95% CI: 0.82, 1.36). Soil microbiota from farms with high prevalence of CRB clustered closer together and the proportion of Phylum Proteobacteria was higher on farms with high prevalence of CRB resistance. Large farming operations were associated with a 58% higher likelihood of CRB detection in fecal samples. Regular cleaning of drinking troughs and the addition of ionophores to feed were associated with CRB reduction in fecal samples. Taken together, the widespread of CRB into both cattle seldom treated with cephalosporin antibiotics and the surrounding environment suggests the environment is a natural source of antimicrobial resistance in beef cattle.

Impact of Vegetative Treatment Systems on Multiple Measures of Antibiotic Resistance in Agricultural Wastewater.

Wastewater is an important vector of antibiotic resistant bacteria and antibiotic resistance genes (ARB/G). While there is broad agreement that ARB/G from agricultural (ag) wastewaters can be transported through the environment and may contribute to untreatable infectious disease in humans and animals, there remain large knowledge gaps surrounding applied details on the types and amounts of ARB/G associated with different agricultural wastewater treatment options and different ag production systems. This study evaluates a vegetative treatment system (VTS) built to treat the wastewater from a beef cattle feedlot. Samples were collected for three years, and plated on multiple media types to enumerate tetracycline and cefotaxime-resistant bacteria. Enterobacteriaceae isolates (n = 822) were characterized for carriage of tetracycline resistance genes, and E. coli isolates (n = 673) were phenotyped to determine multi-drug resistance (MDR) profiles. Tetracycline resistance in feedlot runoff wastewater was 2-to-3 orders of magnitude higher compared to rainfall runoff from the VTS fields, indicating efficacy of the VTA for reducing ARB over time following wastewater application. Clear differences in MDR profiles were observed based on the specific media on which a sample was plated. This result highlights the importance of method, especially in the context of isolate-based surveillance and monitoring of ARB in agricultural wastewaters.

Colonization Dynamics of Cefotaxime Resistant Bacteria in Beef Cattle Raised Without Cephalosporin Antibiotics

The emergence of infections caused by antimicrobial resistant microorganisms (ARMs) is currently one of the most important challenges to public health and medicine. Though speculated to originate at least partially from the overuse of antibiotics during food animal production, we hypothesized that cattle are exposed to ARMs in the environment. In this cohort study, a herd of beef calves with no previous exposure to antibiotics was followed during the first year of life in order to investigate the rate of colonization by bacteria resistant to the third-generation cephalosporin cefotaxime. Fecal samples were collected from the recto anal junction of cattle at the age of ~3, 6, 9, and 12 months and tested for cefotaxime resistant bacteria (CRB) and the presence of extended spectrum β-lactamases (ESBLs). The colonization dynamics of CRB in calves (n = 188) was evaluated with samples collected from four periods using longitudinal statistical analyses. Colonization by CRB was a dynamic process with over 92% of the calves testing positive for CRB at least once during the first year of life. All isolates subjected to antimicrobial susceptibility test were resistant to at least four different antibiotics and carried multiple variants of the blaCTX-M genes. Metagenomic analysis revealed significant differences in microbiota of the calves with and without CRB colonization at different ages. This study provides evidence that colonization of beef calves by ARMs is a dynamic process that can occur in the absence of veterinary or agricultural use of antibiotics.

Studies on drug release kinetics and antibacterial activity against drug-resistant bacteria of cefotaxime sodium loaded layered double hydroxide–fenugreek nanohybrid

Antibacterial activity of a CLF nanohybrid against E. coli 949 ESBL cefotaxime-resistant bacteria via the interaction of penicillin binding protein.

High abundance and diversity of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in faeces and tonsils of pigs at slaughter

This cross-sectional study investigates the abundance of cefotaxime-resistant Escherichia coli (CREC) in the faeces and tonsils of 96 pigs during slaughter. Moreover, different isolates from a selected number of pigs were tested to study the diversity of blaESBL genes within E. coli isolates from one pig. Cefotaxime-resistant bacteria (based on enumeration results on MacConkey agar supplemented with 1mg/L cefotaxime) were found in the faeces of 77 pigs (80%; 95% CI: 70–87%) and the tonsils of 91 pigs (95%; 95% CI: 88%–98%). Cefotaxime-resistant E. coli (based on enumeration results on Tryptone Bile X-glucuronide agar supplemented with 1mg/L cefotaxime) were detected in 72 faecal samples (75%; 95% CI: 64–83%) and 45 tonsil samples (47%; 95% CI: 35–59%), in numbers up to 5.5 and 5.6log10 CFU/g, respectively. On average, around 1/10,000 E. coli in both faeces and tonsils were cefotaxime-resistant, though large variations were observed between pigs. Within one sample, CREC isolates with up to five different combinations of ESBL genes were observed. In three out of 16 faecal samples and six out of 14 tonsil samples, only one ESBL gene profile was found. The high numbers of CREC that are occasionally found in the faeces and tonsils of pigs during slaughter may represent an important source of contamination of carcasses and subsequently pork.

Identification and Characterization of Cefotaxime Resistant Bacteria in Beef Cattle.

Third-generation cephalosporins are an important class of antibiotics that are widely used in treatment of serious Gram-negative bacterial infections. In this study, we report the isolation of bacteria resistant to the third-generation cephalosporin cefotaxime from cattle with no previous cefotaxime antibiotic exposure. The prevalence of cefotaxime-resistant bacteria was examined by a combination of culture based and molecular typing methods in beef cattle (n = 1341) from 8 herds located in North Central Florida. The overall prevalence of cefotaxime-resistant bacteria was 15.8% (95% CI: 13.9, 17.8), varied between farms, and ranged from 5.2% to 100%. A subset of isolates (n = 23) was further characterized for the cefotaxime minimum inhibitory concentration (MIC) and antibiotic susceptibility against 10 different antibiotics, sequencing of nine β- lactamase genes, and species identification by 16S rRNA sequencing. Most of the bacterial isolates were resistant to cefotaxime (concentrations, > 64 μg/mL) and showed high levels of multi-drug resistance. Full length 16S rRNA sequences (~1300 bp) revealed that most of the isolates were not primary human or animal pathogens; rather were more typical of commensal, soil, or other environmental origin. Six extended spectrum β-lactamase (ESBL) genes identical to those in clinical human isolates were identified. Our study highlights the potential for carriage of cefotaxime resistance (including “human” ESBL genes) by the bacterial flora of food animals with no history of cefotaxime antibiotic exposure. A better understanding of the origin and transmission of resistance genes in these pre-harvest settings will be critical to development of strategies to prevent the spread of antimicrobial resistant microorganisms to hospitals and communities.

Extended-spectrum β-lactamase producing Enterobacteriaceae in bulk tank milk from German dairy farms

Although the dairy farm environment is a known source of extended-spectrum β-lactamase (ESBL)-producing bacteria, surveillance data on ESBL in the milk production chain are still scarce. This study aimed at estimating the dimensions of the problem for public health and animal welfare by surveying ESBL-producing Enterobacteriaceae in raw bulk tank milk in Germany. Samples from 866 dairy farms, comprising about 1% of the total number of dairy farms in Germany, were first screened for presence of cefotaxime-resistant bacteria by selective enrichment. Suspect colonies were identified phenotypically and further characterized by biochemical and molecular methods, including analysis of resistance genes and clonal diversity in ESBL-producing isolates. Bulk tank milk from 82 (9.5%) farms yielded Enterobacteriaceae with confirmed ESBL-production. The most frequent ESBL-producing species was Escherichia coli (75.6%), followed by Citrobacter spp. (9.6%), Enterobacter cloacae (6.1%), and Klebsiella oxytoca (3.7%), a few isolates belonged to other species within the genera Hafnia, Raoutella and Serratia. The majority of isolates (95.1%) harbored the β-lactamase blaCTX-M gene, which has gained increased importance among ESBL-producing strains worldwide; the CTX-M group 1 was found to be the dominating (88.4%) phylogenetic group. All ESBL-positive Escherichia coli isolates were clonally heterogeneous, as determined by pulsed-field gel electrophoresis. The results from this survey demonstrate that ESBL-producing bacteria are distributed widely in the dairy farm environment in Germany. Therefore, raw milk is a potential source of exposure for the consumer, which is of increasing importance considering the trend of farmer-to-consumer direct marketing. Furthermore, dairy farm staff have an increased likelihood of exposure to ESBL-producing bacteria. Finally, ESBL-producing bacteria may also be transferred via waste milk to calves, thus further spreading antibiotic resistance in the farm environment.

Assessment of Selected Antibiotic Resistances in Ungrazed Native Nebraska Prairie Soils.

The inherent spatial heterogeneity and complexity of antibiotic-resistant bacteria and antibiotic resistance (AR) genes in manure-affected soils makes it difficult to sort out resistance that can be attributed to human antibiotic use from resistance that occurs naturally in the soil. This study characterizes native Nebraska prairie soils that have not been affected by human or food-animal waste products to provide data on background levels of resistance in southeastern Nebraskan soils. Soil samples were collected from 20 sites enumerated on tetracycline and cefotaxime media; screened for tetracycline-, sulfonamide-, β-lactamase-, and macrolide-resistance genes; and characterized for soil physical and chemical parameters. All prairies contained tetracycline- and cefotaxime-resistant bacteria, and 48% of isolates collected were resistant to two or more antibiotics. Most (98%) of the soil samples and all 20 prairies had at least one tetracycline gene. Most frequently detected were (D), (A) (O), (L), and (B). Sulfonamide genes, which are considered a marker of human or animal activity, were detected in 91% of the samples, despite the lack of human inputs at these sites. No correlations were found between either phenotypic or genotypic resistance and soil physical or chemical parameters. Heterogeneity was observed in AR within and between prairies. Therefore, multiple samples are necessary to overcome heterogeneity and to accurately assess AR. Conclusions regarding AR depend on the gene target measured. To determine the impacts of food-animal antibiotic use on resistance, it is essential that background and/or baseline levels be considered, and where appropriate subtracted out, when evaluating AR in agroecosystems.

High prevalence of the plasmid-mediated quinolone resistance determinant qnrA in multidrug-resistant Enterobacteriaceae from blood cultures in Liverpool, UK

To determine the prevalence of the plasmid-mediated quinolone resistance qnrA gene in a selected collection of blood culture isolates of Enterobacteriaceae resistant to both ciprofloxacin and cefotaxime. Over a 29 month period, a total of 47 non-repetitive isolates of Enterobacteriaceae resistant to both ciprofloxacin and cefotaxime were identified. Isolates were screened for the presence of the qnrA gene, class I integrons and bla(ESBL) by PCR. Transferability was examined by conjugation with the sodium azide-resistant Escherichia coli J53. All qnrA-positive isolates were examined for DNA-relatedness by PFGE. A total of 15 of the 47 test isolates (32%) were positive for the qnrA gene, and included single isolates of E. coli and Citrobacter freundii, 4 Klebsiella pneumoniae and 9 Enterobacter cloacae. All 15 qnrA-positive isolates carried class 1 integrons, and 11 the extended-spectrum beta-lactamase gene bla(SHV-12). By PFGE two K. pneumoniae and three E. cloacae, respectively, were considered clonally but not temporally related. Plasmid transfer of quinolone resistance was only achieved with single isolates of K. pneumoniae and E. cloacae. Both plasmids carried class 1 integrons with a pSAL-1-like gene cassette arrangement intl1-aadA2-qacEDelta-sul1. In this selected group of ciprofloxacin- and cefotaxime-resistant bacteria, carriage of the qnrA gene was high (32%). This compares with <2.0% as demonstrated in worldwide studies of laboratory collections of ciprofloxacin-resistant bacteria. The majority of qnrA-positive isolates in our study originated from high-dependency care units within our hospital, but were shown not to be clonal by PFGE. This is the first report of qnrA-positive Enterobacteriaceae in the United Kingdom.

Cefotaxime therapy of serious infections with multiresistant gram-negative bacilli.

33 patients with serious gram-negative bacillary infections were treated with cefotaxime. In patients with normal renal function the dose varied between 1.5 to 4 g/day. 17 patients had urinary tract infections, 5 respiratory tract infections, 1 combined urinary tract infection and respiratory tract infection, and 10 miscellaneous infections. 16 patients had septicemia. 25 infections were due to pathogens resistant in vitro to ampicillin, cephalothin, gentamicin and/or tobramycin. 15 infections had failed to respond to ampicillin, cefazolin, gentamicin or tobramycin therapy. 32/33 patients responded favourably to cefotaxime (cure or improvement) but 4 patients developed superinfection with cefotaxime-resistant bacteria. No evidence of nephrotoxicity was observed except for a transient moderate rise in creatinine in one patient.