Detection of Schistosoma DNA in genital specimens and urine: A comparison between five female African study populations originating from S. haematobium and/or S. mansoni endemic areas.

Female Genital Schistosomiasis (FGS) is a neglected disease affecting millions, however challenging to diagnose. This explorative descriptive study compares Schistosoma real-time PCR analysis of cervico-vaginal lavages (CVL) with corresponding urine and stool samples of 933 women from five different previously described study populations. Sampling included 310 women from an S. mansoni endemic region in Mwanza, Tanzania and 112 women from a nearby S. haematobium endemic region. Findings were compared with samples collected from S. haematobium endemic regions in South Africa from 394 women and from 117 women from Madagascar of which 79 were urine pre-selected microscopy positive cases from highly-endemic communities and 38 were urine microscopy negatives from a low-endemic community. As anticipated, urine and stool microscopy and gynecological investigations varied substantially between study populations; however, the same Schistosoma real-time PCR was performed in one reference laboratory. Schistosoma DNA was detected in 13% (120/933) of the CVL, ranging from 3% in the S. mansoni Tanzanian endemic region to 61% in the pre-selected Malagasy urine microscopy positive cases. Detectable Schistosoma DNA in CVL was associated with Schistosoma DNA in urine but not with microscopic detection of eggs in urine or by cytological examination. This study confirmed real-time PCR for the detection of Schistosoma DNA in gynecological samples to be a valuable diagnostic tool to study the distribution of FGS within schistosomiasis endemic areas.

Epidemiology of methicillin-resistant Staphylococcus aureus lineages in five major African towns: emergence and spread of atypical clones.

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Africa is poorly documented. From January 2007 to March 2008, we collected 86 MRSA isolates from five African towns, one each in Cameroon, Madagascar, Morocco, Niger and Senegal. Although one or two major clones, defined by the sequence type and staphylococcal cassette chromosome mec type, predominated at each site, genetic diversity (ten clones) was relatively limited in view of the large geographical area studied. Most of the isolates (n = 76, 88%) belonged to three major clones, namely ST239/241-III, a well-known pandemic clone (n = 34, 40%), ST88-IV (n = 24, 28%) and ST5-IV (n = 18, 21%). The latter two clones have only been sporadically described in other parts of the world. The spread of community-associated MRSA carrying the Panton-Valentine leukocidin genes is a cause for concern, especially in Dakar and possibly throughout Africa.

Epidemiology of methicillin-susceptible Staphylococcus aureus lineages in five major African towns: high prevalence of Panton-Valentine leukocidin genes.

The epidemiology of methicillin-susceptible Staphylococcus aureus (MSSA) in Africa is poorly documented. From January 2007 to March 2008, 555 S. aureus isolates were collected from five African towns in Cameroon, Madagascar, Morocco, Niger, and Senegal; among these, 456 unique isolates were susceptible to methicillin. Approximately 50% of the MSSA isolates from each different participating centre were randomly selected for further molecular analysis. Of the 228 isolates investigated, 132 (58%) belonged to five major multilocus sequence typing (MLST) clonal complexes (CCs) (CC1, CC15, CC30, CC121 and CC152) that were not related to any successful methicillin-resistant S. aureus (MRSA) clones previously identified in the same study population. The luk-PV genes encoding Panton-Valentine leukocidin (PVL), present in 130 isolates overall (57%), were highly prevalent in isolates from Cameroon, Niger, and Senegal (West and Central Africa). This finding is of major concern, with regard to both a source of severe infections and a potential reservoir for PVL genes. This overrepresentation of PVL in MSSA could lead to the emergence and spread of successful, highly virulent PVL-positive MRSA clones, a phenomenon that has already started in Africa.

Role of contaminated aspiration tubes in nosocomial outbreak of Klebsiella pneumoniae producing SHV-2 and CTX-M-15 extended-spectrum beta-lactamases.

Klebsiella pneumoniae resistant to ceftazidime was isolated from ten neonates hospitalised between February and March 2006 in two Antananarivo hospitals, Madagascar. The main environmental source, for one hospital in particular, was the liquid used to rinse aspiration tubes in the paediatric wards. The risk of contamination from aspiration tubes is very high in the hospitals of Antananarivo since tap water used to rinse the tubes is not regularly changed. Phenotypical (biotyping and antibiotyping) and genotypical (pulsed-field gel electrophoresis) analysis of all the clinical isolates indicated that nine cases were due to a single clone. This clone carried the genes encoding SHV-2 and CTX-M-15 beta-lactamases. This is the first description of an epidemic due to an ESBL-producing member of the family Enterobacteriaceae in Malagasy hospitals.

[Persistence of an endemic circulation of the West Nile virus in Madagascar]. / Persistance d'une circulation endémique du virus West Nile a Madagascar.

The wide geographic distribution of the West Nile virus and the increase in virulence observed since 1994 in the Mediterranean basin, central Europe and North America, with several outbreaks of lethal encephalitis, demonstrate the importance of regular surveillance of the epidemiological data regarding this virus in the world. The Institut Pasteur de Madagascar has shown between 1975 and 1990 that this arbovirus was most abundant in Madagascar, where it had an endemic circulation. There has been no further study since that time. In order to evaluate the level of circulation, the seroprevalence of anti-West Nile antibodies in children that are 15 or less was measured on two different collections of sera. These collections came from population studies realised respectively in the region of Ambositra in the Highlands in 1996 and in the city of Mahajanga on the north west coast in 1999. The seroprevalence were 2.1% and 10.6% respectively, these results indicate that the circulation of this climatic dependent virus is still significant.

[The current epidemiological situation of cysticercosis in Madagascar]. / Situation épidémiologique actuelle de la cysticercose à Madagascar.

Being associated to fecal-oral transmission, cysticercosis is contracted either by auto-infection or by ingestion of food contaminated with eggs from the pork tape worm (Taenia solium). In the stomach, the larvae named cysticercus (Cysticercus cellulosae) hatches from the eggs and invades the host through the mucosa membrane. Human cysticercosis occurs in highly prevalent proportions in many developing countries including Madagascar where hygiene conditions are deplicable. Serology tests applicable to epidemiological surveillance of cysticercosis and associated pathology in the Malagasy population have been developed: an enzyme-linked immunosorbent assay (ELISA) for screening purpose, and an enzyme-linked immunoelectrotransfer blot assay (EITB) for confirmative testing. Two specific bands (13 and 14 kDa) have been identified as significant markers of the cysticercus in an active (vesicle) stage of the infection when cestocidal treatment is strongly indicated. The same bands may on the other hand be absent at early (cyste) as well as late (calcified) stages of the infection. Series of studies, including 4,375 serum samples, have been undertaken from 1994 until 1999 aiming at determinating the cysticercosis sero-prevalence in different provinces of Madagascar. It was confirmed that cysticercosis is highly frequent on the island, and that there exists a marked variation in the prevalence from 7 to 21% between the different provinces: less than 10% in coastal regions (Mahajanga and Toamasina) increasing to 20% in central regions (Ihosy, Ambositra and Mahasolo). It has also been observed that cysticercosis may occur in individuals at any age, and that it is equally distributed in urban as in rural areas. However, it is more frequently detected in women than in men. Madagascar is an endemic country for cysticercosis, which causes major and severe disease with implications in the public health sector. A national control program is, therefore, urgently warranted.

Geographic differences in hepatosplenic complications of schistosomiasis mansoni and explanatory factors of morbidity.

In a study in three neighbouring villages of southern Madagascar, where Schistosoma mansoni is hyperendemic, ultrasound examination using the Niamey protocol showed marked differences in the burden of disease from one village to another. Hepatosplenic schistosomiasis was more frequent in the village with the highest geometric mean egg counts and the earliest onset of infections, demonstrating that the morbidity induced by S. mansoni may vary greatly within a given area. True representativeness of study populations, a keystone of epidemiological studies, is mandatory to obtain a clear picture of a wide area. Ultrasound examinations in a small number of villages, or even a single one, may be a questionable approach. Using logistic regression analysis, the explanatory variables found to be significantly associated with a risk of severe hepatosplenic disease in our study were sex, age, village of residence and S. mansoni egg counts. On the other hand, a concurrent infection with an intestinal helminth seems to reduce the risk of severe hepatosplenic disease. Further studies should assess the role and possible impact of intestinal helminths on S. mansoni associated-morbidity.

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.

Community-based study of genital schistosomiasis in men from Madagascar.

Detection of Schistosoma haematoblum eggs in 43% of semen samples with Increased levels of eosinophil cationic protein suggests that the genital organs of men are frequently affected with schistosomiasis.

PIP: This community-based cross-sectional study was undertaken to assess the frequency of genital schistosomiasis among men in Androvakely, Madagascar, where Schistosoma haematobium is prevalent. Egg excretion and levels of eosinophil cationic protein (ECP) were measured in paired urine and semen samples obtained from 44 males eligible for the study. Findings revealed that the eggs of S. haematobium were detected in 25 urine samples (57%) and in 19 semen samples (43%). The median egg output (range) in urine was 10 eggs/ml (1-950); in semen, 3 eggs/ejaculate (1-19). Median ECP concentration in urine was 2.2 mcg/l; in semen, 109.0 mcg/l. Moreover, the concentration of ECP in urine was positively correlated with the number of eggs excreted in urine, and increased levels of seminal ECP were significantly associated with the presence of eggs in semen. The presence of eggs in 43% of the semen samples indicated that genitals were common sites for oviposition in men with S. haematobium infection. Based on the findings, a similar effect of genital schistosomiasis on HIV shedding in men, with egg deposition in the genital organs, can lead to an inflammatory response, which may then increase the viral load in semen from HIV-positive people.

Reversibility of Schistosoma mansoni-associated morbidity after yearly mass praziquantel therapy: ultrasonographic assessment.

A parasitological, clinical and ultrasonographic longitudinal study was undertaken in 1993 in a focus hyperendemic for Schistosoma mansoni infection in the central highlands of Madagascar. All the inhabitants were systematically treated with praziquantel. A complete examination and treatment were repeated each year. Among the 289 villagers who underwent the complete 3 years' follow up, 65.9% excreted eggs at the initial survey and the mean egg count of infected individuals was 202 eggs/g. In 1996, the prevalence of infection was 19.3% with a mean egg count of 27 eggs/g and, among inhabitants aged > 44 years, only one was found to be infected. The proportion of individuals complaining of bloody stool decreased from 24.9% in 1993 to 8.4% in 1996. Compared to the initial clinical examination, the age-adjusted prevalence of splenomegaly was significantly lower in 1996, but remained high: 62% in the 10-14 years age group and 59% in individuals aged > 24 years. Ultrasonographic examination after 3 years of praziquantel therapy showed a marked decrease of the overall prevalence of schistosomal hepatic fibrosis, from 28% in 1993 to 10.3% in 1996. This improvement had already been achieved during the second year of follow-up for most subjects. Usually, the reversal of morbidity affected individuals classified as stage 1 at the beginning of the study. Stage 3 was not observed in the last 2 surveys. One patient's ascites disappeared during the follow-up, associated with a significant reversal of periportal fibrosis. Our results indicate that repeated praziquantel therapy can lead to improvement of liver morbidity and the prevention of the development of schistosomal hepatic fibrosis, even in an old-established hyperendemic focus.

[Prevalence of uro-nephrologic complications of urinary bilharziasis in hyperendemic focus in Madagascar]. / Prévalence des retentissements uro-néphrologiques de la bilharziose urinaire dans un foyer hyperendémique de Madagascar.

This prospective study was designed to look for and describe urologic and nephrologic consequences of urinary bilharziosis due to schistosoma haematobium in a hyperendemic hotbed in the middle west of Madagascar. Methodology included clinical examination, kidney and bladder ultrasonography, urine dipsticks and creatininemia. Amongst a population of 574 persons aged 5 years ore more, 436 (76%) had bilharziosis ova in the urine (filtration method). From the clinical point of view, 257 patients (58.9%) had microscopic hematuria, 178 (40.8%) had presently an hematuria; 111 patients (25.5%) suffered from dysuria; 18 patients (4.1%) had limb oedema when 3 patients had present oedema (0.7%). Among 436 checked people, 267 (61.2%) had an ultrasonography abnormality. In 252, it was bladder wall abnormalities (57.8%). They were wall irregularities in 182 cases (41.7%); vesico-ureteral reflux in 22 cases (5.3%); ureteral dilatations in 22 cases (5.3%) and pyelocalyceal dilatations in 61 cases (13.9%). Prevalence of proteinuria 75.2% (316 amongst 420 checked people) of whom 5.7% (24 cases) had 5 g/l or more. Hematuria was found in 352 patients (83.8%) of whom 238 (56.7%) had more than 250 erythrocytes per microliter. Prevalence of leucocyturia was 56.7% (238 cases). Creatininemia was measured in 140 people with positive filtration; it was normal in all except two patients. This study highlights the parallel evolution between parasitic infection and uronephrological manifestations of the disease.

[Effect of praziquantel on the uro-nephrologic complications of urinary bilharziasis]. / Effet du praziquantel sur les retentissements uro-néphrologiques de la bilharziose urinaire.

This study shows the effect of praziquantel as a 12 month treatment on the uro-nephrological consequences of Schistosoma haematobium chronic infection. This was done in a hyperendemic setting in the middle west of Madagascar. 435 people with ova in their urine filtration test were followed up with clinical examination, ultrasonography, urinary sticks and creatininemia. The prevalence of macroscopic hematuria decreased significantly from 32.5% (153 patients) to 4.3% (20 patients) (p < 0.05). Other abnormalities decreased but not significantly. The prevalence of proteinuria decreased from 62.3% (271 cases) to 20.2% (88 cases) (p < 0.05%) when microscopic hematuria varied from 72.4% (315 cases) to 31.5% (271 cases). Leukocyturia remained stable from 49% (213 cases) to 47.8% (207 cases). On ultrasonography, the whole abnormalities varied from 54.1% (256 patients) to 16.7% (79 patients). Prevalence of bladder abnormalities decreased from 50% (237 cases) to 16.3% (77 cases) (p < 0.05); prevalence of vesico-ureteral reflux decreased from 5.1% (23 cases) to 0.2% (1 case) (p < 0.05) and that of pyelocaliceal from 14.6% (54 cases) to 2.5% (12 cases). The reference drug, praziquantel has a clear-cut effect on this chronic pathology. Bladder wall abnormalities are particularly interested by this favourable effects.

Serum concentrations of sICAM-1, sE-, sP- and sL-selectins in patients with Schistosoma mansoni infection and association with disease severity.

Increased serum concentrations of soluble intercellular adhesion molecule-1 (sICAM-1, CD54) and of soluble E- (CD62E), but not soluble P- (CD62P) and L- (CD62 L) selectins, were detected in Malagasy patients living in an hyperendemic focus of Schistosoma mansoni. Levels of sICAM-1 remained elevated for several months after treatment with praziquantel. Serum levels of ICAM-1, but not of other markers, were significantly correlated with the disease severity, as indicated by ultrasonographical data, and with some circulating fibrosis markers (at least hyaluronic acid). sICAM-1 level may reflect endothelial inflammatory reactions, probably harmful, in the liver and may be useful for monitoring morbidity evolution in schistosomiasis mansoni.

Clinical findings in female genital schistosomiasis in Madagascar.

To assess the morbidity of S. haematobium infection in women of reproductive age (15-49 years) in the western part of Madagascar, the village of Betalatala with a prevalence of urinary schistosomiasis in women of 75.6% (95% confidence limit 69.3 to 81.9%) was compared with a neighbouring village with similar socio-economic characteristics and a prevalence of 5.0% (95% confidence limit 0 to 11.75%). The women were questioned in Malagasy about obstetrical history and urogynecological symptoms. They were examined gynaecologically, parasitologically and by ultrasonography. Important STDs were excluded by appropriate diagnostics. In Betalatala significantly more women reported a history of spontaneous abortion (P < 0.01), complaints of irregular menstruation (P < 0.001), pelvic pain (<0.05), vaginal discharge (P < 0.0001), dysuria (P < 0.05) and haematuria (P < 0.01) than in the control village. Biopsies were obtained from the cervix of 36 women with macroscopical lesions, and in 12 cases S. haematobium eggs were found by histological sectioning (33.3%). In the control village no eggs were detected in the histological sections of biopsies taken from 14 women. (P < 0.05). Infections with Candida albicans, Trichomonas vaginalis, Gardnerella vaginalis and Treponema pallidum were found in similar frequencies in both villages. In 9.8% of the women in Betalatala abnormalities of the upper reproductive tract were revealed by ultrasonography versus none in the women from the control village (P < 0.05). Echographic abnormalities of the urinary tract were present in 24% and 3% of the women in the study village and in the control village, respectively (P < 0.0001). These findings were accompanied by an elevated frequency of haematuria (55% versus 20%) and proteinuria (70.4% versus 25%) in the study population (P < 0.0001). Our study indicates that S. haematobium infection in women may not only cause symptoms in the urinary tract, but also frequently in the lower and upper reproductive tract.

Schistosoma mansoni-related morbidity on Ukerewe Island, Tanzania: clinical, ultrasonographical and biochemical parameters.

One thousand six hundred and ninety-five inhabitants of 3 rural villages on Ukerewe Island, Lake Victoria, Tanzania, were examined by clinical, parasitological, ultrasonographic and--in part--serological means to evaluate Schistosoma (S.) mansoni-related morbidity on a community level. Villagers frequently complained of typical colitis symptoms (abdominal pain 80.1%, bloody stools 43.1%, diarrhoea 35.1%); haematemesis, on the other hand, was rare (and reports doubtful in most cases). 16.9% of the population had been given praziquantel previously. Overall S. mansoni prevalence was 86.3%, with a median egg output of 176 eggs per gram (e.p.g.) and maximum output of 17,984 e.p.g. Children and adolescents were infected more severely than adults, men more severely than women. Pretreated individuals excreted significantly fewer ova (median 124 vs 192e.p.g., P < 0.001). Hepatomegaly (determined by ultrasonography) was present in 35%, splenomegaly in 80%. Organomegaly was significantly related to egg output. Pretreated persons had lower rates of splenomegaly and left lobe hepatomegaly. Low-degree periportal fibrosis was common, while severe grades of fibrosis (MANAGIL score II and III) were present in about 6%. About 10% had other abnormalities on liver sonography (irregular parenchymal texture and/or shape); these person passed significantly more S. mansoni ova than others. Clear sonographic signs of portal hypertension were seen in 2.1%. Serum procollagen-IV-peptide and gamma-glutamyl-transferase levels were increased in persons with severe periportal fibrosis, irregular liver texture of portofugal collateral vessels. Thus, S. mansoni infection in the western part of Ukerewe Island is frequent and often severe, leading to a high prevalence of gastrointestinal symptoms. Hepatosplenic involvement does occur, although symptomatic cases of portal hypertension were not identified beyond doubt. The overall level of schistosomal morbidity is thus considered intermediate. Serum procollagen-IV-peptide may be a promising marker of schistosomal liver disease. Our data suggest that S. mansoni infection may also be related to diffuse liver parenchyma alterations in this area.

Schistosoma haematobium infection in western Madagascar: morbidity determined by ultrasonography.

To assess the morbidity related to Schistosoma haematobium infection in western Madagascar, an ultrasonographic examination was performed of 574 inhabitants > 5 years old in a village in an old-established endemic area where no prior systematic antischistosomal treatment had been given. The overall prevalence of infection was 75.9% and the geometric mean egg count of positive individuals was 36 eggs/10 mL of urine. Recent haematuria had been experienced by 31.8% of individuals. Echographic abnormalities of the urinary tract were present in 50.5% of individuals: they were more frequent in males. Bladder wall lesions were observed in 93.1% of individuals with ultrasonographic changes, irregularities of the inner surface being the most common finding. Congestive changes were noted in 8.4% of kidneys, but we observed only 4 severe congestions. Bladder lesions and congestive changes in kidneys predominated in youth; their presence and severity were significantly correlated with egg excretion. In 12 inhabitants, grade 1 periportal fibrosis was observed, but no significant association was found with S. haematobium infection. In a control village, where the prevalence of S. haematobium infection was 7%, moderate congestion of kidneys was observed in 2% of examined inhabitants, and bladder changes in 6%, with a significant relationship with S. haematobium infection.

[Use of ultrasonography in Schistosoma mansoni infections: from the past to the present and future perspectives]. / Utilisation de l'échographie dans l'infection à Schistosoma mansoni: du passé au présent, perspectives d'avenir.

Before the sonographic era, clinics or parasitology even autopsy have been the only sources of basic principles on the morbidity due to Schistosoma mansoni. The exploration of inner organs and their pathologies became easier. The organ's size as well as their echogenicity permit grading. Up till now, any classification is not appropriate. Attempts of standardization haven't yet succeeded, thus any comparison among different endemic regions is not possible. A simple method based on qualitative observation of the periportal fibrosis is proposed to help sonographist for the diagnosis of presumption in front of evocative image and to be a guideline for later investigation.

[Ultrasonographic evaluation of Schistosoma mansoni morbidity: comparison of Cairo/WHO and Managil-Hannover classifications]. / Evaluation échographique de la morbidité liée à Schistosoma mansoni: comparaison des classifications Cairo/OMS et Managil-Hannover.

A ultrasonographical survey of morbidity in schistosomiasis mansoni was carried out in two villages of an endemic area of Madagascar. Using the Managil-Hannover classification, the overall prevalence of morbidity in the first village was of 49.4% and of 39.4% in the second one, while modified Cairo/WHO classification found 9.1% and 1.9% respectively. Thus, results given by the two classifications for a same individual are often discordant. This discrepancy is essentially observed in the lower stage of morbidity while advanced cases are generally well-staged by either of these two methods. These differences in the sonographical assessment of morbidity related to hepatosplenic schistosomiasis make it necessary to continue investigations in order to establish a classification that will have the agreement of everybody.