Antibiotic-Resistant Enterobacteriaceae in Wastewater of Abattoirs

Antibiotic-resistant Enterobacteriaceae are regularly detected in livestock. As pathogens, they cause difficult-to-treat infections and, as commensals, they may serve as a source of resistance genes for other bacteria. Slaughterhouses produce significant amounts of wastewater containing antimicrobial-resistant bacteria (AMRB), which are released into the environment. We analyzed the wastewater from seven slaughterhouses (pig and poultry) for extended-spectrum β-lactamase (ESBL)-carrying and colistin-resistant Enterobacteriaceae. AMRB were regularly detected in pig and poultry slaughterhouse wastewaters monitored here. All 25 ESBL-producing bacterial strains (19 E. coli and six K. pneumoniae) isolated from poultry slaughterhouses were multidrug-resistant. In pig slaughterhouses 64% (12 of 21 E. coli [57%] and all four detected K. pneumoniae [100%]) were multidrug-resistant. Regarding colistin, resistant Enterobacteriaceae were detected in 54% of poultry and 21% of pig water samples. Carbapenem resistance was not detected. Resistant bacteria were found directly during discharge of wastewaters from abattoirs into water bodies highlighting the role of slaughterhouses for environmental surface water contamination.

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Detection of Antibiotic-Resistant Bacteria, Resistance Determinants, and Mobile Elements in Surface Waters in Lebanon

10.1101/2021.02.12.21251645 ◽

2021 ◽

Author(s):

Jennifer Moussa ◽

Edmond Abboud ◽

Sima Tokajian

Keyword(s):

Surface Water ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Population Mobility ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Resistance Determinants ◽

Bacterial Contaminants ◽

Plasmid Analysis

AbstractThe prevalence of antibiotic-resistant bacteria in surface water in Lebanon is a growing concern and understanding the mechanisms of the spread of resistance determinants is essential. We aimed at studying the occurrence of resistant organisms and determinants in surface water sources in Lebanon and understanding their mobilization and transmission. Water samples were collected from five major rivers in Lebanon. 91 isolates were recovered out of which 25 were multidrug-resistant (MDR) and accordingly were further characterized. Escherichia coli and Klebsiella pneumoniae were the most commonly identified MDR isolates. Conjugation assays coupled with in silico plasmid analysis were performed and validated using PCR-based replicon typing (PBRT) to identify and confirm incompatibility groups and the localization of β-lactamase encoding genes. E. coli EC23 carried a blaNDM-5 gene on a conjugative, multireplicon plasmid, while blaCTX-M-15 and blaTEM-1B were detected in the majority of the MDR isolates. Different ST types were identified including the highly virulent E. coli ST131. Our results showed a common occurrence of bacterial contaminants in surface water and an increase in the risk for the dissemination of resistance determinants exacerbated with the ongoing intensified population mobility in Lebanon and the widespread lack of wastewater treatment.

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Tryptophan Silver Nanoparticles Synthesized by Photoreduction Method: Characterization and Determination of Bactericidal and Anti-Biofilm Activities on Resistant and Susceptible Bacteria

International Journal of Tryptophan Research ◽

10.1177/1178646919831677 ◽

2019 ◽

Vol 12 ◽

pp. 117864691983167 ◽

Cited By ~ 7

Author(s):

Daniella dos Santos Courrol ◽

Carla Regina Borges Lopes ◽

Camila Bueno Pacheco Pereira ◽

Marcia Regina Franzolin ◽

Flávia Rodrigues de Oliveira Silva ◽

...

Keyword(s):

Silver Nanoparticles ◽

Biofilm Formation ◽

Antimicrobial Agents ◽

Irradiation Time ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

E Coli ◽

Static Conditions

The high rates of antibiotics use in hospitals have resulted in a condition where multidrug-resistant pathogens have become a severe threat to the human health worldwide. Therefore, there is an increasing necessity to identify new antimicrobial agents that can inhibit the multidrug-resistant bacteria and biofilm formation. In this study, antibacterial and anti-biofilm activities of tryptophan silver nanoparticles (TrpAgNP) were investigated. The TrpAgNPs were synthesized by photoreduction method, and the influence of irradiation time and concentration of reagents were analyzed. The nanoparticles were characterized by transmission electron microscopy, Zeta Potential and (UV)-absorption spectra. The antibacterial activity of TrpAgNPs was tested for antibiotic-resistant and susceptible pathogens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Salmonella typhimurium, and Pseudomonas aeruginosa, evaluating the influence of photoreduction parameters in bactericidal effect. The results have shown that TrpAgNPs solutions with lower tryptophan/silver nitrate (AgNO3) ratio and higher AgNO3 concentration have higher bactericidal action against bacteria with inhibition of ~100% in almost all studied bacterial strains. The antimicrobial activity of TrpAgNPs within biofilms generated under static conditions of antibiotic-resistant and susceptible strains of S. aureus, S. epidermidis, E. coli, K. pneumoniae, C. freundii, and P. aeruginosa was also investigated. The results showed that TrpAgNPs have an inhibitory effect against biofilm formation, exceeding 50% in the case of Gram-negative bacteria ( E. coli, K. pneumoniae, C. freundii, and P. aeruginosa—54.8% to 98.8%). For Gram-positive species, an inhibition of biofilm formation of 68.7% to 72.2 % was observed for S. aureus and 20.0% to 40.2% for S. epidermidis.

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Multiresistant Bacteria Isolated from Intestinal Faeces of Farm Animals in Austria

Antibiotics ◽

10.3390/antibiotics10040466 ◽

2021 ◽

Vol 10 (4) ◽

pp. 466

Author(s):

Herbert Galler ◽

Josefa Luxner ◽

Christian Petternel ◽

Franz F. Reinthaler ◽

Juliana Habib ◽

...

Keyword(s):

Meat Products ◽

Animal Husbandry ◽

Multidrug Resistant ◽

Farm Animals ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Multiresistant Bacteria ◽

Vancomycin Resistant

In recent years, antibiotic-resistant bacteria with an impact on human health, such as extended spectrum β-lactamase (ESBL)-containing Enterobacteriaceae, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE), have become more common in food. This is due to the use of antibiotics in animal husbandry, which leads to the promotion of antibiotic resistance and thus also makes food a source of such resistant bacteria. Most studies dealing with this issue usually focus on the animals or processed food products to examine the antibiotic resistant bacteria. This study investigated the intestine as another main habitat besides the skin for multiresistant bacteria. For this purpose, faeces samples were taken directly from the intestines of swine (n = 71) and broiler (n = 100) during the slaughter process and analysed. All samples were from animals fed in Austria and slaughtered in Austrian slaughterhouses for food production. The samples were examined for the presence of ESBL-producing Enterobacteriaceae, MRSA, MRCoNS and VRE. The resistance genes of the isolated bacteria were detected and sequenced by PCR. Phenotypic ESBL-producing Escherichia coli could be isolated in 10% of broiler casings (10 out of 100) and 43.6% of swine casings (31 out of 71). In line with previous studies, the results of this study showed that CTX-M-1 was the dominant ESBL produced by E. coli from swine (n = 25, 83.3%) and SHV-12 from broilers (n = 13, 81.3%). Overall, the frequency of positive samples with multidrug-resistant bacteria was lower than in most comparable studies focusing on meat products.

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Retail Chicken Carcasses as a Reservoir of Antimicrobial- Resistant Escherichia coli

University of the Future: Re-Imagining Research and Higher Education ◽

10.29117/quarfe.2020.0115 ◽

2020 ◽

Author(s):

Nahla Omer Eltai ◽

Hadi M. Yassine ◽

Sara H. Al-Hadidi ◽

Tahra ElObied ◽

Asmaa A. Al Thani ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Growth Promotion ◽

Multidrug Resistant ◽

Chicken Meat ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Inappropriate Use ◽

Esbl Producers

The dissemination of antimicrobial resistance (AMR) bacteria has been associated with the inappropriate use of antibiotics in both humans and animals and with the consumption of food contaminated with resistant bacteria. In particular, the use of antibiotics as prophylactic and growth promotion purposes in food-producing animals has rendered many of the antibiotics ineffective. The increased global prevalence of AMR poses a significant threat to the safety of the world’s food supply. Objectives: This study aims at determining the prevalence of antibiotic-resistant Escherichia coli (E. coli) isolated from local and imported retail chicken meat in Qatar. Methodology: A total of 270 whole chicken carcasses were obtained from three different hypermarket stores in Qatar. A total of 216 E. coli were isolated and subjected to antibiotic susceptibility testing against 18 relevant antibiotics using disc diffusion and micro- dilution methods. Furthermore, extended-spectrum β-lactamase (ESBL) production was determined via a double-disc synergetic test. Isolates harboring colistin resistance were confirmed using multiplex-PCR and DNA sequencing. Results: Nearly 89% (192/216) of the isolates were resistant to at least one antibiotics. In general, isolates showed relatively higher resistance to sulfamethoxazole (62%), tetracycline (59.7%), ampicillin and trimethoprim (52.3%), ciprofloxacin (47.7%), cephalothin, and colistin (31.9%). On the other hand, less resistance was recorded against amoxicillin/clavulanic acid (6%), ceftriaxone (5.1%), nitrofurantoin (4.2%) and piperacillin/tazobactam (4.2%), cefepime (2.3%), meropenem (1.4%), ertapenem (0.9%), and amikacin (0.9%). Nine isolates (4.2%) were ESBL producers. Furthermore, 63.4% were multidrug-resistant (MDR). The percentage of MDR, ESBL producers, and colistin-resistant isolates was significantly higher among local isolates compared to imported chicken samples. Conclusion: We reported a remarkably high percentage of the antibiotic-resistant E. coli in chicken meat sold at retail in Qatar. The high percentage of MDR and colistin isolates is troublesome to the food safety of raw chicken meat and the potential of antibiotic resistance spread to public health. Our findings support the need for the implementation of one health approach to address the spread of antimicrobial resistance and the need for a collaborative solution.

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Prevalence of Multidrug-Resistant Foodborne Pathogens and Indicator Bacteria from Edible Offal and Muscle Meats in Nashville, Tennessee

Foods ◽

10.3390/foods9091190 ◽

2020 ◽

Vol 9 (9) ◽

pp. 1190

Author(s):

Siqin Liu ◽

Agnes Kilonzo-Nthenge ◽

Samuel N. Nahashon ◽

Bharat Pokharel ◽

Abdullah Ibn Mafiz ◽

...

Keyword(s):

Foodborne Pathogens ◽

Salmonella Enterica ◽

Ground Beef ◽

Multidrug Resistant ◽

Indicator Bacteria ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Beef Liver ◽

E Coli ◽

Retail Meats

This study investigated the prevalence of antimicrobial-resistant bacteria in retail edible offal and muscle meats in Nashville, Tennessee. A total of 348 retail meats (160 edible offal and 188 muscle) were analyzed for Salmonella enterica serovar, Campylobacter, Escherichia coli, E. coli O157:H7, and enterococci. Bacteria was identified using biochemical and PCR methods. Salmonella enterica serovar (4.4% and 4.3%), Campylobacter (1.9% and 1.1%), E. coli (79.4% and 89.4%), and enterococci (88.1% and 95.7%) was detected in offal and muscle meats, respectively. Chicken liver (9.7%) was most frequently contaminated with Salmonella enterica serovar, followed by ground chicken (6.9%) and chicken wings (4.2%). No Salmonella enterica serovar was detected in beef liver, beef tripe, and ground beef. The prevalence of Campylobacter was 6.9%, 2.3%, and 1.4% in beef liver, ground beef, and ground chicken, respectively. None of the meats were positive for E. coli O157:H7. Resistance of isolates was significantly (p < 0.05) highest in erythromycin (98.3%; 99.1%), followed by tetracycline (94%; 98.3%), vancomycin (88.8%; 92.2%) as compared to chloramphenicol (43.1%; 53.9%), amoxicillin/clavulanic (43.5%; 45.7%), and ciprofloxacin (45.7%; 55.7%) in offal and muscle meats, respectively. Imipenem showed the lowest resistance (0%; 0.9%). A total of 41 multidrug-resistant patterns were displayed. Edible offal could be a source of antibiotic-resistant bacteria.

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Microbial Isolates from Vegetable Foreign Bodies Inhaled by Dogs

Veterinary Medicine International ◽

10.1155/2018/3089282 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 2

Author(s):

Sara Flisi ◽

Manuel Dall’Aglio ◽

Costanza Spadini ◽

Clotilde Silvia Cabassi ◽

Fausto Quintavalla

Keyword(s):

Respiratory Tract ◽

Antibiotic Susceptibility ◽

Resistant Bacteria ◽

Upper Respiratory Tract ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

E Coli ◽

Bacteria And Fungi ◽

Susceptibility Tests ◽

The Right

Grass-seed inhalation is a common problem in canine patients, in particular during summer months, migrating in upper and lower respiratory tract. Grass awns can harbor bacteria and fungi, causing grass seeds foreign body-related disease (GSFBD). Aim of this study was to investigate the aerobic microbial flora isolated from grass awns extracted from 41 dogs with GSFBD and the antibiotic susceptibility of the isolated bacterial strains. Fifty-four grass awns were localized with diagnostic imaging tests and removed by endoscopy from respiratory tract. The most frequent localizations were in the left nostril and the right hemithorax. Only one grass awn was extracted from each patient except in 7 that had more than one. Bacteriological and mycological cultures, strains identification, and antibiotic susceptibility tests were performed. One or more bacterial strains were isolated from all grass awns. Fungal strains were isolated only in 4 cases. Staphylococcussp. was the most frequent isolate in the upper respiratory tract (36.8%), whileE. coli(24.4%) was the most frequent isolate in the lower tract. Fluoroquinolones and Doxycycline were the most effective antibiotics, while resistance was observed against Gentamicin (>93%), Cefapirin, and Clindamycin (>80%). These data are relevant in relation to the use of these antibiotics in both animals and humans, for the risk of transmission of antibiotic resistant bacteria or resistance genes.

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Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against Staphylococcus aureus

Processes ◽

10.3390/pr8121587 ◽

2020 ◽

Vol 8 (12) ◽

pp. 1587

Author(s):

Feng Wang ◽

Xiaohang Liu ◽

Zhengyu Deng ◽

Yao Zhang ◽

Xinyu Ji ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Bacterial Infections ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Conventional Antibiotics ◽

Phage Lysin ◽

Shed Light

With the increasing spread of multidrug-resistant bacterial pathogens, it is of great importance to develop alternatives to conventional antibiotics. Here, we report the generation of a chimeric phage lysin, MLTphg, which was assembled by joining the lysins derived from Meiothermus bacteriophage MMP7 and Thermus bacteriophage TSP4 with a flexible linker via chimeolysin engineering. As a potential antimicrobial agent, MLTphg can be obtained by overproduction in Escherichia coli BL21(DE3) cells and the following Ni-affinity chromatography. Finally, we recovered about 40 ± 1.9 mg of MLTphg from 1 L of the host E. coli BL21(DE3) culture. The purified MLTphg showed peak activity against Staphylococcus aureus ATCC6538 between 35 and 40 °C, and maintained approximately 44.5 ± 2.1% activity at room temperature (25 °C). Moreover, as a produced chimera, it exhibited considerably improved bactericidal activity against Staphylococcus aureus (2.9 ± 0.1 log10 reduction was observed upon 40 nM MLTphg treatment at 37 °C for 30 min) and also a group of antibiotic-resistant bacteria compared to its parental lysins, TSPphg and MMPphg. In the current age of growing antibiotic resistance, our results provide an engineering basis for developing phage lysins as novel antimicrobial agents and shed light on bacteriophage-based strategies to tackle bacterial infections.

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Development of Staphylococcus Enzybiotics: The Ph28 Gene of Staphylococcus epidermidis Phage PH15 Is a Two-Domain Endolysin

Antibiotics ◽

10.3390/antibiotics9040148 ◽

2020 ◽

Vol 9 (4) ◽

pp. 148

Author(s):

Magdy Mohamed Muharram ◽

Ashraf Tawfik Abulhamd ◽

Mohammed F. Aldawsari ◽

Mohamed Hamed Alqarni ◽

Nikolaos E. Labrou

Keyword(s):

Staphylococcus Epidermidis ◽

Multidrug Resistant ◽

Health Concern ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Evolutionary Features ◽

Ncbi Accession ◽

Molecular Modelling Studies ◽

Modelling Studies

Given the worldwide increase in antibiotic resistant bacteria, bacteriophage derived endolysins represent a very promising new alternative class of antibacterials in the fight against infectious diseases. Endolysins are able to degrade the prokaryotic cell wall, and therefore have potential to be exploited for biotechnological and medical purposes. Staphylococcus epidermidis is a Gram-positive multidrug-resistant (MDR) bacterium of human skin. It is a health concern as it is involved in nosocomial infections. Genome-based screening approach of the complete genome of Staphylococcus virus PH15 allowed the identification of an endolysin gene (Ph28; NCBI accession number: YP_950690). Bioinformatics analysis of the Ph28 protein predicted that it is a two-domain enzyme composed by a CHAP (22-112) and MurNAc-LAA (171-349) domain. Phylogenetic analysis and molecular modelling studies revealed the structural and evolutionary features of both domains. The MurNAc-LAA domain was cloned, and expressed in E. coli BL21 (DE3). In turbidity reduction assays, the recombinant enzyme can lyse more efficiently untreated S. epidermidis cells, compared to other Staphylococcus strains, suggesting enhanced specificity for S. epidermidis. These results suggest that the MurNAc-LAA domain from Ph28 endolysin may represent a promising new enzybiotic.

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Plasmid-mediated ciprofloxacin resistance imparts a selective advantage on Escherichia coli ST131

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02146-21 ◽

2021 ◽

Author(s):

Minh-Duy Phan ◽

Kate M. Peters ◽

Laura Alvarez Fraga ◽

Steven C. Wallis ◽

Steven Hancock ◽

...

Keyword(s):

Escherichia Coli ◽

Carbapenem Resistance ◽

Bloodstream Infections ◽

Selective Advantage ◽

Multidrug Resistant ◽

Resistance Mutations ◽

Gene Encoding ◽

Antibiotic Resistant ◽

E Coli ◽

Rapid Dissemination

Escherichia coli ST131 is a recently emerged antibiotic resistant clone responsible for high rates of urinary tract and bloodstream infections. Despite its global dominance, the precise mechanisms that have driven the rapid dissemination of ST131 remain unknown. Here, we show that the plasmid-associated resistance gene encoding the AAC(6’)-Ib-cr enzyme that inactivates the fluoroquinolone antibiotic ciprofloxacin is present in >70% of strains from the most rapidly expanding subgroup of multidrug resistant ST131. Using a series of genome-edited and plasmid-cured isogenic strains, we demonstrate that the aac(6’)-Ib-cr gene confers a selective advantage on ST131 in the presence of ciprofloxacin, even in strains containing chromosomal GyrA and ParC FQ-resistance mutations. Further, we identify a pattern of emerging carbapenem resistance in other common E. coli clones carrying both aac(6’)-Ib-cr and chromosomal FQ-resistance mutations, suggesting this dual resistance combination may also impart a selective advantage on these non-ST131 antibiotic resistant lineages.

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1004. Cladophora in Lake Michigan May Serve as Important Reservoirs for Antibiotic-Resistant Bacteria

Open Forum Infectious Diseases ◽

10.1093/ofid/ofab466.1198 ◽

2021 ◽

Vol 8 (Supplement_1) ◽

pp. S592-S593

Author(s):

Dannielle C Grayer ◽

Latania K Logan

Keyword(s):

Surface Water ◽

Water Samples ◽

Lake Michigan ◽

Primary Objective ◽

Resistant Bacteria ◽

Research Station ◽

Antibiotic Resistant ◽

E Coli ◽

Amoxicillin Clavulanate ◽

Surface Water Samples

Abstract Background Cladophora is a green algae, native to the Great Lakes, and found in large quantities along Lake Michigan shorelines. Previous studies have shown that Cladophora provide protection and nutrients for the Enterobacteriaceae (Ent) family, allowing persistence and regrowth. Chicago waterways harbor concerning antibiotic-resistant (AR) Ent, however the community reservoirs are unknown. Our primary objective was to assess whether Cladophora harbor AR Ent and to secondarily assess AR Ent in local beach waters where Cladophora are present. Figure 1. Map of Lake Michigan showing sites (S1-S3) where Cladophora samples were collected. NB, North Beach, Racine, Wisconsin; Michigan City, Indiana; PL, Portage Lakefront, Indiana Dunes National Park, Indiana. Figure 2. Map of Lake Michigan showing sites (S4-S7) where beach surface water samples were obtained. Montrose, Foster, 63rd St, & Calumet beaches in Chicago, Illinois. Methods Cladophora were processed from three Indiana Lake Michigan sites (S1 and S2; Fig 1) in 2002 and 2012 (S3; Fig 1) at the USGS Lake Michigan Ecological Research Station (Chesterton, IN). In 2015, surface water samples were obtained by the USGS at four Chicago beaches (S4-S7; Fig 2), which also amass Cladophora. Bacteria were isolated shortly after collection. In 2019-2020, Ent were cultivated and susceptibilities were performed at Rush. Results In 2002-2003 (S1 and S2), 160 E. coli were cultured from Cladophora. There was AR to multiple classes, highest overall in tetracyclines (7.5%, range 6.2%-18.7%), cefoxitin (8%), and cefazolin (5.6%). Resistance to cefuroxime was 0.6%. Four Salmonella isolates from 2012 (S3) were pan-susceptible, while two Citrobacter isolates were resistant to penicillins, 1st and 2nd generation cephalosporins, and cephamycins. Beach surface water samples from 2015 revealed more pronounced AR in E. coli (n=185) involving multiple classes, including highest in ampicillin (12.4%), tetracyclines (8.1%); piperacillin (7%); cefazolin (3.8%), cephamycins (3.2%) and amoxicillin-clavulanate (2.7%). Resistance to 3rd-generation cephalosporins, fluoroquinolones, trimethoprim/sulfamethoxazole ranged from 0.5-2%. AR Ent varied by beach site with highest percentages at S4, the only site with an associated dog beach. Conclusion These findings suggest that Cladophora in recreational waterways may serve as reservoirs for AR Ent. Differences in AR Ent at beach sites may reflect varying degrees of fecal contamination. Identifying community reservoirs is key to better understanding the acquisition of antibiotic resistant Ent among healthy populations and has long-term ecological and public health implications. Disclosures All Authors: No reported disclosures

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