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1.
Parasit Vectors ; 12(1): 399, 2019 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-31409374

RESUMO

BACKGROUND: Sampling methodologies for mosquitoes that are capable of transmitting vector-borne infectious diseases provide critical information on entomological endpoints. Reliable and meaningful field data is vital to the understanding of basic vector biology as well as disease transmission. Various traps take advantage of different vector behaviors and are inevitably subject to sampling biases. This study represents the first comparison of kelambu traps (KT) to barrier screens (BS), barrier screens with eaves (BSE) and indoor and outdoor human landing catches (HLCs). METHODS: Two trap comparison studies were undertaken. In the first study, mosquitoes were collected in Karama over 26 trapping nights to evaluate the kelambu trap relative to indoor and outdoor HLCs. In the second study, mosquitoes were collected in Karama over 12 trapping nights to compare the kelambu trap, barrier screen, barrier screen with eaves and outdoor HLCs. The kelambu trap, barrier screen and barrier screen with eaves obstruct the flight of mosquitos. HLCs target host-seeking behaviors. RESULTS: There was no significant difference between indoor and outdoor HLCs for overall Anopheles mosquito abundance. All five of the molecularly identified Anopheles species collected by HLCs, An. aconitus, An. barbirostris, An. peditaeniatus, An. vagus and An. tessellatus, are reported as vectors of malaria in Indonesia. The kelambu trap (n = 2736) collected significantly more Anopheles mosquitoes than indoor HLCs (n = 1286; Z = 3.193, P = 0.004), but not the outdoor HLCs (n = 1580; Z = 2.325, P = 0.053). All traps collected statistically similar abundances for the primary species, An. barbirostris. However, both comparison studies found significantly higher abundances for the kelambu trap for several secondary species compared to all other traps: An. nigerriumus, An. parangensis, An. tessellatus and An. vagus. The kelambu trap retained the highest species richness and Gini-Simpson's diversity index for both comparison studies. CONCLUSIONS: This study demonstrates that the kelambu trap collects overall Anopheles abundance and species-specific abundances at statistically similar or higher rates than HLCs in Sulawesi, Indonesia. Therefore, the kelambu trap should be considered as an exposure-free alternative to HLCs for research questions regarding Anopheles species in this malaria endemic region.


Assuntos
Anopheles , Comportamento Alimentar , Controle de Mosquitos/métodos , Mosquitos Vetores , Animais , Entomologia/instrumentação , Entomologia/métodos , Indonésia , Especificidade da Espécie
2.
Parasit Vectors ; 11(1): 440, 2018 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-30064507

RESUMO

BACKGROUND: Mosquito sampling methods target different aspects of mosquito behavior and are subject to trap and location specific biases. The barrier screen sampling method was developed and tested to sample free-flying, blood-fed, and host-seeking mosquitoes. During a pilot study, this method was useful in obtaining an unbiased sample of mosquitoes flying between outdoor larval habitats, and sites where blood meals were obtained. However, a relatively small number of blood-fed Anopheles mosquitoes were collected in Indonesia during the pilot study. The sampling method was extended in South Lampung, Indonesia, to enable the collection of blood-fed mosquitoes. This study aimed to intercept mosquitoes flying between human habitations and larval habitats with a barrier screen and to characterize mosquito composition, flight characteristics (direction, height and time), abdominal status, and parity. RESULTS: Barrier screens intercepted 15 different mosquito species in South Lampung: eight Anopheles spp. and seven Culex spp. Species compositions varied among the villages in South Lampung. About 15% of Anopheles spp. caught were blood-fed, of which 28.2% of those tested had fed on humans. This is the first time human blood-fed anophelines have been collected in Indonesia using barrier screens. Blood meals identified included cow, dog, goat, and human, as well as mixed blood meals. Activity of unfed An. subpictus, the primary vector collected, flying towards human habitations peaked between 20:00-12:00 h, with a slow decline in activity until 18:00 h. Unfed and fed An. sundaicus, had a different activity profile compared to An. subpictus. Other species demonstrated varied peak activity times, with earlier activity occurring as a general trend. For the Anopheles mosquitoes collected, 55.5% were collected below 0.5 m and 83.9% were captured resting < 1 m from the ground. Parity dissections enabled age structure by species, which revealed species-specific traits such as nulliparous An. subpictus being more active early in the night relative to An. sundaicus. CONCLUSIONS: This study demonstrates that barrier screens are an effective mosquito sampling method that can be used to gain insights into local mosquito species composition, flight characteristics (direction, height and time), abdominal status, and parity.


Assuntos
Anopheles/fisiologia , Comportamento Animal/fisiologia , Culex/fisiologia , Abdome , Animais , Sangue , Feminino , Indonésia , Projetos Piloto , Especificidade da Espécie , Fatores de Tempo
3.
Nat Prod Res ; 32(17): 2067-2070, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28768428

RESUMO

The rapid emergence of antimalarial drug resistance necessitates a continual effort on novel drug discovery. A cyanobacterium, Spirulina platensis, is a potential antimalarial agent that has been widely consumed as food supplement in the form of crude extract. It is known to possess antiviral, antibacterial and antifungi activities. This study examined the antimalarial activities of several Spirulina formulas against Plasmodium falciparum 3D7, in vitro. The tested Spirulina formulas included commercially available capsule, crude extract and alkaloid fraction. Results showed that all tested formula possessed antimalarial activities with the Spirulina capsule exhibited the highest activities (IC50 = 2.16 µg/mL). Light and electron microscopies revealed interference of the Spirulina with the parasite hemozoin formation. In conclusion, all tested Spirulina formulas and fraction exhibited moderate to high antimalarial activities.


Assuntos
Antimaláricos/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Spirulina/química , Composição de Medicamentos , Hemeproteínas/efeitos dos fármacos , Microscopia
4.
Adv Biomed Res ; 5: 104, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27376043

RESUMO

BACKGROUND: Cell culture techniques have many advantages for investigation of drug transport to target organ like liver. HepG2 and Huh-7 are two cell lines available from hepatoma that can be used as a model for hepatic drug transport. The present study is aimed to analyze the expression level of several drug transporter genes in two hepatoma cell lines, HepG2 and Huh-7 and their response to inhibitors. MATERIALS AND METHODS: This is an in vitro study using HepG2 and Huh-7 cells. The expression level of the following drug transporter genes was quantified: P-glycoprotein/multidrug resistance protein 1, Organic Anionic Transporter Protein 1B1 (OATP1B1) and Organic Cationic Transporter-1 (OCT1). Ribonucleic acid was extracted from the cells using Tripure isolation reagent, then gene expression level of the transporters is quantified using Applied Biosystems quantitative reverse transcriptase polymerase chain reaction. Verapamil (P-glycoprotein inhibitor), nelfinavir (OATP1B1 inhibitor), quinidine (OCT1 inhibitor) were used to differentiate the inhibitory properties of these agents to the transporter expressions in HepG2 and Huh-7 cells. RESULTS: Huh-7 shows a higher level of P-glycoprotein, OATP1B1 and OCT1 expressions compared with those of HepG2. Verapamil reduces the expressions of P-glycoprotein in HepG2 and Huh-7; nelfinavir reduces the expression of OATP1B1 in HepG2 and Huh-7; while quinidine reduces the OCT1 gene expressions in HepG2, but not in Huh-7 cells. CONCLUSION: This study indicates that HepG2 might be a more suitable in vitro model than Huh-7 to study drug transport in hepatocytes involving drug transporters.

5.
Malar J ; 15: 192, 2016 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-27060058

RESUMO

BACKGROUND: Members of the Anopheles punctulatus group dominate Papua, Indonesia and Papua New Guinea (PNG), with a geographic range that extends south through Vanuatu. An. farauti and An. punctulatus are the presumed major vectors in this region. Although this group of species has been extensively studied in PNG and the southern archipelagoes within their range, their distribution, ecology and vector behaviours have not been well characterized in eastern Indonesia. METHODS: Mosquitoes were collected in five villages in Jayapura province, Papua, Indonesia using human-landing collections, animal-baited tents and backpack aspirators. Mosquitoes were morphologically typed and then molecularly distinguished based on ribosomal ITS2 sequences and tested for Plasmodium falciparum and P. vivax infection using circumsporozoite ELISA and PCR. RESULTS: The presence and vector status of An. farauti 4 in Papua, Indonesia is confirmed here for the first time. The data indicate that this species is entering houses at a rate that increases its potential to come into contact with humans and act as a major malaria vector. An. farauti 4 was also abundant outdoors and biting humans during early evening hours. Other species collected in this area include An. farauti 1, An. hinesorum, An. koliensis, An. punctulatus, and An. tessellatus. Proboscis morphology was highly variable within each species, lending support to the notion that this characteristic is not a reliable indicator to distinguish species within the An. punctulatus group. CONCLUSIONS: The vector composition in Papua, Indonesia is consistent with certain northern areas of PNG, but the behaviours of anophelines sampled in this region, such as early and indoor human biting of An. farauti 4, may enable them to act as major vectors of malaria. Presumed major vectors An. farauti and An. punctulatus were not abundant among these samples. Morphological identification of anophelines in this sample was often inaccurate, highlighting the importance of using molecular analysis in conjunction with morphological investigations to update keys and training tools.


Assuntos
Anopheles/classificação , Anopheles/fisiologia , Comportamento Alimentar , Insetos Vetores , Plasmodium falciparum/isolamento & purificação , Plasmodium vivax/isolamento & purificação , Animais , Anopheles/anatomia & histologia , Anopheles/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Indonésia , Análise de Sequência de DNA
6.
Malar J ; 15: 218, 2016 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-27083152

RESUMO

BACKGROUND: The Indonesian archipelago is endemic for malaria. Although Plasmodium falciparum and P. vivax are the most common causes for malaria cases, P. malariae and P. ovale are also present in certain regions. Zoonotic case of malaria had just became the attention of public health communities after the Serawak study in 2004. However, zoonotic case in Indonesia is still under reported; only one published report of knowlesi malaria in South Kalimantan in 2010. CASE PRESENTATION: A case of Plasmodium knowlesi infection in a worker from a charcoal mining company in Central Kalimantan, Indonesia was described. The worker suffered from fever following his visit to a lowland forest being cut and converted into a new mining location. CONCLUSION: This study confirmed a zoonotic infection using polymerase chain reaction amplification and Sanger sequencing of plasmodial DNA encoding the mitochondrial cytochrome c oxidase subunit I (mtCOI).


Assuntos
Malária/diagnóstico , Plasmodium knowlesi/isolamento & purificação , Zoonoses/diagnóstico , Animais , Bornéu , Complexo IV da Cadeia de Transporte de Elétrons/análise , Complexo IV da Cadeia de Transporte de Elétrons/genética , Humanos , Indonésia , Malária/parasitologia , Masculino , Pessoa de Meia-Idade , Proteínas Mitocondriais/análise , Proteínas Mitocondriais/genética , Filogenia , Plasmodium knowlesi/genética , Reação em Cadeia da Polimerase , Proteínas de Protozoários/análise , Proteínas de Protozoários/genética , Análise de Sequência de DNA , Zoonoses/parasitologia
7.
PLoS One ; 11(3): e0150577, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26939002

RESUMO

The emergence of insecticide resistant Aedes aegypti mosquitoes has hampered dengue control efforts. WHO susceptibility tests, using several pyrethroid compounds, were conducted on Ae. aegypti larvae that were collected and raised to adulthood from Semarang, Surakarta, Kudus and Jepara in Java. The AaNaV gene fragment encompassing kdr polymorphic sites from both susceptible and resistant mosquitoes was amplified, and polymorphisms were associated with the resistant phenotype. The insecticide susceptibility tests demonstrated Ae, aegypti resistance to the pyrethroids, with mortality rates ranging from 1.6%-15.2%. Three non-synonymous polymorphisms (S989P, V1016G and F1534C) and one synonymous polymorphism (codon 982) were detected in the AaNaV gene. Eight AaNaV alleles were observed in specimens from Central Java. Allele 3 (SGF) and allele 7 (PGF) represent the most common alleles found and demonstrated strong associations with resistance to pyrethroids (OR = 2.75, CI: 0.97-7.8 and OR = 7.37, CI: 2.4-22.5, respectively). This is the first report of 8 Ae. aegypti AaNaV alleles, and it indicates the development of resistance in Ae. aegypti in response to pyrethroid insecticide-based selective pressure. These findings strongly suggest the need for an appropriate integrated use of insecticides in the region. The 989P, 1016G and 1534C polymorphisms in the AaNaV gene are potentially valuable molecular markers for pyrethroid insecticide resistance monitoring.


Assuntos
Aedes/genética , Dengue/genética , Genética Populacional , Resistência a Inseticidas/genética , Canais de Sódio Disparados por Voltagem/genética , Aedes/efeitos dos fármacos , Aedes/virologia , Alelos , Animais , Dengue/transmissão , Dengue/virologia , Humanos , Indonésia , Inseticidas/farmacologia , Piretrinas/farmacologia
8.
Am J Trop Med Hyg ; 81(2): 366-9, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19635900

RESUMO

Microscopic detection of Plasmodium vivax gametocytes, the sexual life stage of this malaria parasite, is insensitive because P. vivax parasitaemia is low. To detect and quantify gametocytes a more sensitive, quantitative real-time Pvs25-QT-NASBA based on Pvs25 mRNA was developed and tested in two clinical sample sets from three different continents. Pvs25-QT-NASBA is highly reproducible with low inter-assay variation and reaches sensitivity approximately 800 times higher than conventional microscopic gametocyte detection. Specificity was tested in 104 samples from P. vivax-, P. falciparum-, P. malariae-, and P. ovale-infected patients. All non-vivax samples were negative in the Pvs25-QT-NASBA; out of 74 PvS18-QT-NASBA positive samples 69% were positive in the Pvs25-QT-NASBA. In a second set of 136 P. vivax microscopically confirmed samples, gametocyte prevalence was 8%, whereas in contrast 66% were positive by Pvs25-QT-NASBA. The data suggest that the human P. vivax gametocyte reservoir is much larger when assessed by Pvs25-QT-NASBA than by microscopy.


Assuntos
Malária Vivax/parasitologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Plasmodium vivax/isolamento & purificação , Animais , Sequência de Bases , Humanos , Plasmodium vivax/citologia , Plasmodium vivax/genética , Proteínas de Protozoários/genética , RNA/genética , RNA de Protozoário/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
9.
Biol Pharm Bull ; 29(5): 1050-2, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16651745

RESUMO

Malaria is one of the most life-threatening infectious diseases worldwide and claims millions of people's lives each year. The appearance of drug-resistance Plasmodium falciparum has made the treatment of malaria increasingly problematic, and thus, it is a dire need to search the new alternatives of current drugs. In the present study, 44 cassane- and norcassane-type diterpenes isolated from Caesalpinia crista of Myanmar and Indonesia were evaluated for their antimalarial activity against the malaria parasite Plasmodium falciparum FCR-3/A2 clone in vitro. Most of the tested diterpenes displayed antimalarial activity, and norcaesalpinin E (28) showed the most potent activity with an IC50 value of 0.090 microM, more potent than the clinically used drug chloroquine (IC50, 0.29 microM). Based on the observed results, a structure-activity relationship has been established.


Assuntos
Antimaláricos/farmacologia , Caesalpinia/química , Diterpenos/farmacologia , Animais , Cloroquina/farmacologia , Diterpenos/química , Eritrócitos/parasitologia , Humanos , Técnicas In Vitro , Indonésia , Mianmar , Plasmodium falciparum/efeitos dos fármacos , Relação Estrutura-Atividade
10.
J Nat Prod ; 68(5): 706-10, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15921414

RESUMO

The CH2Cl2 extract of the seed kernels of Caesalpinia crista, which exhibited promising antimalarial activity against Plasmodium berghei-infected mice in vivo, was examined and resulted in the isolation of seven new furanocassane-type diterpenes [caesalpinins C-G (1-5) and norcaesalpinins D and E (6, 7)] together with norcaesalpinins A-C (8-10) and 11 known compounds (norcaesalpinins A-C, 2-acetoxy-3-deacetoxycaesaldekarin e, caesalmin B, caesaldekarin e, caesalpin F, 14(17)-dehydrocaesalpin F, 2-acetoxycaesaldekarin e, 7-acetoxybonducellpin C, and caesalmin G). Their structures were determined on the basis of spectroscopic analysis. The isolated diterpenes showed significant dose-dependent inhibitory effects on Plasmodium falciparum FCR-3/A2 growth in vitro. Their IC50 values ranged from 90 nM to 6.5 microM, and norcaesalpinin E (7) showed the most potent inhibitory activity (IC50, 90 nM).


Assuntos
Antimaláricos/isolamento & purificação , Caesalpinia/química , Diterpenos/isolamento & purificação , Plantas Medicinais/química , Animais , Antimaláricos/química , Antimaláricos/farmacologia , Diterpenos/química , Diterpenos/farmacologia , Indonésia , Concentração Inibidora 50 , Camundongos , Estrutura Molecular , Plasmodium berghei/efeitos dos fármacos , Plasmodium falciparum/efeitos dos fármacos , Sementes/química
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