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P1239 Patients with chronic fatigue syndrome have higher numbers of anaerobic bacteria in the intestine compared to healthy subjects
S340
17th ECCMID / 25th ICC, Posters
University of Pretoria, and a nearby animal shelter with various grades of bite wound. Methods: Fifty dogs with bite wounds inflicted within the previous 72 hours were selected. This represented 104 wounds. Wounds were clinically graded according to severity and evaluated cytologically. Swabs were collected from all wounds for bacterial culture. Wounds were classified as infected or non-infected. Infection was diagnosed if 2 of the following 3 criteria were met: macroscopic purulence, phagocytosed bacteria present or if the wounded dog had pyrexia. Non-infected wounds were either sterile (established by culture) or contaminated (culture positive but bacteria not phagocytosed on cytology). All wounds were cultured aerobically and anaerobically and all aerobic cultures were evaluated for antibiotic susceptibility. Table 1. Percentage antibiotic susceptibility of the most common bacteria
Pasteurellaceae
Staphylococcus intermedius
Pyogenicb streptococci
Escherichia coli
Amoxycillinclavulanate Cloxacillin Penicillin G Cephalothin Ceftiofur Enrofloxacin Orbifloxacin Doxycycline −Sulphamethazole + trimethoprim Gentamicin Amikacin Kanamycin Lincomycin Lincospectin Tylosin
Pasteurella multocida
Susceptibility (%)a
n = 30
n = 13
n = 23
n = 27
n = 10
87
100
91
78
80
64 93 93 93 93 64 93 90
83 92 92 44 85 90 85 100
90 65 100 65 91 74 57 74
70 81 86 81 7 48 67 89
22 10 20 44 50 50 10 60
43 65 83 17 33 76
92 90 92 31 50 92
91 100 95 40 33 95
19 7 11 0 n/a 77
60 89 50 10 0 10
a Shaded
areas indicate susceptibility of 50% or less. streptococci included: S. canis, S. pyogenes, S. agalactiae, Group-C streptococci.
b Pyogenic
Results: Of the 104 wounds, 21 were judged to be infected and 83 non-infected. Seventeen (16%) of all wounds were sterile were also classified as non-infected. This was statistically significant (P = 0.02). Of the 84% that were culture positive, 16% grew aerobes, 1% anaerobes and 67% a mixture of aerobes and anaerobes. A total of 211 isolates were cultured representing a mean of 2.1 isolates per wound. Of the aerobes cultured, 22%, 20% and 17% belonged to Pasteurella, Streptococcus and Staphylococcus species, respectively. Within these groups, Pasteurella multocida (65%) and Staphylococcus intermedius (70%) were predominant. Pasteurella canis and pyogenic streptococci were common in infected wounds, whereas Bacillus spp., Actinomyces spp. and the oral streptococci were usually found in contaminated wounds. Three anaerobic genera were cultured, namely, Prevotella, Clostridium and Peptostreptococcus, and were usually associated with wounds with dead space. This is also the first recorded case of Capnocytophaga canimorsus in an infected dog bite wound. Significantly, clinical examination and
cytological assessment were capable of establishing whether antibiotics were required or not. Although no single antibiotic was considered to be effective against all the bacteria, in vitro, potentiated sulphonamides, ampicillin and amoxycillin plus clavulanic acid gave the best results. P1239 Patients with chronic fatigue syndrome have higher numbers of anaerobic bacteria in the intestine compared to healthy subjects ˚ Sullivan (Stockholm, SE) B. Eveng˚ard, C.E. Nord, A. Objectives: To compare the intestinal microflora in patients with chronic fatigue syndrome with the microflora in healthy subjects. Methods: Ten patients (6 females and 4 men, mean age 38.7, range 30−53 years), fulfilling the criteria according to the US Centers for Disease Control and Prevention (1994) for chronic fatigue syndrome, were included in the study. The patients had high fatigue severity scores and high disability scores. Ten healthy subjects, matched for gender and age, were included as a control group (6 females and 4 men, mean age 37.6, range 29−48 years). None of the patients or the healthy subjects had taken any antimicrobial agents within the preceding 3 months. Two stool samples (2 to 5 days apart) were collected from each subject. The stool specimens were suspended, diluted and inoculated on non-selective and selective media. The aerobic agar plates were incubated for 24 hours and the anaerobic plates for 48 h at 37ºC. After incubation different colony types were counted, isolated in pure cultures and identified to genus level. Streptocoocci and aerobic Gram-negative rods were identified to species level. Median values from the two samples were used in the comparisons. Results: The aerobic genera alpha-haemolytic streptococci, Grampositive bacilli, enterobacteria and Candida as well as anaerobic lactobacilli, bifidobacteria, clostridia, veillonella and bacteroides were found in higher numbers in patients with chronic fatigue syndrome than in healthy subjects. However, only for bifidobacteria the difference was statistically significant (P < 0.05). The difference between the two groups in total numbers of anaerobic microorganisms was also statistically significant (P < 0.001). Conclusion: In this study, higher numbers of anaerobic intestinal microorganisms were identified in patients suffering from chronic fatigue syndrome than in healthy subjects. Disturbances in the intestinal ecology have been implicated as a part of the pathogenesis in chronic fatigue syndrome. Further work is needed to increase our understanding of the role of the intestinal microflora in this disorder. P1240 Detection and characterisation of enterotoxigenic Clostridium perfringens type A isolates in Japanese retail meats Y. Miki, I. Kaneko-Hirano, K. Miyamoto, K. Fujiuchi, S. Akimoto (Wakayama, JP) Objectives: Clostridium perfringens is an important pathogen causing food-borne gastrointestinal disease. In Japan, C. perfringens is ranked as the third or forth greatest cause of the disease and sickened approximately 4,000 people, while the outbreak annually occurred 20 to 40 cases. Previous surveys found that the prevalence of enteropathogenic C. perfringens in Japanese retail food was approximately 0−4%, which was similar to that in USA. It is thought that food poisoning isolates carry cpe on their chromosome, while isolates from other gastrointestinal diseases, such as antibiotic-associated diarrhoea, carry cpe on transferable plasmid. In Japan, recently two food-borne outbreaks by plasmid-cpe isolates were reported, while several C. perfringens food poisoning isolates carry chromosomal cpe. In this survey, we investigated the prevalence of enterotoxigenic C. perfringens in Japanese retail meat samples, and then cpe-positive isolates were tested with multiplex PCR, which could differentiate chromosomal cpe isolate versus plasmid cpe isolate. Methods: We collected two hundred meat samples from grocery stores and meat shops in Wakayama City. Approximately 100 g of food samples were anaerobically incubated at 45ºC overnight with TGC II medium in
17th ECCMID / 25th ICC, Posters
University of Pretoria, and a nearby animal shelter with various grades of bite wound. Methods: Fifty dogs with bite wounds inflicted within the previous 72 hours were selected. This represented 104 wounds. Wounds were clinically graded according to severity and evaluated cytologically. Swabs were collected from all wounds for bacterial culture. Wounds were classified as infected or non-infected. Infection was diagnosed if 2 of the following 3 criteria were met: macroscopic purulence, phagocytosed bacteria present or if the wounded dog had pyrexia. Non-infected wounds were either sterile (established by culture) or contaminated (culture positive but bacteria not phagocytosed on cytology). All wounds were cultured aerobically and anaerobically and all aerobic cultures were evaluated for antibiotic susceptibility. Table 1. Percentage antibiotic susceptibility of the most common bacteria
Pasteurellaceae
Staphylococcus intermedius
Pyogenicb streptococci
Escherichia coli
Amoxycillinclavulanate Cloxacillin Penicillin G Cephalothin Ceftiofur Enrofloxacin Orbifloxacin Doxycycline −Sulphamethazole + trimethoprim Gentamicin Amikacin Kanamycin Lincomycin Lincospectin Tylosin
Pasteurella multocida
Susceptibility (%)a
n = 30
n = 13
n = 23
n = 27
n = 10
87
100
91
78
80
64 93 93 93 93 64 93 90
83 92 92 44 85 90 85 100
90 65 100 65 91 74 57 74
70 81 86 81 7 48 67 89
22 10 20 44 50 50 10 60
43 65 83 17 33 76
92 90 92 31 50 92
91 100 95 40 33 95
19 7 11 0 n/a 77
60 89 50 10 0 10
a Shaded
areas indicate susceptibility of 50% or less. streptococci included: S. canis, S. pyogenes, S. agalactiae, Group-C streptococci.
b Pyogenic
Results: Of the 104 wounds, 21 were judged to be infected and 83 non-infected. Seventeen (16%) of all wounds were sterile were also classified as non-infected. This was statistically significant (P = 0.02). Of the 84% that were culture positive, 16% grew aerobes, 1% anaerobes and 67% a mixture of aerobes and anaerobes. A total of 211 isolates were cultured representing a mean of 2.1 isolates per wound. Of the aerobes cultured, 22%, 20% and 17% belonged to Pasteurella, Streptococcus and Staphylococcus species, respectively. Within these groups, Pasteurella multocida (65%) and Staphylococcus intermedius (70%) were predominant. Pasteurella canis and pyogenic streptococci were common in infected wounds, whereas Bacillus spp., Actinomyces spp. and the oral streptococci were usually found in contaminated wounds. Three anaerobic genera were cultured, namely, Prevotella, Clostridium and Peptostreptococcus, and were usually associated with wounds with dead space. This is also the first recorded case of Capnocytophaga canimorsus in an infected dog bite wound. Significantly, clinical examination and
cytological assessment were capable of establishing whether antibiotics were required or not. Although no single antibiotic was considered to be effective against all the bacteria, in vitro, potentiated sulphonamides, ampicillin and amoxycillin plus clavulanic acid gave the best results. P1239 Patients with chronic fatigue syndrome have higher numbers of anaerobic bacteria in the intestine compared to healthy subjects ˚ Sullivan (Stockholm, SE) B. Eveng˚ard, C.E. Nord, A. Objectives: To compare the intestinal microflora in patients with chronic fatigue syndrome with the microflora in healthy subjects. Methods: Ten patients (6 females and 4 men, mean age 38.7, range 30−53 years), fulfilling the criteria according to the US Centers for Disease Control and Prevention (1994) for chronic fatigue syndrome, were included in the study. The patients had high fatigue severity scores and high disability scores. Ten healthy subjects, matched for gender and age, were included as a control group (6 females and 4 men, mean age 37.6, range 29−48 years). None of the patients or the healthy subjects had taken any antimicrobial agents within the preceding 3 months. Two stool samples (2 to 5 days apart) were collected from each subject. The stool specimens were suspended, diluted and inoculated on non-selective and selective media. The aerobic agar plates were incubated for 24 hours and the anaerobic plates for 48 h at 37ºC. After incubation different colony types were counted, isolated in pure cultures and identified to genus level. Streptocoocci and aerobic Gram-negative rods were identified to species level. Median values from the two samples were used in the comparisons. Results: The aerobic genera alpha-haemolytic streptococci, Grampositive bacilli, enterobacteria and Candida as well as anaerobic lactobacilli, bifidobacteria, clostridia, veillonella and bacteroides were found in higher numbers in patients with chronic fatigue syndrome than in healthy subjects. However, only for bifidobacteria the difference was statistically significant (P < 0.05). The difference between the two groups in total numbers of anaerobic microorganisms was also statistically significant (P < 0.001). Conclusion: In this study, higher numbers of anaerobic intestinal microorganisms were identified in patients suffering from chronic fatigue syndrome than in healthy subjects. Disturbances in the intestinal ecology have been implicated as a part of the pathogenesis in chronic fatigue syndrome. Further work is needed to increase our understanding of the role of the intestinal microflora in this disorder. P1240 Detection and characterisation of enterotoxigenic Clostridium perfringens type A isolates in Japanese retail meats Y. Miki, I. Kaneko-Hirano, K. Miyamoto, K. Fujiuchi, S. Akimoto (Wakayama, JP) Objectives: Clostridium perfringens is an important pathogen causing food-borne gastrointestinal disease. In Japan, C. perfringens is ranked as the third or forth greatest cause of the disease and sickened approximately 4,000 people, while the outbreak annually occurred 20 to 40 cases. Previous surveys found that the prevalence of enteropathogenic C. perfringens in Japanese retail food was approximately 0−4%, which was similar to that in USA. It is thought that food poisoning isolates carry cpe on their chromosome, while isolates from other gastrointestinal diseases, such as antibiotic-associated diarrhoea, carry cpe on transferable plasmid. In Japan, recently two food-borne outbreaks by plasmid-cpe isolates were reported, while several C. perfringens food poisoning isolates carry chromosomal cpe. In this survey, we investigated the prevalence of enterotoxigenic C. perfringens in Japanese retail meat samples, and then cpe-positive isolates were tested with multiplex PCR, which could differentiate chromosomal cpe isolate versus plasmid cpe isolate. Methods: We collected two hundred meat samples from grocery stores and meat shops in Wakayama City. Approximately 100 g of food samples were anaerobically incubated at 45ºC overnight with TGC II medium in