Use of ultrasonic energy in assessing microbial contamination on surfaces

Ultrasonic tanks were evaluated for their ability to remove viable microorganisms from various surfaces for subsequent enumeration. Test surfaces were polished stainless steel, smooth glass, frosted glass, and electronic components. The position of contaminated surfaces in relation to the ultrasonic energy source, distance of the ultrasonic source from the test surfaces, and temperature of the rinse fluid were some of the factors which influenced recovery. Experimental systems included both naturally occurring microbial contamination and artificial contamination with spores of Bacillus subtilis var. niger. The results showed that ultrasonic energy was more reliable and efficient than mechanical agitation for recovering surface contaminants. Conditions which increased the number and percentage of microorganisms recovered by ultrasonic energy were: using a cold rinse fluid, placing the sample bottle on the bottom of the ultrasonic tank, and facing the contaminated surfaces toward the energy source. It was also demonstrated that ultrasonic energy could be effectively used for eluting microorganisms from cotton swabs.     

Kitchen hygiene in daily life

More and more data are available indicating that numerous (infectious) diseases are related to the home environment. Airborne microorganisms (bacteria, fungi), mites (in sheets or carpets), and (parts of) insects and beetles may be the cause of respiratory diseases such as asthma. In Europe and North America, more than half of the registered food infections appear to be contracted in the home. Then there is the development of new pathogens or the adaptation of microorganisms to extreme conditions. In many cases, the occurrence of adapted or resistant microorganisms is a reaction to the changing environment. Examples of this are the growth of Listeria monocytogenes in refrigerated food products and the presence of Legionella pneumophila in systems containing stagnant water.The main sources of infection in the domestic environment are people, pests, pets, and contaminated food and water. Germs are transmitted by direct contact with people or animals, by contaminated food, water, surfaces and air.Under circumstances favourable to microorganisms, the cells are able to survive or multiply into large numbers. Especially in places which stay moist for a long time considerable amounts of microorganisms are found, among with pathogenic types.The phrase ‘home hygiene’ does not merely refer to cleaning the house (daily). In practice, cleaning is not the only important issue; knowing how to prevent contamination is just as crucial. Domestic hygiene is the total sum of the measures used to prevent (insofar as possible) contamination with pathogens, and thus decreasing the number of infectious diseases. The hygiene measures required can be divided into three groups:

• Hygiene during food preparation.

• Personal and sanitary hygiene.

• Domestic environment.

This contribution will focus on food storage, food preparation and the effect of 'hygienic cleaners’ in the domestic kitchen.     

Perspectives on air disinfection methods in the workplace of blood transfusion services

Blood collection and preparation is a relatively open operation in a conventional environment, and is vulnerable to be contaminated by various types of airborne pathogenic microorganisms. It is important to establish stable and effective air disinfection methods for all types of environments in blood transfusion services, in order to control air hygiene quality and thus reduce the probability of contamination during blood collection. This paper analyzes and summarizes the principles, advantages, and disadvantages of commonly used chemical and physical air disinfection methods and their application status. It is suggested that over-reliance on chemical reagents and disinfection facilities be reduced, so that better results can be achieved with the combination of multiple disinfection methods and dynamic air hygiene monitoring.     

Microbial Stability of Pharmaceutical and Cosmetic Products

This review gives a brief overview about microbial contamination in pharmaceutical products. We discuss the distribution and potential sources of microorganisms in different areas, ranging from manufacturing sites, pharmacy stores, hospitals, to the post-market phase. We also discuss the factors that affect microbial contamination in popular dosage forms (e.g., tablets, sterile products, cosmetics). When these products are contaminated, the microorganisms can cause changes. The effects range from mild changes (e.g., discoloration, texture alteration) to severe effects (e.g., changes in activities, toxicity). The most common method for countering microbial contamination is the use of preservatives. We review some frequently used preservatives, and we describe the mechanisms by which microorganisms develop resistance to these preservatives. Finally, because preservatives are inherently toxic, we review the efforts of researchers to utilize water activity and other non-preservative approaches to combat microbial contamination.     

Microbial contamination of BM products before and after processing: a report of incidence and immediate adverse events in 257 grafts

BACKGROUND: The incidence and potential clinical consequences of bacterial contamination of autologous and allogeneic BM products remains open to question. We report our experience of bacterial contamination of BM grafts and adverse events that occurred after transplantation. METHODS: From January 2003 to February 2006, 257 BM harvests were processed and infused at our institution. Analysis of microbial contamination incidence before and after processing, sensitivity spectra of isolated bacteria and adverse events after graft infusion were analyzed. RESULTS: Nineteen of 257 BM (7.4%) were contaminated. Coagulase-negative Staphylococcus (n=9) and Propionibacterium acnes (n=6) were the most frequently isolated microorganisms. Two of nine coagulase-negative staphylococci were found to be resistant to erythromycin and two of six P. acnes to fosfomycin and gentamycin. The frequency and severity of immediate adverse events reported in patients receiving a contaminated graft were similar to those observed in patients receiving a non-contaminated product. No major adverse sequelae occurred after infusion of contaminated grafts. Finally, none of the patients transplanted with a contaminated graft developed bacteriemia that could have been related to the isolated microorganism. DISCUSSION: Microbial contamination of BM progenitor cell grafts does not induce severe clinical complications or infectious diseases after infusion. The vast majority of isolated pathogens were skin contaminants.     

Forensic application of microbiological culture analysis to identify mail intentionally contaminated with Bacillus anthracis spores

The discovery of a letter intentionally filled with dried Bacillus anthracis spores in the office of a United States senator prompted the collection and quarantine of all mail in congressional buildings. This mail was subsequently searched for additional intentionally contaminated letters. A microbiological sampling strategy was used to locate heavy contamination within the 642 separate plastic bags containing the mail. Swab sampling identified 20 bags for manual and visual examination. Air sampling within the 20 bags indicated that one bag was orders of magnitude more contaminated than all the others. This bag contained a letter addressed to Senator Patrick Leahy that had been loaded with dried B. anthracis spores. Microbiological sampling of compartmentalized batches of mail proved to be efficient and relatively safe. Efficiency was increased by inoculating culture media in the hot zone rather than transferring swab samples to a laboratory for inoculation. All mail sampling was complete within 4 days with minimal contamination of the sampling environment or personnel. However, physically handling the intentionally contaminated letter proved to be exceptionally hazardous, as did sorting of cross-contaminated mail, which resulted in generation of hazardous aerosol and extensive contamination of protective clothing. Nearly 8 x 10(6) CFU was removed from the most highly cross-contaminated piece of mail found. Tracking data indicated that this and other heavily contaminated envelopes had been processed through the same mail sorting equipment as, and within 1 s of, two intentionally contaminated letters.     

Microbiological Laboratory Hazard of Bearded Men

An investigation was conducted to evaluate the hypothesis that a bearded man subjects his family and friends to risk of infection if his beard is contaminated by infectious microorganisms while he is working in a microbiological laboratory. Bearded and unbearded men were tested with Serratia marcescens and Bacillus subtilis var. niger. Contact aerosol transmission from a contaminated beard on a mannequin to a suitable host was evaluated with both Newcastle disease virus and Clostridium botulinum toxin, type A. The experiments showed that beards retained microorganisms and toxin despite washing with soap and water. Although washing reduced the amount of virus or toxin, a sufficient amount remained to produce disease upon contact with a suitable host.     

The Ethological–Physiological Response of <Emphasis Type="Italic">Apis mellifera</Emphasis> L. Bees to Carbohydrate Lead Contamination

The response of bees to lead contamination of a carbohydrate feed has been studied. It was established that Pb in sucrose solution had a toxic and repelling effect on the bees. It was found that the amount of contaminated feed consumed by the bees decreased with a corresponding increase in the content of Pb in it. It is noted that this element accumulates unevenly in different body parts and parts of the digestive tract of the bees, thereby affecting their metabolic activity and accelerating processes of physiological aging, and the specific reaction of the bees to contaminated feed is expressed in their rejection of its consumption and in inhibition of their locomotion.     

Forensic Application of Microbiological Culture Analysis To Identify Mail Intentionally Contaminated with Bacillus anthracis Spores

The discovery of a letter intentionally filled with dried Bacillus anthracis spores in the office of a United States senator prompted the collection and quarantine of all mail in congressional buildings. This mail was subsequently searched for additional intentionally contaminated letters. A microbiological sampling strategy was used to locate heavy contamination within the 642 separate plastic bags containing the mail. Swab sampling identified 20 bags for manual and visual examination. Air sampling within the 20 bags indicated that one bag was orders of magnitude more contaminated than all the others. This bag contained a letter addressed to Senator Patrick Leahy that had been loaded with dried B. anthracis spores. Microbiological sampling of compartmentalized batches of mail proved to be efficient and relatively safe. Efficiency was increased by inoculating culture media in the hot zone rather than transferring swab samples to a laboratory for inoculation. All mail sampling was complete within 4 days with minimal contamination of the sampling environment or personnel. However, physically handling the intentionally contaminated letter proved to be exceptionally hazardous, as did sorting of cross-contaminated mail, which resulted in generation of hazardous aerosol and extensive contamination of protective clothing. Nearly 8 × 10⁶ CFU was removed from the most highly cross-contaminated piece of mail found. Tracking data indicated that this and other heavily contaminated envelopes had been processed through the same mail sorting equipment as, and within 1 s of, two intentionally contaminated letters.     

Despersion and survival ofLegionella in air

Summary From the time of the report of the Philadelphia outbreak in 1976, the airborne contamination of patients through cooling towers had been suspected although never truly corroborated. Since then, contamination through different air conditioning systems or cooling towers has often been implicated in both epidemics and endemics as well as in sporadic cases. Whatever the conditions, contaminated water present in the different air conditioning systems remains the main source of infection. From the contaminated sites an aerosol occurs. The infectivity of the aerosol is linked to the size of the droplets (d≥μm). A close surveillance is mandatory and preventive measures have to be tanken.     

An in-use study of the relationship between bacterial contamination of food preparation surfaces and cleaning cloths

The relationship between bacterial contamination of food preparation surfaces and the cleaning cloths associated with these surfaces was investigated in a busy college kitchen. Cleaning cloths used in conjunction with detergent became heavily contaminated within hours of first use. Following 'cleaning' with these cloths with detergent, both surfaces and cloths were found to be more heavily contaminated and there was evidence of contamination transfer to and from cloths and surfaces. Following 'cleaning' with cloths impregnated with quaternary ammonium disinfectant, there was a significant reduction in contamination on both surfaces and cloths, together with a reduction in the numbers of enterobacteria and pseudomonads. The results suggest that significant improvements in kitchen hygiene could be achieved by ensuring that contaminated cloths are not used for food preparation activities.     

Method and equipment for detection of pathogenic microorganisms using tracking analytical membranes

Method for detection of pathogenic microorganisms in different microbiological assays (water, faeces, punctate etc.) has been developed. Device for preparation of samples and filtration of studied material through transparent tracking membranes with fixed size of pores and subsequent microscopy of filtrate was developed also as well as device for filtration of different liquids in closed cylinder under pressure. Method significantly increases informativeness and reliability of analyses, and decreases of their labor and financial costs.     

RDX degrading microbial communities and the prediction of microorganisms responsible for RDX bioremediation

The explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) has caused significant soil and groundwater contamination. To remediate these sites, there is a need to determine which microorganisms are responsible for in situ biodegradation of RDX to enable the appropriate planning of bioremediation efforts. Here, studies are examined that have reported on the microbial communities linked with RDX biodegradation. Dominant microorganisms across samples are discussed and summarized. This information is then compared to current knowledge on RDX degrading isolates to predict which organisms may be responsible for RDX degradation in soils and groundwater. From the phyla with known RDX degrading isolates, Firmicutes and Proteobacteria (particularly Gammaproteobacteria) were the most dominant organisms in many contaminated site derived samples. Organisms in the phyla Deltaproteobacteria, Alphaproteobacteria and Actinobacteria were dominant in these studies less frequently. Notably, organisms within the class Betaproteobacteria were dominant in many samples and yet this class does not appear to contain any known RDX degraders. This analysis is valuable for the future development of molecular techniques to track the occurrence and abundance of RDX degraders at contaminated sites.     

The effect of foliar application of urea on the root fungi of wheat growing in soil artificially contaminated withFusarium spp.

Repeated treatment of overground parts of wheat with a solution of urea was reflected in an increased count of bacteria and a decreased count of fungi on the roots of plants grown in soil contaminated with the fungusFusarium spp., which is known to be noxious to plants. The occurrence of some fungal genera on the roots of plants growing in contaminated and uncontaminated soil after foliar application of urea was also changed. The roots of treated planted were found to contain more bacteria with a lytic effect and retarding the growth of the fungus used for the contamination. The favourable effect of foliar application of urea on wheat was reflected in an increased dry weight of plants even if growing in a soil contaminated withFusarium.     

Residual Antibiotics in Decontaminated Human Cardiovascular Tissues Intended for Transplantation and Risk of Falsely Negative Microbiological Analyses

We investigated the presence of antibiotics in cryopreserved cardiovascular tissues and cryopreservation media, after tissue decontamination with antibiotic cocktails, and the impact of antibiotic residues on standard tissue bank microbiological analyses. Sixteen cardiovascular tissues were decontaminated with bank-prepared cocktails and cryopreserved by two different tissue banks according to their standard operating procedures. Before and after decontamination, samples underwent microbiological analysis by standard tissue bank methods. Cryopreserved samples were tested again with and without the removal of antibiotic residues using a RESEP tube, after thawing. Presence of antibiotics in tissue homogenates and processing liquids was determined by a modified agar diffusion test. All cryopreserved tissue homogenates and cryopreservation media induced important inhibition zones on both Staphylococcus aureus- and Pseudomonas aeruginosa-seeded plates, immediately after thawing and at the end of the sterility test. The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media. Based on standard tissue bank analysis, 50% of tissues were found positive for bacteria and/or fungi, before decontamination and 2 out of 16 tested samples (13%) still contained microorganisms after decontamination. After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method. When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated. Antibiotic residues present in tissue allografts and processing liquids after decontamination may mask microbial contamination during microbiological analysis performed with standard tissue bank methods, thus resulting in false negatives.     

Correlation of in vitro challenge testing with consumer use testing for cosmetic products

An in vitro microbial challenge test has been developed to predict the likelihood of consumer contamination of cosmetic products. The challenge test involved inoculating product at four concentrations (30, 50, 70, and 100%) with microorganisms known to contaminate cosmetics. Elimination of these microorganisms at each concentration was followed over a 28-day period. The test was used to classify products as poorly preserved, marginally preserved, or well preserved. Consumer use testing was then used to determine whether the test predicted the risk of actual consumer contamination. Products classified by the challenge test as poorly preserved returned 46 to 90% contaminated after use. Products classified by the challenge test as well preserved returned with no contamination. Marginally preserved products returned with 0 to 21% of the used units contaminated. As a result, the challenge test described can be accurately used to predict the risk of consumer contamination of cosmetic products.     

Ecological risk assessment of metals in roadside agricultural soils: A modified approach

The threat of metal contamination of roadside agricultural soils poses severe ecological risks throughout the globe due to dual contamination by intensive agriculture and traffic-related activities. Therefore, the present study was conducted to assess ecological risks posed by metals (Cd, Co, Cr, Cu, Fe, Mn, Pb, and Zn) in intensively cultivated roadside agricultural soils of Punjab, India. The results of the contamination assessment factors i.e. Contamination Factor (CF) and Enrichment Factor (EF) indicated that the studied soils were moderately to highly contaminated with metals. It was observed that the Modified Risk Index (MRI), which is based upon EF, indicated the ecological risks of metals more efficiently than the traditional Ecological Risk Index (RI), which is based upon CF in the present study. The MRI values indicated that the metals posed considerable to high risk in 67.86% samples as compared to only 7.14% samples based upon RI values. Thus, MRI can be considered as a more efficient risk indicator in comparison to RI. Spatial distribution maps of MRI indicated to higher metal contamination in inner urban parts of study area due to higher traffic and industrial activities.     

一种简便、高效、经济的从凝胶中回收DNA的方法

目的:尝试一种简便、高效的从凝胶中回收DNA的方法。方法:在Eppendorf管的管底用注射器扎孔,将一小团用Eppendorf管融化后拉成的细丝放入管中,把含有DNA的凝胶放在细丝上,离心,收集从管底流出的液体,经酚氯仿抽提后用乙醇沉淀DNA。结果:经过简单的离心即可近乎100%地回收凝胶中的DNA。结论:使用该方法从琼脂糖凝胶回收DNA,操作简单,回收率高,无其他试剂污染。     

Microbiological contamination in genetically modified animals and proposals for a microbiological test standard for national universities in Japan.

The Biosafety Committee of the Japanese Association of Laboratory Animal Facilities of National Universities (JALAN) investigated recent episodes of microbiological contamination in genetically modified mice (GMM), and the countermeasures taken when the contaminated GMM were introduced into animal facilities, by questionnaires addressed to 53 animal facilities belonging to JALAN and serological tests. Although almost all of the contaminated GMM were accepted with conditions such as rederivation after or before reception and housing in designated rooms, contamination with a spectrum of microorganisms was demonstrated in GMM transferred domestically and from abroad. In serological tests, Mycoplasma pulmonis, mouse parvovirus, and mouse encephalomylitis virus were detected in GMM transferred from domestic facilities and from abroad. The present results of the questionnaires and serological tests suggest that GMM are highly and widely contaminated with microorganisms compared with mice from commercial breeders. Thus, we propose a microbiological requirement, including microbiological status--excellent, common, and minimum--as a guide for the transfer and procurement of mice and rats in Japan.     

Feasibility of the laryngeal tube airway for artificial ventilation in pigs and comparison with the laryngeal mask airway

Airway management in anesthetized pigs is known to be technically demanding, and the 'gold standard' technique of endotracheal intubation is particularly difficult to master. The authors investigated the feasibility of the laryngeal tube as an alternative technique for airway management in German Landrace pigs (n = 5). They compared this method with the laryngeal mask, which is considered to be an effective yet relatively straightforward tool for porcine airway management. One after the other, investigators attempted to establish an airway in each anesthetized, artificially ventilated pig using each device. The laryngeal tube was too short to intubate the largest pig (weighing 45 kg), and it took investigators slightly longer to insert this device compared with the laryngeal mask. With the laryngeal mask, there were several incidents of gastric insufflation. Despite these complications, all investigators were able to establish a secure airway and maintain oxygenation with the laryngeal tube, and all subjectively rated both devices as easy to use.