Suchergebnisse

Vancomycin-resistant enterococci (VRE) poses a formidable challenge to public health due to its inherent resistance to multiple antibiotics coupled with the ability to transfer genetic determinants to dangerous pathogens like Methicillin-resistant Staphylococcus aureus (MRSA). The purpose of this study was to investigate the incidence of vancomycin resistance in enterococci among clinical isolates at a tertiary care military hospital in the eastern region of Saudi Arabia and to detect van genes using multiplex-PCR. Overall, 246 isolates of enterococci were collected from various clinical specimens. The isolates were identified, and antimicrobial susceptibility testing was done using the Vitek 2 system. Multiplex PCR was performed on the VRE isolates, thus identified to determine the van genes harbored. A total of 15 VRE were identified, of which 14 (93.3%) were Enterococcus faecium, and 1(6.7%) was Enterococcus casseliflavus with intrinsic vanC resistance. Of the 14 vancomycin-resistant Enterococcus faecium, 8 (57.1%) harbored vanB genes, while 6 (42.8%) harbored vanA genes. All the VRE were susceptible to linezolid and tigecycline. Our study detected a low prevalence (6.1%) of VRE among clinical isolates of enterococci and that the vanB gene predominates in such strains. Susceptibility profiles indicated that linezolid and tigecycline are still effective against these multidrug-resistant pathogens. Pus specimens yielded the highest percentage (53.3%) of isolates from which VRE was obtained, and this finding is novel among studies done in Saudi Arabia.

VRE isolates from pigs (n = 29) and healthy persons (n = 12) recovered during wide surveillance studies performed in Portugal, Denmark, Spain, Switzerland, and the United States (1995 to 2008) were compared with outbreak/prevalent VRE clinical strains (n = 190; 23 countries; 1986 to 2009). Thirty clonally related Enterococcus faecium clonal complex 5 (CC5) isolates (17 sequence type 6 [ST6], 6 ST5, 5 ST185, 1 ST147, and 1 ST493) were obtained from feces of swine and healthy humans. This collection included isolates widespread among pigs of European Union (EU) countries since the mid-1990s. Each ST comprised isolates showing similar pulsed-field gel electrophoresis (PFGE) patterns (≤6 bands difference; >82% similarity). Some CC5 PFGE subtype strains from swine were indistinguishable from hospital vancomycin-resistant enterococci (VRE) causing infections. A truncated variant of Tn1546 (encoding resistance to vancomycin) and tcrB (coding for resistance to copper) were consistently located on 150- to 190-kb plasmids (reppLG1). E. faecium CC17 (ST132) isolates from pig manure and two clinical samples showed identical PFGE profiles and contained a 60-kb mosaic plasmid (repInc18 plus reppRUM) carrying diverse Tn1546-IS1216 variants. The only Enterococcus faecalis isolate obtained from pigs (CC2-ST6) corresponded to a multidrug-resistant clone widely disseminated in hospitals in Italy, Portugal, and Spain, and both animal and human isolates harbored an indistinguishable 100-kb mosaic plasmid (reppRE25 plus reppCF10) containing the whole Tn1546 backbone. The results indicate a current intra- and international spread of E. faecium and E. faecalis clones and their plasmids among swine and humans.

AbstractManure from food-producing animals, rich in antibiotic-resistant bacteria and antibiotic resistance genes (ARGs), poses significant environmental and healthcare risks. Despite global efforts, most manure is not adequately processed before use on fields, escalating the spread of antimicrobial resistance. This study examined how different cattle manure treatments, including composting and storage, affect its microbiome and resistome. The changes occurring in the microbiome and resistome of the treated manure samples were compared with those of raw samples by high-throughput qPCR for ARGs tracking and sequencing of the V3–V4 variable region of the 16S rRNA gene to indicate bacterial community composition. We identified 203 ARGs and mobile genetic elements (MGEs) in raw manure. Post-treatment reduced these to 76 in composted and 51 in stored samples. Notably, beta-lactam, cross-resistance to macrolides, lincosamides and streptogramin B (MLSB), and vancomycin resistance genes decreased, while genes linked to MGEs, integrons, and sulfonamide resistance increased after composting. Overall, total resistance gene abundance significantly dropped with both treatments. During composting, the relative abundance of genes was lower midway than at the end. Moreover, higher biodiversity was observed in samples after composting than storage. Our current research shows that both composting and storage effectively reduce ARGs in cattle manure. However, it is challenging to determine which method is superior, as different groups of resistance genes react differently to each treatment, even though a notable overall reduction in ARGs is observed.

The current study was carried out to detect Salmonella spp. contamination on chicken carcasses and to determine the antibiotic susceptibility profiles and serotype distribution of the isolates. A total of 200 packaged fresh raw chicken samples sold at retail in different markets in central Anatolia were analysed between April 2005 and March 2006. Salmonella spp. was detected in 34% (68/200) of samples using cultural technique and were confirmed by PCR. Ten Salmonella serovars were identified; predominant ones included Typhimurium, Infantis and Heidelberg. All of the Salmonella spp. isolates tested, exhibited resistance to one or more antimicrobial agents used. Resistance to penicillin, oxacillin, clindamycin, vancomycin, erythromycin and ampicillin were evident 100%, 97%, 97%, 92.6%, 89.7% and 85.2%, respectively. Also resistance to tetracycline (67.6%), streptomycin (61.7%), neomycin (55.8%) and cephalothin (52.9%) was observed but a small percentage of the isolates demonstrated resistance to gentamicin (14.7%), chloramphenicol (10.2%), cefotaxime (2.9%) and amikacin (2.9%). As a result, high prevalence of Salmonella spp. and the relatively high resistance among the bacteria tested could pose public health and therapeutic problems in consumers as potential vehicle of resistant Salmonella foodborne infections. To avoid Salmonella contamination, hygienic rules of slaughter and poultry meat processing must be rigorously observed and antibiotic use must be controlled by governmental agencies to prevent increased resistance of antibiotics. © 2010 Elsevier Ltd.

Multidrug-resistant Enterococci colonizing the intestinal tract of patients are the major sources of infection as well as the spread of resistance. The present study aims to determine the antimicrobial resistance profile of Enterococcus species from the intestinal tract of patients admitted to the laboratory of the Yaounde Military Hospital. The study was conducted in patients from July to November 2020. Stool samples were collected and processed for bacterial isolation and antimicrobial susceptibility testing. Stool samples were inoculated onto selective Enterococci media (Bile Esculin azide agar). The isolates obtained were identified to the level of genus and species level: characteristics, Gram staining and use of API 20 Streptococcus system. Susceptibility testing was performed using the Kirby-Bauer solid state disk diffusion method. Of the 35 samples analyzed 19 strains were isolated, 9 Enterococci and 10 non-Enterococci. Enterococci were isolated from 9 (25.7%) of the study subjects. The isolates were Enterococcus faecium (36.9%) followed by Enterococcus faecalis (10.5%), Aerococcus viridans (31.5%), Streptococcus mitis 1 (10.5%) and finally Streptococcus mitis 2 (10.5%). Of the 9 Enterococci isolates tested, 6 (66.6%) were resistant to Ampicillin, 9 (100%) to Gentamycin and 4 (44.4%) to Amikacin. Other alternative antibiotics to treat infection that may be caused by Enterococci have also shown a high rate of resistance in vitro: levofloxacin 6 (66.6%), erythromycin 8 (88.8%), tetracycline 7 (77.7%), chloramphenicol 4 (44.4%). 7 (77.7%) of the Enterococci were resistant to vancomycin. Multidrug resistance was predominant in E. faecium up to R7 (10.5%), however multi resistance was more pronounced at R3 (94.73%). This study reveals a high rate of faecal colonization by multidrug-resistant Enterococci and an increasing prevalence of strains resistant to vancomycin. Thus, periodic monitoring of antibacterial susceptibilities is recommended to detect emerging resistance and prevent the spread of ...

International audience ; The rise of antibiotic resistance is not only a challenge for human and animal health treatments, but is also posing the risk of spreading among bacterial populations in foodstuffs. Farmed fish-related foodstuffs, the food of animal origin most consumed worldwide, are suspected to be a reservoir of antibiotic resistance genes and resistant bacterial hazards. However, scant research has been devoted to the possible sources of diversity in fresh fillet bacterial ecosystems (farm environment including rivers and practices, and factory environment). In this study bacterial communities and the antibiotic resistance genes of fresh rainbow trout fillet were described using amplicon sequencing of the V3-V4 region of the 16S rRNA gene and high-throughput qPCR assay. The antibiotic residues were quantified using liquid chromatography/mass spectrometry methods. A total of 56 fillets (composed of muscle and skin tissue) from fish raised on two farms on the same river were collected and processed under either factory or laboratory sterile filleting conditions. We observed a core-bacterial community profile on the fresh rainbow trout fillets, but the processing conditions of the fillets has a great influence on their mean bacterial load (3.38 ± 1.01 log CFU/g vs 2.29 ± 0.72 log CFU/g) and on the inter-individual diversity of the bacterial community. The bacterial communities were dominated by Gamma- and Alpha-proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria. The most prevalent genera were Pseudomonas, Escherichia-Shigella, Chryseobacterium, and Carnobacterium. Of the 73 antibiotic residues searched, only oxytetracycline residues were detected in 13/56 fillets, all below the European Union maximum residue limit (6.40-40.20 μg/kg). Of the 248 antibiotic resistance genes searched, 11 were found to be present in at least 20% of the fish population (tetracycline resistance genes tetM and tetV, β-lactam resistance genes bla (DHA) and bla (ACC), macrolide resistance gene mphA, vancomycin ...

Legislation aimed at controlling antimicrobial-resistant pathogens through the use of active surveillance cultures to screen hospitalized patients has been introduced in at least 2 US states. In response to the proposed legislation, the Society for Healthcare Epidemiology of America (SHEA) and the Association for Professionals in Infection Control and Epidemiology, Inc., (APIC) have developed this joint position statement. Both organizations are dedicated to combating health care-associated infections with a wide array of methods, including the use of active surveillance cultures in appropriate circumstances. This position statement reviews the proposed legislation and the rationale for use of active surveillance cultures, examines the scientific evidence supporting the use of this strategy, and discusses a number of unresolved issues surrounding legislation mandating use of active surveillance cultures. The following 5 consensus points are offered. (1) Although reducing the burden of antimicrobial-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE), is of preeminent importance, the APIC and the SHEA do not support legislation to mandate use of active surveillance cultures to screen for MRSA, VRE, or other antimicrobial-resistant pathogens. (2) The SHEA and the APIC support the continued development, validation, and application of efficacious and cost-effective strategies for the prevention of infections caused by MRSA, VRE, and other antimicrobial-resistant and antimicrobial-susceptible pathogens. (3) The APIC and the SHEA welcome efforts by health care consumers, together with private, local, state, and federal policy makers, to focus attention on and formulate solutions for the growing problem of antimicrobial resistance and health care-associated infections. (4) The SHEA and the APIC support ongoing additional research to determine and optimize the appropriateness, utility, feasibility, and cost-effectiveness of using active surveillance cultures to screen both lower-risk and high-risk populations. (5) The APIC and the SHEA support stronger collaboration between state and local public health authorities and institutional infection prevention and control experts. © 2007 Association for Professionals in Infection Control and Epidemiology, Inc, and The Society for Healthcare Epidemiology of America. All rights reserved.

Infections in skilled nursing facilities (SNFs) are common and result in frequent hospital transfers, functional decline, and death. Colonization with multidrug-resistant organisms (MDROs) – including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and multidrug-resistant gram-negative bacilli (R-GNB) – is also increasingly prevalent in SNFs. Antimicrobial resistance among common bacteria can adversely affect clinical outcomes and increase health care costs. Recognizing a need for action, legislators, policy-makers, and consumer groups are advocating for surveillance cultures to identify asymptomatic patients with MDROs, particularly MRSA in hospitals and SNFs. Implementing this policy for all SNF residents may be costly, impractical, and ineffective. Such a policy may result in a large increase in the number of SNF residents placed in isolation precautions with the potential for reduced attention by health care workers, isolation, and functional decline. Detection of colonization and subsequent attempts to eradicate selected MDROs can also lead to more strains with drug resistance. We propose an alternative strategy that uses a focused multicomponent bundle approach that targets residents at a higher risk of colonization and infection with MDROs, specifically those who have an indwelling device. If this strategy is effective, similar strategies can be studied and implemented for other high-risk groups.

Multi-drug resistance in Listeria monocytogenes is considered a major public health problem associated with foodborne outbreaks and causes high hospitalization and mortality rates. This study aimed to investigate the antimicrobial resistant genes among Listeria monocytogenes isolated from meat and clinical samples. Phenotypically, the isolates were tested for their susceptibility against the 12 most commonly used antimicrobials in veterinary and human therapy via the disc diffusion method, while conventional PCR was performed to study the presence or absence of 14 resistance genes predicted in L. monocytogenes isolates. The study established that 30(66.66%) of L. monocytogenes isolates showed phenotypic multi-drug resistance against at least three antimicrobial classes. Furthermore, high resistance frequencies were reported among commonly used antibiotics for listeriosis therapy. The present study revealed that the investigated isolates show resistance against tetracycline 33(73.3%), ampicillin 29(64.4%), penicillin 28(62.2%), erythromycin 26(57.8%), and gentamycin, clindamycin and vancomycin 24(53.3% each). Of the 45 L. monocytogenes isolates studied, 37(82.2%) were phenotypically susceptible to meropenem, followed by ciprofloxacin 36(80.0%) and SXT 30(66.7%). PCR amplification of antimicrobial resistance genes established the occurrence of antibiotic resistance genes in all studied L. monocytogenes isolates. Notably, 41(91.11%) of these isolates exhibited more than five resistance genes. Surprisingly, penA and ampC were detected in all L. monocytogenes strains 45(100%), followed by ermB 44(97.8%), tetA 38(84.4%), tetG 32(71.1%), and vanB 30(66.7%). Moreover, vanA 22(48.9%) and tetB 18(40.0%) were detected less frequently. The lowest incidences of resistance genes were observed in L. monocytogenes carrying tetD 4(8.9%) and cmlA 6(13.3%). In conclusion, the study demonstrates that the majority of L. monocytogenes from human and meat samples displayed a high index of resistance to a variety of agents used for clinical listeriosis treatment adding further burden to the existing global antibiotic resistance problem.

The Korean government established an antimicrobial resistance (AMR) surveillance system, compatible with the Global AMR Surveillance System (GLASS): Kor-GLASS. We describe results from the first year of operation of the Kor-GLASS from May 2016 to April 2017, comprising all non-duplicated clinical isolates of major pathogens from blood, urine, faeces and urethral and cervical swabs from six sentinel hospitals. Antimicrobial susceptibility tests were carried out by disk diffusion, Etest, broth microdilution and agar dilution methods. Among 67,803 blood cultures, 3,523 target pathogens were recovered. The predominant bacterial species were Escherichia coli (n = 1,536), Klebsiella pneumoniae (n = 597) and Staphylococcus aureus (n = 584). From 57,477 urine cultures, 6,394 E. coli and 1,097 K. pneumoniae were recovered. Bloodstream infections in inpatients per 10,000 patient-days (10TPD) were highest for cefotaxime-resistant E. coli with 2.1, followed by 1.6 for meticillin-resistant Sta. aureus, 1.1 for imipenem-resistant Acinetobacter baumannii, 0.8 for cefotaxime-resistant K. pneumoniae and 0.4 for vancomycin-resistant Enterococcus faecium. Urinary tract infections in inpatients were 7.7 and 2.1 per 10TPD for cefotaxime-resistant E. coli and K. pneumoniae, respectively. Kor-GLASS generated well-curated surveillance data devoid of collection bias or isolate duplication. A bacterial bank and a database for the collections are under development. ; open

Vancomycin-resistant Enterococcus faecium (VREfm) is a major cause of nosocomial infection and is categorized as high priority by the World Health Organization global priority list of antibiotic-resistant bacteria. In the past, livestock have been proposed as a putative reservoir for drug-resistant E. faecium strains that infect humans, and isolates of the same lineage have been found in both reservoirs. We undertook cross-sectional surveys to isolate E. faecium (including VREfm) from livestock farms, retail meat, and wastewater treatment plants in the United Kingdom. More than 600 isolates from these sources were sequenced, and their relatedness and antibiotic resistance genes were compared with genomes of almost 800 E. faecium isolates from patients with bloodstream infection in the United Kingdom and Ireland. E. faecium was isolated from 28/29 farms; none of these isolates were VREfm, suggesting a decrease in VREfm prevalence since the last UK livestock survey in 2003. However, VREfm was isolated from 1% to 2% of retail meat products and was ubiquitous in wastewater treatment plants. Phylogenetic comparison demonstrated that the majority of human and livestock-related isolates were genetically distinct, although pig isolates from three farms were more genetically related to human isolates from 2001 to 2004 (minimum of 50 single-nucleotide polymorphisms [SNPs]). Analysis of accessory (variable) genes added further evidence for distinct niche adaptation. An analysis of acquired antibiotic resistance genes and their variants revealed limited sharing between humans and livestock. Our findings indicate that the majority of E. faecium strains infecting patients are largely distinct from those from livestock in this setting, with limited sharing of strains and resistance genes.IMPORTANCE The rise in rates of human infection caused by vancomycin-resistant Enterococcus faecium (VREfm) strains between 1988 to the 2000s in Europe was suggested to be associated with acquisition from livestock. As a result, the European Union banned the use of the glycopeptide drug avoparcin as a growth promoter in livestock feed. While some studies reported a decrease in VREfm in livestock, others reported no reduction. Here, we report the first livestock VREfm prevalence survey in the UK since 2003 and the first large-scale study using whole-genome sequencing to investigate the relationship between E. faecium strains in livestock and humans. We found a low prevalence of VREfm in retail meat and limited evidence for recent sharing of strains between livestock and humans with bloodstream infection. There was evidence for limited sharing of genes encoding antibiotic resistance between these reservoirs, a finding which requires further research.

Bodies of water are usually being polluted by wastes from domestic and industrial sources thereby making them unfit for use. Hence, this study aimed at assessing the water quality from Asa River, Ilorin, Nigeria in terms of bacteriological and physicochemical parameters. The bacteriological parameters assessed were heterotrophic bacterial count, total coliform, faecal coliform, identification of the isolates, antibiotic resistance patterns, and plasmid profile of the isolates. Whereas, the assessed physicochemical parameters were pH, total chloride, suspended solid, and total hardness. The heterotrophic bacterial count, total coliform, and faecal coliform counts ranged from 7.6 x 103 to 3.2 x 106 cfu/ml, 9 to 24000 MPN/ 100ml and 0 to 6.0 x 103 cfu/ml respectively. Bacteriological analysis showed that the water contained arrays of bacteria heavily contaminated with pathogens. The genera of bacteria isolated were Escherichia, Pseudomonas, Brevundimonas, Enterococcus, Burkholderia, Enterobacter, Proteus, Pasteurella and Aeromonas. The pH, total hardness, total chloride, and suspended solids of the water ranged from 6.2 to 7.9, 128 to 291 mg/l, 6.21 to 17.04 mg/l and 170 to 310 mg/l respectively. All the Gram-negative bacteria were resistant to cefuroxime, cefixime, augmentin, and ceftazidime. Whereas all the Gram-positive bacteria were resistant to cefuroxime, cloxacillin, augmentin, and vancomycin. The bacteria also showed multiple antibiotic resistance. The resistance of Pasteurella multocida to nitrofurantoin was plasmid mediated. After plasmid curing, Brevundimonas diminuta was susceptible to ciprofloxacin and intermediately susceptible to gentamicin, while Pseudomonas fluorescens was susceptible to ofloxacin and ceftazidime and intermediately susceptible to ciprofloxacin. It was concluded that the water from Asa River at the different sampling points was unsafe in term of bacteriological as well as some physicochemical qualities. It is recommended that this water should not be used in its present state without treatment. Discharge of industrial, agricultural and domestic wastes should be stopped; and open defecation at the bank of this river should be discouraged.

The Korean government established an antimicrobial resistance (AMR) surveillance system, compatible with the Global AMR Surveillance System (GLASS): Kor-GLASS. We describe results from the first year of operation of the Kor-GLASS from May 2016 to April 2017, comprising all non-duplicated clinical isolates of major pathogens from blood, urine, faeces and urethral and cervical swabs from six sentinel hospitals. Antimicrobial susceptibility tests were carried out by disk diffusion, Etest, broth microdilution and agar dilution methods. Among 67,803 blood cultures, 3,523 target pathogens were recovered. The predominant bacterial species were Escherichia coli (n = 1,536), Klebsiella pneumoniae (n = 597) and Staphylococcus aureus (n = 584). From 57,477 urine cultures, 6,394 E. coli and 1,097 K. pneumoniae were recovered. Bloodstream infections in inpatients per 10,000 patient-days (10TPD) were highest for cefotaxime-resistant E. coli with 2.1, followed by 1.6 for meticillin-resistant Sta. aureus, 1.1 for imipenem-resistant Acinetobacter baumannii, 0.8 for cefotaxime-resistant K. pneumoniae and 0.4 for vancomycin-resistant Enterococcus faecium. Urinary tract infections in inpatients were 7.7 and 2.1 per 10TPD for cefotaxime-resistant E. coli and K. pneumoniae, respectively. Kor-GLASS generated well-curated surveillance data devoid of collection bias or isolate duplication. A bacterial bank and a database for the collections are under development.

Die Medikation von Personen nach Exposition oder zur Prophylaxe gegen hochpathogene Bakterien muss schnellstmöglich und effektiv erfolgen, wobei gemäß Empfehlungen Erreger-spezifische Antibiotika verabreicht werden. Bei B. anthracis sind dies Ciprofloxacin, Doxycyclin, Rifampin und Vancomycin; bei Yersinia pestis Ciprofloxacin, Tetracyclin, Streptomycin und Gentamicin. Da die Behandlung der infizierten Personen in der Regel im klinischen Alltag ohne vorherige Überprüfung der Resistenzeigenschaft des Erregers geschieht, wurde eine auf Microarray-Technology basierende Schnelldiagnostik entwickelt, die es ermöglicht innerhalb von 4 - 7 Stunden eine zuverlässige Aussage über vorhandene relevante genetische Resistenzdeterminanten zu treffen. Zu diesem Zweck wurden jeweils für Bacillus anthracis und Yersinia pestis ein Versuchsablauf entwickelt, der zunächst einen DNA-Anreicherungsschritt enthält, der mit einer auf den Erreger abgestimmten Multiplex-PCR realisiert wurde. Im weiteren Verlauf wird eine multiplexe enzymatisch-fluoreszente Markierungsreaktion angeschlossen und das erhaltene Produkt auf den entwickelten Microarrays hybridisiert. Mit diesen ist es nun möglich, sowohl klinisch relevante Resistenzgene nachzuweisen (tet(A-D); tet(K-O); vanA; vanB; aac3-Ia, -IIa, -IVa; ant3-Ia, -IIa; aph6''-Id; aph3''-Ib), als auch Resistent-vermittelnde Mutationen in den Genen gyrA, gyrB, parC bzw. rpoB. Die Evaluation der Microarrays wurde aus politischen und ethischen Gründen nicht mit resistenten B. anthracis oder Y. pestis – Stämmen durchgeführt. Die Microarrays konnten jedoch aufgrund der sehr nahen Verwandtschaft der beiden Organismen zur B. cereus ATCC10987 bzw. Y. pseudotuberculosis DSM8992 mit resistenten Isolaten dieser Spezies evaluiert werden. Dabei wurde sowohl die Detektion der Resistenzgene mit Misch-DNA resistenter Spezies simuliert, als auch die Bestimmung Resistenz-vermittelnden Punktmutationen mit zuvor isolierten und charakterisierten Mutanten geprüft. Im weiteren Verlauf wurden Kinetik und Reproduzierbarkeit der entwickelten Versuchsabläufe untersucht sowie eine Blind-Studie mit klinischen Yersinia enterocolitica Isolaten durchgeführt. ; The medication of persons after exposition or for prophylaxis against highly-pathogenic bacteria must take place as fast as possible and effectively. The following antibiotics are recommended: For B. anthracis are these Ciprofloxacin, Doxycyclin, Rifampin and Vancomycin; for Yersinia pestis Ciprofloxacin, Tetracyclin, Streptomycin and Gentamicin. Treatment of infected persons is usually realized in clinical diagnostic without previous examination of resistance of the infecting species. In front of this background an assay was developed based on microarray-technology, which enables the obtaining of reliable results of existing relevant genetic resistance determinants within 4 - 7 hours. Two separate assays for Bacillus anthracis and Yersinia pestis were developed, which harbour a DNA enrichment-step, -realized by a multiplex PCR compatible with the examined species-, subsequently a multiplex enzymatic fluorescent labelling and hybridization using the developed diagnostic microarrays. Using these microarrays it is possible to detect clinically relevant resistance genes (tet (A-D); tet (K-O); vanA; vanB; aac3-Ia, - IIa, - IVa; ant3-Ia, - IIa; aph6'' - Id; aph3'' - Ib) and resistance-mediating point-mutations in genes gyrA, gyrB, parC and/or rpoB in one single step. Because of political and ethical reasons evaluation of the microarrays was not performed using resistant B. anthracis- or Y. pestis - isolates. The microarrays could be evaluated because of a strong relationship of these two organisms to the non-highly-pathogenic organisms B. cereus ATCC10987 and Y. pseudotuberculosis DSM8992. The detection of both, resistance genes and resistance-obtaining point mutations, was performed using in this study isolated and well characterized mutants or mixture-DNA of resistant species. In further process kinetics and reproducibility of the developed assay were investigated as well as a blind panel was performed using clinical Yersinia enterocolitica-isolates.

Introduction Antimicrobial resistance (AMR) has a critical global impact, mostly affecting low- and middle-income countries. A major knowledge gap exists in understanding the transmission pathway of the gut colonisation with AMR bacteria between healthy humans and their animals in addition to the presence of those AMR bacteria in the surrounding environment. A One Health (OH) approach is necessary to address this multisectoral problem. Methods and analysis This cross-sectional, mixed-method OH study design will use both quantitative and qualitative methods of data collection. Quantitative methods will be carried out to assess the prevalence and risk factors associated with multidrug resistant Gram-negative bacteria and vancomycin-resistant enterococci in humans, animals (cattle) and the environment. The focus will be on cattle rearing as an exposure risk for AMR among humans. The assessment of AMR in the population of Jimma, Ethiopia with or without exposure to cattle will reinforce the importance of OH research to identify the impending exchange of resistance profile between humans and animals as well as its ultimate dissemination in the surrounding environment. The targeted semistructured key stakeholder interviews will aid to strengthen the OH-AMR surveillance in Ethiopia by understanding the acceptability of an integrated AMR surveillance platform based on the District Health Information Software-2 and the feasibility of its context-specific establishment. Ethics and dissemination The study has been approved by the Regional Ethics Committee, Norway, and the Institutional Review Board of Jimma University, Ethiopia. The study's data will be stored on a secure server known as Services for Sensitive Data hosted by the University of Oslo. In addition, the new European Union Global Data Protection Guidelines for data sharing, storage and protection will be followed. We will publish the results in peer-reviewed journals and present the findings at national and international conferences. ; Understanding transmission ...