Doctoral dissertation of Teemu Rinttilä, senior research scientist at Alimetrics

Title: Real-Time PCR - A Molecular Approach to Investigate the Role of Intestinal Microbiota in the Pathophysiology of Irritable Bowel Syndrome

Time: 8.4.2011, at 12 noon

Venue: University of Helsinki, Viikki campus (EE-building, Walter-auditorium, Agnes Sjöbergin katu 2)

Opponent: Docent Petri Auvinen, University of Helsinki

Supervisor: Professor Airi Palva, University of Helsinki

Electronic version: https://helda.helsinki.fi/handle/10138/25853

Protocs employs a multi-disciplinary team of electrical and mechanical engineers and scientists, who share a passion for learning-by-doing and enjoy finding novel solutions. The Protocs team specialises in providing custom design, manufacturing and assembly services for various fields related to life sciences. Our services are offered as projects that effectively capture the innovativeness of the entire Protocs team. We have a strong background and experience in the laboratory environment and R&D activities, but we are always willing to challenge our skills in new areas.

Quantifire, a subsidiary of Alimetrics Ltd, has acquired new facilities adjacent from Alimetrics’ laboratory. At present Quantifire runs over 100 000 qPCR reactions annually. Extensive laboratory work from sample handling and DNA extraction protocols to data analysis and statistics requires specific expertise. We have decided to commercialize this expertise under brand name QuantiFire and offer service packages to our industrial and academic clients.

7-12.3.2010

Suvi Ojanperä has presented a poster at the Manfred Donike Workshop, the 28th Cologne Workshop on Dope Analysis concerning the characterization of microbiological contaminants in urine. Contamination of urine samples by microbes may result in complete or partial degradation of endogenous substances, drugs and their degradation products. Certain microbes are able to alter the steroid profiles by synthesizing or degrading steroids or by hydrolyzing conjugated metabolites. This may lead to false positive and negative results, being a threat for reliable doping control. We have carried out a sequencing study for revealing microbial groups which are present in authentic doping control urine samples and can potentially alter the steroid profiles.

Management of intestinal microbiota of monogastric animals has increased in importance since the ban of growth promoting antibiotics in many countries. Organic acids have been used as alternatives to antibiotics by many feed manufacturers. Regardless of the wide usage, the effect, dose response and mode of action of acids on intestinal microbes is poorly understood. In this study, we investigated the effects of dietary supplementation of three commonly used products, namely formic acid (FA) (90%), DL-methionine (DLM) (99%) and liquid methionine hydroxy analogue-free acid (88%), on ileal microbiota of pigs. Laboratory simulation system, mimicking swine ileum, was used to study the products at various concentrations and combinations. Furthermore, selected combinations were tested in a piglet trial to confirm the findings made in in vitro studies. FA turned out to have a dual effect on ileal microbiota. At concentrations below 0.5%, it significantly stimulated bacteria, but at higher inclusion rates it was highly inhibitory. This finding, which was consistent in in vitro and in vivo studies, implies that reducing the dose of FA does not lead to a diluted inhibitory effect, but in fact, an opposite, stimulatory effect on intestinal microbiota. It is highly important that feed compounders acknowledge this finding. Unlike FA, the inhibitory effect of methionine hydroxy analogue on ileal bacteria was linearly dose dependent and significant at inclusion levels above 0.2%, in vitro. Partial replacement of methionine hydroxy analogue by FA, or FA by methionine hydroxy analogue, led to an unpredictable outcome due to the dual effects of FA; e.g., a minor inclusion of added FA changed the inhibitory effect of methionine hydroxy analogue into microbial stimulation by FA. Inhibition of ileal microbiota by methionine hydroxy analogue was detected only in in vitro studies, suggesting that intact methionine hydroxy analogue may not have reached the ileum, in live animals. Therefore, if the target is to ensure the inhibitory effect of FA, the FA level in feed should be kept above 0.6%, and not reduced, if methionine hydroxy analogue is used as a methionine source instead of DLM. DLM was totally inert with regard to bacterial growth and metabolism, both in vitro and in vivo. The results of these studies reveal the importance of knowing how each acid product works. Inconsistent results in animal trials may have been partly due to quadratic dose–response effects of products, and unpredictable product combination effects.