Multiple antigen peptides for specific detection of antibodies to a malaria antigen in human sera.

Multiple antigen peptides (MAP), consisting of a number of peptide copies synthesized on a branching lysyl core, offer a novel approach for rendering small peptides immunoreactive in solid-phase immunoassays. An octameric MAP, carrying 6 repeats of the sequence -N-A-A-G-, tandem repeated in the immunodominant region of the circumsporozoite (CS) protein of Plasmodium malariae, was used as a model to evaluate the suitability of the MAP system in an indirect enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against a parasite antigen in individuals exposed to natural infection. The reaction of endemic sera in ELISA on MAP8-(NAAG)6 was related to that obtained in immunofluorescence on sporozoites, indicating the specificity of the antibody-MAP interaction. The reactivity of immune sera was found to be directed only against the (NAAG)6 moiety of the MAP and not against the lysyl core, since antibody binding to MAP8-(NAAG)6 was completely inhibited by (NAAG)6-NA monomer, but remained uninfluenced when lysyl core was used as competing ligand. The levels of antibodies to MAP8-(NAAG)6, in two groups of individuals naturally exposed to malaria infection, appeared to be related to their respective exposure to the parasite.

In vitro immune recognition of synthetic peptides from the Plasmodium falciparum CS protein by individuals naturally exposed to different sporozoite challenge.

The impact of duration and intensity of sporozoite challenge on the in vitro cell immune response to synthetic peptides of the circumsporozoite (CS) protein of Plasmodium falciparum was investigated in residents of a malaria endemic area in Burkina Faso (West Africa). Lymphocyte proliferation and interferon-gamma (IFN-gamma) production were used to assess immune recognition of synthetic peptides corresponding to the polymorphic Th2R and Th3R regions, to the conserved CS.T3 sequence and to NANP and degenerate NVDP repeats. Immune responses were measured in adults and children from a village where they received more than 100 sporozoite inoculations per year and in adults living in a town, exposed to a 10-100 times lower challenge. A lifetime intense exposure apparently increased the ability to proliferate in response to most peptides in the rural adults, who all produced antibodies to NANP repeats. Surprisingly, cell cultures from these subjects seldom contained appreciable levels of IFN-gamma. In the urban adults, possibly due to the moderate challenge they are exposed to, significant differences in the proliferative potentials of the peptides could be detected. The highest stimulation indices were obtained with the genetically unrestricted CS.T3 peptide. Remarkably, proliferative responses to Th2R and Th3R appeared to be correlated with the humoral response to the CS protein, indicating a T helper significance of the epitopes. The differing proliferative potential of the polymorphic epitopes in the urban adults suggests that polymorphism might delay the development of immune responsiveness under conditions of sporadic transmission. The children from the highly malarious village displayed the lowest proliferative scores, accompanied by a high prevalence of antibodies to NANP repeats. On the basis of these findings, the hypothesis is proposed that a pure B cell reactivity to NANP repeats could ontogenetically precede the mounting of a conventional T-B cooperative immune response.

Vibrio cholerae in the horn of Africa: epidemiology, plasmids, tetracycline resistance gene amplification, and comparison between O1 and non-O1 strains.

The prevalence of Vibrio cholerae O1 and non-O1 has been investigated in numerous Somali regions of the Horn of Africa from 1983 to 1990. From January 1983 to January 1985 and between December 1986 and December 1990, no strains of V. cholerae O1 and 226 strains (5.3%) of V. cholerae non-O1 were isolated from 4,295 diarrhea cases. During a cholera epidemic in 1985 and 1986, the overall case-fatality rate was 13% and the attack rate was 3-3.5 per 1,000 population. Matched case-control studies identified a waterborne route of transmission. A drug-susceptible Ogawa strain from Ethiopia caused the introduction of the disease into northern Somalia. There were two major resistant derivatives of the original strain, and the one resistant to ampicillin, kanamycin, streptomycin, sulfonamide, and tetracycline (TC) predominated in the spreading disease. In 1986, susceptible Ogawa strains quickly displaced this resistant strain. The two incompatibility group C plasmids responsible for the resistance patterns had complex and scattered differences in their structures. Physical analysis of the plasmid DNA region coding for TC resistance demonstrated its genetic amplification in highly resistant variants of Ogawa strains.

Permethrin-impregnated curtains in malaria control.

The impact of permethrin-impregnated curtains on the incidence of malaria episodes, parasitaemia and splenomegaly was assessed during a 22 month period in 2 groups of children aged 0.5-6 years. One group lived in houses where permethrin-impregnated curtains had been installed, the other group lived in houses without curtains. A significant reduction of incidence of malaria episodes, mean parasite density, parasite prevalence and splenomegaly was consistently observed in the intervention group towards the end of the period of moderate transmission, whereas no clear-cut impact could be demonstrated during the high transmission period. The influence of malaria pressure and community utilization on the protective efficiency of curtains is discussed. Because of their acceptability and the ease of reimpregnation, curtains proved to be a suitable technique for integration into primary health care.

Detecting malaria sporozoites in live, field-collected mosquitoes.

A method is described for identifying malaria-infected mosquitoes, without killing them or hampering their fitness. Individual mosquitoes were induced to salivate on coverslips, and sporozoites, deposited on the glass surface, were visualized by Giemsa staining. Of 21 mosquitoes found to contain sporozoites by salivary gland dissection, 13 had delivered sporozoites on coverslips. A positive correlation was found between the amount of saliva expelled and ejection of sporozoites, indicating that the sensitivity of the method may be increased by improving the probing behaviour of the mosquitoes. The procedure described may be suitable for selecting infected mosquitoes which are able to eject sporozoites during probing. Being applicable to wild Anopheles and to large numbers of mosquitoes, the method lends itself for use in field studies on malaria.

[In vivo chemosensitivity of Plasmodium falciparum to chloroquine in Burkina Faso: development of resistance 1988-1990]. / Surveillance de la chimiosensibilite in vivo de Plasmodium falciparum a la chloroquine au Burkina Faso: evolution de la resistance 1988-1990.

A study of Plasmodium falciparum sensitivity to chloroquine was carried out in 1988 and 1990 in 5 localities, representatives of different climatic areas of Burkina Faso. The 7-day in vivo standard test performed in 1988 showed a total clearance failure of 25%. No significant difference with 1990 data was found, except for an increase of the resistance in the area of Fada N'Gourma, close to the border with Benin, Niger and Togo.

[Utilization of permethrin-impregnated curtains by the inhabitants of a rural community in Burkina Faso]. / Utilisation de rideaux impregnés de perméthrine par les habitants d'une communauté rurale du Burkina Faso.

The degree of utilisation of permethrin-impregnated curtains was assessed in a rural community near Ouagadougou, Burkina Faso. Results showed that in the first half of the night, until 11-11.30 p.m., curtains were only partially used by the community. Indeed by this time around 50% of houses had doors well protected by curtains and, over 35% of the community, children and adults, were staying outside. A very marked pattern of this behavior was found, wrong utilisation of curtains being higher in the warm season and lower in the cold season. On the one hand, this situation decreased the potential action of curtains as a barrier to avoid mosquito-man contact and, on the other hand, facilitated the exposure of community to the risk of outdoor infection. These findings may explain the variable level of efficacy showed by curtains in the prevention of malaria morbidity, which seems to be higher in the period between January and February, the winter season in Burkina Faso. The possible application of impregnated curtains as a community-based vector control method is discussed.

Waiting for the vaccine: sporozoite vaccine research entails important progress in malaria epidemiology.

The research efforts aimed at developing a vaccine against malaria, although failing thus far in their main objective, have produced molecular tools of great utility for epidemiological studies. For example, monoclonal antibodies directed against the repeats of Plasmodium circumsporozoite (CS) protein allowed the 2-site assay for detecting sporozoites in mosquitoes to be established. This immunoassay is advantageous compared with the conventional method of salivary gland dissection and microscopic examination, for it makes the identification of the sporozoite species possible, thanks to species-specific aminoacid sequences of the CS repeats. Other examples of vaccine research-derived tools are synthetic peptides reproducing the repetitive part of the CS protein, which allow antibodies to sporozoites, in individuals exposed to malaria, to be detected. Antibodies to the CS repeats of Plasmodium (Laverania) falciparum proved to be a reliable indicator of the intensity of malaria transmission and, therefore, were suitable for monitoring the impact of malaria control programmes. Finally, a project is outlined that, relying on the application of these tools, will aim at characterizing the transmission of Plasmodium (Plasmodium) malariae and at unveiling the possible relationship among different species thriving in the same distribution area, an issue which may become of relevance in view of the likely introduction of a vaccine directed against a single species.

Tirane, Albania: survey on drinking water quality and facilities.

To develop a realistic model of the situation, a study was carried out in four different socioeconomic and hygienic areas of Tirane, namely in the modern and historical centre as well as in the intermediate and peripheral areas. In each area interviewers from the city's Public Health Directorate, contacted randomly, door-to-door, the residents, submitting a questionnaire and collecting water samples at the same time. Our data show relevant differences regarding distribution and quality of drinking water between the centre and peripheral areas. One third of water samples revealed the presence of microorganisms, whereas one fifth had no residual chlorine. Altogether more than 200,000 people in the peripheral areas of Tirane live under low level hygienic conditions. The recent outbreaks of cholera, poliomyelitis and the hyperendemic hepatitis A disease are the dramatic results of the low quality drinking water.

Feasibility of outpatient surgery in the tropics.

The feasibility of outpatient surgery was investigated at a regional hospital in a Third World country. A series of 73 patients with uncomplicated hernias (inguinal, femoral) and hydroceles were allocated at random to receive standard surgical treatment either as inpatients (group 1) or as outpatients (group 2). Patients living within a 4 km radius of the hospital or who agreed to stay in specially provided quarters nearby were eligible to take part in the study. Patients not able to follow the basic hygiene rules taught by the local staff were excluded. The morbidity rate was 5/36 (13.8%) in group 1 and 8/37 (21.6%) in group 2. No statistically significant differences were demonstrated between the complications that occurred in the two groups.

[Impact of utilization of permethrin impregnated curtains on malaria in a rural zone of high transmission in Burkina Faso]. / Impact de l'utilisation des rideaux imprégnés de perméthrine sur le paludisme dans une zone rurale de haute transmission au Burkina Faso.

The efficacy of permethrin impregnated curtains as a malaria control measure was evaluated in a rice field area nearby Ouagadougou (BF). Two groups of children aged 1-5 years matched for age, sex and malaria exposure, were followed through the rainy season of 1987 for illness and febrile episodes. One group of 118 children lived in houses protected with impregnated curtains, the other in houses without curtains. All children were examined for parasitaemia spleen index packed cell volume (PCV) and antisporozoites antibodies at the beginning and the end of the rainy season. During rainy season no difference could be found in the number of clinical episode between the two groups. A reduction in the prevalence of splenomegaly and parasitaemia and an increase in the PCV was observed during the dry season.

A national survey of the prevalence of chloroquine resistant Plasmodium falciparum in Burkina Faso.

Plasmodium falciparum susceptibility to chloroquine was investigated in 10 areas of Burkina Faso in the rainy seasons in 1990-1991. The 7-days in-vivo test was carried out from August to November on children aged 2-8 years with monospecific P. falciparum infection (asexual parasitaemia > 800 microliters-1 of blood), axillary temperature < 37.5 degrees C, and a negative Bergqvist urine test for 4-aminoquinolines. Among 2190 children screened, 366 were selected. Blood samples were collected on days 0, 2, 4 and 7 by finger-prick, and 100 microscopic fields of thick and thin smears were examined for parasite density and species identification. Chloroquine was given under supervision at the standard dose of 25 mg kg-1 over three days (days 0, 1 and 2) with an observation period of one hour after treatment. Parasitaemia did not clear in 63 cases (17.2%) with a 13.4% RII response and 3.8% RIII response. The results do not seem to indicate a decline in the sensitivity of P. falciparum to chloroquine in Burkina Faso during the past two years.