The use of aminopenicillins in animals within the EU, emergence of resistance in bacteria of animal and human origin and its possible impact on animal and human health.

Aminopenicillins have been widely used for decades for the treatment of various infections in animals and humans in European countries. Following this extensive use, acquired resistance has emerged among human and animal pathogens and commensal bacteria. Aminopenicillins are important first-line treatment options in both humans and animals, but are also among limited therapies for infections with enterococci and Listeria spp. in humans in some settings. Therefore, there is a need to assess the impact of the use of these antimicrobials in animals on public and animal health. The most important mechanisms of resistance to aminopenicillins are the ß-lactamase enzymes. Similar resistance genes have been detected in bacteria of human and animal origin, and molecular studies suggest that transmission of resistant bacteria or resistance genes occurs between animals and humans. Due to the complexity of epidemiology and the near ubiquity of many aminopenicillin resistance determinants, the direction of transfer is difficult to ascertain, except for major zoonotic pathogens. It is therefore challenging to estimate to what extent the use of aminopenicillins in animals could create negative health consequences to humans at the population level. Based on the extent of use of aminopenicillins in humans, it seems probable that the major resistance selection pressure in human pathogens in European countries is due to human consumption. It is evident that veterinary use of these antimicrobials increases the selection pressure towards resistance in animals and loss of efficacy will at minimum jeopardize animal health and welfare.

Human faeces-associated extended-spectrum ß-lactamase-producing Escherichia coli discharge into sanitation systems in 2015 and 2030: a global and regional analysis.

BACKGROUND: Improving management of and treatment within sanitation waste streams could slow the development and transmission of antimicrobial-resistant organisms, but the magnitude of impact has not been quantified. Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli are a major cause of antimicrobial-resistant infections and are frequently detected in faecal waste streams, making them model indicators of the distribution of antimicrobial-resistant organisms that are transmitted through the faecal-oral route. We aimed to estimate the mass of faeces containing ESBL-producing E coli entering different levels of the sanitation ladder globally and by WHO region to determine the global scale at which sanitation infrastructure serves as a vehicle for dissemination of antimicrobial-resistant organisms. METHODS: In this global and regional analysis, we used publicly available sanitation coverage data from the WHO/UNICEF Joint Monitoring Programme and most recent available scientific literature on human faecal production (2018) and carriage of ESBL-producing E coli by healthy individuals (2016) to estimate the quantity of faeces that has been discharged that contains ESBL-producing E coli for 2015 and projected for 2030. We estimated the mass of faeces containing ESBL-producing E coli by WHO region and at different levels of the Sustainable Development Goal sanitation ladder-ie, into at-least basic (ie, safely managed or basic) systems, limited systems, and unimproved systems, and via open defecation. We modelled three scenarios in which the proportion of ESBL-producing E coli among all E coli that was excreted by carriers varied on the basis of the scientific literature: 100% (scenario A), 10% (scenario B), or 1% (scenario C). FINDINGS: Under scenario B, we estimated that approximately 19 billion kg of faeces carrying ESBL-producing E coli was excreted in 2015 globally. Approximately 65·8% (1·2-120 billion kg depending on modelled scenario) of this faecal biomass was managed in at-least basic sanitation systems, 8·4% (160 million-16 billion kg) in limited sanitation systems, 14·4% (270 million-27 billion kg) in unimproved sanitation systems, and 11·4% (220 million-22 billion kg) was openly defecated. The regions with the highest proportion of openly defecated faeces containing ESBL-producing E coli were the South-East Asia (29·4%) and African (21·8%) regions. The South-East Asia, Western Pacific, and African regions produced 524 billion kg (63%) of the total global human faecal biomass, but 16·9 billion kg (90%) of faeces containing ESBL-producing E coli under scenario B. By 2030, estimates under scenario B will have approximately doubled to 37·6 billion kg of faeces carrying ESBL-producing E coli under the most conservative projections. INTERPRETATION: At-least basic sanitation does not guarantee effective removal or inactivation of antimicrobial-resistant organisms from faecal biomass. However, our findings indicate the need for mitigating transport of antimicrobial-resistant organisms via sanitation systems that are not safely managed, including open defecation, which might result in direct environmental discharge and subsequent risk of transmission back to humans. FUNDING: None.

Retrospective analysis of the global antibiotic residues that exceed the predicted no effect concentration for antimicrobial resistance in various environmental matrices.

BACKGROUND: Antimicrobial resistance (AMR) is a growing public health concern. Recent research has suggested that interactions between pathogens and antibiotic residues in various environmental matrices promote the development and spread of AMR in the environment. The levels of antibiotic residues in the aquatic environment have been analysed globally. Recently, Predicted No Effect Environmental Concentration (PNEC) values for many antibiotics have been suggested, based on their estimated minimal selective concentrations for selected bacterial species. The PNEC values can serve as a guide on the maximum levels of antibiotic residues in an environmental matrix, below which resistance is unlikely to develop. AIM: We aimed to determine which of the antibiotics, considered as "priority antibiotics" by the World Health Organisation (WHO), most frequently exceeded their PNEC values in the global aquatic environment. METHODS: We obtained data from the German Environment Agency pharmaceutical database on means, medians or single values of 12 antibiotic types in five different environmental matrices [municipal wastewater treatment plant effluent, industrial wastewater effluent, hospital wastewater effluent, surface water, and drinking water] across 47 countries. We compared the mean levels of the 12 antibiotics in each environmental matrix to their suggested PNEC values to determine which antibiotic types exceeded PNEC and were most likely to select for resistance. We also determined which environmental matrices and countries had the highest burden of antibiotic residues. RESULTS: Our study revealed that 7.9% of all analyses of antibiotic residues performed in the environmental matrices globally exceeded PNEC. Ciprofloxacin and clarithromycin had the greatest proportion (>30%) of residues exceeding PNEC. Hospital wastewater and industrial wastewater had the highest burden of antibiotic residues exceeding PNEC. No antibiotics exceeded PNEC in drinking water. CONCLUSION: While most environmental monitoring studies have focused on municipal wastewater treatment plants, the limited number of studies on hospital wastewater and industrial wastewater revealed that a large number of antibiotic residues coming from these sources exceeded their PNEC values. Our study highlights the importance of implementing on-site treatment systems that aim to destroy antibiotics prior to discharging wastewater to surface waters. Attention needs to be focused on the role that environmental matrices, particularly our wastewater sites, play in promoting antibiotic resistance. Novel treatment technologies need to be developed and implemented to increase the removal efficiencies of treatment plants and from antibiotic manufacturing, and decrease the discharge of antibiotic residues into aquatic environments.

The use of aminoglycosides in animals within the EU: development of resistance in animals and possible impact on human and animal health: a review.

Aminoglycosides (AGs) are important antibacterial agents for the treatment of various infections in humans and animals. Following extensive use of AGs in humans, food-producing animals and companion animals, acquired resistance among human and animal pathogens and commensal bacteria has emerged. Acquired resistance occurs through several mechanisms, but enzymatic inactivation of AGs is the most common one. Resistance genes are often located on mobile genetic elements, facilitating their spread between different bacterial species and between animals and humans. AG resistance has been found in many different bacterial species, including those with zoonotic potential such as Salmonella spp., Campylobacter spp. and livestock-associated MRSA. The highest risk is anticipated from transfer of resistant enterococci or coliforms (Escherichia coli) since infections with these pathogens in humans would potentially be treated with AGs. There is evidence that the use of AGs in human and veterinary medicine is associated with the increased prevalence of resistance. The same resistance genes have been found in isolates from humans and animals. Evaluation of risk factors indicates that the probability of transmission of AG resistance from animals to humans through transfer of zoonotic or commensal foodborne bacteria and/or their mobile genetic elements can be regarded as high, although there are no quantitative data on the actual contribution of animals to AG resistance in human pathogens. Responsible use of AGs is of great importance in order to safeguard their clinical efficacy for human and veterinary medicine.

Marine Bivalve Mollusks As Possible Indicators of Multidrug-Resistant Escherichia coli and Other Species of the Enterobacteriaceae Family.

The mechanisms for the development and spread of antibacterial resistance (ABR) in bacteria residing in environmental compartments, including the marine environment, are far from understood. The objective of this study was to examine the ABR rates in Escherichia coli and other Enterobacteriaceae isolates obtained from marine bivalve mollusks collected along the Norwegian coast during a period from October 2014 to November 2015. A total of 549 bivalve samples were examined by a five times three tube most probable number method for enumeration of E. coli in bivalves resulting in 199 isolates from the positive samples. These isolates were identified by biochemical reactions and matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry, showing that 90% were E. coli, while the remaining were species within the genera Klebsiella, Citrobacter, and Enterobacter. All 199 isolates recovered were susceptibility tested following the European Committee on Antimicrobial Susceptibility Testing disk diffusion method. In total, 75 of 199 (38%) isolates showed resistance to at least one antibacterial agent, while multidrug-resistance were seen in 9 (5%) isolates. One isolate conferred resistance toward 15 antibacterial agents. Among the 75 resistant isolates, resistance toward extended-spectrum penicillins (83%), aminoglycosides (16%), trimethoprim (13%), sulfonamides (11%), tetracyclines (8%), third-generation cephalosporins (7%), amphenicols (5%), nitrofurans (5%), and quinolones (5%), were observed. Whole-genome sequencing on a selection of 10 E. coli isolates identified the genes responsible for resistance, including blaCTX-M genes. To indicate the potential for horizontal gene transfer, conjugation experiments were performed on the same selected isolates. Conjugative transfer of resistance was observed for six of the 10 E. coli isolates. In order to compare E. coli isolates from bivalves with clinical strains, multiple-locus variable number tandem repeats analysis (MLVA) was applied on a selection of 30 resistant E. coli isolates. The MLVA-profiles were associated with community-acquired E. coli strains causing bacteremia. Our study indicates that bivalves represent an important tool for monitoring antibacterial resistant E. coli and other members of the Enterobacteriaceae family in the coastal environment.

Carriage of ESBL/AmpC-producing or ciprofloxacin non-susceptible Escherichia coli and Klebsiella spp. in healthy people in Norway.

BACKGROUND: Asymptomatic carriage has been recognised as an important risk factor for infection caused by antibiotic resistant bacteria. A 14% global prevalence of Extended-Spectrum Beta-lactamase (ESBL) carriage was recently reported, but large intra-and interregional variations were observed. We investigated the faecal carriage rates of ESBL-, AmpC-producing and ciprofloxacin non-susceptible Escherichia coli and Klebsiella spp. in healthy Norwegians. METHODS: Rectal samples were obtained from 284 volunteers, together with demographic data and information on recent travel history. The rectal samples were screened by selective plating and E. coli and Klebsiella spp. identified using MALDI-TOF. Phenotypic and molecular characterization of resistant isolates was also performed. RESULTS: ESBL- or AmpC-producing E. coli and Klebsiella spp. were isolated from 4.9% and 3.2% of the study population, respectively. Carriage of ciprofloxacin non-susceptible isolates was detected in 9.9% of the volunteers. Molecular typing of ESBL/plasmid-mediated AmpC (pAmpC)-producing isolates suggested an allodemic situation rather than the dissemination of a specific clone in the Norwegian community. In concurrence with previous findings, travel to South-East Asia was associated with increased risk of carrying resistant E. coli or Klebsiella spp., highlighting the contribution of factors such as increased global mobility in erasing the boundaries between healthcare and community settings when it comes to spread of resistant bacteria. CONCLUSIONS: Overall, our study recognised Norway as a low-incidence country for faecal carriage of resistant bacteria among healthy individuals. Furthermore, our work denoted the importance of healthy humans as a reservoir for transmission of antibiotic resistant E. coli and Klebsiella spp.

Shiga toxin-producing escherichia coli infections in Norway, 1992-2012: characterization of isolates and identification of risk factors for haemolytic uremic syndrome.

BACKGROUND: Shiga toxin-producing E. coli (STEC) infection is associated with haemolytic uremic syndrome (HUS). Therefore Norway has implemented strict guidelines for prevention and control of STEC infection. However, only a subgroup of STEC leads to HUS. Thus, identification of determinants differentiating high risk STEC (HUS STEC) from low risk STEC (non-HUS STEC) is needed to enable implementation of graded infectious disease response. METHODS: A national study of 333 STEC infections in Norway, including one STEC from each patient or outbreak over two decades (1992-2012), was conducted. Serotype, virulence profile, and genotype of each STEC were determined by phenotypic or PCR based methods. The association between microbiological properties and demographic and clinical data was assessed by univariable analyses and multiple logistic regression models. RESULTS: From 1992 through 2012, an increased number of STEC cases including more domestically acquired infections were notified in Norway. O157 was the most frequent serogroup (33.6 %), although a decrease of this serogroup was seen over the last decade. All 25 HUS patients yielded STEC with stx2, eae, and ehxA. In a multiple logistic regression model, age ≤5 years (OR = 16.7) and stx2a (OR = 30.1) were independently related to increased risk of HUS. eae and hospitalization could not be modelled since all HUS patients showed these traits. The combination of low age (≤5 years) and the presence of stx2a, and eae gave a positive predictive value (PPV) for HUS of 67.5 % and a negative predictive value (NPV) of 99.0 %. SF O157:[H7] and O145:H?, although associated with HUS in the univariable analyses, were not independent risk factors. stx1 (OR = 0.1) was the sole factor independently associated with a reduced risk of HUS (NPV: 79.7 %); stx2c was not so. CONCLUSIONS: Our results indicate that virulence gene profile and patients' age are the major determinants of HUS development.

Evaluation of the ability of four ESBL-screening media to detect ESBL-producing Salmonella and Shigella.

BACKGROUND: The aim of this study was to compare the ability of four commercially available media for screening extended-spectrum beta-lactamase (ESBL) to detect and identify ESBL-producing Salmonella and Shigella in fecal samples. A total of 71 Salmonella- and 21 Shigella-isolates producing ESBL(A) and/or AmpC, were received at Norwegian Institute of Public Health between 2005 and 2012. The 92 isolates were mixed with fecal specimens and tested on four ESBL screening media; ChromID ESBL (BioMèrieux), Brilliance ESBL (Oxoid), BLSE agar (AES Chemunex) and CHROMagar ESBL (CHROMagar). The BLSE agar is a biplate consisting of two different agars. Brilliance and CHROMagar are supposed to suppress growth of AmpC-producing bacteria while ChromID and BLSE agar are intended to detect both ESBL(A) and AmpC. RESULTS: The total sensitivity (ESBL(A)+AmpC) with 95% confidence intervals after 24 hours of incubation were as follows: ChromID: 95% (90.4-99.6), Brilliance: 93% (87.6-98.4), BLSE agar (Drigalski): 99% (96.9-100), BLSE agar (MacConkey): 99% (96.9-100) and CHROMagar: 85% (77.5-92.5). The BLSE agar identified Salmonella and Shigella isolates as lactose-negative. The other agars based on chromogenic technology displayed Salmonella and Shigella flexneri isolates with colorless colonies (as expected). Shigella sonnei produced pink colonies, similar to the morphology described for E. coli. CONCLUSION: All four agar media were reliable in screening fecal samples for ESBL(A)-producing Salmonella and Shigella. However, only ChromID and BLSE agar gave reliable detection of AmpC-producing isolates. Identification of different bacterial species based on colony colour alone was not accurate for any of the four agars.

Age-related differences in symptoms, diagnosis and prognosis of bacteremia.

BACKGROUND: Elderly patients are at particular risk for bacteremia and sepsis. Atypical presentation may complicate the diagnosis. We studied patients with bacteremia, in order to assess possible age-related effects on the clinical presentation and course of severe infections. METHODS: We reviewed the records of 680 patients hospitalized between 1994 and 2004. All patients were diagnosed with bacteremia, 450 caused by Escherichia coli and 230 by Streptococcus pneumoniae. Descriptive analyses were performed for three age groups (< 65 years, 65-84 years, ≥ 85 years). In multivariate analyses age was dichotomized (< 65, ≥ 65 years). Symptoms were categorized into atypical or typical. Prognostic sensitivity of CRP and SIRS in identifying early organ failure was studied at different cut-off values. Outcome variables were organ failure within one day after admission and in-hospital mortality. RESULTS: The higher age-groups more often presented atypical symptoms (p <0.001), decline in general health (p=0.029), and higher in-hospital mortality (p<0.001). The prognostic sensitivity of CRP did not differ between age groups, but in those ≥ 85 years the prognostic sensitivity of two SIRS criteria was lower than that of three criteria. Classical symptoms were protective for early organ failure (OR 0.67, 95% CI 0.45-0.99), and risk factors included; age ≥ 65 years (OR 1.65, 95% CI 1.09-2.49), comorbid illnesses (OR 1.19, 95% CI 1.02-1.40 per diagnosis), decline in general health (OR 2.28, 95% CI 1.58-3.27), tachycardia (OR 1.50, 95% CI 1.02-2.20), tachypnea (OR 3.86, 95% CI 2.64-5.66), and leukopenia (OR 4.16, 95% CI 1.59-10.91). Fever was protective for in-hospital mortality (OR 0.46, 95% CI 0.24-0.89), and risk factors included; age ≥ 65 years (OR 15.02, 95% CI 3.68-61.29), ≥ 1 comorbid illness (OR 2.61, 95% CI 1.11-6.14), bacteremia caused by S. pneumoniae (OR 2.79, 95% CI 1.43-5.46), leukopenia (OR 4.62, 95% CI 1.88-11.37), and number of early failing organs (OR 3.06, 95% CI 2.20-4.27 per failing organ). CONCLUSIONS: Elderly patients with bacteremia more often present with atypical symptoms and reduced general health. The SIRS-criteria have poorer sensitivity for identifying organ failure in these patients. Advanced age, comorbidity, decline in general health, pneumococcal infection, and absence of classical symptoms are markers of a poor prognosis.

Is the concentration of C-reactive protein in bacteraemia associated with age?

BACKGROUND: C-reactive protein (CRP) is an indicator of inflammation, and is often used in the diagnosis of bacterial infections. It is poorly known whether CRP in bacterial infection is age-dependent. METHODS: Adult patients with a positive blood culture with E. coli or S. pneumoniae during 1994-2004 were included. CRP measured on the same date as the blood cultures were drawn (CRP1), 2-3 days (CRP2) and 4-7 days later (CRP3), were retrieved. The patients were divided into three age groups, < 65, 65-84, and > or = 85, respectively. We studied three cut-off values for CRP and produced age-specific receiver operating characteristics (ROC) curves, using patients with acute coronary or cerebral infarction as controls. RESULTS: 890 patients and 421 controls were available. There was a statistically significant negative correlation between age and CRP1 - 0.072 (p = 0.032). The median CRP1 and CRP2 were significantly higher in the youngest age group. The area under the ROC-curve for the youngest age group was significantly greater than that of the two other age groups, but we found no statistically significant differences in sensitivity related to age. The diagnostic sensitivity of CRP was better for S. pneumoniae than for E. coli, 92.6% vs. 88.0% (p = 0.046) for a cut-off value of 40 mg/L, and 82.4% vs. 61.5% (p =< 0.01) for a cut-off value of 120 mg/L. CONCLUSION: CRP is better in identifying infection with S. pneumoniae than with E. coli. We found a weakening of the CRP-response with age, but this is hardly of clinical significance.

New-onset atrial fibrillation in bacteremia is not associated with C-reactive protein, but is an indicator of increased mortality during hospitalization.

BACKGROUND: Several studies have associated elevated C-reactive protein (CRP) levels to the occurrence of atrial fibrillation (AF). We sought to estimate the frequency and prognostic impact of AF in patients with bacteremia, and to study the possible association between AF and CRP as well as between AF and mortality in this population. METHODS: We retrospectively evaluated patient charts of patients with bacteremia with Escherichia coli or Streptococcus pneumoniae admitted to the Aker University Hospital in Oslo between 1994 and 2004. Known cardiac risk factors for AF, signs and mode of conversion of AF, and, if applicable, date of death were registered, as were characteristics of infection, such as systemic inflammatory response syndrome and white blood cell count. Initial CRP values were categorized into 4 strata. Odds ratios of the 3 highest CRP categories compared with the lowest were obtained from logistic models adjusting for known cardiac risk factors for AF as well as possible factors that may have had an impact on the odds ratios for the different CRP levels. Cox regression analysis was used to compare new-onset AF and death during the first 2 weeks after hospitalization. RESULTS: A total of 672 patient charts were studied; 104 patients (15.4%) had new-onset AF. Peak incidence of new-onset AF occurred on the day of admission. Peak CRP values were reached during the following 2 days. High CRP level at admission did not predict the occurrence of AF. The observed mortality was higher among patients with new-onset AF (p = 0.001) during the first 2 weeks after hospitalization, but this effect disappears when adjusted for relevant factors. CONCLUSIONS: The frequency of new-onset AF in bacteremia is substantial. Initial CRP levels or white blood cell count do not seem to predict new-onset AF, as opposed to systemic inflammatory response syndrome. On the other hand, in patients with bacteremia, new-onset AF should be viewed as an indicator of increased mortality and morbidity.