- Email: [email protected]
P1240 Detection and characterisation of enterotoxigenic Clostridium perfringens type A isolates in Japanese retail meats
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
Community-acquired bacterial infections stomacher bag, and then cultured samples were tested for the presence of cpe-positive C. perfringens followed by further incubation on SFP agar plate for bacterial isolation. DNA were prepared from overnight culture with InstaGene matrix (BioRad) and then used for PCR reaction for cpe detection. Results: In PCR reaction, we detected cpe from 8 samples (4%), from three of which, we isolated cpe-positive C. perfringens. Interestingly, all three cpe-positive isolates carry cpe on the plasmid and it is combined with downstream IS1470-like structure in PCR cpe-genotyping assay. Conclusion: From these results, C. perfringens carrying cpe on the plasmid might be a possible pathogen of C. perfringens food poisoning in Japan. P1241 Isolation and characterisation of Salmonella from Turkish avian food samples D. Avsaroglu, M. Jaber, M. Akcelik, F. Bozoglu, A. Schroeter, B. Guerra, R. Helmuth (Ankara, TR; Berlin, DE) Objective: To assess the prevalence of Salmonella (S.) in avian food samples from Turkey and to characterise the strains according to their serotypes, phage types and antimicrobial resistance (R) patterns. Methods: 76 chicken meat samples were collected from different markets in Ankara, Turkey (2005–2006). Strains were isolated according to ISO 6579. Biochemical tests were performed with the API20E system. Serological analyses, and phage typing of S. Enteritidis and S. Typhimurium, were carried out. All isolates were tested for their susceptibility to 16 antimicrobial agents by the disk diffusion method. All resistant isolates were further tested by broth microdilution and for an extra panel of 11 b-lactams, as well. Results: 45% of the chicken meat samples analysed (34 out of 76) were positive for Salmonella. From these samples, 71 isolates were obtained. After phenotypic characterisation, they were considered as 42 epidemiologically unrelated strains. The 42 strains belonged to 12 different serotypes. The most prevalent was S. Enteritidis (10 strains, 24%) followed by S. Infantis and S. Virchow (7, 17% each); S. Group C1 with antigenic formula [6,7: k: -] (5); S. Kentucky, S. ssp. I rough form (3); S. Thompson (2); S. Agona, S. Corvallis, S. Nchanga, S. Senftenberg, and S. Typhimurium (1). Among S. Enteritidis, phage type PT21 was predominant (6 of 10), followed by PT6, PT1 and PT3 (2, 1, and 1 strain). Forty per cent (17 strains) of the strains were resistant to some antimicrobial agent. From these, 13 strains were multiresistant and four monoresistant. The most frequent resistance was to nalidixic acid (36% of all strains, 88% of the resistant). All these strains showed also decreased susceptibility to ciprofloxacin. Multidrug resistance to more than 6 antimicrobials was observed in S. Infantis (6 strains showed KAN-NEO-NAL-STR-SPESUL-TET-TMP-SXT resistance), and S. Virchow (1 strain AMP-CEFNAL-STR-SUL-TMP-SXT). This Virchow strain showed resistance to several extended-spectrum b-lactams. All S. Enteritidis were susceptible, except one PT21 NAL-resistant strain. Conclusion: Almost half of the food samples analysed were contaminated with Salmonella, especially serotypes S. Enteritidis, S. Infantis and S. Virchow. Antibiotic resistance is also a problem in the contaminated food from avian origin (40% resistance). The quinolone resistance in all serotypes and multi-resistance in S. Infantis and S. Virchow need further attention. P1242 Is a cholera outbreak a natural disaster? Overview on a cholera outbreak in Iran in 2005 N. Jonaidi jafari, M. Radfar, H. Ghofrani, M. Izadi (Tehran, IR) Objectives: There has been a sharp increase in the number of cholera cases reported to WHO during 2005. A total of 131,943 cases, including 2,272 deaths, have been notified from 52 countries. Overall, this represents a 30% increase compared with the number of cases reported in 2004. During the summer of 2005, an outbreak of choler a struck Iran, infecting 1,118 individuals and killing 11 patients. The epidemic
S341 started from the Southern regions and rapidly disseminated across the country. Methods: In this descriptive, epidemiological research, the related data about all individuals Diagnosed with cholera during the epidemic of summer 2005 has been gathered from the Disease Management Center (DMC). The number of patients in each province, their demographic data, and the death toll were included. A stool sample from all patients with the clinical diagnosis of cholera was sent to the provincial medical laboratory for confirmation, serotyping, and antibiogram. All confirmed samples were then sent to the central reference laboratory in Tehran for further confirmation. If both these tests were positive for V. cholera, the patient was considered a confirmed case. Results: It is difficult to pinpoint the first case of the recent outbreak. In 25 June 2005, a 26 month-old Iranian child was admitted in the Southern city of Bushehr with a possible diagnosis of cholera. Ten days later, two Pakistanis were reported to have the same symptoms in the Southeastern city of Chabahar, near the Iran-Pakistan border. Serological studies showed that the former was caused by O1 biotype El Tor serotype Ogawa, while in the two latter cases, the Inab a serotype was isolated. Most cases were reported in Tehran (216 cases), Hamadan (187 cases) and Qom (152 cases) provinces – all located in central Iran. Almost all Iranian provinces were struck by the epidemic. The total number of confirmed cases at the end of the epidemic was 1,118, of which 11 had died. Conclusion: There is clear trend that cholera is re-emerging in parallel with the ever-increasing proportion of vulnerable populations who live in unsanitary situations. Globally cholera remains a threat for social development. Almost all of the developing countries in the out world are facing either a cholera outbreak or the threat of an epidemic. P1243 Contamination of toothbrushes with Helicobacter pylori, a possible source of infection transmission A. Helaly (Alexandria, EG) Introduction: Helicobacter pylori (HP) infection is a common aetiology in some gastric disorders such as peptic ulcers. The route of infection transmission is not clear. The natural reservoir for HP is not known. HP in dental plaque and saliva had been detected with nested PCR. Objectives: The aim of this work was to examine the presence of the organism in the toothbrushes of patients with positive infection and compare that with controls without HP infection, in an attempt to search for a possible source of reservoir. Methods: Forty volunteer patients (10 female, 30 male, mean age 41.2 years) with a positive Helicobacter pylori (HP) infection were included in the study, and ten negative HP volunteers of matched age and sex. All patients had positive test for HP during routine endoscopic examination, those with positive test and those with negative test (biopsy from the antrum) were included in the study. The toothbrush (at least used for one weeks by the patients for 3 times per day) was rinsed in 50 mL of 0.9% NaCl for about 30 minutes, then centrifugation was done, the sediment was examined for HP by CLO test. The result was analysed using standard statistical analysis and student t table. Result: positive test is considered when any sign of the presence of the infection is present. Positive test was in 28 patients with positive HP infection, no one was positive in the negative volunteers (P > 0.005). Conclusion: toothbrushes may be a source of HP infection, so that the use of a new one is essential during HP treatment to avoid re-infection. Also the change of toothbrushes is a good hygienic habit as regards HP infection. There may be a possible role for anti HP infection solution to handle the toothbrushes with it.
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
Community-acquired bacterial infections stomacher bag, and then cultured samples were tested for the presence of cpe-positive C. perfringens followed by further incubation on SFP agar plate for bacterial isolation. DNA were prepared from overnight culture with InstaGene matrix (BioRad) and then used for PCR reaction for cpe detection. Results: In PCR reaction, we detected cpe from 8 samples (4%), from three of which, we isolated cpe-positive C. perfringens. Interestingly, all three cpe-positive isolates carry cpe on the plasmid and it is combined with downstream IS1470-like structure in PCR cpe-genotyping assay. Conclusion: From these results, C. perfringens carrying cpe on the plasmid might be a possible pathogen of C. perfringens food poisoning in Japan. P1241 Isolation and characterisation of Salmonella from Turkish avian food samples D. Avsaroglu, M. Jaber, M. Akcelik, F. Bozoglu, A. Schroeter, B. Guerra, R. Helmuth (Ankara, TR; Berlin, DE) Objective: To assess the prevalence of Salmonella (S.) in avian food samples from Turkey and to characterise the strains according to their serotypes, phage types and antimicrobial resistance (R) patterns. Methods: 76 chicken meat samples were collected from different markets in Ankara, Turkey (2005–2006). Strains were isolated according to ISO 6579. Biochemical tests were performed with the API20E system. Serological analyses, and phage typing of S. Enteritidis and S. Typhimurium, were carried out. All isolates were tested for their susceptibility to 16 antimicrobial agents by the disk diffusion method. All resistant isolates were further tested by broth microdilution and for an extra panel of 11 b-lactams, as well. Results: 45% of the chicken meat samples analysed (34 out of 76) were positive for Salmonella. From these samples, 71 isolates were obtained. After phenotypic characterisation, they were considered as 42 epidemiologically unrelated strains. The 42 strains belonged to 12 different serotypes. The most prevalent was S. Enteritidis (10 strains, 24%) followed by S. Infantis and S. Virchow (7, 17% each); S. Group C1 with antigenic formula [6,7: k: -] (5); S. Kentucky, S. ssp. I rough form (3); S. Thompson (2); S. Agona, S. Corvallis, S. Nchanga, S. Senftenberg, and S. Typhimurium (1). Among S. Enteritidis, phage type PT21 was predominant (6 of 10), followed by PT6, PT1 and PT3 (2, 1, and 1 strain). Forty per cent (17 strains) of the strains were resistant to some antimicrobial agent. From these, 13 strains were multiresistant and four monoresistant. The most frequent resistance was to nalidixic acid (36% of all strains, 88% of the resistant). All these strains showed also decreased susceptibility to ciprofloxacin. Multidrug resistance to more than 6 antimicrobials was observed in S. Infantis (6 strains showed KAN-NEO-NAL-STR-SPESUL-TET-TMP-SXT resistance), and S. Virchow (1 strain AMP-CEFNAL-STR-SUL-TMP-SXT). This Virchow strain showed resistance to several extended-spectrum b-lactams. All S. Enteritidis were susceptible, except one PT21 NAL-resistant strain. Conclusion: Almost half of the food samples analysed were contaminated with Salmonella, especially serotypes S. Enteritidis, S. Infantis and S. Virchow. Antibiotic resistance is also a problem in the contaminated food from avian origin (40% resistance). The quinolone resistance in all serotypes and multi-resistance in S. Infantis and S. Virchow need further attention. P1242 Is a cholera outbreak a natural disaster? Overview on a cholera outbreak in Iran in 2005 N. Jonaidi jafari, M. Radfar, H. Ghofrani, M. Izadi (Tehran, IR) Objectives: There has been a sharp increase in the number of cholera cases reported to WHO during 2005. A total of 131,943 cases, including 2,272 deaths, have been notified from 52 countries. Overall, this represents a 30% increase compared with the number of cases reported in 2004. During the summer of 2005, an outbreak of choler a struck Iran, infecting 1,118 individuals and killing 11 patients. The epidemic
S341 started from the Southern regions and rapidly disseminated across the country. Methods: In this descriptive, epidemiological research, the related data about all individuals Diagnosed with cholera during the epidemic of summer 2005 has been gathered from the Disease Management Center (DMC). The number of patients in each province, their demographic data, and the death toll were included. A stool sample from all patients with the clinical diagnosis of cholera was sent to the provincial medical laboratory for confirmation, serotyping, and antibiogram. All confirmed samples were then sent to the central reference laboratory in Tehran for further confirmation. If both these tests were positive for V. cholera, the patient was considered a confirmed case. Results: It is difficult to pinpoint the first case of the recent outbreak. In 25 June 2005, a 26 month-old Iranian child was admitted in the Southern city of Bushehr with a possible diagnosis of cholera. Ten days later, two Pakistanis were reported to have the same symptoms in the Southeastern city of Chabahar, near the Iran-Pakistan border. Serological studies showed that the former was caused by O1 biotype El Tor serotype Ogawa, while in the two latter cases, the Inab a serotype was isolated. Most cases were reported in Tehran (216 cases), Hamadan (187 cases) and Qom (152 cases) provinces – all located in central Iran. Almost all Iranian provinces were struck by the epidemic. The total number of confirmed cases at the end of the epidemic was 1,118, of which 11 had died. Conclusion: There is clear trend that cholera is re-emerging in parallel with the ever-increasing proportion of vulnerable populations who live in unsanitary situations. Globally cholera remains a threat for social development. Almost all of the developing countries in the out world are facing either a cholera outbreak or the threat of an epidemic. P1243 Contamination of toothbrushes with Helicobacter pylori, a possible source of infection transmission A. Helaly (Alexandria, EG) Introduction: Helicobacter pylori (HP) infection is a common aetiology in some gastric disorders such as peptic ulcers. The route of infection transmission is not clear. The natural reservoir for HP is not known. HP in dental plaque and saliva had been detected with nested PCR. Objectives: The aim of this work was to examine the presence of the organism in the toothbrushes of patients with positive infection and compare that with controls without HP infection, in an attempt to search for a possible source of reservoir. Methods: Forty volunteer patients (10 female, 30 male, mean age 41.2 years) with a positive Helicobacter pylori (HP) infection were included in the study, and ten negative HP volunteers of matched age and sex. All patients had positive test for HP during routine endoscopic examination, those with positive test and those with negative test (biopsy from the antrum) were included in the study. The toothbrush (at least used for one weeks by the patients for 3 times per day) was rinsed in 50 mL of 0.9% NaCl for about 30 minutes, then centrifugation was done, the sediment was examined for HP by CLO test. The result was analysed using standard statistical analysis and student t table. Result: positive test is considered when any sign of the presence of the infection is present. Positive test was in 28 patients with positive HP infection, no one was positive in the negative volunteers (P > 0.005). Conclusion: toothbrushes may be a source of HP infection, so that the use of a new one is essential during HP treatment to avoid re-infection. Also the change of toothbrushes is a good hygienic habit as regards HP infection. There may be a possible role for anti HP infection solution to handle the toothbrushes with it.