The Q fever epidemic in The Netherlands: history, onset, response and reflection.

The 2007-2009 human Q fever epidemic in The Netherlands attracted attention due to its magnitude and duration. The current epidemic and the historical background of Q fever in The Netherlands are reviewed according to national and international publications. Seroprevalence studies suggest that Q fever was endemic in The Netherlands several decades before the disease was diagnosed in dairy goats and dairy sheep. This was in 2005 and the increase in humans started in 2007. Q fever abortions were registered on 30 dairy goat and dairy sheep farms between 2005 and 2009. A total of 3523 human cases were notified between 2007 and 2009. Proximity to aborting small ruminants and high numbers of susceptible humans are probably the main causes of the human Q fever outbreak in The Netherlands. In general good monitoring and surveillance systems are necessary to assess the real magnitude of Q fever.

Species identification in meat products using real-time PCR.

One of the most convenient methods for the identification of animal species in processed meat products is the examination of DNA sequences. Real-time polymerase chain reaction (qPCR) techniques are particularly suitable because even small fragments of DNA formed during heat processing of the meat can be amplified and identified. A real-time PCR method has been developed and evaluated for the identification of processed meat products. In test mixtures containing beef, pork, horse, mutton, chicken and turkey, it was possible to identify these species down to a level of 0.05%. By adjusting the number of cycles, it was possible to detect levels as low as 0.01% of these species. Cross-reactivity between these species was not found, except for pure horsemeat (250 ng DNA) in the assay for turkey meat. Cross-reactivity of deer, roe, ostrich, kangaroo, goat, domestic duck, mallard, goose, pigeon, guinea fowl, quail and pheasant was also investigated and it was found that amounts as high as 250 ng DNA of these species in the reaction vial did not result in (false) positive signals except for amounts higher than 125 ng deer DNA and higher than 50 ng pigeon DNA in the determination of chicken and beef, respectively. More than 150 meat samples were examined using DNA hybridization and real-time PCR. A comparison of the results showed a better performance of the real-time procedure compared to DNA hybridization.

Public farms: hygiene and zoonotic agents.

In three successive years, we visited petting farms (n=132), care farms (n=91), and farmyard campsites (n=84), respectively, and completed a standard questionnaire with the objective of determining the hygienic status of these farms and describing hygiene measures implemented to reduce the risk of transmission of zoonotic agents from the animals to humans. For at least 85% of the farms, the overall impression of hygiene was recorded as good. However, more attention must be paid to: informing visitors on hygiene and handwashing, provision of handwashing facilities, and a footwear cleaning facility. Examination of samples of freshly voided faeces resulted in the detection of Shiga toxin-producing Escherichia coli O157 and/or Salmonella spp. and/or Campylobacter spp. at almost two-thirds (64.9%) of the petting farms, and around half of the care farms (56.0%) and farmyard campsites (45.2%). These data reinforce the need for control measures for both public and private farms to reduce human exposure to livestock faeces and thus the risk of transmission of zoonotic diseases. Public awareness of the risk associated with handling animals or faecal material should be increased.

Reduction of campylobacter infections in broiler flocks by application of hygiene measures.

Transmission routes of Campylobacter spp. in broilers and possibilities for prevention of infections were studied on two Dutch broiler farms. The occurrence of Campylobacter spp. was studied in successive broiler flocks, in the environment of the farms and in some of the parent flocks involved. Isolates of Campylobacter spp. were typed by using randomly amplified polymorphic DNA (RAPD) analysis. The results indicate that broiler flocks become infected from environmental sources. The typing results suggest that on one farm transmission of Campylobacter spp. occurred from cattle to broilers via the farmer's footwear. After several campylobacter positive broiler cycles hygiene measures, including thorough cleaning and disinfection procedures, change of footwear at the entrance of each broiler house, control of vermin and other hygienic precautions, were introduced on both farms in order to prevent transmission of Campylobacter spp. from the farm environment to the broilers. The results indicate that the application of hygiene measures significantly reduced campylobacter infections of broiler flocks on both farms.

Epidemiological study on risk factors and risk reducing measures for campylobacter infections in Dutch broiler flocks.

From September 1991 until August 1993 an epidemiological study involving 20 Dutch broiler farms was conducted to identify risk factors and risk reducing measures for campylobacter infections in broiler flocks. Campylobacter spp. were detected in 64 (57%) of the 112 broiler flocks and in 25 (63%) of the 40 broiler cycles examined. Univariate analysis of farm management data was performed followed by logistic regression analysis of selected risk and risk reducing factors. The presence of other farm animals, including pigs, cattle, sheep and fowl, other than broilers, was found to be independently associated with an increased risk of campylobacter infections in broiler flocks (odds ratio (OR) = 11.81; P = 0.041). Further, the results indicate that application of specific hygiene measures during the rearing period, such as washing hands before tending the broiler flocks, the use of separate boots for each broiler house and the use of footbath disinfection when entering a broiler house, may significantly reduce the risk of campylobacter infections in broiler flocks.

A selective medium for the isolation of Arcobacter from meats.

A method, including enrichment in Arcobacter Selective Broth (ASB) and isolation on semisolid Arcobacter Selective Medium (ASM) under aerobic conditions at 24 degrees C, is described for the isolation of Arcobacter from retail meat products. Selective agents used in ASB and ASM were cefoperazone, trimethoprim, piperacillin and cycloheximide. Arcobacters were isolated from 53 (24.1%) of 220 poultry meat products and also, at lower incidence from samples of beef and pork. The isolates were identified as A. butzleri or A. butzleri-like and belonged to a wide variety of serotypes and biotypes.