Fundamental Techniques in Cell Culture

In partnership with Sigma-Aldrich, ECACC have produced a third edition of ‘Fundamental Techniques in Cell Culture’. This compact laboratory handbook provides a wealth of information from the sourcing of cell lines, safety and laboratory design to aspects of cryopreservation and quality control. A series of 10 detailed protocols are provided, which are routinely used in the ECACC laboratories along with detailed protocols from the European Bank for induced Pluripotent Stem Cells (EBiSC) on how to culture and freeze induced pluripotent cell (iPSC) lines.
REFERENCE:
Fundamental Techniques in Cell Culture, Laboratory Handbook -Third Edition

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Strain, disinfectant, concentration, and contact time quantitatively impact disinfectant efficacy

Transmission of healthcare-associated infections caused by antibiotic- and multi-drug resistant (MDR) pathogens (e.g. Methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa) are a major concern in patient care facilities. Disinfectant usage is critical to control and prevent pathogen transmission, yet the relationships among strain, disinfectant type, contact time, and concentration are not well-characterized. We hypothesized that there would be significant differences in disinfectant efficacy among clinically relevant strains under off-label disinfectant conditions, but there would be less no differences among at registered label use concentrations and contact times. The purpose of this study was to quantify the effect of disinfectant concentration and contact time on the bactericidal efficacy of clinically relevant strains of S. aureus and P. aeruginosa.
Accelerated hydrogen peroxide (AHP), quaternary ammonium compounds (Quat), and sodium hypochlorite were tested at label and reduced contact times and concentrations against four MDR P. aeruginosa strains and four MRSA strains. Quantitative EPA method MB-25-02 was used to measure disinfectant efficacy reported as log10 reduction.
Both off-label disinfectant concentrations and contact times significantly affected efficacy of all disinfectants tested. Bactericidal efficacy varied among MRSA and P. aeruginosa strains.
The quantitative disinfectant efficacy method used highlights the inter-strain variability that exists within a bacterial species. It also underscores the need for a disinfectant validation method that takes these variances into account.
REFERENCE:
West, Alyssa M et al. “Strain, disinfectant, concentration, and contact time quantitatively impact disinfectant efficacy” Antimicrobial resistance and infection control vol. 7 49. 3 Apr. 2018, doi:10.1186/s13756-018-0340-2

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Laboratory management of Crimean-Congo haemorrhagic fever virus infections

Background: Crimean-Congo haemorrhagic fever virus (CCHFV) is considered an emerging infectious disease threat in the European Union. Since 2000, the incidence and geographic range of confirmed CCHF cases have markedly increased, following changes in the distribution of its main vector, Hyalomma ticks. Aims: To review scientific literature and collect experts' opinion to analyse relevant aspects of the laboratory management of human CCHF cases and any exposed contacts, as well as identify areas for advancement of international collaborative preparedness and laboratory response plans. Methods: We conducted a literature review on CCHF molecular diagnostics through an online search. Further, we obtained expert opinions on the key laboratory aspects of CCHF diagnosis. Consulted experts were members of two European projects, EMERGE (Efficient response to highly dangerous and emerging pathogens at EU level) and EVD-LabNet (Emerging Viral Diseases-Expert Laboratory Network).Results: Consensus was reached on relevant and controversial aspects of CCHF disease with implications for laboratory management of human CCHF cases, including biosafety, diagnostic algorithm and advice to improve lab capabilities. Knowledge on the diffusion of CCHF can be obtained by promoting syndromic approach to infectious diseases diagnosis and by including CCHFV infection in the diagnostic algorithm of severe fevers of unknown origin. Conclusion: No effective vaccine and/or therapeutics are available at present so outbreak response relies on rapid identification and appropriate infection control measures. Frontline hospitals and reference laboratories have a crucial role in the response to a CCHF outbreak, which should integrate laboratory, clinical and public health responses.
REFERENCE:
Bartolini B, et al. Laboratory management of Crimean-Congo haemorrhagic fever virus infections: perspectives from two European networks. Euro Surveill. 2019 Jan;24(5). doi: 10.2807/1560-7917.ES.2019.24.5.1800093. PubMed PMID: 30722811.


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COMUNICADO AMEXBIO

La AMEXBIO emitió un comunicado sobre la nueva iniciativa de Ley de de Humanidades, Ciencias y Tecnologías. Pueden descargar el documento desde la página: www.amexbio.org




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Bactericidal efficacy of ready-to-use disinfectant Towelettes

Disinfectant products are often used on environmental surfaces (e.g. countertops, patient beds) and patient care equipment in healthcare facilities to help prevent the transmission of healthcare-associated infections. Ready-to-use (RTU) disinfectants in the form of pre-wetted towelettes are increasingly popular among healthcare facilities. Currently, the EPA does not require disinfectant manufacturers to include a recommended maximum surface area per towelette on their product labels. The objective of this study was to investigate the efficacy of disinfectant towelette products on a hard non-porous surface across different coverage areas using a quantitative EPA method. We hypothesized that there would be significant differences in the efficacy of disinfectant towelette products, and that the greater surface area(s) wiped would result in reduced bactericidal efficacy.
This study tested ten disinfectant towelette products against Staphylococcus aureus strain ATCC CRM-6538 and Pseudomonas aeruginosa strain ATCC 15442 on Formica surfaces. Defined surface areas were wiped and the towelette weighed before and after wiping to determine the amount of liquid released. Bactericidal efficacy testing was also performed after wiping following standard EPA protocols.
REFERENCE:
West, Alyssa M et al. “Surface area wiped, product type, and target strain impact bactericidal efficacy of ready-to-use disinfectant Towelettes” Antimicrobial resistance and infection control vol. 7 122. 11 Oct. 2018, doi:10.1186/s13756-018-0416-z

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Synthetic viruses - Anything new?

FRAGMENT: The report of the construction of an infectious horsepox virus from synthesized DNA by Noyce, Lederman, and Evans [1] raised considerable concerns about whether this study will facilitate the construction of smallpox virus (variola) using synthetic biology [2–5]. This is a valid concern, but for a number of reasons—as explained below—no major change concerning the likelihood of a “resurrection” of smallpox emerges from this publication. Having said this, it is also evident that the scientific community, politicians, decision makers, and the lay public have to continue, and probably intensify, a discussion on benefits and risks of synthetic biology in a broader sense.

REFERENCE:
Thiel V. Synthetic viruses-Anything new? PLoS Pathog. 2018 Oct4;
14(10):e1007019. doi: 10.1371/journal.ppat.1007019. eCollection 2018 Oct.

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Biosafety Guidelines

Biosafety guidelines are a set of policies, rules, and procedures necessary to observe by personnel working in various facilities handling microbiological agents such as bacteria, viruses, parasites, fungi, prions, and other related agents and microbiological products. Institutions requiring strict adherence to these biosafety guidelines include clinical and microbiological laboratories, biomedical research facilities, teaching and training laboratories and other healthcare institutions (e.g., clinics, health centers, hospital facilities). These guidelines are intended to provide proper management and regulation of biosafety programs and practices implemented at all levels of the organization. Essential components of the biosafety guidelines contain some or all the following, depending on the facility: microbiological risk assessment and identification; specific biosafety measures, which cover the code of practice, physical plant such as laboratory design and facilities, equipment acquisition and maintenance, medical surveillance, staff training, safe handling of chemicals, with fire, radiation and electricity safety, among others. Additional components may be included such as commissioning and certification guidelines for the facilities. Biosafety guidelines must be made clear, practical and suitable for each facility and must be available for easy reference by all staff, must be reviewed, and updated regularly. While it provides guidance in the application of biosafety practices, this technical guide cannot solely ensure a safe working environment without the commitment of each person to adhere adequately to the biosafety guidelines at all times. Continuous research on biosafety can improve the development of future guidelines[1]
REFERENCE:
Bayot ML, Limaiem F. Biosafety Guidelines. 2019 Jan 31. StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2018 Jan-. PubMed PMID: 30725895.


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Study of Disinfectant Resistance Genes of Pseudomonas aeruginosa

Background: The prevalence of disinfectant resistance in Pseudomonas aeruginosa is on the rise. P. aeruginosa is the most common bacteria isolated from cases of microbial keratitis. Many multi-purpose contact lens disinfectant solutions are available to decontaminate contact lenses before use and to help reduce the incidence of infections. However, with increasing disinfectant resistance, the effect of multi-purpose disinfectant solutions may diminish. The goal of this study was to examine genes associated with disinfectant resistance in ocular isolates of P. aeruginosa and understand the strain’s susceptibility to different multipurpose disinfectant solutions. Methods: Seven potential disinfectant resistance genes were used in BLASTn searches against the whole genomes of 13 eye isolates of P. aeruginosa. A microdilution broth method was used to examine susceptibility to four different multipurpose disinfectant solutions. Results: All strains possessed the sugE2, sugE3 and emrE (qacE) genes. The sugE1 and qacEdelta1 genes were present in 6/13 isolates. No strains contained the qacF or qacG genes. All tested disinfectant solutions had the ability to kill all test strains at 100% concentration, with some strains being susceptible at 1:8 dilutions of the disinfecting solutions. However, the presence of disinfectant resistance genes was not associated with susceptibility to multi-purpose disinfectants. Conclusion: All four tested contact lens disinfectant preparations are effective against P. aeruginosa isolates regardless of the presence of disinfectant resistance genes.
REFERENCE:
Subedi, Dinesh et al. “Study of Disinfectant Resistance Genes in Ocular Isolates of Pseudomonas aeruginosa” Antibiotics (Basel, Switzerland) vol. 7,4 88. 15 Oct. 2018, doi:10.3390/antibiotics7040088


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Protective Gloves and Gowns for Antineoplastic drugs

Background: Many antineoplastic (chemotherapeutic) drugs are known or probable human carcinogens, and many have been shown to be reproductive toxicants in cancer patients. Evidence from occupational exposure studies suggests that health care workers who have long-term, low-level occupational exposure to antineoplastic drugs have an increased risk of adverse reproductive outcomes. It's recommended that, at minimum, nurses who handle or administer such drugs should wear double gloves and a nonabsorbent gown to protect themselves. But it's unclear to what extent nurses do. Purpose: This study assessed glove and gown use by female pregnant and nonpregnant nurses who administer antineoplastic drugs in the United States and Canada. Methods: We used data collected from more than 40,000 nurses participating in the Nurses’ Health Study 3. The use of gloves and gowns and administration of antineoplastic drugs within the past month (among nonpregnant nurses) or within the first 20 weeks of pregnancy (among pregnant nurses) were self-reported via questionnaire. Results: Administration of antineoplastic drugs at any time during their career was reported by 36% of nonpregnant nurses, including 27% who reported administering these drugs within the past month. Seven percent of pregnant nurses reported administering antineoplastic drugs during the first 20 weeks of pregnancy. Twelve percent of nonpregnant nurses and 9% of pregnant nurses indicated that they never wore gloves when administering antineoplastic drugs, and 42% of nonpregnant nurses and 38% of pregnant nurses reported never using a gown. The percentage of nonpregnant nurses who reported not wearing gloves varied by type of administration: 32% of those who administered antineoplastic drugs only as crushed pills never wore gloves, compared with 5% of those who administered such drugs only via infusion. Conclusion: Despite longstanding recommendations for the safe handling of antineoplastic and other hazardous drugs, many nurses—including those who are pregnant—reported not wearing protective gloves and gowns, which are considered the minimum protective equipment when administering such drugs. These findings underscore the need for further education and training to ensure that both employers and nurses understand the risks involved and know which precautionary measures will minimize such exposures.
REFERENCE:
Lawson, Christina C., et al. Antineoplastic Drug Administration by Pregnant and Nonpregnant Nurses. An Exploration of the Use of Protective Gloves and Gowns. AJN The American Journal of Nursing: January 2019 - Volume 119 - Issue 1 - p 28–35

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Sustained Antimicrobial Activity of a Novel Disinfectant Against Healthcare Pathogens

Environmental contamination plays an important role in the transmission of MRSA, VRE, and C. difficile. Suboptimal compliance with hand hygiene or inappropriate glove use can result in indirect transfer of these pathogens to patients. This study evaluates a novel disinfectant that claims to kill microbes on surfaces for ≥24 hours.
REFERENCES:
Rutala, William et al. “1727. Sustained Antimicrobial Activity of a Novel Disinfectant Against Healthcare Pathogens” Open Forum Infectious Diseases vol. 5,Suppl 1 S55. 26 Nov. 2018, doi:10.1093/ofid/ofy209.133

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