Toxocara seropositivity, atopy and asthma: a study in Cuban schoolchildren.

INTRODUCTION: Evidence suggests that human toxocariasis (HT) could stimulate the onset of allergic diseases such as asthma. More specifically, in subjects having a hypothetical 'atopic genotype', HT could boost preexistent allergy symptoms. We tested the latter hypothesis in Cuba, a country where both asthma and HT are prevalent. MATERIAL AND METHODS: In a group of Cuban school-aged children (n = 958), we investigated the association of Toxocara seropositivity and atopic status with asthma. Toxocara seropositivity was diagnosed with ELISA and atopy by allergen skin prick test. Both physician-diagnosed asthma and current wheeze, as determined by International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire, were considered. Associations were assessed using multivariable logistic regression analyses, with either 'physician-diagnosed asthma' or 'current wheeze' as outcome variable. RESULTS: 40.1% of the children were Toxocara seropositive. Prevalences were 21.7% for current wheeze and 32.7% for physician-diagnosed asthma. The odds of having asthma were almost two times higher in atopic children, but only reached borderline significance (OR=1.90, CI 95%: 0.95-3.80 for physician-diagnosed asthma and OR=1.94, CI 95%: 0.98-3.85 for current wheeze). Toxocara seropositivity and physician-diagnosed asthma were associated (OR=1.51, CI 95%: 1.01-2.26). Moreover, in children without antibodies to Toxocara, being atopic was significantly associated with having physician-diagnosed asthma (OR=2.53, CI 95%: 1.63-3.90), while this association was not present in Toxocara positives (OR=1.38, CI 95%: 0.82-2.37). CONCLUSION: Our data confirm previous observations of higher Toxocara seropositivity rates in asthmatic children. Toxocara seropositivity appeared to abrogate the apparent association between atopy and asthma in Cuban children. Although this observation was limited to physician-diagnosed asthma, it challenges the hypothesis that HT stimulates the onset of allergic diseases such as asthma in atopic individuals.

Frequency of antibodies to Toxocara in Cuban schoolchildren.

OBJECTIVE: The aim of the study was to determine the frequency of antibodies to Toxocara in Cuban schoolchildren. METHODS: The frequency of antibodies to Toxocara canis was assessed with a commercial enzyme-linked immunosorbent assays kit in school-aged children from two municipalities of Cuba. Univariate analysis and a multivariable logistic regression analysis adjusted for age, sex, municipality and co-infection with helminth and/or protozoa were conducted. RESULTS: The percentage of children with antibodies to Toxocara was 38.8% (392/1011; 95% CI = 36.8-42.8). Antibody positivity was significantly associated with gender and co-infections with intestinal parasites, but not with age or municipality. CONCLUSION: Cuban children are highly exposed to the Toxocara parasite, corresponding well with reported environmental contamination with Toxocara parasite eggs and T. canis prevalences in dogs in Cuba. Relevant policy makers and the Cuban population need to be better informed about this preventable infection.

Individual and combined effects of ph and lactic acid concentration on Listeria innocua inactivation: development of a predictive model and assessment of experimental variability.

In food technology, organic acids (e.g., lactic acid, acetic acid, and citric acid) are popular preservatives. The purpose of this study was to separate the individual effects of the influencing factors pH and undissociated lactic acid on Listeria innocua inactivation. Therefore, the inactivation process was investigated under controlled, initial conditions of pH (pH0) and undissociated lactic acid ([LaH]0). The resulting inactivation curves consisted of a (sometimes negligible) shoulder period followed by a descent phase. In a few cases, a tailing phase was observed. Depending on the conditions, the descent phase contained one or two log-linear parts or had a convex or concave shape. In addition, the inactivation process was characterized by a certain variability, dependent on the severity of the conditions. Furthermore, in the neighborhood of the growth/no growth interface sometimes contradictory observations occurred. Overall, the individual effects of the influencing factors pH and undissociated lactic acid could clearly be distinguished and were also apparent based on fluorescence microscopy. Appropriate model types were developed and enabled prediction of which conditions of pH0 and [LaH]0 are necessary to obtain a predetermined inactivation (number of decimal reductions) within a predetermined time range.

Modelling Yersinia enterocolitica inactivation in coculture experiments with Lactobacillus sakei as based on pH and lactic acid profiles.

In food processing and preservation technology, models describing microbial proliferation in food products are a helpful tool to predict the microbial food safety and shelf life. In general, the available models consider microorganisms in pure culture. Thus, microbial interactions are ignored, which may lead to a discrepancy between model predictions and the actual microbial evolution, particularly for fermented and minimally processed food products in which a background flora is often present. In this study, the lactic acid mediated negative microbial interaction between the lactic acid bacterium Lactobacillus sakei and the psychrotrophic food pathogen Yersinia enterocolitica was examined. A model describing the lactic acid induced inhibition (i.e., early induction of the stationary phase) of the pathogen [Vereecken, K.M., Devlieghere, F., Bockstaele, A., Debevere, J., Van Impe, J.F., 2003. A model for lactic acid induced inhibition of Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei. Food Microbiology 20, 701-713.] was extended to describe the subsequent inactivation (i.e., decrease of the cell concentration to values below the detection limit). In the development of a suitable model structure to describe the inactivation process, critical points in the variation of the specific evolution rate mu [1/h] with the dynamic (time-varying) pH and undissociated lactic acid profiles were taken into account. Thus, biological knowledge, namely, both pH and undissociated lactic acid have an influence on the microbial evolution, was incorporated. The extended model was carefully validated on new data. As a result, the newly developed model is able to accurately predict the growth, inhibition and subsequent inactivation of Y. enterocolitica in coculture as based on the dynamic pH and lactic acid profiles of the medium.

Mother-to-child transmission of different HIV-1 subtypes among ARV Naïve infected pregnant women in Nigeria.

The rate of mother-to-child transmission (MTCT) of HIV as well as the implications of the circulating multiple subtypes to MTCT in Nigeria are not known. This study was therefore undertaken to determine the differential rates of MTCT of HIV-1 subtypes detected among infected pregnant women before ARV intervention therapy became available in Nigeria. Twenty of the HIV-positive women who signed the informed consent form during pregnancy brought their babies for follow-up testing at age 18-24 months. Plasma samples from both mother and baby were tested for HIV antibody at the Department of Virology, UCH, Ibadan, Nigeria. All positive samples (plasma and peripheral blood mononuclear cells-PBMCs) were shipped to the Institute of Tropical Medicine, Antwerp, Belgium, where the subtype of the infecting virus was determined using the HMA technique. Overall, a mother-to-child HIV transmission rate of 45% was found in this cohort. Specifically, 36.4%, 66.7% and 100% of the women infected with HIV-1 CRF02 (IbNg), G and B, respectively, transmitted the virus to their babies. As far as it can be ascertained, this is the first report on the rate of MTCT of HIV in Nigeria. The findings reported in this paper will form a useful reference for assessment of currently available therapeutic intervention of MTCT in the country.

Mother-to-child transmission of different HIV-1 subtypes among ARV Naïve infected pregnant women in Nigeria

A taxa de transmissão materno-fetal (MTCT) do HIV bem como as implicações dos múltiplos subtipos circulantes para MTCT na Nigéria não são conhecidos. Este estudo foi realizado para determinar as diferentes taxas de MTCT dos subtipos de HIV-1 detectados entre gestantes infectadas antes que a administração da terapia ARV estivesse disponível na Nigéria. Vinte das mulheres HIV positivas que assinaram o consentimento durante a gravidez trouxeram seus filhos para seguimento na idade de 18-24 meses. Amostras de plasma de ambos, mãe e filho foram testadas para anticorpos HIV no Departamento de Virologia, UCH, Ibadan, Nigéria. Todas as amostras positivas (plasma e células mononucleares do sangue periférico - PBMCs) foram enviadas para o Instituto de Medicina Tropical da Antuérpia, Bélgica, onde os subtipos de vírus infectantes foram determinados utilizando-se a técnica HMA. No conjunto, uma taxa de transmissão de HIV, materno-fetal, de 45% foi encontrada neste grupo. Especificamente, 36,4%, 66,7% e 100% das mulheres infectadas com HIV-1 CRF02 (IbNg), G e B, respectivamente, transmitiram o vírus para seus filhos. Até onde pode ser verificado, este é o primeiro relato da taxa de MTCT do HIV na Nigéria. Os achados relatados neste trabalho serão uma útil referência para estimar a qualidade das terapêuticas atuais disponíveis para MTCT neste país.

Towards a novel class of predictive microbial growth models.

Food safety and quality are influenced by the presence (and possible proliferation) of pathogenic and spoilage microorganisms during the life cycle of the product (i.e., from the raw ingredients at the start of the production process until the moment of consumption). In order to simulate and predict microbial evolution in foods, mathematical models are developed in the field of predictive microbiology. In general, microbial growth is a self-limiting process, principally due to either (i) the exhaustion of one of the essential nutrients, and/or (ii) the accumulation of toxic products that inhibit growth. Nowadays, most mathematical models used in predictive microbiology do not explicitly incorporate this basic microbial knowledge. In this paper, a novel class of microbial growth models is proposed. In contrast with the currently used logistic type models, e.g., the model of Baranyi and Roberts [Baranyi, J., Roberts, T.A., 1994. A dynamic approach to predicting bacterial growth in food. International Journal of Food Microbiology 23, 277-294], the novel model class explicitly incorporates nutrient exhaustion and/or metabolic waste product effects. As such, this novel model prototype constitutes an elementary building block to be extended in a natural way towards, e.g., microbial interactions in co-cultures (mediated by metabolic products) and microbial growth in structured foods (influenced by, e.g., local substrate concentrations). While under certain conditions the mathematical equivalence with classical logistic type models is clear and results in equal fitting capacities and parameter estimation quality (see Poschet et al. [Poschet, F., Vereecken, K.M., Geeraerd, A.H., Nicolai, B.M., Van Impe, J.F., 2004. Analysis of a novel class of predictive microbial growth models and application to co-culture growth. International Journal of Food Microbiology, this issue] for a more elaborated analysis in this respect), the biological interpretability and extendability represent the main added value.

Analysis of a novel class of predictive microbial growth models and application to coculture growth.

In this paper, a novel class of microbial growth models is analysed. In contrast with the currently used logistic type models (e.g., the model of Baranyi and Roberts [Baranyi, J., Roberts, T.A., 1994. A dynamic approach to predicting bacterial growth in food. International Journal of Food Microbiology 23, 277-294]), the novel model class, presented in Van Impe et al. (Van Impe, J.F., Poschet, F., Geeraerd, A.H., Vereecken, K.M., 2004. Towards a novel class of predictive microbial growth models. International Journal of Food Microbiology, this issue), explicitly incorporates nutrient exhaustion and/or metabolic waste product effects inducing stationary phase behaviour. As such, these novel model types can be extended in a natural way towards microbial interactions in cocultures and microbial growth in structured foods. Two illustrative case studies of the novel model types are thoroughly analysed and compared to the widely used model of Baranyi and Roberts. In a first case study, the stationary phase is assumed to be solely resulting from toxic product inhibition and is described as a function of the pH-evolution. In the second case study, substrate exhaustion is the sole cause of the stationary phase. Finally, a more complex case study of a so-called P-model is presented, dealing with a coculture inhibition of Listeria innocua mediated by lactic acid production of Lactococcus lactis.

Effect of chemicals on the microbial evolution in foods.

In contrast with most chemical hazardous compounds, the concentration of food pathogens changes during processing, storage, and meal preparation, making it difficult to estimate the number of microorganisms or the concentration of their toxins at the moment of ingestion by the consumer. These changes are attributed to microbial proliferation, survival, and/or inactivation and must be considered when exposure to a microbial hazard is assessed. The number of microorganisms can also change as a result of physical removal, mixing of food ingredients, partitioning of a food product, or cross-contamination (M. J. Nauta. 2002. Int. J. Food Microbiol. 73:297-304). Predictive microbiology, i.e., relating these microbial evolutionary patterns to environmental conditions, can therefore be considered a useful tool for microbial risk assessment, especially in the exposure assessment step. During the early development of the field (late 1980s and early 1990s), almost all research was focused on the modeling of microbial growth over time and the influence of temperature on this growth. Later, modeling of the influence of other intrinsic and extrinsic parameters garnered attention. Recently, more attention has been given to modeling of the effects of chemicals on microbial inactivation and survival. This article is an overview of different applied strategies for modeling the effect of chemical compounds on microbial populations. Various approaches for modeling chemical growth inhibition, the growth-no growth interface, and microbial inactivation by chemicals are reviewed.

Are the cellular immune responses of children and adults with Schistosoma mansoni infection intrinsically different? Cytokines produced ex vivo in response to antigens and mitogens.

In recently exposed communities, intensity of schistosomiasis infection increases as children age and then drops again in adulthood, indicating that host maturity is an important aspect of resistance to schistosomiasis. We investigated whether the cellular immune response to the parasite was correlated with age in subjects with similar daily patterns of exposure, current intensities of infection and number of years of exposure. The cellular immune response of subjects with either 'low' (under 200 eggs per gram (EPG)) or 'high' (over 400 EPG) intensities of infection was investigated, in a recently established focus where subjects had similar histories of exposure and number of years of experience with Schistosoma mansoni. Subject's whole blood was cultured with adult worm antigen (AWA), a mixture of phytohaemagglutinin (PHA) and lipopolysaccharide (LPS), or left unstimulated, and culture supernatants were tested for IL-4, IL-5, IL-10 and IFN-gamma. Children and adults tended to respond differently to schistosome antigen. The most statistically significant illustration of this was the negative correlation between age and IL-5 produced by samples from people with low intensities of infection cultured with AWA (P < 0.003, P < 0.05 after Bonferroni correction). IL-10 produced by samples cultured with PHA and LPS was also notably lower in children than in adults, although not formally significant after Bonferroni correction. This indicates that it is possible for age, independently of intensity of infection or experience with the parasite, to influence the immune response to schistosomiasis.

Human water contacts patterns in Schistosoma mansoni epidemic foci in northern Senegal change according to age, sex and place of residence, but are not related to intensity of infection.

In an epidemic focus in northern Senegal, adults had lower intensities of infection than adolescents, a phenomenon that could not be attributed to immunity acquired over the previous 10-15 years of exposure to the parasite because all age groups had had the same number of years' experience of the worm. This article considers whether this pattern could have been because of higher levels of exposure to the parasite in younger age groups. Personal contact with infected water was recorded using a questionnaire in Schistosoma mansoni foci not more than 3 years old and in another, 10-year-old focus. Many aspects of contact (e.g. frequency, duration or time of day of contact) may contribute to the number of encounters with infective cercariae (true exposure), so various assumptions regarding the relationship between water contact and true exposure were tested resulting in a range of exposure indices. People reported a mean of 4.4 separate contacts, and spent a median of 57 min per day in water. Patterns of water contact differed depending on the exposure index used, e.g. considering duration, males spent a longer time in water than females (P < 0.001). But using frequency, females had more contacts with water than males in most villages (P < 0.001). Generally, exposure levels dropped as people become aged (P < 0.001) and residents of the older focus were more exposed than residents of other foci (P < 0.002). Intensity of (re)infection was not related to exposure either alone or in models incorporating age, sex and/or village irrespective of the index used. There is therefore evidence that age, sex and place of residence determine exposure but none to suggest that exposure had an influence on the relationship between these factors and intensity of infection. We propose therefore that in this population other factors have principal importance in determining intensity of infection.

Effects of gelatin concentration on the growth parameters of Listeria innocua in a model gel system.

In this work, the growth of Listeria innocua was studied responding to the addition of different concentrations of gelatin (see text) model gel system in a modi_ed Brain Heart Infusion medium at 12 C and an initial pH of 6.2. The global number of viable cells as a function of incubation time and the corresponding pH, lactic acid concentration and glucose concentration were measured. Each set of data was fitted with the growth model of Baranyi and Roberts (1994) to estimate the maximum specific growth rate and the maximum cell concentration. Gelatin had a significant e_ect on the growth rate of Listeria innocua, which reduced as the gelatin concentration increased. A tail was observed after a certain concentration of gelatin indicating that there exists a maximum concentration beyond which no further reduction could be observed. There was, however, within the gelatin concentration range studied, no appreciable effect on the maximum cell concentration. A distinct morphological change of colonies was also observed with increasing gelatin concentration.

Predicting inhibition and inactivation of Yersinia enterocolitica through lactic acid production by Lactobacillus sakei.

In food technology, there is a need for models taking into account the interactions between microorganisms, in order to correctly predict the safety and shelf life of food products. When leaving these interactions out of consideration, a discrepancy between the model prediction and the actual microbial evolution may occur for certain types of food products. In this study, a model describing the inhibition of the pathogenic Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei was extended to describe also the subsequent inactivation of Y. enterocolitica. During the development of a suitable model structure to describe the inactivation process, biological knowledge about this process was incorporated. The extended model was able to predict evolution of Y. enterocolitica in coculture as well as in monoculture.

Cost-effectiveness of analgesia after Caesarean section. A comparison of intrathecal morphine and epidural PCA.

BACKGROUND: Patient-controlled analgesia (PCA) techniques and intrathecal morphine are the most widely used treatments for post-Caesarean section pain. However these methods have not been compared with respect to analgesic quality and cost differences. METHODS: Fifty-three patients scheduled for elective or semi-urgent Caesarean section were randomized to receive for postoperative analgesia either epidural PCA with a mixture containing bupivacaine 0.06% and sufentanil 1 microg x ml(-1) or intrathecal morphine 0.15 mg together with the spinal anaesthetic and to be supplemented with paracetamol and tramadol. Analgesic efficacy, side-effects and costs were calculated during 48 h. RESULTS: VAS pain scores both at rest and during mobilization were lower in the PCA group, more particularly during the second postoperative day. Nausea and vomiting were more frequently registered in the morphine treated patients. PCA treated patients stayed longer in the recovery room but required fewer nurse interventions on the surgical ward. Manpower and drug costs were equal in both groups. The differences in total costs (Euro) amounted to euros 33 and were mainly caused by the more expensive equipment required for epidural PCA. Satisfaction and hospital discharge were similar for both treatments. CONCLUSIONS: It was concluded that epidural PCA induced better pain relief, caused less nausea/vomiting but was more expensive than intrathecal morphine.

Study of HIV type 1 gag/env variability in The Gambia, using a multiplex DNA polymerase chain reaction.

A multiplex DNA PCR assay was developed for the simultaneous first-round amplification of HIV-1 gag and env fragments for the heteroduplex mobility assay (HMA). This assay was compared with the conventional amplification assay, using DNA extracted from PBMC samples from 30 HIV-1-seropositive individuals from The Gambia, who were enrolled between 1992 and 1997. From 27 of 30 (90%) samples both gag and env HMA fragments were amplified simultaneously. In one sample only the gag HMA fragment could be amplified by multiplex DNA PCR, and in two samples amplification was negative for both gag and env HMA in multiplex as well as the mono-DNA PCR. Of the 28 Gambian isolates subtyped by gag/env HMA or by sequencing and phylogenetic analysis, the majority (19 of 28; 68%) were intersubtype recombinant. Fifteen of 28 (53%) samples were circulating recombinant form (CRF) CRF02.AG variants. Two isolates clustering with the previously documented Gambian isolate GM4 (previously described as an env GC recombinant) are classified as gag A/env J recombinants.

Development of a one-tube multiplex reverse transcriptase-polymerase chain reaction assay for the simultaneous amplification of HIV type 1 group M gag and env heteroduplex mobility assay fragments.

The emergence of intersubtype recombinant HIV-1 isolates has made it imperative to analyze different regions of HIV-1 genomes. For this purpose a one-tube multiplex RT-PCR, coamplifying first-round amplicons that allow amplification of gag and env heteroduplex mobility assay (HMA) fragments from different HIV-1 group M isolates, was developed, starting with plasma samples. The multiplex RT-PCR assay is sensitive: 115 of 136 (84.5%) samples were positive for both gag and env, positive amplification of the gag fragment was observed in 130 of 136 (95.6%) samples, while for the env fragment 119 of 136 (87.5%) tested positive. The multiplex RT-PCR in combination with gag and env HMA makes large-scale HIV-1 subtyping fast, simple, and more economical.

Predictive modeling of mixed microbial populations in food products: evaluation of two-species models.

Predictive microbiology is an emerging research domain in which biological and mathematical knowledge is combined to develop models for the prediction of microbial proliferation in foods. To provide accurate predictions, models must incorporate essential factors controlling microbial growth. Current models often take into account environmental conditions such as temperature, pH and water activity. One factor which has not been included in many models is the influence of a background microflora, which brings along microbial interactions. The present research explores the potential of autonomous continuous-time/two-species models to describe mixed population growth in foods. A set of four basic requirements, which a model should satisfy to be of use for this particular application, is specified. Further, a number of models originating from research fields outside predictive microbiology, but all dealing with interacting species, are evaluated with respect to the formulated model requirements by means of both graphical and analytical techniques. The analysis reveals that of the investigated models, the classical Lotka-Volterra model for two species in competition and several extensions of this model fulfill three of the four requirements. However, none of the models is in agreement with all requirements. Moreover, from the analytical approach, it is clear that the development of a model satisfying all requirements, within a framework of two autonomous differential equations, is not straightforward. Therefore, a novel prototype model structure, extending the Lotka-Volterra model with two differential equations describing two additional state variables, is proposed to describe mixed microbial populations in foods.

Simplified strategy for detection of recombinant human immunodeficiency virus type 1 group M isolates by gag/env heteroduplex mobility assay. Study Group on Heterogeneity of HIV Epidemics in African Cities.

We developed a heteroduplex mobility assay in the gag gene (gag HMA) for the identification of group M subtypes A to H. The assay covers the region coding for amino acid 132 of p24 to amino acid 20 of p7 (according to human immunodeficiency virus type 1 [HIV-1] ELI, 460 bp). The gag HMA was compared with sequencing and phylogenetic analysis of an evaluation panel of 79 HIV-1 group M isolates isolated from infected individuals from different geographic regions. Application of gag HMA in combination with env HMA on 252 HIV-1- positive plasma samples from Bénin, Cameroon, Kenya, and Zambia revealed a high prevalence of a variety of intersubtype recombinants in Yaoundé, Cameroon (53.8%); Kisumu, Kenya (26.8%); and Cotonou, Bénin (41%); no recombinants were identified among the samples from Ndola, Zambia. The AG(IbNG) circulating recombinant form, as determined by gag HMA, was found to be the most common intersubtype recombinant in Yaoundé (39.4%) and Cotonou (38.5%). Using a one-tube reverse transcriptase PCR protocol, this gag HMA in combination with env HMA is a useful tool for rapidly monitoring the prevalence of the various genetic subtypes as well as of recombinants of HIV-1. Moreover, this technology can easily be applied in laboratories in developing countries.

A prototype model structure for mixed microbial populations in homogeneous food products.

An important factor which has not been included in many models in the field of predictive microbiology is the influence of a background of microflora in a food product. It is however generally known that the growth of a microorganism as a pure culture can be substantially different from its growth in a mixed culture, due to microbial interactions. Because of the importance of these interactions and the lack of suitable modeling techniques in the field of predictive microbiology to describe them, the potential of models in other research fields-namely ecology-to deal with interactions is explored in previous work of the authors. However, a model structure for microbial growth in food products cannot simply be copied from those elaborated in ecology. The structure of a predictive growth model is indeed typical, primarily due to the explicit modeling of a lag phase. The current paper proposes a prototype model structure for growth of mixed microbial populations in homogeneous food products. The model is able to describe a lag phase and reduces to a classical predictive growth model in the special case of single-species growth.