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Biomedical subjects

B J Vennervald

Publications and source records attributed to B J Vennervald.

At least 19 recordsLinked to original sources

Novel real-time PCr for detection of Schistosoma japonicum in stool.

Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

Animals↗

Epidemiology and morbidity of Schistosoma mansoni infection in a fishing community along Lake Albert in Uganda.

Schistosoma mansoni infection, associated morbidity and symptoms were studied in Piida fishing community at Butiaba, along Lake Albert, Uganda, from November 1996 to January 1997. The study revealed that S. mansoni is highly endemic with an overall prevalence of 72%, a mean intensity of 419.4 eggs per gram (epg) faeces (geometric mean for positives only), with 37.8% of males and 33.0% of females excreting over 1000 epg. Prevalence and intensity peaked in the 10-14 year old age group and decreased with increasing age. Females were less heavily infected than males. Differences were also shown between tribes. Diarrhoea and abdominal pain were commonly reported in Piida. However, no clear-cut correlation between intensity of S. mansoni infection and these conditions could be demonstrated, indicating that retrospective questionnaires concerning S. mansoni related-symptomatology are of limited value. Organomegaly, as assessed by ultrasonography, was frequent and hepatomegaly was associated with heavy S. mansoni infection. No correlation was demonstrated between splenomegaly and infection. This study emphasizes that schistosomiasis mansoni is a major public health problem in Piida fishing community and presumably also in many similar fishing communities. These observations call for immediate intervention and can help in planning long-term strategies for sustainable morbidity control.

Adolescent↗

Hepatosplenic morbidity in two neighbouring communities in Uganda with high levels of Schistosoma mansoni infection but very different durations of residence.

Peri-portal fibrosis can be a serious sequelae of Schistosoma mansoni infection. Age or duration of exposure have been identified as important risk factors, but their relative importance cannot be easily separated. Here, we have compared two cohorts, aged 6-50 years and resident for ten years or since birth, from two neighbouring villages (Booma and Bugoigo) on the eastern shore of Lake Albert, Uganda. Parasitological measurements were similar, whereas the prevalence of peri-portal fibrosis was 5-fold higher in Booma. Data from the cohorts were pooled to assess the relative contribution of age and duration of residency on the risk of disease. Amongst adults, duration of residency was the critical risk factor--individuals aged 17-31 years resident for more 22 years had an almost 12-fold increased risk of fibrosis than those resident for less than 15 years. Height-standardised Splenic Vein Diameter (SVD), Portal Vein Diameter (PVD), Para-sternal Liver Length (PLL) and Spleen Length (SL) values were all higher in Booma, and each organometric parameter except PLL increased with the severity of fibrosis. Our results clearly demonstrate that duration of exposure is a critical risk factor for the development of peri-portal fibrosis and its sequelae in adults. This parameter should therefore be a routine measurement during epidemiological surveys of S. mansoni.

Adolescent↗

Enumeration of CD4 and CD8 T-cells in HIV infection in Zimbabwe using a manual immunocytochemical method.

OBJECTIVES: To enumerate CD4 and CD8 T-cells using the simple and cheap immuno-alkaline phosphatase (IA) method and to compare it with flow cytometry (FC); and to study the effects of duration of sample storage on the IA method results. DESIGN: Method comparison study. SETTING: Blair Research Laboratory, Harare, Zimbabwe. SUBJECTS: 41 HIV positive and 11 HIV negative men and women from Harare participating in HIV studies at Blair Research Laboratory, Zimbabwe. MAIN OUTCOME MEASURES: CD4 and CD8 T-cell counts by FC and the IA method. RESULTS: The IA method and FC were highly correlated for CD4 counts (Spearman rs = 0.91), CD4 percentage (rs = 0.84), CD8 count (rs = 0.83), CD8 percentage (rs = 0.96) and CD4/CD8 ratio (rs = 0.89). However, CD4 cell counts and percentage measured by the IA method were (mean difference +/- SE) 133 +/- 24 cells/microL [corrected] and 6.7 +/- 1.1% higher than those measured by the FC method (p < 0.0001) respectively. CD8 counts and percentages by the IA method were lower than those by the FC method (p < 0.01). Accordingly, the IA method gave a higher CD4/CD8 ratio (p < 0.01). IA method CD4 counts < 300/mL best predicted FC CD4 counts < 200/mL while IA CD4% < 25 best predicted FC CD4% < 14%. IA method CD4/CD8 ratio < 0.8 best predicted FC CD4/CD8 ratio < 0.5. Smears stored for up to 18 months gave results similar to fresh smears. CONCLUSION: The IA method correlates well with but gives CD4 counts and percentages that are higher than those determined by FC. On the contrary, the IA method gives CD8 counts and percentage that are lower than FC values. The method is a cheap and reliable alternative to FC and allows storage of samples for extended periods before analysis, making it an appropriate technology for resource poor countries.

Alkaline Phosphatase↗

Morbidity assessment in urinary schistosomiasis infection through ultrasonography and measurement of eosinophil cationic protein (ECP) in urine.

In a Schistosoma haematobium-endemic village in western Madagascar we evaluated ultrasonography and Eosinophil Cationic Protein (ECP) in urine as means to detect the associated urinary tract pathology. 192 individuals were matched according to age and sex, and grouped into infected persons with bladder and, if present, kidney pathology (n = 96); infected persons without pathology (n = 48) and noninfected persons without pathology (n = 48). The median urinary egg count was significantly higher in individuals with ultrasonographically detectable urinary tract pathology (115 eggs/10 ml urine) than in infected persons without (45 eggs/10 ml of urine). At 136 ng/ml, the median ECP level was significantly higher in the 144 infected individuals than in the 48 noninfected persons (0.35 ng/ml). Egg excretion correlated positively with ECP level. The median ECP level was significantly higher in the group with ultrasonographically detectable urinary tract pathology than in the group without (183 ng/ml vs. 67 ng/ml). The results suggest that minor degrees of pathology, particularly at an early stage of infection with S. haematobium, might be overlooked by ultrasonography despite the presence of marked inflammation, as indicated by markedly increased urinary ECP levels in infected individuals without ultrasonographically detectable urinary tract pathology. ECP may therefore provide important information on the evolution of S. haematobium-associated urinary tract morbidity.

Adolescent↗

Quantitative assessment of eosinophiluria in Schistosoma haematobium infections: a new marker of infection and bladder morbidity.

Eosinophiluria, as quantified by measuring eosinophil cationic protein (ECP) in urinary extracts, microhematuria, egg excretion, and ultrasound-detectable bladder pathology were recorded in Schistosoma haematobium-infected Tanzanian school children at a baseline survey and during an 18-month post-treatment follow-up study. Significant correlations were seen between urinary ECP levels, intensity of infection, and bladder pathology. Treatment resulted in a marked reduction in prevalence and intensity of infection, in a delayed and less marked reduction in ECP levels, and in a resolution of pathology. The overall diagnostic efficiency of the ECP test (cut-off value for the ECP > or =5 ng/ml) in relation to infection was comparable with that of egg count and microhematuria, but with a better sensitivity than a single egg count. In relation to bladder pathology, the diagnostic performance of the ECP test (cut-off value for the ECP > or =25 ng/ml) exceeded that of a single egg count. In addition, the ECP was better in discriminating between different grades of bladder pathology. The present study points to the ECP as a useful marker of both S. haematobium infection and of associated bladder morbidity reflecting the inflammatory status of the bladder wall.

Adolescent↗

Human serum albumin and immunoglobulin on Dracunculus medinensis.

Dracunculus medinensis recovered from infected humans were examined for the occurrence and localization of human serum albumin and immunoglobulins. Immunoelectrophoretic examination of homogenates prepared from adult female worms (ADGW) and first stage larvae (LVGW) showed that anti-human albumin antibodies reacted to both stages of the parasite. By direct fluorescence antibody technique, antigens resembling human albumin and human immunoglobulins (isotype IgG) were identified on the surface of adult female worms. The occurrence of host-like compounds on the parasites may be an adaptation for survival of the parasites. A possible interference of these compounds should be considered in attempts to develop methods for immunological diagnosis of D. medinensis infections.

Animals↗

Adult resistance to schistosomiasis mansoni: age-dependence of reinfection remains constant in communities with diverse exposure patterns.

In a fishing community on Lake Albert in Uganda the pattern of intensity of Schistosoma mansoni infection 6 months after treatment with praziquantel was found to be very similar to reinfection patterns seen in previously studied endemic communities: the profile peaks sharply at around the age of 10 years falling away rapidly to much lower levels in adults. This is in stark contrast to the patterns of water contact, which differ greatly between fishing and non-fishing communities. On Lake Albert, adults appear to be more heavily exposed than children. From these observations we conclude that adults are physiologically (perhaps immunologically) more resistant to infection after treatment than children.

Adolescent↗

Parameters associated with Schistosoma haematobium infection before and after chemotherapy in school children from two villages in the coast province of Kenya.

We evaluated the impact of praziquantel therapy (40 mg/kg body weight) on indicators of infection with Schistosoma haematobium by following a cohort of infected children from schools located 12 km apart in the Coast province of Kenya, at 0, 2, 4, 6, 12 and 18 months after treatment. Within this period, measurements of infection parameters pertaining to egg counts and haematuria (micro-, macro- and history) were evaluated at all time points. The initial prevalence of 100% dropped significantly 8 weeks after treatment with a similar trend in the intensity of infection. Microhaematuria followed the same trend as observed for egg counts while macrohaematuria remained low after treatment. Reinfection following successful therapy differed significantly between schools; in one school the children were reinfected immediately while those in the other remained uninfected despite similar starting prevalences, intensities of infection and cure rates. Transmission between the two areas looked homogeneous before treatment but when both groups were treated, contrasting transmission patterns became evident. In a regression model we evaluated factors that might be associated with reinfection, and after allowing for pretreatment infection level, age and sex, area (school) remained a highly significant predictor.

Adolescent↗

Urine circulating soluble egg antigen in relation to egg counts, hematuria, and urinary tract pathology before and after treatment in children infected with Schistosoma haematobium in Kenya.

A cohort of 117 school children infected with Schistosoma haematobium was followed-up after therapy with praziquantel (0, 2, 4, 6, 12, and 18 months) and various infection and morbidity parameters (egg counts, hematuria, soluble egg antigen [SEA] in urine, and ultrasonography-detectable pathology) were quantified. At the onset of the study, 97% of the children were positive for S. haematobium with a geometric mean egg count of 45.7 eggs/10 ml of urine. Eighty-one percent of the children were positive for SEA in urine with a geometric mean SEA concentration of 218.8 ng/ml of urine. Ninety-two percent and 56% of the children were microhematuria positive and macrohematuria positive, respectively. Two months after treatment, all infection and morbidity indicators had significantly decreased. Reinfection after treatment as determined by detection of eggs in urine was observed by four months post-treatment while the other parameters remained low. The clearance of SEA was slower than that of egg counts while pathology resolved at an even slower pace. Levels of SEA and egg output showed similar correlations with ultrasound detectable pathology; these correlations were better than the correlation between hematuria and pathology.

Adolescent↗

Studies on immunodiagnosis of dracunculiasis. II. Search for circulating antigens.

Sera from individuals living in a dracunculiasis endemic area of northern Ghana were examined for circulating Dracunculus medinensis antigens by applying protocols previously developed for detection of circulating antigens in other helminth infections. Antisera from rabbits immunised with homogenized first stage D. medinensis larvae were used for antigen capture and detection in three different forms, namely non-treated, biotinylated and horseradish peroxidase (HRP)-labelled. Three different preparations of human sera were examined, namely non-treated, pre-treated with polyethylene glycol/ethylenediaminetetra-acetic acid (PEG/EDTA) for analysis of precipitated immune complexes, and pre-treated with trichloroacetic acid (TCA) for analysis of isolated glycoproteins. In both SDS-PAGE/Western blotting and ELISA, significant reactivity was observed between non-treated and treated rabbit-antisera on the one hand and non-treated and treated human sera on the other. However, no significant response differences were observed between sera obtained from individuals with dracunculiasis and non-endemic controls. The reasons are analysed and possible explanations presented. The study provided no evidence that D. medinensis-specific circulating antigens, detectable by relatively simple means, occur in infected individuals.

Adolescent↗

The use of a stereological method to estimate the volume of Schistosoma mansoni granulomas: the effect of zinc deficiency.

A quantitative, stereological method to assess the volume of Schistosoma mansoni granulomas is presented and its use demonstrated in a study of zinc-deficient animals. Twenty-six mice were divided into two groups, and given zinc-deficient or control food. After 4 weeks, the mean weight gain and thymus size were lower in the mice on the zinc-deficient diet. Eight mice in each group were then each infected with 60 S. mansoni cercariae and these mice were killed 12 weeks post-infection. There were then more eggs in the livers of the zinc-deficient mice than in those of the infected controls (7400/female worm v. 3300/female worm; P = 0.02), probably because of impaired intestinal egg expulsion and impaired resorption of eggs in the deficient mice. The median proportion of each liver occupied by granulomas was 0.11 among the zinc-deficient mice and 0.09 among the pair-fed, control mice (P = 0.08), and the median liver volumes were 1400 and 1450 microliters (P = 0.49), respectively. The computed median volume of all the granulomas in each liver was 160 microliters in zinc-deficient and 132 microliters in control mice (P = 0.32), while median granuloma volume per egg was 8.6 and 8.7 nl (P = 0.93), respectively. In contrast to conventional methods, modern stereological methods can provide estimates of granuloma volume or granuloma volume fractions without any assumptions about the size, shape, orientation or distribution of the granulomas in the liver.

Animals↗

Detection and quantification of soluble egg antigen in urine of Schistosoma haematobium-infected children from Kenya.

While research on alternative diagnostic and morbidity markers for infection with Schistosoma haematobium has been going on for a long time, egg counts continue to be used as the gold standard, and infection intensity is thought to reflect the severity of the disease. However, this relationship is not always clear and fluctuation in egg output makes it difficult to classify prevalence correctly. The use of circulating adult worm antigen detection as an alternative diagnostic technique has been applied with varying success. However, this is a measure of worm burden and does not reflect the tissue egg load(s). In the present study we have used an assay that detects soluble egg antigen (SEA) in urine of S. haematobium-infected children, and we have evaluated the applicability of the assay as a diagnostic and morbidity indicator. To evaluate this assay, we have studied a group of 470 children from two schools (Tsunguni and Kibaokiche) in the Coast province of Kenya; 84.8% and 77% were egg-positive while the percentage positive as determined by the SEA-ELISA were 78.8% and 76.2% in Tsunguni and Kibaokiche, respectively. In both schools, SEA levels in urine of S. haematobium-infected children significantly correlated with egg counts (Pearson's r=0.73, P < 0.0001) and with hematuria (Spearman's r=0.65, P < 0.0001). In addition, urinary tract pathology as determined by ultrasound significantly correlated with the SEA levels in urine (Spearman's r=0.3, P < 0.001). The SEA-ELISA compared well with microhematuria within egg count classes and with egg counts within hematuria classes.

Adolescent↗

Evolution of Schistosoma haematobium-related pathology over 24 months after treatment with praziquantel among school children in southeastern Tanzania.

Little is known about the dynamics of pathology due to schistosomiasis following treatment. Public health authorities in endemic areas require such information to decide on the timing of treatment and re-treatment schedules. A study to assess the rate of clearance and reappearance of pathologic lesions due to Schistosoma haematobium using ultrasound has now been carried out in two schools in southeastern Tanzania, an area of moderate-to-high transmission. Baseline data collection found urinary tract pathology in 67% of 533 children. Lesions of the bladder were significantly associated with egg positivity and microhematuria. The attributable fraction estimate of major bladder lesions due to S. haematobium was 75%. In a cohort study, 224 infected children were examined by ultrasound and then treated with a standard dose of 40 mg of praziquantel/kg of body weight. They were re-examined at two, four, six, 12, 18, and 24 months after treatment. Before treatment, 76% had pathologic lesions of the urinary tract. The proportion showing lesions decreased sharply during the first months after treatment to 11% at six months. At 24 months, lesions were detected in 57%, and 11% had developed new severe pathology. In 18 cases, pathology was present throughout, and 34 did not show any pathology throughout the study. This study provides the first detailed report on the evolution of urinary tract pathology due to S. haematobium infections at the community level. The results will help in making decisions on treatment and re-treatment schedules and more generally will provide a basis for designing control strategies in areas of moderate-to-high transmission.

Adolescent↗

The impact of zinc supplementation on growth and body composition: a randomized, controlled trial among rural Zimbabwean schoolchildren.

OBJECTIVES: To assess the effect of zinc supplementation on growth and body composition among schoolchildren. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING AND SUBJECTS: 313 rural Zimbabwean schoolchildren (144 boys and 169 girls, 11-17 y). INTERVENTIONS: Supplementation with zinc (30 or 50 mg) or placebo on schooldays for 12 months. Due to drought, a food programme was in operation during the last eight months of the study. VARIABLES: Weight, height, upper arm circumference, triceps skinfold thickness, and weight-for-age, height-for-age, weight-for-height, arm muscle-area-for-age and arm fat-area-for-age Z-scores. RESULTS: Significant effects on weight gain (0.51 vs 0.14 kg, P = 0.01), weight-for-age Z (-0.08 vs -0.14, P = 0.01) and arm muscle area-for-age Z-score (0.10 vs 0.01, P = 0.03) were seen over the first three months, whereas no effects were seen over the full 12 months. CONCLUSIONS: Zinc deficiency impairing lean body mass and weight gain was documented. However, the effect of zinc seen over the first three months vanished during the last nine months when the food programme was in operation. Zinc deficiency may have persisted, but another nutrient may have become growth limiting during the last nine months.

Adolescent↗