Molecular identification of Wolbachia from the filarial nematode Mansonella perstans

Wolbachiae are bacterial endosymbionts of insects and many filarial nematodes whose products trigger inflammation in filarial infections. The dependence of the parasites on their endosymbionts has also led to the use of antibiotics directed against the Wolbachiae, therapy that has been demonstrated to have a profound salutary effect on filarial infections. The identification of Wolbachiae in Mansonella species has been conclusively shown for Mansonella ozzardi (Mo), but not for Mansonella perstans (Mp). Using primers known to amplify the 16S ribosomal DNA of other filarial Wolbachiae, an identical 1393bp band was found in all samples tested. Sequence analysis of these samples demonstrated a single consensus sequence for Mp Wolbachia 16S rDNA that was most similar to Wolbachia sequences from other filarial nematodes. When aligned with the only other Mansonella Wolbachia sequence (Mo) there were only 8 nucleotide differences in the 1369bp overlapping sequence. Phylogenetic dendrograms, examining the relationship of the Mp Wolbachia to other Wolbachia 16S rDNA, showed that the Wolbachia tracked almost identically to the 5S rRNA of their parasite host. Wolbachia surface protein (WSP) was also demonstrated in protein extracted from Mp-containing whole blood. In advance of a treatment trial of Mp, a method for the quantitation of Mp Wolbachia was developed and used to demonstrate not only a relationship between microfilarial numbers and Wolbachia copy numbers, but also to demonstrate the effect of antibiotic on ridding Mp of Wolbachia.     

Susceptibilité d’<Emphasis Type="Italic">Anopheles gambiae sensu lato</Emphasis> aux insecticides communément utilisés dans la lutte antivectorielle au Mali

The objective of this work was to monitor the susceptibility of malaria vectors to insecticides in nine sentinel sites of the National Malaria Control Program in Mali. The study was performed during the rainy seasons of 2010 and 2011. WHO bioassays were conducted using F0 and/or F1 from wild collected females. The insecticides used were lambda-cyhalothrin 0.05%, DDT 4%, permethrin 0.75%, deltamethrin 0.05%, bendiocarb 0.1% and fenitrothion 1.0%. Results showed suspicion of resistance to pyrethroids and organochlorine in An. gambiae s.l. at almost all the sites except Yanfolila where the vector was susceptible to lambda-cyhalothrin (98.0%) [CI 95%, 98-99.8] and to DDT (100%). An. gambiae s.l. was susceptible to bendiocarb in five of the sites (Gao, Bougouni, Djenné, Yanfolila, Tombouctou) while there was a suspicion of resistance at the other sites (Kati, Niono, Bandiagara, Kita). Fenitrothion remains efficient except in the rice area of Niono, where there was a suspicion of resistance with a mortality rate of 92% [IC 95% 88.3-94.8]. Thus, it could be used as an alternative insecticide for IRS in Mali. These results show resistance to pyrethroids, the main insecticide family used in public health (and to some extent in agriculture). This could compromise the malaria vector control efforts in Mali where pyrethroids are used for both in bed nets and in IRS.     

Clinical and biological evolution of HIV-1 seroconverters in Abidjan, Côte d'Ivoire, 1997-2000

OBJECTIVE: To describe the clinical and biologic evolution of HIV-1 infection in Africa. METHODS: One hundred four HIV-1-infected individuals were identified prospectively from regular blood donors in Abidjan, C?te d'Ivoire. The date of seroconversion was estimated from results of sequential serologic tests. Biologic and clinical follow-up was performed every 6 months, starting as early as possible after seroconversion. Case management followed national guidelines. RESULTS: The median interval between estimated seroconversion and study inclusion was 9.7 months, and the median window of seroconversion was 2.8 months. At baseline, all but two patients were asymptomatic; the median CD4 + cell count was 527/mm 3 (interquartile range [IR], 395-684), and the median plasma HIV RNA level was 4.6 log 10 copies/ml (IR, 3.8-4.9). The median follow-up was 23.9 months, and 95% of the patients received primary prophylaxis with co-trimoxazole for opportunistic infections. Of the patients, 1 presented with wasting syndrome, 3 developed tuberculosis, and 17 had a Centers for Disease Control and Prevention category B-defining event. The 3-year AIDS-free and symptom-free probabilities were 96.7% (95% confidence interval [CI], 87.0-99.2] and 79.3% (95% CI, 67.5-87.2), respectively. During the first 3 years of follow-up, we observed that the median plasma viral load stabilized at >4 log 10 copies/ml and that the median CD4 + cell count declined by 20 to 25/mm 3 per year. CONCLUSION: These African seroconverters were moderately immunosuppressed. The median HIV RNA level was high and varied very little during the first 3 years, and there were few clinical events.     

Prevalence and perception of schistosomiasis in a periurban school of Bamako in Mali

The prevalence of urinary and intestinal schistosomiasis, the knowledge, attitude and practice (KAP) regarding the disease among both the children and their parents were investigated in Djikoroni para, a Suburban area of Bamako. The prevalence of Schistosoma haematobium and S. mansoni were respectively 69.8% and 8.7%. Compared with the girls, the boys were significantly more infected by S. haematobium (p = 5.10(-3)), but no difference between the sexes was observed for S. mansoni (p = 0.36). Interviews indicated that 88% of children and 91% of adults considered micro-haematuria as a pathology, but did not know anything about intestinal schistosomiasis. The low sensitivity of micro-haematuria (56.8%) and of the criterion "abdominal pains" (66.7%) indicated that they couldn't be relevant diagnosis signs. The symptoms and mode of transmission were generally well understood for S. haematobium but not for S. mansoni. The later parasite, its intermediate host and their interactions were unknown. The main man water contacts are bathing and playing in the Woyowayanko and in the Niger river. Inadequate perception of schistosomiasis contributes to maintain schistosomiasis at a high level of prevalence in Djikoroni.     

The nonreplicating smallpox candidate vaccines defective vaccinia Lister (dVV-L) and modified vaccinia Ankara (MVA) elicit robust long-term protection

Current smallpox vaccines are live vaccinia viruses that replicate in the vaccinee inducing immunity against the deadly disease smallpox. Replication resulting in virus spread within the host, however, is the major cause of severe postvaccinal adverse events. Therefore, attenuated strains such as modified vaccinia Ankara (MVA) or LC16m8 are candidates as next generation vaccines. These strains are usually grown in primary cells in which mass production is difficult and have an unknown protective potential in humans. Proven vaccine strains of defined origin and modern production techniques are therefore desirable. In this study, defective vaccinia virus (dVV) lacking a gene essential for replication (derived from the Lister vaccine in a complementing cell line) was compared with the Wyeth smallpox vaccine strain and with MVA in mouse animal models using cowpox and ectromelia virus challenge. Similar to MVA, prime-boost immunizations with defective vaccinia induced robust long-term immunity, suggesting it as a promising next generation smallpox vaccine.     

Oocyte morphology predicts outcome of intracytoplasmic sperm injection

To examine the influence of cytoplasmic morphology on the success rate of intracytoplasmic sperm injection (ICSI), the morphology of 837 metaphase II oocytes was assessed after cumulus stripping. The main abnormalities detected were excessive granularity, cytoplasmic inclusions such as vacuoles, smooth endoplasmic reticulum clustering and refractile bodies. Microinjection was performed in 538 oocytes with normal cytoplasm, 142 out of 161 with excessive granularity and 112 out of 138 with cytoplasmic inclusions. Very poor oocytes were not injected. No difference was found in fertilization rate. The embryos achieved cleaved normally and a similar number of good quality embryos among the three groups was noted. The outcome of transfer of embryos derived solely from normal oocytes (group A: 72 patients, 183 embryos) was compared with those from oocytes with cytoplasmic abnormalities (group B: 34 patients, 85 embryos). In group A, 17 clinical pregnancies (24% per patient, implantation rate 10%) were established. In group B, only one clinical pregnancy (3% per patient, implantation rate 1%) was established, from the transfer of embryos derived from oocytes with homogeneous granularity of the cytoplasm. No pregnancy resulted following the transfer of embryos from eggs with cytoplasmic inclusions. The difference was statistically significant. The outcome of ICSI is dependent on the quality of the oocytes retrieved. Normal fertilization and early embryo development were achieved in oocytes with abnormal cytoplasm morphology, but the resulting embryos failed to demonstrate the same implantation potential as those derived from oocytes with normal cytoplasm.      

épidémiologie clinique et parasitologique de la leishmaniose cutanée dans cinq villages du Pays Dogon, Mali

The epidemiology of the cutaneous leishmaniasis (CL) with Leishmania major is poorly documented in Mali. Following reports of CL in the tourist areas of the Dogon country (Bandiagara Escarpment), a joint French and Malian bio-clinical team conducted a field study from 16 to 27 January, 2010. The population of 5 villages has been examined by a dermato-infectiologist and cases were selected by visual inspection of skin lesions. Smears and biopsies (from the lesions) and venous blood were obtained from suspected cases of CL. Diagnosis was performed by light microscopy, in vitro cultures, serology and molecular biology. Fifty patients with skin lesions have been examined. Twenty-one have been suspected as CL. At least one sample was obtained from 18 patients. The lesions were predominantly old, more or less scarring and secondary infected. A skin smear was performed for 15 patients, a skin biopsy for 14 patients: smears and cultures were all negative. The PCR (Leishmania spp.) made on 14 biopsies was positive for 12 patients (86%). The low amount of amplified DNA obtained did not allow the sequencing and identification of the species of Leishmania. Western blot (WB) serology was positive in 11 cases out of 12 (92%). This investigation showed the presence of cutaneous leishmaniasis in Bandiagara. A further investigation is required during transmission period (September-October) to confirm the presence of Leishmania major epidemic in Dogon country.     

Obstetric competence among primary healthcare workers in Mali

Objective

To determine individual and contextual factors associated with emergency obstetric and neonatal care (EmONC) competency among primary healthcare staff in Mali.

Methods

Between November 2011 and April 2012, a competency test was administered to 196 healthcare workers in 65 community health centers in Mali. The test was scored from 0 to 100, and differences among 5 areas of EmONC were assessed. A multilevel linear regression model was used to determine individual and contextual factors associated with score.

Results

The mean score was 66.7 (minimum, 15.9; maximum, 97.7). Knowledge was most deficient for postpartum infection and hypertensive complications. Type of health worker, years of experience, number of days absent, and availability of guidelines for management of obstetric complications within the health center were positively associated with test score (P < 0.05). Availability of guidelines was associated with higher competency of physicians, health technicians, and obstetric nurses (P < 0.001), and seemed to influence the competency of healthcare workers with fewer than 10 years of experience in particular.

Conclusion

Guidelines must be developed that will facilitate standardization of the management of postpartum infection and other less common complications for which healthcare workers show low competence. Strategies to increase use of these guidelines will be necessary.