Friday, December 31, 2010

Rapid monitoring of bacterial growth and metabolism with the PMEU Unit from clinical, environmental, food and process industry samples



THE PRESENTATION OF PMEU TECHNOLOGY AT THE NEW ZEALAND MICROBIOLOGICAL SOCIETY MEETING 2ND OF DECEMBER 2010, AUCKLAND, WITH THE MOTTO “small things, BIG ideas” BY ADJUNCT PROFESSOR ELIAS HAKALEHTO

ABSTRACT


The idea of constructing a cultivator where microbe enrichment could be started on the sampling site has fascinated us for many years. In the PMEU (Portable Microbe Enrichment Unit) one actually intensifies the research on metabolic reactions in a multiplied scale. This equipment also simulates the interactions between different microbes, and it therefore can be used for studying microbial ecosystems.

Sampling in an Austrian hospital during three years from the bile specimens of about 250 patients indicated that coliformic bacteria and fecal enterococci represented around 60% of the microflora in this part of the gastrointestinal tract. The mixed acid fermenting coliformic strains and the neutral end-products producing Klebsiella/Enterobacter group were shown to live in symbiosis in the PMEU cultures implying to their cooperation in the duodenum. This gives an idea how the microbial community maintains intestinal conditions by regulating the pH and other parameters.

The PMEU Spectrion® equipment validated by the State Research Centre of Finland (VTT) has Ethernet connections which make it possible to follow up the microbe enrichment and detection process in real time. The water department field trials are reported. The PMEU can be used to monitor directly both hygiene indicator strains as well as pathogenic bacteria, both in water and in food. A recent report involving the National Institute of Health and Welfare in Finland describes detection of Campylobacter sp. from natural and household waters using the combination of PMEU enrichment and the real time PCR.

In hospital tests at Kuopio University Hospital in Finland, the PMEU Scentrion® gas sensing bacterial cultivator has been validated for the microbiological blood testing of neonatal and cancer patients. The PMEU has also been used for monitoring the development of the microflora after the birth or antibiosis. The enhanced enrichment could be used effectively for improving the recovery of bacterial cells in the detection process.

Wednesday, December 22, 2010

Biocide testing (KILL tests) with PMEU Spectrion


PMEU Spectrion ® can be applied as a rapid tool for two types of biocide testing.


GROWTH PREVENTION TEST shows the ability of the biocides to prevent microbial growth in processes. Short detection times (typically under 10 hours) indicate poor or lacking effect of the active compound. Long detection times (over 10 hours) show that the biocide can prevent the microbial growth in the target.

This test can be performed in aerobic, microaerophilic or anaerobic conditions.

Temperature control of PMEU unit allows testing of psycrophilic, mesophilic and thermophilic microorganisms.

Turbidity of the test syringes, caused by microbial growth, will be documented automatically and growth curves will be drawn for every syringe. Detection time
(= the moment of the test, counted from the beginning, when the turbidity has increased by a certain factor) can be seen from these growth curves.

PMEU KILL TEST bases on the biocide-dependent change of microbial count. ATP Assay is the fastest method to detect the quantitative increase and the decrease of microbial cells in the samples and is therefore chosen to assist PMEU biocide testing in cases where both the growth prevention and the death of microbes shall be evaluated.

Results of these two types of biocide tests have a significant correlation.

GROWTH PREVENTION TEST can visualize some microbial processes which cannot be detected with traditional colony count method, eg. it shows the prevention of spore germination to vegetative cells by an effective biocide.

All PMEU Spectrion® methods have one excellent property: they indicate the effects of biocides in the original samples, and recovering effect of colony count methods are totally excluded. The user can focus the attention to the biocidic effects which really take place in the paper machine processes.

Modifications of PMEU biocide testing for biofilm control are also available.

Saturday, November 20, 2010

Demonstration of PMEU Spectrion coliform analyses in Tornio, 17th - 18th November 2010.




Finnish Water and Waste Water Works Association (FIWA) has arranged a conference for the professionals of water and wastewater authorities and salesmen, especially those locating in Northern Finland, in Tornio at the end of November 2010. FIWA's membership includes over 300 Finnish water utilities which cover about 90 % of water services in Finland. In addition FIWA has about 130 collaborating members (companies, institutes, etc.), among them also SAMPLION Ltd.

Rapid, quantitative analyses of coliforms and E.coli, based on COLILERT cultivation (IDEXX, U.S.A.) were demonstrated on the expo stand of SAMPLION Ltd. These methods, also applied to the national POLARIS Project in Finland by SAMPLION Ltd. and BERNER Ltd., have been visualized in the photos above. Yellow color of the cultures indicate coliform growth and E.coli can be detected among other coliforms by its blue fluorescence when illuminated with short-wave UV emission ("Wood's Lamp).

A short, oral presentation concerning the benefits of broth culture analyses (compared to colony count analyses) was held by Key Account Director Juha V.Mentu who also highlights those PMEU Spectrion features which are unique among broth culture methods: speed (which has been achieved with efficient mixing and aeration), alternative gas compositions (for aerobic, microaerophilic and anaerobic microorganisms) and ON LINE growth measuremets from the test syringes, deliverable also in wireless way to the observer of the analyses.

Thursday, June 24, 2010

Hygiene Monitoring with the Portable Microbe Enrichment Unit (PMEU)

Elias Hakalehto, Doc., Ph.D.
Department of Biosciences
University of Eastern Finland
P.O.B. 1627
FI-70211 Kuopio
Finland

Abstract, 41st R3-Symposium of the R3-Nordic Society in Espoo 25.-26.5.2010

The PMEU (Portable Microbe Enrichment Unit) technology (Samplion Oy, Siilinjärvi, Finland) is a microbe cultivation method producing advantageous growth conditions for individual bacterial cells. Therefore, monitoring of bacterial populations and the presence of potential contaminants is optimised in terms of speed and accuracy. Hygienically important isolates of various bacterial strains have been cultivated aerobically, microaerobically or anaerobically using standard broth media by standard PMEU enhanced enrichment technology, or by PMEU Spectrion ® or PMEU Scentrion ® sensored units equipped with optical, IR, or gas sensors. The units were produced by Samplion Oy according to the ISO 9001 accepted protocol. The reference cultivations were carried out using standard microbiological procedures. The PMEU Spectrion ® is being validated by the VTT of Finland. PMEU versions facilitated ultra-fast detection of coliformic bacteria, bacilli, salmonellas, staphylococci, streptococci, campylobacteria and other groups. The PMEU Scentrion ® equipped with gas sensors for volatile organic compounds detected the contaminants at concentrations of bacterial levels around 10-1000 cfu/ml in 2-5 hours. Also the hospital validation studies in Austria and Finland are summarized. Moreover, The PMEU has been demonstrated to help the environmentally stressed cells to recover and become viable in the enrichment cultures. Hygiene sampling was carried out using a specific sampling syringe serving also as an incubator in the PMEU.

Tuesday, June 15, 2010

Biocide testing with PMEU Spectrion has begun!



PMEU biocide testing has been available already since the early years of this millenium. This method is based on Basic PMEU and ATP Assay. This combination gives reliable results and has been presented in several articles (last time in Paper and Timber, Nov. 2009).

First, very promising tests with PMEU Spectrion have now started, both in laboratory and in the mills. The picture above shows a typical equipment for mill trials, containing both PMEU Spectrion and ATP Assay tests. PMEU Spectrion measures growth curves of the microbial population (total or selective) automatically and is therefore independent of ATP Assay.

SAMPLION Ltd. will focus on Pulp and Paper biocide tests in August 2010, and samples are very welcome from 2.8.2010 on. More information of this free chance to be familiar with this novel method will be delivered by Microbiologist, Key Account Manager Juha V. Mentu (juha.mentu@samplion.fi) by request.

Friday, June 4, 2010

SAMPLION Ltd. exhibition stand before opening of the congress.


This picture represents the stand of SAMPLION Ltd. in PulPaper 2010 Congress exhibition on the day before the opening. This event was a success! 5800 visitors on Wednesday, on the "middle day" of the congress!

In addition to the exhibition stand, an oral presentation was given by Key Account Manager (originally paper industry microbiologist) Juha V. Mentu, which will be sent by request.

Thursday, May 27, 2010

Environmental Health Conference in Kuopio, Finland, 27.-28.5.2010.



PMEU Spectrion interested the participants of Environmental Health Conference in Kuopio, Finland. Visitors of exhibition were able to see the selective cultivation of E.coli which can be confirmed by its blue fluorescence when illuminated with "Wood's lamp", emitting UV to the target syringe.

Laboratory Manager Anneli Heitto (left) and Laboratory Engineer Maarit Miskala, both working for FINNOFLAG Ltd, were representing PMEU method in the exhibition. Maarit Miskala also had an oral presentation about the control of swimming waters where PMEU Spectrion significantly helps to collect analytical data faster and with much higher sampling frequence than the traditional laboratory ananlyses.

Monday, May 24, 2010

New SAMPLION website in English is open to visit.



SAMPLION website has now been translated to English. You are very welcome to visit

www.samplion.fi

where you can find further information about Advantages to users, Results, Equipment, Application areas as well as to find our personnel to contact.

Welcome!

Sunday, May 16, 2010

SAMPLION Ltd. joins the international PulPaper Exhibition and Conference at the Helsinki Exhibition & Convention Centre 1.-3.6.2010



PMEU methods have been applied in paper industry microbiology since 2002 by company's microbiologists. The main facts and procedures of PMEU biocide and biofilm testings have been presented in Paper and Timber, Vol. 91 / No. 7-8 / 2009.

SAMPLION Ltd. will join PulPaper 2010, 1.-3.6.2010, Helsinki as an exhibitor and welcomes You to visit the company's exhibition stand,

6b27


The reference from the webpages of PulPaper tells "The event, organized by Adforum in cooperation with the Paper Engineers’ Association, AEL and The Finnish Fair Corporation will be the largest and most important show for the pulp, paper, board and converting industry worldwide in 2010. Exhibitors and visitors travel from all over the world to take part in this international event. The last PulPaper show in 2007 attracted 16102 visitors from 78 countries!".

Sunday, April 25, 2010

A new era in microbiology: back to Pasteur's times!

The history of microbiology contains several eras with different targets. I will refer them in this way (based on my over 30 years experience as microbiologist and teacher of microbiology and biochemistry):

In the beginning, cultivation and observation of microorganisms was the main target. Doctors like Pasteur and Koch were very innovative and developed intelligent culture medias and vessels to perform very delicate experiments. The everlasting fight against pathogenic microbes was the primary target but Louis Pasteur started to help eg. wine producers to solve their quality problems, caused by microbes.

The combination of microbiology and biochemistry on the second era was very satisfying by solving questions concerning the huge amount of anabolic and catabolic processes included in microbial growth. More and more were also learned in the area of microbial ecology. Questions like "who? what? when? where? how? why?" were partially solved (ref. MADSEN,E.L. 2008. Environmental Microbiology. From Genomes to biochemistry. Blackwell Publishing).

"Third era" can be described by the novel methods to identify bacteria. Biochemical test kits (API etc.) were replaced by Fatty Acide Methylated Ester method (FAME) by Hewlett-Packard on 80's. After it, molecular biology methods, based on ribosomal RNA and DNA, helped to construct the development lines of microorganisms.

Today is the time of new era. We know the "family trees" of bacteria but we should now continue with environmental microbiology and microbial ecology to solve questions like "How, why, by whom and in which conditions will the raw materials of paper industry be biodeteriorated?", "How can we prevent these processes by setting the process conditions unsuitable for those biochemical processes?", "Can we prevent the growth of biofilms and slimes in an ecological way?", "How to prevent selectively the growth of toxin producers like Bacillus cereus in paper and board processes?", how to fight against Legionella in paper industry?".

Names are not the most important thing. Most important is, how the bacteria act in different ecological niches of a paper machine. This work has to be done by using simulations of paper processes which is possible by wet end simulators of research units (as an example: VTT in Jyväskylä, Finland) and laboratory/field instruments (like biofilm detectors in the processes or PMEU incubators by Samplion Ltd.).

The role of PMEU is getting more and more important because this method helps to detect microbial growth of different types (biofilms included) in a very short period of time as well as to test simultaneously the effects of alternative biocides in small-scale tests whose growth parameters match with the growth conditions in the real processes.

We are - and we shall - turn back to the era of Pasteur & Koch: the names are already known, and we shall now investigate, what the contaminating microbes are doing in the industrial processes and how to prevent losses of raw material, machine stops and poor quality of the products by simulating growth processes in small-scale tests, performed in the laboratory or in the field, by the machies themselves.

Saturday, March 27, 2010

PMEU applications for water hygiene analyses have been published in "Rakennustarkastuspäivät 2010"



Novel PMEU water hygiene analyses, intended in the quantitative control of total coliforms, E.coli and enterococci, have been published in "Rakennustarkastuspäivät 2010" Conference, Kuopio, Finland at the end of March. This annual event is dedicated to professionals of building regulations and related administration and collected over 200 participants to the beutiful capital of Savo region, in the middle of Saimaa lake district.

SAMPLION Ltd. was exhibiting PMEU Spectrion and its water hygiene applications, now qualified by VTT, alongside Suomen Jätevesi Ltd. which is the major supplier of small-scale waste water purification systems in Finland.

The picture above demonstrates the combination of selective growth medium and MUG-based UV fluorescence confirmation of E.coli. Among the three PMEU syringes, only the one on the left, containing a culture of E.coli, indicates this fluorescence. This PMEU method is now approved by VTT, Technical Research Centre of Finland (VTT-S-01705-10) and it also follows the guidelines of international standardization (SFS-EN ISO 9308-3) to fulfill the needs of rapid, quantitative control of hygiene indicator bacteria.

Wednesday, March 3, 2010

Detection Time as the Variable in Quantitative Microbe Analyses



There are several methods to quantify the count of microbes. Historically the very first of them (if microscopy is excluded) was the use of dilutions series in nutrient broths. This methods was later developed to MPN (Most Probable Number) Method, which was the standard until the rise of Colony Count methods. Colony counts give accurate numbers but suffer of the poorer recovery of the microbial cells, compared to the broth methods. It is also unavailable for certain, difficult types of samples.

"Third Generation" of quantitative mb analyses was established some 30 years ago. They are based on analytical methods which give the results much faster than the traditional methods by measuring some chemical or physical features of the samples which are in a relationship with the count (or activity) of microbes of the original samples.

The increase of the nutrient broth turbidity, caused by the growth of microorganisms, is maybe the most common "3rd Generation" method used today. It is an optical method which gives levels of either total growth or specified microbes (in latter case, selective broths with confirmation tests are applied). An inversed corretation between microbial load and detection time (= the moment of the testing period when turbidity begins to rise) can be found and correlation lines/tables between colony counts and detection times are easy to construct.

The figure above is an example of the growth curves, derived by PMEU Spectrion. X axis shows the time scale of the test, y axis indicates the values of RT. This curve shows that PMEU Spectrion actually illustrates the classic growth curves with the lag, log and stat phases (and, if the cultivation will continue long enough, also the kill phase). The moment when the turbidity begins to rise is inversely related to the count of microbes in the original sample.

Tuesday, January 12, 2010

SAMPLION Ltd. in Finnish Medicine Congress, Helsinki, January 11.-14.2010



PMEU Spectrion is one of the microbiological analyse devices manufactured by SAMPLION Ltd. It represents a modification of the basic PMEU where the turbidity of the samples, correlating with the microbial growth inside the test syringes, will be automatically measured with LED detection.

Beside the PMEU Spectrion, results of some hospital projects were also presented in the form of a poster.

Sunday, January 3, 2010

Use of portable enrichment unit in rapid characterization of infantile intestinal enterobacterial microbiota

Jouni Pesola‌1,2, Outi Vaarala‌3, Anneli Heitto‌4 & Elias Hakalehto‌4,5
Institute of Clinical Medicine, Pediatrics, University of Kuopio, Kuopio
Department of Pediatrics, Kuopio University Hospital, Kuopio
National Institute for Health and Welfare, Helsinki
Department of Biosciences, University of Kuopio, Kuopio
Finnoflag Oy, Kuopio and Siilinjärvi, Finland
Correspondence: Jouni Pesola, Department of Pediatrics, Kuopio University Hospital, POB 1777, FI-70211 Kuopio, Finland. E-mail: jouni.pesola@kuh.fi


Abstract

Objective: The aim of this study was to develop a practical and efficient method for isolation and characterization of the enterobacterial microbiota of infant fecal samples. It could be used for reliable comparisons between individual microflora in different neonates, and in the follow-up of the succession in the floral developments in various nutritional conditions. Methods: A combination of traditional plate culture and different enrichment procedures carried out in a portable microbe enrichment unit (PMEU) was studied. The strains were isolated by plate culture both before and after enrichment. The isolated strains were identified by biochemical tests and the phenotypes of the isolates were compared by the PhenePlate™ technique. To obtain a picture of the formation of intestinal flora during the first year two follow-up samples were collected at the ages of 3 and 12 months from eight Finnish infants (one girl and seven boys). Results: Altogether 21 and 46 enterobacterial isolates were detected in samples taken at the ages of 3 and 12 months, respectively; 7/21 (33%) and 24/46 (52%) isolates were not detected by direct plating but only after enrichment. This material was supposed to demonstrate the differences between the different methodological approaches. Conclusion: There were age-dependent changes in the species proportions of different subpopulations of Enterobacteriaceae detected from the infantile fecal microbiota at different times. The statistically significant difference in the number of isolations with and without enrichment step suggests that the use of enrichment is highly recommendable and the PMEU equipment and method have now proven suitable for that purpose, at least for screening and monitoring the infant fecal flora. The method offers more representative, quicker, and relatively practical tools for obtaining an overall picture of the enterobacterial microbiota of the fecal samples and its variations, thus also giving an idea of the microbial communities in the gastrointestinal tract.