ESKAPE Bacteria and Extended-Spectrum-β-Lactamase-Producing Escherichia coli Isolated from Wastewater and Process Water from German Poultry Slaughterhouses

Abstract

The wastewater of livestock slaughterhouses is considered a source of antimicrobial-resistant bacteria with clinical relevance and may thus be important for their dissemination into the environment. To get an overview of their occurrence and characteristics, we investigated process water (n = 50) from delivery and unclean areas as well as wastewater (n = 32) from the in-house wastewater treatment plants (WWTPs) of two German poultry slaughterhouses (slaughterhouses S1 and S2). The samples were screened for ESKAPE bacteria (Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) and Escherichia coli. Their antimicrobial resistance phenotypes and the presence of extended-spectrum-β-lactamase (ESBL), carbapenemase, and mobilizable colistin resistance genes were determined. Selected ESKAPE bacteria were epidemiologically classified using different molecular typing techniques. At least one of the target species was detected in 87.5% (n = 28/32) of the wastewater samples and 86.0% (n = 43/50) of the process water samples. The vast majority of the recovered isolates (94.9%, n = 448/472) was represented by E. coli (39.4%), the A. calcoaceticus-A. baumannii (ACB) complex (32.4%), S. aureus (12.3%), and K. pneumoniae (10.8%), which were widely distributed in the delivery and unclean areas of the individual slaughterhouses, including their wastewater effluents. Enterobacter spp., Enterococcus spp., and P. aeruginosa were less abundant and made up 5.1% of the isolates. Phenotypic and genotypic analyses revealed that the recovered isolates exhibited diverse resistance phenotypes and β-lactamase genes. In conclusion, wastewater effluents from the investigated poultry slaughterhouses exhibited clinically relevant bacteria (E. coli, methicillin-resistant S. aureus, K. pneumoniae, and species of the ACB and Enterobacter cloacae complexes) that contribute to the dissemination of clinically relevant resistances (i.e., blaCTX-M or blaSHV and mcr-1) in the environment. IMPORTANCE Bacteria from livestock may be opportunistic pathogens and carriers of clinically relevant resistance genes, as many antimicrobials are used in both veterinary and human medicine. They may be released into the environment from wastewater treatment plants (WWTPs), which are influenced by wastewater from slaughterhouses, thereby endangering public health. Moreover, process water that accumulates during the slaughtering of poultry is an important reservoir for livestock-associated multidrug-resistant bacteria and may serve as a vector of transmission to occupationally exposed slaughterhouse employees. Mitigation solutions aimed at the reduction of the bacterial discharge into the production water circuit as well as interventions against their further transmission and dissemination need to be elaborated. Furthermore, the efficacy of in-house WWTPs needs to be questioned. Reliable data on the occurrence and diversity of clinically relevant bacteria within the slaughtering production chain and in the WWTP effluents in Germany will help to assess their impact on public and environmental health.

The Animal-foods-environment interface of Klebsiella pneumoniae in Germany: an observational study on pathogenicity, resistance development and the current situation

“… K. pneumonia in environmental sources (10 studies) Wastewater from poultry slaughterhouses 51 ESBL-producing K. pneumoniae and Six colistin resistance Strains harbored bla SHV-1, 2, 25, 27, and 28, combinations SHV-2-TEM-1b and SHV-27-TEM-52b and CTX-M-1 were detected. Three strains with mcr -1 were isolated ND December 2016–September 2018 Savin et al 2020 [ 66 ] Water/sediment samples 9 MDR-ESBL producing strains Strains harbored bla SHV–28, bla CTX–M–15, tet (D), cat B3, aac (3)-IId, str A, str B, fos A, ere (A), sul 1, sul 2, oqx A, oqx B, aac (6′)Ib-cr, qnr S1, dfr A5 and dfr A14 Northern Germany 2017 Falgenhauer et al 2019 [ 65 ] Raw sewage one ESBL producing strain 182 (WGS of virulent phage PMBT1). Resistant patterns not determined ND Kiel ND Koberg et al 2017 [ 64 ] Washing machines, dishwashers ND β-Lactamase genes AmpC- and OXA-type were detected North-Rhine-Westphalia ND Rehberg et al 2017 [ 63 ] Wastewater treatment plants ND Colistin resistance, tetracycline resistance and erythromycin resistance Strains harbored mcr -1, tet M, erm B gene, ß-lactam ( bla CTX-M-32, ...…”

An Investigation of Antibacterial Resistance Patterns in Isolated Bacteria from Contaminated Water Samples in Poultry Slaughterhouses

“…The results showed that among these isolates, also were observed cases with multiple resistances. Savin et al (2020) reported that wastewater effluents from the poultry slaughterhouses exhibited clinically relevant bacteria (E. coli (39.4%), methicillin-resistant S.aureus (12.3%), K. pneumonia (10.8%), and species of the ACB (32.4%), which were widely distributed in their wastewater effluents (Savin et al, 2020).…”

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

“…The wastewater from slaughterhouses is considered a relevant source of antimicrobial resistant bacteria and consequently may be important for its diffusion into the environment [ 46 ]. The livestock pressure in most of the Spanish rivers studied in this work was high, where pig farms stood out mainly ( Table S1 ).…”

“…In addition, the association between bla OXA-1 and resistance to aminoglycosides and quinolones reported by Osińska et al, 2016 [ 48 ] was confirmed in the present study, because the two WWTP isolates carrying bla OXA-1 were resistant to TO, LV, CIP, MXF and NOR. Indeed, the increased quinolone resistance rate in the isolates (50%) may be caused by the use of enrofloxacin in slaughtered broiler herds [ 46 ]. Furthermore, the high prevalence regarding AmpC β-lactamases could be related with the large number of Serratia and Citrobacter strains (carriers of chromosomal AmpC).…”