Implementation of a novel PCR based method for detecting malaria parasites from naturally infected mosquitoes in Papua New Guinea

<p>Abstract</p> <p>Background</p> <p>Detection of <it>Plasmodium species </it>in mosquitoes is important for designing vector control studies. However, most of the PCR-based detection methods show some potential limitations. The objective of this study was to introduce an effective PCR-based method for detecting <it>Plasmodium vivax </it>and <it>Plasmodium falciparum </it>from the field-caught mosquitoes of Papua New Guinea.</p> <p>Methods</p> <p>A method has been developed to concurrently detect mitochondrial cytochrome b (<it>Cyt b</it>) of four human <it>Plasmodium </it>species using PCR (<it>Cytb</it>-PCR). To particularly discriminate <it>P. falciparum </it>from <it>P. vivax</it>, <it>Plasmodium ovale </it>and <it>Plasmodium malariae</it>, a polymerase chain reaction-repeated fragment length polymorphism (PCR-RFLP) has further been developed to use with this method. However, due to limited samples number of <it>P. ovale </it>and <it>P. malariae</it>; this study was mainly confined to <it>P. vivax </it>and <it>P. falciparum</it>. The efficiency of <it>Cytb</it>-PCR was evaluated by comparing it with two 'gold standards' enzyme linked immunosorbent assay specific for circumsporozoite protein (CS-ELISA) using artificially infected mosquitoes; and nested PCR specific for small subunit ribosomal RNA (<it>SSUrRNA</it>) using field caught mosquitoes collected from three areas (Kaboibus, Wingei, and Jawia) of the East Sepic Province of Papua New Guinea.</p> <p>Results</p> <p>A total of 90 mosquitoes were artificially infected with three strains of <it>Plasmodium</it>: <it>P. vivax-</it>210 (<it>n </it>= 30), <it>P. vivax</it>-247 (<it>n </it>= 30) and <it>P. falciparum </it>(<it>n </it>= 30). These infected mosquitoes along with another 32 unfed mosquitoes were first checked for the presence of <it>Plasmodium </it>infection by CS-ELISA, and later the same samples were compared with the <it>Cytb</it>-PCR. CS-ELISA for <it>P. vivax</it>-210, <it>P. vivax</it>-247 and <it>P. falciparum </it>detected positive infection in 30, 19 and 18 mosquitoes respectively; whereas <it>Cytb</it>-PCR detected 27, 16 and 16 infections, respectively. The comparison revealed a close agreement between the two assays (κ = 0.862, 0.842 and 0.894, respectively for Pv-210, Pv-247 and <it>P. falciparum </it>groups). It was found that the eight CS-ELISA-positive mosquitoes detected negative by <it>Cytb</it>-PCR were false-positive results. The lowest detection limit of this <it>Cytb</it>-PCR was 10 sporozoites. A highly concordance result was also found between nested PCR and <it>Cytb</it>-PCR using 107 field caught mosquitoes, and both tests concordantly detected <it>P. falciparum </it>in an <it>Anopheles punctulatus </it>mosquito collected from Kaboibus. Both tests thus suggested an overall sporozoite rate of 0.9% (1/107) in the study areas. Subsequently, PCR-RFLP efficiently discriminated <it>P. falciparum </it>from <it>P. vivax </it>for all of the <it>Cytb</it>-PCR positive samples.</p> <p>Conclusion</p> <p>A single step PCR based method has been introduced here that is highly sensitive, efficient and reliable for identifying <it>P. vivax </it>and <it>P. falciparum </it>from mosquitoes. The reliability of the technique was confirmed by its ability to detect <it>Plasmodium </it>as efficiently as those of CS-ELISA and nested PCR. Application of the assay offers the opportunity to detect vector species of Papua New Guinea and may contribute for designing further vector control programmes.</p

Changes in body composition parameters with age

Health risks are associated with changes in body composition parameters with age. In the present study, body composition parameters （appendicular skeletal muscle mass ［ASMM］, fat mass ［FM］, and water content ［water］） using bioelectrical impedance analysis （BIA） and total skeletal MM （TSMM） measured by 24-h creatinine excretion （Cr） were obtained in 30 male and 38 female healthy subjects. BIA-ASMM in both sexes and Cr-TSMM in females were negatively correlated with aging, and BIA-FM was negatively correlated with BIA-water in both sexes. Of note, Cr-TSMM was a more sensitive marker of MM than BIA-ASMM. Thus, decreases in BIA-ASMM and Cr- TSMM were the most consistent markers of aging and sarcopenia. This study may help promote nursing care for healthy aging

Modelling the dynamics and control of Schistosoma japonicum transmission on Bohol island, the Philippines

We have investigated a mathematical model for the transmission of Schistosoma japonicum in the infested region of northeastern Bohol island in the Philippines. The development of transmission models is important for planning control strategies. Since S. japonicum has a complicated mode of transmission, the rates of transmission among its hosts cannot be measured directly by field observation. Instead, they have been estimated through model analysis. The model takes into account the seasonal variations and includes a function of control measures. In 1981, a project to eliminate schistosomiasis started on Bohol island. The prevalence decreased dramatically and has kept low level less than 1%. The simulations based on the model predicted that there is little probability of resurgence of an epidemic in the northeastem endemic villages of Bohol island due to the fact that the project has attained a high coverage of selective mass treatment based oil stool examination accompanied by a successful snail control operation. (c) 2005 Elsevier Ireland Ltd. All rights reserved.</p

Superhydrophilicity on nano-rough carbon surfaces achieved by hyperthermal oxygen-atom beam exposure

In order to investigate a method to increase hydrophilicity on nano-rough carbon surfaces, a nano-rough surface of C(60) film and an atomically flat surface of highly oriented pyrolytic graphite (HOPG) were oxidized by hyperthermal oxygen-atom beam exposure and the hydrophilicities of the surfaces were investigated. Superhydrophilicity were achieved on these exposed carbon surfaces, which had low O/C ratio of approximately 28% and surface roughness (Ra) of approximately 3 nm. The direct oxidations on sp(2) bonded carbon atoms (basal plane) of these two carbon materials by the exposure of hyperthermal O-atom beam would contribute the superhydrophilicity

Modeling the dynamics and control of Schistosoma japonicum transmission on Bohol Island, the Philippines. Parasitol. Int

Abstract We have investigated a mathematical model for the transmission of Schistosoma japonicum in the infested region of northeastern Bohol island in the Philippines. The development of transmission models is important for planning control strategies. Since S. japonicum has a complicated mode of transmission, the rates of transmission among its hosts cannot be measured directly by field observation. Instead, they have been estimated through model analysis. The model takes into account the seasonal variations and includes a function of control measures. In 1981, a project to eliminate schistosomiasis started on Bohol island. The prevalence decreased dramatically and has kept low level less than 1%. The simulations based on the model predicted that there is little probability of resurgence of an epidemic in the northeastern endemic villages of Bohol island due to the fact that the project has attained a high coverage of selective mass treatment based on stool examination accompanied by a successful snail control operation