scholarly journals Molecular Analysis of Antimicrobial Resistance among Enterobacteriaceae Isolated from Diarrhoeic Calves in Egypt

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1712
Author(s):  
Abdel-Moamen E. Meshref ◽  
Ibrahim E. Eldesoukey ◽  
Abdulaziz S. Alouffi ◽  
Saleh A. Alrashedi ◽  
Salama A. Osman ◽  
...  

The present study was designed to investigate the presence of genes that conferred resistance to antimicrobials among Enterobacteriaceae that were isolated from diarrhoeic calves. A total of 120 faecal samples were collected from diarrhoeic calves that were raised in Kafr El-Sheikh governorate, Egypt. The samples were screened for Enterobacteriaceae. A total of 149 isolates of bacteria were recovered and identified; Escherichia coli was found to be the most overwhelming species, followed by Citrobacter diversus, Shigella spp., Serratia spp., Providencia spp., Enterobacter spp., Klebsiella pneumoniae, Proteus spp., Klebsiella oxytoca, and Morganella morganii. All isolates were tested for susceptibility to 12 antimicrobials; resistant and intermediately resistant strains were screened by conventional polymerase chain reaction for the presence of antimicrobial resistance genes. Of the 149 isolates, 37 (24.8%) exhibited multidrug resistant phenotypes. The most prevalent multidrug resistant species were E. coli, C. diversus, Serratia spp., K. pneumoniae, Shigella spp., Providencia spp., and K. oxytoca. Class 1 integrons were detected in 28 (18.8%) isolates. All isolates were negative for class 2 integrons. The blaTEM gene was identified in 37 (24.8%) isolates, whereas no isolates carried the blaCTX-M gene. The florfenicol gene (floR) was detected in two bacterial isolates (1.3%). The findings of this study reveal that calves may act as potential reservoirs of multidrug resistant bacteria that can be easily transmitted to humans.

2017 ◽  
Vol 80 (12) ◽  
pp. 2048-2055 ◽  
Author(s):  
Tao Yu ◽  
Xiaobing Jiang ◽  
Yu Liang ◽  
Yanping Zhu ◽  
Jinhe Tian ◽  
...  

ABSTRACT The aim of this study was to investigate antimicrobial resistance and the presence and transferability of corresponding resistance genes and integrons in bacteria isolated from cooked meat samples in the People's Republic of China. A total of 150 isolates (22 species belonging to 15 genera) were isolated from 49 samples. Resistance of these isolates to antimicrobials was commonly observed; 42.7, 36.0, and 25.3% of the isolates were resistant to tetracycline, streptomycin, and ampicillin, respectively. Multidrug resistance was observed in 41 (27.3%) of the isolates. Sixteen resistance genes, i.e., blaTEM-1 and blaCTX-M-14 (β-lactams), aac(3)-IIa (gentamicin), strA and strB (streptomycin), qnrB and qnrS (fluoroquinolone), sul1, sul2, and sul3 (sulfamethoxazole), cat1 and cat2 (chloramphenicol), and tetM, tetA, tetS, and tetB (tetracycline), were found in 54 isolates. One isolate of Pseudomonas putida carried qnrB, and sequence analysis of the PCR product revealed 96% identity to qnrB2. The qnr genes were found coresiding and were cotransferred with bla genes in two isolates. Twelve isolates were positive for the class 1 integrase gene, and four isolates carried the class 2 integrase gene. However, no class 3 integrase gene was detected. One isolate of Proteus mirabilis carried dfrA32-ereA-aadA2, and this unusual array could be transferred to Escherichia coli. Nonclassic class 1 integrons lacking qacEΔ1 and sul1 genes were found in 2 of the 12 intI1-positive isolates. Our results revealed the presence of multidrug-resistant bacteria in cooked meats and the presence and transferability of resistance genes in some isolates, suggesting that cooked meat products may act as reservoirs of drug-resistant bacteria and may facilitate the spread of resistance genes.


2019 ◽  
Vol 82 (7) ◽  
pp. 1183-1190
Author(s):  
SE HYUN SON ◽  
KWANG WON SEO ◽  
YEONG BIN KIM ◽  
HYE YOUNG JEON ◽  
EUN BI NOH ◽  
...  

ABSTRACT Edible offal, which is the nonmuscular part of the livestock, is a popular food product in many countries. However, it can be easily contaminated by bacteria, such as Escherichia coli, during slaughter and processing and regarded as a reservoir for transfer of antimicrobial-resistant bacteria to humans. This study aimed to investigate prevalence and characteristics of antimicrobial-resistant E. coli isolates from edible offal in Korea. A total of 320 chicken offal samples, 540 pig offal samples, and 560 cattle offal samples were collected. Among the 118 E. coli isolates obtained, resistance to at least one antimicrobial agent was revealed in 32 (100%), 46 (95.8%), and 26 (68.4%) isolates of chicken, pig, and cattle offals, respectively, with an overall prevalence of 88.1% (104 of 118). The isolates from chicken offal showed highest resistance to most antimicrobial agents, with the exception of higher ampicillin resistance for isolates from pig offal. In the distribution of antimicrobial resistance genes of 69 (58.5%) multidrug-resistant (MDR) E. coli, blaTEM-1 (97.1%), tetA (76.6%), sul2 (70.6%), and cmlA (57.4%) were most prevalent. Class 1 and class 2 integrons were detected in 82.6 and 2.9% of the MDR isolates, respectively. In total, seven virulence genes (eaeA, escV, astA, fimH, papC, sfa/focDE, and iucC) were also identified in the MDR isolates. The fimH gene was the most frequent (91.3%). Overall, 52 isolates from chicken (24 isolates, 96.0%), pig (16 isolates, 55.2%) and cattle (12 isolates, 80.0%) offals among MDR isolates were found to have some plasmid replicons. Frep (38 isolates) and FIB (27 isolates) replicons were more prevalent than other replicon types. The results suggest that edible offal can become a relevant reservoir of E. coli strains carrying various antimicrobial resistance and virulence genes. HIGHLIGHTS


2017 ◽  
Vol 80 (5) ◽  
pp. 734-739 ◽  
Author(s):  
Yung-Tsun Lo ◽  
Chia-Lan Wang ◽  
Bai-Hsung Chen ◽  
Chung-Wen Hu ◽  
Chung-Hsi Chou

ABSTRACT We tested 137 samples of domestic shucked oysters and 114 samples of imported oysters collected from traditional retail markets and supermarkets during 2010 and 2011 in Taiwan for the presence of Salmonella. We obtained a total of 91 Salmonella isolates, representing nine serotypes, from 80 of the domestic samples. We did not find any Salmonella in the imported oysters. The presence of Salmonella contamination tended to be specific to the area from which the oysters were harvested: the Dongshih area had a significantly higher contamination rate (68.8%) than the Budai (20.0%) and Wanggong (9.1%) areas. In addition, the rate of Salmonella contamination was higher in oysters that were packed or sold with water (P < 0.05). The most commonly identified Salmonella serotypes were Saintpaul (26.4%), Newport (22.0%), and Infantis (13.2%). We screened the isolates for susceptibility to nine antimicrobials and compared them genetically by using PCR for the class 1 integron (int1), tetA, tetB, and blaPSE-1 genes. Eighteen isolates (19.8%) were resistant to at least one antimicrobial agent, and the most frequent resistances were those to tetracycline and oxytetracycline (n = 12, 14.3%).We detected the antimicrobial resistance genes int1, tetA, tetB, and blaPSE-1 in 16.5, 26.4, 6.6, and 22.0% of the isolates, respectively. Eleven of the 18 antimicrobial-resistant isolates contained one or two int1 cassettes, suggesting that the presence of int1 is highly correlated with antimicrobial resistance in Salmonella isolates from oysters. The consumption of oysters is increasing in Taiwan, and information related to Salmonella contamination in oysters is rather limited. Our results indicate that raw oyster consumption from retail markets in Taiwan is associated with a human health hazard owing to Salmonella, including multidrug-resistant Salmonella strains.


Author(s):  
Dominic Poulin-Laprade ◽  
Jean-Simon Brouard ◽  
Nathalie Gagnon ◽  
Annie Turcotte ◽  
Alexandra Langlois ◽  
...  

Pigs are major reservoirs of resistant Enterobacteriaceae that can reach humans through consumption of contaminated meat or vegetables grown in manure-fertilized soil. Samples were collected from sows during lactation and their piglets at five time points spanning the production cycle. Cefotaxime-resistant bacteria were quantified and isolated from feed, feces, manures and carcasses of pigs reared with penicillin-using or antibiotic-free husbandries. The isolates were characterized by antibiotic susceptibility testing, whole genome sequencing and conjugation assays. The extended spectrum β-lactamase (ESBL) phenotype was more frequent in isolates originating from antibiotic-free animals, while the bacteria isolated from penicillin-using animals were on average resistant to a greater number of antibiotics. The ESBL-encoding genes identified were blaCTX-M-1, blaCTX-M-15 and blaCMY-2 and they co-localised on plasmids with various genes encoding resistance to ß-lactams, co-trimoxazole, phenicols and tetracycline, all antibiotics used in pig production. Groups of genes conferring the observed resistance and the mobile elements disseminating multidrug resistance were determined. The observed resistance to ß-lactams was mainly due to the complementary actions of penicillin-binding proteins, an efflux pump and ß-lactamases. Most resistance determinants were shared by animals raised with or without antimicrobials. This suggests a key contribution of indigenous enterobacteria maternally transmitted along the sow lineage, regardless of antimicrobial use. It is unclear if the antimicrobial resistance observed in the enterobacteria populations of the commercial pig herds studied were present before the use of antibiotics, or the extent to which historical antimicrobial use exerted a selective pressure defining the resistant bacterial populations in farms using penicillin prophylaxis. Importance: Antimicrobial resistance is a global threat that needs to be fought on numerous fronts along the One Health continuum. Vast quantities of antimicrobials are used in agriculture to ensure animal welfare and productivity, and are arguably a driving force for the persistence of environmental and food-borne resistant bacteria. This study evaluated the impact of conventional, organic and other antibiotic-free husbandry practices on the frequency and nature of antimicrobial resistance genes and multidrug resistant enterobacteria. It provides knowledge about the relative contribution of specific resistance determinants to observed antibiotic resistance. It also showed the clear co-selection of genes coding for extended-spectrum beta-lactamases and genes coding for the resistance to antibiotics commonly used for prophylaxis or in curative treatments in pig operations.


Author(s):  
Hornel Koudokpon ◽  
Victorien Dougnon ◽  
Christelle Lougbegnon ◽  
Esther Deguenon ◽  
Wassiyath Mousse ◽  
...  

Background: The environment plays an important role in the dissemination of multidrug resistant bacteria, especially through the aquatic ecosystem, including hospital effluents, rivers, but also spring water and drinking water. This study aims to determine selected antimicrobial resistance genes in some aquatic matrices in southern Benin. Methods: Collected water samples were filtered through a membrane 0.22 µm thick. After filtration, the membrane was deposited on Muëller Hinton agar. Then the colonies resulting from this subculture were subjected to a microbiological examination by the conventional method. The antibiotic sensitivity test was carried out by the Kirby Bauer method according to the recommendations of the French Society of Microbiology. Resistance genes were looked for by PCR. Results: Of the 222 water samples collected, 265 bacterial strains were isolated, the majority of which were strains of Coagulase Negative Staphylococcus (CNS) with 37.74% (n = 100), followed by strains of Klebsiella pneumoniae (21.89%; n = 58), Escherichia coli (10.57%; n = 28). All isolated gram-negative bacilli strains are multidrug resistant with resistance of all strains to amoxicillin, ampicillin and amoxicillin + clavulanic acid. Of the 15 resistance genes searched in the genome of Gram-negative bacilli strains, 8 were detected, namely the TEM, SHV, CTX-M15, VIM, NDM, SUL1, SUL2 and AADA genes. Resistance of CNS strains to amoxicillin, oxacillin and cefoxitin was observed. The meca gene was detected in all CNS strains. The vanA and VanB genes were only detected in strains isolated from drinking water in sachets collected from producers and street sellers. Conclusion: These results show the dissemination of resistance genes in Benin and once again confirms the urgency of a global fight against antimicrobial resistance.


2021 ◽  
Vol 12 ◽  
Author(s):  
Zengyuan Liu ◽  
Yingqiu Liu ◽  
Wei Xi ◽  
Shuangshi Liu ◽  
Jia Liu ◽  
...  

The genomic context of the mcr-1 gene in Escherichia coli from animal feces has been widely reported. However, less is known about the mcr-1-carrying plasmid characteristics and other functional regions of Escherichia coli isolates from animal organs with lesions. The present study investigated the antimicrobial resistance, population structure, and genetic features of mcr-1-positive Escherichia coli strains isolated from animal organs with lesions. The antimicrobial susceptibility testing indicated that 24 mcr-1-positive Escherichia coli isolates were resistant to at least three or all antimicrobial categories. MLST analysis suggested that the dominant clone complexes (CC) were mainly CC156, CC448, and CC10. In addition, ST10596, a newly discovered sequence type in swine, failed to be classified. Meanwhile, the mcr-1 gene located on the different plasmids was successfully transferred to the recipients, and whole-genome sequencing indicated the mcr-1 gene was embedded in mcr-1-pap2 cassette but not flanked by ISApl1. The mcr-1 gene is located on the chromosome and embedded in Tn6330. Furthermore, NDM-5 was found on the IncX3-type plasmid of J-8. The virB6 and traI gene of type IV secretion system (T4SS) were truncated by IS2 and IS100 and located on the IncX4- and the IncHI2/HI2A/N-type plasmids, respectively. The multidrug-resistant (MDR) region of IncHI2/HI2A/N-type plasmids contained two class 1 integrons (In0, In640) and four composite transposons (Tn4352, Tn6010, cn_4692_IS26, cn_6354_IS26). Overall, 24 mcr-1-positive Escherichia coli isolates in our study showed MDR, or even extensively drug resistant (XDR), and exhibited population diversity. The T4SS gene truncation by the insertion sequence may affect the efficiency of plasmid conjugative transfer. Furthermore, the class 1 integrons and composite transposons in the MDR region of IncHI2/HI2A/n-type plasmid contributed to the multireplicon plasmid formation, the acquisition, and transfer of antimicrobial resistance genes (ARGs).


2016 ◽  
Vol 79 (4) ◽  
pp. 589-597 ◽  
Author(s):  
LILI LI ◽  
RIKKE HEIDEMANN OLSEN ◽  
LEI YE ◽  
HE YAN ◽  
QING NIE ◽  
...  

ABSTRACT The aim of this study was to investigate the phenotypic and genotypic antimicrobial resistance, integrons, and transferability of resistance markers in 243 aerobic bacteria recovered from pork at slaughter in the People's Republic of China. The organisms belonged to 22 genera of gram-negative bacteria (92.2%) and gram-positive bacteria (7.8%). High levels of resistance were detected to tetracycline, trimethoprim-sulfamethoxazole, and ampicillin (36.2 to 54.3%), and lower levels were detected to nitrofurantoin, cefotaxime, gentamicin, ciprofloxacin, and chloramphenicol (7.8 to 29.2%). Across species, genes conferring antimicrobial resistance were observed with the following frequencies: blaTEM, 40.7%; blaCMY-2, 15.2%; blaCTX-M, 11.5%; sul2, 27.2%; sul1, 14.4%; tet(A), 5.4%; tet(L), 5.4%; tet(M), 5.0%; tet(E), 3.7%; tet(C), 3.3%; tet(S), 2.5%; and tet(K), 0.8%. Various antimicrobial resistance genes were found in new carriers: blaTEM in Lactococcus garvieae, Myroides odoratimimus, Aeromonas hydrophila, Staphylococcus sciuri, Raoultella terrigena, Macrococcus caseolyticus, Acinetobacter ursingii, Sphingobacterium sp., and Oceanobacillus sp.; blaCMY-2 in Lactococcus lactis, Klebsiella oxytoca, Serratia marcescens, Acinetobacter baumannii, and Myroides phaeus; tet(L) in M. caseolyticus; sul1 in Vibrio cincinnatiensis; sul2 in Acinetobacter bereziniae, Acinetobacter johnsonii, and V. cincinnatiensis; and the class 1 integron and gene cassette aadA2 in V. cincinnatiensis. Approximately 6.6% of isolates contained class 1 integrons, and one isolate harbored class 2 integrons. Plasmid associated intI1 and androgen receptor–encoding genes were transferred into Escherichia coli J53 and E. coli DH5α by conjugation and transformation experiments, respectively. Our study highlights the importance of aerobic bacteria from pork as reservoirs for antimicrobial resistance genes and mobile genetic elements that can readily be transferred intra- and interspecies.


Author(s):  
Anil Poudel ◽  
Terri Hathcock ◽  
Patrick Butaye ◽  
Yuan Kang ◽  
Stuart Price ◽  
...  

Background: Antimicrobial resistance is rising globally at an alarming rate. While multiple active surveillance programs have been established to monitor the antimicrobial resistance, studies on the environmental link to antimicrobial spread are lacking. Methods: A total of 493 flies were trapped from a dairy unit, a dog kennel, a poultry farm, a beef cattle unit, an urban trash facility and an urban downtown area to isolate Escherichia coli, Klebsiella pneumoniae and Staphylococcus spp. for antimicrobial susceptibility testing and molecular characterization. Results: E. coli, K. pneumoniae and coagulase-negative Staphylococcus were recovered from 43.9%, 15.5% and 66.2% of the houseflies, and 26.0%, 19.2%, 37.0% of the blowflies, respectively. In total, 35.3% of flies were found to harbor antimicrobial-resistant bacteria and 9.0% contained multidrug-resistant isolates. Three Staphylococcus aureus isolates were recovered from blowflies while three extended spectrum beta lactamase (ESBL)-carrying E. coli and one ESBL-carrying K. pneumoniae were isolated from houseflies. Whole genome sequencing identified the antimicrobial resistance genes blaCMY-2 and blaCTXM-1 as ESBLs. Conclusion: Taken together, our data indicate that flies can be used as indicators for environmental contamination of antimicrobial resistance. More extensive studies are warranted to explore the sentinel role of flies for antimicrobial resistance.


2020 ◽  
Vol 96 (8) ◽  
Author(s):  
Saharuetai Jeamsripong ◽  
Winn Khant ◽  
Rungtip Chuanchuen

ABSTRACT A total of 594 Vibrio parahaemolyticus isolates from cultivated oysters (n = 361) and estuarine water (n = 233) were examined for antimicrobial resistance (AMR) phenotype and genotype and virulence genes. Four hundred forty isolates (74.1%) exhibited resistance to at least one antimicrobial agent and 13.5% of the isolates were multidrug-resistant strains. Most of the V. parahaemolyticus isolates were resistant to erythromycin (54.2%), followed by sulfamethoxazole (34.7%) and trimethoprim (27.9%). The most common resistance genes were qnr (77.8%), strB (27.4%) and tet(A) (22.1%), whereas blaTEM (0.8%) was rarely found. Four isolates (0.7%) from oysters (n = 2) and estuarine water (n = 2) were positive to tdh, whereas no trh-positive isolates were observed. Significantly positive associations among AMR genes were observed. The SXT elements and class 1, 2 and 3 integrons were absent in all isolates. The results indicated that V. parahaemolyticus isolates from oysters and estuarine water were potential reservoirs of resistance determinants in the environment. This increasing threat of resistant bacteria in the environment potentially affects human health. A ‘One Health’ approach involved in multidisciplinary collaborations must be implemented to effectively manage antimicrobial resistance.


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