Danilo Lo Fo Wong

Global Monitoring of Salmonella Serovar Distribution from the World Health Organization Global Foodborne Infections Network Country Data Bank: Results of Quality Assured Laboratories from 2001 to 2007

Salmonella enterica is commonly acquired from contaminated food and is an important cause of illness worldwide. Interventions are needed to control Salmonella; subtyping Salmonella by serotyping is useful for targeting such interventions. We, therefore, analyzed the global distribution of the 15 most frequently identified serovars of Salmonella isolated from humans from 2001 to 2007 in laboratories from 37 countries that participated in World Health Organization Global Foodborne Infections Network and demonstrated serotyping proficiency in the Global Foodborne Infections Network External Quality Assurance System. In all regions throughout the study period, with the exception of the Oceania and North American regions, Salmonella serovars Enteritidis and Typhimurium ranked as the most common and second most common serovar, respectively. In the North American and Oceania (Australia and New Zealand) regions, Salmonella serovar Typhimurium was the most common serovar reported, and Salmonella serovar Enteritidis was the second most common serovar. During the study period, the proportion of Salmonella isolates reported from humans that were Salmonella serovar Enteritidis was 43.5% (range: 40.6% [2007] to 44.9% [2003]), and Salmonella serovar Typhimurium was 17.1% (range: 15% [2007] to 18.9% [2001]). Salmonella serovars Newport (mainly observed in Latin and North American and European countries), Infantis (dominating in all regions), Virchow (mainly observed in Asian, European, and Oceanic countries), Hadar (profound in European countries), and Agona (intense in Latin and North American and European countries) were also frequently isolated with an overall proportion of 3.5%, 1.8%, 1.5%, 1.5%, and 0.8%, respectively. There were large differences in the most commonly isolated serovars between regions, but lesser differences between countries within the same region. The results also highlight the complexity of the global epidemiology of Salmonella and the need and importance for improving monitoring data of those serovars of highest epidemiologic importance.

Web-based Surveillance and Global Salmonella Distribution, 2000–2002

Surveillance improves control of Salmonella infections.

Salmonella serovars from humans and other sources in Thailand, 1993-2002.

We serotyped 44,087 Salmonella isolates from humans and 26,148 from other sources from 1993 through 2002. The most common serovar causing human salmonellosis in Thailand was Salmonella enterica Weltevreden. Serovars causing human infections in Thailand differ from those in other countries and seem to be related to Salmonella serovars in different food products and reservoirs.

Integrating genome-based informatics to modernize global disease monitoring, information sharing, and response.

The rapid advancement of genome technologies holds great promise for improving the quality and speed of clinical and public health laboratory investigations and for decreasing their cost. The latest generation of genome DNA sequencers can provide highly detailed and robust information on disease-causing microbes, and in the near future these technologies will be suitable for routine use in national, regional, and global public health laboratories. With additional improvements in instrumentation, these next- or third-generation sequencers are likely to replace conventional culture-based and molecular typing methods to provide point-of-care clinical diagnosis and other essential information for quicker and better treatment of patients. Provided there is free-sharing of information by all clinical and public health laboratories, these genomic tools could spawn a global system of linked databases of pathogen genomes that would ensure more efficient detection, prevention, and control of endemic, emerging, and other infectious disease outbreaks worldwide.

Laboratory-Based Surveillance of Nontyphoidal Salmonella Infections in China

Foodborne infections are an important public health problem in China. In 2008, we conducted surveillance for laboratory-confirmed nontyphoidal Salmonella to monitor trends for this infection in China and to build capacity for rapid detection and response to foodborne outbreaks. Salmonella isolates from patients with diarrhea were sent from hospitals to local public health laboratories for confirmation, serotyping, and antimicrobial susceptibility testing. A total of 126 hospitals in 44 cities and counties from 8 provinces provided isolates and epidemiologic data for analysis. Of 23,140 stool specimens submitted to clinical laboratories, 662 (3%) grew Salmonella enterica. Salmonella were most commonly detected between April and October. The median age of infected patients was 27 years; 34% of infections occurred in patients <5 years old. Of the 662 isolates, we found 73 serotypes, of which serotype Enteritidis (31%) and serotype Typhimurium (26%) were the most common. The prevalence of resistance was high for clinically important antimicrobial agents, including ampicillin (41%) and ciprofloxacin (6%). More than 60% of isolates, including 35% of all Typhimurium, were resistant to three or more antimicrobial agents. In this first multiprovince surveillance report of laboratory-confirmed Salmonella infections in China, we found that Enteritidis and Typhimurium are the most common serotypes and that efforts to reduce antimicrobial resistance among Salmonella in China are needed. Although no outbreaks were detected using this system, efforts to improve this systems capacity to do so are underway.

Reproducible Infection Model for Clostridium perfringens in Broiler Chickens

Abstract Experiments were carried out to establish an infection and disease model for Clostridium perfringens in broiler chickens. Previous experiments had failed to induce disease and only a transient colonization with challenge strains had been obtained. In the present study, two series of experiments were conducted, each involving four groups of chickens with each group kept in separate isolators. A coccidial vaccine given at 10 times the prescribed dosage was used to promote the development of necrotic enteritis. In the first experiment, cultures of C. perfringens were mixed with the feed at day 9, 10, 11, and 12, and the coccidial vaccine was given at day 10, whereas in the second experiment, C. perfringens cultures were mixed with the feed at day 17, 18, 19, and 20, and the coccidial vaccine was given at day 18. Chickens were examined at day 9, 11, 12, and 15 (Experiment 1), and at day 17, 18, 20, and 24 (Experiment 2). There was no mortality in any of the groups; however, chickens in the groups receiving both coccidial vaccine and C. perfringens developed the subclinical form of necrotic enteritis, demonstrated by focal necroses in the small intestine, whereas chickens in control groups or groups receiving only coccidial vaccine or only C. perfringens cultures developed no necroses. The results underline the importance of predisposing factors in the development of necrotic enteritis.

Shigella from Humans in Thailand During 1993 to 2006: Spatial-Time Trends in Species and Serotype Distribution

In Thailand during 1993-2006, a total of 9063 Shigella isolates from different medical centers were serotyped and trends over time and spatial clustering analyzed. Of 3583 cases with age information, 1315 (37%) cases were from children between 0 and 4 years and 684 (19%) from children between 5 and 8 years. Most infections were recorded during 1993-1994 (> 1500 per year), decreasing to < 200 in 2006. The relative species distribution also changed. During 1993-1994, Shigella flexneri accounted for 2241 (65%) of 3474 isolations. This proportion decreased to 64 (36%) of 176 infections in 2006. Most infections occurred during July and August, and fewest in December. S. flexneri clustered around Bangkok, and Shigella sonnei in southern Thailand. Most S. flexneri infections were caused by serotype 2a (1590 of 4035) followed by serotype var X (1249). For both serotypes, a pronounced decrease in the number of isolates occurred over time. A much smaller decrease was observed for serotype 3a isolates. Phase I S. sonnei was initially most common, but shifted gradually over phase I, II, to only phase II. No differences in spatial distribution were found. The three most common S. flexneri serotypes all clustered in, around, and west of Bangkok. Serotypes 2a and 3a also clustered in southern Thailand, whereas var X clustered north and northeast of Bangkok. In conclusion, looking at Shigella species, Thailand changed from being a developing country to a developed country between 1995 and 1996. In addition, major shifts in the types of S. sonnei were observed as were differences in spatial clustering of S. flexneri and S. sonnei and S. flexneri serotypes.

Results of Use of WHO Global Salm-Surv External Quality Assurance System for Antimicrobial Susceptibility Testing of Salmonella Isolates from 2000 to 2007

ABSTRACT An international External Quality Assurance System (EQAS) for antimicrobial susceptibility testing of Salmonella was initiated in 2000 by the World Health Organization (WHO) Global Salm-Surv in order to enhance the capacities of national reference laboratories to obtain reliable data for surveillance purposes worldwide. Seven EQAS iterations have been conducted from 2000 to 2007. In each iteration, participating laboratories submitted susceptibility results from 10 to 15 antimicrobial agents for eight Salmonella isolates and an Escherichia coli reference strain (ATCC 25922). A total of 287 laboratories in 102 countries participated in at least one EQAS iteration. A large number of laboratories reported results for the E. coli ATCC 25922 reference strain which were outside the quality control ranges. Critical deviations for susceptibility testing of the Salmonella isolates varied from 4% in 2000 to 3% in 2007. Consistent difficulties were observed in susceptibility testing of amoxicillin-clavulanic acid, cefotaxime, ceftazidime, streptomycin, sulfonamides, and tetracycline. Regional variations in performance were observed, with laboratories in central Asia, Africa, and the Middle East not performing as well as those in other regions. Results from the WHO Global Salm-Surv EQAS show that most laboratories worldwide are capable of correctly performing antimicrobial susceptibility testing of Salmonella isolates, but they also indicate that further improvement for some laboratories is needed. In particular, further training and dissemination of information on quality control, appropriate interpretive criteria (breakpoints), and harmonization of the methodology worldwide through WHO Global Salm-Surv and other programs will contribute to the generation of comparable and reliable antimicrobial susceptibility data (D. M. A. Lo Fo Wong, R. S. Hendriksen, D. J. Mevius, K. T. Veldman, and F. M. Aarestrup, Vet. Microbiol. 115:128-139, 2006).

Association between tetracycline consumption and tetracycline resistance in Escherichia coli from healthy Danish slaughter pigs.

It has been recognized that exposure to antimicrobial agents can exert a selective pressure for the emergence of antimicrobial resistance. The objective of this study was to investigate an association between the probability of isolating a tetracycline-resistant Escherichia coli isolate from the intestinal tract of healthy pigs and patterns of tetracycline consumption in the herds of origin, together with other risk factors. Data on antimicrobial resistance, antimicrobial consumption, and pig herd demographics were obtained from different Danish surveillance programs. Descriptive statistics were performed for the risk factors in relation to the susceptibility status. Logistic regression analysis was performed to identify risk factors with significant effect on the log odds of tetracycline resistance of E. coli isolates. The model showed that an increase in the interval between last prescription and sampling date would decrease the probability of isolating a resistant E. coli isolate (p-value = 0.01). Also, a direct association between treatment incidence rate in a herd and probability of resistance was detected (p-value = 0.03). Other risk factors found to have a significant effect in the isolate susceptibility status were number of produced animals in the year and year of sampling. Other antimicrobial consumption risk factors, such as number of prescriptions and amount prescribed, although not included in the final model, presented indirect impact in the tetracycline resistance probability. From this study, we can infer that tetracycline usage, the time span between last treatment and sampling date, together with herd size and the proportion of animals being treated in a herd, increase the probability of obtaining a resistant isolate.

Space-time clustering of ampicillin resistant Escherichia coli isolated from Danish pigs at slaughter between 1997 and 2005.

In Denmark, antimicrobial resistance in bacteria in animals, animal products and humans, is routinely monitored. This study aimed at determining whether the observed variations in the prevalence of ampicillin resistant Escherichia coli isolated from healthy pigs at slaughter were random or clustered in space and time. Data on E. coli isolates between 1997 and 2005 were obtained from the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) whereas data on the quantity of ampicillin consumed was obtained from the Danish Register of Veterinary Medicines (VetStat). Space-time interaction was assessed using the space-time K-function and detection and location of significant space-time clusters was done using the space-time scan statistic. The space-time K-function analysis provided evidence of space-time interactions in ampicillin resistant E. coli (AREC) isolates in both Funen and Jutland, and Zealand. Significant space-time clusters of resistant E. coli isolates were found in the north eastern part of Jutland and Funen and in the southern part of Zealand. Seasonality was found to have a highly significant effect on space-time clustering in Funen and Jutland. The clusters of ampicillin resistant E. coli appeared at the same time as the national consumption of ampicillin in pigs increased, however antimicrobial consumption at the herd level did not appear to have any effects on space-time clustering in this study. The results could serve as a platform to highlight areas where more investigations on the occurrence and spread of ampicillin resistant E. coli in pig herds should be initiated.