Contamination by Norovirus and Adenovirus on Environmental Surfaces and in Hands of Conscripts in Two Finnish Garrisons.

This study investigated the presence of norovirus and adenovirus, especially enteric adenovirus, on the environmental surfaces (n = 481) and military conscripts' hands (n = 109) in two Finnish garrisons (A and B) in 2013 and 2014. A questionnaire study was conducted to reveal possible correlations between viral findings on the conscripts' hands and their acute gastroenteritis symptoms. In addition to the swab samples, 14 fecal samples were obtained for viral analysis. In total, norovirus was present in 9.0 % of the surface swabs in 2013, whereas enteric adenovirus was present in 0.0 % and non-enteric adenovirus in 9.4 %. In the same year, 2.6 % of the hand swabs contained norovirus, 2.6 % enteric adenovirus, and 40.3 % non-enteric adenovirus. Norovirus GI.6 was continually detected on the surfaces of garrison A, and identical virus was detected in some of the fecal samples. In garrison B, two slightly different norovirus GII.4 strains were present on the surfaces. The questionnaires revealed no recent acute gastroenteritis cases in garrison A, but in garrison B, where the norovirus-positive hand swabs were collected, 30.6 % of the conscripts reported of recent symptoms. In 2014, norovirus was rarely detected, but adenovirus was again frequently present, both on the surfaces and hands. Taken together, our results suggest that gastroenteritis outbreaks occurred in 2013, but not in 2014. Due to the low number of hand swabs positive for enteric viruses, no conclusions about associations between viral findings and gastroenteritis symptoms could be drawn. This study increased our understanding of the possible transmission of viruses via contaminated environment and hands.

Pilot-scale continuous ultrasonic cleaning equipment reduces Listeria monocytogenes levels on conveyor belts.

Ultrasonic cleaning of a conveyor belt was studied by building a pilot-scale conveyor with an ultrasonic cleaning bath. A piece of the stainless steel conveyor belt was contaminated with meat-based soil and Listeria monocytogenes strains (V1, V3, and B9) and incubated for 72 h to allow bacteria to attach to the conveyor belt surfaces. The effect of ultrasound with a potassium hydroxide-based cleaning detergent was determined by using the cleaning bath at 45 and 50 degrees C for 30 s with and without ultrasound. The detachment of L. monocytogenes from the conveyor belt caused by the ultrasonic treatment was significantly greater at 45 degrees C (independent samples t test, P < 0.001) and at 50 degrees C (independent samples t test, P = 0.04) than without ultrasound. Ultrasonic cleaning efficiency was tested with different cleaning durations (10, 15, 20, and 30 s) and temperatures (30, 45, and 50 degrees C). The differences in the log reduction between cleaning treatments were analyzed by analysis of variance with Tamhane's T2 posthoc test using SPSS (Chicago, IL). The lengthening of the treatment time from 10 to 30 s did not significantly increase the detachment of L. monocytogenes (ANOVA 0.633). At 30 degrees C and at the longest time tested (30 s), the treatment reduced L. monocytogenes counts by only 2.68 log units. However, an increase in temperature from 30 to 50 degrees C improved the effect of the ultrasonic treatment significantly (P < 0.01). Ultrasonic cleaning for 10 s at 50 degrees C reduced L. monocytogenes counts by more than 5 log units. These results indicate that ultrasonic cleaning of a conveyor belt is effective even with short treatment times.

High level of antimicrobial resistance in Campylobacter jejuni isolated from broiler chickens in Estonia in 2005 and 2006.

The development of antimicrobial resistance in Campylobacter jejuni and Campylobacter coli is a matter of increasing concern. Because campylobacteriosis is transmitted to humans usually via food of animal origin, the presence of antimicrobial-resistant campylobacters in broiler chickens has important public health implications. The aim of our study was to analyze resistance patterns of C. jejuni isolated from fecal samples collected at a large Estonian chicken farm, from cecal contents collected at slaughterhouses, and from meat samples collected at the retail establishments in 2005 and 2006. A total of 131 C. jejuni isolates were collected over a 13-month period and tested by the broth microdilution VetMIC method (National Veterinary Institute, Uppsala, Sweden) to determine the MICs of various antimicrobials. Resistance to one or more antimicrobials was detected in 104 (79.4%) of the 131 isolates. High proportions of the isolates were resistant to enrofloxacin (73.3%) and nalidixic acid (75.6%). Multidrug resistance (resistance to three or more unrelated antimicrobials) was detected in 36 isolates (27.5%), all of which were resistant to enrofloxacin. Multidrug resistance was significantly associated with enrofloxacin resistance (P < 0.01), and the use of enrofloxacin may select for multiresistant strains.

Factors associated with Listeria monocytogenes contamination of cold-smoked pork products produced in Latvia and Lithuania.

A total of 312 samples of sliced, vacuum packaged, cold-smoked pork from 15 meat processing plants in Latvia and Lithuania, obtained over a 15-month period from 2003 until 2004, were analyzed for the presence of Listeria monocytogenes at the end of their shelf-life. Overall, 120 samples (38%) tested positive for L. monocytogenes. Despite the long storing period, the levels of L. monocytogenes in cold-smoked pork products were low. Manufacturing processes were studied at seven meat processing plants. A new approach with a logistic multivariable regression model was applied to identify the main factors associated with L. monocytogenes contamination during the manufacturing of cold-smoked pork products. Brining by injection was a significant factor (odds ratio 10.66; P<0.05) for contamination of product with L. monocytogenes. Moreover, long cold-smoking times (> or = 12 h) had a significant predictive value (odds ratio 24.38; P<0.014) for a sample to test positive for L. monocytogenes. Pulsed-field gel electrophoresis results indicated that various sources of L. monocytogenes contamination existed over periods of time in several meat processing plants. In two meat processing plants, persistent L. monocytogenes strains belonging to serotypes 1/2a and 1/2c were found.

Evaluation of the lactose Tergitol-7, m-Endo LES, Colilert 18, Readycult Coliforms 100, Water-Check-100, 3M Petrifilm EC and DryCult Coliform test methods for detection of total coliforms and Escherichia coli in water samples.

In this study we compared the reference membrane filtration (MF) lactose Tergitol-7 (LTTC) method ISO 9308-1:2000 with the MF m-Endo LES method SFS 3016:2001, the defined substrate chromogenic/fluorogenic Colilert 18, Readycult Coliforms and Water Check methods, and ready-made culture media, 3M Petrifilm EC and DryCult Coli methods for the detection of coliforms and Escherichia coli in various water samples. When the results of E. coli detection were compared between test methods, the highest agreement (both tests negative or positive) with the LTTC method was calculated for the m-Endo LES method (83.6%), followed by Colilert 18 (82.7%), Water-Check (81.8%) and Readycult (78.4%), whereas Petrifilm EC (70.6%) and DryCult Coli (68.9%) showed the weakest agreement. The m-Endo LES method was the only method showing no statistical difference in E. coli counts compared with the LTTC method, whereas the Colilert 18 and Readycult methods gave significantly higher counts for E. coli than the LTTC method. In general, those tests based on the analysis of a 1-ml sample (Petrifilm EC and DryCult Coli) showed weak sensitivity (39.5-52.5%) but high specificity (90.9-78.8%).

Contamination routes of Clostridium botulinum in the honey production environment.

Factors influencing Clostridium botulinum contamination in the honey production environment were evaluated in a 3-year survey. A number of 1,168 samples from 100 apiaries and related facilities were analysed for the presence of C. botulinum types A, B, E and F, using multiplex polymerase chain reaction targeted to botA, botB, botE and botF genes. Production methods and environmental factors were registered using a questionnaire and by personal observation. Clostridium botulinum was shown to be a common finding throughout the whole honey production chain, and the type most frequently detected was group I type B. In a pulsed-field gel electrophoresis (PFGE) analysis of 202 group I type B isolates, only six different PFGE profiles were observed, of which two clearly distinct profiles predominated. This may indicate the existence of at least two different genetic lineages. The high prevalence of C. botulinum in soil and in samples associated with beeswax suggests the accumulation of soil-derived botulinal spores in wax. Additionally, according to Spearman's rank order correlation and multivariate analysis, production hygiene-dependent factors have a significant influence on the contamination, and thus the number and frequency of C. botulinum spores in honey could possibly be diminished by increasing hygienic level in honey production.

Monitoring of Cryptosporidium and Giardia in the Vantaa river basin, southern Finland.

We compared two sampling methods to assess the contamination of the Vantaa river basin by Giardia cysts and Cryptosporidium oocysts: 10-1 grab samples, the common river mussel Anadonta piscinalis, were analysed for concentration of (oo)cysts from river water. The samples were collected 2-5 times in autumn 2001 from four wastewater treatment plants and four river water sites located downstream of the plants, and six times from raw water of a drinking water plant using the river as water source. The presence of Giardia and Cryptosporidium was analysed by IF microscopy and PCR. Both cysts and oocysts were detected at all sampling sites, but oocysts were more common than cysts in river water samples. In contrast, cysts were more common in A. piscinalis. Most Cryptosporidium-positive samples were of genotype 2 and Giardia were assemblage B. In river water, MPN of Escherichia coli did not correlate to the presence of (oo)cysts. In conclusion, low (oo)cyst counts were regularly identified in the Vantaa river basin which is contaminated by discharges of treated wastewater of human origin. In general, both methods to appropriate to detect (oo)cysts, but grab samples yielded more positive results. Grab sampling is also more practical and less expensive than analysis of A. piscinalis.

Elimination of botulinum neurotoxin (BoNT) type B from drinking water by small-scale (personal-use) water purification devices and detection of BoNT in water samples.

Seven small-scale drinking water purification devices were evaluated for their capacity to eliminate botulinum neurotoxin (BoNT) type B from drinking water. Influent water inoculated with toxic Clostridium botulinum cultures and effluent purified water samples were tested for the presence of BoNT by using a standard mouse bioassay and two commercial rapid enzyme immunoassays (EIAs). The water purification devices based on filtration through ceramic or membrane filters with a pore size of 0.2 to 0.4 microm or irradiation from a low-pressure UV-lamp (254 nm) failed to remove BoNT from raw water (reduction of < 0.1 log10 units). A single device based on reverse osmosis was capable of removing the BoNT to a level below the detection limit of the mouse bioassay (reduction of > 2.3 log10 units). The rapid EIAs intended for the detection of BoNT from various types of samples failed to detect BoNT from aqueous samples containing an estimated concentration of BoNT of 396,000 ng/liter.

Meta-analysis in assessment of the prevalence and annual incidence of Giardia spp. and Cryptosporidium spp. infections in humans in the Nordic countries.

We aimed to apply the meta-analysis in the studies of protozoan pathogens in order to obtain a general overview of the prevalence and annual incidence of Giardia spp. and Cryptosporidium spp. infections in asymptomatic and symptomatic human populations in the Nordic countries of Denmark, Finland, Norway and Sweden. In combining the data of 13 clinically and methodologically non-heterogeneous studies published before 2004 using the random effects model with DerSimonian-Laird estimator, we estimated the prevalence (% prevalence: 95% confidence limits) of Giardia cases in the asymptomatic (i.e. no gastroenteric symptoms) general population to be 2.97% (2.64; 3.31) and in the symptomatic population 5.81% (5.34; 6.30). For Cryptosporidium the prevalences were 0.99% (0.81; 1.19) and 2.91% (2.71; 3.12), respectively. In analyzing the data, we estimated that there will be 4670 (4300; 5060) symptomatic cases of Giardia and 3340 (3110; 3580) symptomatic cases of Cryptosporidium annually per 100,000 general population in the Nordic countries. The vast majority of cases will remain unregistered in the national registers of infectious diseases, since for single registered cases there will be 254-867 cases of Giardia undetected/unregistered and 4072 to 15,181 cases of Cryptosporidium, respectively.

Campylobacter spp., Giardia spp., Cryptosporidium spp., noroviruses, and indicator organisms in surface water in southwestern Finland, 2000-2001.

A total of 139 surface water samples from seven lakes and 15 rivers in southwestern Finland were analyzed during five consecutive seasons from autumn 2000 to autumn 2001 for the presence of various enteropathogens (Campylobacter spp., Giardia spp., Cryptosporidium spp., and noroviruses) and fecal indicators (thermotolerant coliforms, Escherichia coli, Clostridium perfringens, and F-RNA bacteriophages) and for physicochemical parameters (turbidity and temperature); this was the first such systematic study. Altogether, 41.0% (57 of 139) of the samples were positive for at least one of the pathogens; 17.3% were positive for Campylobacter spp. (45.8% of the positive samples contained Campylobacter jejuni, 25.0% contained Campylobacter lari, 4.2% contained Campylobacter coli, and 25.0% contained Campylobacter isolates that were not identified), 13.7% were positive for Giardia spp., 10.1% were positive for Cryptosporidium spp., and 9.4% were positive for noroviruses (23.0% of the positive samples contained genogroup I and 77.0% contained genogroup II). The samples were positive for enteropathogens significantly (P < 0.05) less frequently during the winter season than during the other sampling seasons. No significant differences in the prevalence of enteropathogens were found when rivers and lakes were compared. The presence of thermotolerant coliforms, E. coli, and C. perfringens had significant bivariate nonparametric Spearman's rank order correlation coefficients (P < 0.001) with samples that were positive for one or more of the pathogens analyzed. The absence of these indicators in a logistic regression model was found to have significant predictive value (odds ratios, 1.15 x 10(8), 7.57, and 2.74, respectively; P < 0.05) for a sample that was negative for the pathogens analyzed. There were no significant correlations between counts or count levels for thermotolerant coliforms or E. coli or the presence of F-RNA phages and pathogens in the samples analyzed.

An IC-PCR method for detection of Cryptosporidium and Giardia in natural surface waters in Finland.

We developed an immunocapture-based polymerase chain reaction (PCR) assay for simultaneous detection of Cryptosporidium parvum oocysts and Giardia intestinalis cysts in surface water. Using primer pairs Cry9/Cry15 and LaxA/LaxB for Cryptosporidium and Gdh1/Gdh4 for Giardia, the sensitivity of the entire detection procedure (dealing with concentration, separation, DNA purification and PCR amplification) was at the level of 50-100 oocysts and cysts. Of 54 surface water samples, 4 were positive for Cryptosporidium and 1 for Giardia. Cryptosporidium and Giardia were detected for the first time in surface water in Finland.