Evaluation of Criminal Sanctions Concerning Violations of Cattle and Pig Welfare.

EU legislation requires the violations of animal welfare standards to be sanctioned. Our aim was to evaluate criminal sanctions concerning violations of cattle and pig welfare on Finnish farms. We analyzed 196 court cases heard in Finnish district courts from 2011 to 2016. Almost all the cases (95%) concerned the violations of cattle welfare, of which 61% occurred on small farms. The lack of cleanliness and inadequate feeding and watering were the most common reported violations. Median time span from the start date of an offending to a judgement was slightly less than two years. Of the cases, 96% resulted in conviction. The court did not perceive the violations as being highly blameworthy as a small fine and a short conditional imprisonment were the most often imposed sanctions. A ban on the keeping of animals was used as a precautionary measure in half of the cases. Veterinarians were shown to have an important role in the initiation of criminal procedures, providing evidence for the police, and acting as witnesses. Therefore, it is crucial to achieve a well-functioning collaboration between veterinarians and the police and prosecutors. The expertise of these authorities on animal welfare legislation should also be emphasized to improve the efficacy of criminal procedures.

Veterinarians Experience Animal Welfare Control Work as Stressful.

The aim of the study was to evaluate the job satisfaction of official veterinarians working in the field of animal welfare control and identify both positive features and challenges of their work. An electronic questionnaire was designed to evaluate job satisfaction. The questionnaire was responded to by 73 of the 98 Finnish official veterinarians working in the field of animal welfare control. The Spearman's rank correlation coefficient was used to evaluate the relation between stress and different work-related factors. More than half of the respondents reported work-related stress or fatigue. Threatening situations, disturbed work-private life balance and a high amount of overtime work were found to be frequent underlying causes of stress. Fieldwork, especially when working alone, was perceived as the most challenging part of the work. Of the respondents, three out of four performed animal welfare inspections mainly alone. Although the respondents reported getting additional help to perform an inspection most of the times they needed it, a wish to work in a pair was highlighted. The results of the present study indicate that official veterinarians often experience work-related stress and fatigue. By testing interventions shown to be beneficial, such as providing adequate support within the work community, decreasing the workload and enabling inspections to be done in pairs, job satisfaction could be improved.

Assessment of Welfare Problems in Finnish Cattle and Pig Farms Based on Official Inspection Reports.

The competent authorities of the Member States of the European Union are required to perform animal welfare inspections on livestock farms. The data obtained from these official inspections performed in Finnish cattle and pig farms in 2010-2015 were used with the aim of estimating the prevalence of the most common non-compliances and identifying underlying risk factors. The prevalence of non-compliant cattle and pig farms was 24.2% and 27.9%, respectively. In cattle, the most common problem was an inadequate lying area followed by deficient housing conditions for calves; in pigs, it was a lack of enrichment material. The non-compliances concerning cattle were most frequently detected in autumn and in farms with small herd size, with tie-stall housing and outdoor rearing year-round. The pig farms with a farrow-to-finish unit had a higher prevalence of non-compliances than other production types. The prevalence of the non-compliant farms differed notably between the regions. It can be concluded that the cattle welfare inspections should be performed with a focus on the cold and rainy seasons and at small farms, whereas the pig welfare inspections should mainly focus on farrow-to-finish units. The data received from official inspections should be efficiently utilized in the development of animal welfare inspection system, with the aim of risk-based, consistent and uniform inspections. In addition, the data should be utilized in targeting information for farmers.

Towards more consistent and effective food control: learning from the views of food business operators.

We surveyed the opinions of Finnish food business operators (FBOs) about the uniformity of local official food control and its importance for dairy, fishery and meat plants. A total of 136 FBOs responded to the questionnaire. Most FBOs considered official food control to be important for food safety and were generally satisfied with its quality. However, they often did not perceive official food control as being uniform, and 23% even considered it arbitrary. Small-sized FBOs were particularly critical of the relevance of control actions. The better the FBOs assessed their cooperation with the inspector, the higher they assessed the quality, uniformity and benefits of official food control. The cooperative approach in control practices should be emphasized to support the positive views of FBOs about official food control, thus promoting food safety. Cross-audits among local food control units are recommended to improve the FBOs' experience of uniformity of food control.

[Food control--the mainstay of food safety]. / Elintarvikevalvonta--elintarviketurvallisuuden tukipilari.

The main responsibility of the local food control lies in the local food and environmental health units. The producer is responsible for the safety of the food produced, and applies an in-house control system to achieve this goal. The local food safety authority evaluates the in-house control system and assures that it is targeted to the most critical steps in the production to efficiently minimize the food safety risks. The main challenges in implementing efficient food control system are the variability in the knowledge of the producer about food hygiene, inadequate resources in control units and harmonization of the control measures in national level.

Comparative genomic hybridization analysis of two predominant Nordic group I (proteolytic) Clostridium botulinum type B clusters.

Comparative genomic hybridization analysis of 32 Nordic group I Clostridium botulinum type B strains isolated from various sources revealed two homogeneous clusters, clusters BI and BII. The type B strains differed from reference strain ATCC 3502 by 413 coding sequence (CDS) probes, sharing 88% of all the ATCC 3502 genes represented on the microarray. The two Nordic type B clusters differed from each other by their response to 145 CDS probes related mainly to transport and binding, adaptive mechanisms, fatty acid biosynthesis, the cell membranes, bacteriophages, and transposon-related elements. The most prominent differences between the two clusters were related to resistance to toxic compounds frequently found in the environment, such as arsenic and cadmium, reflecting different adaptive responses in the evolution of the two clusters. Other relatively variable CDS groups were related to surface structures and the gram-positive cell wall, suggesting that the two clusters possess different antigenic properties. All the type B strains carried CDSs putatively related to capsule formation, which may play a role in adaptation to different environmental and clinical niches. Sequencing showed that representative strains of the two type B clusters both carried subtype B2 neurotoxin genes. As many of the type B strains studied have been isolated from foods or associated with botulism, it is expected that the two group I C. botulinum type B clusters present a public health hazard in Nordic countries. Knowing the genetic and physiological markers of these clusters will assist in targeting control measures against these pathogens.

The detection and prevalence of Clostridium botulinum in pig intestinal samples.

To determine the prevalence of Clostridium botulinum types A, B, E, and/or F in pig intestinal samples, different PCR-based methods were compared using artificially contaminated faeces. The methods included a multiplex PCR targeted to botulinum neurotoxin genes A, B, E, and F or a nested PCR targeted to toxin genes B, E, and F, combined with different pre-enrichment protocols and forms of templates. A method including the two-step enrichment followed by DNA extraction and multiplex PCR yielded the highest number of positives. This assay protocol was employed to investigate 100 pig intestinal samples. The sample materials studied included colon wall, intestinal content, and mucus peeled from the colon wall. Three pigs (3%) were positive for C. botulinum type B, and no other toxinotypes were detected in any sample. The number of positive samples was higher when colon wall or peeled mucus was analyzed compared to the intestinal content. C. botulinum was isolated from two PCR-positive samples and confirmed to be type B by PCR. Both isolates were shown to be proteolytic and thus to represent C. botulinum group I.

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.

Prevalence and diversity of Clostridium botulinum types A, B, E and F in honey produced in the Nordic countries.

A total of 294 honey samples produced in Denmark, Norway and Sweden were studied for the presence of Clostridium botulinum types A, B, E and F by using a multiplex-PCR method. The samples consisted of honeycombs taken directly from beehives, and extracted honey representing several hives or apiaries. The prevalence of C. botulinum showed a significant variation between Denmark, Norway and Sweden, the proportions of positive samples being 26%, 10% and 2%, respectively. The major serotype detected was type B. When analysed with pulsed-field gel electrophoresis (PFGE) using restriction enzyme SacII, the 24 strains isolated produced eight different PFGE patterns. At a similarity level of 95%, four clusters were produced, three of which contained 20 of the 24 analysed strains. One of the clusters included isolates from both Denmark and Norway.

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.

Efficient DNA fingerprinting of Clostridium botulinum types A, B, E, and F by amplified fragment length polymorphism analysis.

Amplified fragment length polymorphism (AFLP) analysis was applied to characterize 33 group I and 37 group II Clostridium botulinum strains. Four restriction enzyme and 30 primer combinations were screened to tailor the AFLP technique for optimal characterization of C. botulinum. The enzyme combination HindIII and HpyCH4IV, with primers having one selective nucleotide apiece (Hind-C and Hpy-A), was selected. AFLP clearly differentiated between C. botulinum groups I and II; group-specific clusters showed <10% similarity between proteolytic and nonproteolytic C. botulinum strains. In addition, group-specific fragments were detected in both groups. All strains studied were typeable by AFLP, and a total of 42 AFLP types were identified. Extensive diversity was observed among strains of C. botulinum type E, whereas group I had lower genetic biodiversity. These results indicate that AFLP is a fast, highly discriminating, and reproducible DNA fingerprinting method with excellent typeability, which, in addition to its suitability for typing at strain level, can be used for C. botulinum group identification.

Diversity of proteolytic Clostridium botulinum strains, determined by a pulsed-field gel electrophoresis approach.

Pulsed-field gel electrophoresis (PFGE) was applied to the study of the similarity of 55 strains of proteolytic Clostridium botulinum (C. botulinum group I) types A, AB, B, and F. Rare-cutting restriction enzymes ApaI, AscI, MluI, NruI, PmeI, RsrII, SacII, SmaI, and XhoI were tested for their suitability for the cleavage of DNA of five proteolytic C. botulinum strains. Of these enzymes, SacII, followed by SmaI and XhoI, produced the most convenient number of fragments for genetic typing and were selected for analysis of the 55 strains. The proteolytic C. botulinum species was found to be heterogeneous. In the majority of cases, PFGE enabled discrimination between individual strains of proteolytic C. botulinum types A and B. The different toxin types were discriminated at an 86% similarity level with both SacII and SmaI and at an 83% similarity level with XhoI. Despite the high heterogeneity, three clusters at a 95% similarity level consisting of more than three strains of different origin were noted. The strains of types A and B showed higher diversity than the type F organisms which formed a single cluster. According to this survey, PFGE is to be considered a useful tool for molecular epidemiological analysis of proteolytic C. botulinum types A and B. However, epidemiological conclusions based on PFGE data only should be made with discretion, since highly similar PFGE patterns were noticed, especially within the type B strains.

Infant botulism acquired from household dust presenting as sudden infant death syndrome.

Clostridium botulinum type B was detected by multiplex PCR in the intestinal contents of a suddenly deceased 11-week-old infant and in vacuum cleaner dust from the patient's household. C. botulinum was also isolated from the deceased infant's intestinal contents and from the household dust. The genetic similarity of the two isolates was demonstrated by pulsed-field gel electrophoresis and randomly amplified polymorphic DNA analysis, thereby confirming that dust may act as a vehicle for infant botulism that results in sudden death.

Type C botulism due to toxic feed affecting 52,000 farmed foxes and minks in Finland.

The largest reported outbreak of type C botulism in fur production animals is described. Epidemiological investigation of 117 out of 157 (response rate, 74.5%) farms revealed that 44,130 animals died or were euthanized, while 8,033 animals with milder symptoms recovered. The overall death rate in all animals at risk was 21.7%. The death rates were significantly higher in blue and shadow foxes (24.2 and 27.8%, respectively) than in silver and blue silver foxes and minks (below 4%). All minks had been immunized against botulinum toxin type C. Deaths were associated with feed manufactured by a local processor, 83 of whose customer farms (70.9%) reported dead or sick animals. Five feedlots out of 19 delivered to the farms on the day preceding the onset of the outbreak (day 2) were associated with a death rate higher than 40%. These feedlots consisted of fresh feed processed on day 2 and feed processed 1 day earlier (day 1). In laboratory analysis, the day 2 feed contained botulinum toxin type C (>600 minimum lethal doses/g), while the day 1 feed did not contain toxin. Toxin was not detected in feed raw-material samples. Clostridium botulinum type C was detected by PCR in some feed components and in feed. However, as the feed temperature was continuously 8 degrees C or below and the pH was continuously 5.6 or below according to the manufacturer, it seems unlikely that spore germination and toxin formation occurred during overnight storage. Hence, the events leading to toxin formation were not determined.

Antibacterial efficiency of Finnish spice essential oils against pathogenic and spoilage bacteria.

The antibacterial properties of 13 essential oils, derived from spices grown in Finland, were examined with an agar diffusion method against 12 bacterial strains. The organisms tested included both spoilage and pathogenic bacteria. The gram-positive bacteria appeared to be more sensitive than the gram-negative organisms, Clostridium botulinum and Clostridium perfringens being the most sensitive. Oregano, savory, and thyme showed the broadest antibacterial activity by distinctly inhibiting the growth of all the organisms tested. By gas chromatography-mass spectrometry analysis, differences were noted in the composition of oregano and thyme oils in comparison to previous reports.

Proteolytic Clostridium botulinum type B in the gastric content of a patient with type E botulism due to whitefish eggs.

Whitefish eggs were confirmed by pulsed-field gel electrophoresis to cause type E foodborne botulism in a 54-year-old patient in Finland. Botulinum neurotoxin and/or nonproteolytic Clostridium botulinum type E organisms were detected in fecal and gastric samples from the patient and in suspected whitefish eggs. Apart from C. botulinum type E, proteolytic type B organisms were detected in the patient's gastric content. This was considered to be insignificant with respect to the clinical disease, suggesting botulinal spores to be occasionally present in the human gastrointestinal tract without any apparent clinical significance. This is the first domestic case of foodborne botulism in Finland.

Thermal inactivation of nonproteolytic Clostridium botulinum type E spores in model fish media and in vacuum-packaged hot-smoked fish products.

Thermal inactivation of nonproteolytic Clostridium botulinum type E spores was investigated in rainbow trout and whitefish media at 75 to 93 degrees C. Lysozyme was applied in the recovery of spores, yielding biphasic thermal destruction curves. Approximately 0.1% of the spores were permeable to lysozyme, showing an increased measured heat resistance. Decimal reduction times for the heat-resistant spore fraction in rainbow trout medium were 255, 98, and 4.2 min at 75, 85, and 93 degrees C, respectively, and those in whitefish medium were 55 and 7.1 min at 81 and 90 degrees C, respectively. The z values were 10.4 degrees C in trout medium and 10.1 degrees C in whitefish medium. Commercial hot-smoking processes employed in five Finnish fish-smoking companies provided reduction in the numbers of spores of nonproteolytic C. botulinum of less than 10(3). An inoculated-pack study revealed that a time-temperature combination of 42 min at 85 degrees C (fish surface temperature) with >70% relative humidity (RH) prevented growth from 10(6) spores in vacuum-packaged hot-smoked rainbow trout fillets and whole whitefish stored for 5 weeks at 8 degrees C. In Finland it is recommended that hot-smoked fish be stored at or below 3 degrees C, further extending product safety. However, heating whitefish for 44 min at 85 degrees C with 10% RH resulted in growth and toxicity in 5 weeks at 8 degrees C. Moist heat thus enhanced spore thermal inactivation and is essential to an effective process. The sensory qualities of safely processed and more lightly processed whitefish were similar, while differences between the sensory qualities of safely processed and lightly processed rainbow trout were observed.

High prevalence of Clostridium botulinum types A and B in honey samples detected by polymerase chain reaction.

A test protocol for reliable detection of Clostridium botulinum types A and B spores in honey by polymerase chain reaction (PCR) was developed and used for a prevalence survey of C. botulinum spores in 190 honey samples. The inhibiting effects of honey on microbial growth and PCR analysis were overcome by using a method of supernatant filtration (SF) in the preparation of the samples before enrichment and PCR. By using this method, an inoculum of 0.1 spore of C. botulinum/g honey could be detected. In the prevalence survey, spores of C. botulinum were detected in 8 (7%) of the 114 Finnish and in 12 (16%) of the 76 imported honey samples. The quantity of spores in PCR-positive samples varied from less than 18 to 140 spores/kg. Neurotoxin gene sequences corresponding to C. botulinum type A were detected in 17 samples and proteolytic type B in 12 samples by PCR analysis. Both types A and B were detected in nine samples. Strains of C. botulinum type A were isolated from 14 and type B from 2 of the 20 PCR-positive samples. This is the first report of type A spores of C. botulinum being detected and isolated in Fennoscandia.