Biomedical subjects
Publications and source records attributed to H Kruse.
In this study comprising isolates from 2001 to 2003, resistance was considerably more widespread among Campylobacter jejuni from humans infected abroad than infected within Norway. The discrepancy was particularly notable for fluoroquinolone resistance (67.4% vs. 6.5%). This is probably a reflection of a low resistance prevalence in Norwegian broiler isolates (1.2% fluoroquinolone resistant).
We examined the occurrence and diversity of Campylobacter jejuni on broiler carcasses during slaughter of an infected flock and in the slaughterhouse environment during slaughter and postdisinfection before a new production run. During the slaughter of a known C. jejuni infected broiler flock, samples were taken from broiler carcasses at 7 different stages during the process. Thirty-seven sites in the slaughterhouse environment were sampled both during process and postdisinfection. The samples were analyzed for C. jejuni, and genetic fingerprinting was performed using amplified fragment length polymorphism. All carcass samples were positive. Of the environmental samples collected during slaughter, 89% were positive; 100% of those from the arrival, stunning, scalding, defeathering, and evisceration facilities and 67% of those from the cooling and sorting facilities. Postdisinfection, 41% of the samples were positive; 71% of those from the arrival and stunning area, 60% of those from the scalding and defeathering area, and 20% of those from the evisceration, cooling, and sorting area. The C. jejuni isolates (n = 60) recovered were grouped into 4 different amplified fragment length polymorphism clones with a similarity index of 95% or greater. All isolates obtained from the flock and 94% of the isolates obtained from the environment during slaughtering belonged to clone A, whereas 1 environmental isolate belonged to each of the clones B and C. Isolates from clones A, B, and D were present postdisinfection. Only clone B was detected on flocks slaughtered during the previous week. The high level and continuous presence of Campylobacter in the environment constitutes a risk for transmission to negative carcasses. In Norway, where above 96% of the broiler flocks are Campylobacter-negative, this aspect is of special importance. The ability of Campylobacter to remain in the slaughterhouse environment through washing and disinfection is associated with constructional conditions of equipment and buildings, complicating cleaning and providing sufficient moisture. To reduce the probability of the workers acquiring campylobacteriosis, precautions should be taken when slaughtering Campylobacter-positive flocks.
AIMS: To investigate the genetic diversity of Campylobacter in broilers and in the environment of broiler farms, to compare the genetic profiles and describe critical factors for transmission to broilers. METHODS AND RESULTS: Flocks at three of four investigated farms became colonized with Campylobacter. The total proportion of Campylobacter-positive samples at different farms varied from 20% to 42%. The farm with the poorest biosecurity routines had broilers that became infected earliest, the highest proportion of positive samples and the highest genetic diversity among the broiler Campylobacter isolates. Campylobacter isolates within common amplified-fragment length polymorphism (AFLP) clusters (95-100%) were found to be present in outdoor environment and in broilers at adjacent farms before they were found in the broilers. A large presence of Campylobacter in the farm environment was demonstrated after the broilers were infected. A high genetic diversity was found among Campylobacter present in the outdoor environment, where certain Campylobacter clusters were found for periods of up to 6 weeks. CONCLUSION: Confirmation by AFLP indicates adjacent poultry farms and outdoor environment as major sources of Campylobacter infection of broilers, this being the novel achievements. SIGNIFICANCE AND IMPACT OF THE STUDY: The results provide more exact knowledge on transmission of Campylobacter at farm level, helpful for developing optimal preventive strategies.
Environmental reservoirs of glycopeptide-resistant enterococci (GRE) in Norway have been linked to former growth promoting use of the glycopeptide avoparcin in poultry production. We have examined the prevalence of fecal GRE in poultry and poultry farmers 3 to 8 years after the Norwegian avoparcin ban in 1995 and performed molecular analyses of the GRE population. Fecal samples from poultry farmers and their flocks on 29 previously avoparcin-exposed farms were collected on five occasions during the study period (1998 to 2003). All flocks (100%) were GRE positive in 1998. Throughout the study period, 78.5% of the poultry samples were GRE positive. Glycopeptide-resistant Enterococcus faecium (GREF) was isolated from 27.6% of the farmer samples in 1998 and from 27.8% of the samples collected between 1998 and 2003. The prevalence of fecal GRE in poultry declined significantly during the study period, but prevalence in samples from the farmers did not decline. PCR analysis revealed a specific Tn1546-plasmid junction fragment in 93.9% of E. faecium isolates. A putative postsegregation killing (PSK) system linked to Tn1546 was detected in 97.1% of the isolates examined. Multilocus sequence typing of glycopeptide-susceptible (n = 10) and -resistant (n = 10) E. faecium isolates from humans (n = 10) and poultry (n = 10) on two farms displayed 17 different sequence types. The study confirms the continuing persistence of a widespread common plasmid-mediated vanA-pRE25-PSK element within a heterogeneous GRE population on Norwegian poultry farms 8 years after the avoparcin ban. Moreover, it suggests an important role of PSK systems in the maintenance of antimicrobial resistance determinants in reservoirs without apparent antimicrobial selection.
The evolutionary processes responsible for the long-term persistence of glycopeptide-resistant Enterococcus faecium (GREF) in nonselective environments were addressed by genetic analyses of E. faecium populations in animals and humans on two Norwegian poultry farms that were previously exposed to avoparcin. A total of 222 fecal GREF (n = 136) and glycopeptide-susceptible (n = 86) E. faecium (GSEF) isolates were obtained from farmers and poultry on three separate occasions in 1998 and 1999. Pulsed-field gel electrophoresis (PFGE) and plasmid DNA analyses discerned 22 GREF and 32 GSEF PFGE types within shifting polyclonal animal and human E. faecium populations and indicated the presence of transferable plasmid-mediated vanA resistance, respectively. Examples of dominant, persistent GREF PFGE types supported the notion that environmentally well-adapted GREF types may counteract the reversal of resistance. PFGE analyses, sequencing of the purK housekeeping gene, and partial typing of vanA-containing Tn1546 suggested a common animal and human reservoir of glycopeptide resistance. Inverse PCR amplification and sequence analyses targeting the right end of the Tn1546-plasmid junction fragment strongly indicated the presence of a common single Tn1546-plasmid-mediated element in 20 of 22 GREF PFGE types. This observation was further strengthened by vanY-vanZ hybridization analyses of plasmid DNAs as well as the finding of a physical linkage between Tn1546 and a putative postsegregation killing system for seven GREF PFGE types. In conclusion, our observations suggest that the molecular unit of persistence of glycopeptide resistance is a common mobile plasmid-mediated vanA-containing element within a polyclonal GREF population that changes over time. In addition, we propose that "plasmid addiction systems" may contribute to the persistence of GREF in nonselective environments.
Rectal swabs from healthy cats and dogs, and from dogs and cats with clinical diarrhoea were collected approximately every third month from May 2000 to June 2001 from six small-animal practices throughout Norway. A questionnaire was filled in for each animal. Of the 301 healthy cats sampled, 54 (18%) were positive for Campylobacter, compared to 5 out of 31 (16%) cats with diarrhoea. Campylobacter jejuni was isolated from 11 (3%), C. upsaliensis from 42 (13%) and C. coli from 2 (0.6%) of the cats sampled. Isolates from four cats (1%) could not be specified. Of the 529 healthy dogs, 124 (23%) were positive for Campylobacter, compared to 18 of 66 (27%) dogs with diarrhoea. C. jejuni was isolated from 20 (3%) and C. upsaliensis from 117 (20%) of the dogs sampled. Isolates from five dogs (0.8%) could not be specified. Eighteen out of the 20 investigated C. upsaliensis samples were resistant to streptomycin. The clinically healthy animals were included in the analysis to identify factors associated with Campylobacter prevalence. The cat model had low classification ability. The dog-data model indicated increased odds of infection with Campylobacter for dogs </=1 year, and in dogs sampled during the spring. No difference was observed between the prevalence of Campylobacter infections in cats and dogs with diarrhoea and healthy animals.
Commercial starter culture bacteria are widely used in the production of dairy products and could represent a potential source for spread of genes encoding resistance to antimicrobial agents. To learn more about the antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products, a total of 189 isolates of lactic acid bacteria were examined for susceptibility to ampicillin, penicillin G, cephalothin, vancomycin, bacitracin, gentamicin, streptomycin, erythromycin, tetracycline, chloramphenicol, quinupristin/dalfopristin, ciprofloxacin, trimethoprim and sulphadiazine using Etest for MIC determination. Most of the isolates (140) originated from 39 dairy products (yoghurt, sour cream, fermented milk and cheese), while 49 were isolated directly from nine commercial cultures. The bacteria belonged to the genera Lactobacillus, Lactococcus, Leuconostoc and Streptococcus. Only one of the 189 isolates was classified as resistant to an antimicrobial agent included in the study. This isolate, a lactobacillus, was classified as high level resistant to streptomycin. The remaining isolates were not classified as resistant to the antimicrobial agents included other than to those they are known to have a natural reduced susceptibility to. Thus, starter culture bacteria in Norwegian dairy products do not seem to represent a source for spread of genes encoding resistance to antimicrobial agents.
Avoparcin was used as a growth promoting feed additive in Norwegian broiler and turkey production from 1986 until it was banned in 1995, when an association between vancomycin-resistant enterococci (VRE) and avoparcin use became apparent. A recent study regarding faecal samples documented a continuing high prevalence of VRE among Norwegian poultry 3 years after avoparcin was banned. In the present study, carcasses of broilers and turkeys from farms where avoparcin had previously been in use and carcasses of layer chickens from farms where avoparcin had never been used were examined for the presence of VRE. One carcass from each of 150 different farms was included. By a direct plating method, VRE were isolated from 30 of 100 samples of broilers and turkeys, but not from any samples of layer chickens. When an enrichment step was included, VRE were isolated from a total of 81 of the 100 samples of broilers and turkeys and from nine of the 50 samples of layer chickens. All VRE isolated were highly resistant to vancomycin (MIC > or = 256 microg/ml) and possessed the vanA gene. These results correspond to the prevalence of VRE recently documented in faecal samples from Norwegian poultry. The present study reveals a high prevalence of VRE in broiler and turkey carcasses. Consequently, consumers are exposed to VRE when handling raw poultry meat, although the public health significance of such exposure is unclear.
The present descriptive study evaluates breast cancer mortality among women in the northernmost German Federal State of Schleswig-Holstein over the period from 1981 to 1995 based on data provided by the Schleswig-Holstein State Office for Statistics. Starting from the results of the second Cancer Atlas for the Federal Republic of Germany [11], covering the period 1976-1980, breast cancer mortality rates rose sharply until the beginning of our investigation. A moderate rise in breast-cancer mortality in Schleswig-Holstein continued until the third period under investigation (1987-1989), while after that time breast cancer mortality decreased again towards the end of the investigation period. In Schleswig-Holstein as a whole, the mortality rate increased by approximately 12% during the last 20 years. When reviewing age-specific breast-cancer mortality for each rural district ('Kreis') or, respectively, for each self-administrative town (kreisfreie Stadt), different patterns were observed, particularly among younger women. In the 40- to 44-year and 45- to 49-year age group a rise in breast cancer mortality was observed in several, but not all districts. Mortality patterns do not tend to be representative for breast cancer incidence because mortality reflects only a small proportion of the women actually suffering from breast cancer. The standardised incidence of breast cancer in Germany--calculated on the basis of data provided by the Saarland cancer registry--is currently over 60/100,000 and is thus 2-3 times the mortality rate. However, presently it is not possible to evaluate the breast cancer incidence (Schleswig-Holstein) because the first population-based cancer registration was initiated only in 1999. In contrast to other German Federal States, notification of all cancer cases is compulsory in Schleswig-Holstein. A valid analysis of breast cancer incidence should be possible in 5-10 years conditional on a registration rate of at least 90%. Further investigations into the causes of breast cancer should be carried out to test specific hypotheses on exposure to risk factors, including nutrition and life style to clarify the regional differences observed in this study.
We investigated whether early childhood factors such as breast-feeding, parity, and smoking contribute to the variation of organochlorine compounds (OC: dichlorodiphenyldichloroethene, hexachlorobenzene, beta-hexachlorocyclohexane, and the sum of polychlorinated biphenyls including the congeners 101, 118, 138, 153, 170, 180, 183, and 187) at approximately 7 y of age. OC were measured in whole blood of 350 children. Pregnancy characteristics and the child's living conditions were gathered by questionnaires administered to the parents and interviews with the mother. Height and weight were determined during the medical examination. Exclusion of incomplete data and nonbiologic children of the mothers yielded a sample of 337 children. We applied regression analysis with indicator variables, controlling for confounders. No systematic association was detected for birth order or maternal smoking during pregnancy. The OC concentrations are diluted in children with a higher body mass index (>18 kg/m(2)). We found a strong, dose-dependent relationship between the duration of breast-feeding (none, 1-4 wk, 5-8 wk, 9-12 wk, >12 wk) and the concentration of all five OCs. Of the potential determinants analyzed, more of the variance of the OC concentration is accounted for by breast-feeding than by any other variable. Exclusive breast-feeding beyond 12 wk was associated with a doubling of OC whole blood concentration compared with bottle-fed children (dichlorodiphenyldichloroethene, 0.28 microg/L versus 0.55 microg/L; beta-hexachlorocyclohexane, 0.05 microg/L versus 0.14 microg/L; hexachlorobenzene, 0.14 microg/L versus 0.25 microg/L; sum of polychlorinated biphenyls, 0.25 microg/L versus 0.55 microg/L). These findings indicate that breast-feeding can lead to an extended duration of increased OC exposure in childhood.
Five Norwegian broiler farms previously identified as housing broilers carrying vancomycin-resistant enterococci (VRE) were examined for the presence of VRE 4 years after avoparcin was banned. Environmental samples were obtained from empty, cleaned broiler houses. Faecal samples were collected weekly from the flock housed after the environmental sampling. The hatchery from where the chicks originated was also sampled. VRE were found to be present in the farm environment after depopulation and cleanup of the broiler houses. Within 3 weeks after introduction to the farm, all broiler flocks tested positive for VRE. VRE were not isolated from the hatchery.
From October 1997 to April 1998, a survey was conducted to assess the occurrence of pathogenic Yersinia enterocolitica in Norwegian pork products, using a traditional culturing method and a PCR assay. A total of 300 pork samples was examined. Five slaughterhouses in the Norwegian Meat Cooperative were represented with 249 samples and another 51 samples were obtained from retail outlets in the city of Oslo. Using the NMKL method, Y. enterocolitica 0:3 was isolated from six (2%) of the samples, while the PCR method indicated presence of pathogenic Y. enterocolitica in 50 (17%) of the samples. The results indicate that a reduction has occurred in the prevalence of pathogenic Y. enterocolitica in Norwegian pork products, as compared to a previous Norwegian study conducted in 1988-1989. The study also highlights the need for further development and improvement of methods applied for the detection of pathogenic Y. enterocolitica in foods.
Of the gas-phase IR spectrum of nitrylfluoride, FNO(2), between 1200 and 1900 cm(-1), the bands nu(4), 2nu(6), nu(2) + nu(3), and nu(1) have been studied at a resolution of ca. 0.003 cm(-1). Improved ground state rotational and centrifugal distortion constants have been obtained from a simultaneous analysis of the data from F. Hegelund, H. Bürger, and G. Pawelke [J. Mol. Spectrosc. 184, 350-361 (1997)] and the present data from nu(4) and nu(1). The nu(4) and 2nu(6) bands are free from local perturbations, and upper state spectroscopic constants have been obtained from the conventional Watson Hamiltonian. The bands nu(1) and nu(2) + nu(3) are strongly perturbed by Coriolis interactions with the nearby dark levels nu(5) + nu(6), nu(3) + nu(6), and nu(2) + nu(5). Upper state constants for nu(1) and nu(2) + nu(3) are obtained from triad and tetrad models, respectively, including Coriolis resonances within this system. In addition Coriolis interaction parameters and the vibrational energies for the three dark states together with some of their rotational constants are determined from the observed perturbation effects on nu(1) and nu(2) + nu(3). Copyright 2000 Academic Press.
Avoparcin was used as a feed additive in Norwegian broiler and turkey production from 1986 until 1995. It was banned due to the selection of VanA-type vancomycin-resistant enterococci (VRE) in animal husbandry and to reduce the potential for human exposure to VRE. The aim of the present study was to investigate the prevalence of VRE carriage in Norwegian poultry farmers and their poultry three years after avoparcin was banned. Corresponding faecal samples from poultry and humans on farms where avoparcin had previously been used (exposed farms, n = 73) and farms where avoparcin had never been used (unexposed farms, n = 74) were analysed for the presence of VRE. For each farm, one sample from the poultry house and one sample from the farmer were obtained. VRE were isolated from 72 of 73 (99%) and eight of 74 (11%) poultry samples from exposed and unexposed farms, respectively. VRE were isolated from 13 of 73 (18%) and one of 74 (1%) farmer samples from exposed and unexposed farms, respectively. All VRE isolates were highly resistant to vancomycin and possessed the vanA gene, as shown by PCR. The high prevalence of VRE is in accordance with previous Norwegian studies, and shows a remarkable stability of the VanA resistance determinant in an apparently non-selective environment.
This study compares the susceptibility of Enterococcus faecium isolated from pigs and poultry in Denmark, Finland, and Norway to antimicrobial agents used for growth promotion. E. faecium was isolated from 211 broilers and 55 pigs in Denmark in 1997, from Norwegian 55 poultry farms (turkey and broiler farms) and 4 swine farms between 1995 and 1997, and Finnish poultry (52) and swine (43) in 1996 and examined for susceptibility to avilamycin, avoparcin, bacitracin, flavomycin, monensin, salinomycin, spiramycin, tylosin, and virginiamycin. Only a limited number of isolates were categorized as resistant to monensin or salinomycin. In general, an association between the usage of antimicrobial agents in the respective countries and the occurrence of associated resistance was observed. Resistance to avilamycin was frequently observed among isolates from broilers in Denmark, where avilamycin has been used, whereas all isolates from Finland and Norway, where these drugs have not been used, were susceptible. The same phenomenon could be observed for avoparcin, bacitracin, tylosin, and virginiamycin; resistance was frequently observed among isolates from where these antimicrobials have been widely used, but rarely among isolates from where the use has been limited. Also for avoparcin and bacitracin, an association between usage and occurrence of resistance was observed. All isolates categorized as avoparcin resistant contained the vanX gene; isolates from broilers had the T variant in position 8,234 and isolates from pigs the G variant. Three (1%) of the 222 isolates resistant to tylosin contained the ermA gene and 196 (88%) ermB. Sixteen (11%) of the 146 virginiamycin-resistant isolates from broilers and two (7%) of the 27 virginiamycin-resistant isolates from pigs in Denmark contained the satA gene, whereas satA was not observed among any of the virginiamycin-resistant isolates from Finland. A total of 72% of the virginiamycin-resistant E. faecium from broilers in Denmark and all nine virginiamycin-resistant E. faecium from Finland contained satG. This gene was also observed among two (7%) of the virginiamycin-resistant isolates from pigs in Denmark. This study indicates that the use of antimicrobial agents for growth promotion in Denmark, Finland, and Norway have selected for resistance to most of these drugs among E. faecium in food animals.
This study documents a strong and statistically significant association between the use of the glycopeptide avoparcin as a growth promoter in Norwegian poultry production and the occurrence of vancomycin-resistant Enterococcus species (VRE). Avoparcin was approved as a feed additive for broilers and turkeys in Norway in 1986 and was banned from June 1, 1995. In a survey conducted in Norway between June, 1995 and March, 1997, VRE were isolated from fecal samples from 106 out of 109 poultry houses previously exposed to avoparcin (97%) and from six out of 33 poultry houses never exposed to avoparcin (18%) (RR = 5.35). Samples from previously exposed poultry houses were collected in three time periods. The proportion of positive samples remained high (96-98%), in all three time periods indicating a persistence of vancomycin resistance among enterococci for more than a year and a half after the withdrawal of avoparcin. VRE were also isolated from six out of 10 poultry farmers living on farms previously exposed to avoparcin, and from none of 16 farmers living on farms never exposed to avoparcin. Moreover, VRE were isolated from 68 out of the 225 broiler carcasses investigated (30%). The resistance to vancomycin was a high-level type (MIC > or = 256 microg/ml) mediated by the vanA gene. For comparison, VRE could only be isolated from two out of 147 fecal samples from Norwegian flocks of swine (1%). Because avoparcin never has been used in Norwegian swine production, this observation strengthens the association between the use of avoparcin in animal husbandry and the occurrence of VRE.
As part of an epidemiologic study on exposure to a toxic waste incineration plant we investigated whether blood concentrations of polychlorinated biphenyls (PCBs), lead, and cadmium, as well as concentration of mercury in 24-hr urine samples were associated with thyroid hormone status. As an indication of status, we determined levels of thyroid-stimulating hormone (TSH), free thyroxine (FT(4)), and free triiodothyronine (FT(3)) in children living in households where [less than/equal to] 10 cigarettes were smoked per day. Eight PCB congeners (PCBs 101, 118, 138, 153, 170, 180, 183, and 187) were measured in whole blood samples. Of these, seven congeners (PCB 101 was not detected in any sample) and the sum of all PCB congeners were analyzed as predictors for thyroid hormone status in separate linear regression models adjusted for potential confounders. In addition, the possible effects of cadmium, lead, and mercury on levels of thyroid hormones were examined. Blood concentrations and information on questionnaire data were available for 320 children 7-10 years of age. We found a statistically significant positive association between the mono-ortho congener PCB 118 and TSH as well as statistically significant negative relationships of PCBs 138, 153, 180, 183, and 187 to FT(3). There was no association for the PCB congeners and FT(4). Blood cadmium concentration was associated with increasing TSH and diminishing FT(4). Blood lead and urine concentration of mercury were of no importance to thyroid hormone levels. The results stress the need for future studies on the possible influences of PCB and cadmium exposure on thyroid hormones, particularly in children. These studies should also take neurologic development into account.
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