Malaria transfection and transfection vectors

Malaria remains a leading cause of death due to infectious disease. The completion of the Plasmodium falciparum genome sequencing project and release of preliminary proteomics data have significantly increased our understanding of the biology of this organism. Nonetheless, additional tools for functional analysis of this massive amount of information are now indispensable to further understand the basic biology of this parasite. The genetic manipulation of specific genes via the use of plasmid constructs and transfection represents one such tool.     

Malaria transmission and morbidity

Stable malaria endemicity is maintained over a wide range of transmission intensities in sub-Saharan Africa. This paper considers variations in the clinical manifestations and their consequences with differences in transmission intensity. Epidemiological approaches to malarial disease have concentrated on two clinical syndromes, severe malarial anaemia and cerebral malaria. Within an area the mean age of children with severe malarial anaemia is always lower than that of those with cerebral malaria. In areas of higher malaria transmission children, on average, encounter malaria at a younger age and the mean age of clinical cases is lower. Malarial anaemia tends therefore to be relatively more important under high transmission settings and cerebral malaria tends to gain in importance under lower transmission settings. In a number of studies the total load of malaria morbidity, whether measured as none severe malaria in the community or as severe malaria admitted to hospital, is low under stable low transmission conditions but is at its highest under moderate intensities of transmission. Thereafter it reaches a plateau, or even falls, at the highest transmission intensities. It is not known whether the same is true for mortality in communities living under different transmission settings. Possible implications for changes in patterns of morbidity and mortality following interventions which lower malaria transmission are discussed. It is concluded that such interventions should play an important role in integrated malaria control programmes but that these should involve concomitant introduction of other interventions, in order to minimise the possible risks of a reduced effect as the immune response of the population re-equilibrates in the face of reduced challenge.     

Malaria vectors of Papua New Guinea

Understanding malaria transmission in Papua New Guinea (PNG) requires exact knowledge of which Anopheles species are transmitting malaria and is complicated by the cryptic species status of many of these mosquitoes. To identify the malaria vectors in PNG we studied Anopheles specimens from 232 collection localities around human habitation throughout PNG (using CO₂ baited light traps and human bait collections). A total of 22,970 mosquitoes were individually assessed using a Plasmodium sporozoite enzyme-linked immunosorbent assay to identify Plasmodiumfalciparum, Plasmodiumvivax and Plasmodiummalariae circumsporozoite proteins. All mosquitoes were identified to species by morphology and/or PCR. Based on distribution, abundance and their ability to develop sporozoites, we identified five species as major vectors of malaria in PNG. These included: Anophelesfarauti, Anopheleshinesorum (incriminated here, to our knowledge, for the first time), Anophelesfarauti 4, Anopheleskoliensis and Anophelespunctulatus. Anopheleslongirostris and Anophelesbancroftii were also incriminated in this study. Surprisingly, An. longirostris showed a high incidence of infections in some areas. A newly identified taxon within the Punctulatus Group, tentatively called An. farauti 8, was also found positive for circumsporozoite protein. These latter three species, together with Anopheleskarwari and Anophelessubpictus, incriminated in other studies, appear to be only minor vectors, while Anophelesfarauti 6 appears to be the major vector in the highland river valleys (>1500 m above sea level). The nine remaining Anopheles species found in PNG have been little studied and their bionomics are unknown; most appear to be uncommon with limited distribution and their possible role in malaria transmission has yet to be determined.     

Seasonality in elephant dung decay and implications for censusing and population monitoring in south-western Cameroon

Dung decay rates have been used in the past to correct elephant dung counts to estimates of population numbers. These assume that the dung is in a ‘steady state’ where production rates are equal to disappearance rates. The inter‐seasonal variation in decay rates, however, has never been fully investigated for elephant dung. This study, in the Banyang‐Mbo Wildlife Sanctuary in SW Cameroon, investigated whether a single dung decay rate is really appropriate when converting dung counts to population density estimates. A total of 870 fresh elephant dung‐piles were monitored until they had completely decayed, with at least 60 fresh dung piles located each month of the year between July 1994 to June 1995. The elephant dung decay rate obtained was 55.6% to 66.7% lower than those obtained for the rainforests of Gabon. This has implications for using decay rates obtained elsewhere to compute elephant densities from dung counts. Statistically significant differences were found in the mean duration of dung‐piles between different months and different seasons. Environmental variables, with the exception of rainfall, did not aid in predicting the duration of elephant dung‐piles. The model of Plumptre & Harris was used to estimate the standing crop of dung at any particular time of the year. The results show that, if there was no emigration of elephants from the population, then there was only a three‐month period when the dung was in a ‘steady state’. If emigration occurred during the wet season as is thought to occur then there was no period of ‘steady state’.     

Effect of pathogen-resistant vectors on the transmission dynamics of a vector-borne disease

A model is introduced for the transmission dynamics of a vector-borne disease with two vector strains, one wild and one pathogen-resistant; resistance comes at the cost of reduced reproductive fitness. The model, which assumes that vector reproduction can lead to the transmission or loss of resistance (reversion), is analyzed in a particular case with specified forms for the birth and force of infection functions. The vector component can have, in the absence of disease, a coexistence equilibrium where both strains survive. In the case where reversion is possible, this coexistence equilibrium is globally asymptotically stable when it exists. This equilibrium is still present in the full vector-host system, leading to a reduction of the associated reproduction number, thereby making elimination of the disease more feasible. When reversion is not possible, there can exist an additional equilibrium with only resistant vectors.     

Stream degradation,fish abundance and the potential viability of ornamental fisheries in south-western Cameroon

Fifteen sites on nine second- and third-order streams in the Mount Cameroon area, with varying degrees of human disturbance, were sampled during wet and dry seasons over 21 months in 2003–2005 to estimate their potential for sustainable exploitation of ornamental fishes. In total, 35 species of fish representing 22 genera and 14 families were captured. By biotope, fish populations per 20 m averaged 528 ± 318.5 fish (8.0 ± 3.02 species) in runs, 86 ± 60.9 fish (2.4 ± 1.06 species) in riffles and 819 ± 480.0 fish (7.8 ± 3.15 species) in pools. Riffles contained significantly fewer individuals and species than either runs or pools. Shannon's index was similar for runs and pools, averaging 1.646 ± 0.429 and 1.548 ± 0.345, respectively, but differed significantly for riffles, averaging 0.652 ± 0.473. There were no significant differences in number of individual fish, number of species or Shannon's index among stream order or season, reflecting the stability of these ecosystems throughout the year. Regression of number of fish per 20 linear m of stream, number of species and Shannon's index against a subjective scale related to stream degradation was significant. In particular, the polluted Limbe River sites had significantly fewer individual fish, fewer species and a lower Shannon index across all biotopes than other sites. Fishing efficiency of local gears was estimated at 27% for easily captured species and 14% for the more difficult ones, and could be used to make relative estimates of fish abundance as a means of monitoring changes in fish abundance. Although species diversity in south-western Cameroon was high, typical of tropical streams globally, total abundance was low, and therefore it is unlikely that a profitable ornamental fish trade could be sustained on the basis of fish captured exclusively from the wild.     

Malaria vectors in Lake Victoria and adjacent habitats in western Kenya

The prevalence of malaria among the residents of the Lake Victoria basin remains high. The environment associated with the lake may maintain a high number of malaria vectors. Lake habitats including water hyacinths have been suspected to be the source of vectors. This study investigated whether malaria vectors breed in the lake habitats and adjacent backwater pools. Anopheline larvae were collected within the littoral zone of the lake and adjacent pools located along approximately 24.3 km of the lakeshore in western Kenya, and their breeding sites characterized. Three primary vector species, Anopheles arabiensis, Anopheles gambiae s.s. and Anopheles funestus s.s., and three potential vectors, were found in the lake habitats. Unexpectedly, An. arabiensis was the most dominant vector species in the lake sampling sites. Its habitats were uncovered or covered with short grass. A potential secondary malaria vector, Anopheles rivulorum, dominated the water hyacinths in the lake. Most breeding sites in the lake were limited to areas that were surrounded by tall emergent plants, including trees, and those not exposed to waves. Nearly half of adjacent habitats were lagoons that were separated from the lake by sand bars. Lagoons contained a variety of microhabitats. Anopheles arabiensis dominated open habitats, whereas An. funestus s.s. was found mainly in vegetated habitats in lagoons. The current study confirmed that several breeding sites are associated with Lake Victoria. Given that Lake Victoria is the second largest lake in the world, the lake related habitats must be extensive; therefore, making targeted vector control difficult. Further exploration is necessary to estimate the effects of lake associated habitats on malaria transmission so as to inform a rational decision-making process for vector control.     

Environmental consideration in a coastal engineering project the Delta Project in the south-western Netherlands

Summary In 1958 the Delta Plan was laid down in an Act of Parliament. By changing the attitudes the goals are partly radically changed. The Delta Works will finally (1985) result in an area including almost all kinds of aquatic habitats: fresh-, brackish- and saltwater lakes, some eutrophic and some mesotrophic estuaries, with and without a saline gradient. By segregation of the basins by dams and by integrative possibilities with management tools like sluices, canals and even a storm surge barrier, a scale of management possibilities is realized. Choices have to be prepared and executed. Since 1971 the Delta Project is guided by professional environmentalist, who are fully engaged to the organization. In this paper the first experiences are presented. A problem analysis is given of the environmental problems cohering with the project. It concludes with the main environmental problems in this delta. The research strategy describes that there was no planned research in schemes initially. The research was started based on questions arising from all the schemes and finally from a basin analysis. Then the ordening of question took place by the formulation of research projects, which were ordened to themes. In the advice strategy the technique of advice, policy analysis, is discussed. Then the contents of advices are described. This includes the conclusion of shortage of knowledge about ecological relations, the system approach; the formulation of boundary conditions and peoples interest in functionalities. Management guidelines are derived from the boundary conditions, the reappraisals of functions and the positively evaluated internal processes. The main tasks of environmental guidance are described. At last the basin strategy is described. This part starts with the concept physical planning procedures, the need for special treatments for large basins. The concept management is described, as well as the integration of the management plans and physical planning. This part ends with a few proposals.     

Malaria parasite transmission stages: an update

The Molecular Approaches to Malaria 2004 meeting provided an opportunity to see the impressive progress in all research fields and in the four years since the previous Molecular Approaches to Malaria meeting, when much of the Plasmodium falciparum genome sequence was already available. Study of the part of the Plasmodium life cycle associated with transmission through the vector, which begins with the commitment of blood-stage forms to sexual development, has been especially fruitful. This success is a result of several reasons including: (i) the availability of the genome sequence; (ii) the availability of good animal models that allow parasite culture and facile in vivo studies of many of the life cycle stages involved in transmission; (iii) the availability of genetic manipulation technologies for the animal models of malaria, as well as P. falciparum; and (iv) the ability to study lethal gene knockouts at this stage of the life cycle.     

Malaria transmission blocking immunity and sexual stage vaccines for interrupting malaria transmission in Latin America

Malaria is a vector-borne disease that is considered to be one of the most serious public health problems due to its high global mortality and morbidity rates. Although multiple strategies for controlling malaria have been used, many have had limited impact due to the appearance and rapid dissemination of mosquito resistance to insecticides, parasite resistance to multiple antimalarial drug, and the lack of sustainability. Individuals in endemic areas that have been permanently exposed to the parasite develop specific immune responses capable of diminishing parasite burden and the clinical manifestations of the disease, including blocking of parasite transmission to the mosquito vector. This is referred to as transmission blocking (TB) immunity (TBI) and is mediated by specific antibodies and other factors ingested during the blood meal that inhibit parasite development in the mosquito. These antibodies recognize proteins expressed on either gametocytes or parasite stages that develop in the mosquito midgut and are considered to be potential malaria vaccine candidates. Although these candidates, collectively called TB vaccines (TBV), would not directly stop malaria from infecting individuals, but would stop transmission from infected person to non-infected person. Here, we review the progress that has been achieved in TBI studies and the development of TBV and we highlight their potential usefulness in areas of low endemicity such as Latin America.     

Gut microbiota and parasite transmission by insect vectors

In the gut of some insect vectors, parasites ingested with the bloodmeal decrease in number before coming into contact with host tissues. Many factors could be responsible for this reduction in parasite number but the potentially important role of the large communities of naturally occurring microorganisms that exist alongside the newly ingested parasites in the vector midgut has been largely overlooked. Some previous reports exist of the inhibition of parasite development by vector gut microbiota and of the killing of Trypanosoma cruzi and Plasmodium spp. by prodigiosin produced by bacteria. Based on this evidence, we believe that the microbiota present in the midgut of vector insects could have important roles as determinants of parasite survival and development in insect vector hosts and, therefore, contribute to the modulation of vector competence for many important diseases.     

Malaria sporozoites: survival, transmission and disease control

A basic fact in malariology is that human malaria infections result from the bite of an infective anopheline mosquito. Without sporozoite transmission there is no malaria. Here, John Beier highlights our understanding of sporozoite biology from the perspective of how sporozoites develop and survive in mosquitoes, how they are transmitted during bloodfeeding, and how key elements in sporozoite-vector relationships present new opportunities for tackling critical questions relevant for disease control.     

Association of Cytokine and Toll-Like Receptor Gene Polymorphisms with Severe Malaria in Three Regions of Cameroon

P. falciparum malaria is one of the most widespread and deadliest infectious diseases in children under five years in endemic areas. The disease has been a strong force for evolutionary selection in the human genome, and uncovering the critical human genetic factors that confer resistance to the disease would provide clues to the molecular basis of protective immunity that would be invaluable for vaccine development. We investigated the effect of single nucleotide polymorphisms (SNPs) on malaria pathology in a case- control study of 1862 individuals from two major ethnic groups in three regions with intense perennial P. falciparum transmission in Cameroon. Twenty nine polymorphisms in cytokine and toll-like receptor (TLR) genes as well as the sickle cell trait (HbS) were assayed on the Sequenom iPLEX platform. Our results confirm the known protective effect of HbS against severe malaria and also reveal a protective effect of SNPs in interleukin-10 (IL10) cerebral malaria and hyperpyrexia. Furthermore, IL17RE rs708567 GA and hHbS rs334 AT individuals were associated with protection from uncomplicated malaria and anaemia respectively in this study. Meanwhile, individuals with the hHbS rs334 TT, IL10 rs3024500 AA, and IL17RD rs6780995 GA genotypes were more susceptible to severe malarial anaemia, cerebral malaria, and hyperpyrexia respectively. Taken together, our results suggest that polymorphisms in some immune response genes may have important implications for the susceptibility to severe malaria in Cameroonians. Moreover using uncomplicated malaria may allow us to identify novel pathways in the early development of the disease.     

Malaria eradication in Calabria, residual anopheles and transmission risk]

The paper is from a presentation at the meeting "La malaria in Calabria" (Cosenza, Italy, May 3, 1997) held to celebrate the half centenary of the antimalaria campaign (1947-1952) that eradicated the infection from the region. While Plasmodium falciparum had already disappeared in 1950, P. vivax persisted and the last autochtonous case was documented on the river Esaro in 1955. From the records of that time, it is clear that malaria eradication was achieved in the presence of important vector densities, as shown by larval surveys. Indoor DDT spraying, acting through the insecticide irritant effect, mainly avoided the house resting by the vector and forced the exposure of the mosquito to the outdoor environment where the lower mean temperatures eventually proved instrumental in destabilizing the parasite's life cycle. It is stressed that insecticide resistance never arised in the Anopheles vector populations of Calabria, in spite of at least ten years of continuous indoor spraying with an excellent overall coverage. This is expected in the absence of important selection pressures at the larval breeding sites and whenever the impact of the house spraying on the vector is due mainly to the insecticide irritant effect rather than to a direct imagocidal effect.     

Bioavailability and composition of dissolved organic carbon and nitrogen in a near coastal catchment of south-western Australia

The bioavailability and composition of dissolved organic carbon (DOC) and nitrogen (DON) were examined in 10 major sub-catchments of the Swan-Canning estuary, which bisects the city of Perth, in south-western Australia. Catchments contain a mix of forest, agriculture, and urban-dominated land-use, with the degree of development increasing near the city center. We incubated water samples from the 10 sub-catchments for 14 days at 25°C, and measured changes in DOC and DON and dissolved inorganic nitrogen (DIN). A greater proportion of DON (4–44%) was decomposed compared to DOC (1–17%). Both agricultural and urban catchments had high proportions of bioavailable DOC and DON, but overall DOC and DON losses were greatest in urban catchments. Using resin isolation techniques, we found that DOC was concentrated in the hydrophobic (humic) fraction, whereas DON had both hydrophobic and hydrophilic (non-humic) fractions. Hydrophobic DOC content was positively related to DOC decomposition. In contrast, DON decomposition was highly correlated with hydrophilic DON content and inversely related to the hydrophilic DOC/DON ratio, indicating a labile fraction of DON from non-humic sources. Taken together, these relationships suggest that bioavailable DOC may be supplied in part from terrestrial plant material, but bioavailable DON is likely to be from highly labile sources, possibly autochthonous or anthropogenic. Overall, labile DON was greater than initial DIN concentration at seven of ten sites and was even dominant in highly developed catchments. This study highlights the importance of organic N in urbanizing coastal catchments that, in addition to DIN, may serve as a readily available source of N for in-stream and estuarine production.     

Floodplain rehabilitation in North Cameroon: Impact on vegetation dynamics

Abstract. Since the construction in 1979 of a dam in the Logone floodplain in the Sahelo‐Sudanian zone of Cameroon, annual inundations have decreased, reducing perennial vegetation as important grazing source for nomadic herds and wildlife during the dry season. Presently, possibilities exist to release excess water for floodplain rehabilitation. In 1994 a pilot release was executed, reflooding 200 km², to verify predicted advantages. Vegetation has been studied from 1984 onwards along a transect covering flooded, recently reflooded and desiccated parts of the floodplain. Since 1993, the floristic composition has also been monitored in a grid in the centre of the impact zone. Cover of perennial grasses, most notably Echinochloa pyr amidalis and Oryza longistaminata increased from 41 to 61% in the reflooded zone. Vetiveria nigritana, a tussock grass that used to be dominant, disappeared slowly after the dam construction and has not shown a comeback. The cover of annual species, most notably Sorghum arundinaceum, a dominant annual grass only since the mid‐1980s, decreased in the reflooded zone from 58% to 34%. If the present conversion rate of annual into perennial grassland is extrapolated, recovery towards a 100% perennial state may be reached after the 2003 flooding season. Apart from favourable climatic conditions, recovery might be dependent on the restoration of soil fertility, limiting an approach focusing on flooding depth only.     

Density-dependence in parasite transmission dynamics

The transmission of vector-borne parasites is complex, yet to a large extent this complexity can be unravelled through the insights gained from simple mathematical models of the transmission system. The principle is simple because the key question is merely "what is the rate of increase in numbers of hosts affected?" Clearly, if this rate of increase is greater than unity then the infection can spread, while if it is less than unity it will decline. Ronald Ross in 1911 was the first to formulate this idea for malarial(1) and malaria transmission has since attracted most attention from modellers of parasitic diseases(2-4). But although it is implicitly recognized that nothing- not even parasitic transmission - can increase indefinitely, the importance of some degree of density-dependence in regulating the system tends to be neglected (see Box 1). In this article, Klaus Dietz explores some classical ideas of modelling parasitic disease transmission, emphasizing not only the importance of density dependence but also the importance of knowing exactly where such effects operate in the system.