Activity of a dry mist hydrogen peroxide system against environmental Clostridium difficile contamination in elderly care wards

Clostridium difficile causes serious healthcare-associated infections. Infection control is difficult, due in part to environmental contamination with C. difficile spores. These spores are relatively resistant to cleaning and disinfection. The activity of a dry mist hydrogen peroxide decontamination system (Sterinis((R))) against environmental C. difficile contamination was assessed in three elderly care wards. Initial sampling for C. difficile was performed in 16 rooms across a variety of wards and specialties, using Brazier's CCEY (cycloserine-cefoxitin-egg yolk) agar. Ten rooms for elderly patients (eight isolation and two sluice rooms) were then resampled following dry mist hydrogen peroxide decontamination. Representative isolates of C. difficile were typed by polymerase chain reaction ribotyping. C. difficile was recovered from 3%, 11% and 26% of samples from low, medium and high risk rooms, respectively. In 10 high risk elderly care rooms, 24% (48/203) of samples were positive for C. difficile, with a mean of 6.8 colony-forming units (cfu) per 10 samples prior to hydrogen peroxide decontamination. Ribotyping identified the presence of the three main UK epidemic strains (ribotypes 001, 027 and 106) and four rooms contained mixed strains. After a single cycle of hydrogen peroxide decontamination, only 3% (7/203) of samples were positive (P<0.001), with a mean of 0.4 cfu per 10 samples ( approximately 94% reduction). The Sterinis((R)) hydrogen peroxide system significantly reduced the extent of environmental contamination with C. difficile in these elderly care rooms. This relatively quick and user-friendly technology might be a more reliable method of terminally disinfecting isolation rooms, following detergent cleaning, compared to the manual application of other disinfectants.     

Molecular epidemiology of endemic Clostridium difficile infection

This is the first study to provide a comprehensive insight into the molecular epidemiology of endemic Clostridium difficile and particularly that associated with a recently recognized epidemic strain. We DNA fingerprinted all C. difficile isolates from the stools of patients with symptomatic antibiotic-associated diarrhoea and from repeated samples of the inanimate ward environment on two elderly medicine hospital wards over a 22-month period. Notably, C. difficile was not recoverable from either ward immediately before opening, but was found on both wards within 1-3 weeks of opening, and the level of environmental contamination rose markedly during the first 6 months of the study period. C. difficile infection (CDI) incidence data correlated significantly with the prevalence of environmental C. difficile on ward B (r = 0.76, P < 0.05) but not on ward A (r = 0.26, P > 0.05). We found that RAPD and RS-PCR typing had similar discriminatory power, although, despite fingerprinting over 200 C. difficile isolates, we identified only six distinct types. Only two distinct C. difficile strains were identified as causing both patient infection and ward contamination. Attempts to determine whether infected patients or contaminated environments are the prime source for cross-infection by C. difficile had limited success, as over 90% of C. difficile isolates were the UK epidemic clone. However, a non-epidemic strain caused a cluster of six cases of CDI, but was only isolated from the environment after the sixth patient became symptomatic. The initial absence of this strain from the environment implies patient-to-patient and/or staff-to-patient spread. In general, routine cleaning with detergent was unsuccessful at removing C. difficile from the environment. Understanding the epidemiology and virulence of prevalent strains is important if CDI is to be successfully controlled.     

Widespread environmental contamination associated with patients with diarrhea and methicillin-resistant Staphylococcus aureus colonization of the gastrointestinal tract.

OBJECTIVE: Patients colonized with methicillin-resistant Staphylococcus aureus (MRSA) may contaminate their immediate environment with this organism. However, the extent to which gastrointestinal colonization with MRSA affects environmental contamination is not known. We investigated the frequency of environmental contamination in the rooms of patients with diarrheal stools and heavy gastrointestinal colonization with MRSA. DESIGN: Prospective observational study. SETTING: A 500-bed teaching hospital. METHODS: Stool specimens submitted for Clostridium difficile toxin assays were inoculated onto colistin-naladixic acid agar. MRSA was identified with standard methods. Samples from a standardized list of 10 environmental surfaces were cultured, from the rooms of 8 patients who had diarrhea that yielded heavy growth of MRSA (case patients) and from the rooms of 6 MRSA-positive patients with stool cultures negative for MRSA (control patients). MRSA isolates from 13 patients (8 case patients and 5 control patients) and 64 of the environmental isolates recovered from their rooms were compared by pulsed-field gel electrophoresis (PFGE). One clinical isolate from a control patient was excluded because there was no corresponding environmental MRSA isolate with which to compare it. RESULTS: Overall, MRSA were recovered from 47 (58.8%) of 80 surfaces in the rooms of case patients, compared with 14 (23.3%) of 60 surfaces in the rooms of control patients (58.8% [95% CI, 47.8-68.9] vs 23.3% [95% CI, 14.3-35.5]; P<.0001). The items most commonly contaminated were bedside rails, blood pressure cuffs, television remote controls, and toilet seats. Seventy-eight percent of the environmental isolates in patients' rooms had PFGE types that were indistinguishable or closely related to those recovered from the patients' clinical specimens. CONCLUSIONS: Patients who have diarrheal stools and heavy gastrointestinal colonization with MRSA are associated with significantly greater environmental MRSA contamination than patients without MRSA in their stool, and they are likely to be the source of that contamination.     

Tackling contamination of the hospital environment by methicillin-resistant Staphylococcus aureus (MRSA): a comparison between conventional terminal cleaning and hydrogen peroxide vapour decontamination

The hospital environment can sometimes harbour methicillin-resistant Staphylococcus aureus (MRSA) but is not generally regarded as a major source of MRSA infection. We conducted a prospective study in surgical wards of a London teaching hospital affected by MRSA, and compared the effectiveness of standard cleaning with a new method of hydrogen peroxide vapour decontamination. MRSA contamination, measured by surface swabbing was compared before and after terminal cleaning that complied with UK national standards, or hydrogen peroxide vapour decontamination. All isolation rooms, ward bays and bathrooms tested were contaminated with MRSA and several antibiogram types were identified. MRSA was common in sites that might transfer organisms to the hands of staff and was isolated from areas and bed frames used by non-MRSA patients. Seventy-four percent of 359 swabs taken before cleaning yielded MRSA, 70% by direct plating. After cleaning, all areas remained contaminated, with 66% of 124 swabs yielding MRSA, 74% by direct plating. In contrast, after exposing six rooms to hydrogen peroxide vapour, only one of 85 (1.2%) swabs yielded MRSA, by enrichment culture only. The hospital environment can become extensively contaminated with MRSA that is not eliminated by standard cleaning methods. In contrast, hydrogen peroxide vapour decontamination is a highly effective method of eradicating MRSA from rooms, furniture and equipment. Further work is needed to determine the importance of environmental contamination with MRSA and the effect on hospital infection rates of effective decontamination.     

Acquisition of Clostridium difficile from the hospital environment

An outbreak of antibiotic-associated colitis that occurred on a ward of a Michigan hospital during February-April, 1984, was studied by bacteriophage-bacteriocin typing. Stools from the seven involved patients yielded Clostridium difficile isolates of types B1537 or Cld7;B1537. C. difficile was recovered from 31.4% of environmental cultures obtained on the ward, and the majority of isolates were types B1537 or Cld7;B1537. When the ward was disinfected with unbuffered hypochlorite (500 parts per million (ppm) available chlorine), surface contamination decreased to 21% of initial levels and the outbreak subsequently ended. Phosphate buffered hypochlorite (1,600 ppm available chlorine, pH 7.6) was even more effective; its use resulted in a 98% reduction in surface contamination. These findings suggest that environmental contamination with C. difficile is important in the epidemiology of antibiotic-associated colitis, and that hypochlorite is effective in eliminating C. difficile from the hospital environment.     

A molecular characterization of Clostridium difficile isolates from humans, animals and their environments.

It is generally accepted that most patients with Clostridium difficile-associated diarrhoea acquire the organism from the environment. Recently we demonstrated that household pets may constitute a significant reservoir of C. difficile through gastrointestinal carriage in up to 39% of cats and dogs. These findings suggested that direct transmission from household pets, or contamination of the environment by them, may be a factor in the pathogenesis of C. difficile-associated diarrhoea. To investigate this possibility, we examined isolates of C. difficile from humans, pets and the environment by restriction enzyme analysis (REA) and restriction fragment length polymorphism (RFLP) typing using enhanced chemiluminescence. Both REA and RFLP typing methods used Hind III digests of chromosomal DNA. A total of 116 isolates of C. difficile from pets (26), veterinary clinic environmental sites (33), humans (37) and hospital environmental sites (20) was examined. REA was far more discriminatory than RFLP typing and for all isolates there were 34 REA types versus 6 RFLP types. There was good correlation between the REA types found in isolates from pets and from the veterinary clinic environment, and between isolates from humans and from those found in the hospital environment. There was, however, no correlation between REA type of C. difficile found in pets and isolates of human origin. We conclude that there may still be a risk of humans acquiring C. difficile from domestic pets as these findings may be the result of geographical variation.     

Gaseous and air decontamination technologies for Clostridium difficile in the healthcare environment

The recent data for hospital-acquired infections suggest that infection rates for meticillin-resistant Staphylococcus aureus (MRSA) and Clostridium difficile are beginning to decrease. However, while there is still pressure to maintain this trend, the resistance of C. difficile spores to standard detergents continues to present a problem for many UK hospitals trying to prevent its spread or control outbreaks. Alternative disinfection technologies such as gaseous decontamination are currently being marketed to the healthcare sector as an alternative/supplement to manual disinfection, and have been shown to be effective in reducing environmental contamination. When used correctly, they offer a complementary technology to manual cleaning that increases the probability of an effective reduction in viability and provides a comparatively uniform distribution of disinfectant. Three gaseous decontamination technologies are examined for their suitability in reducing environmental contamination with C. difficile: gaseous hydrogen peroxide, chlorine dioxide and ozone. Air decontamination and UV-based technologies are also briefly described. We conclude that while there is a role to play for these new technologies in the decontamination of ward surfaces contaminated with C. difficile, the requirement for both a preclean before use and the limited 'in vivo' evidence means that extensive field trials are necessary to determine their cost-effectiveness in a healthcare setting.     

Value of lysozyme agar incorporation and alkaline thioglycollate exposure for the environmental recovery of Clostridium difficile

Clostridium difficile is an increasingly prevalent nosocomial pathogen. Environmental contamination by spores is believed to be a major factor propagating the spread of C. difficile. Various approaches including the use of bile salts have been described to enhance the recovery of C. difficile from clinical and environmental specimens. We found that lysozyme (5 mg/L) incorporated into a selective medium containing bile salts significantly increased the recovery of C. difficile from swabs of 197 environmental sites (11% versus 24% samples positive, P< 0.01). Furthermore, in a separate series of experiments additional use of cooked meat broth enrichment significantly enhanced the recovery of C. difficile (35% versus 45%, P = 0.009). Conversely, we found that pre-exposure to alkaline thioglycollate did not improve the yield of C. difficile. Lysozyme incorporation markedly increases the recovery of C. difficile from environmental samples probably by stimulation of spore germination. Our findings suggest that previous attempts to determine the level of environmental C. difficile contamination have markedly underestimated the true prevalence of this pathogen.     

Lack of enhanced effect of a chlorine dioxide-based cleaning regimen on environmental contamination with Clostridium difficile spores

Spores of Clostridium difficile may play a significant role in transmission of disease within the healthcare environment and are resistant to a variety of detergents and cleaning fluids. A range of environmental cleaning agents has recently become available, many of which claim to be sporicidal. We investigated the effect of changing to a chlorine dioxide-based cleaning regimen on C.?difficile environmental contamination and patient infection rates. The prevalence of environmental contamination was unaffected with a rate of 8% (9/120) before and 8% (17/212) following the change. Rates of patient infection were also unchanged during these periods.     

Airborne hydrogen peroxide for disinfection of the hospital environment and infection control: a systematic review

We reviewed the effectiveness of airborne hydrogen peroxide as an environmental disinfectant and infection control measure in clinical settings. Systematic review identified ten studies as eligible for inclusion. Hydrogen peroxide was delivered in the form of vapour and dry mist in seven and three studies, respectively. Pathogens evaluated included meticillin-resistant Staphylococcus aureus (MRSA), Clostridium difficile and multiple bacterial types, in five, three, and two studies, respectively. Before the application of any cleaning intervention, 187/480 (39.0%; range: 18.9-81.0%) of all sampled environmental sites were found to be contaminated by the studied pathogens in nine studies that reported specific relevant data. After application of terminal cleaning and airborne hydrogen peroxide, 178/630 (28.3%; range: 11.9-66.1%) of the sampled sites in six studies and 15/682 (2.2%; range: 0-4.0%) of the sampled sites in ten studies, respectively, remained contaminated. Four studies evaluated the use of hydrogen peroxide vapour for infection control. This was associated with control of a nosocomial outbreak in two studies, eradication of persistent environmental contamination with MRSA and decrease in C. difficile infection in each of the remaining two studies.     

Environmental reservoirs of methicillin-resistant Staphylococcus aureus in isolation rooms: correlation with patient isolates and implications for hospital hygiene

Strategies to control and prevent the spread of methicillin-resistant Staphylococcus aureus (MRSA) include early identification of positive patients through screening, patient isolation, hand hygiene, nasal and skin decontamination, and the adequate cleaning and decontamination of clinical areas. However, many national and other guidelines provide few details on environmental decontamination regimens, partly because the role of the environment in the spread of MRSA is not well documented. We prospectively studied the environment of the isolation rooms of 25 MRSA patients for up to four weeks, sampling horizontal surfaces and the air using settle plates as well as an air sampler, while continuing regular daily cleaning according to the hospital protocol. We then typed 20 patient isolates and the corresponding environmental isolates (N=35) to assess the similarity of strains. A high proportion of samples were positive for MRSA; 269/502 (53.6%) surface samples, 70/250 (28%) air samples and 102/251 (40.6%) settle plates. Over half of the surface samples taken from the beds and the mattresses were positive for MRSA. Identical or closely related isolates were recovered from the patient and their environment in 14 (70%) patients, suggesting possible environmental contamination of the isolation rooms, possibly contributing to endemic MRSA. More effective and rigorous use of current approaches to cleaning and decontamination is required as well as consideration of newer technologies to eradicate MRSA and other hospital-acquired pathogens.     

Environmental contamination in child day-care centers

Enteropathogens associated with outbreaks of diarrhea in day-care centers are spread by the fecal-oral route through contaminated hands or environmental objects. This prospective study was undertaken to determine the prevalence of fecal coliform contamination in day care. Ten rooms in six centers housing 121 children less than 2 years of age were studied for 13 weeks (February to May 1988). Inanimate objects (n = 1,275), toy balls (n = 724), and hands (n = 954) were cultured 1-3 times per week. Fecal coliform contamination was common and was greater (p less than 0.05) for objects, toy balls, and hands of children in toddler compared with infant rooms. In five rooms in which clothes were worn over diapers, there was a significantly lower prevalence of fecal coliform contamination of toy balls (p less than 0.005), inanimate objects (p less than 0.05), and hands (p less than 0.001) when compared with rooms in which overclothes were not worn. Occurrence of diarrhea was significantly associated with increased contamination of hands (p = 0.001). Stool and environmental isolates from individual rooms had the same plasmid patterns, which were unique to each center. In summary, fecal coliform contamination of environmental objects and hands of children and caregivers in day-care centers is common; toy balls can serve as sentinels of contamination; fecal coliform contamination can be significantly decreased by use of overclothes; and Escherichia coli strains from stool showed the same plasmid patterns.     

Difference in the incidence of Clostridium difficile among patients infected with human immunodeficiency virus admitted to a public hospital and a private hospital.

OBJECTIVE: To compare the occurrence of Clostridium difficile among inpatients infected with human immunodeficiency virus (HIV) in two different hospitals. DESIGN: Prospective, observational study. SETTING: Specialized HIV inpatient units. PATlENTS: HIV-infected inpatients at Cook County Hospital (CCH) and Rush Presbyterian St. Luke's Medical Center (RPSLMC). INTERVENTIONS: A clinical and epidemiologic assessment of patient risk factors for C. difficile was performed. C. difficile isolates found on stool, rectal, and environmental cultures were typed by pulsed-field gel electrophoresis. RESULTS: Twenty-seven percent of patients admitted to CCH versus 4% of patients admitted to RPSLMC had positive cultures for C. difficile (P = .001). At CCH, 14.7% of environmental cultures were positive versus 2.9% at RPSLMC (P = .002). Risk factors for C. difficile acquisition included hospitalization at CCH, more severe HIV, use of acyclovir and H2-blockers, and longer hospital stay. Patients admitted to CCH were taking more antibiotics, had longer hospital stays, and more frequently had a history of C. difficile infection. During the study, two strains (CD1A and CD4) extensively contaminated the CCH environment. However, only CD1A caused an outbreak. CONCLUSIONS: The C. difficile acquisition rate at CCH was sevenfold higher than that at RPSLMC, and CCH had a more contaminated environment. Differences in patient acquisition rates likely reflect a greater prevalence of traditional C. difficile risk factors and a concurrent outbreak at CCH. Although two strains heavily contaminated the environment at CCH, only one caused an outbreak, suggesting that factors other than the environment are important in initiating C. difficile outbreaks.     

Molecular analysis of relapse vs re-infection in HIV-positive patients suffering from recurrent Clostridium difficile associated diarrhoea.

Recurrence is a major complication of Clostridium difficile associated diarrhoea, especially in human immunodeficiency virus (HIV) positive patients, and it is important to distinguish between relapse and re-infection in recurrent episodes. The aim of our study was to analyse C. difficile isolates obtained from HIV-positive patients with recurrent diarrhoea in order to distinguish between relapse and re-infection. This analysis was based on the study of DNA similarities among isolates obtained from different episodes within each patient. Relapses occurred in 64% of patients, 32% suffered re-infections and a combination of relapse plus re-infection was seen in 4%. DNA typing methods can be useful tools to characterize recurrent episodes of C. difficile associated disease.     

高频接触物体表面清洁质量的调查

目的了解某院高频接触物体表面清洁质量,以制订有效的循证干预措施。方法2014年10月-2015年2月应用荧光胶剂对该院普通病房、手术室、重症监护病房（ICU）高频接触物体表面进行标记,每周1～2次,于患者出院前、手术结束后进行标记,清洁后检查物体表面清洁质量。结果共监测物体表面2 131处,清除1 732处,清除率为81.3%,卫生清洁质量处于合格状态。普通病区监测642处,清除率达82.1%;手术室监测650处,清除率75.8%;ICU区监测839处,清除率84.9%,3个病区物体表面清除率比较,差异有统计学意义（χ2 =19.964,P＜0.001）。普通病区呼叫按钮、电开关和设备带清除率分别是51.3%、55.2%和58.5%,手术室墙和麻醉机控板、摇床器、键盘和输液泵的卫生清除率分别是38.6%、48.8%、60.0%、61.3%、68.4%,ICU设备带的清除率为65.9%,均处于不合格状态。结论该院环境卫生清洁质量总体处于合格状态,但仍有需要改进的地方,该院高频接触物体表面的清洁质量有待进一步提高。     

Risk of bioaerosol contamination with Aspergillus species before and after cleaning in rooms filtered with high-efficiency particulate air filters that house patients with hematologic malignancy.

OBJECTIVE: To examine the impact of cleaning and directional airflow on environmental contamination with Aspergillus species in hospital rooms filtered with high-efficiency particulate air (HEPA) filters that house patients with hematologic malignancy. DESIGN: Detailed environmental assessment. SETTING: A 475-bed tertiary cancer center in the southern United States. METHODS: From April to October 2004, 1,258 surface samples and 627 bioaerosol samples were obtained from 74 HEPA-filtered rooms (in addition, 88 outdoor bioaerosol samples were obtained). Samples were collected from rooms cleaned within 1 hour after patient discharge and from rooms before cleaning. Positive and negative airflows were evaluated using air-current tubes at entrances to patient rooms. RESULTS: Of 1,258 surface samples, 3.3% were positive for Aspergillus species. Univariate analysis showed no relationship between cleaning status and occurrence of Aspergillus species. Of 627 bioaerosol samples, 7.3% were positive for Aspergillus species. Multiple logistic analysis revealed independently significant associations with detection of Aspergillus species. Cleaned rooms positive for Aspergillus species had a higher geometric mean density of colonies than that of rooms sampled before cleaning (18.9 vs 5.5 colony-forming units [cfu] per cubic meter; P=.0047). Rooms with positive airflow had a detection rate for bioaerosol samples equivalent to that of rooms with negative airflow (7.3% vs 7.8%; P=.8). There was no significant difference in the density of Aspergillus species between rooms with negative airflow and rooms with positive airflow (12.5 vs 8.4 cfu/m(3); P=.33). CONCLUSIONS: Concentration of bioaerosol contamination with Aspergillus species was increased in rooms sampled 1 hour after cleaning compared with rooms sampled before cleaning, suggesting a possible correlation between re-entrained bioaerosols (ie, those suspended by activity in the room) after cleaning and the risk of nosocomial invasive aspergillosis.     

Clostridium difficile-associated diarrhoea: epidemiological data from Western Australia.

The incidence of Clostridium difficile-associated diarrhoea (CDAD) was investigated retrospectively at a 690-bed teaching hospital for the period 1983-92. Our aims were to determine: (i) the distribution by age and sex of patients with CDAD, (ii) the possibility of a seasonal trend and, (iii) the influence of infection control procedures, contamination of the hospital environment and the use of third-generation cephalosporins. The laboratory diagnosis of CDAD was based on demonstration of the organism by stool culture and/or detection of specific cytotoxin in stool filtrates. C. difficile was detected in 917 patients who were being investigated for diarrhoeal illness. Yearly isolations varied from a low of 49 in 1983 to a high of 120 in 1990 (Chi square for linear trend 128.8; P < 0.005). Most patients were elderly, with 63% aged 60 years or more; the majority (59%) were female. The relationship between culture of C. difficile and detection of cytotoxin in faecal extracts was also examined. Sixty percent of a sample of 132 isolates from patients in whom faecal cytotoxin was not detected produced cytotoxin in vitro, suggesting that culture is a more sensitive indicator of infection with C. difficile than cytotoxin detection. When the total number of faecal specimens received in the laboratory was used as a denominator there was an increase in the number of incident cases of CDAD between 1983 and 1990, apart from 1986. When occupied bed days was used as the denominator a similar trend was observed with a peak in 1990.(ABSTRACT TRUNCATED AT 250 WORDS)     

Finding a benchmark for monitoring hospital cleanliness

This study evaluated three methods for monitoring hospital cleanliness. The aim was to find a benchmark that could indicate risk to patients from a contaminated environment. We performed visual monitoring, ATP bioluminescence and microbiological screening of five clinical surfaces before and after detergent-based cleaning on two wards over a four-week period. Five additional sites that were not featured in the routine domestic specification were also sampled. Measurements from all three methods were integrated and compared in order to choose appropriate levels for routine monitoring. We found that visual assessment did not reflect ATP values nor environmental contamination with microbial flora including Staphylococcus aureus and meticillin-resistant S. aureus (MRSA). There was a relationship between microbial growth categories and the proportion of ATP values exceeding a chosen benchmark but neither reliably predicted the presence of S. aureus or MRSA. ATP values were occasionally diverse. Detergent-based cleaning reduced levels of organic soil by 32% (95% confidence interval: 16-44%; P<0.001) but did not necessarily eliminate indicator staphylococci, some of which survived the cleaning process. An ATP benchmark value of 100 relative light units offered the closest correlation with microbial growth levels <2.5 cfu/cm(2) (receiver operating characteristic ROC curve sensitivity: 57%; specificity: 57%). In conclusion, microbiological and ATP monitoring confirmed environmental contamination, persistence of hospital pathogens and measured the effect on the environment from current cleaning practices. This study has provided provisional benchmarks to assist with future assessment of hospital cleanliness. Further work is required to refine practical sampling strategy and choice of benchmarks.     

Prevalence of Clostridium difficile in the environment in a rural community in Zimbabwe

Clostridium difficile has been shown to be a nosocomial pathogen associated with diarrhoea and pseudomembranous colitis in hospitalised patients, but very little is known about its prevalence outside the hospital environment. The aim of this study was to determine the prevalence of C. difficile in faeces of domestic animals, soil and drinking water in a rural community. Water, animal faeces and soil were collected from homesteads in a rural community and the samples were cultured for C. difficile.Clostridium difficile isolates that produced toxins A or B were tested for their susceptibility to antimicrobial drugs. Clostridium difficile was isolated from 37.0% of 146 soil samples, 17.4% of 115 chicken faeces samples, 6.0% of 234 water samples and 4.3% of 161 faecal samples of other animals. Some of the C. difficile isolates from chickens (55.0%), soil (66.7%) and water (14.3%) were toxigenic. All toxigenic isolates were susceptible to metronidazole, vancomycin, doxycycline, chloramphenicol and tetracycline and all were resistant to cefotaxime, gentamicin, ciprofloxacin, norfloxacin and nalidixic acid. The results of the present study suggest that chickens kept by villagers are an important reservoir of C. difficile, which may act as a source of human infection.