ISOLATION METHODS FOR MOLECULAR DETECTION AND ANTIBIOTIC RESISTANCE PATTERN OF CAMPYLOBACTER SPP IN LAYER CHICKENS

This study was conducted to compare two culture methods for the isolation of Campylobacter spp from commercial layer chickens and subsequently confirmed by Polymerase Chain Reaction assays (PCR). Furthermore, the antimicrobial resistance profiles of PCR positive Campylobacter isolates were determined.Cloacal swab samples (550) from chickens randomly selected from five poultry farms in the four geographical zones in Ogun State were cultured for Campylobacter using modified charcoal Cefoperazone deoxycholate agar (MCCDA) and an improved culture method involving Preston broth pre-enrichment and subsequent subculture on Mueller Hinton agar with Campylobacter growth supplements. Putative isolates were later confirmed by PCR assay and sequencing analysis.Other isolates that grew on MCCDA and confirmed by sequencing analysis are Enterococcus faecalis, Escherichis coli, Comamonas kerstli and Pseudomonas aeroginusa . The antibiotic resistant profile of all the isolates were evaluated genotypically for resistance genes to tetracyclines (tetO), multiclasses (cmeB), aminoglycosides (aphA-3-1) and β-lactams (Blaoxa-61) using multiplex PCR (mPCR), and phenotypically for chlortetracycline, tylosin, streptomycin, ciprofloxacin and erythromycin resistance by microbroth dilution method which correspond to the antibiotic resistance genes. The apparent prevalence of Campylobacter was 16.8% by MCCDA while none of the isolates was positive to PCR. Meanwhile, prevalence rate of 26% was obtained using Preston broth pre-enrichment and Mueller Hinton agar with Campylobacter growth supplements, of which 11/50 (22%) of the isolates was confirmed positive by PCR. Genotypic characterization of PCR positive isolates showed 10/11(90%) were C. coli, 1/11(10%) other Campylobacter species and 0% C. jejuni. All the isolates carried both tetO and cmeB resistant genes. The results of minimum inhibitory concentration presented all PCR positive isolates had resistance of 10/10(100%), 9/10(90%), 6/10(60%), 9/10(90%), and 8/10(80%) to tetracycline, ciprofloxacin, erythromycin, spectinomycin and tylosin respectively. In addition, all isolates carried multiple resistance to most antibiotics tested which are commonly used in poultry practice in Nigeria. Campylobacter spp in the study areas showed diverse genotypic characteristics, and gene mediated multidrug resistance.

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Molecular detection of extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates of chicken origin from East Java, Indonesia

Veterinary World ◽

10.14202/vetworld.2019.578-583 ◽

2019 ◽

Vol 12 (4) ◽

pp. 578-583 ◽

Cited By ~ 1

Author(s):

Meutia Hayati ◽

Agustin Indrawati ◽

Ni Luh Putu Ika Mayasari ◽

Istiyaningsih Istiyaningsih ◽

Neneng Atikah

Keyword(s):

Antibiotic Resistance ◽

Klebsiella Pneumoniae ◽

Resistance Gene ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Resistance Pattern ◽

Dilution Method ◽

Agar Dilution Method ◽

Resistance Level ◽

Extended Spectrum

Background and Aim: Klebsiella pneumoniae is one of the respiratory disease agents in human and chicken. This bacterium is treated by antibiotic, but this treatment may trigger antibiotic resistance. Resistance gene in K. pneumoniae may be transferred to other bacteria. One of the known resistance genes is extended-spectrum β-lactamase (ESBL). This research aimed to study K. pneumoniae isolated from chicken farms in East Java, Indonesia, by observing the antibiotic resistance pattern and detect the presence of ESBL coding gene within the isolates. Materials and Methods: A total of 11 K. pneumoniae isolates were collected from 141 chicken cloacal swabs from two regencies in East Java. All isolates were identified using the polymerase chain reaction method. Antimicrobial susceptibility was determined by agar dilution method on identified isolates, which then processed for molecular characterization to detect ESBL coding gene within the K. pneumoniae isolates found. Results: The result of antibiotic sensitivity test in 11 isolates showed highest antibiotic resistance level toward ampicillin, amoxicillin, and oxytetracycline (100%, 100%, and 90.9%) and still sensitive to gentamicin. Resistance against colistin, doxycycline, ciprofloxacin, and enrofloxacin is varied by 90.9%, 54.5%, 27.3%, and 18.2%, respectively. All isolates of K. pneumoniae were classified as multidrug resistance (MDR) bacteria. Resistance gene analysis revealed the isolates harbored as blaSHV (9.1%), blaTEM (100%), and blaCTX-M (90.9%). Conclusion: All the bacterial isolates were classified as MDR bacteria and harbored two of the transmissible ESBL genes. The presence of antibiotic resistance genes in bacteria has the potential to spread its resistance properties.

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Screening of Punica granatum extract for antimicrobial activity against oral micro organisms

Journal of Ayurvedic and Herbal Medicine ◽

10.31254/jahm.2020.6211 ◽

2020 ◽

Vol 6 (2) ◽

pp. 73-77

Author(s):

Meera Avadhani ◽

◽

Meena Anand Kukkamalla ◽

Kishore G Bhat ◽

◽

...

Keyword(s):

Research Work ◽

Fusobacterium Nucleatum ◽

Punica Granatum ◽

Dilution Method ◽

Dental Patients ◽

Mueller Hinton ◽

Mueller Hinton Agar ◽

Serial Dilution Method ◽

Micro Organisms ◽

Dental Infections

Background and Objectives: A lot of research work in both dental and medical fields support the curative properties of pomegranate. Accordingly, it was decided to prepare a pomegranate mouthwash and evaluate it among Dental patients diagnosed with Chronic Gingivitis. The objective of the present invitro study is to assess the Minimum Inhibitory Concentration (MIC) of the commercially available pomegranate extract powder against few oral pathogenic microorganisms. Methodology: Serial dilution method using thioglycolate broth medium was used for anerobes like Streptococcus mutans, Fusobacterium nucleatum, Aggregatibacter actinomycetomcomitans, Prevotella intermedia and Mueller hinton agar mediated growth was used for aerobe like Staphylococcus aureus. Following which microdilution assay was performed and accordingly evaluated the MIC. Based on this report, the test rinse was prepared and further evaluated using the same methodology for both aerobes and anerobes. Results and Inference: It was observed from the MIC report for both aerobes and anerobes that at a concentration of 0.2% the formulated mouth rinse was effective against all the chosen organisms. The results of the study infer that products like mouthwash, dental gels etc made from this concentration could be possibly used for the control of dental infections.

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Antibiotic resistance patterns of Pseudomonas spp. isolated from faecal wastes in the environment and contaminated surface water

FEMS Microbiology Ecology ◽

10.1093/femsec/fiaa008 ◽

2020 ◽

Vol 96 (2) ◽

Cited By ~ 4

Author(s):

Mathilde Camiade ◽

Josselin Bodilis ◽

Naouel Chaftar ◽

Wassila Riah-Anglet ◽

Johan Gardères ◽

...

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Enteric Bacteria ◽

Resistance Pattern ◽

Pathogenic Species ◽

Pseudomonas Species ◽

Class 1 ◽

Resistance Patterns ◽

Faecal Bacteria ◽

Antibiotic Resistance Patterns

ABSTRACT The Pseudomonas genus, which includes environmental and pathogenic species, is known to present antibiotic resistances, and can receive resistance genes from multi-resistant enteric bacteria released into the environment via faecal rejects. This study was aimed to investigate the resistome of Pseudomonas populations that have been in contact with these faecal bacteria. Thus, faecal discharges originating from human or cattle were sampled (from 12 points and two sampling campaigns) and 41 Pseudomonas species identified (316 isolates studied). The resistance phenotype to 25 antibiotics was determined in all isolates, and we propose a specific antibiotic resistance pattern for 14 species (from 2 to 9 resistances). None showed resistance to aminoglycosides, tetracycline, or polymyxins. Four species carried a very low number of resistances, with none to β-lactams. Interestingly, we observed the absence of the transcriptional activator soxR gene in these four species. No plasmid transfer was highlighted by conjugation assays, and a few class 1 but no class 2 integrons were detected in strains that may have received resistance genes from Enterobacteria. These results imply that the contribution of the Pseudomonas genus to the resistome of an ecosystem first depends on the structure of the Pseudomonas populations, as they may have very different resistance profiles.

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Antimicrobial susceptibility, multilocus sequence typing, and virulence of listeria isolated from a slaughterhouse in Jiangsu, China

BMC Microbiology ◽

10.1186/s12866-021-02335-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Liting Wu ◽

Hongduo Bao ◽

Zhengquan Yang ◽

Tao He ◽

Yuan Tian ◽

...

Keyword(s):

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Antimicrobial Susceptibility ◽

Multilocus Sequence Typing ◽

Tetracycline Resistance ◽

Dilution Method ◽

Processing Plant ◽

Meat Processing ◽

Processing Plants

Abstract Background Listeria monocytogenes is one of the deadliest foodborne pathogens. The bacterium can tolerate severe environments through biofilm formation and antimicrobial resistance. This study aimed to investigate the antimicrobial susceptibility, resistance genes, virulence, and molecular epidemiology about Listeria from meat processing environments. Methods This study evaluated the antibiotic resistance and virulence of Listeria isolates from slaughtering and processing plants. All isolates were subjected to antimicrobial susceptibility testing using a standard microbroth dilution method. The harboring of resistant genes was identified by polymerase chain reaction. The multilocus sequence typing was used to determine the subtyping of the isolates and characterize possible routes of contamination from meat processing environments. The virulence of different STs of L. monocytogenes isolates was evaluated using a Caco-2 cell invasion assay. Results A total of 59 Listeria isolates were identified from 320 samples, including 37 L. monocytogenes isolates (62.71%). This study evaluated the virulence of L. monocytogenes and the antibiotic resistance of Listeria isolates from slaughtering and processing plants. The susceptibility of these 59 isolates against 8 antibiotics was analyzed, and the resistance levels to ceftazidime, ciprofloxacin, and lincomycin were as high as 98.31% (L. m 37; L. innocua 7; L. welshimeri 14), 96.61% (L. m 36; L. innocua 7; L. welshimeri 14), and 93.22% (L. m 35; L. innocua 7; L. welshimeri 13), respectively. More than 90% of the isolates were resistant to three to six antibiotics, indicating that Listeria isolated from meat processing environments had high antimicrobial resistance. Up to 60% of the isolates harbored the tetracycline-resistance genes tetA and tetM. The frequency of ermA, ermB, ermC, and aac(6′)-Ib was 16.95, 13.56, 15.25, and 6.78%, respectively. Notably, the resistant phenotype and genotype did not match exactly, suggesting that the mechanisms of antibiotic resistance of these isolates were likely related to the processing environment. Multilocus sequence typing (MLST) revealed that 59 Listeria isolates were grouped into 10 sequence types (STs). The dominant L. monocytogenes STs were ST5, ST9, and ST121 in the slaughtering and processing plant of Jiangsu province. Moreover, ST5 subtypes exhibited high invasion in Caco-2 cells compared with ST9 and ST121 cells. Conclusion The dominant L. monocytogenes ST5 persisted in the slaughtering and processing plant and had high antimicrobial resistance and invasion characteristics, illustrating a potential risk in food safety and human health.

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Antibiotic Resistance Pattern of Bacteroides Fragilis Isolated From Clinical and Gastrointestinal Specimens

10.21203/rs.3.rs-209897/v1 ◽

2021 ◽

Author(s):

Seyedesomaye Jasemi ◽

Mohammad Emaneini ◽

Zahra Ahmadinejad ◽

Mohammad Sadegh Fazeli ◽

Leonardo A. Sechi ◽

...

Keyword(s):

Antibiotic Resistance ◽

Bacteroides Fragilis ◽

Antibiotic Resistance Genes ◽

Penicillin G ◽

Clinical Samples ◽

Resistance Pattern ◽

Dilution Method ◽

Agar Dilution Method ◽

Gi Tract ◽

Antibiotic Resistance Pattern

Abstract Background: Bacteroides fragilis is a part of the normal gastrointestinal flora and the most prevalent anaerobic bacteria causes’ infection. It is highly resistant to antibiotics and contains abundant antibiotic resistance mechanisms. Methods: The antibiotic resistance pattern of 78 isolates of B. fragilis (56 strains from the gastrointestinal [GI] tract and 22 strains from clinical samples) was investigated using agar dilution method. The gene encoding Bacteroides fargilis toxin bft, and antibiotic resistance genes were targeted by PCR assay. Results: The highest rate of resistance was observed for penicillin G (100%) followed by tetracycline (74.4%), clindamycin (41%) and cefoxitin (38.5%). Only a single isolate showed resistance to imipenem which contained cfiA and IS1186 genes. All isolates were susceptible to metronidazole. Accordingly, tetQ (87.2%), cepA (73.1%) and ermF (64.1%) were the most abundant antibiotic-resistant genes identified in this study. MIC values for penicillin, cefoxitin and clindamycin were significantly different among isolates with the cepA, cfxA and ermF in compare with those lacking such genes. In addition, 22.7% and 17.8% of clinical and GI tract isolates had the bft gene, respectively. Conclusions: Therefore, it is of utmost importance to determine the antibiotic resistance patterns of B. fragilis periodically in different geographical areas to provide a suitable treatment profile for patients and to prevent improper antibiotic prescriptions.

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Phylum barrier and Escherichia coli intra-species phylogeny drive the acquisition of antibiotic-resistance genes

Microbial Genomics ◽

10.1099/mgen.0.000489 ◽

2021 ◽

Vol 7 (8) ◽

Author(s):

Marie Petitjean ◽

Bénédicte Condamine ◽

Charles Burdet ◽

Erick Denamur ◽

Etienne Ruppé

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Resistance Genes ◽

Type Species ◽

Antibiotic Resistance Genes ◽

Resistance Pattern ◽

Content Type ◽

E Coli ◽

Link Type ◽

Specific Distribution

Escherichia coli is a ubiquitous bacterium that has been widely exposed to antibiotics over the last 70 years. It has adapted by acquiring different antibiotic-resistance genes (ARGs), the census of which we aim to characterize here. To do so, we analysed 70 301 E. coli genomes obtained from the EnteroBase database and detected 1 027 651 ARGs using the AMRFinder, Mustard and ResfinderFG ARG databases. We observed a strong phylogroup and clonal lineage specific distribution of some ARGs, supporting the argument for epistasis between ARGs and the strain genetic background. However, each phylogroup had ARGs conferring a similar antibiotic class resistance pattern, indicating phenotypic adaptive convergence. The G+C content or the type of ARG was not associated with the frequency of the ARG in the database. In addition, we identified ARGs from anaerobic, non- Proteobacteria bacteria in four genomes of E. coli , supporting the hypothesis that the transfer between anaerobic bacteria and E. coli can spontaneously occur but remains exceptional. In conclusion, we showed that phylum barrier and intra-species phylogenetic history are major drivers of the acquisition of a resistome in E. coli .

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Evolution of Antibiotic Resistance in Enterobacter Spp Isolated at the Yaounde University Teaching Hospital from January 2008 to November 2019

Open Access Journal of Microbiology & Biotechnology ◽

10.23880/oajmb-16000191 ◽

2021 ◽

Vol 6 (2) ◽

pp. 1-6

Author(s):

Kamga HG

Keyword(s):

Antibiotic Resistance ◽

University Teaching ◽

Phenotypic Characterization ◽

Phenotypic Analysis ◽

Surveillance Strategy ◽

Adequate Treatment ◽

Antibiotic Resistance Profile ◽

Mueller Hinton ◽

Mueller Hinton Agar ◽

Synergy Test

Purpose: A study was conducted to evaluate the evolution of the resistance of Enterobacter spp to antibiotics during twelve years and to update the data. Method: A retro-prospective study was carried from January 2008 to November 2019. Data was extracted from the registers of the bacteriology laboratory and the strains from samples received from the different units of the YUTH. The study of the antibiotic resistance profile of these species and phenotypic analysis was carried out by the method of discs diffusion in Mueller-Hinton agar. Phenotypic characterization was carried out by synergy test and modified Hodge test. Findings: A total of 109 strains were isolated in our study. Enterobacter species showed high resistance with a peak in 2012 for cephalosporins, in 2011 for aminoglycosides, in 2018 for quinolones, in 2019 for carbapenems with the frequencies of 80%, 45%, 37% and 36,1% respectively. These species exhibited 30% resistance to colistin. The resistance peak to the majority of antibiotics between 2018 and 2019 reflects an increase of resistance. The Extended Spectrum  - Lactamases (ESBL) phenotype was the most represented with frequency of 32.4%. Unique contri bution to theory, practice and policy: To Update the data on the evolution of Enterobacter spp, which will help to establish a surveillance strategy in Cameroonand adapt an adequate treatment regimen.

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Correlation of Oxacillin MIC with mecAGene Carriage in Coagulase-Negative Staphylococci

Journal of Clinical Microbiology ◽

10.1128/jcm.38.2.752-754.2000 ◽

2000 ◽

Vol 38 (2) ◽

pp. 752-754 ◽

Cited By ~ 68

Author(s):

Zafar Hussain ◽

Luba Stoakes ◽

Viki Massey ◽

Deb Diagre ◽

Viivi Fitzgerald ◽

...

Keyword(s):

Staphylococcus Epidermidis ◽

Clinical Laboratory ◽

National Committee ◽

Dilution Method ◽

Agar Dilution Method ◽

Coagulase Negative Staphylococci ◽

Methicillin Resistant ◽

Mueller Hinton ◽

Mueller Hinton Agar ◽

Agar Dilution

The National Committee for Clinical Laboratory Standards has recently changed the oxacillin breakpoint from ≥4 mg/liter to ≥0.5 mg/liter to detect methicillin-resistant coagulase-negative staphylococci (CoNS) because the previous breakpoint lacked sensitivity. To determine the correlation between the new oxacillin breakpoint and the presence of themecA gene, 493 CoNS of 11 species were tested. The presence of the mecA gene was determined by PCR, and oxacillin susceptibility was determined by the agar dilution method with Mueller-Hinton agar containing 2% NaCl and oxacillin (0.125 to 4.0 mg/liter). The new breakpoint correctly classified all CoNS strains with mecA as methicillin resistant and strains ofStaphylococcus epidermidis, S. haemolyticus, and S. hominiswithout mecA as methicillin susceptible. The breakpoint of ≥0.5 mg/liter was not specific for S. cohnii, S. lugdunensis, S. saprophyticus, S. warneri, and S. xylosus, in that it categorized 70 of 74 strains of these species withoutmecA (94.6%) as methicillin resistant. The results of this study indicate that the new oxacillin breakpoint accurately identifies strains of CoNS with mecAbut is not specific for strains of certain species of CoNS withoutmecA.

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The Antibiotic Resistance Pattern and Prevalence of blaTEM, blaSHV, blaCTX-M, blaPSE-1, sipB/C, and cmlA/tetR Genes in Salmonella typhimurium Isolated from Children with Diarrhea in Tabriz, Iran

International Journal of Health and Life Sciences ◽

10.5812/ijhls.118523 ◽

2021 ◽

Vol In Press (In Press) ◽

Author(s):

Abolfazl Jafari Sales ◽

Sara Naebi ◽

Rozita Nasiri ◽

Hossein Bannazadeh-Baghi

Keyword(s):

Antibiotic Resistance ◽

Salmonella Typhimurium ◽

Resistance Genes ◽

Health Concern ◽

Resistance Pattern ◽

Cross Sectional ◽

Fecal Samples ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamases ◽

Synergy Test

Background: Salmonella gastroenteritis is a global health concern. Recently, increased resistance to Salmonella typhimurium has been reported in several countries. Objectives: The present study aimed to evaluate the prevalence of blaTEM, blaSHV, blaCTX-M, blaPSE-1, sipB/C, and cmlA/tetR genes in S. typhimurium isolates and determine their antibiotic resistance. Methods: This cross-sectional descriptive study was conducted on 110 fecal samples, which were collected from the patients referred to the hospitals and medical centers in Tabriz, Iran during eight months. After phenotypic identification, the antibiogram test and double-disc synergy test were performed on the isolates. Following that, the prevalence of resistance genes was evaluated using multiplex PCR and specific primers. Results: Out of 110 fecal samples, 26 samples (23.63%) were positive for S. typhimurium. The highest resistance of the isolates was against ceftazidime, cefotaxime, amikacin, and tetracycline (100%), and the lowest resistance was against imipenem (3.85%) and nalidixic acid (7.69%). In total, 15 S. typhimurium isolates (57.69%) were positive for the extended-spectrum beta-lactamases. In addition, the most common resistance genes in the isolates were cmlA/tetR (38.46%), blaTEM (34.61%), and blaCTX-M (26.92%). Four isolates (15.38%) carried sipB, three isolates (11.53%) contained blaSHV, and two isolates (7.69%) carried blaPSE-1. Conclusions: The obtained results indicated the high prevalence of antibiotic-resistant S. typhimurium. Therefore, the identification of resistance genes is an important strategy for identifying and counteracting antibiotic resistance.

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In vitro evalution of antibiotic resistance of Lactobacillus bulgaricus strains isolated from traditional dairy products

Czech Journal of Food Sciences ◽

10.17221/136/2018-cjfs ◽

2019 ◽

Vol 37 (No. 1) ◽

pp. 36-43

Author(s):

Huiling Guo ◽

Bilige Menghe

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Antibiotic Resistance Gene ◽

Antibiotic Resistance Genes ◽

Fermented Milk ◽

Lactobacillus Bulgaricus ◽

Dilution Method ◽

Resistance Transfer ◽

Traditional Dairy Products

The antimicrobial susceptibility of 20 Lb. bulgaricus isolates from traditional fermented milk-originated was assessed and then determined the ability to transfer antibiotic resistance genes to other bacteria. The minimum inhibitory concentration of each strain was determined using a standardized dilution method. All the tested strains were found to be susceptible to gentamicin, erythromycin, clindamycin, neomycin, tetracycline, linezolid, chloramphenicol, rifampicin, and quinupristin/dalfopristin, while their susceptibilities to kanamycin, ciprofloxacin, streptomycin, trimethoprim, ampicillin, and vancomycin varied. Polymerase chain reaction (PCR) was used to check whether specific antibiotic resistance genes were present in these Lb. bulgaricus. We detected the rpoB, erm(B), aadA, bla, cat and vanX. Finally, a filter mating assay was applied to investigate the transferability of these resistance markers; and we observe no antibiotic resistance transfer between bacteria. This work demonstrates a low risk of lateral transfer of the antibiotic resistance gene of Lb. bulgaricus.

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