Friday, July 17, 2009

Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR

Tarja Pitkänen, Juliane Bräcker, Ilkka T. Miettinen, Anneli Heitto, Jouni Pesola, and Elias Hakalehto

Can. J. Microbiol. 55(7): 849–858 (2009):

Abstract: An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

Key words: Campylobacter, enrichment, Portable Microbe Enrichment Unit, real-time PCR, water.

Tuesday, July 14, 2009

PMEU-teknologian perusteet / Basic descriptions of PMEU technology.

PMEU -teknologia on tarkoitettu nopeuttamaan merkittävästi standardien mukaisia mikrobien viljelymenetelmiä. Samalla mikrobit, kuten erilaiset bakteerit, homeet ja hiivat, saadaan nopeasti rikastettua molekyylibiologisten tutkimusmenetelmien vaatimiin pitoisuuksiin. PMEU-menetelmää onkin menestyksellisesti hyödynnetty mm. PCR:n,immunoanalyysien ja ATP -mittausten esikäsittelymenetelmänä.

PMEU:n avulla saadaan mikrobien tutkimuksesta

1. herkempää
2. nopeampaa
3. luotettavampaa

kuin ennen; laboratorioissa, klinikoilla, tuotantolinjastoilla ja maastossa. Sen avulla näytteet saadaan tutkimuksen kohteiksi heti, ilman näytteen laatua heikentäviä viiveitä. Myös ympäristöstressistä kärsineet solut saadaan tehokkaasti elpymään.

FT, MMM Elias Hakalehto
Mikrobiologi, bioteknologi
International Postgraduate Diploma in Biotechnology (U.K.)
Bioteknisen mikrobianalytiikan dosentti
Kuopion yliopisto



PMEU (Portable Microbe Enrichment Unit) technology has been designed tomake the standard microbe cultivation methods significantly faster. At the same time the microbes, such as bacteria, moulds and yeasts, are rapidly enriched to concentrations required for different molecular methods. PMEU method has been successfully used for pretreatments for the PCR,
immunoanalysis and ATP measurements.

PMEU approach makes the microbiological research

1. more sensitive
2. faster
3. more reliable

than before; in the laboratories, clinics, production lines and outdoors.
The samples are studies immediately after sampling without delays that
lower the sample quality. Also the cells suffered from environmental
stresses are effectively recovered.

Elias Hakalehto, PhD, MSc (Agr.Sci)
Microbiologist, Biotechnologist
International Postgraduate Diploma in Biotechnology (U.K.)
Docent in Biotechnical Microbe Analytics
University of Kuopio
Kuopio, Finland

Monday, July 13, 2009

PMEU applications for paper industry microbiology.

Paper industry microbiology is one of the several sectors of microbiology which can have significant benefits when applying PMEU methods for rapid hygiene control and HACCP procedures.

A paper mill consists of several technical parts which all act as growth environments (niches) for microbes. Paper machine itself is like a continuous flow fermentor, securing constant nutrient supply, growth environment and feedback for microbes. All regions of the system have their own microbial populations which cause risks for the processes, machinery, employees and product hygiene. Portable Microbiological Enrichment Unit (PMEU) by SAMPLION Oy (Siilinjärvi, Finland) has been modified to an analytical tool for paper industry microbiology, intended in estimation of microbial activities in process waters and simultaneous evaluation of alternative paper machine biocides. ATP Assay was combined with PMEU incubations to give quantitative values of living biomass during tests. PMEU can be used for the detection of harmful bacteria in raw materials and processes as well as on the control of biofilm-forming microbes when epifluorescence microscopy for the observations of biofilm growth on test plates is available. PMEU method helps to avoid bias, caused by the selecting and recovering effects of traditional colony count methods. Because of the robustness of  PMEU incubator, it does not necessarily need any specified microbiological laboratory but acts also as a stand alone cultivation system in common paper mill laboratories research institutes as their additional microbiological tool.

Sunday, July 12, 2009

Marketing of PMEU unit is starting in this summer.

PMEU (Portable Microbiological Enrichment Unit) was developed by Dr. Elias Hakalehto for research purposes on 90's. A novel company called SAMPLION Ltd., grounded in May 2009, starts to deliver PMEU applications for different areas of microbiology.

Main features of PMEU enrichment, assay and testing procedures are its capability to enhance the growth of target microbes, its sensitivity and the challenge it serves when simulations of real target environments are needed. These features have already been proven in several research projects, derived eg. in hospitals, environmental laboratories and paper industry institutes.

We shall present these applications and refer our scientific articles, based on clinical, environmental, biotechnical and paper industry research projects, in this blog. All writers are experienced PMEU users, having long careers in microbiological research.

Thursday, July 9, 2009