Les nématodes à galle du genre Meloidogyne sont des parasites telluriques provoquant de graves pertes agricoles dans presque tous les systèmes de culture des plantes, et en particulier affectent la production de riz (Oryza sativa L.) dans toutes les régions cultivées. Ces parasites biotrophes obligatoires établissent une interaction compatible avec leur hôte grâce à des effecteurs protéiques produits par leurs glandes œsophagiennes et sécrétés dans la cellule végétale à travers leur stylet. L'objectif de ce travail était d'identifier parmi les protéines sécrétées celles qui jouent un rôle dans la virulence du nématode. L'interaction compatible entre la variété de riz Nipponbare et deux espèces de RKN (Meloidogyne incognita et Meloidogyne graminicola) a été choisie comme modèle et décryptée par des approches d'histologie et de transcriptomique. Trois nouvelles protéines de M. incognita spécifiquement exprimées dans les phases précoces de l'interaction ont été identifiées. Les deux gènes Mi-SP1 (Minc17980) et Mi-SP5 (Minc14137) sont exprimés dans les glandes subventrales alors que Mi-SP4 (Minc16281) s'exprime dans la glande dorsale de la larve parasitaire dite « juvénile au stade 2 (J2) ». Mi-SP1 et Mi-SP4 sont des gènes pionniers (sans homologue dans les bases de données publiques) et Mi-SP5 code potentiellement pour une déstabilase, de la famille des lysozymes. Les deux protéines pionnières Mi-SP1 et Mi-SP4 sont adressées dans le noyau et le cytoplasme de cellules de tabac après expression hétérologue en fusion avec la protéine GFP. La protéine Mi-SP5 exprimée en fusion GFP dans les cellules épidermiques d'oignon est localisée dans la paroi cellulaire. L'atténuation de l'expression des trois gènes Mi-SPs chez les J2s, induite par l'absorption de petits ARN interférants (siRNA), entraine une baisse significative de la reproduction du nématode chez le riz. De plus, la protéine Mi-SP1 permet de réprimer les défenses basales de la plante induites par le facteur bactérien flg22, telles que la production de composés réactifs d'oxygène. L'expression dans le riz de micro-ARNs artificiels (amiRNA) définis pour spécifiquement éteindre l'expression de Mi-SP5 entraine également une baisse significative de la reproduction du nématode chez le riz. L'analyse fonctionnelle de Mi-SP1 et de Mi-SP5 a montré que ces deux protéines sont capables de jouer un rôle important dans l'interaction compatible riz-nématode. / Root-knot nematode, Meloidogyne sp., are telluric pests causing severe agricultural lost in almost all plants growing system including cereals. These obligate biotrophic parasites affect the rice production in all cultivated countries. Meloidogyne incognita establishes a compatible interaction with the plant host thanks to effectors produced by esophageal glands and secreted in the plant cell through the stylet. The objective of this work was to identify secreted proteins involved in the virulence of the nematode. The compatible interaction between rice variety Nipponbare and two species of RKN (Meloidogyne incognita and Meloidogyne graminicola) was chosen as a model and decrypted by histology and transcriptomic approaches. Three new proteins of M. incognita which were specifically expressed during early stages of interaction have been identified. Two genes Mi-SP1 (Minc17980) and Mi-SP4 (Minc16281) were pioneers with no homolog in databanks and Mi-SP5 (Minc14137) encodes for a putative destabilase, belongs to lysozyme family. Mi-SP1 and Mi-SP5 were expressed in the subventral glands whereas Mi-SP4 was expressed in the dorsal gland of the parasitic larva called “juvenile stage 2 (J2)". The two pioneer proteins Mi-SP1 and Mi-SP4 were localized in both the nucleus and the cytoplasm of tobacco cell after heterologous expression whereas Mi-SP5 was located in the onion epidermal cell wall. The silencing of three effectors by soaking approach using siRNAs leads to a significant reduction in nematode reproduction in rice. In addition, Mi-SP1 can suppress the plant defences PTI triggered by the PAMP flg22. Knock-down Mi-SP5 by host-delivered RNAi also causes a significant reduction in nematode reproduction in rice. The functional analysis of Mi-SP1 and Mi-SP5 showed that these two effectors are able to play important roles in rice-nematode interaction.

Made available in DSpace on 2014-07-29T16:24:22Z (GMT). No. of bitstreams: 1 DISSERTACAO FERNANDO G DE ARAUJO.pdf: 269926 bytes, checksum: e1a81ca1614ac2d8e004f83f9641d998 (MD5) Previous issue date: 2009-11-17 / Soybean cyst nematode, Heterodera glycines Ichinohe, is one of the major phytopathological problems affecting the soybean, in the major producing countries of this legume. Eradication of H. glycines from contaminated areas is practically impossible. Thus, the soybean growers have to adapt control measures in order to maintain low population levels. The main control measures are crop rotation, soil management and use of resistant cultivars. With the purpose to evaluate the development of H. glycines and male/female ratio on soybean resistant and susceptible cultivars two trials were conducted during the years 2007 and 2008. Susceptible cultivars BRS Aracu, BRSGO Jatai, BRSGO Luziania, BRS Favorita RR, BRS Valiosa RR, BRS Silvania RR and resistant cultivars BRSGO Ipameri e BRSGO Chapadoes were evaluated. Soybean seedlings were transplanted to pots containing naturally infested soil with H. glycines, race 14, and, 10 days later, part of the plants remained in the pots to evaluate the females, while the other part was removed for hydroponic growth to evaluate the males. In the trial of 2008 the plants were divided in three lots, with the last one having the roots stained to count the juveniles and to evaluate survival rate. Resistant cultivars always maintained a small number of females and males, except for cultivar BRSGO Ipameri that had high countings of males. Only cultivars BRS Favorita RR and BRS Silvania RR had the expected sex ratio 1:1. All other susceptible cultivars had greater number of males than females, with ratios varying from 5:1 to 11:1. Survival rate was nil for both resistant cultivars, and varied from 6.75% to 35.00% for the susceptible cultivars. Regarding the number of cysts in 100 cc of soil the only cultivar that differed significantly from the others was the BRSGO Jatai on experiment conducted in 2007. The number of eggs per cyst varied, in both experiments, and the resistance did not influence the averages found / nematóide de cisto da soja, Heterodera glycines, é um dos principais problemas fitossanitários que afeta a cultura da soja nos principais países produtores desta oleaginosa. A erradicação do H. glycines nas áreas contaminadas é praticamente impossível. Dessa forma, o produtor deve adotar práticas culturais com o intuito de manter baixos os níveis populacionais. As principais medidas de controle são a rotação de culturas, o manejo de solo e a utilização de cultivares resistentes. Com o objetivo de avaliar o desenvolvimento de H. glycines e a proporção macho:fêmea em cultivares de soja resistentes e suscetíveis foram realizados dois experimentos, sendo um em 2007 e outro em 2008. Avaliou-se as cultivares suscetíveis BRSGO Araçu, BRSGO Jataí, BRSGO Luziânia, BRS Favorita RR, BRS Valiosa RR, BRS Silvânia RR e as cultivares resistentes BRSGO Ipameri e BRSGO Chapadões. Plântulas pré-germinadas de soja foram transferidas para vasos com solo naturalmente infestado com H. glycines, raça 14, e, após dez dias, parte das plantas permaneceram nos vasos para avaliação de fêmeas e cistos no solo, e outra parte foi utilizada para instalação do sistema hidropônico para avaliação de machos. No ensaio conduzido em 2008, as plantas foram divididas em três partes sendo a última destinada à coloração de raízes para contagem de juvenis e avaliação da taxa de sobrevivência. As cultivares resistentes sempre mantiveram baixo número de fêmeas e machos com exceção da cultivar BRSGO Ipameri que obteve altos índices de machos. Somente as cultivares BRS Favorita RR e BRS Silvânia RR produziram uma relação macho/fêmea de aproximadamente 1:1. As demais cultivares suscetíveis apresentaram número de machos maior que o de fêmeas com proporções variando de 5:1 a 11:1. A taxa de sobrevivência foi nula para ambas as cultivares resistentes e variou de 6,75% a 35,00% para as cultivares suscetíveis. Com relação ao número de cistos em 100 cm³ de solo, a única cultivar que diferiu significativamente das demais foi a BRSGO Jataí no experimento realizado em 2007. O número de ovos por cisto variou bastante, em ambos os experimentos, sendo que a resistência não influenciou as médias encontradas

Made available in DSpace on 2016-04-28T20:15:29Z (GMT). No. of bitstreams: 1 2008- Daniela de Oliveira Franco Acuna.pdf: 2634530 bytes, checksum: 5433beffd9a4776e67f732e78762b6bb (MD5) Previous issue date: 2008-02-19 / The snail, Achatina fulica (Giant African snail), is registered in almost all Brazilian territory, and may act as an intermediate host of parasites of the man. This study aimed to describe the morphology of the nematode larvae found in A. fulica, through light (LM) and scanning electron microscopy (SEM), and to relate the prevalence of infection to the collection places of the snails and to the antropic activity. Sixty snails were collected from April/2006 to April/2007, in the Mesquita City, RJ, transferred to laboratory, kept in terraria, measured and dissected to verify the infection, distribution of the cysts in tissues of the snails. The cysts were collected and the larvae were separated. The material thus obtained was used to LM and SEM. The occurrence of A. fulica in the less conserved area was greater than that in the more conserved area. However, there was not a significant relation between the intensity of infection and the collection places and the antropic activity. Fourteen snails (23%) were infected and the biggest number of cysts/snail verified was 130 and the minor was 1, in snails that with 106 and 55mm of shell length, respectively. A strong correlation was observed between the size of the snail and the number of cysts and there was a greater concentration of these in the pallial cavity, 87%. Of this total, 49% were in the area more vascularized and 38% in the less vascularized area of this cavity. The cysts presented spherical shape, light pink colored and measured 0.97-1.57mm in diameter and with an unique larva involved by an amorphous material. Two hundred twenty two encysted larvae were recovered, 30 larvae were used to the morphological studies. The length of the larvae varied of 2.57-5.8mm, being classified in: small - until 3.5mm; medium - of 3.53-4.5mm and great - above of 4.52mm. The average length of the larvae in the three groups was of 2.85; 3.87 and 5.23mm, respectively, and the total average of 4.2mm. The larvae presented white cuticle, shining, transversally striated and the lateral line extending to the posterior extremity of the body. In the anterior end, it is observed mouth provided with three lips, with amphids and papillae, followed by a muscular oesophagus with average length of 0.61mm, ending in an esophagus bulb. The nervous ring it was observed in the medium third of the oesophagus and the intestine ends in an opening located next to the posterior end. The tail had a length varying of 0.15-0.42mm, is located after this opening may present two types of endings: abrupt or that it sharpens gradually. The difference in the ending of the tail may suggest sexual dimorphism, although not to have been observed reproductive structures. The identification to the specific level was not possible, this pointed to the necessity of continuation of this study. The expressive population density of A.fulica observed in the conserved area less it confirms the influence of the antropic action on the dispersion of this snail. / O molusco Achatina fulica (Caramujo gigante africano) ? registrado em quase todo territ?rio nacional, havendo a possibilidade de atuar como hospedeiro de parasitos do homem. Este estudo objetivou descrever a morfologia dos est?gios larvais de nemat?ides encontrados em A. fulica, atrav?s de microscopia de luz e eletr?nica de varredura, e relacionar a preval?ncia de infec??o aos locais de coleta dos moluscos e ? atividade antr?pica. Sessenta moluscos foram coletados de abril de 2006 a abril de 2007, no munic?pio de Mesquita, RJ, transferidos para o laborat?rio, mantidos em terr?rios, medidos e dissecados para verifica??o da infec??o, distribui??o dos cistos pelos tecidos do molusco, coleta dos cistos e obten??o das larvas. Procedeu-se a fixa??o e o processamento para microscopia de luz e eletr?nica de varredura. A ocorr?ncia de A. fulica na ?rea menos conservada foi maior que na ?rea mais conservada. Por?m n?o houve uma rela??o significativa entre a intensidade de infec??o aos locais de coleta e ? atividade antr?pica. Quatorze moluscos (23%) estavam infectados e o maior n?mero de cistos verificado por molusco foi de 130 e o menor de 01, em moluscos que apresentavam 106 e 55mm de comprimento de concha, respectivamente. Foi observada uma forte correla??o entre o tamanho do molusco e o n?mero de cistos encontrados e maior concentra??o destes na cavidade paleal, 87%. Desse total, 49% estavam na ?rea mais vascularizada e 38% na menos vascularizada da cavidade. Os cistos apresentavam formato esf?rico, colora??o rosada e mediam de 0,97 a 1,57mm de di?metro e na maioria das vezes com uma ?nica larva que se encontrava envolvida por um material amorfo. Foram recuperadas 222 larvas encistadas. Destas, 30 foram utilizadas no estudo morfol?gico. O comprimento das larvas variou de 2,57 a 5,8mm, sendo classificados em: pequeno at? 3,5mm; m?dio de 3,53 a 4,5mm e grande - acima de 4,52mm. O comprimento m?dio das larvas nos tr?s grupos foi de 2,85; 3,87 e 5,23mm, respectivamente, e a m?dia total de 4,2mm. As larvas apresentavam cut?cula branca, brilhante e estriada no sentido transversal e linha lateral at? a extremidade posterior do corpo. Na extremidade anterior, observa-se a boca provida de tr?s l?bios, com anf?dios e papilas, seguida por um es?fago musculoso com comprimento m?dio de 0,61mm, terminando em um bulbo esofagiano. O anel nervoso foi observado no ter?o m?dio do es?fago e o intestino termina em uma abertura localizada pr?ximo ? extremidade posterior. A cauda, com comprimento variando de 0,15 a 0,42mm, se forma a partir desta abertura e apresenta dois tipos de termina??es: abrupta ou que afina gradativamente. A diferen?a na termina??o da cauda pode sugerir dimorfismo sexual, apesar de n?o terem sido observados prim?rdios de ?rg?os reprodutores. Com os resultados obtidos n?o foi poss?vel a identifica??o ao n?vel espec?fico, havendo dessa forma a necessidade de continua??o deste estudo. A densidade populacional expressiva de A. fulica observada na ?rea menos conservada confirma a influ?ncia da a??o antr?pica sobre a dispers?o deste molusco.

Estudaram-se a estruturas taxionômica e trófica e determinaram-se os índices de maturidade de comunidades de nematóides ocorrentes em áreas contíguas de mata natural preservada e de culturas perenes (bananeira, citros, pessegueiro) e anuais (milho x leguminosas) em rotação em Piracicaba (SP). Três amostras compostas de solo (volume aproximado de um litro) foram coletadas de cada área em duas épocas (\"seca\", agosto/2004 e \"chuvosa\", março/2005) e profundidades (0-15 e 15-30 cm). Para cada amostra, extraíram-se os nematóides de volume de 500 ml pela técnica de peneiramento combinado com flotação em centrífuga com solução de sacarose, sendo depois fixados em formalina, contados e infiltrados com glicerina. Para o cálculo das abundâncias, indicadores de diversidade e os índices de maturidade, 150 exemplares foram coletados ao acaso por amostra e identificados ao nível de gênero. Assinalaramse 61 gêneros, filiados a 32 famílias. As abundâncias totais foram maiores à menor profundidade, nas duas épocas de coleta. Os maiores valores de número e riqueza de gêneros, à menor profundidade, ocorreram na mata; à maior profundidade, tal se deu na área de anuais e de citros, nas primeira e segunda épocas de coleta, respectivamente. Os índices de diversidade de Shannon- Weaver e de Simpson e respectivos índices de eqüitatividade foram menos eficientes que o número e a riqueza de gêneros na discriminação dos sistemas estudados. Os parasitos de plantas constituíram o grupo trófico prevalente em todos os sistemas avaliados, nas duas épocas e profundidades de coleta, seguidos no geral pelos bacteriófagos. Todavia, não foi possível separar cultivos anuais de perenes com base nas abundâncias de parasitos de plantas. Os valores de T (diversidade trófica) não se prestaram à clara distinção entre os cinco sistemas, mas os de MI, mMI e PPI indicaram menor ocorrência de distúrbios na mata e no pomar cítrico em relação às demais áreas cultivadas. A herbivoria foi o canal prevalente de decomposição nos diferentes sistemas, exceto no pomar de citros. Dentre os microbiófagos (FF/BF), a predominância foi da via bacteriana. / The taxonomic structure, trophic structure and maturity index of nematode communities occurring in neighbouring areas of native forest and four different agroecosystems were studied in Piracicaba, São Paulo State, Brazil. Agroecosystems were three perennial (banana, citrus and peach orchards) and one annual (corn, rotating with leguminous plants or followed by fallow) crops. Three soil samples (one liter approximately) were collected per area at two different times (August/2004 and March/2005) and depths (0-15 and 15-30 cm). Nematodes were extracted from 500 cc of soil by a sieving and sugar flotation technique, fixed with formalin, counted and infiltrated with glycerin. Subsequently, 150 specimens randomly picked were identified to the genus level. A total number of 61 genera affiliated to 32 families was recorded. Total abundance was higher at lower depth irrespective of the sampling time. At depth 0-15 cm, higher values of genus number and genus richness were calculated for the forest in both sampling times; at 15-30 cm, the highest values were assigned to annual crop and citrus areas, respectively. Shannon- Weaver and Simpson indexes were less efficient than genus number and genus richness in discriminating the studied ecosystems. Plant-parasitic nematodes prevailed as a trophic group in all systems at both sampling times and depths, the bacterivorous usually ranking second position. However, the data of relative abundance of phytonematodes did not allow annual and perennial crop areas to be statistically differentiated. The studied systems also were not clearly distinguished with basis on the T values, but according to the MI, mMI and PPI data the forest and the citrus orchard were the less disturbed areas. Herbivory was the predominant decomposition channel in all systems, with exception of the citrus orchard. In terms of microbial decomposition (FF/BF), the process was usually dominated by bacteria.

Orientador: Mário Massayuki Inomoto / Resumo: Amostras de solo e raízes foram coletadas em plantações de café no estado de São Paulo, com o objetivo de determinar a importância e a ocorrência dos nematóides das lesões na cultura do café. A espécie de Pratylenchus mais freqüente foi P. brachyurus (solo: 13,2 %; raízes: 18,3 %), mas geralmente em baixas densidades. O nematóide das lesões do café, P. coffeae, ocorreu em 5,1 % das amostras de raízes, mas em altas densidades e causando mais danos do que a primeira espécie. Outra espécie, P. vulnus, foi encontrada somente em uma localidade. Este é o primeiro relato de P. vulnus em café. Outros fitonematóides identificados nas amostras foram: Xiphinema brevicolle, Xiphinema sp., Paratrichodorus minor, Paratrichodorus sp., ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Soil and root samples were collected from coffee plantations in the state of São Paulo, in order to determine the occurrence and importance of lesion nematodes in coffee culture. The most frequent species of Pratylenchus was P. brachyurus (soil: 13,2 %; roots: 18,3 %), but generally in low densities. The coffee lesion nematode, P. coffeae, occurred in 5,1 % of root samples, but in higher densities and causing more damage than the former species. Other species, P. vulnus, was found in one single locality. This is the first report of P. vulnus on coffee. Other phytonematodes identified in the survey were: Xiphinema brevicolle, Xiphinema sp., Paratrichodorus minor, Paratrichodorus sp., Tylenchorhynchus sp., Helicotylenchus dihystera, H. californicus, H. erythrinae, Helicotylenchus sp., Scutellonema sp., Rotylenchulus reniformis, Meloidogyne exigua, M. incognita, M. coffeicola, Meloidogyne sp, Criconemella onoensis, C. ornata, C. sphaerocephala, Criconemella sp., Hemicriconemoides strictathecatus, and Paratylenchus sp. ...(Complete abstract, click electronic access below) / Doutor

Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Examines the distribution, biology and ecology of the Anguina-type nematodes and the bacterium Rathayibacter toxicus which are associated with flood plain staggers, a disease similar to annual ryegrass toxicity. Possible pasture and livestock management options to control the disease are discussed, as is biological control using the fungus Dilophospora alopecuri. / Thesis (Ph.D.)--University of Adelaide, School of Agriculture and Wine, Discipline of Plant and Pest Science, 2003?

Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2006.

Root-lesion nematodes, Pratylenchus spp., are serious pathogens of potato plants worldwide. Several management practices can control Pratylenchus spp.; however, they all have shown some limitations. Therefore, environmentally-safe, low-cost, and effective control strategies are needed as possible alternative to currently used strategies. This thesis was designed to assess if liquid hog manure (LHM) holds such potential. The objectives of this thesis were to determine: (i) the prevalence and identity of species of Pratylenchus spp. in Manitoba potato fields, (ii) if short-chain volatile fatty acids (VFA) in LHM are the constituents responsible for the manure’s toxicity to Pratylenchus spp. using solution exposure experiments (iii) the effectiveness of LHM in killing Pratylenchus spp. in soil, and (iv) the impact of LHM on nematode communities. Pratylenchus spp. were detected in 39% of 31 potato fields surveyed in Manitoba with population densities ranging, for positive fields, from 45 to 631 nematodes kg-1 fresh soil. Morphometrics of female nematodes and molecular diagnosis (using species-specific PCR primers) showed that the species of Pratylenchus present in the potato fields to be P. neglectus. Potato, cv. Russet Burbank, showed to be a poor host to two populations of Pratylenchus spp. from Manitoba potato fields. Accordingly, P. neglectus does not seem to be a limitation to potato production in Manitoba; thus, P. penetrans, the most widely spread and damaging species to potato was used in the successive studies of assessing the use of LHM to control Pratylenchus spp. in potato fields. VFA (acetic, propionic, n-butyric, isobutyric, n-valeric, isovaleric, and n-caproic acids) accounted for the majority of the lethal effect of LHM to P. penetrans under acidic conditions. VFA in LHM killed Pratylenchus spp. in soil and acidification seemed to enhance its ability when VFA concentration in the manure is low. LHM did not act as a soil fumigant eliminating soil trophic interactions but increased bottom-up food web interactions. VFA in LHM persisted in the soil for four days with biological degradation being their mode of loss. In conclusion, LHM is potentially an effective and low-cost strategy to control Pratylenchus spp. and its efficacy can be improved by acidification. / October 2009

Previous studies have indicated that the promoters of the human filarial parasite Brugia malayi are unusual in that they do not exhibit the CAAT or TATAA sequences usually found in the core domains of promoters of most eukaryotic organisms. Analysis of the promoters of the ribosomal proteins showed that the region flanking the splice leader (SL) addition site plays an important role in transcription and may function as the core promoter domain in B. malayi. To test the hypothesis that the SL addition domain is the most important essential region of the ribosomal protein promoters, the SL addition site of the BmRPL13 gene was replaced with the SL addition domains from other ribosomal protein genes from B. malayi. The promoter activity of the replacement constructs were tested using a transient transfection dual luciferase assay. Promoter activity with RPL13 replacement constructs was correlated with that seen in the wild type promoters, suggesting that roughly 80% of the variations seen in promoter activity among ribosomal protein promoters is due to variation in the SL core promoter domain.

The aim of this study was to determine whether functional ecdysone response elements (EcREs) exist within the genome of Brugia malayi, a parasitic nematode that causes lymphatic filariasis. The hypothesis that EcREs exist in B. malayi stemmed from previous demonstration of a functional ecdysone response system in B. malayi (Tzertzinis et al., 2010). Real-time PCR (qPCR) experiments were conducted to measure gene expression levels for twelve genes proximal to five putative EcREs in 20-OH ecdysone treated and untreated B. malayi embryos. Seven genes showed consistent upregulation with 20-OH ecdysone treatment. Each of the five putative EcREs had at least one proximal gene consistently upregulated, suggesting that all five might be functional EcREs. One of the genes consistently upregulated in the qPCR experiments, Bm1_48650, codes for a MIZ zinc finger family protein, a likely transcription factor. Transgenic ecdysone induction assay experiments were conducted using embryos transiently transfected with a reporter construct driven by the EcRE-containing promoter of Bm1_48650. Significantly higher mean reporter gene activity (~3.5-fold) was seen in 20-OH ecdysone treated versus untreated embryos. In another set of transgenic ecdysone induction assays, the EcRE motif in the Bm1_48650 promoter was completely mutated, and this construct was tested in 20-OH ecdysone treated and untreated embryos. The mean reporter gene activity for the treated and untreated embryos transfected with the mutant constructs did not differ significantly from the untreated embryos transfected with the native EcREcontaining promoter construct. These results showed that the EcRE in the promoter of Bm1_48650 is necessary for regulating gene expression in response to 20-OH ecdysone. This study substantiates previously discovered evidence of a functional ecdysone response system in B. malayi, which could potentially serve as a target for drug discovery for lymphatic filariasis.