Search

Background: Listeriosis can cause severe disease, especially in fetuses, neonates, older adults, and persons with certain immunocompromising and chronic conditions. We summarize US population-based surveillance data for invasive listeriosis from 2004 through 2009., Methods: We analyzed Foodborne Diseases Active Surveillance Network (FoodNet) data for patients with Listeria monocytogenes isolated from normally sterile sites. We describe the epidemiology of listeriosis, estimate overall and specific incidence rates, and compare pregnancy-associated and nonpregnancy-associated listeriosis by age and ethnicity., Results: A total of 762 listeriosis cases were identified during the 6-year reporting period, including 126 pregnancy-associated cases (17%), 234 nonpregnancy-associated cases(31%) in patients aged <65 years, and 400 nonpregnancy-associated cases (53%) in patients aged ≥ 65 years. Eighteen percent of all cases were fatal. Meningitis was diagnosed in 44% of neonates. For 2004-2009, the overall annual incidence of listeriosis varied from 0.25 to 0.32 cases per 100,000 population. Among Hispanic women, the crude incidence of pregnancy-associated listeriosis increased from 5.09 to 12.37 cases per 100,000 for the periods of 2004-2006 and 2007-2009, respectively; among non-Hispanic women, pregnancy-associated listeriosis increased from 1.74 to 2.80 cases per 100,000 for the same periods. Incidence rates of nonpregnancy-associated listeriosis in patients aged ≥ 65 years were 4-5 times greater than overall rates annually., Conclusions: Overall listeriosis incidence did not change significantly from 2004 through 2009. Further targeted prevention is needed, including food safety education and messaging (eg, avoiding Mexican-style cheese during pregnancy). Effective prevention among pregnant women, especially Hispanics, and older adults would substantially affect overall rates.

Background: Quantitative estimates of the relative risk (RR) of listeriosis among higher-risk populations and a nuanced understanding of the age-specific risks are crucial for risk assessments, targeted interventions, and policy decisions., Method: The RR of invasive listeriosis was evaluated by age, pregnancy status, and ethnicity using 2004-2009 data from the Foodborne Diseases Active Surveillance Network (FoodNet). Nonparametric logistic regression was used to characterize changes in risk with age and ethnicity. Adjusted RRs and 95% confidence intervals (CIs) were evaluated using negative binomial generalized linear models., Results: Among non-pregnancy-associated cases, listeriosis incidence rates increased gradually with age (45-59 years: RR, 4.7; 95% CI, 3.3-6.8; >85 years: RR, 53.8; 95% CI, 37.3-78.9; reference: 15-44 years). The RR was significantly higher for Hispanics than for non-Hispanics (RR, 1.8; 95% CI, 1.3-2.5). Among women of reproductive age (15-44 years), pregnant women had a markedly higher listeriosis risk (RR, 114.6; 95% CI, 68.9-205.1) than nonpregnant women. The RR was higher for Hispanic than non-Hispanic women, regardless of pregnancy status, and this increased during the study period (2004-2006: RR, 1.9; 95% CI, 1.0-3.3; 2007-2009: RR, 4.8; 95% CI, 3.1-7.1)., Conclusions: This study quantifies the increases in risk of listeriosis among older persons, pregnant women, and Hispanics in the United States. Additional research is needed to better describe the independent effects of age on risk while accounting for underlying conditions. These estimates are needed both to optimize risk assessment models and to inform targeted interventions and policy decisions.

Hepatitis C virus (HCV) isolates circulating in Morocco are poorly documented. To determine the subgenotype distribution of HCV in chronically infected patients, serum samples from 185 anti-HCV-positive patients were analyzed. Determination of the HCV genotype and subtype was performed by sequencing the 5'UTR, NS5B and core regions. According to the NS5B phylogeny, the HCV strains primarily belonged to subtypes 1b (75.2%), 2i (19.1%) and 2k (2.8%). Using a Bayesian approach, the mean date of appearance of the most recent common ancestor was estimated to be 1910 for HCV-1b and 1854 for HCV-2i. Although it is currently the most frequent genotype in Morocco and the dominant form in hepatocellular carcinoma, it thus appears that HCV-1b was introduced into the population subsequently to HCV-2i.

Background: The epidemiological and molecular characteristics of hepatitis C virus (HCV) infection in the general population have been poorly investigated in Africa. The aim of this study was to determine the prevalence, genotype distribution and epidemic history of HCV in the Gabonese general population., Methods/principal Findings: A total of 4042 sera collected from adults in 220 villages in all nine administrative areas of the country were screened for antibodies to HCV. HCV NS5B region sequencing was performed for molecular characterization and population genetic analyses. Of 4042 tested sera, 455 (11.2%) were positive. The seroprevalence of HCV varied significantly by administrative area, with the highest rate in Ogooué-Lolo province (20.4%) and the lowest in Ogooué-Maritine province (3.7%). History of parenteral injections, past hospital admission and age over 55 years were independent risk factors for HCV infection (p<0.0001). Phylogenetic analyses showed that 91.9% of the strains were genotype 4 (HCV-4), 5.7% genotype 1 and 2.2% genotype 2. HCV-4 strains were highly heterogeneous, with more than eight subtypes; subtype 4e predominated (57.3%). Coalescence analyses indicated that subtype 4e was the oldest, with an estimated most recent common ancestor of 1702 [95% CI, 1418-1884]. The epidemic profile indicated that it spread exponentially during the first part of the 20th century, probably by iatrogenic transmission., Conclusions/significance: These results confirm the endemicity of HCV subtype 4e in Gabon and show that its spread is due to a cohort effect, with previous, possibly iatrogenic events. More extensive epidemiological studies are needed to better characterize the route of transmission and the dissemination of HCV in Gabon.

Predictive microbiology models are essential tools to model bacterial growth in quantitative microbial risk assessments. Various predictive microbiology models and sets of parameters are available: it is of interest to understand the consequences of the choice of the growth model on the risk assessment outputs. Thus, an exercise was conducted to explore the impact of the use of several published models to predict Listeria monocytogenes growth during food storage in a product that permits growth. Results underline a gap between the most studied factors in predictive microbiology modeling (lag, growth rate) and the most influential parameters on the estimated risk of listeriosis in this scenario (maximum population density, bacterial competition). The mathematical properties of an exponential dose-response model for Listeria accounts for the fact that the mean number of bacteria per serving and, as a consequence, the highest achievable concentrations in the product under study, has a strong influence on the estimated expected number of listeriosis cases in this context., (Published by Elsevier Ltd.)

Background: In southern Cameroon, where SIV(cpz), the source of human immunodeficiency virus 1 (HIV-1) group M, is prevalent among wild chimpanzees, approximately 50% of some human birth cohorts have been infected with hepatitis C virus (HCV) through unclear mechanisms., Methods: To evaluate indirectly the hypothesis that medical interventions contributed to the early emergence of HIV-1, we conducted a cross-sectional study of 451 inhabitants of Ebolowa in southern Cameroon aged 60 years, using HCV as a marker of parenteral transmission of blood-borne viruses. We administered a questionnaire and tested serum for antibodies against HCV. Viral gene sequences were obtained from HCV-positive sera. Molecular clock analyses provided an independent source of information on epidemic history., Results: A total of 252 participants (56%) were HCV seropositive. HCV sequences were amplified and genotyped from 171 individuals. Independent risk factors for HCV seropositivity were older age, having received intravenous treatment against malaria, and having attended an ethnic school (women only), whereas having been circumcised by a traditional practitioner (men only) tended to be associated with HCV. In addition, transfusions were associated with HCV genotype 1 transmission. Molecular clock analyses of HCV genotypes 1, 2, and 4 revealed that each independently underwent exponential growth during the first half of the 20th century., Conclusions: Medical interventions (intravenous antimalarial drugs, transfusions) and to a lesser extent traditional practices (circumcision) were associated with the massive transmission of HCV among this population decades ago. This finding supports the hypothesis that medical interventions contributed to the transmission of blood-borne viruses, perhaps including SIV(cpz) and HIV-1, in the same region during the early 20th century.

Quantitative risk assessment has emerged as a valuable tool to enhance the scientific basis of regulatory decisions in the food safety domain. This article introduces the use of two new computing resources (R packages) specifically developed to help risk assessors in their projects. The first package, "fitdistrplus", gathers tools for choosing and fitting a parametric univariate distribution to a given dataset. The data may be continuous or discrete. Continuous data may be right-, left- or interval-censored as is frequently obtained with analytical methods, with the possibility of various censoring thresholds within the dataset. Bootstrap procedures then allow the assessor to evaluate and model the uncertainty around the parameters and to transfer this information into a quantitative risk assessment model. The second package, "mc2d", helps to build and study two dimensional (or second-order) Monte-Carlo simulations in which the estimation of variability and uncertainty in the risk estimates is separated. This package easily allows the transfer of separated variability and uncertainty along a chain of conditional mathematical and probabilistic models. The usefulness of these packages is illustrated through a risk assessment of hemolytic and uremic syndrome in children linked to the presence of Escherichia coli O157:H7 in ground beef. These R packages are freely available at the Comprehensive R Archive Network (cran.r-project.org)., (Copyright 2010 Elsevier B.V. All rights reserved.)

Background: Multidrug antiretroviral (ARV) regimens including HAART and short-course dual antiretroviral (sc-dARV) regimens were introduced in 2004 to improve Prevention of Mother-to-Child Transmission (PMTCT) in Cameroon. We assessed the effectiveness of these regimens from 6-10 weeks and 12 months of age, respectively., Methodology/findings: We conducted a retrospective cohort study covering the period from October 2004 to March 2008 in a reference hospital in Cameroon. HIV-positive pregnant women with CD4 < or = 350 cells/mm(3) received first-line HAART [regimen 1] while the others received ARV prophylaxis including sc-dARV or single dose nevirapine (sd-NVP). Sc-dARV included at least two drugs according to different gestational ages: zidovudine (ZDV) from 28-32 weeks plus sd-NVP [regimen 2], ZDV and lamuvidine (3TC) from 33-36 weeks plus sd-NVP [regimen 3]. When gestational age was > or = 37 weeks, women received sd-NVP during labour [regimen 4]. Infants received sd-NVP plus ZDV and 3TC for 7 days or 30 days. Early diagnosis (6-10 weeks) was done, using b-DNA and subsequently RT-PCR. We determined early MTCT rate and associated risk factors using logistic regression. The 12-month HIV-free survival was assessed using Cox regression. Among 418 mothers, 335 (80%) received multidrug ARV regimens (1, 2, and 3) and MTCT rate with multidrug regimens was 6.6% [95%CI: 4.3-9.6] at 6 weeks, without any significant difference between regimens. Duration of mother's ARV regimen < 4 weeks [OR = 4.7, 95%CI: 1.3-17.6], mother's CD4 < 350 cells/mm(3) [OR = 6.4, 95%CI: 1.8-22.5] and low birth weight [OR = 4.0, 95%CI: 1.4-11.3] were associated with early MTCT. By 12 months, mixed feeding [HR = 8.7, 95%CI: 3.6-20.6], prematurity [HR = 2.3, 95%CI: 1.2-4.3] and low birth weight were associated with children's risk of progressing to infection or death., Conclusions: Multidrug ARV regimens for PMTCT are feasible and effective in routine reference hospital. Early initiation of ARV during pregnancy and proper obstetrical care are essential to improve PMTCT.

Deli meat was ranked as the highest-risk ready-to-eat food vehicle of Listeria monocytogenes within the 2003 U.S. Food and Drug Administration and U.S. Department of Agriculture, Food Safety and Inspection Service risk assessment. The comparative risk of L. monocytogenes in retail-sliced versus prepackaged deli meats was evaluated with a modified version of this model. Other research has found that retail-sliced deli meats have both higher prevalence and levels of L. monocytogenes than have product sliced and packaged at the manufacturer level. The updated risk assessment model considered slicing location as well as the use of growth inhibitors. The per annum comparative risk ratio for the number of deaths from retail-sliced versus prepackaged deli meats was found to be 4.89, and the per-serving comparative risk ratio was 4.27. There was a significant interaction between the use of growth inhibitors and slicing location. Almost 70% of the estimated deaths occurred from retail-sliced product that did not possess a growth inhibitor. A sensitivity analysis, assessing the effect of the model's consumer storage time and shelf life assumptions, found that even if retail-sliced deli meats were stored for a quarter of the time prepackaged deli meats were stored, retail-sliced product is 1.7 times more likely to result in death from listeriosis. Sensitivity analysis also showed that the shelf life assumption had little effect on the comparative risk ratio.

Home refrigeration temperatures and product storage times are important factors for controlling the growth of Listeria monocytogenes in refrigerated ready-to-eat foods. In 2005, RTI International, in collaboration with Tennessee State University and Kansas State University, conducted a national survey of U.S. adults to characterize consumers' home storage and refrigeration practices for 10 different categories of refrigerated ready-to-eat foods. No distributions of storage time or refrigeration temperature were presented in any of the resulting publications. This study used classical parametric survival modeling to derive parametric distributions from the RTI International storage practices data set. Depending on the food category, variability in product storage times was best modeled using either exponential or Weibull distributions. The shape and scale of the distributions varied greatly depending on the food category. Moreover, the results indicated that consumers tend to keep a product that is packaged by a manufacturer for a longer period of time than a product that is packaged at retail. Refrigeration temperatures were comparable to those previously reported, with the variability in temperatures best fit using a Laplace distribution, as an alternative to the empirical distribution. In contrast to previous research, limited support was found for a correlation between storage time and temperature. The distributions provided in this study can be used to better model consumer behavior in future risk assessments.

The molecular epidemiology of hepatitis C virus (HCV) in the Central African Republic (CAR) is poorly documented. Thus, we conducted phylogenetic analyses of NS5B gene sequences from 58 HCV-infected inhabitants of a remote area of south-west CAR, which indicated that 48 (82.8%) were infected with genotype 4 (HCV-4), five (8.6%) with genotype 2 and five (8.6%) with genotype 1. HCV-4 strains were highly heterogeneous, containing previously described subtypes 4k (48%), 4c (27%), 4r (4%), 4f (4%) and unclassified subtypes (17%). To estimate the epidemic history of these HCV-4 strains, an evolutionary analysis using the coalescent approach was used. The estimated date of the most recent common ancestor of the CAR HCV-4 strains was 1539 (95% confidence intervals, 1317-1697). They exhibited a rapid, exponential spread from 1935 to 1965, simultaneously with what was recently reported in neighbouring Cameroon and Gabon. The hypothesis of a massive iatrogenic transmission during interventions for the control of endemic tropical diseases is discussed.

A model for the assessment of exposure to Listeria monocytogenes from cold-smoked salmon consumption in France was presented in the first of this pair of articles (Pouillot et al., 2007, Risk Analysis, 27:683-700). In the present study, the exposure model output was combined with an internationally accepted hazard characterization model, adapted to the French situation, to assess the risk of invasive listeriosis from cold-smoked salmon consumption in France in a second-order Monte Carlo simulation framework. The annual number of cases of invasive listeriosis due to cold-smoked salmon consumption in France is estimated to be 307, with a very large credible interval ([10; 12,453]), reflecting data uncertainty. This uncertainty is mainly associated with the dose-response model. Despite the significant uncertainty associated with the predictions, this model provides a scientific base for risk managers and food business operators to manage the risk linked to cold-smoked salmon contaminated with L. monocytogenes. Under the modeling assumptions, risk would be efficiently reduced through a decrease in the prevalence of L. monocytogenes or better control of the last steps of the cold chain (shorter and/or colder storage during the consumer step), whereas reduction of the initial contamination levels of the contaminated products and improvement in the first steps of the cold chain do not seem to be promising strategies. An attempt to apply the recent risk-based concept of FSO (food safety objective) on this example underlines the ambiguity in practical implementation of the risk management metrics and the need for further elaboration on these concepts.

It has recently been suggested that HCV genotype 2 (HCV-2) was more recently introduced to Cameroon (Middle Africa) than West African countries. In order to explore the relationships among HCV-2 strains from Cameroon and West Africa, and to estimate the epidemic history of each lineage, a recently-developed Bayesian evolutionary analysis approach was used. The estimated date of the most recent common ancestor (MRCA) of the Cameroon HCV-2 strains, 1630 (95% highest posterior density interval: 1470-1760) was slightly more recent than that of West Africa, 1540 (95% highest posterior density interval: 1380-1680). Estimates of epidemic history indicate significant differences between the two strains. HCV-2 appears to have spread relatively slowly within the West African population from 1630 to 1900, whilst the Cameroon lineages exhibit rapid, exponential spread from 1920 to 1960. This comparative genetic analysis indicates that Cameroon HCV-2 strains are derived from West African strains and that HCV-2 has undergone radically different epidemiological histories in the two regions.

Background: Retention in long-term antiretroviral therapy (ART) program remains a major challenge for effective management of HIV infected people in sub-Saharan Africa. Highly Active Antiretroviral Therapy (ART) discontinuation raises concerns about drug resistance and could negate much of the benefit sought by ART programs., Methods: Based on existing patient records, we assessed determinants of retention in HIV care among HIV patients enrolled in an urban ART at two urban hospitals in Cameroon. Extended Cox regression procedures were used to identify significant predictors of retention in HIV care., Results: Of 455 patients, 314 (69%) were women, median (IQR) age and baseline CD4 cell count were respectively 36 years (30 - 43) and 110 cells/μL (39 - 177). Forty patients (9%) had active tuberculosis (TB) at enrollment. After a median (IQR) follow-up of 18 months (10-18), 346 (75%) were still in care, 8 (2%) were known dead, and 101 (22%) were lost to follow-up (LFU). Severe immunosuppression (CD4 cell count ≤ 50 cells/μL) at baseline (aHR 2.3; 95% CI 1.4 - 3.7) and active tuberculosis upon enrollment (aHR 1.8; 95% CI 1.0 - 3.6) were independent predictors of cohort losses to follow-up within the first 6 months after HAART initiation., Conclusion: These data suggest that three-quarter of HIV patients initiated on HAART remained in care and on HAART by 18 months; however, those with compromised immunologic status at treatment initiation, and those co-infected with TB were at increased risk for being lost to follow-up within the first 6 months on treatment.

This study characterizes salesmen and evaluates the drugs offer and quality of the drugs in the illicit selling network in Yaoundé (Cameroon) and Niamey (Niger). A sample of 75 and 124 drug salesmen working in these cities was questioned using a standardized questionnaire. The prescription of drugs and the advices provided by these poorly trained salesmen could have an important impact in term of public health: 32% and 67% of the salesmen in Yaoundé give systematically or occasionally advices regarding the prescription. The active substances are always present in the 153 drugs of our analysed sample, except for chloroquine-based drugs, among which 5/30 samples did not contain active substance. However, the rate of nonconformity is approximately 50% in the two cities. Complementary studies are needed to explore the origin of these nonconformities, between counterfeiting, low quality of the products provided by the factories, defects of conservation or instability of the formulations.

Background: Buruli ulcer is an infectious disease involving the skin, caused by Mycobacterium ulcerans. This disease is associated with areas where the water is slow-flowing or stagnant. However, the exact mechanism of transmission of the bacillus and the development of the disease through human activities is unknown., Methodology/principal Findings: A case-control study to identify Buruli ulcer risk factors in Cameroon compared case-patients with community-matched controls on one hand and family-matched controls on the other hand. Risk factors identified by the community-matched study (including 163 pairs) were: having a low level of education, swamp wading, wearing short, lower-body clothing while farming, living near a cocoa plantation or woods, using adhesive bandages when hurt, and using mosquito coils. Protective factors were: using bed nets, washing clothes, and using leaves as traditional treatment or rubbing alcohol when hurt. The family-matched study (including 118 pairs) corroborated the significance of education level, use of bed nets, and treatment with leaves., Conclusions/significance: Covering limbs during farming activities is confirmed as a protective factor guarding against Buruli ulcer disease, but newly identified factors including wound treatment and use of bed nets may provide new insight into the unknown mode of transmission of M. ulcerans or the development of the disease.

Background: The Expanded Program on Immunization (EPI) is the most cost-effective measures to control vaccine-preventable diseases. Currently, the EPI schedule is similar for HIV-infected children; the introduction of antiretroviral therapy (ART) should considerably prolong their life expectancy., Methods and Principal Findings: To evaluate the persistence of antibodies to the EPI vaccines in HIV-infected and HIV-exposed uninfected children who previously received these vaccines in routine clinical practice, we conducted a cross-sectional study of children, aged 18 to 36 months, born to HIV-infected mothers and living in Central Africa. We tested blood samples for antibodies to the combined diphtheria, tetanus, and whole-cell pertussis (DTwP), the measles and the oral polio (OPV) vaccines. We enrolled 51 HIV-infected children of whom 33 were receiving ART, and 78 HIV-uninfected children born to HIV-infected women. A lower proportion of HIV-infected children than uninfected children had antibodies to the tested antigens with the exception of the OPV types 1 and 2. This difference was substantial for the measles vaccine (20% of the HIV-infected children and 56% of the HIV-exposed uninfected children, p<0.0001). We observed a high risk of low antibody levels for all EPI vaccines, except OPV types 1 and 2, in HIV-infected children with severe immunodeficiency (CD4(+) T cells <25%)., Conclusions and Significance: Children were examined at a time when their antibody concentrations to EPI vaccines would have still not undergone significant decay. However, we showed that the antibody concentrations were lowered in HIV-infected children. Moreover, antibody concentration after a single dose of the measles vaccine was substantially lower than expected, particularly low in HIV-infected children with low CD4(+) T cell counts. This study supports the need for a second dose of the measles vaccine and for a booster dose of the DTwP and OPV vaccines to maintain the antibody concentrations in HIV-infected and HIV-exposed uninfected children.

Hepatitis C virus (HCV) infection in Cameroon is characterized by widespread seropositivity and great virus genetic diversity (3 genotypes and over 10 subtypes). A total of 244 HCV NS5B sequences of 382-405 bp long (95 type 1, 58 type 2, and 91 type 4) were phylogenetically analyzed to estimate the history of the HCV epidemic in Cameroon. The newly developed Bayesian coalescent approach was used to infer the history of each HCV type. The estimated dates of the most recent common ancestors (MRCA) for genotypes 1 (1500; 95% confidence interval (95% CI): 1300-1650) and 4 (1500; 95% CI: 1350-1700) were in the same range, while the date for genotype 2 MRCA (1600; 95% CI: 1400-1750) was slightly more recent. The mean genetic distance between HCV genotype 1 sequences was greater than that of HCV type 4 sequences, itself greater than that of HCV type 2 sequences. The initial infected populations of all three genotypes did not grow until recently, when they grew exponentially. The growth rate has now begun to slow, with a less steep exponential growth curve. The period of exponential growth of all the three genotypes was between 1920 and 1960. These results (i) confirm that HCV genotypes 1 and 4 have produced long-term endemics, (ii) suggest that genotype 2 was introduced into Cameroon more recently, and (iii) indicate that the exponential spread of the three genotypes between 1920 and 1960 coincided with the mass campaign against trypanosomiasis and mass vaccinations in Cameroon.

A quantitative assessment of the exposure to Listeria monocytogenes from cold-smoked salmon (CSS) consumption in France is developed. The general framework is a second-order (or two-dimensional) Monte Carlo simulation, which characterizes the uncertainty and variability of the exposure estimate. The model takes into account the competitive bacterial growth between L. monocytogenes and the background competitive flora from the end of the production line to the consumer phase. An original algorithm is proposed to integrate this growth in conditions of varying temperature. As part of a more general project led by the French Food Safety Agency (Afssa), specific data were acquired and modeled for this quantitative exposure assessment model, particularly time-temperature profiles, prevalence data, and contamination-level data. The sensitivity analysis points out the main influence of the mean temperature in household refrigerators and the prevalence of contaminated CSS on the exposure level. The outputs of this model can be used as inputs for further risk assessment.

We report the isolation of chikungunya virus from a patient during an outbreak of a denguelike syndrome in Cameroon in 2006. The virus was phylogenetically grouped in the Democratic Republic of the Congo cluster, indicating a continuous circulation of a genetically similar chikungunya virus population during 6 years in Central Africa.

Accidental ingestion of natural waters while bathing carries a risk of infection by waterborne protozoa such as Cryptosporidium, Giardia and, possibly, microsporidia. In order to evaluate this risk, we conducted a one-year prospective study of two recreational lakes and three river sites located near Paris, where bathing and boating are frequent. Twenty-litre water samples were collected monthly from each site. Concentrated samples were submitted to immunomagnetic separation followed by immunofluorescence (IMS-IF) for Cryptosporidium and Giardia detection. PCR and PCR restriction fragment length polymorphism (PCR-RFLP) were used for the genetic characterization of Cryptosporidium species on IMS-IF-positive samples. PCR were systematically performed to detect Enterocytozoon bieneusi. Bacteria counts were also determined. IMS-IF revealed low counts of Giardia cysts and Cryptosporidium oocysts in the recreational lakes, with occasional peaks (max. 165 cysts/10 L and 9 oocysts/10 L). By contrast, the river sites were consistently and sometimes heavily contaminated throughout the year. Enterocytozoon bieneusi was found in only two river samples. PCR-RFLP genotyping showed the presence of C. hominis and C. parvum. No correlation was found between the presence or counts of parasites and bacteria, except between the presence of Giardia and high counts of Escherichia coli and enterococci. Based on a previously developed model for quantitative risk assessment of waterborne parasitic infections, we estimated that the mean risk of infection by Cryptosporidium and Giardia associated with swimming was <10(-4) in the recreational lakes, and frequently higher at the river sites.

We investigated an acute foodborne outbreak in Yaounde, Cameroon, following a private party. Plesiomonas shigelloides, which has rarely been reported as the causative agent of foodborne outbreaks, was strongly suspected. The unusual short incubation period suggested the presence of a preformed toxin within the incriminated food.

Conventional tests for antibody to Hepatitis C virus (HCV) and HCV RNA require considerable time before results are available, remain very expensive and are not adapted to many sub-Saharan African countries where HCV is endemic. The aim of this study was to evaluate the accuracy of an algorithm consisting of two HCV rapid tests to diagnose and predict HCV viremia in patients in Cameroon. Three hundred and twenty nine plasma samples were screened by two HCV rapid tests (ImmunoComb II HCV, PBS Orgenics and Hexagon HCV, Human). Previous evaluation of these samples for HCV antibodies (anti-HCV) by conventional third generation ELISA, considered as a reference test, indicated that 168 were anti-HCV negative and 161 positive. Among the 161 anti-HCV positive plasma, 114 (71%) were HCV RNA-positive by RT-PCR assay. The ImmunoComb II HCV test provided the more sensitive detection of anti-HCV (sensitivity: 99.4% with a 95% CI = 96-100%). Surprisingly, the second HCV rapid test, Hexagon HCV, showed a high capacity to identify non-viremic subjects amongst anti-HCV positive cases (93.6% [95% CI: 82-99%]). These results suggest an algorithm using ImmunoComb II HCV as a first test to screen anti-HCV positive subjects, and Hexagon HCV as a second test to discriminate between viremic and non-viremic HCV seropositive subjects.

This article presents a Listeria monocytogenes growth model in milk at the farm bulk tank stage. The main objective was to judge the feasibility and value to risk assessors of introducing a complex model, including a complete thermal model, within a microbial quantitative risk assessment scheme. Predictive microbiology models are used under varying temperature conditions to predict bacterial growth. Input distributions are estimated based on data in the literature, when it is available. If not, reasonable assumptions are made for the considered context. Previously published results based on a Bayesian analysis of growth parameters are used. A Monte Carlo simulation that forecasts bacterial growth is the focus of this study. Three scenarios that take account of the variability and uncertainty of growth parameters are compared. The effect of a sophisticated thermal model taking account of continuous variations in milk temperature was tested by comparison with a simplified model where milk temperature was considered as constant. Limited multiplication of bacteria within the farm bulk tank was modeled. The two principal factors influencing bacterial growth were found to be tank thermostat regulation and bacterial population growth parameters. The dilution phenomenon due to the introduction of new milk was the main factor affecting the final bacterial concentration. The results show that a model that assumes constant environmental conditions at an average temperature should be acceptable for this process. This work may constitute a first step toward exposure assessment for L. monocytogenes in milk. In addition, this partly conceptual work provides guidelines for other risk assessments where continuous variation of a parameter needs to be taken into account.

During the 2001-2002 outbreak in Gabon, we observed that several dogs were highly exposed to Ebola virus by eating infected dead animals. To examine whether these animals became infected with Ebola virus, we sampled 439 dogs and screened them by Ebola virus-specific immunoglobulin (Ig) G assay, antigen detection, and viral polymerase chain reaction amplification. Seven (8.9%) of 79 samples from the 2 main towns, 15 (15.2%) of 99 samples from Mekambo, and 40 (25.2%) of 159 samples from villages in the Ebola virus-epidemic area had detectable Ebola virus-IgG, compared to only 2 (2%) of 102 samples from France. Among dogs from villages with both infected animal carcasses and human cases, seroprevalence was 31.8%. A significant positive direct association existed between seroprevalence and the distances to the Ebola virus-epidemic area. This study suggests that dogs can be infected by Ebola virus and that the putative infection is asymptomatic.

A pragmatic quantitative risk assessment (QRA) of the risks of waterborne Cryptosporidium parvum infection and cryptosporidiosis in immunocompetent and immunodeficient French populations is proposed. The model takes into account French specificities such as the French technique for oocyst enumeration performance and tap water consumption. The proportion of infective oocysts is based on literature review and expert knowledge. The probability of infection for a given number of ingested viable oocysts is modeled using the exponential dose-response model applied on published data from experimental infections in immunocompetent human volunteers challenged with the IOWA strain. Second-order Monte Carlo simulations are used to characterize the uncertainty and variability of the risk estimates. Daily risk of infection and illness for the immunocompetent and the immunodeficient populations are estimated according to the number of oocysts observed in a single storage reservoir water sample. As an example, the mean daily risk of infection in the immunocompetent population is estimated to be 1.08 x 10(-4) (95% confidence interval: [0.20 x 10(-4); 6.83 x 10(-4)]) when five oocysts are observed in a 100 L storage reservoir water sample. Annual risks of infection and disease are estimated from a set of oocyst enumeration results from distributed water samples, assuming a negative binomial distribution of day-to-day contamination variation. The model and various assumptions used in the model are fully explained and discussed. While caveats of this model are well recognized, this pragmatic QRA could represent a useful tool for the French Food Safety Agency (AFSSA) to define recommendations in case of water resource contamination by C. parvum whose infectivity is comparable to the IOWA strain.

The prevalence of Chlamydia trachomatis infection was 3.78% out of 1,277 volunteer students screened by direct fluorescence assay and Cobas Amplicor PCR. The infection was associated with the nonuse or inconsistent use of condoms in women (P = 0.026) and a previous sexually transmitted infection in men (P = 0.023). The most frequent genotypes determined by sequencing the omp1 genes of 25 clinical isolates were E (44%) and F (20%), and some strains harbored mutations, but E genotype strains did not.

Collaborative work was financed by the EU to develop and assess new diagnostic tools that can differentiate between bovine brucellosis and bovine infections due to Yersinia enterocolitica O:9 either in conjunction with, or as an alternative to, the classical serological, bacteriological or allergic skin tests. Sixteen heifers were experimentally infected with Brucella abortus biovar 1 (five heifers), Brucella suis biovar 2 (two heifers), Y. enterocolitica O:9 (six heifers) and Y. enterocolitica O:3 (three heifers). Four heifers, naturally infected with Y. enterocolitica O:9 that presented aspecific brucellosis serological reactions were also included in the experiment. A self-limited infection was induced in cattle by B. suis biovar 2. All the brucellosis serological tests used, i.e. the slow agglutination test (SAW), the Rose Bengal test (RB), the complement fixation test (CFT), indirect and competitive ELISA's, lacked specificity when used to analyze sera from Y. enterocolitica O:9 infected animals. A Yersinia outer membrane proteins (YOPs)-ELISA was also used and although the test is able to detect a Yersinia group infection, it provided no evidence of whether or not there is a possible brucellosis infection when dual infections are present. The brucellergen IFN-gamma test showed a lack of specificity also. The only test that was proven to be specific is the brucellergen skin test. All brucellosis serological tests, except the indirect ELISA, were limited in their ability to detect B. abortus persistently infected animals. Based on these experimental studies, a strategy was implemented as part of the year 2001 Belgian Brucellosis Eradication Program to substantiate the eradication of bovine brucellosis. Epidemiological inquiries have identified risk factors associated with aspecific serological reactions, possible transmission and infection of cattle by B. suis biovar 2 from infected wild boars; and both legal and administrative measures taken by the veterinary services. No cases of bovine brucellosis have been confirmed in Belgium since March 2000., (Copyright 2002 Elsevier Science B.V.)

During late summer and autumn 2000, a West Nile fever outbreak in southern France resulted in 76 equine clinical cases; 21 horses died. We report the results of a large serosurvey of all equines within a 10-km radius of laboratory-confirmed cases. Blood samples were obtained from 5,107 equines, distributed in groups of 1 to 91 animals. West Nile virus immunoglobulin (Ig) G antibodies were found in 8.5% of animals (n=432). Forty-two percent of the IgG-positive animals were also IgM positive. Horses living in small groups were more affected than those in large groups. The results suggest that West Nile virus is not endemic in the affected area, the Camargue; rather, sporadic outbreaks are separated by long silent periods.

When a perfect reference test (i.e. "gold standard") is not available, it is possible to obtain estimates of test sensitivity and specificity using "latent-class" methods. However, there are few widely available software programs that allow implementation of these procedures. We describe the development of a program (implemented in R and S-Plus software) for this purpose that yields maximum-likelihood estimates of sensitivity, specificity and prevalence. We also have implemented an HTML form, which submits data to a web-based interface to R. The programs can incorporate data obtained from several populations, results of multiple tests, and can account for data obtained from a reference population in which the true status (infected or non-infected) of each individual is known exactly. Two estimation methods are used: a Newton-Raphson procedure and an expectation-maximisation (EM) procedure. The estimation methods assume test independence conditional on the infection status of the individuals and constant test accuracy in each population. A goodness-of-fit statistic and the residuals of pairwise correlation coefficients are calculated to check the validity of these assumptions. Two examples are used to illustrate application and limitations of the programs. The programs are available at www.afssa.fr/interne/tags.htm (Europe) or www.epi.ucdavis.edu/diagnostictests/ (USA).

To investigate the epidemiology of false positive serological reactions (FPSR) in bovine brucellosis, 1259 bovines from 20 herds were sampled on three successive occasions during the winter of 1993-1994 in an area where the herd prevalence rate of FPSR was high. Serum samples were examined by classical brucellosis serological tests (Rose Bengal and complement fixation) and faeces were cultured for the presence of Yersinia enterocolitica O:9. Thirty-nine bovines expressed at least one positive serological reaction during the study. In the herds with FPSR during the 1993-1994 annual brucellosis surveillance campaign, the specificity of the brucellosis serological tests varied significantly from December to March (97.0% to 99.1%). Y enterocolitica O:9 was isolated from 42 bovines but only three of them showed a positive serological response during the study. Y enterocolitica O:9 isolation rates also decreased with time. Young animals and animals having demonstrated FPSR in the past had a greater risk of having a FPSR. Older animals, which rarely showed FPSR, could form a reservoir for Y enterocolitica O:9. While isolation of Y enterocolitica O:9 was not linked to presence of FPSR and conversely, the FPSR phenomenon should be considered, either at the herd level or at the individual level. This work reinforces the link, at least partial, between FPSR and infection by Y enterocolitica O:9.

Three experiments were performed in order to assess the diagnostic value of the Brucellin allergic skin test (AST) in a brucellosis false positive serological reactions (FPSR) context. First, 1259 cattle from 20 Brucella-free herds in a FPSR area were tested twice with AST to estimate its specificity. Secondly, AST and serological tests (complement fixation test [CFT], tube agglutination test, dithiothreitol-microagglutination test and ELISA) sensitivities were evaluated on 111 cattle positive to the Rose Bengal test (RBT) belonging to 15 Brucella-infected herds. Thirdly, AST was used in a field trial to discriminate FPSR from true brucellosis reactions. AST specificity in non-vaccinated cattle was very high (99.83%; confidence interval 95% [CI95%]: 99.67-99.96%). Skin thickening 72 h post-injection was significantly higher on vaccinated cattle (1.42 vs 0.15 mm). In this sub-population, AST specificity decreased significantly to 78% (CI95%: 68-87%). Individual sensitivity of AST relative to Rose Bengal test was 64% (CI95%: 54-72%), while all infected herds were AST positive (n = 15). When associated with CFT, it detected 95% (CI95%: 90-98%) of the infected cattle. These results were consistent with the field trial. In a FPSR context, AST was more specific than RBT or CFT. Therefore, this test could be used at herd level as a confirmation test, on cattle non vaccinated against brucellosis.