[The microbial threat 2. Monitoring of the connection between the utilization of antibiotics and antibiotic-resistant bacteria in humans and animals].
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Animals↗
Biomedical subjects
H C Wegener
Publications and source records attributed to H C Wegener.
Evaluation and application of ribotyping for epidemiological studies of Actinobacillus pleuropneumoniae in Denmark.
The aim of the present study was to evaluate ribotyping as an epidemiological tool for Actinobacillus pleuropneumoniae and apply the method in studies of A. pleuropneumoniae infections in Danish pig herds. The evaluation of ribotyping was based on the 13 international reference strains and 106 epidemiologically unrelated Danish field strains representing the nine serotypes of biotype 1 (1, 2, 5A/B, 6, 7, 8, 10, 12, and K2:O7) and one serotype 14 of biotype 2. Enzymes CfoI and HindIII were chosen for generation of ribotype patterns. Ribotyping of the reference strains resulted in 10 CfoI types and 11 HindIII types. Ribotyping of the Danish strains resulted in 17 different CfoI ribotypes and 24 different HindIII ribotypes. Combining HindIII- and CfoI-ribotyping divided the Danish strains into 26 different types. The stability, reproducibility and typability of ribotype patterns were good, and the discriminatory power was between 0.85-0.89. The relatively low discriminatory power was caused by four predominant types, containing 61% of the isolates. The typing system was applied in studies of routes of infection of specific pathogen-free (SPF) pig herds and included 112 strains of A. pleuropneumoniae. Airborne transmission from neighboring conventional pig farms was investigated in 12 cases of infected SPF herds. Transmission via vehicles transporting pigs between SPF herds was investigated in nine cases while transmission by trading of pigs between SPF herds was investigated in two cases. Serotype 2 was isolated from all SPF herds included in this study, except one, emphasizing the high prevalence of this serotype in Denmark. By ribotyping, airborne transmission was indicated in five of 12 cases, transmission via pig transporting vehicle was indicated in six of nine cases, and transmission via trading was indicated in one of two cases. In many cases findings of predominant ribotypes made interpretations of suspected routes of transmission difficult. The relationship of strains based on ribotypes was calculated using Dices coefficient and clustered by UPGMA. HindIII ribotypes of serotype 2 strains were closely related, though only showing 43% similarity to HindIII ribotypes of remaining serotypes.
Actinobacillus Infections↗
Resistance to antimicrobial agents used for animal therapy in pathogenic-, zoonotic- and indicator bacteria isolated from different food animals in Denmark: a baseline study for the Danish Integrated Antimicrobial Resistance Monitoring Programme (DANMAP).
This study describes the establishment and first results of a continuous surveillance system of antimicrobial resistance among bacteria isolated from pigs, cattle and broilers in Denmark. The three categories of bacteria tested were: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter coli/jejuni, Salmonella enterica, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). A total of 3304 bacterial isolates collected from October 1995 through December 1996 were tested for susceptibility to all major classes of antimicrobial agents used for therapy in Denmark. Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Acquired resistance to all antimicrobials was found. The occurrence of resistance varied by animal origin and bacterial species. In general, resistance was observed more frequently among isolates from pigs than from cattle and broilers. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed, as is the occurrence of resistance in other countries. The results of this study show the present level of resistance to antimicrobial agents among a number of bacterial species isolated from food animals in Denmark. Thus, the baseline for comparison with future prospective studies has been established, enabling the determination of trends over time.
Animal Diseases↗
Surveillance of antimicrobial resistance in bacteria isolated from food animals to antimicrobial growth promoters and related therapeutic agents in Denmark.
This study was conducted to describe the occurrence of acquired resistance to antimicrobials used for growth promotion among bacteria isolated from swine, cattle and poultry in Denmark. Resistance to structurally related therapeutic agents was also examined. Three categories of bacteria were tested: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter, Salmonella, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). All antimicrobials used as growth promoters in Denmark and some structurally related therapeutic agents (in brackets) were included: Avilamycin, avoparcin (vancomycin), bacitracin, carbadox, flavomycin, monensin, olaquindox, salinomycin, spiramycin (erythromycin, lincomycin), tylosin (erythromycin, lincomycin), and virginiamycin (pristinamycin). Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Breakpoints for growth promoters were established by population distribution of the bacteria tested. A total of 2,372 bacterial isolates collected during October 1995 to September 1996 were included in the study. Acquired resistance to all currently used growth promoting antimicrobials was found. A frequent occurrence of resistance were observed to avilamycin, avoparcin, bacitracin, flavomycin, spiramycin, tylosin and virginiamycin, whereas resistance to carbadox, monensin, olaquindox and salinomycin was less frequent. The occurrence of resistance varied by animal origin and bacterial species. The highest levels of resistance was observed among enterococci, whereas less resistance was observed among zoonotic bacteria and bacteria pathogenic to animals. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed. The results show the present level of resistance to growth promoters in bacteria from food animals in Denmark. They will form the baseline for comparison with future prospective studies, thereby enabling the determination of trends over time.
Animals↗
Historical yearly usage of glycopeptides for animals and humans: the American-European paradox revisited.
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Animals↗
Isolation of vancomycin resistant Enterococcus faecium from food.
In a survey of vancomycin resistant Enterococcus faecium (VREF) in Danish meat products, VREF could be detected in 16% of 160 samples of broilers collected at slaughterhouses and in 15% of 26 samples of pork collected from the retail trade. VREF were isolated by enrichment for 24 h in nutrient broth supplemented with vancomycin (50 micrograms/ml) prior to plating on Slanetz and Bartley agar. Using direct plating on Slanetz and Bartley agar, VREF could be isolated from only 1.7% of 540 samples of broilers from slaughterhouses and 2.2% of 90 samples of broilers from retail outlets. VREF was not detected in 124 samples of pork and 128 samples of beef from retail outlets by the direct plating method. An additional enrichment step in nutrient broth supplemented with vancomycin enhanced the detection rate of VREF by approximately three times compared to the direct plating method when investigating the same 160 samples of broilers by the two methods. The implications and public health aspects of VREF in food is discussed.
Animals↗
Staphylococcus hyicus exfoliative toxin: purification and demonstration of antigenic diversity among toxins from virulent strains.
The exfoliative toxin produced by Staphylococcus hyicus strain 1289D-88 was purified as a single protein of approximately 30 kDa. Extracellular proteins of S. hyicus grown under small scale fermentation conditions were precipitated with ammonium sulfate. Separation of proteins was performed by hydrophobic interaction chromatography and successively anion exchange chromatography. The purified toxin was tested in a piglet skin assay. Weak epidermal lesions were macroscopically and microscopically similar to lesions caused by (NH4)2SO4-precipitated culture supernatant from the same strain. Addition of 0.5 mM CuSo4 to the purified toxin resulted in more intense skin alterations comparable to lesions caused by precipitated culture supernatant diluted 1:10. These results indicated that the activity of the exfoliative toxin was dependent on the presence of Cu2+. Polyclonal and monoclonal antibodies were prepared against the exfoliative toxin from strain 1289D-88. The in vivo activity of the exfoliative toxin could be neutralized by antibodies. It was shown that polyclonal as well as monoclonal antibodies only reacted with the toxin produced by two of nine well-defined virulent strains of S. hyicus. These results showed antigenic diversity among exfoliative toxins produced by different strains of S. hyicus.
Animals↗
Antimicrobial resistance in Salmonella enterica subsp. enterica serovar typhimurium from humans and production animals.
We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent to the State Serum Institute during August 1993 (228 isolates). The animal strains were isolated from clinical or subclinical infections in cattle (48 isolates), pigs (99 isolates) or poultry (98 isolates), all from 1993. All strains were tested against 22 different antimicrobial agents used in both human and veterinary medicine with the tablet diffusion method. Strains were also phage-typed and the plasmid content determined in all resistant strains. Ribotyping was performed on selected strains. Of 228 human isolates tested, 19.3% of the strains were resistant to one or more antimicrobial agent compared with 10.4% of strains from cattle, 11.1% of strains from pigs and 9.2% of strains from poultry. Multiple resistance, i.e. resistance against at least four antimicrobial agents, was found in 9.2% of the human strains, but in only two of the cattle isolates. The majority of the multi-resistant strains in humans were from infections contracted outside Denmark, most often in southern Europe or south-east Asia. Resistance in human strains was most common against tetracycline (13%), ampicillin (12%), sulphonamide (12%), streptomycin (10%) and chloramphenicol (8%). The resistance pattern differed somewhat in animal isolates: Poultry strains were usually resistant only to ampicillin, while pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance was present in S. typhimurium isolated from humans and animals in Denmark, multiple resistance was most often acquired outside Denmark.
Animals↗
Correlation of conversion of Salmonella enterica serovar enteritidis phage type 1, 4, or 6 to phage type 7 with loss of lipopolysaccharide.
Studies of pairs of Salmonella enterica serovar Enteritidis isolates from different poultry flocks showed that phage type (PT) 7 may be derived from PT 1, 4, and 6. The conversion appeared to be associated with loss of lipopolysaccharide. It is concluded that PT 7 may be of little value as an epidemiological marker of S. enterica serovar Enteritidis.
Humans↗
Evaluation of a fluorescence-labelled oligonucleotide probe targeting 23S rRNA for in situ detection of Salmonella serovars in paraffin-embedded tissue sections and their rapid identification in bacterial smears.
A method for the detection of Salmonella based on fluorescence in situ hybridization (FISH) has been developed and applied for the direct detection of Salmonella in pure cultures and in formalin-fixed, paraffin-embedded tissue sections. On the basis of the 23S rRNA gene sequences representing all of the S. enterica subspecies and S. bongori, an 18-mer oligonucleotide probe was selected. The specificity of the probe was tested by in situ hybridization to bacterial cell smears of pure cultures. Forty-nine of 55 tested Salmonella serovars belonging to subspecies I, II, IIIb, IV, and VI hybridized with the probe. The probe did not hybridize to serovars from subspecies IIIa (S. arizonae) or to S. bongori. No cross-reaction to 64 other strains of the family Enterobacteriaceae or 18 other bacterial strains outside this family was observed. The probe was tested with sections of formalin-fixed, paraffin-embedded tissue from experimentally infected mice or from animals with a history of clinical salmonellosis. In these tissue sections the probe hybridized specifically to Salmonella serovars, allowing for the detection of single bacterial cells. The development of a fluorescence-labelled specific oligonucleotide probe makes the FISH technique a promising tool for the rapid identification of S. enterica in bacterial smears, as well as for the detection of S. enterica in histological tissue sections.
Animals↗
Comparison of conventional culture methods and two commercial enzyme immunoassays for detection of Salmonella in porcine fecal samples and cecal contents.
Two commercial enzyme immunoassays, designated EIA-1 and EIA-2, for the detection of salmonella in feces and cecal contents were compared to conventional culture methods. Out of 362 cecal content samples, 35 were positive by EIA-1 and 30 were positive by EIA-2 and conventional methods. Out of 189 fecal samples, 41 were positive by EIA-1, 30 were positive by EIA-2, and 24 were positive by conventional culture methods. The EIA-1 assay method detected more positive samples overall, although only comparison of the fecal samples was statistically significant. The results indicate that the EIA-1 method is rapid, requiring only 24 hours, and may be more sensitive than the EIA-2 assay and conventional culture techniques. Enzyme immunoassays for the detection of salmonella in fecal samples or cecal contents provide a reasonable alternative to conventional culture techniques.
Animals↗
A study of phage- and ribotype patterns of Staphylococcus aureus isolated from bovine mastitis in the Nordic countries.
This study was conducted to investigate the geographical distribution of phage and ribotypes of Staphylococcus aureus causing bovine mastitis in the 5 Nordic countries. A total of 403 isolates of S. aureus was isolated from 403 different dairy herds. One hundred five strains were isolated in Denmark, 81 in Finland, 17 in Iceland, 96 in Norway and 104 in Sweden. The isolates were phage typed and characterized for their EcoRI restriction fragment length polymorphisms of the genes encoding ribosomal RNA (ribotyping). A total of 351 (87%) of the 403 isolates could be typed by phages assigning them to 25 different phage types. Two to 3 different phage types predominated within each country. One type (29/52) accounted for 36% of all the isolates and was found in 4 of the countries. A total of 87 different ribotypes was found among the isolates investigated. As for phage typing 2 to 3 different types predominated within countries. However, except for one type (ribotype 1), which was commonly found in Denmark, Sweden and Finland, different ribotypes predominated within each country. The combination of phage and ribotyping assigned the isolates to 178 different types. Ninety-six percent of the isolates of ribotype 1 belonged to phage type 29/52. This combined type accounted for 17% of all the 403 isolates. These findings show that a large number of different types of S. aureus can be isolated from cases of bovine mastitis. However, few types predominate within different countries. These predominating types seem to be specific in each country, however, a single type was common for both Denmark, Sweden and Finland. This could suggest differences in the virulence or in modes of transmission of predominating and rare types of S. aureus associated with bovine mastitis.
Animals↗
Association between production of fibrinolysin and virulence of Staphylococcus hyicus in relation to exudative epidermitis in pigs.
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Animals↗
Public health and pork and pork products: regional perspectives of Denmark.
An ambitious programme to eliminate pork as an important source of human salmonellosis was initiated in Denmark in 1993 by the Ministry of Food, Agriculture and Fisheries. The programme comprises control of feedmills, breeding and multiplying herds, slaughter herds and slaughter plants, as well as the final product, fresh pork. As a consequence, the level of occurrence of Salmonella spp. in fresh pork produced in Denmark is approximately 1%. Yersinia enterocolitica 0:3 infections are common in slaughter pig herds in Denmark, and pork is considered to be the only source of human infection in the country. The incidence of pork-related occurrences of human salmonellosis and yersiniosis in 1996 was approximately nine cases per 100,000 inhabitants for both diseases. All swine in Denmark are screened for Trichinella spp. infection, although no positive results have been obtained since 1930. Swine are not considered to be a source for Campylobacter jejuni or enterohaemorrhagic Escherichia coli in Denmark. Listeria monocytogenes can be detected in relatively high rates in pork: however, the incidence of human listeriosis is only 0.5 cases per 100,000 inhabitants. Toxoplasma gondii antibodies have been demonstrated in 3% of slaughter pigs, though the importance of pork as a source of infection is probably very low. Denmark is officially free from Brucella abortus, B. melitensis and Mycobacterium bovis.
Abattoirs↗
Listeria monocytogenes in bovine mastitis. Possible implication for human health.
During the 23-year period 1972 through 1994 quarter milk samples from 1,132,958 cows originating from 36199 herds were examined for the presence of Listeria monocytogenes. Through the period the reference population amounted to 12,742,600 cow years and 401,682 herd years. The percentage of cows infected with L. monocytogenes varied from 0.01 to 0.1% (mean 0.04%) and of herds with an infected cow from 0.2 to 4.2% (mean 1.2%) through the period, showing a low but constant level of infection. A comparison of 33 isolates from bovine mastitis and 27 human clinical isolates was made by sero- and ribotyping. Serotyping showed that all bovine and 17 (63%) of the human isolates belonged to serogroup 1, whereas 10 (37%) of the human isolates belonged to serogroup 4. Ribotyping using EcoRI as restriction enzyme divided the 60 isolates into 16 different types, 7 of which were found among both the bovine and human types. The combination of the typing methods showed that 26 (79%) bovine and 13 (48%) human isolates shared common types. This study showed that a low but constant percentage of Danish dairy herds have cows infected with L. monocytogenes and that some of the bovine types could be found among types causing human infections.
Animals↗
Investigation of an outbreak of human salmonellosis caused by Salmonella enterica ssp. enterica serovar Infantis by use of pulsed field gel electrophoresis.
Analysis of chromosomal DNA restriction patterns produced by pulsed field gel electrophoresis (PFGE) was used to investigate an outbreak of human salmonellosis caused by Salmonella enterica ssp. enterica serovar Infantis (S. Infantis) involving more than 500 registered human cases. The outbreak had been tentatively traced back to a single pig slaughterhouse. A total of 135 isolates from various sources produced 21 different PFGE patterns with the restriction endonuclease XbaI. All human isolates from the outbreak belonged to a single type, the 'EPI-type', whereas human isolates recovered before and after the outbreak belonged to several different types. All isolates investigated from the suspect pig slaughterhouse and its supplier pig herds belonged to the EPI-type. Isolates from pork from the central meat market in Copenhagen, which received most of the carcasses from the suspect slaughterhouse, also belonged to the EPI-type. This was furthermore, the case for isolates from beef from the same market, indicating that cross-contamination had taken place. All isolates from pork and some, but not all, isolates from beef, collected in butchers' shops during the outbreak belonged to the EPI-type. The typing results supported that the outbreak was a common source outbreak, probably originating from a limited number of supplier pig herds supplying animals to a single slaughterhouse.
Abattoirs↗
Listeria monocytogenes in poultry and poultry products: epidemiological investigations in seven Danish abattoirs.
Listeria monocytogenes was isolated from 11/236 (4 x 7%) caecal samples from parent flocks, providing broilers to the abattoirs investigated. Caecal samples from 2078 broilers representing 90 randomly selected broiler flocks were negative for L. monocytogenes. A total of 3080 samples from seven abattoirs including poultry processing line samples, and final products were also examined for L. monocytogenes. Listeria monocytogenes was isolated in 0 x 3% to 18 x 7% of the samples collected in the different abattoirs. Epidemiological typing of 247 L. monocytogenes isolates, including serotyping, phage typing, pulsed-field gel electrophoresis and ribotyping revealed 62 different clones. Based upon typability and discriminatory power, DNA typing methods used were found equally suitable as epidemiological markers. Serotyping and phage typing were not found useful as epidemiological markers for poultry isolates of L. monocytogenes since only 120/247 (48 x 6%) isolates were typable by phage typing and 230/247 (93 x 1%) L. monocytogenes belonged to serotype 01 while 6/247 (2 x 4%) belonged to 04. The discovery of a few dominating clones in each abattoir might indicate an endemic occurrence of L. monocytogenes. It is concluded that L. monocytogenes in the broiler production is primarily localized to the abattoirs. The incidence of L. monocytogenes may be reduced by improving the hygiene.
Abattoirs↗
Typing of Salmonella enterica serovar Saintpaul: an outbreak investigation.
During the summer of 1993 an outbreak of human salmonellosis caused by Salmonella serovar Saintpaul occurred in Denmark. A total of 35 isolates originating from pigs, turkeys and imported foodstuffs, and 10 human isolates were compared following their characterization by agglutination of the 0:5 factor, antibiogram typing, plasmid profiling, ribotyping and pulsed field gel electrophoresis, in order to identify the most probable source of infection. After typing, the source of the investigated outbreak remains obscure because so far no isolates with traits of the outbreak strain have been recovered from production animals. Presence of the 0:5 factor and absence of plasmids in human and porcine isolates pointed to pork as the source of infection, whereas human isolates and all Danish isolates from turkeys had the same ribotype, indicating that turkey was the infection source. A possible explanation for the failure to find isolates with traits of the outbreak strain could be the presence of a third, but so far unidentified, source. The present investigation illustrates the necessity of using more than one epidemiological typing method for outbreak investigation. This is especially important when the organism involved is relatively uncommon and little is known about its diversity and distribution.
Animals↗