Bacterial community changes in copper and PEX drinking water pipeline biofilms under extra disinfection and magnetic water treatment.

AIMS: To study the stability of biofilms and water quality in pilot scale drinking water copper and PEX pipes in changing conditions (extra disinfection, magnetic water treatment, MWT). METHODS AND RESULTS: Next-generation sequencing (NGS) of 16S ribosomal RNA genes (rDNA) to describe total bacterial community and ribosomal RNA (rRNA) to describe active bacterial members in addition to traditional microbiological methods were applied. Biofilms from control copper and PEX pipes shared same most abundant bacteria (Methylobacterium spp., Sphingomonas spp., Zymomonas spp.) and average species diversities (Shannon 3·8-4·2) in rDNA and rRNA libraries, whereas few of the taxa differed by their abundance such as lower total Mycobacterium spp. occurrence in copper (<0·02%) to PEX (<0·2%) pipes. Extra disinfection (total chlorine increase from c. 0·5 to 1 mg l-1 ) affected total and active population in biofilms seen as decrease in many bacterial species and diversity (Shannon 2·7, P < 0·01, rRNA) and increase in Sphingomonas spp. as compared to control samples. Furthermore, extra-disinfected copper and PEX samples formed separate clusters in unweighted non-metric multidimensional scaling plot (rRNA) similarly to MWT-treated biofilms of copper (but not PEX) pipes that instead showed higher species diversity (Shannon 4·8, P < 0·05 interaction). CONCLUSIONS: Minor chlorine dose addition increased selection pressure and many species were sensitive to chlorination. Pipe material seemed to affect mycobacteria occurrence, and bacterial communities with MWT in copper but not in PEX pipes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study using rRNA showed that chlorination affects especially active fraction of bacterial communities. Copper and PEX differed by the occurrence of some bacterial members despite similar community profiles.

Short communication: Genomic prediction using different single-step methods in the Finnish red dairy cattle population.

Single-step genomic prediction models utilizing both genotyped and nongenotyped animals are likely to become the prevailing tool in genetic evaluations of livestock. Various single-step prediction models have been proposed, based either on estimation of individual marker effects or on direct prediction via a genomic relationship matrix. In this study, a classical pedigree-based animal model, a regular single-step genomic BLUP (ssGBLUP) model, algorithm for proven and young (APY) with 2 strategies for choosing core animals, and a single-step Bayesian regression (ssBR) model were compared for 305-d production traits (milk, fat, protein) in the Finnish red dairy cattle population. A residual polygenic effect with 10% of total genetic variance was included in the single-step models to reduce inflation of genomic predictions. Validation reliability was calculated as the squared Pearson correlation coefficient between genomically enhanced breeding value (GEBV) and yield deviation for masked records for 2,056 validation cows from the last year in the data set investigated. The results showed that gains of 0.02 to 0.04 on validation reliability were achieved by using single-step methods compared with the classical animal model. The regular ssGBLUP model and ssBR model with an extra polygenic effect yielded the same results. The APY methods yielded similar reliabilities as the regular ssGBLUP and ssBR. Exact prediction error variance of GEBV could be obtained by ssBR to avoid any approximation methods used for ssGBLUP when inversion left-hand side of mixed model equations is computationally infeasible for large data sets.

Diversity of ribosomal 16S DNA- and RNA-based bacterial community in an office building drinking water system.

AIMS: Next-generation sequencing of 16S ribosomal RNA genes (rDNA) and ribosomal RNA (rRNA) was used to characterize water and biofilm microbiome collected from a drinking water distribution system of an office building after its first year of operation. METHODS AND RESULTS: The total bacterial community (rDNA) and active bacterial members (rRNA) sequencing databases were generated by Illumina MiSeq PE250 platform. As estimated by Chao1 index, species richness in cold water system was lower (180-260) in biofilms (Sphingomonas spp., Methylobacterium spp., Limnohabitans spp., Rhizobiales order) than in waters (250-580), (also Methylotenera spp.) (P = 0·005, n = 20). Similarly species richness (Chao1) was slightly higher (210-580) in rDNA libraries compared to rRNA libraries (150-400; P = 0·054, n = 24). Active Mycobacterium spp. was found in cross-linked polyethylene (PEX), but not in corresponding copper pipeline biofilm. Nonpathogenic Legionella spp. was found in rDNA libraries but not in rRNA libraries. CONCLUSIONS: Microbial communities differed between water and biofilms, between cold and hot water systems, locations in the building and between water rRNA and rDNA libraries, as shown by clear clusters in principal component analysis (PcoA). By using the rRNA method, we found that not all bacterial community members were active (e.g. Legionella spp.), whereas other members showed increased activity in some locations; for example, Pseudomonas spp. in hot water circulations' biofilm and order Rhizobiales and Limnohabitans spp. in stagnated locations' water and biofilm. SIGNIFICANCE AND IMPACT OF THE STUDY: rRNA-based methods may be better than rDNA-based methods for evaluating human health implications as rRNA methods can be used to describe the active bacterial fraction. This study indicates that copper as a pipeline material might have an adverse impact on the occurrence of Mycobacterium spp. The activity of Legionella spp. maybe questionable when detected solely by using DNA-based methods.

Cow-specific diet digestibility predictions based on near-infrared reflectance spectroscopy scans of faecal samples.

This study was designed to obtain information on prediction of diet digestibility from near-infrared reflectance spectroscopy (NIRS) scans of faecal spot samples from dairy cows at different stages of lactation and to develop a faecal sampling protocol. NIRS was used to predict diet organic matter digestibility (OMD) and indigestible neutral detergent fibre content (iNDF) from faecal samples, and dry matter digestibility (DMD) using iNDF in feed and faecal samples as an internal marker. Acid-insoluble ash (AIA) as an internal digestibility marker was used as a reference method to evaluate the reliability of NIRS predictions. Feed and composite faecal samples were collected from 44 cows at approximately 50, 150 and 250 days in milk (DIM). The estimated standard deviation for cow-specific organic matter digestibility analysed by AIA was 12.3 g/kg, which is small considering that the average was 724 g/kg. The phenotypic correlation between direct faecal OMD prediction by NIRS and OMD by AIA over the lactation was 0.51. The low repeatability and small variability estimates for direct OMD predictions by NIRS were not accurate enough to quantify small differences in OMD between cows. In contrast to OMD, the repeatability estimates for DMD by iNDF and especially for direct faecal iNDF predictions were 0.32 and 0.46, respectively, indicating that developing of NIRS predictions for cow-specific digestibility is possible. A data subset of 20 cows with daily individual faecal samples was used to develop an on-farm sampling protocol. Based on the assessment of correlations between individual sample combinations and composite samples as well as repeatability estimates for individual sample combinations, we found that collecting up to three individual samples yields a representative composite sample. Collection of samples from all the cows of a herd every third month might be a good choice, because it would yield a better accuracy.

An extensive gastroenteritis outbreak after drinking-water contamination by sewage effluent, Finland.

An inappropriate cross-connection between sewage- and drinking-water pipelines contaminated tap water in a Finnish town, resulting in an extensive waterborne gastroenteritis outbreak in this developed country. According to a database and a line-list, altogether 1222 subjects sought medical care as a result of this exposure. Seven pathogens were found in patient samples of those who sought treatment. To establish the true disease burden from this exposure, we undertook a population-based questionnaire investigation with a control population, infrequently used to study waterborne outbreaks. The study covered three areas, contaminated and uncontaminated parts of the town and a control town. An estimated 8453 residents fell ill during the outbreak, the excess number of illnesses being 6501. Attack rates were 53% [95% confidence interval (CI) 49.5-56.4] in the contaminated area, 15.6% (95% CI 13.1-18.5) in the uncontaminated area and 6.5% (95% CI 4.8-8.8) in the control population. Using a control population allowed us to differentiate baseline morbidity from the observed morbidity caused by the water contamination, thus enabling a more accurate estimate of the disease burden of this outbreak.

Genetic analyses of metabolic body weight, carcass weight and body conformation traits in Nordic dairy cattle.

Improving feed efficiency in dairy cattle by animal breeding has started in the Nordic countries. One of the two traits included in the applied Saved feed index is called maintenance and it is based on the breeding values for metabolic BW (MBW). However, BW recording based on heart girth measurements is decreasing and recording based on scales is increasing only slowly, which may weaken the maintenance index in future. Therefore, the benefit of including correlated traits, like carcass weight and conformation traits, is of interest. In this study, we estimated genetic variation and genetic correlations for eight traits describing the energy requirement for maintenance in dairy cattle including: first, second and third parity MBW based on heart girth measurements, carcass weight (CARW) and predicted MBW (pMBW) based on predicted slaughter weight, and first parity conformation traits stature (ST), chest width (CW) and body depth (BD). The data consisted of 21329 records from Finnish Ayrshire and 9780 records from Holstein cows. Heritability estimates were 0.44, 0.53, 0.56, 0.52, 0.54, 0.60, 0.17 and 0.26 for MBW1, MBW2, MBW3, CARW, pMBW, ST, CW and BD, respectively. Estimated genetic correlations among MBW traits were strong (>0.95). Genetic correlations between slaughter traits (CARW and pMBW) and MBW traits were higher (from 0.77 to 0.90) than between conformation and MBW traits (from 0.47 to 0.70). Our results suggest that including information on carcass weight and body conformation as correlated traits into the maintenance index is beneficial when direct BW measurements are not available or are difficult or expensive to obtain.

Survival of Campylobacter jejuni in potable water biofilms: a comparative study with different detection methods.

Campylobacteria are important foodborne pathogens. C. jejuni bacteria have caused several drinking water-related epidemics in Finland. Normally, C. jejuni is not able to multiply in drinking water or in biofilms although it may survive in biofilms. The survival of C. jejuni in biofilms was studied using the Propella biofilm reactor. The number of bacteria was analysed with traditional culture methods and with fluorescence in situ hybridisation (FISH). By culture methods C. jejuni was detectable for only 1 d after spiking whereas bacteria were found from biofilms for at least 1 week after spiking and from outlet water of the reactor for 3 weeks when using FISH. These results suggested that C. jejuni may survive in biofilms and culture methods probably seriously underestimate the real number in water and in biofilms.

The INCA-Pathogens model: An application to the Loimijoki River basin in Finland.

Good hygienic quality of surface waters is essential for drinking water production, irrigation of crops and recreation. Predictions of how and when microbes are transported by rivers are needed to protect downstream water users. In this study we tested the new process-based INCA-Pathogens model in the agricultural Loimijoki River basin (3138km2) in Finland, and we quantified ecosystem services of water purification and water provisioning for drinking and recreation purposes under different scenarios. INCA is a catchment scale process based model to calculate pollutant transfer from terrestrial environment and point sources to the catchment outlet. A clear gradient was observed in the numbers of faecal coliforms along the River Loimijoki. The highest bacterial counts were detected in the middle part of the main stream immediately after small industries and municipal sewage treatment plants. In terms of model performance, the INCA-Pathogen model was able to produce faecal coliform counts and seasonality both in the low pollution level sampling points and in the high pollution level sampling points. The model was sensitive to the parameters defining light decay in river water and in soil compartment, as well as to the amount of faecal coliforms in the manure spread on the fields. The modeling results showed that the number of faecal coliforms repeatedly exceeded 1000 bacteria 100ml-1. Moreover, results lead to the following conclusions: 1) Climate change does not cause a major threat to hygienic water quality as higher precipitation increases runoff and causes diluting effect in the river, 2) Intensification of agriculture is not a threat as long as animal density remains relatively low and environmental legislation is followed, 3) More intensive agriculture without environmental legislation causes a threat especially in tributaries with high field percentage and animal density, and 4) Hygienic water quality in the River Loimijoki can best be improved by improving sewage treatment. We conclude that this catchment scale model is a useful tool for addressing catchment management and water treatment planning issues.

Expression of rat gene for L-gulono-gamma-lactone oxidase, the key enzyme of L-ascorbic acid biosynthesis, in guinea pig cells and in teleost fish rainbow trout (Oncorhynchus mykiss).

The ability of rainbow trout liver and kidney preparations to produce L-ascorbic acid with an added source of L-gulono-gamma-lactone oxidase (GLO) and the absence of their own GLO activity suggested that the reason for the absence of L-ascorbic acid biosynthesis in fish and in guinea pig, a scurvy-prone mammal, can be similar. Nevertheless, results of rat GLO cDNA expression in guinea pig cells and in rainbow trout proved different. In guinea pig cells, rat GLO was expressed in a functional form. Regardless of recombinant GLO transcripts detected in rainbow trout embryos, alevins and in juvenile fish, neither GLO protein nor GLO activity were found. Furthermore, production of L-ascorbic acid in transgenic rainbow trout was not revealed in feeding tests with vitamin C-free diets or after direct administration of L-gulono-gamma-lactone. These results indicate that conditions required for translation or stability of rat GLO are absent in rainbow trout tissues.

Cloning and characterization of glucose transporter in teleost fish rainbow trout (Oncorhynchus mykiss).

The facilitated diffusion of monosaccharides across the plasma membrane is mediated by glucose transporters (GLUTs). In contrast to mammals, the glucose transport system of lower vertebrates remains unexplored. We detected glucose transport activity in rainbow trout embryos. Two GLUTs sharing 83% amino acid identity were cloned from juvenile fish, these have been denoted OnmyGLUT1A and OnmyGLUT1B. In adult trout OnmyGLUT1A is predominantly expressed in the heart with low expression in other tissues. An inverse terminal repeat of a Tc1-like transposable element was found in the 3'-untranslated region of OnmyGLUT1B. Phylogenetic analysis suggested that rainbow trout genes share a common ancestor with higher vertebrate GLUT1. We also found GLUT genes in several salmonid species.

Campylobacter enteritis in 188 hospitalized patients.

The clinical picture and epidemiologic characteristics of infection due to Campylobacter fetus subspecies jejuni were studied in 188 patients hospitalized in Finland during a three-year period. All but two patients had diarrhea; 90% had abdominal pain, fever, and fatigue; half had vomiting and headache; one third experienced electrolyte disturbances; and one fifth of the patients had other complications, most commonly pancreatitis (6%) and arthritis (5%). All age groups were affected, most usually those who were 0 to 9 years old and 20 to 29 years old. The incidence of domestic cases increased during the summer months. With only three exceptions (1.3%), all jejuni strains were sensitive to erythromycin. Among Finns who visited ten popular tourist countries, the incidence of hospitalized C jejuni enteritis cases varied from 0 to 63 per 100,000 travelers.

Ingredients and contaminants of traditional alcoholic beverages in Tanzania.

Home-made but commercially available alcoholic beverages were collected in Dar es Salaam, Tanzania and analysed for their congener alcohol, additive, aflatoxin and heavy metal contents. Ethanol concentrations of the 15 brewed samples ranged from 2.2 to 8.5% w/v whilst the 2 distilled samples contained ethanol 24.2 and 29.3% w/v. Aflatoxin B1 was found in 9 brewed beverages, suggesting the use of contaminated grain or fruit for their production. The amount of zinc in 4 samples was double the World Health Organization recommended maximum for drinking water (5 mg/litre). One brewed beverage contained toxic amount of manganese (12.8 mg/litre). Both distilled spirits were rich in fusel alcohols and one was fortified by caffeine. The results suggested that impurities and contaminants possibly associated with severe health risks, including carcinogens, are often found in traditional alcoholic beverages. Continuous daily drinking of these beverages is certain to increase health risks. Contaminated grain or fruit rejected from foodstuff production should not be used for the production of alcoholic beverages.

Changes in tissue cellularity are associated with growth enhancement in genetically modified arctic char (Salvelinus alpinus L.) carrying recombinant growth hormone gene.

Biochemical and histological analyses were used to study the number and size of cells (cellularity) in tissues of fast-growing, genetically modified Arctic char (Salvelinus alpinus L.), overexpressing sockeye salmon (Oncorhynchus nerka) growth hormone gene (OnGH1). DNA contents of muscle, heart, and liver were compared in transformed, sibling (age control) and 1 year older (size control) char. Total white muscle cross-sectional area, white muscle fiber number, and total nuclei number within the muscle tissue were determined from one complete half-section of each fish. The analyzed tissues responded differently to growth hormone overproduction. In muscle tissue of OnGH1-transformed char, the enhanced growth was clearly associated with proliferation of muscle cells (hyperplasia), whereas in heart tissue both cell proliferation and increase in cell size (hypertrophy) were enhanced. The relative DNA concentration in the liver of transformed char was significantly greater than that of control fish, suggesting reduction in size of hepatic cells.

Monosaccharide uptake in common carp (Cyprinus carpio) EPC cells is mediated by a facilitative glucose carrier.

In most animal cells, transport of monosaccharides across the plasma membrane is mediated by glucose transporters (GLUT). Mammals express at least five distinct transporters (GLUTs 1--5), which are well characterised both functionally and genetically. In contrast, the glucose transport system of fish remains poorly studied. Here we report studies of hexose uptake in carp EPC cells and cloning of a glucose transporter cDNA from these cells. Transport of radio-labelled methylglucose (3-OMG) followed Michaelis--Menten kinetics with a K(m) value (8.5 mM) similar to that of mammalian cells. The inhibition of transport by cytochalasin B and phloretin, but not by phloridzin or cyanide, strongly suggested the existence of a facilitative carrier. D-Glucose, 2-deoxyglucose, 3-OMG, D-mannose and D-xylose were competitive inhibitors of 3-OMG uptake, while L-glucose, mannitol, D-fructose, D-ribose and sucrose did not compete with 3-OMG. We cloned a carp glucose transporter (CyiGLUT1), using RT-PCR and RACE strategies. CyiGLUT1 was different from known carp and zebrafish EST sequences. The complete cDNA (3060 bp) contained one open reading frame encoding a predicted protein of 478 amino acids. The deduced amino acid sequence shared 78% identity with mammalian and avian GLUT1 proteins. Key amino acids involved in substrate selection and catalysis of mammalian GLUTs were conserved in the carp transporter.

A prospective study of the precursors to problem drinking in young adulthood.

This study was part of the Jyväskylä Longitudinal Study on Social Development. The subjects (196 males, 173 females) were studied at age 8, 14 and 26. Three components in drinking habits were obtained at age 26: social, problem and controlled drinking. Moderate to severe problem drinking was obtained for 26% of the men and 1% of the women, and mild problem drinking for 23% of the men and 15% of the women. Problem drinking (defined by the CAGE Questionnaire, arrest for alcohol abuse and other indicators of heavy drinking) was directly accounted for by poor school success at age 14 and, for men, by conduct problems and low anxiety. Variables at age 8 that contributed indirectly to adult problem drinking were aggression, low anxiety, low prosociality and poor school success for men, and high anxiety and poor school success for women. Women and men differed in the effect of social anxiety; in men, anxiety was a protective factor against problem drinking; in women, it was a risk factor. Although conduct problems often precede severe problem drinking, other risk factors may exist among sons of alcoholic parents. Parental drinking had a significant threshold effect on male off-springs' drinking: if parental drinking was low, there was less problem drinking among the male offspring than if a mild dependence on alcohol was observed in the parents.