Bancroftian Filariasis Research Articles

Designing of a chimeric multiepitope vaccine against bancroftian lymphatic filariasis through immunoinformatics approaches

The filarial worms of Wuchereria bancrofti are the primary cause of lymphatic filariasis (LF), a mosquito-borne disease among the neglected tropical parasitic diseases. Considering the global endemic consequences of the disease, there is a need to develop a successful vaccine candidate against LF. Using advanced immunoinformatics approaches, we designed two multiepitope vaccines targeting W. bancrofti’s glutathione S-transferase and thioredoxin. Therefore, we predicted several MHC-1, MHC-2, and B-cell epitopes from these proteins and mapped two vaccine candidates (V1 and V2). The vaccines were subsequently employed for physicochemical analysis, structural prediction and validation, docking and normal mode analysis, codon optimization, and immune simulation. The selected MHC-1, MHC-2, and B-cell epitopes were antigenic without allergenicity or toxicity. The designed vaccines were expected to be soluble, stable proteins under physiological conditions. Compared to V2, V1’s secondary and tertiary structures were simultaneously favorable, with Ramachandran plot analysis revealing 95.6% residues in favored areas. Subsequently, the molecular docking analysis indicated that the V1 had a high binding affinity for the TLR-2, TLR-4 and TLR-5, as suggested by the docking scores of -1248.7, -1038.5 and -1562.8, respectively. The NMA of these complexes further indicated their structural flexibility. Molecular dynamics simulations of V1-TLR complexes revealed V1-TLR-4 as the most stable, with the lowest free energy and minimal fluctuations, indicating the strongest binding affinity. The results of the codon optimization showed high levels of expression, with a favorable CAI score (<1.0). A three-dose vaccination analysis showed significant and persistent immunological responses, including adaptive and innate immune responses. The findings emphasize the potential of the V1 against W. bancrofti, but further validation is required through in vitro, in vivo, and clinical trials.

Detection of Wuchereria bancrofti infection in mosquitoes in areas co-endemic with Brugia malayi in Balasore district, Odisha, India

Lymphatic filariasis (LF) is a crippling and disfiguring parasitic condition. India accounts for 55% of the world’s LF burden. The filarial parasite Wuchereria bancrofti is known to cause 99.4% of the cases while, Brugia malayi accounts for 0.6% of the issue occurring mainly in some pockets of Odisha and Kerala states. The Balasore (Baleswar) district of Odisha has been a known focus of B. malayi transmission. We employed molecular xenomonitoring to detect filarial parasite DNA in vectors. In six selected villages, Gravid traps were used to collect Culex mosquitoes and hand catch method using aspirators was followed for collection of mansonioides. A total of 2903 mosquitoes comprising of Cx. quinquefasciatus (n = 2611; 89.94%), Cx. tritaeniorhynchus (n = 100; 3.44%), Mansonia annuliferea (n = 139; 4.78%) and Mansonia uniformis (n = 53; 1.82%) were collected from six endemic villages. The species wise mosquitoes were made into 118 pools, each with a maximum of 25 mosquitoes, dried and transported to the laboratory at VCRC, Puducherry. The mosquito pools were subjected to parasite DNA extraction, followed by Real-time PCR using LDR and HhaI probes to detect W. bancrofti and B. malayi infections, respectively. Seven pools (6.66%) of Cx. quinquefasciatus, showed infection with only W. bancrofti while none of the pools of other mosquito species showed infection with either W. bancrofti or B. malayi. Although the study area is endemic to B. malayi, none of the vectors of B. malayi was found with parasite infection. This study highlights the ongoing transmission of bancroftian filariasis in the study villages of Balasore district of Odisha and its implications for evaluating LF elimination programme.

Lethal and sublethal effects of spinosad on dengue vector mosquito, Aedes albopictus and bancroftian filariasis vector mosquito, Culex pipiens pallens.

Aedes albopictu and Culex pipiens pallens are important vectors of many viruses and have had resistance to chemical pesticide. Spinosad is a selective biological insecticide to control urban mosquito. The aim of this study was to reveal the sublethal effects of spinosad on mosquito and provide reference basis for integrated mosquito management. The toxicity of spinosad against Ae. albopictus and Cx. pipiens pallens were determined under laboratory conditions by exposing early third-instar larvae to different concentrations. The LC50 values of spinosad to Ae. albopictus and Cx. Pipiens pallens larvaes were 4.44×10-3 mg∙L-1 and 1.93×10-3 mg∙L-1 respectively after 72 h exposure. Spinosad at sublethal concentrations has many negative effects on Ae. albopictus and Cx. Pipiens pallens larval, pupae, adult and offspring eggs, including significantly reduced their larvae pupation rate by 51.37% and 58.47%, significantly prolonged pupae length by 21.43% and 16.18%, reduced female wing-spans by 20.19% and 14.89%, reduced male wing-spans by 3.84% and 7.54%, reduced female weight by 29.04% and 31.52%, reduced male weight by 7.47% and 9.07%, reduced female and male ratio by 51.98% and 45.21%, reduced individual egg-laying amount by 15.73% and 35.51%, in addition, offspring egg hatchability were dramatically decreased by 25.71% and 34.04%, egg periods were significantly prolonged by 14.42% and 62.82% respectively. No significant effect on larval period, pupae emergence rate, female bite rates were observed. These results suggest that spinosad might affect pest population dynamics significantly and is fairly expected to be a candidate biological pesticide for mosquito control.

Pharmacokinetics of Moxidectin combined with Albendazole or Albendazole plus Diethylcarbamazine for Bancroftian Filariasis.

Moxidectin (MOX) is a milbemycin endectocide recently approved by the U.S. FDA for the treatment of onchocerciasis in persons at least 12 years of age. MOX has been shown to have a good safety profile in recent clinical trials. The efficacy of MOX for the treatment of lymphatic filariasis (LF) and its potential use in mass drug administration protocols for the elimination of LF is currently under evaluation. In the context of a clinical trial, we investigated the pharmacokinetics and drug interactions of a combination of MOX plus albendazole (ALB) with or without diethylcarbamazine (DEC) compared to ivermectin (IVM) plus ALB with or without DEC in the following four different treatment arms: (I) IVM (0.2mg/kg) plus DEC (6 mg/kg) and ALB (400mg); (II) IVM plus ALB; (III) MOX (8 mg) plus DEC and ALB; and (IV) MOX plus ALB. Drug concentrations were determined using validated liquid chromatography-mass spectrometric methods. Pharmacokinetic parameters were determined using standard non-compartmental analysis methods. Statistical analysis was performed using JMP software. Fifty-eight of 164 study participants (53 men and five women) were included with ages ranging from 18 to 63 yrs (mean = 37). MOX apparent oral clearance (Cl/F) ranged from 0.7 to 10.8 L/hr with Cmax values ranging from 20.8 to 314.5 ng/mL. The mean (range) area under the curve (AUC)0-∞ for MOX, 3405 ng*hr/mL (742-11376), and IVM 1906 ng*hr/mL (692-5900), varied over a ~15.3 and ~8.5-fold range, respectively. The geometric mean ratio for Cmax, AUC0-t, and AUC0-∞ were within the no-drug interaction range of 80-125% for all drugs. This indicates that the addition of MOX to ALB alone or ALB plus DEC for LF therapy did not alter the drug exposure of co-administered drugs compared to IVM combinations. Clinical Trial Registration: NCT04410406, https://clinicaltrials.gov/.

Evaluation of the evolutionary genetics and population structure of Culex pipiens pallens in Shandong province, China based on knockdown resistance (kdr) mutations and the mtDNA-COI gene

BackgroundMosquitoes are important vectors for a range of diseases, contributing to high rates of morbidity and mortality in the human population. Culex pipiens pallens is dominant species of Culex mosquito in northern China and a major vector for both West Nile virus and Bancroftian filariasis. Insecticide application were largely applied to control the mosquito-mediated spread of these diseases, contributing to increasing rates of resistance in the mosquito population. The voltage-gated sodium channel (Vgsc) gene is the target site of pyrethroids, and mutations in this gene cause knockdown resistance (kdr). While these kdr mutations are known to be critical to pyrethroid resistance, their evolutionary origins remain poorly understood. Clarifying the origins of these mutations is potential to guide further vector control and disease prevention efforts. Accordingly, the present study was designed to study the evolutionary genetics of kdr mutations and their association with the population structure of Cx. p. pallens in Shandong province, China.MethodsAdult Culex females were collected from Shandong province and subjected to morphological identification under a dissection microscope. Genomic DNA were extracted from the collected mosquitoes, the Vgsc gene were amplified via PCR and sequenced to assess kdr allele frequencies, intron polymorphisms, and kdr codon evolution. In addition, population genetic diversity and related population characteristics were assessed by amplifying and sequencing the mitochondrial cytochrome C oxidase I (COI) gene.ResultsTotally, 263 Cx. p. pallens specimens were used for DNA barcoding and sequencing analyses to assess kdr allele frequencies in nine Culex populations. The kdr codon L1014 in the Vgsc gene identified two non-synonymous mutations (L1014F and L1014S) in the analyzed population. These mutations were present in the eastern hilly area and west plain region of Shandong Province. However, only L1014F mutation was detected in the southern mountainous area and Dongying city of Shandong Province, where the mutation frequency was low. Compared to other cities, population in Qingdao revealed significant genetic differentiation. Spatial kdr mutation patterns are likely attributable to some combination of prolonged insecticide-mediated selection coupled with the genetic isolation of these mosquito populations.ConclusionsThese data suggest that multiple kdr alleles associated with insecticide resistance are present within the Cx. p. pallens populations of Shandong Province, China. The geographical distributions of kdr mutations in this province are likely that the result of prolonged and extensive insecticide application in agricultural contexts together with frequent mosquito population migrations. In contrast, the low-frequency kdr mutation detected in central Shandong Province populations may originate from the limited selection pressure in this area and the relative genetic isolation. Overall, the study compares the genetic patterns revealed by a functional gene with a neutral marker and demonstrates the combined impact of demographic and selection factors on population structure.

Michelia champaca seed extract: A novel mosquitocide against the filarial vector Culex quinquifasciatus

Vector control is in risk as a result of the advent of synthetic pesticide resistance. Plant derived insecticides may be used as future bio control alternatives due to their target specificity and biodegradability. The aim of the present study was to determine the efficacy of Chloroform: Methanolic extract of Michelia champaca seeds against all the larval instars of Culex quinquifasciatus, the vector of Bancroftian filariasis. Chloroform: Methanolic extract showed remarkable larvicidal properties with LC50 and LC90 values as low as 15.33 ppm and 29.77 ppm recorded against the first instar larvae after 72 hours of treatment. There was a clear dose dependent increment in the death rate, which was positively correlated with the solvent concentration and had regression coefficient values close to 1. Because the investigated non-target organisms were unaffected, the plant-based mosquitocide that was thus discovered was proved to be environment friendly. FTIR analysis revealed the presence of various functional groups in the TLC fraction such as alcohol, alkane, methyl group, amine, aromatics, carboxylic acid and ether. So, M. champaca seeds extract can be proclaimed as an outstanding larvicidal agent against Cx. quinquifasciatus, and can serve as a source of potent mosquito larvicide in the near future.

Prevalence and clinical manifestations of Bancroftian filariasis in northern Taraba State, Nigeria

Prevalence and clinical manifestations of Bancroftian filariasis in northern Taraba State, Nigeria

Characterization of a novel microfilarial antigen for diagnosis of Wuchereria bancrofti infections.

BackgroundLymphatic filariasis (LF) is a neglected tropical disease caused by the filarial nematodes Wuchereria bancrofti, Brugia malayi and Brugia timori. The Global Program to Eliminate LF uses mass drug administration (MDA) of anti-filarial drugs that clear microfilariae (Mf) from blood to interrupt transmission by mosquitos. New diagnostic tools are needed to assess the impact of MDA on bancroftian filariasis, because available serologic tests can remain positive after successful treatment.Methodology/Principal findingsWe identified Wb-bhp-1, which encodes a W. bancrofti homologue of BmR1, the B. malayi protein used in the Brugia Rapid antibody test for brugian filariasis. Wb-bhp-1 has a single exon that encodes a 16.3 kD protein (Wb-Bhp-1) with 45% amino acid identity to BmR1. Immunohistology shows that anti-Wb-Bhp-1 antibodies primarily bind to Mf. Plasma from 124 of 224 (55%) microfilaremic individuals had IgG4 antibodies to Wb-Bhp-1 by ELISA. Serologic reactivity to Wb-Bhp-1 varied widely with samples from different regions (sensitivity range 32–92%), with 77% sensitivity for 116 samples collected from microfilaremic individuals outside of sub-Saharan Africa. This variable sensitivity highlights the importance of validating new diagnostic tests for parasitic diseases with samples from different geographical regions. Individuals with higher Mf counts were more likely to have anti-Wb-Bhp-1 antibodies. Cross-reactivity was observed with a minority of plasma samples from people with onchocerciasis (17%) or loiasis (10%). We also identified, cloned and characterized BmR1 homologues from O. volvulus and L. loa that have 41% and 38% identity to BmR1, respectively. However, antibody assays with these antigens were not sensitive for onchocerciasis or loiasis.ConclusionsWb-Bhp-1 is a novel antigen that is useful for serologic diagnosis of bancroftian filariasis. Additional studies are needed to assess the value of this antigen for monitoring the success of filariasis elimination programs.

Semiannual Treatment of Albendazole Alone is Efficacious for Treatment of Lymphatic Filariasis: A Randomized Open-label Trial in Cote d'Ivoire.

Ivermectin (IVM) plus albendazole (ALB), or IA, is widely used in mass drug administration (MDA) programs that aim to eliminate lymphatic filariasis (LF) in Africa. However, IVM can cause severe adverse events in persons with heavy Loa loa infections that are common in Central Africa. ALB is safe in loiasis, but more information is needed on its efficacy for LF. This study compared the efficacy and safety of 3 years of semiannual treatment with ALB to annual IA in persons with bancroftian filariasis. Adults with Wuchereria bancrofti microfilaremia (Mf) were randomized to receive either 3 annual doses of IA (N = 52), 6 semiannual doses of ALB 400 mg (N = 45), or 6 semiannual doses of ALB 800 mg (N = 47). The primary outcome is amicrofilaremia at 36 months. IA was more effective for completely clearing Mf than ALB 400mg or ALB 800mg (79%, 95% confidence interval [CI]: 67-91; vs 48%, 95% CI: 32-66 and 57%, 95% CI: 41-73, respectively). Mean percentage reductions in Mf counts at 36 months relative to baseline tended to be greater after IA (98%, 95% CI: 88-100) than after ALB 400 mg (88%, 95% CI: 78-98) and ALB 800 mg (89%, 95% CI: 79-99) (P = .07 and P = .06, respectively). Adult worm nest numbers (assessed by ultrasound) were reduced in all treatment groups. Treatments were well tolerated. Repeated semiannual treatment with ALB is macrofilaricidal for W. bancrofti and leads to sustained reductions in Mf counts. This is a safe and effective regimen that could be used as MDA to eliminate LF in areas where ivermectin cannot be used. NCT02974049.

Molecular and Antifilarial IgG4 Detection Using the miniPCR-Duplex Lateral Flow Dipstick and BmSxp-ELISA in Myanmar Immigrant Communities

Lymphatic filariasis (LF) is an important tropical disease that affects over a billion people in more than 80 countries and approximately 40 million people are currently suffering from severe disfigurement and disability. A diagnostic tool is the principal impact factor to determine the infection status of lymphatic filariasis. The purpose of the present study was to investigate nucleic acid of Wuchereria bancrofti as well as antifilarial IgG4 in a Myanmar immigrant community living along the Moei River, a natural border between Mae Sot, Tak province Thailand and Myawaddy, Myanmar which is an endemic area of bancroftian filariasis. Blood was collected from 300 Myanmar immigrants in Mae Sot district, Tak Province. The nucleic acid of W. bancrofti was assessed in the study population using our recent published miniPCR-Duplex Lateral Flow dipstick (DLFD) platform as well as the standard PCR technique. The antifilarial IgG4 was detected in the study population using the developed ELISA which used BmSxp protein as antigen. The miniPCR-DLFD method delivered results comparable to the standard PCR technique and it enables convenient and rapid visual detection of the parasite nucleic acid. Furthermore, the ELISA using BmSxp antigen demonstrated a sensitivity, specificity, and positive and negative predictive values of 98.1%, 98.9%, 96.3%, and 99.4% respectively. The W. bancrofti nucleic acid and antifilarial IgG4 were detected in 1.6% (5/300), and 2% (6/300) of the study population, accordingly. The results of this study also revealed important epidemiological data about LF on the Thai–Myanmar border.

Filariasis diagnosed by real-time ultrasound scanning as filarial dance sign - A case report.

IntroductionLymphatic filariasis (LF) is a vector-borne illness caused due to Wuchereria bancrofti. Genital abnormalities may result from chronic inflammation of the lymphatic vessels and obstruction of the afferent vessels in Bancroftian filariasis. Doxycycline, albendazole, diethylcarbamazine, and ivermectin are among the treatments available.CaseA 16-year male presented to our OPD with a complaint of left inguinal swelling for the past 6 Months. Examination of the swelling revealed a left-sided inguinal swelling of 5 × 4 cm with intact overlying skin. A provisional diagnosis of inguinal lymph varix was made, and real-time ultrasound scanning along with FNAC was performed. Swelling resolved after conservative management.DiscussionUltrasonography can easily observe adult worms due to their size and fast movements. Using pulsed-wave doppler mode, the characteristic movement profile of adult worms in pulsed-wave doppler mode can be distinguished from other movements (e.g., blood in venous vessels).ConclusionUltrasound is the imaging modality of choice to detect filarial worms/microfilaria in the lymphatic system, which produces the classic twirling of microfilaria in the lymphatic system, known as the "filarial dance sign".

Screening of Capillaria philippinensis infection using Trichuris muris and Trichinella spiralis antigens

Background: Intestinal capillariasis is a disease caused by Capillaria philippinensis. Human infectionsbecame more prevalent in many countries including Egypt. This nematode is related to Trichinella andTrichuris species, all of them belong to Trichinelloidea superfamily. Diagnosis of intestinal capillariasismay be missed by stool examination.Objective: This study aimed to use enzyme-linked immunosorbent assay (ELISA) and immunoblotting inscreening for intestinal capillariasis as a practical and rapid diagnostic test.Subjects and Methods: Trichuris muris and Trichinella spiralis adult worms were isolated from infectedmice and crude antigens were prepared. The protein content for both adult worm extracts was determined.Human blood samples were collected from 20 capillariasis patients, 20 control individuals and 10 W.bancrofti infected patients. The study evaluated cross-reactivity between T. spiralis and T. muris antigensand sera from cases infected with C. philippinensis using ELISA and immunoblotting.Results: In ELISA, T. muris crude antigen cross reacted with 100% of capillariasis sera with 100%sensitivity and specificity and cross reacted with 10% of sera from bancroftian filariasis. T. spiralis crudeantigen cross reacted with 50% of capillariasis sera, and 9% of sera from bancroftian filariasis. Neither T.muris nor T. spiralis crude antigens reacted with sera from control group. Immunoblotting results showedthat IgG antibodies from control group didn't recognize specific proteins in T. muris antigen, while in W.bancrofti group, only one band in one sample appeared (100-135 kDa). IgG antibodies from capillariasiscases recognized multiple common protein bands (35-180 kDa). IgG antibodies from capillariasis, controland W. bancrofti sera did not recognize specific proteins in T. spiralis crude antigen.Conclusion: Antigens from T. muris and T. spiralis can be used successfully to detect infection with C.philippinensis using serum samples of cases by ELISA. Crude antigen of T. muris gave better results thanthat of T. spiralis. Immunoblotting can be used for diagnosis of capillariasis by using crude antigen of T.muris.

English

Lymphatic filariasis, commonly termed as elephantiasis has been recorded since ancient Indian, Chinese, Egyptian and Persian writings.1 It is caused by a slender thread like nematode called Wuchereria bancrofti belonging to super family filaroidea.2 It exists in two morphological forms: Adult form and larvae. The adult form has a predilection for intrascrotal lymphatic vessels in hosts; thus hydrocele is one of the most common manifestations of bancroftian filariasis.3 Individuals having circulating microfilaria are outwardly healthy but have the ability to transmit infection to others through mosquito bites (Culex quinquefasciatus) 1 In developing countries like India, occurrence of genital filarial hydrocele is more common in people living in bancroftian endemic areas or in people who were previously operated for eversion of sac. We hereby report a case of hydrocele fluid with microfilaria- an incidental finding on microscopy. Wuchereria bancroti a filarial nematode inhabiting the lymphatics and transmitted through vector - culex mosquito. We hereby present a case of recurrence of genital filariasis. The patient presented with swelling of the left scrotal region with pain of dragging type. On evaluation, eosinophilia was not seen. He had a history of right hydrocele 28 years back. The aspirated hydrocele on direct microscopic examination showed presence of actively motile microfilaria which was confirmed on Giemsa staining. Recurrence is more common in bancroftian filarial endemic areas.

Individual Efficacy and Community Impact of Ivermectin, Diethylcarbamazine, and Albendazole Mass Drug Administration for Lymphatic Filariasis Control in Fiji: A Cluster Randomized Trial.

Bancroftian filariasis remains endemic in Fiji despite >10 years of mass drug administration (MDA) using diethylcarbamazine and albendazole (DA). The addition of ivermectin to this combination (IDA) has improved efficacy of microfilarial clearance at 12 months in individually randomized trials in nocturnal transmission settings, but impact in a setting of diurnally subperiodic filarial transmission has not been evaluated. This cluster randomized study compared the individual efficacy and community impact of IDA vs DA as MDA for lymphatic filariasis in 35 villages on 2 islands of Fiji. Participants were tested at enrollment for circulating filarial antigen and, if positive, for microfilariae. Weight-dosed treatment was offered according to village randomization. Communities were visited at 12 months and retested for lymphatic filariasis. Infected individuals from Rotuma were retested at 24 months. A total of 3816 participants were enrolled and 3616 were treated. At 12 months, microfilariae clearance was achieved in 72 of 111 participants detected with infection at baseline, with no difference in efficacy between treatment groups: DA, 69.2% (95% confidence interval [CI], 57.2%-79.1%) vs IDA, 62.5% (95% CI, 43.6%-78.2%); risk difference, 11.3 % (95% CI, -10% to 32.7%); P = .30. There was no difference between treatment groups in community prevalence of microfilariae at 12 months or individual clearance at 24 months. We found no difference between IDA and DA in individual clearance or community prevalence of lymphatic filariasis at 12 months, and no improved efficacy following a second annual round of IDA. Possible explanations for the apparent lack of benefit of IDA compared to DA include drug and parasite factors affecting clearance, and higher than expected reinfection rates. Clinical Trials Registration: NCT03177993 and Australian New Zealand Clinical Trial Registry: N12617000738325.

Scrotal Filariasis, the Importance of Filarial Dance Sign in Scrotal Ultrasound

Objective: To study the value of the filarial dance sign as a diagnostic sign in scrotal ultrasonography in human Bancroftian filariasis. Methods: We studied 3 patients with scrotal pain, with high-resolution ultrasonography (HRUS) of scrotum including power and color Doppler study. All patients underwent peripheral blood smear. Results: Multiple cystic spaces containing motile echogenic twisted tubular structures were observed in the 3 patients. Peripheral blood smears in the 3 patients confirmed the presence of microfilaria. Mild hydrocele in two patients. Conclusions: HRUS is a useful technique for diagnosing scrotal filariasis in symptomatic patients and is very useful in the follow-up treatment.

Field Comparison between Blood Immunochromatographic Card Test and Microscopy in Diagnosis of Bancroftian Filariasis in Northern Taraba State, Nigeria

Field Comparison between Blood Immunochromatographic Card Test and Microscopy in Diagnosis of Bancroftian Filariasis in Northern Taraba State, Nigeria

A single PCR assay to detect and differentiate Wuchereria bancrofti and Brugia malayi in expatriates and locals with filariasis in Sri Lanka

Background: Lymphatic filariasis is mainly caused by the filarial nematodes Wuchereria bancrofti and Brugia malayi. Sri Lanka was once endemic for bancroftian filariasis, but the disease is nearly forgotten now. However, anti-filariasis Campaign (AFC), Sri Lanka reveals that, bancroftian filariasis still occurs in some selected inlands while there are recent reports of Brugian filariasis. Recently, there has been a large influx of migrant workers from countries endemic for filariasis to Sri Lanka. Therefore, objective of this study was to detect and differentiate W. bancrofti and B. malayi in both expatriates and local filariasis patients by a single PCR. Methods & Materials: Night blood samples received from AFC, Sri Lanka was used for the study after getting approval from relevant authorities. Those blood samples had been collected from migrant workers and local patients who had lymphatic filariasis. DNA was extracted from blood using FavorPrep Blood Genomic DNA Extraction Mini Kit. PCR was performed on extracted DNA using Hha1 and Ssp I primers in single PCR and PCR amplicons were detected by gel electrophoresis. Results of PCR assay were compared with microscopic examination of thick blood films. Results: PCR amplicons were 188 bp and 322 bp for W. bancrofti and B. malayi respectively. The primer specificity was determined against non-filarial parasites and results showed no PCR amplification indicating PCR assay is highly species specific. The minimum number of parasite detectable in blood was less than 12 mf. Results PCR was in line with results of microscopic examination in 90%. Conclusion: This PCR assay found to be highly specific and sensitive in detecting W. bancrofti and B. malayi in expatriate and local filariasis patients and efficient than microscopy. Further this assay can be used for simultaneous detection of both the parasites as a diagnostic tool for the detection of filariasis in population or screening of migrant workers and for rapid surveillance in endemic areas.

Elimination of Lymphatic Filariasis in Shandong Province, China, 1957-2015.

China used to be one of the most heavily endemic countries for lymphatic filariasis (LF) in the world. Bancroftian filariasis, which is caused by the filarioid nematode Wuchereria bancrofti, is the only filariasis in Shandong Province. A total of 864 endemic counties (cities) in 16 provinces/autonomous regions/municipalities with a total population of 330 million people were at risk of infection. Shandong Province was a highly LF-endemic area in the 1950s, the epidemiological investigation of LF conducted in 1957 indicated that the disease was endemic in 74 counties and the highest microfilaria rate was up to 26%. There were ∼5 million people in the province infected with LF; among which almost 2.5 million people had lymphedema, elephantiasis, or hydrocele. Through vigorous prevention and scientific research, Shandong Province was the first to propose to treat LF with fortified diethylcarbamazine salt, more than 25 million people in the province had taken the salt, and more than 3 million people took intermittent medication. After more than 50 years of unremitting efforts, Shandong Province basically eliminated LF in 1983 and became the first province in China that successfully eradicated LF in 2004, which has played an important role in accelerating the elimination of filariasis in the country in 2007 and has made tremendous contributions to social and economic development of China. Since 1980, Shandong Province has carried out extensive international cooperation as the WHO Collaborating Center for Lymphatic Filariasis. This article is intended to share the experience in eliminating LF to other parts of the world and improve public health capacity in regions such as Africa and Oceania where the disease is still endemic for interest.

Molecular xenomonitoring of diurnally subperiodic Wuchereria bancrofti infection in Aedes (Downsiomyia) niveus (Ludlow, 1903) after nine rounds of Mass Drug Administration in Nancowry Islands, Andaman and Nicobar Islands, India.

A group of four human inhabited Nancowry Islands in Nicobar district in the Andaman and Nicobar Islands, India having a population of 7674 is the lone focus of diurnally sub-periodic Wuchereria bancrofti (DspWB) that is transmitted by Aedes niveus (Ludlow). Microfilaria (Mf) prevalence was above 1% even after nine rounds of Mass Drug Administration (MDA) with DEC and albendazole. Molecular xenomonitoring (MX) was conducted to identify appropriate vector sampling method and assess the impact. BioGents Sentinel traps, gravid traps and human baited double bed nettraps were used in three locations in each village to collect Aedes niveus female mosquitoes. Subsequently daytime man landing collections (MLC) were carried out in all the 25 villages in the islands. Collections were compared in terms of the number of vector mosquitoes captured per trap collection. Females of Ae. niveus were pooled, dried and processed for detecting filarial parasite DNA using RT-PCR assay. Vector infection rate was estimated using PoolScreen software. Only 393 female mosquitoes including 44 Ae. niveus (11.2%) were collected from 459 trap collections using three trapping devices. From 151 MLCs, 2170 Ae. niveus female mosquitoes were collected. The average prevalence of W. bancrofti DNA was 0.43%. Estimated upper 95% CI exceeded the provisional prevalence threshold of 0.1% in all the villages, indicating continued transmission as observed in Mf survey. MLCs could be the choice, for now, to sample Ae. niveus mosquitoes. The PCR assay used in MX for nocturnally periodic bancroftian filariasis could be adopted for DspWB. The vector-parasite MX, can be used to evaluate interventions in this area after further standardization of the protocol.

A Study on Serological Reactivity Profile of Different Antigen Preparations with Bancroftian filariasis Human Infection Sera

Lymphatic Filariasis (LF) is one of the incapacitating and mosquito-borne sicknesses that on progression may prompt a few recognizable types of clutters like extreme lymphedema, hydrocele, and elephantiasis. Antigenic preparations of B. malayi adult (BmA), S. cervi adult parasites and microfilariae (mf) total parasite extract were used to analyze the serological reactivity profile with human infectious sera collected from endemic areas of Bancroftian filariasis by performing Western blot and ELISA analysis. Sera from healthy human subjects were also included in the study to determine the variation incurred in the reactivity due to the filariasis infection. Gelelectrophoresis analysis of the crude-extract of BmA revealed seven protein bands while more than ten bands were recognized in S. cervi. our results represent a clear variation in protein patterns among the crude-antigens. ELISA results showed highest prevalence of IgG, IgM and IgG4 antibodies against all antigen preparations when recorded among microfilaraemic chronic infected patients. In both the antigenic preparations, the positive reactions were in the order of microfilaraemic>endemic normal>chronic>acute>nonendemic normal subjects. All sera of Mf+ patients were uniformly positive, while sera of both chronic and endemic normal subjects showed less reactivity. In the present study, we endeavoured to establish the extent of cross-reactivity of antigens derived from animal filarial parasites such as B. malayi and S. cervi with W. bancrofti filariasis sera of human patients. Besides, we further analyzed antibody-isotype profile of IgG, IgG4 and IgM in various human infection sera of bancroftian filarial subjects reactive to heterologous parasite antigens derived from adult worms of S. cervi from bovine and B. malayi from bovine and jirds.