Evaluation of Culture Top transport systems for assessing the bacterial diversity of microbiota by culturomics as compared to a routine transport system

2021 ◽  
Vol 70 (10) ◽  
Author(s):  
Nadim Cassir ◽  
Souad Belkacemi ◽  
Mehdi Ballouche ◽  
Saber Khelaifia ◽  
Bernard La Scola

In recent years, metagenomics and then culturomics, which consists of the multiplication of media and culture conditions and the rapid identification of all bacterial colonies, have generated renewed interest in the human microbiota, and diseases associated with modifications in its composition in particular. The sample transport media included in diverse swab transport systems and the storage conditions are among the factors that influence the results of the culturomics. In this study, we compared the results of culturomics from paired skin, oral and rectal swabs from intensive care unit (ICU) patients using Culture Top sample transport medium as compared to our routine one. From 152 clinical samples, we were able to isolate and identify 45 600 colonies, belonging to 338 different bacterial species. The transport system Culture Top identified 282 different bacterial species, while 244 were identified by our routine system. Of these, 188 different bacterial species were commonly identified using both transport systems, while 94 (27.8 %) and 56 (16.5 %) were only identified using Culture Top and our routine system, respectively (P<0.001), but there was no significant difference in bacterial diversity at the genus or phylum level, or in terms of their type of respiration and cell wall. In conclusion, the Culture Top transport system appears to be complementary to our routine system, although it seems slightly superior in terms of isolated bacterial species.

2016 ◽  
Vol 2 (5) ◽  
pp. e1600300 ◽  
Author(s):  
Ki Soo Park ◽  
Chen-Han Huang ◽  
Kyungheon Lee ◽  
Yeong-Eun Yoo ◽  
Cesar M. Castro ◽  
...  

Health care–associated infections (HAIs) and drug-resistant pathogens have become a major health care issue with millions of reported cases every year. Advanced diagnostics would allow clinicians to more quickly determine the most effective treatment, reduce the nonspecific use of broad-spectrum antimicrobials, and facilitate enrollment in new antibiotic treatments. We present a new integrated system, polarization anisotropy diagnostics (PAD), for rapid detection of HAI pathogens. The PAD uses changes of fluorescence anisotropy when detection probes recognize target bacterial nucleic acids. The technology is inherently robust against environmental noise and economically scalable for parallel measurements. The assay is fast (2 hours) and performed on-site in a single-tube format. When applied to clinical samples obtained from interventional procedures, the PAD determined the overall bacterial burden, differentiated HAI bacterial species, and identified drug resistance and virulence status. The PAD system holds promise as a powerful tool for near-patient, rapid HAI testing.


2021 ◽  
Vol 7 (6) ◽  
pp. 433
Author(s):  
Ahmad Ibrahim ◽  
Lucie Peyclit ◽  
Rim Abdallah ◽  
Saber Khelaifia ◽  
Amanda Chamieh ◽  
...  

Candida auris is an emerging multidrug-resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation are necessary for diagnosis and containing its spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection ranging between 101 and 102 CFU/mL. The 100% specificity of SCA (specific C. auris) medium is confirmed on a set of 135 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, allowing the latter to study its phenotypic profile.


2018 ◽  
Vol 2018 (1) ◽  
pp. 000748-000753
Author(s):  
Terry Bluck ◽  
Chris Smith ◽  
Paul Werbaneth

Abstract Physical Vapor Deposition (PVD) systems are widely used in the semiconductor fabrication industry, both for front-end applications in the wafer fab and for back-end applications at device packaging houses. In fan-out wafer level packaging (FOWLP), and in fan-out panel level packaging (FOPLP), sputter deposited Ti and Cu are the base on which electroplated copper Redistribution Layers (RDLs) are built. For these RDL barrier/seed layers, PVD cluster tools, wafer transport architectures that have been widely used since the mid-1980s, are the current Process of Record (POR) in advanced packaging; however, these tools typically operate in a regime where wafer transport is robot-limited to approximately 50 wafers per hour, which limits overall throughput and greatly influences Cost of Ownership (COO) for the sputter deposition step(s), because the central handling robot occupied with a transfer from the Ti PVD module to the Cu PVD module, for example, has no opportunity to be doing anything other than that specific transfer. Other wafer transport architectures are more efficient from a wafer handling perspective. In linear transport carrier-based PVD tools, wafers or panels passing through the system benefit from a mechanical transfer time budget that is considerably less than for a cluster tool. Transport time overhead per wafer on linear transport systems is quite low, and scheduler software optimization becomes less onerous too, as a result of the simpler wafer transport architecture. We analyzed the relative throughput of cluster and linear transport PVD tools for a typical FOWLP barrier/seed layer (1000Å Ti / 2000Å Cu) sputter deposition process, and present details here of how the time spent moving wafers to various processing chambers affects overall system productivity. In the case of the cluster tool architecture, with its central wafer handling robot, wafer throughputs are approximately 50 wafers per hour, while on the linear transport system wafer throughputs as high as 240 wafers per hour are possible. The significant difference in system throughputs greatly affects the relative Cost of Ownership (COO) per wafer processed, with the linear transport system returning COO results that are less than half those of the typical cluster PVD tool.


2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Aliaa ELshamy ◽  
Zainab Zakaria ◽  
Mahmoud M. Tolba ◽  
Nermeen Salah Eldin ◽  
Al-Taher Rabea ◽  
...  

The emergence of AmpC (pAmpC) β-lactamases conferring resistance to the third-generation cephalosporins has become a major clinical concern worldwide. In this study, we aimed to evaluate the expression of AmpC β-lactamase encoding gene among the pathogenic Gram-positive and Gram-negative resistant bacteria screened from clinical samples of Egyptian patients enrolled into El-Qasr El-Ainy Tertiary Hospital in Cairo, Egypt. A total of 153 bacterial isolates of the species Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterococcus faecium were isolated from patients diagnosed with urinary tract infection (UTI), respiratory tract infection (RTI), and wound infections. The total number of E. faecium isolates was 53, comprising 29 urine isolates, 5 sputum isolates, and 19 wound swab isolates, whereas the total number of P. aeruginosa isolates was 49, comprising 27 urine isolates, 7 sputum isolates, and 15 wound swab isolates, and that of the K. pneumoniae isolates was 51, comprising 20 urine isolates, 25 sputum isolates, and 6 wound swab isolates. Our results indicated that there is no significant difference in the expression of AmpC β-lactamase gene among the tested bacterial species with respect to the type of infection and/or clinical specimen. However, the expression patterns of AmpC β-lactamase gene markedly differed according to the antibacterial resistance characteristics of the tested isolates.


2020 ◽  
Vol 2020 (10) ◽  
Author(s):  
Mohammed AlMogbel ◽  
Mohammed AlBolbol ◽  
Noura Elkhizzi ◽  
Hisham AlAjlan ◽  
John Philip Hays ◽  
...  

Abstract Nocardia cyriacigeorgica (N. cyriacigeorgica) is most frequently associated with human infections, including chronic bronchitis, pulmonary disease and brain abscesses. In general, N. cyriacigeorgica causes infections in immunocompromised individuals and has been reported in clinical samples worldwide. However, the isolation and speciation of N. cyriacigeorgica in the routine diagnostic microbiology laboratory are complicated and time consuming. Recent mass spectrometry techniques such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-TOF-MS) have been successfully integrated into many routine diagnostic microbiology laboratories, allowing for the rapid, accurate and simple identification and speciation of many different microorganisms, including difficult-to-identify bacterial species. Here, we present a case report of a 65-year-old female patient from the neurology ward of Prince Sultan Military Medical City in Riyadh, Saudi Arabia, who was infected with N. cyriacigeorgica. The bacterium was successfully identified by MALDI-TOF-MS, with species identification subsequently confirmed by sequence analysis of the 16S ribosomal RNA.


2000 ◽  
Vol 38 (4) ◽  
pp. 1672-1675 ◽  
Author(s):  
Lee-Jene Teng ◽  
Po-Ren Hsueh ◽  
Jui-Chang Tsai ◽  
Feng-Lin Chiang ◽  
Ching-Yi Chen ◽  
...  

Bacteroides thetaiotaomicron is the second most frequently encountered species of the anaerobes isolated from clinical specimens. We developed a PCR-based assay for the rapid identification of B. thetaiotaomicron. Specific primers were based on shared amplicons of about 1.2 kb generated from B. thetaiotaomicron by randomly amplified polymorphic DNA. This 1.2-kb fragment was sequenced and then used to design a set of PCR amplification primers. This PCR generated an amplification product of 721 bp, which was unique to all 65 isolates of B. thetaiotaomicron tested. There was no amplification with isolates of other bacterial species. Restriction enzyme digestion of the amplification product and dot blot hybridization further verified the specificity of the assay. These results suggest that this PCR assay targets a nucleotide sequence that is strongly conserved in B. thetaiotaomicron. This simple and rapid PCR assay provides a rapid and accurate method for identification of B. thetaiotaomicron and shows promise for the detection of B. thetaiotaomicron in clinical samples.


2018 ◽  
Vol 101 (4) ◽  
pp. 1135-1143 ◽  
Author(s):  
Irshad M Sulaiman ◽  
Pratik Banerjee ◽  
Ying-Hsin Hsieh ◽  
Nancy Miranda ◽  
Steven Simpson ◽  
...  

Abstract Staphylococcus spp. is considered as one of the most common human-pathogenic bacteria, causing illnesses ranging from nonthreatening skin infections to lethal diseases, including sepsis, pneumonia, bloodstream infections, and food poisoning. The emergence of methicillin-resistant Staphylococcus aureus strains has increased morbidity and mortality and resulted in a major healthcare burden worldwide. Single and multilocus sequence typing have been extensively used in the identification of Staphylococcus species. Nevertheless, these assays are relatively time-consuming and require high-quality DNA. Matrix-assisted laser desorption ionization-time-of-flight has been used recently for the rapid identification of several bacterial species. In this study, we have examined 47 Staphylococcus isolates recovered from food, environment, clinical samples, cosmetic products, and a medical device and 3 American Type Culture Collection Staphylococcus reference isolates using bioMérieux VITEK MS and VITEK 2 systems to determine isolate identity. Sequencing of the 16S ribosomal RNA gene was performed to confirm and compare the species identification data generated by VITEK 2 and VITEK MS systems. Although the VITEK 2 system could not identify one of the isolates, VITEK MS identified all 50 Staphylococcus spp. isolates tested. Results of this study clearly suggest that VITEK MS can be used in the rapid identification of Staphylococcus isolates of public health importance.


Author(s):  
Ahmad Ibrahim ◽  
Lucie Peyclit ◽  
Rim Abdallah ◽  
Saber Khelaifia ◽  
Amanda Chamieh ◽  
...  

Candida auris is an emerging multidrug resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation is necessary for its diagnosis and containing spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection of 102 CFU/ml. The 100% specificity of SCA (Specific C. auris) medium is confirmed on a set of 134 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, and allows studying its phenotypic profile.


2019 ◽  
Vol 31 (1) ◽  
pp. 48-51
Author(s):  
Suha S Hassan ◽  
Nidhal H. Ghaib ◽  
Batool H Al-Ghurabi

Background: The microorganisms can impend the life of health care professional and particularly the dental practitioners. They can be transmitted by different ways like airborne and droplet transmission. The current study was carried out to identify whether the arch wires that received from the manufactures are free from microbial contamination and to determine the bacterial species attached to the arch wires. Materials and Methods: This study involved eighty samples, consisted of two types of arch wires (nitinol and stainless-steel) from four companies (3M, G&H, Jiscop, OrthoTechnology). These wires inserted in a plane tube that contains 10 -ml of (Tris [tris(hydroxymethyl)aminomethane] and EDTA (ethylenediaminetetraacetic acid) tris-EDTA and brain heart infusion (BHI) broth. A 0.1 ml was withdrawn from the tube and spread on agar plates. The control groups consist of 16 plane tube (8 tubes with tris-EDTA and other 8 tubes with (BHI). Results: Microbial sampling yielded growth from 5 of the 80 arch wires. The predominant bacteria that isolated were Bacillus spp. No growth was recovered from 75 of the samples and from controls. The bacteria were isolated by BHI reagent and no growth was observed by tris-EDTA reagent with statistically significant difference (P<0.05). The Bacillus spp. found only in the G&H and Jiscop companies, however, no statistically significant difference was found among them (P>0.05). With regard to the presence and distribution of bacteria according to the types of wires, the present results clarified that cases of contamination with Bacillus spp. were found in the nitinol arch wires with statistically significant difference (P<0.05). Conclusions: The results of the current study revealed low count of bacterial contamination in the two types of companies (G&H and Jiscop). Not all materials that received from the manufactures are free from contamination and an effective sterilization regimen is needed to avoid cross-contamination.


2018 ◽  
Vol 41 (3) ◽  
pp. 255-264 ◽  
Author(s):  
J. Abraham Pérez-Pérez ◽  
David Espinosa-Victoria ◽  
Hilda V. Silva-Rojas ◽  
Lucía López-Reyes

Bacteria are an unavoidable component of the natural earthworm diet; thus, bacterial diversity in the earthworm gut is directly linked to decomposition of organic matter and development of the surrounding plants. The aim of this research was to isolate and to identify biochemically and molecularly the culturable bacterial microbiota of the digestive tract of Eisenia foetida. Earthworms were sourced from Instituto de Reconversión Productiva y Bioenergética (IRBIO) and Colegio de Postgraduados (COLPOS), México. Bacterial isolation was carried out on plates of Brain Heart Infusion (BHI) culture medium. Fifty six and 44 bacterial isolates were obtained from IRBIO and COLPOS, respectively. The population was composed of 44 Gram-negative and 56 Gram-positive isolates. Over 50 % of the bacterial isolates were rod-shaped cells. The 16S rRNA gene was sequenced and nine genera were identified in worms from IRBIO (Bacillus, Paenibacillus, Solibacillus, Staphylococcus, Arthrobacter, Pantoea, Stenotrophomonas, Acinetobacter and Aeromonas) and six in worms from COLPOS (Bacillus, Paenibacillus, Stenotrophomonas, Staphylococcus, Acinetobacter and Aeromonas). Bacillus was the predominant genus, with eight and six species in the oligochaetes from IRBIO and COLPOS, respectively. The most represented bacteria in the worms from both sites were Bacillus sp. and B. subtilis. The predominance of Bacillus was probably due to spore formation, a reproductive strategy that ensures survival and dispersion in the soil and oligochaetes digestive tract. The gut of E. foetida not only harbored bacterial species of agronomic importance but also species potentially pathogenic for humans (Staphylococcus warneri, Pantoea agglomerans and Stentrophomonas sp.). The larger bacterial diversity in worms from IRBIO could be due to their feeding on cattle manure, which is a rich source of bacteria.


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