RESUMO
In an area endemic with Indian visceral leishmaniasis (VL), we performed direct xenodiagnosis to evaluate the transmission of Leishmania donovani from patients with VL-human immunodeficiency virus (HIV) coinfection to the vector sandflies, Phlebotomus argentipes. Fourteen patients with confirmed VL-HIV coinfection, with a median parasitemia of 42 205 parasite genome/mL of blood, were exposed to 732 laboratory-reared pathogen-free female P argentipes sandflies on their lower arms and legs. Microscopy revealed that 16.66% (122/732) of blood-fed flies were xenodiagnosis positive. Notably, 93% (13/14) of the VL-HIV group infected the flies, as confirmed by quantitative polymerase chain reaction and/or microscopy, and were 3 times more infectious than those who had VL without HIV.
Assuntos
Coinfecção , Infecções por HIV , Leishmania donovani , Leishmaniose Visceral , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/complicações , Animais , Humanos , Índia/epidemiologia , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Feminino , Adulto , Coinfecção/virologia , Coinfecção/epidemiologia , Coinfecção/parasitologia , Leishmania donovani/isolamento & purificação , Masculino , Phlebotomus/parasitologia , Phlebotomus/virologia , Doenças Endêmicas , Pessoa de Meia-Idade , Adulto Jovem , Xenodiagnóstico , Insetos Vetores/parasitologia , Insetos Vetores/virologia , AdolescenteRESUMO
Begomoviruses are the largest group of plant viruses transmitted exclusively by the whitefly, Bemisia tabaci (Gennadius), in a persistent, circulative, and nonpropagative manner. Begomoviruses in association with B. tabaci cause enormous loss to world agricultural crops. Transmission, retention, and circulation of begomovirus in B. tabaci are facilitated by its interaction with several proteins of the insect and its endosymbionts. However, very few such proteins have been identified from B. tabaci that are involved in this specific interaction. Here, we have performed yeast two-hybrid assay between B. tabaci complementary DNA expression library and the coat protein (CP) of tomato leaf curl New Delhi virus (ToLCNDV) and cotton leaf curl Rajasthan virus (CLCuV). Collagen was the common protein found to be interacting with both of the viruses. The collagen protein was found to be localized in gut layers of B. tabaci. Additionally, pull-down and dot-blot assays confirmed the association of endogenous collagen with ToLCNDV CP. Immunolocalization analysis also showed colocalization of ToLCNDV particles and collagen within insect gut. Finally, B. tabaci fed on anticollagen antibody and exhibited â¼46% reduction in ToLCNDV transmission, suggesting a supportive role for collagen in virus transmission.
Assuntos
Begomovirus , Hemípteros , Doenças das Plantas/virologia , Animais , Begomovirus/patogenicidade , Colágeno , Hemípteros/virologia , ÍndiaRESUMO
Begomoviruses are a major group of plant viruses, transmitted exclusively by Bemisia tabaci (Gennadius) in a persistent circulative non-propagative manner. The information regarding molecular and cellular basis underlying Begomovirus - whitefly interaction is very scarce. Evidences have suggested that the insect gut possesses some crucial protein receptors that allow specific entry of virus into the insect haemolymph. We have performed yeast two hybrid gut cDNA expression library screening against coat protein of Tomato leaf curl New Delhi virus (ToLCV) and Cotton leaf curl Rajasthan virus (CLCuV) as bait. Midgut protein (MGP) was the common protein found interacting with both ToLCV and CLCuV. MGP was localized in whole mount B. tabaci as well as in dissected guts through confocal microscopy. Pull down and dot blot assays confirmed in vitro interaction between ToLCV/CLCuV coat protein and MGP. Immunolocalization analysis also showed colocalization of ToLCV/CLCuV particles and MGP within insect's gut. Finally, anti-MGP antibody fed B. tabaci, exhibited 70% reduction in ToLCV transmission, suggesting a supportive role for MGP in virus transmission.
Assuntos
Begomovirus/metabolismo , Proteínas de Insetos/biossíntese , Doenças das Plantas/virologia , Simbiose/genética , Animais , Begomovirus/patogenicidade , Sistema Digestório/virologia , Hemípteros/metabolismo , Hemípteros/virologia , Hemolinfa/metabolismo , Hemolinfa/virologia , Índia , Proteínas de Insetos/metabolismo , Insetos Vetores/metabolismo , Insetos Vetores/virologia , Doenças das Plantas/genética , Internalização do VírusRESUMO
BACKGROUND: In Southeast Asia, treatment is recommended for all patients with post-kala-azar dermal leishmaniasis (PKDL). Adherence to the first-line regimen, twelve weeks of miltefosine (MF), is low and ocular toxicity has been observed with this exposure period. We assessed the safety and efficacy of two shorter-course treatments: liposomal amphotericin B (LAmB) alone and combined with MF. METHODOLOGY/PRINCIPAL FINDINGS: An open-label, phase II, randomized, parallel-arm, non-comparative trial was conducted in patients with parasitologically confirmed PKDL, 6 to ≤60 years. Patients were assigned to 20 mg/kg LAmB (total dose, in five injections over 15 days) alone or combined with allometric MF (3 weeks). The primary endpoint was definitive cure at 12 months, defined as complete resolution of papular and nodular lesions and >80% re-pigmentation of macular lesions. Definitive cure at 24 months was a secondary efficacy endpoint. 118/126 patients completed the trial. Definitive cure at 12 months was observed in 29% (18/63) patients receiving LAmB and 30% (19/63) receiving LAmB/MF (mITT), increasing to 58% and 66%, respectively, at 24 months. Most lesions had resolved/improved at 12 and 24 months for patients receiving LAmB (90%, 83%) and LAmB/MF (85%, 88%) by qualitative assessment. One death, unrelated to study drugs, was reported; no study drug-related serious adverse events were observed. The most frequent adverse drug reactions were MF-related vomiting and nausea, and LAmB-related hypokalaemia and infusion reactions. Most adverse events were mild; no ocular adverse events occurred. CONCLUSIONS/SIGNIFICANCE: Both regimens are suitably safe and efficacious alternatives to long-course MF for PKDL in South Asia. TRIAL REGISTRATION: CTRI/2017/04/008421.
Assuntos
Anfotericina B , Antiprotozoários , Leishmaniose Cutânea , Leishmaniose Visceral , Fosforilcolina , Humanos , Anfotericina B/uso terapêutico , Anfotericina B/efeitos adversos , Anfotericina B/administração & dosagem , Fosforilcolina/análogos & derivados , Fosforilcolina/uso terapêutico , Fosforilcolina/administração & dosagem , Fosforilcolina/efeitos adversos , Bangladesh , Masculino , Antiprotozoários/uso terapêutico , Antiprotozoários/efeitos adversos , Antiprotozoários/administração & dosagem , Adulto , Adolescente , Feminino , Pessoa de Meia-Idade , Adulto Jovem , Criança , Índia , Leishmaniose Visceral/tratamento farmacológico , Resultado do Tratamento , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/parasitologia , Quimioterapia CombinadaRESUMO
Visceral leishmaniasis (VL) is a vector-borne protozoan disease, which can be fatal if left untreated. Synthetic chemical insecticides are very effective tools for controlling of insect vectors, including the sand fly Phlebotomus argentipes, the vector of VL in the Indian subcontinent. However, repeated use of the same insecticide with increasing doses potentially can create high selection pressure and lead to tolerance and resistance development. The objective of this study was to determine the lethal concentrations and assess levels of susceptibility, diagnostic doses and times to death of laboratory-reared P. argentipes to five insecticides that are used worldwide to control vectors. Using the Center for Disease Control and Prevention (CDC) bottle bioassay, 20-30 sand flies were exposed in insecticide- coated 500-ml glass bottles. Flies were then observed for 24 hours and mortality was recorded. Dose-response survival curves were generated for each insecticide using QCal software and lethal concentrations causing 50%, 90% and 95% mortality were determined. A bioassay was also conducted to determine diagnostic doses and diagnostic times by exposing 20-30 flies in each bottle containing set concentrations of insecticide. Mortality was recorded at 10-minute intervals for 120 minutes to generate the survival curve. Phlebotomus argentipes are highly susceptible to alpha-cypermethrin, followed by deltamethrin, malathion, chlorpyrifos, and least susceptible to DDT. Also, the lowest diagnostic doses and diagnostic times were established for alpha-cypermethrin (3µg/ml for 40 minutes) to kill 100% of the flies. The susceptibility data, diagnostic doses and diagnostic times presented here will be useful as baseline reference points for future studies to assess insecticide susceptibility and resistance monitoring of field caught sand flies and to assist in surveillance as VL elimination is achieved in the region.
Assuntos
Inseticidas , Leishmaniose Visceral , Phlebotomus , Psychodidae , Animais , Estados Unidos , Inseticidas/farmacologia , Phlebotomus/fisiologia , Leishmaniose Visceral/prevenção & controle , Resistência a Inseticidas , Índia , Bioensaio , Centers for Disease Control and Prevention, U.S.RESUMO
[This corrects the article DOI: 10.1371/journal.pntd.0010613.].
RESUMO
Leishmaniasis on the Indian subcontinent is thought to have an anthroponotic transmission cycle. There is no direct evidence that a mammalian host other than humans can be infected with Leishmania donovani and transmit infection to the sand fly vector. The aim of the present study was to evaluate the impact of sand fly feeding on other domestic species and provide clinical evidence regarding possible non-human reservoirs through experimental sand fly feeding on cows, water buffalo goats and rodents. We performed xenodiagnosis using colonized Phlebotomus argentipes sand flies to feed on animals residing in villages with active Leishmania transmission based on current human cases. Xenodiagnoses on mammals within the endemic area were performed and blood-fed flies were analyzed for the presence of Leishmania via qPCR 48hrs after feeding. Blood samples were also collected from these mammals for qPCR and serology. Although we found evidence of Leishmania infection within some domestic mammals, they were not infectious to vector sand flies. Monitoring infection in sand flies and non-human blood meal sources in endemic villages leads to scientific proof of exposure and parasitemia in resident mammals. Lack of infectiousness of these domestic mammals to vector sand flies indicates that they likely play no role, or a very limited role in Leishmania donovani transmission to people in Bihar. Therefore, a surveillance system in the peri-/post-elimination phase of visceral leishmaniasis (VL) must monitor absence of transmission. Continued surveillance of domestic mammals in outbreak villages is necessary to ensure that a non-human reservoir is not established, including domestic mammals not present in this study, specifically dogs.
Assuntos
Leishmania donovani , Leishmaniose Visceral , Leishmaniose , Phlebotomus , Psychodidae , Feminino , Bovinos , Humanos , Cães , Animais , Leishmaniose Visceral/epidemiologia , Gado , RoedoresRESUMO
BACKGROUND: Visceral leishmaniasis, also known on the Indian subcontinent as kala-azar, is a fatal form of leishmaniasis caused by the protozoan parasite Leishmania donovani and transmitted by the bites of the vector sandfly Phlebotomus argentipes. To achieve and sustain elimination of visceral leishmaniasis, the transmission potential of individuals exposed to L donovani from across the infection spectrum needs to be elucidated. The aim of this study was to evaluate the relative infectiousness to the sandfly vector of patients with visceral leishmaniasis or post-kala-azar dermal leishmaniasis, before and after treatment, and individuals with asymptomatic infection. METHODS: In this prospective xenodiagnosis study done in Muzaffarpur district of Bihar, India, we included patients with clinically confirmed active visceral leishmaniasis or post-kala-azar dermal leishmaniasis who presented to the Kala-Azar Medical Research Center. These participants received treatment for L donovani infection. We also included asymptomatic individuals identified through a serosurvey of 17â254 people living in 26 high-transmission clusters. Eligible participants were aged 12-64 years, were HIV negative, and had clinically or serologically confirmed L donovani infection. During xenodiagnosis, the forearms or lower legs of participants were exposed to 30-35 female P argentipes sandflies for 30 min. Blood-engorged flies were held in an environmental cabinet at 28°C and 85% humidity for 60-72 h, after which flies were dissected and evaluated for L donovani infection by microscopy and quantitative PCR (qPCR). The primary endpoint was the proportion of participants with visceral leishmaniasis or post-kala-azar dermal leishmaniasis, before and after treatment, as well as asymptomatic individuals, who were infectious to sandflies, with a participant considered infectious if promastigotes were observed in one or more individual flies by microscopy, or if one or more of the pools of flies tested positive by qPCR. FINDINGS: Between July 12, 2016, and March 19, 2019, we recruited 287 individuals, including 77 with active visceral leishmaniasis, 26 with post-kala-azar dermal leishmaniasis, and 184 with asymptomatic infection. Of the patients with active visceral leishmaniasis, 42 (55%) were deemed infectious to sandflies by microscopy and 60 (78%) by qPCR before treatment. No patient with visceral leishmaniasis was found to be infectious by microscopy at 30 days after treatment, although six (8%) were still positive by qPCR. Before treatment, 11 (42%) of 26 patients with post-kala-azar dermal leishmaniasis were deemed infectious to sandflies by microscopy and 23 (88%) by qPCR. Of 23 patients who were available for xenodiagnosis after treatment, one remained infectious to flies by qPCR on the pooled flies, but none remained positive by microscopy. None of the 184 asymptomatic participants were infectious to sandflies. INTERPRETATION: These findings confirm that patients with active visceral leishmaniasis and patients with post-kala-azar dermal leishmaniasis can transmit L donovani to the sandfly vector and suggest that early diagnosis and treatment could effectively remove these individuals as infection reservoirs. An important role for asymptomatic individuals in the maintenance of the transmission cycle is not supported by these data. FUNDING: Bill & Melinda Gates Foundation.
Assuntos
Leishmaniose Visceral , Phlebotomus , Psychodidae , Animais , Infecções Assintomáticas , Feminino , Humanos , Índia/epidemiologia , Leishmaniose Visceral/diagnóstico , Masculino , Phlebotomus/parasitologia , Estudos Prospectivos , Psychodidae/parasitologia , XenodiagnósticoRESUMO
BACKGROUND: Visceral leishmaniasis (VL) is a deadly vector-borne disease. Approximately 90% of Indian VL cases occur in Bihar, where the sand fly, Phlebotomus argentipes, is the principal vector. Sand fly control in Bihar consists of indoor residual spraying (IRS), the practice of spraying the inner walls of village dwellings with insecticides. Prior researchers have evaluated success of IRS-control by estimating vector abundance in village houses, but the number of sampling periods (n = 2-3) were minimal, and outdoor-resting P. argentipes were neglected. We describe a large-scale field study, performed in 24 villages within two Bihari districts, during which P. argentipes were collected biweekly over 47-weeks, in cattle enclosures, houses, and outdoors in peri-domestic vegetation. The objectives of this study were to provide updated P. argentipes ecological field data, and determine if program-initiated IRS-treatment had led to noticeable differences in vector abundance. PRINCIPAL FINDINGS: P. argentipes (n = 126,901) relative abundance was greatest during the summer months (June-August) when minimum temperatures were highest. P. argentipes were most frequently collected from cattle enclosures (~46% total; ~56% blood fed). Many sand flies were found to have taken blood from multiple sources, with ~81% having blood fed on humans and ~60% blood feeding on bovines. Nonparametric statistical tests were determined most appropriate for evaluating IRS-treatment. Differences in P. argentipes abundance in houses, cattle enclosures and vegetation were detected between IRS-treated and untreated villages in only ~9% of evaluation periods occurring during the peak period of human-vector exposure (June-August) and in ~8% of the total observations. No significant differences were detected between the numbers of P. argentipes collected in vegetation close to the experimental villages. CONCLUSION: The results of this study provide updated data regarding P. argentipes seasonal abundance, spatial distribution, and host preferances, and suggest vector abundance has not significantly declined in IRS-treated villages. We suggest that IRS be supplemented with vector control strategies targeting exophagic, exophilic P. argentipes, and that disease surveillance be accompanied by rigorous vector population monitoring.
Assuntos
Controle de Insetos/métodos , Insetos Vetores/efeitos dos fármacos , Inseticidas/farmacologia , Leishmaniose Visceral/prevenção & controle , Leishmaniose Visceral/transmissão , Phlebotomus/efeitos dos fármacos , Animais , Meio Ambiente , Humanos , Índia/epidemiologia , Insetos Vetores/parasitologia , Leishmania donovani , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Phlebotomus/parasitologia , Estações do AnoRESUMO
The present study is a summary of the current level of the insecticide resistance to selected organophosphates, pyrethroids, and neonicotinoids in seven Indian field populations of Bemisia tabaci genetic groups Asia-I, Asia-II-1, and Asia-II-7. Susceptibility of these populations was varied with Asia-II-7 being the most susceptible, while Asia-I and Asia-II-1 populations were showing significant resistance to these insecticides. The variability of the LC50 values was 7x for imidacloprid and thiamethoxam, 5x for monocrotophos and 3x for cypermethrin among the Asia-I, while, they were 7x for cypermethrin, 6x for deltamethrin and 5x for imidacloprid within the Asia-II-1 populations. When compared with the most susceptible, PUSA population (Asia-II-7), a substantial increase in resistant ratios was observed in both the populations of Asia-I and Asia-II-1. Comparative analysis during 2010-13 revealed a decline in susceptibility in Asia-I and Asia-II-1 populations of B. tabaci to the tested organophosphate, pyrethroid, and neonicotinoid insecticides. Evidence of potential control failure was detected using probit analysis estimates for cypermethrin, deltamethrin, monocrotophos and imidacloprid. Our results update resistance status of B. tabaci in India. The implications of insecticide resistance management of B. tabaci on Indian subcontinent are discussed.
Assuntos
Genes de Insetos , Hemípteros/efeitos dos fármacos , Hemípteros/genética , Resistência a Inseticidas , Inseticidas/farmacologia , Animais , Hemípteros/metabolismo , Índia , Fosfatos/metabolismo , Piretrinas/farmacologiaRESUMO
Two new species of leafhoppers, Hishimonus nauniensis sp. nov. (from Himachal Pradesh: Solan) and Hishimonus pantanagarensis sp. nov. (from Uttarakhand: Pant Nagar) from India, are described and illustrated. Also, the species Hishimonus thapai Virakatamath and Mohan, previously known only from Nepal is illustrated with a new record for India.
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Hemípteros/classificação , Distribuição Animal , Estruturas Animais/anatomia & histologia , Estruturas Animais/crescimento & desenvolvimento , Animais , Tamanho Corporal , Feminino , Hemípteros/anatomia & histologia , Hemípteros/crescimento & desenvolvimento , Índia , Masculino , Nepal , Tamanho do ÓrgãoRESUMO
INTRODUCTION: In the absence of capsular support, anterior chamber intraocular lens (IOL), iris fixated IOL and sutured scleral fixated intraocular lens (SFIOL) implantation have been performed for many years. Recently sutureless glued SFIOL have been used as a primary or secondary procedure to correct aphakia. In this study we have used sutureless and glueless technique of SFIOL implantation. METHODOLOGY: An interventional case series was conducted. Aphakic patients without capsular support, sub-luxated lens (>180°), dislocated lens and dislocated IOL were the inclusion criteria. The patients with hazy cornea, non-dilating pupil, macular scar and glaucoma were not enrolled in the study. RESULTS: Of 62 eyes who completed 1 month follow- up, 48 were men and 14 women. There was a significant improvement in uncorrected distance visual acuity after surgery (p less than 0.001). One month postoperative best corrected distance visual acuity was 6/18 or better in 45 eyes (72.6%). The common early postoperative complications were hypotony, corneal edema. No serious complications such as endophthalmitis and retinal detachment were seen. CONCLUSION: Our technique of sutureless and glueless SFIOL implantation showed good visual outcome in the absence of serious complications. SFIOL will be the only choice in eyes that have anatomic contraindications like non constricting pupil, large sectoral iridectomy and peripheral anterior synechia in which other types of lens are not suitable.
Assuntos
Câmara Anterior , Implante de Lente Intraocular/efeitos adversos , Lentes Intraoculares/efeitos adversos , Complicações Pós-Operatórias/etiologia , Esclera , Afacia Pós-Catarata , Feminino , Humanos , Implante de Lente Intraocular/métodos , Masculino , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a phloem-feeding, economically important pest of crops worldwide. In addition to direct damage, it also vectors a number of plant viruses belonging to the family Geminiviridae. Its populations differ biologically with respect to insecticide resistance, virus transmission and host range. Therefore, understanding genetic variation among populations is important for management. We sequenced 850 bp of the mitochondrial COI (mtCOI) gene from B. tabaci populations surveyed across India. BLAST analysis of the mtCOI sequences generated in this study with sequences from the mtCOI dataset showed the presence of one invasive group, MEAM1, and eight other groups of B. tabaci in India. mtCOI sequence analyses showed the presence of Asia I, Asia I-India, Asia II-1, Asia II-5, Asia II-7, Asia II-8, and Asia II-11 genetic groups. We also found China-3 in a field in Birbhum district, West Bengal, India, suggesting a role of anthropogenic activities in the distribution of B. tabaci. Interestingly, more than one genetic group was found coexisting in the same field.