Expression of TNF-alpha-dependent apoptosis-related genes in the peripheral blood of Malagasy subjects with tuberculosis.

The majority of Mycobacterium tuberculosis (Mtb) infections remain asymptomatic with only up to 10% progressing to clinical tuberculosis. However, the constituents of the effective "protective immunity" against tuberculosis responsible for containing most infections remain unknown. Evaluating gene transcriptional profiles in tuberculosis clinical cohorts is one approach to understanding the spectrum of tuberculosis progression. It is clear that apoptosis plays a role in the control of tuberculosis but the utility of apoptosis-related genes as surrogate markers of protection against tuberculosis has not been well investigated. To characterize potential surrogate markers that could discriminate different phases of the clinical tuberculosis spectrum, we investigated gene expression of several TNF-alpha dependent apoptotic genes (TNFR1, TNFR2, FLICE, FLIPs) by real-time RT-PCR of peripheral blood cells from cohorts of individuals with active tuberculosis or potential exposure to tuberculosis. Newly diagnosed tuberculosis patients (n = 23), their close household contacts (n = 80), and community controls (n = 46) were tested at intervals over a period of up to two years. Latent infection or previous Mtb contact was assessed by ELISPOT and TST and complete blood counts were performed during the follow up. Results showed significant upregulation of FLIPs expression by infected individuals regardless of clinical status at entry to the study. A higher percentage of lymphocytes was found in the infected household contacts that remained healthy. In contrast, in individuals with active TB, a significant upregulation of TNFR2 expression, a significantly higher percentage of monocytes and a significantly decreased lymphocyte count were seen, compared to subjects that remained healthy. Moreover, the household contacts who subsequently developed signs of TB also had a significantly high number of monocytes. These data suggest tuberculosis may be associated with decreased T-cell survival (perhaps due to apoptosis) while inhibition of apoptosis in monocytes could lead to a relative increase in these cells: a situation predicted to favour Mtb.

Variation in gamma interferon responses to different infecting strains of Mycobacterium tuberculosis in acid-fast bacillus smear-positive patients and household contacts in Antananarivo, Madagascar.

The majority of healthy individuals exposed to Mycobacterium tuberculosis will not develop tuberculosis (TB), though many may become latently infected. More precise measurement of the human immune response to M. tuberculosis infection may help us understand this difference and potentially identify those subjects most at risk of developing active disease. Gamma interferon (IFN-gamma) production has been widely used as a proxy marker to study infection and to examine the human immune response to specific M. tuberculosis antigens. It has been suggested that genetically distinct M. tuberculosis strains may invoke different immune responses, although how these differences influence the immune responses and clinical outcome in human tuberculosis is still poorly understood. We therefore evaluated the antigen-specific IFN-gamma production responses in peripheral blood mononuclear cells from two cohorts of subjects recruited in Antananarivo, Madagascar, from 2004 to 2006 and examined the influence of the infecting M. tuberculosis strains on this response. The cohorts were sputum-positive index cases and their household contacts. Clinical strains isolated from the TB patients were typed by spoligotyping. Comparison of the IFN-gamma responses with the spoligotype of the infecting clinical strains showed that "modern" M. tuberculosis strains, like Beijing and Central Asian (CAS) strains, tended to induce lower IFN-gamma responses than "ancient" strains, like East African-Indian (EAI) strains, in index cases and their household contacts. These results suggest that new strains may have evolved to induce a host response different from that of ancient strains. These findings could have important implications in the development of therapeutic and diagnostic strategies.

Seroprevalence of hepatitis C and associated risk factors in urban areas of Antananarivo, Madagascar.

BACKGROUND: The risk factors for the transmission of HCV vary substantially between countries and geographic regions. The overall prevalence in south and east Africa region has been estimated to be 1.6% but limited information about the epidemiology of HCV infection in Madagascar is available METHODS: A cross-sectional survey for hepatitis C antibodies was conducted in 2,169 subjects of the general population of Antananarivo to determine seroprevalence of hepatitis C and associated risk factors. RESULTS: The overall seroprevalence was 1.2% (25/2,169). The prevalence did not differ significantly according to gender but it increased with age (Chi2 tendency test, p < 10-5). The variable history of hospitalization, previous therapeutic injections, dental treatment, intravenous drug use, and abnormal ALT and AST were statistically significantly related with the presence of HCV antibodies. No relationship with past history of blood transfusion was observed. CONCLUSION: HCV prevalence in Madagascar seems to be similar to that in most other east African countries. Age appears to be an important risk factor. Iatrogenic causes of HCV transmission need to be further evaluated because all HCV cases had a history of receiving therapeutic injections and data suggested a cumulative effect in relation with therapeutic injections.

HIV type 1 diversity in the Seychelles.

Subtype determination and drug resistance-associated mutations (DRM) detection were performed on 40 HIV-1 Western blot-positive sera detected, obtained from consecutive patients resident in the Seychelles and consulting the Communicable Disease Control Unit, HIV reference center, in Victoria Hospital (Mahe) from October 2005 to June 2006. Amplification and sequencing of at least two of the partial reverse transcriptase, protease, and partial envelope genes were successful for all strains. All three genes sequences were obtained for 39 strains. A high degree of subtype or circulating recombinant forms (CRF) was observed for these 39 strains: A-A1 (17 cases), C (10 cases), B (8 cases), CRF02_AG (2 cases), D (1 case) and CRF01_AE (1 case). According to the ANRS 2006 DRM list and algorithm, none of the 40 isolates was found to be resistant to any protease or reverse transcriptase inhibitors.

Hepatitis C virus infection and genotypes in Antananarivo, Madagascar.

The prevalence of hepatitis C virus (HCV) genotypes in Madagascar is not well known. Serum samples were obtained from 2,169 individuals selected by random sampling in the population living in Antananarivo city. Using HCV antibody test (Monolisa anti-HCV Plus version 2), 36 (1.7%) of the 2,169 samples were positive. The presence of HCV RNA was determined by using reverse transcription polymerase chain reaction amplifying the 5'-untranslated region (UTR): HCV RNA was detected in 17 of the 36 HCV antibodies positive samples. The genotype was determined using BLAST tool with another 5'-UTR fragment. The phylogenetic analysis of the polymerase (NS5b) and envelope (E1/E2) fragment sequences showed a low level of diversity compared to the high diversity in other African countries: subtype 1b (nine cases, 52.9%) and genotype 2 (eight cases, 47.1%) including subtype 2b (six cases), subtype 2k (one case), and one unclassified subtype. BLAST search with the 5'-UTR fragment sequence of this unclassified subtype identified that strain as subtype 2a.

In vitro activities of 18 antimicrobial agents against Staphylococcus aureus isolates from the Institut Pasteur of Madagascar.

BACKGROUND: Staphylococcus aureus, one of the most frequently isolated pathogens in both hospitals and the community, has been particularly efficient at developing resistance to antimicrobial agents. In developed countries, as methicillin-resistant S. aureus (MRSA) has prevailed and, furthermore, as S. aureus with reduced susceptibility to vancomycin has emerged, the therapeutic options for the treatment of S. aureus infections have become limited. In developing countries and especially African countries very little is known concerning the resistance of S. aureus to antibiotics. In Madagascar no data exist concerning this resistance. OBJECTIVE: To update the current status of antibiotic resistance of S. aureus in Antananarivo, Madagascar. METHODS: Clinical S. aureus isolates were collected from patients at the Institut Pasteur of Madagascar from January 2001 to December 2005. Susceptibility tests with 18 antibiotics were performed by the disk diffusion method. RESULTS: Among a total of 574 isolates, 506 were from community-acquired infections and 68 from nosocomial infections. There was no significant difference in the methicillin resistance rate between community-acquired strains (33 of 506; 6.5%) and nosocomial strains (3 of 68, 4.4%). Many MRSA isolates were resistant to multiple classes of antibiotics. Resistance to tetracyclin, trimethoprim-sulfamethoxazole and erythromycin was more common. Among MRSA isolates resistance rates to rifampicin, fusidic acid, gentamicin and ciprofloxacin were lower than that observed with other drugs easily available in Madagascar. No isolates were resistant to glycopeptides. CONCLUSION: The rate of methicillin-resistant S. aureus is not different between community-acquired and nosocomial infections and is still rather low in Madagascar.

Antimicrobial resistance among uropathogens that cause community-acquired urinary tract infections in Antananarivo, Madagascar.

BACKGROUND: Urinary tract pathogens obtained from patients in Madagascar are becoming increasingly resistant to commonly used antibiotics that are readily available at a low price. This poses a real problem for the treatment of community-acquired urinary tract infections (UTIs) in Madagascar. OBJECTIVES: To obtain data on the pathogens responsible for community-acquired UTIs in Antananarivo and on their susceptibility patterns to the antimicrobial agents that are currently used to treat UTIs. METHODS: We conducted a retrospective study on bacteria isolated from the urine of patients at the Institut Pasteur of Madagascar between January 2004 and April 2006. RESULTS: We isolated 903 pathogens from 673 women and 213 men. The most commonly isolated bacteria were Escherichia coli (607 strains), Klebsiella pneumoniae (87 strains), Staphylococcus aureus (35 strains) and Proteus mirabilis (32 strains). Seventy-seven per cent of Gram-negative bacilli were resistant to amoxicillin, 65.7% were resistant to trimethoprim/sulfamethoxazole and more than 15% were resistant to ciprofloxacin. Strains were rarely resistant to more expensive antibiotics (ceftriaxone 5.9%, fosfomycin 4.6%). Most bacteria showed intermediate susceptibility to nitroxolin. Resistance rates of E. coli to ceftriaxone and gentamicin increased significantly between 2005 and 2006, due to the increase in strains harbouring an extended-spectrum beta-lactamase. Gram-positive bacteria, Streptococcaceae and Staphylococcus spp. were rarely resistant, but 9.5% of streptococci were resistant to penicillin A and 8% of staphylococci were resistant to oxacillin. CONCLUSIONS: The rate of amoxicillin- and trimethoprim/sulfamethoxazole-resistant Enterobacteriaceae implies that another antibiotic should be used for empirical treatment and that there is a need for new generic drugs in developing countries, especially in Madagascar.

Seroprevalence of malaria in inhabitants of the urban zone of Antananarivo, Madagascar.

BACKGROUND: Antananarivo, the capital of Madagascar, is located at an altitude of over 1,200 m. The environment at this altitude is not particularly favourable to malaria transmission, but malaria nonetheless remains a major public health problem. The aim of this study was to evaluate exposure to malaria in the urban population of Antananarivo, by measuring the specific seroprevalence of Plasmodium falciparum. METHODS: Serological studies specific for P. falciparum were carried out with an indirect fluorescent antibody test (IFAT). In a representative population of Antananarivo, 1,059 healthy volunteers were interviewed and serum samples were taken. RESULTS: The seroprevalence of IgG+IgA+IgM was 56.1% and that of IgM was 5.9%. The major risk factor associated with a positive IgG+IgA+IgM IFAT was travel outside Antananarivo, whether in the central highlands or on the coast. The abundance of rice fields in certain urban districts was not associated with a higher seroprevalence. CONCLUSION: Malaria transmission levels are low in Antananarivo, but seroprevalence is high. Humans come into contact with the parasite primarily when travelling outside the city. Further studies are required to identify indigenous risk factors and intra-city variations more clearly.

Use of pre-packaged chloroquine for the home management of presumed malaria in Malagasy children.

OBJECTIVE: The main objective of this study was to assess the quality of home malaria management with pre-packaged chloroquine in two areas in the Moramanga district of Madagascar. The knowledge, attitude and practices of care providers in terms of home treatment options were evaluated and compared. The availability of treatment options by studying retailers and community-based service providers was also investigated. METHODS: A cross-sectional investigation in two communities, in the hamlets and villages located close to carers, retailers, community-based service providers and primary health centres was carried out. RESULTS: Carers in the two districts were equally well aware of the use of pre-packaged chloroquine. Their first response to the onset of fever was to treat children with this antimalarial drug at home. The dose administered and treatment compliance were entirely satisfactory (100%) with pre-packaged chloroquine and rarely satisfactory (1.6% to 4.5%) with non pre-packaged chloroquine. In cases of treatment failure, the carers took patients to health centres. Chloroquine was supplied principally by private pharmacies and travelling salesmen selling unpackaged chloroquine tablets. Non pre-packaged chloroquine was the most common drug used at health centres. The frequency of positive rapid malaria tests (P = 0.01) was significantly higher in children treated with non pre-packaged chloroquine (38%) than in children treated with pre-packaged chloroquine (1.3%). CONCLUSION: Home malaria management should be improved in Madagascar. Efforts should focus on communication, the training of community-based service providers, access to pre-packaged drugs and the gradual withdrawal of pre-packaged chloroquine and its replacement by pre-packaged artemisinin-based combination therapies.

High HIV type 1 subtype diversity and few drug resistance mutations among seropositive people detected during the 2005 second generation HIV surveillance in Madagascar.

Subtype determination and detection of drug resistant-associated mutations (DRM) were performed on 31 HIV-1 Western blot-positive sera during the 2005 second-generation HIV surveillance in Madagascar. Amplification and sequencing of at least one of the partial reverse transcriptase, protease, and partial envelope genes were successful for all strains. All three gene sequences were obtained for 28 strains. A high degree of subtype or circulating recombinant forms (CRF) was observed for these 28 strains: A-A1 (eight cases), CRF02_AG (six cases), B (five cases), C (three cases), CRF06_cpx (three cases), CRF10_CD, BC()CRF, and unique RF (one case each). According to the ANRS September 2005 DRM list and algorithm, no DRM was detected in the reverse transcriptase and only one strain bore three major DRM in the protease M46I, I84V, and L90M leading to resistance to indinavir, saquinavir, nelfinavir, atazanavir/ritonavir, and possibly lopinavir.

Use of pre-packaged chloroquine for the home of presumed malaria in Malagasy children

Objective: The main objective of this study was to assess the quality of home malaria management with pre-packaged chloroquine in two areas in the Moramanga district of Madagascar. The knowledge; attitude and practices of care providers in terms of home treatment options were evaluated and compared. The availability of treatment options by studying retailers and community-based service providers was also investigated. Methods: A cross-sectional investigation in two communities; in the hamlets and villages located close to carers; retailers; community-based service providers and primary health centres was carried out. Results :Carers in the two districts were equally well aware of the use of pre-packaged chloroquine. Their first response to the onset of fever was to treat children with this antimalarial drug at home. The dose administered and treatment compliance were entirely satisfactory (100) with pre-packaged chloroquine and rarely satisfactory (1.6to 4.5) with non pre-packaged chloroquine. In cases of treatment failure; the carers took patients to health centres. Chloroquine was supplied principally by private pharmacies and travelling salesmen selling unpackaged chloroquine tablets. Non pre-packaged chloroquine was the most common drug used at health centres. The frequency of positive rapid malaria tests (P=0.01) was significantly higher in children treated with non pre-packaged chloroquine (38) than in children treated with pre-packaged chloroquine (1.3). Conclusions: Home malaria management should be improved in Madagascar. Efforts should focus on communication; the training of community-based service providers; access to pre-packaged drugs and the gradual withdrawal of pre-packaged chloroquine and its replacement by pre-packaged artemisinin-based combination therapies

Evidence of the presence of Seoul virus in Cambodia.

A study was conducted in agricultural and urban areas in Cambodia to assess the presence of hantaviruses in rodent populations. In 1998, rodents were trapped in two villages and in Phnom Penh city around market places and a rubbish dump. IgG antibodies to Hantaan virus were detected in 54 (8.2%) rodents among 660 tested: 6.4% (13/203) among roof rats (Rattus rattus), 20.9% (39/187) among Norway rats (R. norvegicus), 16.7% (2/12) among unidentified Rattus species and none in 183 Polynesian rats (R. exulans) or in 75 bandicoot rats (Bandicota sp.). The presence of the viral genome was detected by a reverse transcription-PCR amplifying part of the sequence coding for the nucleoprotein in the S segment, in 87% of the seropositive rodents. Thirty-one representative cDNAs were sequenced. Phylogenetic studies of the sequences indicated a close relationship with Seoul virus. However, the Cambodian Seoul virus sequences clustered within two different phylogenetic lineages, one associated with R. rattus and the other with R. norvegicus.