Histological validation of in-vivo larvicidal efficacy of marine Streptomyces sp. RD06 secondary metabolites against filariasis causing Culex quinquefasciatus and statistical media optimization for larvicidal derivatives production.

Mosquito-borne disease pandemics, such as the Zika virus and chikungunya, have escalated cognizance of how critical it is to implement proficient mosquito vector control measures. The prevention of Culicidae is becoming more difficult these days because of the expeditious imminence of synthetic pesticide resistance and the universal expansion of tremendously invasive mosquito vectors. The present study highlights the insecticidal and larvicidal efficacy of the prospective novel actinobacterium derived from the marine Streptomyces sp. RD06 secondary metabolites against Culex quinquefasciatus mosquito. The pupicidal activity of Streptomyces sp. RD06 showed LC50=199.22 ± 11.54 and LC90= 591.84 ± 55.41 against the pupa. The purified bioactive metabolites 1, 2-Benzenedicarboxylic acid, diheptyl ester from Streptomyces sp. RD06 exhibited an LC50 value of 154.13 ± 10.50 and an LC90 value of 642.84 ± 74.61 tested against Cx. quinquefasciatus larvae. The Streptomyces sp. RD06 secondary metabolites exhibited 100 % non-hatchability at 62.5 ppm, and 82 % of hatchability was observed at 250 ppm. In addition, media optimization showed that the highest biomass production was attained at a temperature of 41.44 °C, pH 9.23, nitrogen source 11.43 mg/ml, and carbon source 150 mg/ml. Compared to control larvae, the histology and confocal microscopy results showed destruction to the anal gill, lumen content, and epithelial layer residues in the treated larvae. Utilizing an eco-friendly method, these alternative inventive insecticidal derivatives from Streptomyces sp. RD06 eradicates Culex quinquefasciatus. This study highlights the promising potential of these Streptomyces sp. RD06 secondary metabolites to develop affordable and efficacious mosquito larvicides to replace synthetic insecticides in the future.

Highlighting the Relevance of CD8+ T Cells in Filarial Infections.

The T cell immune responses in filarial infections are primarily mediated by CD4+ T cells and type 2-associated cytokines. Emerging evidence indicates that CD8+ T cell responses are important for anti-filarial immunity, however, could be suppressed in co-infections. This review summarizes what we know so far about the activities of CD8+ T cell responses in filarial infections, co-infections, and the associations with the development of filarial pathologies.

Filariasis and transfusion-associated risk: a literature review.

BACKGROUND AND OBJECTIVES: Filariae are parasitic worms that include the pathogens Loa loa, Onchocerca volvulus, Wuchereria bancrofti, Brugia spp. and Mansonella spp. which are endemic in parts of Africa, Asia, Asia-Pacific, South and Central America. Filariae have a wide clinical spectrum spanning asymptomatic infection to chronic debilitating disease including blindness and lymphedema. Despite successful eradication programmes, filarial infections remain an important -albeit neglected - source of morbidity. We sought to characterize the risk of transfusion transmission of microfilaria with a view to guide mitigation practices in both endemic and non-endemic countries. MATERIALS AND METHODS: A scoping review of scientific publications as well as grey literature was carried out by a group of domain experts in microbiology, transfusion medicine and infectious diseases, representing the parasite subgroup of the International Society of Blood Transfusion. RESULTS: Cases of transfusion-transmitted filariasis are rare and confined to case reports of variable quality. Transfusion-associated adverse events related to microfilariae are confined to isolated reports of transfusion reactions. Serious outcomes have not been reported. No known strategies have been implemented, specifically, to mitigate transfusion-transmitted filariasis yet routine blood donor screening for other transfusion-transmissible infections (e.g. hepatitis B, malaria) may indirectly defer donors with microfilaremia in endemic areas. CONCLUSION: Rare examples of transfusion-transmitted filariasis, without serious clinical effect, suggest that filariasis poses low transfusion risk. Dedicated mitigation strategies against filarial transfusion transmission are not recommended. Given endemicity in low-resource regions, priority should be on the control of filariasis with public health measures.

The immune response of inbred laboratory mice to Litomosoides sigmodontis: A route to discovery in myeloid cell biology.

Litomosoides sigmodontis is the only filarial nematode where the full life cycle, from larval delivery to the skin through to circulating microfilaria, can be completed in immunocompetent laboratory mice. It is thus an invaluable tool for the study of filariasis. It has been used for the study of novel anti-helminthic therapeutics, the development of vaccines against filariasis, the development of immunomodulatory drugs for the treatment of inflammatory disease and the study of basic immune responses to filarial nematodes. This review will focus on the latter and aims to summarize how the L sigmodontis model has advanced our basic understanding of immune responses to helminths, led to major discoveries in macrophage biology and provided new insights into the immunological functions of the pleural cavity. Finally, and most importantly L sigmodontis represents a suitable platform to study how host genotype affects immune responses, with the potential for further discovery in myeloid cell biology and beyond.

Effect of tiliamosine, a bis, benzylisoquinoline alkaloid isolated from Tiliacora acuminata (Lam.) Hook. f. & Thom on the immature stages of filarial mosquito Culex quinquefasciatus say (Diptera: Culicidae).

The present study was aimed to check the mosquitocidal activity of tiliamosine isolated from Tiliacora acuminata (Lam.) Hook. f. & Thom against immature stages of Culex quinquefasciatus. Eggs and larvae of Cx. quinquefasciatus were exposed to different concentrations of tiliamosine - 0.5, 1.0, 1.5 and 2.0 ppm - prepared using DMSO. The compound tiliamosine showed good larvicidal activity with LC50 and LC90 values of 1.13 and 2.85 ppm respectively, against third-instar larvae of Cx. quinquefasciatus at 24 h. In control, the larvae exhibited normal movement. Tiliamosine exhibited 91% ovicidal activity at 2.0 ppm concentration after 120 h post-treatment. Lowest concentration of tiliamosine (0.5 ppm) showed 19% egg mortality. Histopathology study of the compound-treated larvae showed serious damage on the larval midgut cells. The treated larvae showed restless movement which was different from that of the control larvae. The larvae exhibited malformation in development. The compound tiliamosine was harmless to non-target organisms P. reticulata and Dragon fly nymph at tested concentrations. The compound was highly active and inhibited AChE in a concentration-dependent manner. Computational analysis of the tiliamosine had strong interaction with AChE1 of Cx. quinquefasciatus. This report clearly suggests that the isolated compound can be used as an insecticide to control mosquito population and thus prevent the spread of vector-borne diseases.

Efficacy of subcutaneous doses and a new oral amorphous solid dispersion formulation of flubendazole on male jirds (Meriones unguiculatus) infected with the filarial nematode Brugia pahangi.

River blindness and lymphatic filariasis are two filarial diseases that globally affect millions of people mostly in impoverished countries. Current mass drug administration programs rely on drugs that primarily target the microfilariae, which are released from adult female worms. The female worms can live for several years, releasing millions of microfilariae throughout the course of infection. Thus, to stop transmission of infection and shorten the time to elimination of these diseases, a safe and effective drug that kills the adult stage is needed. The benzimidazole anthelmintic flubendazole (FBZ) is 100% efficacious as a macrofilaricide in experimental filarial rodent models but it must be administered subcutaneously (SC) due to its low oral bioavailability. Studies were undertaken to assess the efficacy of a new oral amorphous solid dispersion (ASD) formulation of FBZ on Brugia pahangi infected jirds (Meriones unguiculatus) and compare it to a single or multiple doses of FBZ given subcutaneously. Results showed that worm burden was not significantly decreased in animals given oral doses of ASD FBZ (0.2-15 mg/kg). Regardless, doses as low as 1.5 mg/kg caused extensive ultrastructural damage to developing embryos and microfilariae (mf). SC injections of FBZ in suspension (10 mg/kg) given for 5 days however, eliminated all worms in all animals, and a single SC injection reduced worm burden by 63% compared to the control group. In summary, oral doses of ASD formulated FBZ did not significantly reduce total worm burden but longer treatments, extended takedown times or a second dosing regimen, may decrease female fecundity and the number of mf shed by female worms.

[Current status of chronic filariasis in Jiangsu Province].

OBJECTIVE: To understand the current status of chronic filariasis patients in Jiangsu Province so as to provide basic data for following-up care for them. METHODS: The patients were followed up one by one according to history archives between June and July, 2018, and the clue investigation was also conducted. The base data of the patients was collected through a face-to-face questionnaire survey and analyzed. RESULTS: There were still 3 160 chronic filariasis patients in Jiangsu Province. Among them, the male accounted for 40.0%, and 91.8% of the patients were older adults aged 60 years or above. From the aspect of regional distribution, Suqian (24.2%), Huai'an (19.5%), Suzhou (17.3%), Xuzhou (11.2%), and Yancheng (9.8%) were the five top high prefectures. The patients with simple lymphatic inflammation or lymphadenitis, simple lymphedema or elephantiasis, simple chyluria, simple hydrocele of tunica vaginalis, and two symptoms or more accounted for 2.7%, 37.1%, 11.2%, 0.9%, and 48.1%, respectively. For the patients with lymphedema or elephantiasis, 97.8% of edema was seen in the lower limbs, and more than 90% of the edema stages were I-III. The number of current caring sites was 220, covering 2 091 patients. The average number of times of caring activities in this year was 3.2. The average cumulative time of caring activities among all the sites was 11.3 years. CONCLUSIONS: The number of chronic filariasis patients has been dramatically decreased, most of the patients are old and have long disease durations. The caring sites have not covered all the patients. In order to release the symptoms and improve the life quality of the patients, all the patients should be taken care of in Jiangsu Province.

Ligand binding properties of two Brugia malayi fatty acid and retinol (FAR) binding proteins and their vaccine efficacies against challenge infection in gerbils.

Parasitic nematodes produce an unusual class of fatty acid and retinol (FAR)-binding proteins that may scavenge host fatty acids and retinoids. Two FARs from Brugia malayi (Bm-FAR-1 and Bm-FAR-2) were expressed as recombinant proteins, and their ligand binding, structural characteristics, and immunogenicities examined. Circular dichroism showed that rBm-FAR-1 and rBm-FAR-2 are similarly rich in α-helix structure. Unexpectedly, however, their lipid binding activities were found to be readily differentiated. Both FARs bound retinol and cis-parinaric acid similarly, but, while rBm-FAR-1 induced a dramatic increase in fluorescence emission and blue shift in peak emission by the fluorophore-tagged fatty acid (dansyl-undecanoic acid), rBm-FAR-2 did not. Recombinant forms of the related proteins from Onchocerca volvulus, rOv-FAR-1 and rOv-FAR-2, were found to be similarly distinguishable. This is the first FAR-2 protein from parasitic nematodes that is being characterized. The relative protein abundance of Bm-FAR-1 was higher than Bm-FAR-2 in the lysates of different developmental stages of B. malayi. Both FAR proteins were targets of strong IgG1, IgG3 and IgE antibody in infected individuals and individuals who were classified as endemic normal or putatively immune. In a B. malayi infection model in gerbils, immunization with rBm-FAR-1 and rBm-FAR-2 formulated in a water-in-oil-emulsion (®Montanide-720) or alum elicited high titers of antigen-specific IgG, but only gerbils immunized with rBm-FAR-1 formulated with the former produced a statistically significant reduction in adult worms (68%) following challenge with B. malayi infective larvae. These results suggest that FAR proteins may play important roles in the survival of filarial nematodes in the host, and represent potential candidates for vaccine development against lymphatic filariasis and related filarial infections.

Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) extracts exhibit strong larvicidal activity against mosquito vectors of malaria, dengue fever, and filariasis.

Vector-borne diseases transmitted by mosquitoes cause globally important diseases such as malaria, dengue fever, and filariasis. The incidence of these diseases can be reduced through mosquito control programs but these control programs currently rely on synthetic insecticides that can impact the environment, and has selected widespread mosquito resistance. Environment friendly and biodegradable natural insecticides discovered in plants offer an alternative approach to mosquito control. Here, we investigated extracts from root or aerial parts of Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) against the early 4th instar larvae of Anopheles stephensi (malaria vector), Aedes aegypti (dengue fever vector), and Culex quinquefasciatus (filariasis vector). The root and aerial parts extracts of A. absinthium and C. intybus at 200, 100, 50, 25 and 12.5 ppm caused significant mortality of the tested mosquito species. Root extracts exhibited higher larvicidal activity that aerial part extracts. The highest larvicidal activity was recorded in methanol extract of roots of C. intybus with LC50 = 66.16, 18.88 and LC¬90 = 197.56, 107.16 ppm for An. stephensi; LC50 = 78.51, 40.15 and LC90 = 277.31, 231.28 ppm for Ae. aegypti and LC50 = 103.99, 64.56 and LC¬90 = 314.04, 247.54 ppm for Cx. quinquefasciatus. These results reveal potent mosquito larvicidal activity against vectors of malaria, dengue fever, and filariasis is present in extracts of chicory and wormwood.

Prospects of developing a prophylactic vaccine against human lymphatic filariasis - evaluation of protection in non-human primates.

Lymphatic filariasis (LF) affects 120 million people around the world and another 856 million people are at risk of acquiring the infection. Mass Drug Administration (MDA) spearheaded by the World Health Organization is the only current strategy to control this infection. Recent reports suggest that despite several rounds of MDA, elimination has not been achieved and there is a need for more stringent control strategies for control of LF. An effective prophylactic vaccine combined with MDA has significant potential. Initial trials using a prophylactic trivalent recombinant Brugia malayi heat shock protein 12.6, abundant larval transcript -2 and tetraspanin large extra-cellular loop (rBmHAT) vaccine developed in our laboratory conferred only 35% protection in macaques. Therefore, the focus of the present study was to improve the current vaccine formulation to obtain better protection in non-human primates. We made two modifications to the current formulation: (i) the addition of another antigen, thioredoxin peroxidase-2 (TPX-2) to make it a tetravalent vaccine (rBmHAXT) and (ii) the inclusion of an adjuvant; AL019 (alum plus glucopyranosyl lipid adjuvant-stable emulsion) that is known to promote a balanced Th1/Th2 response. A double-blinded vaccination trial was performed with 40 macaques that were divided into three treatment groups and one control group (n = 10/group). Vaccinated animals received 4 immunisations at 1 month intervals with 150 µg/ml of rBmHAT plus alum, rBmHAT plus AL019 or rBmHAXT plus AL019. Control animals received AL019 only. All vaccinated macaques developed significant (P ≤ 0.003) titers of antigen-specific IgG antibodies (1:20,000) compared with the controls. One month after the last dose, all macaques were challenged s.c. with 130-180 B. malayi L3s. Our results showed that seven out of 10 (70%) of macaques given the improved rBmHAXT vaccine did not develop the infection compared with AL019 controls, of which seven out of 10 macaques developed the infection. Titers of antigen-specific IgG1 and IgG2 antibodies were significantly (P ≤ 0.01) higher in vaccinated animals and there was an increase in the percentage of IL-4 and IFN-γ secreting antigen-responding memory T cells. These studies demonstrated that the improved formulation (rBmHAXT plus AL019) is a promising vaccine candidate against human lymphatic filariasis.

[Challenge and strategy of prevention and control of important parasitic diseases under the Belt and Road Initiative].

China was once a country with the heaviest burden of parasitic diseases. Under the leadership of the Communist Party and national authority, after more than 60 years' efforts of prevention and control, the remarkable results have been achieved in China. However, affected by the social and economic development and environmental changes, the prevention and control of parasitic diseases, especially imported parasitic diseases, are facing new challenges, and the parasitic diseases, such as malaria, schistosomiasis, leishmaniasis, filariasis and trypanosomiasis, appear increasingly. With the development of the Belt and Road Initiative, the transmission risks of these diseases are more increased. The purpose of this paper is to describe the experience and results of parasitic disease prevention and control in China, understand the present parasitic disease epidemic situation of the Belt and Road Initiative related countries, analyze the transmission risks of important parasitic diseases, and present some relevant suggestions, so as to provide the evidence for the health administrative department formulating the prevention and control strategies of such parasitic diseases timely and effectively.

Toxicity studies of nanofabricated palladium against filariasis and malaria vectors.

The present study was carried out to establish the biofabrication of palladium nanoparticles (PdNPs) using the plant leaf extract of Tinospora cordifolia Miers and its toxicity studies on the larvae of filariasis vector, Culex quinquefasciatus Say and malaria vector, Anopheles subpictus Grassi. The biofabricated PdNPs were characterized by using UV-visible spectrum, FTIR, XRD, FESEM, EDX and HRTEM. HRTEM confirmed the PdNPs were slightly agglomerated and spherical in shape and the average size was 16 nm. Gas chromatography and mass spectrometry analysis result revealed that the major constituent present in the T. cordifolia leaf extract is 2,4-di-tert-butylphenol (31.79%) whereas the minor compounds are 1-hexadecanol (7.97%), 1-octadecanol (7.70%), 1-eicosanol (6.85%), behenic alcohol (5.36%), 1-tetradecene (6.22%), cyclotetradecane (6.23%), 1-hexadecene (7.97%), 1-octadecene (7.70%), 1-eicosene (6.85%), and 1-docosene (5.36%). T. cordifolia leaf extract exhibited the larvicidal activity against the fourth instar larvae of C. quinquefasciatus and A. subpictus with the values of LC50 = 59.857 and 54.536 mg/L; LC90 = 113.445 and 108.940 mg/L, respectively. The highest toxicity was observed in the biofabricated PdNPs against the fourth instar larvae of C. quinquefasciatus and A. subpictus with the values of LC50 = 6.090 and 6.454 mg/L; LC90 = 13.689 and 13.849 mg/L, respectively. Concerning non-target effects, Poecilia reticulata were exposed to PdNPs for 24 h and did not exhibit any noticeable toxicity. Overall, our findings strongly suggest that PdNPs is a perfect ecological and inexpensive approach for the control of filariasis and malaria vectors.

Mining Filarial Genomes for Diagnostic and Therapeutic Targets.

Filarial infections of humans cause some of the most important neglected tropical diseases. The global efforts for eliminating filarial infections by mass drug administration programs may require additional tools (safe macrofilaricidal drugs, vaccines, and diagnostic biomarkers). The accurate and sensitive detection of viable parasites is essential for diagnosis and for surveillance programs. Current community-wide treatment modalities do not kill the adult filarial worms effectively; hence, there is a need to identify and develop safe macrofilaricidal drugs. High-throughput sequencing, mass spectroscopy methods and advances in computational biology have greatly accelerated the discovery process. Here, we describe post-genomic developments toward the identification of diagnostic biomarkers and drug targets for the filarial infection of humans.

Potential Role for Flubendazole in Limiting Filariasis Transmission: Observations of Microfilarial Sensitivity.

Flubendazole (FLBZ) is a potent and efficacious macrofilaricide after parenteral administration. Studies in animal models and one trial in patients infected with Onchocerca volvulus revealed that FLBZ elicits minimal effects on microfilariae (mf). Severe complications after ivermectin (IVM) treatment of patients with high Loa loa microfilaraemia are of great concern. We examined the potential of FLBZ to rapidly kill L. loa mf, the phenomenon proposed to underlie the complications. Mf of L. loa were exposed to FLBZ, its reduced metabolite, albendazole, or IVM in vitro. Viability of L. loa mf was unaffected by FLBZ (10 µM, 72 hours); similar results were obtained with mf of Brugia malayi. We also measured the effects of FLBZ on transmission of mf. Aedes aegypti were fed FLBZ-exposed B. malayi mf and dissected 24 hours or 14 days postfeeding to count mf that crossed the midgut and developed to infective L3. FLBZ impaired the ability of mf to cross the midgut, regardless of duration of exposure (≥ 2 hours). FLBZ also prevented the development of mf to L3s, irrespective of duration of exposure or concentration. FLBZ is not microfilaricidal under these conditions; however, it blocks transmission. These results support the possibility that FLBZ may be a useful macrofilaricide in loiasis regions and may limit transmission from treated individuals.

Ivermectin induced Steven-Johnsons syndrome: case report.

BACKGROUND: Stevens-Johnson syndrome is one of the manifestations of mucocutaneous adverse drug reactions. Although antimicrobials are responsible for greater than 50% of these adverse drug reactions, there is no documented case implicating ivermectin as the culprit. A 38 year old adult Cameroonian male presented to our health facility with facial rash, painful oral sores, black eschars on lips and red tearing eyes 3 days following ingestion of ivermectin received during a nationwide anti-filarial campaign. He had no known chronic illness, no known allergies and was not on any medications prior to the campaign. Physical examination revealed discharging erythematous eyes, crusted and blister-like lesions with cracks on his lips and oral mucosa. His laboratory tests were unremarkable but for a positive Human Immunodeficiency Virus (HIV) test. A diagnosis of Ivermectin induced Stevens-Johnson syndrome in a newly diagnosed HIV patient was made. The patient was managed with supportive therapy and the evolution thereafter was favourable. CONCLUSION: Stevens-Johnson syndrome is a potential side effect of ivermectin and susceptibility to this adverse effect may be increased in HIV infection.

Travel-Related Parasitic Infections in Travellers to Southeast Asia and Western Pacific Countries.

In the last decades, there has been a significant increase in international human mobility with increase in the prosperity, travel possibilities, and number of refugees. In the first half of 2016, the Asian continent showed the fastest growth in the number of tourists. Such increase is seen due to the interest in Asian history, culture, and cuisine. In the globalizing world, human mobility causes changes in the epidemiology of diseases and the spread of various infections across continents. Parasitic infections that may pose a risk for travellers to the Asia-Pacific are malaria, leishmaniasis, filariasis, foodborne trematode infections, schistosomiasis, soil-transmitted infections, and tourist diarrhea. Consulting a travel medical expert and using health services such as pre-travel vaccination and chemoprophylaxis will reduce the risk of infectious diseases among travelers.