Ari Hörman

Campylobacter spp., Giardia spp., Cryptosporidium spp., Noroviruses, and Indicator Organisms in Surface Water in Southwestern Finland, 2000-2001

ABSTRACT 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 Spearmans 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 × 108, 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.

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.

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 Tamhanes 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.

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

ABSTRACT 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 μm 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.