Som resultat af projekterne kan drages folgende hovedkonklusioner om kompostering af faeces fra m... more Som resultat af projekterne kan drages folgende hovedkonklusioner om kompostering af faeces fra mennesker: Faeces fra de undersogte typer af kompostbeholdere bor ikke anvendes til jordbrugsformal uden viderebehandling, da dette skonnes at vaere behaeftet med hygiejne- og sundhedsrisici. Det skyldes, at der ikke blev dokumenteret egentlige termofile temperaturstigninger i faecesmaterialet i komposttoiletenhederne i Hjortshoj, Dyssekilde og Sverige, samt at faekale indikatorbakteriers antal varierede voldsomt, og der ingen entydig tendens var til forekomst af lavere kimtal ved lange opbevaringstider af de opsamlede faekalier.
We present molecular evidence that a distinct genotype of Vibrio cholerae O1 which appeared in Ca... more We present molecular evidence that a distinct genotype of Vibrio cholerae O1 which appeared in Calcutta, India, in September 1993 and which is characterized by a unique ribotype that is not found in the standardized ribotyping scheme of V. cholerae and that shows a specific pulsed-field gel electrophoresis profile may have spread to the west African country of Guinea-Bissau where it was responsible for an epidemic of cholera which began in October 1994 and continued into 1996.
Representative encapsulated strains of Vibrio vulnificus from market oysters and oyster-associate... more Representative encapsulated strains of Vibrio vulnificus from market oysters and oyster-associated primary septicemia cases (25 isolates each) were tested in a blinded fashion for potential virulence markers that may distinguish strains from these two sources. These isolates were analyzed for plasmid content, for the presence of a 460-bp amplicon by randomly amplified polymorphic DNA PCR, and for virulence in subcutaneously (s.c.) inoculated, iron-dextran-treated mice. Similar percentages of market oyster and clinical isolates possessed detectable plasmids (24 and 36%, respectively), produced the 460-bp amplicon (45 and 50%, respectively), and were judged to be virulent in the mouse s.c. inoculation-iron-dextran model (88% for each). Therefore, it appears that nearly all V. vulnificus strains in oysters are virulent and that genetic tests for plasmids and specific PCR size amplicons cannot distinguish between fully virulent and less virulent strains or between clinical and environme...
The risk for contamination of potatoes and groundwater through subsurface drip irrigation with lo... more The risk for contamination of potatoes and groundwater through subsurface drip irrigation with low quality water was explored in 30 large-scale lysimeters containing repacked coarse sand and sandy loam soils. The human pathogens, Salmonella Senftenberg, Campylobacter jejuni and Escherichia coli O157:H7, and the virus indicator Salmonella Typhimurium bacteriophage 28B, were added weekly through irrigation tubes for one month with low irrigation rates (8 mm per week). In the following six months lysimeters were irrigated with groundwater free of pathogens. Two weeks after irrigation was started, phage 28B was detected in low concentrations (2 pfu ml(-1)) in leachate from both sandy loam soil and coarse sand lysimeters. After 27 days, phage 28B continued to be present in similar concentrations in leachate from lysimeters containing coarse sand, while no phage were found in lysimeters with sandy loam soil. The added bacterial pathogens were not found in any leachate samples during the entire study period of 212 days. Under the study conditions with repacked soil, limited macropores and low water velocity, bacterial pathogens seemed to be retained in the soil matrix and died-off before leaching to groundwater. However, viruses may leach to groundwater and represent a health risk as for some viruses only few virus particles are needed to cause human disease. The bacterial pathogens and the phage 28B were found on the potato samples harvested just after the application of microbial tracers was terminated. The findings of bacterial pathogens and phage 28 on all potato samples suggest that the main risk associated with subsurface drip irrigation with low quality water is faecal contamination of root crops, in particular those consumed raw.
... Af Anita Forslund, Carsten Suhr Jacobsen, Ole Stig Jacobsen og Anders Dalsgaard ... af smitst... more ... Af Anita Forslund, Carsten Suhr Jacobsen, Ole Stig Jacobsen og Anders Dalsgaard ... af smitstoffer gennem jordsøjler duktionen gennem optimering af markvanding, og ved at bruge alternative vandkilder, herunder lavkvalitetsvand, som kan være overfladevand fra åer, søer mv. ...
Sixteen clinical Vibrio cholerae O1 strains from four different countries were selected for compa... more Sixteen clinical Vibrio cholerae O1 strains from four different countries were selected for comparison by traditional ribotyping and an automated RiboPrinter system for identification and discrimination purposes. Automated ribotyping, which routinely uses the restriction enzyme EcoRI for typing all bacterial species, produced only five different ribotypes compared with 10 different EcoRI ribotypes obtained by the traditional method. Traditional and automated ribotyping using the restriction enzyme BglI, which is recommended for the ribotyping of V. cholerae, produced 10 and seven different ribotypes, respectively. The lower discrimination shown by the RiboPrinter system was caused mainly by an inability to differentiate closely located fragments due to a lower resolution and electrophoresis conditions, a parameter which cannot be changed in the automated system. The RiboPrinter system includes a database for bacterial identification. However, none of the V. cholerae O1 strains studied showed EcoRI ribotype patterns which matched any of the patterns included in the database. In conclusion, the existing RiboPrinter system is not adequate for taxonomic identification and classification of V. cholerae O1.
The amount of animal manure used in modern agriculture is increasing due to the increase in globa... more The amount of animal manure used in modern agriculture is increasing due to the increase in global animal production. Pig slurry is known to contain zoonotic bacteria such as E. coli, Salmonella spp. and Campylobacter spp., and viruses such as hepatitis E virus and group A rotavirus. Coliform bacteria, present in manure, have previously been shown to leach into tile drains. This poses a potential threat to aquatic environments and may also influence the quality of drinking water. As knowledge is especially scarce about the fate of viruses when applied to fields in natural settings, this project sets out to investigate the leaching potential of six different microorganisms: E. coli and Enterococcus spp. (detected by colony assay), somatic coliphages (using plaque assays), and hepatitis E virus, porcine circovirus type 2, and group A rotavirus (by real-time polymerase chain reaction). All six microorganisms leached through the soil entering the tile drains situated at 1-m depth the first day following pig slurry application. The leaching pattern of group A rotavirus differed substantially from the pattern for somatic coliphages, which are otherwise used as indicators for virus contamination. Furthermore, group A rotavirus was detected in monitoring wells at 3.5-m depth up to 2 months after pig slurry application. The detection of viral genomic material in drainage water and shallow groundwater signifies a potential hazard to human health that needs to be investigated further, as water reservoirs used for recreational use and drinking water are potentially contaminated with zoonotic pathogens.
Abstract The safe use of treated domestic wastewater for irrigation needs to address the risks fo... more Abstract The safe use of treated domestic wastewater for irrigation needs to address the risks for humans (workers, exposed via contact with irrigation water, soil, crops and food, consumers, exposed via ingestion of fresh and processed food), for animals (via ingestion of crops an soil), for the crops and agricultural productivity (via salinity and trace element uptake), for soil (via accumulation or release of pollutants) as well as for surface, groundwaters and the associated ecosystems (via runoff and infiltration, Kass et al., 2005, ...
The presence and genetic content of integrons were investigated for 37 epidemiologically unrelate... more The presence and genetic content of integrons were investigated for 37 epidemiologically unrelated multiple-drug-resistant strains of Salmonella enterica serotype Typhimurium from humans. All isolates were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, sulfonamides, and trimethoprim, as well as to tetracycline and/or nalidixic acid; 20% of them were also resistant to gentamicin and amikacin. Three different class 1 integrons (In-t1, In-t2, and In-t3) were identified by Southern blot hybridization, PCR, and DNA sequencing, and these integrons were found to carry the aadB, catB3, oxa1, aadA1a, aacA4, and aacC1 gene cassettes. Integrons In-t1 (aadB and catB3) and In-t2 (oxa1 and aadA1a) were both located on a conjugative IncFI plasmid of 140 kb. In-t3 (aacA4, aacC1, and aadAIa) was located on an IncL/M plasmid of 100 kb which was present, in association with the IncFI plasmid, in gentamicin- and amikacin-resistant isolates. Despite the extensive similarity at the le...
Seventy clinical strains of Vibrio cholerae O1 isolated from 1982-96 in Samutsakorn, a port city ... more Seventy clinical strains of Vibrio cholerae O1 isolated from 1982-96 in Samutsakorn, a port city 30 km southwest of Bangkok where cholera occurs at low levels with regular seasonality, were characterized to investigate if there were any differences among the O1 strains isolated before, during and after the 0139 epidemic. Pulsed-field gel electrophoresis (PFGE) typing, ribotyping and southern blot hybridization with a cholera toxin probe (CT genotyping) demonstrated several genotypes among O1 strains isolated before the emergence of V. cholerae 0139. However, O1 strains isolated during and after the advent of 0139 showed identical ribotypes which were distinctly different from the types identified in strains isolated before the emergence of 0139. Ribotypes identified in strains during and after the advent of 0139 were also demonstrated by O1 strains isolated immediately before the emergence of 0139. Considering the seasonality of cholera in Samutsakorn, the identical ribotype and CT ...
The number of cholera cases and the mortality rates reported from different regions of Vietnam va... more The number of cholera cases and the mortality rates reported from different regions of Vietnam varied considerably in the period from 1979 to 1996, with between 2,500 and 6,000 cases reported annually from 1992 to 1995. Annual mortality rates ranged from 2.0 to 9.6% from 1979 to 1983 to less than 1.8% after 1983. Major cholera outbreaks were reported from the High Plateau region for the first time in 1994 and 1995; this is an area with limited access to health services and safe drinking-water supplies. All cases were associated with Vibrio cholerae O1. Using ribotyping, cholera toxin (CT) genotyping, and characterization of antibiotic susceptibility patterns and antibiotic resistance genes by PCR, we show that strains isolated after 1990 were clearly different from strains isolated before 1991. In contrast to strains isolated before 1991, 94% of 104 strains isolated after 1990 showed an identical ribotype R1, were resistant to sulfamethoxazole and streptomycin, and showed a differen...
Som resultat af projekterne kan drages folgende hovedkonklusioner om kompostering af faeces fra m... more Som resultat af projekterne kan drages folgende hovedkonklusioner om kompostering af faeces fra mennesker: Faeces fra de undersogte typer af kompostbeholdere bor ikke anvendes til jordbrugsformal uden viderebehandling, da dette skonnes at vaere behaeftet med hygiejne- og sundhedsrisici. Det skyldes, at der ikke blev dokumenteret egentlige termofile temperaturstigninger i faecesmaterialet i komposttoiletenhederne i Hjortshoj, Dyssekilde og Sverige, samt at faekale indikatorbakteriers antal varierede voldsomt, og der ingen entydig tendens var til forekomst af lavere kimtal ved lange opbevaringstider af de opsamlede faekalier.
We present molecular evidence that a distinct genotype of Vibrio cholerae O1 which appeared in Ca... more We present molecular evidence that a distinct genotype of Vibrio cholerae O1 which appeared in Calcutta, India, in September 1993 and which is characterized by a unique ribotype that is not found in the standardized ribotyping scheme of V. cholerae and that shows a specific pulsed-field gel electrophoresis profile may have spread to the west African country of Guinea-Bissau where it was responsible for an epidemic of cholera which began in October 1994 and continued into 1996.
Representative encapsulated strains of Vibrio vulnificus from market oysters and oyster-associate... more Representative encapsulated strains of Vibrio vulnificus from market oysters and oyster-associated primary septicemia cases (25 isolates each) were tested in a blinded fashion for potential virulence markers that may distinguish strains from these two sources. These isolates were analyzed for plasmid content, for the presence of a 460-bp amplicon by randomly amplified polymorphic DNA PCR, and for virulence in subcutaneously (s.c.) inoculated, iron-dextran-treated mice. Similar percentages of market oyster and clinical isolates possessed detectable plasmids (24 and 36%, respectively), produced the 460-bp amplicon (45 and 50%, respectively), and were judged to be virulent in the mouse s.c. inoculation-iron-dextran model (88% for each). Therefore, it appears that nearly all V. vulnificus strains in oysters are virulent and that genetic tests for plasmids and specific PCR size amplicons cannot distinguish between fully virulent and less virulent strains or between clinical and environme...
The risk for contamination of potatoes and groundwater through subsurface drip irrigation with lo... more The risk for contamination of potatoes and groundwater through subsurface drip irrigation with low quality water was explored in 30 large-scale lysimeters containing repacked coarse sand and sandy loam soils. The human pathogens, Salmonella Senftenberg, Campylobacter jejuni and Escherichia coli O157:H7, and the virus indicator Salmonella Typhimurium bacteriophage 28B, were added weekly through irrigation tubes for one month with low irrigation rates (8 mm per week). In the following six months lysimeters were irrigated with groundwater free of pathogens. Two weeks after irrigation was started, phage 28B was detected in low concentrations (2 pfu ml(-1)) in leachate from both sandy loam soil and coarse sand lysimeters. After 27 days, phage 28B continued to be present in similar concentrations in leachate from lysimeters containing coarse sand, while no phage were found in lysimeters with sandy loam soil. The added bacterial pathogens were not found in any leachate samples during the entire study period of 212 days. Under the study conditions with repacked soil, limited macropores and low water velocity, bacterial pathogens seemed to be retained in the soil matrix and died-off before leaching to groundwater. However, viruses may leach to groundwater and represent a health risk as for some viruses only few virus particles are needed to cause human disease. The bacterial pathogens and the phage 28B were found on the potato samples harvested just after the application of microbial tracers was terminated. The findings of bacterial pathogens and phage 28 on all potato samples suggest that the main risk associated with subsurface drip irrigation with low quality water is faecal contamination of root crops, in particular those consumed raw.
... Af Anita Forslund, Carsten Suhr Jacobsen, Ole Stig Jacobsen og Anders Dalsgaard ... af smitst... more ... Af Anita Forslund, Carsten Suhr Jacobsen, Ole Stig Jacobsen og Anders Dalsgaard ... af smitstoffer gennem jordsøjler duktionen gennem optimering af markvanding, og ved at bruge alternative vandkilder, herunder lavkvalitetsvand, som kan være overfladevand fra åer, søer mv. ...
Sixteen clinical Vibrio cholerae O1 strains from four different countries were selected for compa... more Sixteen clinical Vibrio cholerae O1 strains from four different countries were selected for comparison by traditional ribotyping and an automated RiboPrinter system for identification and discrimination purposes. Automated ribotyping, which routinely uses the restriction enzyme EcoRI for typing all bacterial species, produced only five different ribotypes compared with 10 different EcoRI ribotypes obtained by the traditional method. Traditional and automated ribotyping using the restriction enzyme BglI, which is recommended for the ribotyping of V. cholerae, produced 10 and seven different ribotypes, respectively. The lower discrimination shown by the RiboPrinter system was caused mainly by an inability to differentiate closely located fragments due to a lower resolution and electrophoresis conditions, a parameter which cannot be changed in the automated system. The RiboPrinter system includes a database for bacterial identification. However, none of the V. cholerae O1 strains studied showed EcoRI ribotype patterns which matched any of the patterns included in the database. In conclusion, the existing RiboPrinter system is not adequate for taxonomic identification and classification of V. cholerae O1.
The amount of animal manure used in modern agriculture is increasing due to the increase in globa... more The amount of animal manure used in modern agriculture is increasing due to the increase in global animal production. Pig slurry is known to contain zoonotic bacteria such as E. coli, Salmonella spp. and Campylobacter spp., and viruses such as hepatitis E virus and group A rotavirus. Coliform bacteria, present in manure, have previously been shown to leach into tile drains. This poses a potential threat to aquatic environments and may also influence the quality of drinking water. As knowledge is especially scarce about the fate of viruses when applied to fields in natural settings, this project sets out to investigate the leaching potential of six different microorganisms: E. coli and Enterococcus spp. (detected by colony assay), somatic coliphages (using plaque assays), and hepatitis E virus, porcine circovirus type 2, and group A rotavirus (by real-time polymerase chain reaction). All six microorganisms leached through the soil entering the tile drains situated at 1-m depth the first day following pig slurry application. The leaching pattern of group A rotavirus differed substantially from the pattern for somatic coliphages, which are otherwise used as indicators for virus contamination. Furthermore, group A rotavirus was detected in monitoring wells at 3.5-m depth up to 2 months after pig slurry application. The detection of viral genomic material in drainage water and shallow groundwater signifies a potential hazard to human health that needs to be investigated further, as water reservoirs used for recreational use and drinking water are potentially contaminated with zoonotic pathogens.
Abstract The safe use of treated domestic wastewater for irrigation needs to address the risks fo... more Abstract The safe use of treated domestic wastewater for irrigation needs to address the risks for humans (workers, exposed via contact with irrigation water, soil, crops and food, consumers, exposed via ingestion of fresh and processed food), for animals (via ingestion of crops an soil), for the crops and agricultural productivity (via salinity and trace element uptake), for soil (via accumulation or release of pollutants) as well as for surface, groundwaters and the associated ecosystems (via runoff and infiltration, Kass et al., 2005, ...
The presence and genetic content of integrons were investigated for 37 epidemiologically unrelate... more The presence and genetic content of integrons were investigated for 37 epidemiologically unrelated multiple-drug-resistant strains of Salmonella enterica serotype Typhimurium from humans. All isolates were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, sulfonamides, and trimethoprim, as well as to tetracycline and/or nalidixic acid; 20% of them were also resistant to gentamicin and amikacin. Three different class 1 integrons (In-t1, In-t2, and In-t3) were identified by Southern blot hybridization, PCR, and DNA sequencing, and these integrons were found to carry the aadB, catB3, oxa1, aadA1a, aacA4, and aacC1 gene cassettes. Integrons In-t1 (aadB and catB3) and In-t2 (oxa1 and aadA1a) were both located on a conjugative IncFI plasmid of 140 kb. In-t3 (aacA4, aacC1, and aadAIa) was located on an IncL/M plasmid of 100 kb which was present, in association with the IncFI plasmid, in gentamicin- and amikacin-resistant isolates. Despite the extensive similarity at the le...
Seventy clinical strains of Vibrio cholerae O1 isolated from 1982-96 in Samutsakorn, a port city ... more Seventy clinical strains of Vibrio cholerae O1 isolated from 1982-96 in Samutsakorn, a port city 30 km southwest of Bangkok where cholera occurs at low levels with regular seasonality, were characterized to investigate if there were any differences among the O1 strains isolated before, during and after the 0139 epidemic. Pulsed-field gel electrophoresis (PFGE) typing, ribotyping and southern blot hybridization with a cholera toxin probe (CT genotyping) demonstrated several genotypes among O1 strains isolated before the emergence of V. cholerae 0139. However, O1 strains isolated during and after the advent of 0139 showed identical ribotypes which were distinctly different from the types identified in strains isolated before the emergence of 0139. Ribotypes identified in strains during and after the advent of 0139 were also demonstrated by O1 strains isolated immediately before the emergence of 0139. Considering the seasonality of cholera in Samutsakorn, the identical ribotype and CT ...
The number of cholera cases and the mortality rates reported from different regions of Vietnam va... more The number of cholera cases and the mortality rates reported from different regions of Vietnam varied considerably in the period from 1979 to 1996, with between 2,500 and 6,000 cases reported annually from 1992 to 1995. Annual mortality rates ranged from 2.0 to 9.6% from 1979 to 1983 to less than 1.8% after 1983. Major cholera outbreaks were reported from the High Plateau region for the first time in 1994 and 1995; this is an area with limited access to health services and safe drinking-water supplies. All cases were associated with Vibrio cholerae O1. Using ribotyping, cholera toxin (CT) genotyping, and characterization of antibiotic susceptibility patterns and antibiotic resistance genes by PCR, we show that strains isolated after 1990 were clearly different from strains isolated before 1991. In contrast to strains isolated before 1991, 94% of 104 strains isolated after 1990 showed an identical ribotype R1, were resistant to sulfamethoxazole and streptomycin, and showed a differen...
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