PMEU as a tool for biofilm testing

PMEU Method was presented in PIRA Paper Industry Symposium, Barcelona, in October 2009. The basic PMEU model can be applied to diverse test types which help to construct biocide programs for the prevention of sessile and biofilm growth of bacteria in paper machines.

The picture above shows a typical test situation where process water sample, biocide and test coupon (made of steel) are installed in a PMEU syringe. Prevention of sessile growth can be monitored with ATP Assay, biofilm growth with UV Epifluorescence Microscopy.

In addition to the basic PMEU model, the novel PMEU Spectrion which measures the turbidity of all ten samples automatically, can be applied to any microbiological growth / growth prevention test of liquid or slurried samples from paper manufacturing processes. This device can handle also relatively turbid samples because it stops the mixing of the samples before turbidity measurement, allowing heavy particles (like mineral pigments) to sediment and enables therefore the measurement of bacterial cloudiness of the sample.

Competition between tube and colony count methods

All began with beef broth.

Dr. Louis Pasteur invented this method for the cultivation of diverse microbes. The famous "Swan Neck" trial was also performed with beef broth.

One of the first solid media for microbiological cultivations was the surface of a potato, presented by Dr. Robert Koch.

Colony counts began to be more and more favored by microbiologist because the colonies gave a chance to the immediate isolations of strains. The visual appearance of colonies on solid agars also help to identify the actors of the play, the species of a sample. Membrane filtration method also rise the popularity of colony count method, as well as the relatively good accuracy of colony count analyses, compared to the broth methods.

The role of the tube methods, however, has turned to be more important today. The limitations of the colony count method, correlated with the features of the samples (turbidity, toxic compounds etc.) and the slow growth (compared to the broth cultivation), are obvious. Testing of growth-affecting compounds like biocides are also easier and more reliable to perform in a solid media. When testing of those agents shall be done in the real environment (like the process water of a paper machine), the only alternative is the tube test. Detection of the response of stimulating and inhibiting agents can be done with various methods (photometry, colorimetry, turbidity, ATP Assay etc.) easily. Quantitative analyses of microbial counts can also be performed much faster with a (MPN) tube method than with the colony count method.

As a conclusion: colony count methods suit very well for purposes like the counting of CFU values as well as the selective cultivations of the total population to detect certain microbial groups. Testing of the effects of diverse growth factors (temperature, pH, biocidic and biostatic compounds etc.) should be performed with the tube methods, however. Growth on/in a solid medium does not correlate with the growth of the population in its original environment. Biofilm trials shall always be performed in liquids, never on solid media.

Various analytical tools have been developed for the measurement of the growth responses (pH, turbidity, impedance) automatically from the tubes and the most novel method, PMEU "mini-fermentor", gives the chance to perform all tests with the highest speed and - if needed - in the original samples to simulate the real growth environment of the microbial population. This method will be presented in PIRA Paper Conference, Barcelona, in next October.

A new PMEU application: quantitative MPN analyses of microbial counts

PMEU method is based on the rapid cultivations of several samples. The old idea to apply it in MPN (Most Probable Number) analyses has now proven to be correct: referring the Finnish Standard Book "SFS-KÄSIKIRJA 94: Mikrobiologiset vesitutkimusmenetelmät" (Methods for Microbiological Water Analysis) and discussions with specialists, PMEU can be used as an alternative, rapid method instead of the traditional technique, tube series in water bath or in an incubator. PMEU itself works as an incubator with a temperature deviation of < 0.1 oC.

A combination of 4 (levels of dilutions) * 5 (repeats) allows to follow the Finnish standard SFS 4447 (The Tube Method in Microbiological Water Analysis) as well as standards derived of it like SFS-EN ISO 9308-3 (for and coliforms) and SFS-EN ISO 7899-1 (for enterococci). Standards usually give MPN tables in the framework of 3*5 tubes (eg. for dilutions from 0 to 0.01) but PMEU gives an extra level (eg. 0 to 0.001) which covers a wider range of microbial counts. Samples with unknown levels of microbial densities are therefore easier to analyse correctly.

It seems that the leading status of membrane filtration has revised today. There are types of samples which are difficult or impossible to analyse with them (too much suspended solids etc.) and tube tests like MPN should be chosen. PMEU Tube Tests should be preferred also in situations where fast results (in hours, compared with days with colony count analyses) are needed.

The microbiological control of certain paper industry samples (pulp slurries,starches, minerals) are better to perform with tube methods. An example of the priority of the tube methods can be seen when samples with polymers should be analyzed: polymers tend to stuck membranes immiadely but do not prevent any analyses performed with tube methods. Rapid detection of harmful or hazardous bacteria can also be done faster with selective broths than in/on selective agars.

Applications of PMEU method for biofilm research and testing of biocides against biofilm growth

Paper machine biofilms have been studied already several years with PMEU method by IM. Test coupons, made of steel brands used in paper machines, have been installed inside PMEU syringaes and the growth has been observed with UV Epifluorescence Microscopy after a short incubation period (see picture above).

This technique has now been modified for ordinary light microscopes, too. Steel coupons have been replaced by specified glass slides and the Gram-stained biofilms can be observed with Bright Field Microscopy - no expensive epifluorescence microscopes are needed in this application.

This method will detect all biofilm-producing microbes and testing of biofilm-preventing biocides is also possible simultaneously. Primary attachers typically appear on the slides in just hours and mature biofilms are available in 12...24 hours. This application is very suitable for all areas of industry where the hygiene of surfaces is important. It can also be applied in every environmental research projects where the formation of biofilms in natural water environments is the subject of the study. Hygiene control of public swimming pools etc. also benefit of this method.

Sulphate Reducing Bacteria in paper industry.

SRB is an interesting group of bacteria which can use sulphate as an electron acceptor for the respiration. Despite the chances of certain other bacteria to use sulphate as the sulphur source for their S-containing cell components, the "real" sulphate reducers transform SO4(2-) to S(-2) in their energy metabolism and oxygen actually inhibits their growth - they are therefore obligate anaerobes. Certain yield of energy may be achieved through fermentation by SRB's but this type of metabolism is regarded as relatively insignificant one for them.

These bacteria have first detected in waste waters of sulphite pulp mills but modern paper machine processes can also induce their growth if certain sulphur-containing compounds are available.

In addition to H2S production (which is a hazardous gas), colour problems can arise because the metal sulphides. FeS is an indicator compound in the analytical detection of SRB's but also a harmful agent of discolorization of paper and paperboard. Last but not least, SRB's have been shown to be conneceted to a certain type of iron corrosion and it is all possible to find those problems still today whenever technical structures with poor steel quality and certain types of organic deposits on their surfaces are combined.

SRB's have also other, peculiar features like the tendency to follow non-exponential growth curve. They have been the subject of firm microbiological research only since the middle of 20th century because their need of anaerobiosis was not understood earlier.

FINNOFLAG Ltd. is currently developing a novel method for the sensitive detection of SRB's with PMEU method - more about this topic in next posts.

The need of bacterial identification?

IM has discussed about alternative methods for the detection of hazardous or harmful bacteria with Dr. Elias Hakalehto.

It is most important to know the pathogens which will appear in patient samples. Clinical microbiologists shall know who are the enemies of the ill people: their metabolic capabilities, antibiotic resistence patterns etc. Their overall features are easy to find from literature or internet whenever the name of the species is known. This identification can be performed by selective cultivations on agar plates or in PMEU incubator, and further tests like microscopic examinations, API ID systems, immunological tests and/or PCR can be done to confirm the basic identification.

Paper mill is definitely another challenge for microbiologist. In some (relatively rare cases) the names of microorganisms are important to know: if the product shall have high hygiene quality (like LPB and other food-grade cartonboards) or questions about bioterrorism have been arisen (spore-forming Bacillus anthracis as an example). The occurrence of Legionella pneumophila is also a risk in the waste water treatment of paper industry today. Selective cultivations, either on plates or in PMEU, are the solid solutions for continuous microbiological control in those cases. PMEU is preferred because its speed (hours, compared to days with colony count analyses).

Papermakers shall focus more on the metabolic activities than the names of bacteria which they are living with in paper mills, however. Continuous inoculation of the paper production processes by contaminants, delivered with incoming lots of starches, mineral fillers, raw water, dry pulp etc. shall be controlled to avoid spoilage (amylolytic activity as an example), biofilm and slime growth, tastes and odours, spots and colours in the product etc. Because the wide range of bacterial species and their origin from the nature itself, clinical methods do not suit very well for this monitoring. There is no time to start labourous cultivations, pure cultures and identifications when the bacterial input continues day and night, "7/24". PMEU seems to be an excellent tool to check the basic features of process populations, their biocide resistence patterns included.

One important fact must also be taken into account. There are a lot of harmful microbes which actually cannot be cultivated on agar at all. One example are certain filamentous bacteria which may cause biofilm layers into the processes. They can be cultivated in some broths, however, but the usage of the original samples as the growth medium is the best way to detect them all. This can be done with ordinary mb laboratory equipment or with PMEU incubator.

Identification of bacterial species is still needed when the mapping of contamination routes into the processes is the subject of the study. IM will discuss about the microbiological mapping in his next posts.

Statistical methods in microbiology.

IM has discussed about the evaluation of novel microbiological methods with several professionals. His knowledge of statistical methods in microbiology bases on the lessons by Prof. Seppo Niemelä, who was (and still is) a well-known specialist in this not-so-well-known area of microbiology.

Testing of microbiological data is more complicated than similar analyses in chemistry. The main reason is the model of repeat distribution: chemistry follows the ordinary normal distribution but the colony counts of microbiological analyses are featured by the Poisson distribution. The reason for this difference is easy to understand: the count of molecules is overwhelming when compared to the limited count of colonies in microbiological cultivations. The dependence of variance on the mean of the data is another problem of colony count analyses, preventing the usage of parametric methods.

Luck enough, there are some non-parametric statistical analyses for Poisson-distributed data, helping the comparisons of means and trends of colony count results.

A novel problem seems to have arisen in microbiological evaluations since 70's. Many modern, automatized instruments of microbiology are not based on the measurements of chemical concentrations or counts of colonies but on specified metabolic activities of microbes. Examples of these analytical procedures are eg. measurements of impedance, turbidity, pH or CO2 production. Because these parameters are in a close connection to the growth rates - and to a new parameter, time - , their evaluations are very challenging procedures.

- More in next posts...

Identification of bacterial species.

Questions about the identification of certain bacterial species are rising time after time. Paper mills - and their customers - are interested to know if the product, paper or board, contains harmful bacteria like food-poisoning bacteria, or even hazardous ones. Could terrorists inoculate a paper machine with Bacillus anthracis? Could Listeria live in wet end processes? Bird-flu?

Another reason for the question is the safety of working environment. Are high densities of bacteria in the wet end air dangerous?

Fortunately all information of the environmental needs, growth factors and capability to survive dry end treatment (="pasteurization") show that there is no need to worry if those real pathogens could be encountered in paper industry products.

There are still some areas inside paper mill where the identifications are needed, like

* food-poisoning, sporeforming bacteria (most important: Bacillus cereus)
* bacteria causing raw material breakdown (amylolytic ones etc.)
* potential primary attachers, causing biofilm formation (several genus)
* bacteria which can cause health symptoms and diseases(Legionella pneumophila, certain coliform bacteria)

Identification of specified bacteria from wet end population isn't easy task. Mixed population, sample matrix, low number of target organisms, their distribution inside the machine all lead to very challenging task.

I will refer the traditional and novel methods for the detection of specified bacteria in next posts. Coming back...

Connections of paper industry microbiology to other sectors of microbiology: what is actually needed?

To make any definitions of paper industry microbiology, it makes sense to compare it with elder sectors of microbiology. Despite the microbiological problems of the paper processes and the paper products have been obvious since the beginning of machine-scale production of paper and board on 19th century, their effects have get worser when the scale, speed and raw material repertoir have increased during last decades. The tradition of the microbiological control, as well as the history of biocide research, intended in the "healthcare" of paper and board machines is therefore much shorter than in related areas like in dairy or food microbiology.

Paper manufacturing processes could be seen as ecosystems where several, complicated microbiological processes are continuing day and night. Microbial communities perform their important role as the actors of chemical transformations which shall modify most living and very many non-living substances into forms which will support the growth of other living creatures. Many species of immigrant bacteria, coming into the processes with the raw water, mechanical fibres and several additives, will feel fine: favourable temperature, pH level and nutrient concentrations, as well as good aeration and a huge supply of contact surfaces to build up biofilms, are available for them. They really do not make any difference between their lives outside and inside of the paper mill walls.

Unfortunately (not for the microbes but for the paper production) there are some features of paper machines which are similar with fermentor and bioreactor processes of biotechnological industry. So many growth factors (some of them were mentioned above) will be kept on so controlled levels that the adaptation of certain microbiological population cannot be avoided. It shall also be kept in mind that the long running periods will increase the microbiological risks by allowing long growing periods of microbes inside the machines.

How to control these problems?

Measures to dose biocidic compounds into the processes cannot be avoided because the conditions of paper and board machines cannot be adjusted on biocidic levels: the rise of the overall temperature over +80 oC is impossible, like the rise of pH value over 12. Before significant technical improvements to prevent the microbial growth in the paper machine processes could be done (if ever), the biocidic treatments and their rapid control methods like ON LINE biofilm measurements and frequent (at least once per 8 hours) AT LINE microbiological control of the main contaminating routes, wet end processes and towers containing white waters, pulps and brokes are the most important tools to secure the runnability of the machines and the quality of the products.

With the price of only 2-3 jumbo rolls can reliable instruments for the AT LINE microbiological control of the wet end processes be bought today. Alternative methods, many of them representing molecular biology methods, are available, but those which can show not only the counts of certain species but also the overall metabolic activities of the waterborne microbes and their potential to produce biofilms should be preferred. A combination of PMEU incubations and ATP Assays, with the addition of PCR if needed, is the most recommended procedure to show the effects of biocides on the microbial activity. PMEU method can be applied to biofilm testing, too.

The most important thing is that not only the counts of microbes (how high they may ever been) but also their overall metabolic activity and certain actions like breakdown of starches by amylase enzymes or production of H2S and H2 in anaerobic conditions shall be controlled all the time when the machines are running. All the laws of microbial ecology are present both in the nature and inside the machines - and they can lead to severe problems if counteracting does not work.

"Top Three" microbiological problems of paper machines

Certain types of microbiological problems in paper mills seem to be acute all the time. Looking back to last months, this may be "Top Three" among them:

* Microbiological spoilage of raw materials. This is an everlasting hazard for mineral and starch slurries, and the reasons are very easy to understand: both raw materials mentioned may contain high densities of bacteria (mainly aerobic sporeformers and actinobacteria), slurries containing starch are very nutritive growth media for different microbial species and the very challenging biocidic treatments of slurries (especially mineral ones), when inaccurate, can lead to the total spoilage of them.

* Growth of biofilm and production of slime. This problem seems to be connected to poor washing and boil-out programs which leave rests of biofilm inside the machine and give growth time for it because too long running periods. The chose of ineffective biocide and/or its insufficient dosing can also stimulate the activity of these trouble-makers.

* Microbial growth in the broke system. Especially big machines with large broke towers suffer of this problem. If the basic biocide program is insufficient and the retention times inside the towers are too long, aerobic population tend to increase the number of its cells to the level of 10 000 000 cfu/g or even higher. Consumption of oxygen by respirating bacteria leads to anaerobic conditions, redox potential will be dropped and the growth conditions for both fermentative and anaerobic bacteria turns to be excellent. Drop of pH, slime and spore formation, smells and odours - even the production of H2S and H2 - will be found in such situations.

There are some measures to prevent these hazards. Growth period of microbial population shall be kept as short as possible, the control of waterborne and bioflim bacteria shall be as rapid and frequent as possible and the bioside programs, intended in killing of raw material, process water and biofilm bacteria shall be evaluated more frequently.

A realistic and accurate way to control both process water and biofilm growth, as well as to evaluate biocide programs, is now available. The Finnish company SAMPLION Ltd is manufacturing and selling "Portable Microbiological Enrichment Unit", a "mini-fermentor" for 10 simultaneous tests in controlled conditions, to detect the failures of biocide programs in only hours (watesr) or days (biofilms). Some results of PMEU's paper industry applications will be published in next Spring - coming back to refer them later.

Some wrong ideas about the overdosing of biocides will also rise up frequently among publicity. Basically it is not a question of only the cubic meters of biocides consumed, however, The chose of most effective biocides for different areas of processes towards different problems, the dosing of these compounds, their type of action and some other factors play a major role when building an effective biocide program for paper machine. Overdosing of biocides is a problem only in cases, when the program does not work, and leads to the loss of money and the rise of biocide concentration in paper machine effluents.

What will the next Spring bring to Finnish paper industry?

After a most busy period in May 2009, IM would like to write something (partly because his insomnia - a common symptom during light & bright Nordic summer nights) to the readers of his blog.

Several seminars for the rise of Nordic forest industry have arranged and will be arranged before Finland "will be closed" for summer holidays. - This is, by the way, a common feature of Finnish lifestyle, and easy to understand: you shall first work in dark and rain for months, the temperature drops well below 0 oC, and you should try to be active all through the winter. But when the summer comes, everybody will wake up again and "see a light at the end of a tunnel" (which was the title in the annual meeting of Finnish Paper Engineer's Association). And then, during the summer months, citizen tend to move to their summer cottages and villas to just rest and collect forces to tolerate the next winter..

Back to the topic:

There was a certain drop in the number of visitors, speakers and companies with exhibition desks in the annual Congress "BioChem" in Helsinki, 27.-29.5.2009. It was a pity to see that a lot of paper industry-associated companies did not arrive to show their products. The depression of global economy was very easy to sense. After all: there were still active participants left, eg. the novel company SAMPLION Ltd. which is now distributing PMEU system for all areas of microbiological control.

Positive attitudes were obvious also in the annual "Summer Summit" by AEL, the major education company for adult professionals, paper industry ones included.

What will happen next? A conference, discussing of future forecasts of forest industry, will be held in the middle of June in Jyväskylä, Finland. After that the annual PIRA congress will be held in Barcelona, Spain, in October.

If only the paper industry research will be activated again..It is, of course, a question of money, but, after all: to leave brilliant Finnish researchers without work is really a vast of excellence. It has be proven several times that the rebuilding of research groups after a depression is a much harder work than to start those activities from zero. It would therefore be better to keep such organisations in work over the bad periods.

IM is looking forward to the next Autumn! A most interesting thing to see what are the forecasts of Nordic P&P industry when the summer is over!

What's new?

Last weeks have been very active in IM's life.

Annual meeting of Finnish Paper Engineers's Union (Paperi-insinööriliitto) has given some new opinions about paper industry in our country. Optimistic views were mainly honest ones, and the title of the meeting indicated better views into the future: "There is light in the other end of the tunnel"!

Training of paper industry employees seems also to be continued. AEL (biggest professional training coalition in Finland) kept a course of paper industry sampling a couple of weeks ago in Lappeenranta, and AEL and its smaller "competitor", Finn-Fiber Oy (especially focused on paper industry issues) are planning new courses to be kept already in next summer and autumn. It is very interesting to join the planning groups of these companies and be with when finding current topics and speakers for seminars and lectures.

Main issue, of course, are the measures to spare money, and in this context, it means better drive of the machines. Better sampling and faster analyses of the processes are most important ways to prevent all kinds of process problems and product faults, and the role of PMEU (Portable Microbiological Enrichment Unit) seems to rise even more: a new coalition of companies called Samplion Oy has started the mearketing of this microbiological analyse tool, and it will be presented on Chembio Fair at the end of this month in Helsinki, Finland. We'll meet by the stand of Samplion Oy!

Negative issues should also be mentioned. One of them is the market situation of Russian short fiber: Carelian companies shall fulfill their task to collect certain amount of birch logs, no matter they cannot sell them to Finland because high taxes, set by Russian government. IM wishes good luck for the negotiations about this problem!

We are living interesting times - let's see what happens in the area of paper industry in next months!

Most important issues in paper industry microbiology?

Referring the responses of the readers of IM blog, it seems that the most important issues of this QC area are:

* Klebsiella pneumoniae (and other pathogens)
* Activated sludge
* Economical effects of mb control for the mill

For some reason, no such interest has been focused to

* biofilms
* product hygiene
* raw material storage
* HACCP in P&P industry
* novel mb methods

IM is very pleased for the active visits on his blog page but, after all, will also be very pleased to hear questions about the issues mentioned above.

Looking forward to hear your questions,

IM

Annual Award "Forest Industry Trainer 2008" by AEL

Microbiology has regarded as a considerable area of paper industry support by Finnish training institute AEL a couple of days ago.

This award was definitely an issue of personal satisfaction for IM. The fact that the role of microbiological control of paper mills has been taken into consideration is the most important thing, however. Courses and lectures of P&P microbiology will focus the attention of personnel in this industry to the significant spares which are resulted by better maintenance of raw materials, run of the machines and quality of the products.

This award was given in the annual meeting of PI, Finnish Society of Paper Engineers. The forecasts of speakers in this meeting were optimistic ones and opinions seem to be pretty positive ones. Funding of several paper technology students also indicates these feelings. The final conclusion of this meeting can therefore be: P&P industry is not a "sunset branch" of global economy, no matter severe economical problems have arisen during last years. Paper and board is still needed worldwide, as well as new fiber-based bioproducts. Feelings after the meeting were therefore very good, indeed.

Five most important issues in current P&P industry?

IM is wondering, what will the future of P&P be like.

Referring his ideas, five major topics are needed:

1 PRODUCT: you shall have some product which is really needed among customers. An example: LPB (everybody will buy milk and juice in carton packages every day during her/his lifetime).

2 PERSONNEL: persons on all levels of the company, interested to sell good products.

3 MARKET: the only question: are the customers well aware about your product? If not, why?

4 RESEARCH: R&D in P&P is not a region of "sunset". A lot can be done to improve the quality of products, production etc. It would be possible to find even new products: why not to sell motor oil in LPB containers?

5 DEVELOPMENT. This is the most difficult issue. R&D is interested to develop new ideas but the economists do not see any forecasts for the innovations - why?

- O.K. IM is just a microbiologist and do not understand the global economy. He is still asking, however: where is the development and optimistic forecasts in P&P industry?

IM will also ask: are the Asian people more enthusiastic to develop P&P industry? Not only economically but also in their minds?

Control of activated sludge process in paper industry.

Effluents of pulp and paper mills are relatively simple to treat by activated sludge process - in theory. Some severe problems have arisen in practice, however.

Temperatures of incoming waste waters have risen during last years. The most obvious reason for this phenomenon is the increased recirculation of process waters inside the mills. Poor sedimentation of flogs, caused by the heavy growth of filamentous bacteria, has also become to be more and more hard problem for purification plants. There may be some kind of correlation between temperature and bulking of sludge?

Another problem, potentially very hazardous, is the growth and recirculation of Legionella pneumophila inside activated sludge process. The role of Protozoa has now been established: they act as carrirs of bacteria backwards in returned sludge.

Dosing of nutrients into biological treatment steps isn't any hard task anymore because the need of phosphorus and nitrogen has been studied very deeply, and simpe AT LINE testing for the optimization of them is easy to perform in the mill labs.

Two problems, mentioned before, still exist and need R&D activities to be solved: bulking of the sludge (by filamentous organisms) and L.pneumophila.

Transfer of microbiological control from institutes to mill labs

Rapid development of analytical microbiology has been obvious during last 20 years.

After the beginning of IM's career in paper industry (est. 1982) a significant increase of novel methods has taken place. Slow and labourous colony count analyses have been replaced with novel, advanced methods in certain laboratories on 1990's.

Biomass, surface hygiene, condition of activated sludge, biofilm formation - among even more subjects - can be assayed by luminometric methods today.

Light and UV microscopy is another basic tool of paper industry microbiology today. Very valuable results have been achieved with TEM on 1980's and articles about sporeforming bacteria and biofilms, based on electron microscopy, have published by researcher all over the world. TEM is, however, such an advanced research instrument which is practically impossible to apply into everyday microbiological control of pulp and paper mills. Confocal microscopy has given brand new ideas about the structure of biofilms but it is also a too complicated method for mill labs. In opposite, light and epifluorescence microscopy aren't too expensive; they definitely need a lot of training for the personnel which is no big problem, however: in Finland (and IM is sure, in other countries, too) are training companies who will have annual microbiology courses for paper industry under titles like "Paper Industry Microscopy" and "Methods for Process and Product Hygiene in Paper Industry".

Tools of molecular biology have replaced the previous generation's major tool, FAME (Fatty Acid Methylated Esters - an application of gas chromatography to perform identifications of bacteria). But the limits of PCR and similar methods are obvious: they cannot show what is really happening inside the machines! They only give - valuable, of course - information about microbial species but do not explain and forecast those metabolic reactions, succession of population, risk of biofilm formation etc. which are more important for the drive of machines, good housekeeping of raw materials and product hygiene.

Basic methods of modern microbiology like DEFT, ATP Assay, PCR and other should therefore be combined with simple simulators, driven in mill labs. This is already possible: the first system for this target, PMEU (Portable Microbiological Enrichment Unit) has been tested and used by IM since the beginning of 2000's and it has been proven to be a most valuable tool for rapid raw material, process and biofilm studies today.

An ecological point of view shall be applied to everyday mb control of the pulp and paper mills. This question is discussed in the article "Paperikone - ekosysteemi ja bioreactori" ("Paper Machine - An Ecosystem and A Bioreactor") by JM in the annual of Finnish Microbiology Society (INOCULA 2007 / 1 - unfortunately only in Finnish). Understanding of these two natures of a paper machine gives new chances for the mills: it gives the ability to forecast microbiological events inside the processes and it also give extra time to prevent problems.

Challenges of Biotech

IM will refer the excellent book "The Biotech Investor" by Tom Abate (Owl Books, 2003, New York) again. IM also tries to reserve his time to read it through, no matter it is a dense collection of biotech and marketing facts.

In the preface of this book Tom Abate says:

"Biotechnology is an experimental field. Only a handful of of biotech companies currently have products on the market. The vast majority of biotech firms run at a loss during long developmental cycles aimed at proving their founding thesis. Experiments take time, and they often fail. Even when experiments have been done, and their results analyzed and presented at scientific meetings, biotech medicines undergo an excruciating process of regulatory review the can lead to approval, rejection or demands for more testing...In biotech time, development horizons strech for years, even decades."

This is familiar for every modern biotech researchers. Dr. Abate refers medicine research but relatively similar problems may arise when a novel microbiological analyse method is the subject of the research. Approval is then depending not on the health of the customers but the potential markets (IM's opinion). IM still thinks that such a development task can be fulfilled in only years, but marketing may be a challenging task, indeed.

Contacts between paper industry personnel and biotech researchers shall be developed. There are other alternatives than the traditional fairs and exhibitions: discussions in Internet blogs, nings etc. would help significantly to present all modern microbiological tools and evaluate their use in paper industry microbiology. Unfortunately the main problem, time, still stays, even gets worser: people tend to be in a hurry in their working life today. To reserve a period, even short, to investigate new methods will still be worth to do!

Good News' Day?

It is now published: TV news told tonight about the fusion of KCL (national pulp and paper R&D center) and VTT (national center of technical R&D) in Finland.

As an insider for several years, IM has seen the synergy of these two institutes and their university partners. Now, as an outsider observer, IM was slightly surprised but, after a moment of wondering, he understands the basic facts which have led to this solution.

This new, big R&D center will work towards R&D solutions more effectively and economically, IM hopes. It also comes time to invite small, scientific enterprises to have cooperation with this major R&D unit. They will give fresh views on several areas like process control, hygiene and energy savings. They have been the origin of new ideas in several cases during IM's career, indeed.

IM is therefore looking forward optimistically! "Small can be beautiful"!

The need of paper machine biocides?

IM is wondering: are there any other sector of process industry, where microbes are allowed to grow like in paper industry?

Biotechnological processes, of course. But their populations are carefully selected and controlled. And they are employers of the company, not criminals trying to cause harm to the company.

Some significant changes in paper industry processes have taken place after the rise of active environmental care. Both the closure of water circulation and the cancelling of biocides having mercury as an active incredient are favorable for the environment, of course. But the microbial growth inside the paper machines has activated at the same time.

The beginning of neutral paper production led to an "ecocatastrophe" inside paper machines. The rise of pH value (as well as the rise of temperature, caused by the extended recirculation of water) was fatal for slow, acid-loving fungal growth. New mineral additives are an important source of certain types of bacteria, causing severe problems like production of slime, spoiling of the process compounds and hygiene faults of the products.

IM has published an article "Paper Machine: an Ecosystem and a Bioreactor" (INOCULA 1/2007. Helsinki, Finland). Many readers agree: paper machines offer ecological niches for bacteria, and the controlled environments of wet end circulations are very much similar with those of biotechnical processes, based on chemostatic fermentors.

A lot could be do to make paper machines more unfavorable growth environment for microbial contaminants. Very good results have been achieved in some projects where ecological aspects have been taken into account. These issues will be discussed later in this blog.

But the main question is: how much biocides we still need to control the microbial growth in all regions of a paper machine?

Incoming raw materials, sorry to say, may be very contaminated: the highest value of total count during IM's career has been over 100 000 000 cfu/g in a mineral slurry (which was fortunately replaced by a fresh lot by the supplier!). It is therefore obvious that a continuous control - both analytical and practical - is needed for starches, mineral pigments and other contaminated raw materials. This does not mean that all lots are spoiled: there are suppliers which know their response to deliver pure products to the mills but all kind of errors in biocidic pre-treatment, transport and storage of these products may happen.

Certain sites of paper machines also need biocidic treatments all the time. Chosing proper solutions for biocide programs (type of biocide, active compound, dosing sites, timing etc.) of a paper machine is a challenging tasks. In best cases, both the paper mill and biocide personnel are sitting down and discussing of the individual problems of the paper process hygiene.

When specified laboratory services, having tools like PMEU and biofilm microscopy, are included, the final result can be optimal one. Paper industry microbiologists can also help significantly by declaring the effects of process parameters on the growth of planktonic and biofilm bacteria.

The more competence is included, the better solution will be find.

Checkpoints of a paper machine.

Imagine yourself as a bacterium.

You are coming to the paper mill - maybe waterborne, maybe as a bug or as a spore in a lot of starch or mineral pigment.

What will happen to you?

Do you find an ecological niche inside the machine?

I am sure, you will find.

You will find a wet surface, some of your genes will be activated for the living inside a biofilm.

You have fellow bugs there. You could even communicate chemically.

And, after a period of lush life inside the biofilm, you get older and will be released into the water current.

And you wish, that there are no detectives, controlling your movements inside the paper machine: wet end systems, pulp and broke systems, mineral slurry and starch size routes...

The detectives belong to HACCP. They know where you will hide...

- This is my dream. Tools for this kind of HACCP are already available.

Some practical views into the future of biotechnology.

I will come back to the future of biotechnology.

The economical figures of traditional industries seem to be awful. We will see, what is the condition of such branches like paper, automobile and communication enterprises in 2010.

The optimistic view into biotechnology, presented by Osmo Kuusi in 1991, should be reviewed soon.

Another positive attitude has been presented by Tom Abate in "The Biotech Investor", published in 2003 and found by me under a layer of dust in the library of Jyväskylä University (Tom Abate 2003. The Biotech Investor: how to profit from the coming boom in biotechnology. Owl Books, New York).

Tom Abate tells about sectors of biotechnology like the previous authors, discovering this area. He calls them "The Genomics Wave", "The Proteomics Wave" , "The Biotech Toolmakers", "Green Genes", "The New Factories", "Brave New Worlds" and "The Biotech Clusters".

What these titles include, I'll try to explain whenever I have red this interesting book.

Before that, I have good news:

The leading newspaper in Central Finland, Keskisuomalainen, tells about the manufacturers of small-scale waste water treatment plants. Finnish legislations says that all small houses shall update their ww treatment systems before the end of 2013.

The count of these households is est. 300 000 in Finland!

Not to name the individual enterprises, there are a couple of small companies manufacturing/importing batch-type activated sludge systems in a small scale in our country. Two larger companies, having Finnish background, also exists.

Novel ideas to control the effluents of these mini-plants already exist, but the questions of evaluation, standardization etc. will have a sever effect on the application of these methods.

I think that it is now time for the Finnish government to accept these analytical methods to be applied by small-scale treatment plant producers.

Colony counts, developed in the beginning of 20th century, are valuable. But what we are needing today are the rapid methods for the control of both activated sludge itself and the effluents of these equipments.

Consulting in 2009?

I am starting my working again after a several days' flu by reading "The Paper Index Times". Updating the current situation in P&P is easiest by this way.

The columns by Jim Thompson, writing "Nip Impressions" (see http://news.paperindex.com/category/Blogs_Columns) are always very interesting. I would like to refer a chapter of his last issue here:

You may think it is self-serving (read: old Jim does not want the competition) to tell you to avoid consulting, but my real reason for saying this is consulting, unless you have some very special well known skills, is a daunting business to enter and be successful. I tell people coming to me seeking advice on this subject the following: If you like to write, and can do so quickly and expertly, and you like to be on job interviews, come on in, the water's fine. General consulting consists of two activities: writing and interviewing for your next assignment. It is hard work. On the other hand, if you have a particular exquisite skill and are well known, you can be a success. I once knew a gentleman capable of making any old winder run, no matter how decrepit. He was in high demand, so high of demand he decided to double his rates to see if he could reduce his workload. All he did was double his income. If he had thought, there was a hint of his "problem" staring him in the face: when the mill I worked at needed him, corporate would send their private jet to fetch him, no matter where he was, and bring him to our mill. This is a reputation you can retire on.

This is something which I have wondered over a period of several years.

Consulting, performed by individual specialists or small, private institutions would be very favorable for big companies today. As discussed several times in my blog before (and is worth to repeat) is the challenge of big P&P companies to have benefit by buying rapid services from independent sector of R&D.

All the sad news about closing R&D centres give an impression of fatal view ito future: nothing can be developed anymore. Research is all in vain. But this is not the truth.

There are always ideas growing on among researchers, no matter economical situation is alarming. And it is easy to understand: researchers have adopted a positive view also in problem situations during their studies and career. Just thinking about trouble shooting of machines which would not run: you will continue your task until you find some solution. This IP is something which should be valuated and used today, no matter your "Profit 200X did not have any significant success.

If you are working for P&P, you have to be optimistic. Otherwise you do not survive.