Filarial Parasite Research Articles

An Initiative the Used of Sterile Insect Technique (SIT) To Control Filariasis in Indonesia

Filariasis as part of the neglected tropical disease is one of the health problems in the world. Filariasis divided into onchocerciasis (river blindness) and lymphatic filariasis or elephantiasis. This disease caused by filarial nematode parasites Onchocerca volvulus, Wuchereria bancrofti, Brugia malayi and Brugia timori. Filariasis transmitted by several mosquito genera as the vector. Indonesia as endemic filariasis, agreed on plays a role on World Health Organization (WHO) global filariasis elimination in 2020. Sterile insect technique (SIT) is a potential method which can be applied to filariasis elimination program by controlling the mosquito population. Basic principles of SIT involve mass rearing of species target, sterilization process using gamma-rays and releasing sterile male insect into a target area. SIT combined with other methods under one management as Area-wide Integrated Pest Management (AW-IPM) to increasing effectiveness and successful filariasis elimination program in Indonesia. Filariasis elimination program in Indonesia has several challenges and needed public participation to achieve program goals.

An ARV1 homologue from a filarial nematode is functional in yeast.

The transmembrane protein, ARV1, plays a key role in intracellular sterol homeostasis by controlling sterol distribution and cellular uptake. To date, only the ARV1s from yeast and humans have been characterized to some extent. In this study, the ARV1 of an animal filarial parasite, Setaria digitata (SdARV1), was characterized; its cDNA was 761 bp and encoded a protein of 217 amino acids, with a predicted molecular weight of 25 kDa, containing a highly conserved ARV1 homology domain and three transmembrane domains in the bioinformatic analyses. Information required to cluster members belonging to a particular taxon has been revealed in phylogenetic analyses of ARV1 sequences derived from different organisms. Reverse transcription-polymerase chain reaction (RT-PCR) analyses indicated that SdARV1 was expressed in different developmental stages - microfilariae and adult male and female worms. Experiments carried out with a single copy of the SdARV1 under the control of the PMA-1 promoter in a temperature-sensitive Saccharomyces cerevisiae mutant strain indicated full complementation of the mutant phenotype, with growth at a non-permissive temperature (37°C). Microscopic observations of cellular morphology with Gram staining revealed alteration of the shape from shrunken to oval, in mutant and complemented strains, respectively. Assessment of free sterol levels extracted from mutant yeast and complemented strains indicated that the level of sterol was significantly higher in the former compared to the latter, which had sterol levels similar to those of the wild type. Thus, the results of the current study suggest that SdARV1 is ubiquitously expressed in different developmental stages of S. digitata, and that it is a true functional homologue of mammalian and yeast ARV1s, which have crucial phylogenetic information that follows classical evolutionary trends. Finally, this is the first study to report the biological function of nematode ARV1.

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum, Brugia malayi or Trypanosoma brucei brucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces.

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum, Brugia malayi or Trypanosoma bruceibrucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

Exploration of antifilarial activity of gold nanoparticle against human and bovine filarial parasites: A nanomedicinal mechanistic approach

The present work seeks to explore the antifilarial activity of biopolymer functionalized gold nanoparticles (AuNPs) against human filarial parasite (Wuchereria bancrofti) through Nrf2 signaling for the first time. A natural polymer, chitosan is used along with Terminalia chebula extract to synthesize AuNPs following the principles of green chemistry. The probable mode of action of AuNPs as filaricidal agent has been investigated in detail using model filarial parasite, Setaria cervi (bovine parasite). Biopolymers inspired AuNPs exhibit superior antifilarial activity against both human and bovine filarial parasites, and are able to induce oxidative stress and apoptotic cell death in filarial parasites mediated through mitochondria. AuNPs also alter the Nrf2 signaling. In addition, the synthesized nanomaterials appear to be nontoxic to mammalian system. Thus the present mechanistic study, targeting human filarial parasites, has the potential to increase the therapeutic prospects of AuNPs to control lymphatic filariasis in the upcoming days.

Thioredoxin reductase from the bovine filarial parasite Setaria cervi: Studies on its localization and optimization of the extraction

Exploration of novel drug targets has been the major thrust area in filarial research. In this regard, identification and characterization of oxidative enzymes that play pivotal role in the survival of filarial parasite inside host are of immense importance. In this study, we are reporting the presence of an important redox regulatory enzyme, thioredoxin reductase (TrxR) in the bovine filarial parasite Setaria cervi. TrxR was found to be exists throughout the developmental stages viz. oocyte, microfilaria and adult of the parasite. Since further studies on this enzyme require adequate quantity, influential extraction parameters were optimized statistically using response surface methodology (RSM) employing a seven factors based Box-Behnken design matrix. ANOVA analysis revealed the relative importance of each parameter and a regression equation was eventually developed that could predict the specific activity (SA) of TrxR. Finally the optimized extraction conditions predicted by RSM was 6.1ml of 61.86mM buffer, pH 6.0, with extraction temperature 39.96°C for 180min in addition to 450rpm agitation and 20μl/ml of protease inhibitor. Therefore this study is going to be the maiden report depicting the identity of TrxR in filarial parasite and the optimized extraction conditions for its isolation with better kinetic efficiency.

Immune response in Mansonella ozzardi infection modulated by IL-6/IL-10 axis in Amazon region of Brazil

Mansonellosis is an endemic disease in the South and Central America. In Brazil, one of the etiological agents is Mansonella ozzardi. This filarial infection is yet poorly understood, with a controversial morbity, presenting since a oligosymptoms, malaria-like signs or without complaint in humans. The knowledge of the human immune response to microfilariae infection is limited mainly by different evolutionary cycles of the parasite in the host. In addition, the prevalence of this filarial parasite infection is high in several regions of Amazonas State. A cross-sectional study was conducted in an endemic area for microfilariae of M. ozzardi (MF) infection in the Amazonas State, Brazil. Proinflammatory and regulatory cytokines (IL-2, IL-4, IL-6, IL-10, TNF, IFN-gamma, and IL-17A) were measured in cryopreserved serum using the Cytometric Bead Array techniques (CBA) in 54 patients diagnosed with M. ozzardi infection and 55 individuals without the infection were included in the study (Controls). The IL-4, IL-6 and IL-10 level increased in infected patients with MF infection, while IL-17A increased in control only. When we compared controls to patients with high or low parasite load, the increased level of IL-6 and IL-10 were maintained. IL-6 contributes to the proinflammatory activity and IL-10 modulates Th1, Th2 and Th17 immune response. Furthermore, IL-4 was detected as a marker in the MF infection and MF patients with low parasite load, indicating the action of the Th2 cell response. The complex network of cytokines acting during M. ozzardi infection depends on a fine balance to determine a host protective effect or filarial persistence. Therefore, these results suggest that the immune response in MF infection is modulated by IL-6/IL-10 axis.

Detection of Cercopithifilaria bainae in western Romania.

Cercopithifilaria species are tick-transmitted filarial parasites of mammals. In Europe, three Cercopithifilaria spp. are known to parasitize dogs, all occurring mainly in the Mediterranean countries. In Romania, Cercopithifilaria bainae has been reported in a single dog in eastern Romania but the occurrence in other parts of the country is not known. To further elucidate the geographic distribution of Cercopithifilaria spp. infection, 544 ticks were collected from dogs in several locations across Romania. The presence of Cercopithifilaria spp. was investigated with real-time PCR. A single Dermacentor reticulatus female tick was found to be infected with Cercopithifilaria bainae. The finding in the present study is geographically separated from the previous finding in Romania by 800km, as well as by the Carpathian mountain range. Hence, C. bainae is more geographically widespread in Romania than previously recognized. However, the single detection does suggest that infection is rather uncommon in Romanian dogs. Nevertheless, further studies on Cercopithifilaria spp. distribution and prevalence are needed.

Recombinant Brugia malayi pepsin inhibitor (rBm33) exploits host signaling events to regulate inflammatory responses associated with lymphatic filarial infections

Prolonged existence of filarial parasites and their molecules within the host modulate the host immune system to instigate their survival and induce inflammatory responses that contribute to disease progression. Recombinant Brugia malayi pepsin inhibitor (rBm33) modulates the host immune responses by skewing towards Th1 responses characterized by secretion of inflammatory molecules such as TNF-α, IL-6, nitric oxide (NO). Here we also specified the molecular signaling events triggered by rBm33 in peripheral blood mononuclear cells (PBMCs) of filarial endemic normals (EN). rBm33 predominantly enhanced the levels of nitric oxide in cultured PBMCs but did not result in oxidative stress to the host cells. Further, rBm33 treatment of human PBMCs resulted in higher GSH/GSSG levels. MYD88 dependent activation was found to be associated with rBm33 specific inflammatory cytokine production. rBm33 triggered intracellular signaling events also involved JNK activation in host PBMCs. In addition, c-Fos and not NF-κB was identified as the transcription factor regulating the expression of inflammatory cytokines in rBm33 stimulated PBMCs. rBm33 marked its role in filarial pathology by altered levels of growth factors but did not have a significant impact on matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs) activity of host PBMCs. Thus, the study outlines the signaling network of rBm33 induced inflammatory responses within the host immune cells.

Studying the Biological Activities and Molecular Docking of Some Novel Benzosultams and Benzosultones

Background: The antibacterial, antifungal and antiparasitic activities of some novel benzosultams and benzosultones in vitro systems are tested. The antibacterial activity has been tested against gram negative bacterium Escherichia coli (E. coli) and gram positive Bacillus subtilis (B. subtilis). The antifungal activity has been tested against an opportunistic pathogenic fungal strain namely Pichia guilliermondii SNP3S. The antiparasitic activity has been tested against filarial parasite Setaria cervi. The active compound 4,7-dimethyl-3,4,7,8-tetrahydro-3λ6-[1,2]thiazino[4,3-f]quinoline-3,3,8-trione was found to exert no toxicity when tested on rat peritoneal macrophage culture. The compounds show moderate to excellent biological activities. The molecular modeling has also been done to compare their inhibitory affinity on human matrix metalloproteinase 9 (MMP-9). Keywords: Antibacterial, antifungal, antiparasitic, benzosultams, benzosultones, molecular modeling, binding affinity.

Imported Infections with Mansonella perstans Nematodes, Italy.

We report 74 patients in Italy infected with Mansonella perstans nematodes, a poorly described filarial parasite. M. perstans nematodes should be included in the differential diagnosis for patients with eosinophilia from disease-endemic countries. Serologic analysis is useful for screening, and testing for microfilaremia in peripheral blood should be performed for parasite-positive patients.

P038 Certain KIR-HLA genotypes suggestive of increased natural killer cell responses are associated with tuberculosis

Aim KIR-HLA variation affects KIR repertoire of NK cell clones, NK cell maturation and thus influence the capability to deliver signals, which may impact NK cell response on infections. Aim of this study is to test if certain KIR-HLA combinations are involved in the risk of developing pulmonary tuberculosis (PTB). Methods We typed 375 PTB patients and 249 healthy controls collected from All India Institute of Medical Sciences, India for KIR and HLA class I genes using Luminex rSSO. All control subjects were positive to the purified protein derivative (PPD), but did not suffer from any known disease and their chest radiograph was normal. Results Telomeric half of KIR complex was associated with PTB. Specifically, an increase of B -haplotype associated KIRs (3DS1, 2DS1) and a decrease of A -haplotype associated KIRs (3DL1, 2DS4) were observed in PTB (Table). Moreover, a substantial increase of 2DL1 + C2 (p = 0.0001) and a decrease of 3DL1 + Bw4 (p = 0.023) were observed in PTB. These results suggest the possibility of generating more activating NK cells by 2DL1 + C2-mediated licensing plus expressing activating KIR 3DS1/2DS1. Conclusions These findings contrast the classical view that the activating KIR mediate spontaneous lysis of infected cells and provide protection against the infection. The NK cells expressing a dominant activating KIR repertoire may presumably trigger a localized hyperresponsiveness leading to the secretion of cytokines and growth factors that likely exacerbating PTB disease. Consistent with this unexpected findings, a detrimental role for NK cells against non-viral pathogens has been suggested from the following studies: (i) depletion of NK cells was reported to increase the ability of the host to control Listeria monocytogenes (J Immunol. 1994, 152:1873); (ii) host NK cells were found to be required for the growth of the human filarial parasite Brugia malayi (J Immunol. 1998, 161:1428). Together, these findings suggest a revision of our opinion of the role of NK cells in non-viral parasitic diseases. Download : Download high-res image (192KB) Download : Download full-size image

Occurrence and diversity of arthropod-transmitted pathogens in red foxes (Vulpes vulpes) in western Austria, and possible vertical (transplacental) transmission of Hepatozoon canis.

Red fox (Vulpes vulpes) is the most abundant wild canid species in Austria, and it is a well-known carrier of many pathogens of medical and veterinary concern. The main aim of the present study was to investigate the occurrence and diversity of protozoan, bacterial and filarial parasites transmitted by blood-feeding arthropods in a red fox population in western Austria. Blood (n = 351) and spleen (n = 506) samples from foxes were examined by PCR and sequencing and the following pathogens were identified: Babesia canis, Babesia cf. microti (syn. Theileria annae), Hepatozoon canis, Anaplasma phagocytophilum, Candidatus Neoehrlichia sp. and Bartonella rochalimae. Blood was shown to be more suitable for detection of Babesia cf. microti, whilst the spleen tissue was better for detection of H. canis than blood. Moreover, extremely low genetic variability of H. canis and its relatively low prevalence rate observed in this study may suggest that the parasite has only recently been introduced in the sampled area. Furthermore, the data presented here demonstrates, for the first time, the possible vertical transmission of H. canis from an infected vixen to the offspring, and this could explain the very high prevalence in areas considered free of its main tick vector(s).

Therapeutic trial of doxycyclin plus ivermectin for the treatment of Brugia malayi naturally infected cats

Lymphatic filariasis (LF) is one of the neglected tropical diseases which causes permanent and long term disability worldwide. LF is caused by filarial nematode parasites, i.e. Wuchereria bancrofti, Brugia malayi, and B. timori. All available antifilarial drugs currently being used have shown a limited adulticidal activity. Discoveries of endosymbiont rickettsia-like bacterium, Wolbachia in filarial nematodes provided a novel approach for antibiotic use in eradication of filarial diseases. The earlier studies revealed the macrofilaricidal efficacy of doxycycline against filarial nematodes. Chemotherapeutic efficiency of doxycycline has been studied against many filarial parasites, but there are still no therapeutic trials of the drug regimens for B. malayi treatment in naturally infected cats. Thus, this study would be the first attempt to study the efficiency of doxycycline (DOXY) alone or in combination with ivermectin (IVM) for treatment of B. malayi in naturally infected cats. A total of 26 B. malayi-infected cats in the endemic areas were recruited and divided into 3 groups, receiving different treatment regimens; a single dose of ivermectin only (IVM), doxycycline only (DOXY) and a combination of ivermectin and doxycycline (DOXY-IVM). The efficacy of each therapatic regimen was evaluated by detecting the presence of microfilaria using parasitological and molecular techniques monthly up to 2 years after starting the treatment. The IVM treated group had a significant rapid reduction of microfilariae in the first month; however, recurrence of microfilaraemia was observed in some cats. By contrast, the DOXY and DOXY-IVM groups showed a better result with a gradual decrease in microfilariae with no recurrence. These 2 groups were not only virtually deprived of infection but also sustained the sterility of infection through the course of study. These results revealed the advantages of using in B. malayi treatment in cats. Doxycycline showed to have both microfilaricidal and adulticidal effects on lymphatic filariae which maintained the long-term response to control of B. malayi infection in cats.

Human innate lymphoid cells (ILCs) in filarial infections.

Filarial infections are characteristically chronic and can cause debilitating diseases governed by parasite-induced innate and adaptive immune responses. Filarial parasites traverse or establish niches in the skin (migrating infective larvae), in nonmucosal tissues (adult parasite niche) and in the blood or skin (circulating microfilariae) where they intersect with the host immune response. While several studies have demonstrated that filarial parasites and their antigens can modulate myeloid cells (monocyte, macrophage and dendritic cell subsets), T- and B-lymphocytes and skin resident cell populations, the role of innate lymphoid cells during filarial infections has only recently emerged. Despite the identification and characterization of innate lymphoid cells (ILCs) in murine helminth infections, little is actually known about the role of human ILCs during parasitic infections. The focus of this review will be to highlight the composition of ILCs in the skin, lymphatics and blood; where the host-parasite interaction is well-defined and to examine the role of ILCs during filarial infections.

Impact of the Lymphatic Filariasis Control Program towards elimination of filariasis in Vanuatu, 1997\u20132006

BackgroundLymphatic filariasis (LF) occurs when filarial parasites are transmitted to humans through mosquitoes. The filarial worms affect the lymphatic system which leads to abnormal enlargement of body parts, chronic pain, disability, and social discrimination. In 1999, a commitment was made to eliminate LF from the Pacific Region by 2010. The Pacific Program to Eliminate LF began, with Vanuatu being one of the 16 endemic countries included in this program.MethodsIn 1997/1998 a LF prevalence baseline survey was conducted to determine the need for mass drug administration (MDA) in Vanuatu. In 1999, the Vanuatu Lymphatic Filariasis Control Program was established, and nationwide MDA was implemented from 2000 to 2004. LF prevalence was collected during the MDA through sentinel site and spot check surveys, and after 5 years of MDA. MDA implementation methods included health worker training, social mobilization, and culturally appropriate health promotion strategies.ResultsLF prevalence at baseline was 4.79%; after MDA this declined to 0.16% in 2005/2006. Average MDA coverage ranged from 75.5–81.5% across 5 years. All three evaluation units surveyed in 2005/2006 were below the 1% threshold required to stop MDA.ConclusionsThe LF Control Program between 1997 and 2006 was successful in reducing LF prevalence to <1%. High MDA coverage was a critical component of this success. This period of the Vanuatu LF Control Program played an important role in helping to eliminate LF in Vanuatu.

Molecular cloning and characterization of protein disulfide isomerase of Brugia malayi, a human lymphatic filarial parasite.

Lymphatic filariasis results in an altered lymphatic system and the abnormal enlargement of body parts, causing pain, serious disability and social stigma. Effective vaccines are still not available nowadays, drugs against the disease is required. Protein disulfide isomerase (PDI) is an essential catalyst of the endoplasmic reticulum which is involved in folding and chaperone activities in different biological systems. Here, we report the enzymatic characterization of a Brugia malayi Protein disulfide isomerase (BmPDI), which was expressed and purified from Escherichia coli BL21 (DE3). Western blotting analysis showed the recombinant BmPDI could be recognized by anti-BmPDI Rabbit serum. The rBmPDI exhibited an optimum activity at pH 8 and 40 °C. The enzyme was inhibited by aurin and PDI inhibitor. Recombinant BmPDI showed interaction with recombinant Brugia malayi calreticulin (rBmCRT). The three-dimensional model for BmPDI and BmCRT was generated by homology modelling. A total of 25 hydrogen bonds were found to be formed between two interfaces. There are 259 non-bonded contacts present in the BmPDI-BmCRT complex and 12 salt bridges were formed in the interaction.

Rapid integrated clinical survey to determine prevalence and co-distribution patterns of lymphatic filariasis and onchocerciasis in a Loa loa co-endemic area: The Angolan experience

The Republic of Angola is a priority country for onchocerciasis and lymphatic filariasis (LF) elimination, however, the co-distribution of the filarial parasite Loa loa (loiasis) is a significant impediment, due to the risk of severe adverse events (SAEs) associated with ivermectin used in mass drug administration (MDA) campaigns. Angola has a high risk loiasis zone identified in Bengo Province where alternative interventions may need to be implemented; however, the presence and geographical overlap of the three filarial infections/diseases are not well defined. Therefore, this study conducted a rapid integrated filarial mapping survey based on readily identifiable clinical conditions of each disease in this risk zone to help determine prevalence and co-distribution patterns in a timely manner with limited resources. In total, 2007 individuals from 29 communities in five provincial municipalities were surveyed. Community prevalence estimates were determined by the rapid assessment procedure for loiasis (RAPLOA) and rapid epidemiological mapping of onchocerciasis (REMO) together with two questions on LF clinical manifestations (presence of lymphoedema, hydrocoele). Overall low levels of endemicity, with different overlapping distributions were found. Loiasis was found in 18 communities with a prevalence of 2.0% (31/1571), which contrasted to previous results defining the area as a high risk zone. Onchocerciasis prevalence was 5.3% (49/922) in eight communities, and LF prevalence was 0.4% for lymphoedema (8/2007) and 2.6% for hydrocoeles (20/761 males) in seven and 12 communities respectively. The clinical mapping survey method helped to highlight that all three filarial infections are present in this zone of Bengo Province. However, the significant difference in loiasis prevalence found between the past and this current survey suggests that further studies including serological and parasitological confirmation are required. This will help determine levels of infection and risk, understand the associations between clinical, serological and parasitological prevalence patterns, and better determine the most appropriate treatment strategies to reach onchocerciasis and LF elimination targets in the loiasis co-endemic areas. Our results also suggest that the utility of the earlier RAPLOA derived maps, based on surveys undertaken over a decade ago, are likely to be invalid given the extent of population movement and environmental change, particularly deforestation, and that fine scale micro-mapping is required to more precisely delineate the interventions required defined by these complex co-endemicities.

Anti-Onchocerca and Anti-Caenorhabditis Activity of a Hydro-Alcoholic Extract from the Fruits of Acacia nilotica and Some Proanthocyanidin Derivatives.

Acacia nilotica fruits with high tannin content are used in the northern parts of Cameroon as anti-filarial remedies by traditional healers. In this study, the hydro-alcoholic fruit extract (crude extract (CE)) and, one of the main constituents in its most active fractions, (+)-catechin-3-O-gallate (CG), as well as four related proanthocyanidins, (−)-epicatechin-3-O-gallate (ECG), (+)-gallocatechin (GC), (−)-epigallocatechin (EGC) and (−)-epigallocatechin-3-O-gallate (EGCG), were assessed for their potential in vitro anthelmintic properties against the free-living model organism Caenorhabditis elegans and against the cattle filarial parasite Onchocerca ochengi. Worms were incubated in the presence of different concentrations of fruit extract, fractions and pure compounds. The effects on mortality were monitored after 48 h. The plant extract and all of the pure tested compounds were active against O. ochengi (LC50 ranging from 1.2 to 11.5 µg/mL on males) and C. elegans (LC50 ranging from 33.8 to 350 µg/mL on wild type). While high LC50 were required for the effects of the compounds on C. elegans, very low LC50 were required against O. ochengi. Importantly, tests for acute oral toxicity (lowest dose: 10 mg/kg) in Wistar rats demonstrated that crude extract and pure compounds were non-toxic and safe to use. Additionally, the results of cytotoxicity tests with the Caco-2 cell line (CC50 ranging from 47.1 to 93.2 µg/mL) confirmed the absence of significant toxicity of the crude extract and pure compounds. These results are in good accordance with the use of A. nilotica against nematode infections by traditional healers, herdsmen and pastoralists in Cameroon.

Chronic Litomosoides sigmodontis infection protects against IgE-mediated anaphylaxis in sensitized mice

Abstract While numerous animal studies have reported that helminths can prevent allergic sensitization, few have evaluated the effects of infection on pre-existing allergy. We hypothesized that chronic infection would protect against the clinical symptoms of IgE-mediated anaphylaxis in previously sensitized mice. Mice were sensitized by weekly intraperitoneal (IP) injection of either ovalbumin (OVA)/alum or PBS/alum for 3 weeks. At 5 weeks, mice were infected with Litomosoides sigmodontis (L.s.), a rodent filarial parasite, or mock infected. Mice were challenged IP with OVA during the twelfth week of infection. Following challenge, serum levels of murine mast cell protease 1 (mMCP-1) were significantly lower in sensitized mice that were L.s.-infected as compared to mock infected (212,923 vs. 447,062 pg/mL, p-value= 0.0459). Additionally, L.s.-infected mice that had been previously sensitized exhibited an average drop in core body temperature of 3.5 °C after 30 min, which was significantly less than the 5.0 °C drop observed in sensitized and mock-infected mice. Anaphylaxis was IgE and mast-cell dependent, as mice deficient in FcɛRI or mast cells mice released little to no mMCP-1 or exhibited minimal drops in core body temperature in response to challenge. Experiments to date suggest that changes in mast cell functionality were central to the protective effect. First, serum levels of OVA-IgE were not altered by L. sigmodontis infection. Second, purified mast cells from OVA-sensitized and infected mice released less histamine than OVA-sensitized and mock-infected mice in response to OVA. These results suggest that chronic helminth infection protects against anaphylaxis in part by decreasing allergen-driven mast cell degranulation.