Impact of Freezing on the Future Utility of Archived Surveillance Culture Specimens

2007 ◽  
Vol 28 (7) ◽  
pp. 886-888 ◽  
Author(s):  
Heather P. Green ◽  
Judith A. Johnson ◽  
Jon P. Furuno ◽  
Sandra M. Strauss ◽  
Eli N. Perencevich ◽  
...  

The ability to recover bacteria from frozen culture specimens has important implications. The purpose of this study was to validate the utility of frozen specimens for recovery of several gram-positive and gram-negative bacterial species by culture. Results demonstrate that 98% of 250 bacterial isolates identified on initial culture were subsequently recovered by culture of frozen specimens after a median storage period of 564 days.

2007 ◽  
Vol 9 (5) ◽  
pp. 404-410 ◽  
Author(s):  
Martina Greiner ◽  
Georg Wolf ◽  
Katrin Hartmann

Bacterial blood culture results of 292 privately owned cats presented to the Clinic for Small Animal Medicine, Ludwig Maximilian University Munich with signs of sepsis were evaluated retrospectively. Of the blood cultures, 23% were positive. In 88%, a single bacterial species was isolated. Of all bacterial isolates, 45% were Gram-positive, 43% were Gram-negative, and 12% were obligate anaerobes. The most frequently isolated bacteria were Enterobacteriaceae, obligate anaerobic species, Staphylococcus species and Streptococcus species. Of the cats with positive blood cultures, 32% were pretreated with antibiotics. Of all bacterial isolates, 77% were susceptible to enrofloxacin, 69% to chloramphenicol, 67% to gentamicin, and 64% to amoxycillin clavulanic acid. Only enrofloxacin reached an in vitro efficacy of more than 70% against Gram-positive and more than 74% against Gram-negative bacteria.


2021 ◽  
Vol 2 (2) ◽  
pp. 229-235
Author(s):  
Iqra Arooj ◽  
Alishba Sehar ◽  
Asghar Javaid

Prevalence and multidrug resistance among bacteria in catheter-associated urinary tract infections (CAUTIs) has been on the rise in recent times. Hence, the prevalence rate and antibacterial susceptibility of bacteria in CAUTIs in ICU patients was evaluated. A total of 120 patients admitted to the ICU of Nishtar Hospital, Multan, were recruited for this study. Both gram-positive and gram-negative bacterial isolates were characterized based on biochemical tests including catalase test, oxidase test, indole test, TSI test, citrate test, coagulase test and growth on 6.5% NaCl agar. The prevalence of bacterial species was Escherichia coli (32%), Staphylococcus aureus (26%), Pseudomonas spp. (18%), Proteus spp. (14%) and Enterococcus spp. (2%). A considerable degree of resistance against commonly prescribed antibiotics was observed. Gram negative bacteria showed resistance to ciprofloxacin, piperacillin-tazobactam and amikacin as well as susceptibility to imipenem, tigecycline and polymixin. Gram positive bacteria showed resistance to antibiotics such as piperacillin-tazobactam, ampicillin, gentamicin, oxacillin and ceftazidime suggesting the ineffectiveness of these antibiotics for treating bacterial infections among CAUTI patients and demonstrating the latest trends in antimicrobial drug resistance profile in local population.


2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S136-S136
Author(s):  
M Abdelmonem ◽  
A Gad AlKarim ◽  
S Eissa ◽  
A Boraik ◽  
M Shedid

Abstract Introduction/Objective Bacteremia is one of the major causes of life-threatening complications in patients with cancer. Significant changes in the spectrum of microorganisms isolated from blood culture BC have been reported in cancer patients over the past years. This study aimed to determine the predominant bacterial species causing bacteremia among febrile neutropenic FN cancer patients at the National Cancer Institute in Egypt (NCI). Methods A total of 300 BC collected from 300 FN cancer patients at NCI, Cairo. All cases were in patients with a mean age of 51 years, 158 patients were male (53%) while 142 patients were females (47%). BC was collected for microbiological investigations. Identification of the isolated organisms by the cultural characters (Morphological of bacterial isolates, Gram stain reaction, motility test, and biochemical tests) for each organism using standard semi- automated techniques. Results 68 (22.6%) BC were positive while 232 (77.4%) BC were negative. Gram-negative bacteria isolated and identified in 11 blood cultures (16.17%), while gram-positive isolates identified in 57 BC (83.8%). Among the Gram- negative organisms, 4 (5.8%) were Pseudomonas aeruginosa, 4 (5.8%) were E. coli, 1 (1.5%) was Klebssila pneumoni, 1 (1.5%) was Acintobacter and 1 (1.5%) was Citrobacter frenudiri. Among the Gram-positive organisms, Coagulase-negative Staphylococci CNS were most predominant in most cases 35 (61.4%). 7 (12%) were S. aureus, 5 (8%) were S. epidermises, 5 (8%) were Streptococcus spp., 1 (1.5%) were Listeria spp., 4 (5.88%) Achromobacter spp., 4 (5.88%) were Gram-Positive Cocci and 1(1.5%) Micrococcus spp. The study of R-factor in all positive BC showed the resistant bacterial isolates to the commonly used antimicrobial agents, especially to ampicillin and penicillin. Conclusion This study showed that patients with febrile neutropenia are vulnerable to developing bacteremia. the prevalence rate of bacteremia in post-chemotherapy FN in our center is relatively high compared to the national rate. Multidrug-resistant are the main cause of bacteremia in febrile cancer patients in Egypt. There is a need for ongoing antimicrobial surveillance to guide antimicrobial therapy and support the development of infection control programs in Egypt


Diagnostics ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 349
Author(s):  
Sien Ombelet ◽  
Alessandra Natale ◽  
Jean-Baptiste Ronat ◽  
Olivier Vandenberg ◽  
Liselotte Hardy ◽  
...  

Bacterial identification is challenging in low-resource settings (LRS). We evaluated the MicroScan identification panels (Beckman Coulter, Brea, CA, USA) as part of Médecins Sans Frontières’ Mini-lab Project. The MicroScan Dried Overnight Positive ID Type 3 (PID3) panels for Gram-positive organisms and Dried Overnight Negative ID Type 2 (NID2) panels for Gram-negative organisms were assessed with 367 clinical isolates from LRS. Robustness was studied by inoculating Gram-negative species on the Gram-positive panel and vice versa. The ease of use of the panels and readability of the instructions for use (IFU) were evaluated. Of species represented in the MicroScan database, 94.6% (185/195) of Gram-negative and 85.9% (110/128) of Gram-positive isolates were correctly identified up to species level. Of species not represented in the database (e.g., Streptococcus suis and Bacillus spp.), 53.1% out of 49 isolates were incorrectly identified as non-related bacterial species. Testing of Gram-positive isolates on Gram-negative panels and vice versa (n = 144) resulted in incorrect identifications for 38.2% of tested isolates. The readability level of the IFU was considered too high for LRS. Inoculation of the panels was favorably evaluated, whereas the visual reading of the panels was considered error-prone. In conclusion, the accuracy of the MicroScan identification panels was excellent for Gram-negative species and good for Gram-positive species. Improvements in stability, robustness, and ease of use have been identified to assure adaptation to LRS constraints.


2014 ◽  
Vol 11 (1) ◽  
pp. 66-70 ◽  
Author(s):  
S Shrestha ◽  
NC Shrestha ◽  
S Dongol Singh ◽  
RPB Shrestha ◽  
S Kayestha ◽  
...  

Background Neonatal sepsis is one of the major causes of morbidity and mortality among the newborns in the developing world. Objectives To determine the common bacterial isolates causing sepsis in neonatal intensive care unit and its antibiotic susceptibility pattern. Methods A one year discriptive prospective study was conducted in neonatal intensive care unit to analyse the results of blood culture and to look into the sensitivity of the commonly used antibiotics. Results The blood culture yield by conventional method was 44.13% with nosocomial sepsis accounting for 10.79%. 84.08% were culture proven early onset sepsis and 15.95% were late onset sepsis. Klebsiella infection was the commonest organism isolated in early, late and nosocomial sepsis but statistically not significant. Gram positive organisms were 39.36% in which Staphylococcus aureus was the leading microorganism followed by coagulase negative staphylococcus areus. Gram negative organisms were 60.64% amongst them Klebsiella was the most often encountered followed by Pseudomonas. The most common organism Klebsiella was 87.5% and 78.3% resistance to ampicillin and gentamycin respectively. Among gram negative isolates 87.5% and 77.2% were resistance to ampicillin and gentamycin respectively. Among gram positive isolates 58.5% and 31.5% resistance were noted to ampicillin and gentamycin respectively. Resistance to cefotaxim to gram negative and gram positive isolates were 87.34% and 59.35% respectively. Conclusion Klebsiella is most common organism which is almost resistance to first line antibiotics. Resistance to both gram negative and gram positive isolates among firstline antibiotics and even with cefotaxim is emerging and is a major concern in neonatal intensive care unit. DOI: http://dx.doi.org/10.3126/kumj.v11i1.11030 Kathmandu University Medical Journal Vol.11(1) 2013: 66-70


Micromachines ◽  
2018 ◽  
Vol 9 (8) ◽  
pp. 367 ◽  
Author(s):  
Yuguang Liu ◽  
Dirk Schulze-Makuch ◽  
Jean-Pierre de Vera ◽  
Charles Cockell ◽  
Thomas Leya ◽  
...  

Single-cell sequencing is a powerful technology that provides the capability of analyzing a single cell within a population. This technology is mostly coupled with microfluidic systems for controlled cell manipulation and precise fluid handling to shed light on the genomes of a wide range of cells. So far, single-cell sequencing has been focused mostly on human cells due to the ease of lysing the cells for genome amplification. The major challenges that bacterial species pose to genome amplification from single cells include the rigid bacterial cell walls and the need for an effective lysis protocol compatible with microfluidic platforms. In this work, we present a lysis protocol that can be used to extract genomic DNA from both gram-positive and gram-negative species without interfering with the amplification chemistry. Corynebacterium glutamicum was chosen as a typical gram-positive model and Nostoc sp. as a gram-negative model due to major challenges reported in previous studies. Our protocol is based on thermal and chemical lysis. We consider 80% of single-cell replicates that lead to >5 ng DNA after amplification as successful attempts. The protocol was directly applied to Gloeocapsa sp. and the single cells of the eukaryotic Sphaerocystis sp. and achieved a 100% success rate.


Wound infection is a major problem in hospitals in developing countries. Wound infection causes morbidity and prolonged hospital stay thus this prospective study was conducted for a period of seven months (January 2019 to July 2019). A total of 217 specimens (wound swabs and pus exudates) from wound infected patients in a Tertiary Care Hospital in Bangladesh. A retrospective study of the microbiological evaluation was done by cultural growth as well as Gram staining and biochemical examination to identify the bacterial isolates. Finally, the antimicrobial vulnerability testing was performed by Kirby-Bauer disc diffusion conventional method. A total of 295 samples were tested. Out of which 217 (73.5%) were found culture positive. E. coli was the most predominant gram-negative isolates whereas Staphylococcus aureus and Coagulase-negative Staphylococcus were the most commonly isolated gram-positive organisms. Antimicrobial sensitivity profile of bacterial isolates revealed imipenem, meropenem, amikacin, and nitrofurantoin to be the most effective antimicrobials against gram-negative isolates, whereas imipenem, meropenem, amikacin, nitrofurantoin, amoxiclav, and gentamicin were the most effective drugs against gram-positive isolates. The result of this examination contributes to the identification of basic causative microbes involved in wound infection and findings of antibiotic susceptibility patterns can be helpful for primary care physicians to optimize the treatment modalities, articulate policies for empiric antimicrobial therapy, and to minimize the rate of infection among wound infected patients.


2017 ◽  
Vol 43 (2) ◽  
pp. 173-180
Author(s):  
Md Abdul Karim ◽  
Nasrin Sultana

Present study was conducted to determine the microbiological status of water from dispensers in different roadside hotel and restaurants of Dhaka city. Samples were collected from seven hotel and restaurants. Aerobic heterotrophic bacterial count ranged between 1.5 × 10 and 8.8 × 103 cfu/ml. Enteric and related bacterial abundance in MacConkey, SS and Cetrimide agar media ranged from 0 to 4.9 × 106, 0 to 2.1 × 105 and 0 to 1.2 × 106cfu/ml, respectively. In total, 28 bacterial isolates were obtained during the study period. Among them, 15 were heterotrophic isolates and 13 were enteric and related bacteria. Among 15 aerobic heterotrophic isolates, 11 were gram positive and five were gram negative. Out of 11 gram positive isolates, 7 belonged to the genus Bacillus viz. B. circulans, B. subtilis, B. stearothermophilus, B. brevis and B. cereus and one to coccus viz. Micrococcus roseus. The other gram positive species were Kurtia gibsonii, Auriobacterium liguefaciens and Curtobacterium luteum. Four gram negative isolates were Neisseria elongate sub. spp. glycolytica, Plesiomonas shigelloides, Pseudomonas fluorescens biovar 1, Pseudomonas aeruginosa. All 13 enteric and related isolates were gram negative, short rod; and non-spore formers and belonged to the genera Escherichia, Klebsiella, Shigella and Pseudomonas. Among all isolates, two were resistant and six were susceptible to all five antibiotics. Asiat. Soc. Bangladesh, Sci. 43(2): 173-180, December 2017


2020 ◽  
Vol 13 (10) ◽  
pp. 2243-2251
Author(s):  
Azhar G. Shalaby ◽  
Neveen R. Bakry ◽  
Abeer A. E. Mohamed ◽  
Ashraf A. Khalil

Background and Aim: Flinders Technology Associates (FTA) cards simplify sample storage, transport, and extraction by reducing cost and time for diagnosis. This study evaluated the FTA suitability for safe transport and storage of Gram-positive and Gram-negative bacterial cells of animal origin on its liquid culture form and from organ impression smears (tissues) under the same routine condition of microbiological laboratory along with detecting their nucleic acid over different storage conditions. Materials and Methods: Increase in bacterial count from 104 to 107 (colony-forming units/mL) of 78 isolates representing seven bacterial species was applied onto cards. FTA cards were grouped and inoculated by these bacteria and then stored at different conditions of 24-27°C, 4°C, and –20°C for 24 h, for 2 weeks, for 1 and 3 month storage, respectively. Bacteriological examination was done, after which bacterial DNA was identified using specific primers for each bacterial type and detected by polymerase chain reaction (PCR). Results: The total percentage of recovered bacteria from FTA cards was 66.7% at 24-27–C for 24 h, the detection limit was 100% in Gram-positive species, while it was 57.4% in Gram-negative ones. Regarding viable cell detection from organ impression smears, it was successful under the previous conditions. No live bacterial cells were observed by bacteriological isolation rather than only at 24-27°C for 24 h storage. All bacterial DNA were sufficiently confirmed by the PCR technique at different conditions. Conclusion: Overall, the FTA card method was observed to be a valid tool for nucleic acid purification for bacteria of animal origin in the form of culture or organ smears regardless of its Gram type and is used for a short time only 24 h for storage and transport of live bacteria specifically Gram-positive type. Moreover, the bacterial nucleic acid was intact after storage in –20°C for 3 months and was PCR amplifiable.


Parasitology ◽  
2019 ◽  
Vol 147 (1) ◽  
pp. 29-38
Author(s):  
Rory Gough ◽  
Joel Barratt ◽  
Damien Stark ◽  
John Ellis

AbstractThe presence of bacterial DNA in Dientamoeba fragilis DNA extracts from culture poses a substantial challenge to sequencing the D. fragilis genome. However, elimination of bacteria from D. fragilis cultures has proven difficult in the past, presumably due to its dependence on some unknown prokaryote/s. This study explored options for removal of bacteria from D. fragilis cultures and for the generation of genome sequence data from D. fragilis. DNA was extracted from human faecal samples and xenic D. fragilis cultures. Extracts were subjected to 16S ribosomal DNA bacterial diversity profiling. Xenic D. fragilis cultures were then subject to antibiotic treatment regimens that systematically removed bacterial species depending on their membrane structure (Gram-positive or Gram-negative) and aerobic requirements. The impact of these treatments on cultures was assessed by 16S amplicon sequencing. Prior to antibiotic treatment, the cultures were dominated by Gram-negative bacteria. Addition of meropenem to cultures eliminated anaerobic Gram-negative bacteria, but it also led to protozoan death after 5 days incubation. The seeding of meropenem resistant Klebsiella pneumoniae strain KPC-2 into cultures before treatment by meropenem prevented death of D. fragilis cells beyond this 5 day period, suggesting that one or more species of Gram-negative bacteria may be an essential nutritional requirement for D. fragilis. Gram-positive cells were completely eliminated using vancomycin without affecting trophozoite growth. Finally, this study shows that genome sequencing of D. fragilis is feasible following bacterial elimination from cultures as the result of the major advances occurring in bioinformatics. We provide evidence on this fact by successfully sequencing the D. fragilis 28S large ribosomal DNA subunit gene using culture-derived DNA.


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