Fecundity Of Female Worms Research Articles

Immunization with excretory-secretory molecules of intestinal nematodes induces antigen-specific protective memory Th2 cell responses.

Parasitic nematodes infect more than 1 billion people in the global south. The development of effective antihelminthic vaccines is a crucial tool for their future elimination. Protective immune responses to nematodes depend on Gata3+ Th2 cells, which can also be induced by nematode-released products. Whether these nematode products induce antigen-specific long-lived memory T cells and thereby confer protection against a challenge infection is not known yet. Hence, we set out to characterize the formation of memory Th2 cells induced by immunization with Heligmosomoides polygyrus excretory-secretory (HES) products, infection-induced versus immunization-induced recall responses to a challenge infection, and whether HES-induced memory T cells show protective properties following adoptive transfer. Our results show that 8 weeks postimmunization, HES induces long-lived functional memory Th2 cells at the site of immunization in the peritoneal cavity. Following a H. polygyrus challenge infection, HES-immunized mice display MHC-II-dependent antigen-specific Th2 cytokine responses in the gut-draining lymph nodes, comparable to those induced by a prior natural infection. Moreover, adoptive transfer of sorted memory CD4+ T cells from HES-immunized donors reduces female worm fecundity following a challenge H. polygyrus infection in recipient mice, highlighting a protective role for immunization-induced memory T cells.

Co-Administration of Adjuvanted Recombinant Ov-103 and Ov-RAL-2 Vaccines Confer Protection against Natural Challenge in A Bovine Onchocerca ochengi Infection Model of Human Onchocerciasis.

Onchocerciasis (river blindness), caused by the filarial nematode Onchocerca volvulus, is a neglected tropical disease mainly of sub-Saharan Africa. Worldwide, an estimated 20.9 million individuals live with infection and a further 205 million are at risk of disease. Current control methods rely on mass drug administration of ivermectin to kill microfilariae and inhibit female worm fecundity. The identification and development of efficacious vaccines as complementary preventive tools to support ongoing elimination efforts are therefore an important objective of onchocerciasis research. We evaluated the protective effects of co-administering leading O. volvulus-derived recombinant vaccine candidates (Ov-103 and Ov-RAL-2) with subsequent natural exposure to the closely related cattle parasite Onchocerca ochengi. Over a 24-month exposure period, vaccinated calves (n = 11) were shown to acquire infection and microfilaridermia at a significantly lower rate compared to unvaccinated control animals (n = 10). Furthermore, adult female worm burdens were negatively correlated with anti-Ov-103 and Ov-RAL-2 IgG1 and IgG2 responses. Peptide arrays identified several Ov-103 and Ov-RAL-2-specific epitopes homologous to those identified as human B-cell and helper T-cell epitope candidates and by naturally-infected human subjects in previous studies. Overall, this study demonstrates co-administration of Ov-103 and Ov-RAL-2 with Montanide™ ISA 206 VG is highly immunogenic in cattle, conferring partial protection against natural challenge with O. ochengi. The strong, antigen-specific IgG1 and IgG2 responses associated with vaccine-induced protection are highly suggestive of a mixed Th1/Th2 associated antibody responses. Collectively, this evidence suggests vaccine formulations for human onchocerciasis should aim to elicit similarly balanced Th1/Th2 immune responses.

Pyrvinium Pamoate and Structural Analogs Are Early Macrofilaricide Leads.

Onchocerciasis and lymphatic filariasis are neglected tropical diseases caused by infection with filarial worms. Annual or biannual mass drug administration with microfilaricidal drugs that kill the microfilarial stages of the parasites has helped reduce infection rates and thus prevent transmission of both infections. However, success depends on high population coverage that is maintained for the duration of the adult worm’s lifespan. Given that these filarial worms can live up to 14 years in their human hosts, a macrofilaricidal drug would vastly accelerate elimination efforts. Here, we have evaluated the repurposed drug pyrvinium pamoate as well as newly synthesized analogs of pyrvinium for their efficacy against filarial worms in vitro and in vivo. We found that pyrvinium pamoate, tetrahydropyrvinium and one of the analogs were highly potent in inhibiting worms in in vitro whole-worm screening assays, and that all three compounds reduced female worm fecundity and inhibited embryogenesis in the Brugia pahangi-gerbil in vivo model of infection.

Comparative Study of Laser- attenuated and Gamma attenuated irradiated larval Vaccines of Dictyocaulus filaria in goats.

A comparative trial between the effect of 3rd stage Dictyocaulus filaria larval vaccine attenuated by continuous emission of visible Helium – Neon Laser of ImW and wave length of ( 632.8 ) nm of (5) minutes exposure that of gamma attenuated larvae at 0.5 k. gray ( Co as radiation Source ) was designed. Each of the two attenuated larval vaccines was given to a group of ( 5 ) kids each . Double immunization doses at ( 35 ) days interval from each vaccine were given orally. The first dose contained ( 1000 ) larvae and the second one ( 2000 ) for each . A challenge dose of ( 100 ) non- irradiated larvae /kg body weight was given after ( 5 ) week to the vaccinated groups and a control non- vaccinated (5 kids ) 3rd group. All animals were slaughtered after 6 weeks post challenge. 
 Results showed that both vaccines revealed 80.7% and 78.1% protection respectively for worm burden as compared to control. Laser attenuated vaccine exhibited a statistically significant inhibition in the fecundity of female worms and larvae secretion in faeces in comparison with the gamma irradiated vaccine.

Emodepside targets SLO-1 channels of Onchocerca ochengi and induces broad anthelmintic effects in a bovine model of onchocerciasis.

Onchocerciasis (river blindness), caused by the filarial worm Onchocerca volvulus, is a neglected tropical disease mostly affecting sub-Saharan Africa and is responsible for >1.3 million years lived with disability. Current control relies almost entirely on ivermectin, which suppresses symptoms caused by the first-stage larvae (microfilariae) but does not kill the long-lived adults. Here, we evaluated emodepside, a semi-synthetic cyclooctadepsipeptide registered for deworming applications in companion animals, for activity against adult filariae (i.e., as a macrofilaricide). We demonstrate the equivalence of emodepside activity on SLO-1 potassium channels in Onchocerca volvulus and Onchocerca ochengi, its sister species from cattle. Evaluation of emodepside in cattle as single or 7-day treatments at two doses (0.15 and 0.75 mg/kg) revealed rapid activity against microfilariae, prolonged suppression of female worm fecundity, and macrofilaricidal effects by 18 months post treatment. The drug was well tolerated, causing only transiently increased blood glucose. Female adult worms were mostly paralyzed; however, some retained metabolic activity even in the multiple high-dose group. These data support ongoing clinical development of emodepside to treat river blindness.

Feeding of carob (Ceratonia siliqua) to sheep infected with gastrointestinal nematodes reduces faecal egg counts and worm fecundity

The present study explored the anthelmintic effects of condensed tannins (CT) in carob (Ceratonia siliqua) pods fed to sheep against gastrointestinal nematodes. Three independent in vivo trials tested whether i) carob pod (CaBP)-containing feed had an anthelmintic effect and if yes, which was the optimal concentration in the diet; ii) whether this effect could be attributed to tannins through the polyethylene glycol (PEG) test and iii) whether there were any synergistic effects when combined with another tannin-containing feed (e.g. sainfoin). In all trials 6-month old nematode-naive lambs, experimentally infected with both Haemonchus contortus and Trichostrongylus colubriformis, were used. Faecal egg counts (FEC) were performed regularly and at the end of each trial adult worm counts (AWC) and female worm fecundity were recorded. In trial 1, 35 lambs (five groups of seven lambs) were fed different CaBP concentrations ranging from 0% to 12 % w/w. FEC declined up to 39.2 % only in the group fed with 12 %CaBP, while a declining trend (P < 0.06) was demonstrated for the AWC of T. colubriformis, which was associated with the increasing concentration of CaBP in feed. Female worm fecundity was reduced in groups fed CaBP for both parasites, however this was only significant for H. contortus (P < 0.001), in a dose dependent manner. In trial 2, four groups of six infected lambs each were used, which received the carob diets CaBP or CaBP + PEG, and the tannin-free diets with or without PEG (C or C + PEG). Results showed that FEC of Groups C, C + PEG, and CaBP + PEG were comparable throughout the trial, while the group receiving only CaBP showed lower FEC from DAY 25 onwards. AWC showed a reduction (67.7 %) only for H. contortus (P < 0.03). Reversal of the anthelmintic effect of CaBP after PEG administration suggested that CT contributed to the anthelmintic action. However, no effect of CaBP was observed on T. colubriformis AWC and on female worm fecundity for both species. Finally, for trial 3 four groups of six lambs each received a diet based on CaBP, sainfoin (S) or a combination (CaBP + S) and were compared to a control (C) diet of lucerne. On DAY 37 FEC values in groups CaBP + S and S tended to be lower compared to the two other groups (C, CaBP), while for AWCs no significant differences were observed for both parasites. The fecundity of H. contortus and T. colubriformis demonstrated significant differences between the treated and control groups, with lower values in the animals receiving CaBP + S. Overall, the results supported the hypothesis that carob had an anthelmintic effect due to its CT, but there was no clear indication of a synergistic effect with sainfoin.

The endosymbiont Wolbachiarebounds following antibiotic treatment.

Antibiotic treatment has emerged as a promising strategy to sterilize and kill filarial nematodes due to their dependence on their endosymbiotic bacteria, Wolbachia. Several studies have shown that novel and FDA-approved antibiotics are efficacious at depleting the filarial nematodes of their endosymbiont, thus reducing female fecundity. However, it remains unclear if antibiotics can permanently deplete Wolbachia and cause sterility for the lifespan of the adult worms. Concerns about resistance arising from mass drug administration necessitate a careful exploration of potential Wolbachia recrudescence. In the present study, we investigated the long-term effects of the FDA-approved antibiotic, rifampicin, in the Brugia pahangi jird model of infection. Initially, rifampicin treatment depleted Wolbachia in adult worms and simultaneously impaired female worm fecundity. However, during an 8-month washout period, Wolbachia titers rebounded and embryogenesis returned to normal. Genome sequence analyses of Wolbachia revealed that despite the population bottleneck and recovery, no genetic changes occurred that could account for the rebound. Clusters of densely packed Wolbachia within the worm’s ovarian tissues were observed by confocal microscopy and remained in worms treated with rifampicin, suggesting that they may serve as privileged sites that allow Wolbachia to persist in worms while treated with antibiotic. To our knowledge, these clusters have not been previously described and may be the source of the Wolbachia rebound.

Fecundity of adult female worms were affected when Brugia malayi infected Mongolian gerbils were immunized with a multivalent vaccine (rBmHAXT) against human lymphatic filarial parasite

A multivalent recombinant fusion protein prophylactic vaccine, rBmHAXT developed against lymphatic filariasis (LF) demonstrated over 57% protection against challenge infection in rhesus macaque model. Currently, we do not know if the rBmHAXT vaccination has any effect on adult worms and/or on the fecundity of adult female worms. Thus, the major focus of this study was to determine the effect of rBmHAXT vaccination on Brugia malayi infected mongolian gerbils. We performed two sets of experiments. In the first set of experiment, gerbils were infected with 100 B. malayi L3. After confirming the establishment of infection, four rounds of DEC treatment and rBmHAXT vaccination was given. Results showed that following vaccination with rBmHAXT, the microfilaria (Mf) count was significantly decreased in all vaccinated animals compared to controls. At the end of these experiments, we collected and counted the established adult worms. There was a 36% reduction in the recovery of adult female worms, which might account for the low Mf load in vaccinated animals. In the second set of experiments, animals were vaccinated first with rBmHAXT followed by surgically implanting adult male or female B. malayi parasites into the peritoneal cavity to determine the effect of vaccination on each sex of the parasite. Our results show that the rBmHAXT vaccination has no effect on male adult worms compared to controls. However, there was 40% reduction in the Mf load in vaccinated animals that were transplanted with adult female worms. These findings suggested that the rBmHAXT vaccination has potential damaging effect on the fecundity of adult female worms. Scanning electron microscopy studies showed cuticular damage on the surface of adult female worms. These studies thus show that the rBmHAXT vaccination in infected gerbils has partial microfilaricidal effect and potentially affect the fecundity of adult female worms.

Evaluation of the in vivo anthelmintic properties of Mitragyna inermis (Willd.) as a livestock dewormer against parasitic hematophagous worm Haemonchus contortus infections in different breeds of lambs.

Gastrointestinal (GI) nematodes remain a major constraint on livestock production throughout the world. This study assessed the in vivo efficacy of the powder of Mitragyna inermis leaves in three breeds of lambs, namely, West African dwarf lambs (WAD), West African long-legged lambs (WALL), and F1 lambs (cross of a WALL ram with a WAD ewe), artificially infected with 3000 L3s of Haemonchus contortus in a controlled experiment. Fecal sample examination, serological analysis, and necropsy were carried out to determine the egg count, worm burden, and worm fecundity reduction. A dose of 3.2g/kg body weight (BW) M. inermis was administered per the oral route for three consecutive days and repeated 2weeks later. Compared with the control, the powder of M. inermis leaves (> 60%) and albendazole (100%) significantly reduced (p < 0.01) fecal egg counts (FECs) in the three breeds of lambs. The posttreatment reduction in FECs fluctuated from 56.99 to 78.75% for WAD lambs, 38.39 to 66.39% for WALL lambs, and 35.55 to 63.11% for F1 lambs (WALL × WAD). Significant differences (p < 0.05) were observed in packed cell volume values before and after infection. M. inermis reduced the egg-laying capacity of female adult worms by up to 60% and eliminated more than 80% of the adult worms of H. contortus in lambs. Furthermore, albendazole reduced the worm count and fecundity of female worms by greater amounts than M. inermis (100%). The findings of this study showed that M. inermis is a good source of bioactive compounds for drug development. According to this result, a 3.2g/kg BW dose of the plant could be applied for the control of GI nematodes in small ruminants.

Variation in faecal worm egg counts of experimentally infected goats and mice with time of day and its implications in diagnosis of helminthosis.

Faecal worm egg counts (FEC) are very important indicators in assessing the intensity of helminth infections in animal hosts and inform decisions taken in parasite control programmes. FEC are however affected by many factors which include the dose level of larval challenge, species composition of the worms, worm burden, female worm fecundity and concurrent infections to name but these few. The relevance of time of collection of faecal sample for FEC is not fully known and in most cases faeces for analysis is collected whenever feasible for the researcher on sample collection days. The significance of the time of collection of faeces on FEC was therefore investigated at two different periods of the day, morning and afternoon. Faecal samples were collected at 7-8a.m. (morning) and at 2-3p.m. (afternoon) on same sampling days from 6 mice and 10 West African Dwarf goat kids experimentally infected with Heligmosomoides bakeri and Haemonchus contortus respectively. FEC were conducted using the flotation and modified McMaster techniques. Overall, the 2-3p.m. FEC tended to be higher than the 7-8a.m. FEC in both animal species but the difference was not significant (P>0.05). The time of collection of faecal samples for faecal worm egg counts is therefore not a crucial factor that may significantly affect FEC in H. bakeri and H. contortus infected mice and goats respectively although it would be advisable to maintain particular periods for collection of faeces in particular experiments for best results of FEC to be obtained.

Meta-analysis of the parasitic phase traits of Haemonchus contortus infection in sheep

BackgroundThe parasitic nematode Haemonchus contortus shows highly variable life history traits. This highlights the need to have an average estimate and a quantification of the variation around it to calibrate epidemiological models.MethodsThis paper aimed to quantify the main life history traits of H. contortus and to identify explanatory factors affecting these traits using a powerful method based on a systematic review and meta-analysis of current literature. The life history traits considered are: (i) the establishment rate of ingested larvae; (ii) the adult mortality rate; (iii) the fertility (i.e. the number of eggs laid/female/day); and (iv) fecundity of female worms (i.e. the number of eggs per gram of faeces).ResultsA total of 37 papers that report single experimental infection with H. contortus in sheep and published from 1960 to 2015, were reviewed and collated in this meta-analysis. This encompassed 115 experiments on 982 animals. Each trait was analysed using a linear model weighted by its inverse variance. The average (± SE) larval establishment rate was 0.24 ± 0.02, which decreased as a function of the infection dose and host age. An average adult mortality rate of 0.021 ± 0.002) was estimated from the literature. This trait varied as a function of animal age, breed and protective response due to prior exposure to the parasite. Average female fertility was 1295.9 ± 280.4 eggs/female/day and decreased in resistant breeds and previously infected hosts. Average faecal egg count at necropsy was 908.5 ± 487.1 eggs per gram of faeces and varied as a function of infection duration and host resistance. The average sex ratio of H. contortus was 0.51 ± 0.006.ConclusionThis work is the first systematic review to summarise the available information on the parasitic phase of H. contortus in sheep. The results of the meta-analysis provide robust estimates of life history traits for parametrization of epidemiological models, their expected variation according to experimental factors, and provides correlations between these.

Modulation of Haemonchus contortus infection by depletion of γδ+ T cells in parasite resistant Canaria Hair Breed sheep

Canaria Hair Breed (CHB) sheep display resistance against the adult stage of the nematode, Haemonchus contortus. Previous studies have suggested significant correlations between γδ+ T lymphocytes and fecundity of female adult worms, suggesting a novel role in immune modulation by these cells. The largest proportion of γδ+ T lymphocytes in sheep are the subpopulation of γδ+/WC1+ T cells. The aim of this study was to evaluate the effect of γδ+⁄WC1+ T cell depletion via infusion of anti-γδ/WC1 monoclonal antibody (mAb) on the subsequent immune response of CHB sheep infected with H. contortus. Significantly lower γδ+ T cell levels in both peripheral blood and in the basal layers of the abomasal tissue resulted following anti-γδ/WC1 mAb infusion of CHB sheep compared to control animals. Worms recovered from the anti-γδ/WC1 mAb treated CHB sheep had significantly longer female worms with correspondingly more eggs in utero than the saline control group. Significant correlations between eosinophils and worm length and fecundity were no longer apparent in the anti-γδ/WC1 mAb treated CHB sheep. These results support the notion that γδ+ T cells in CHB sheep play a critical role in fecundity regulation (length and eggs in utero) of H. contortus adult female worms, and highlights a new mechanism of modulation by this lymphocyte population, possibly involving eosinophil activation.

Intestinal helminth infection induces highly functional resident memory CD4+ T cells in mice.

Immunity to intestinal nematodes requires CD4⁺ Th2-cell responses, including IL-4 and IL-13 production. Chronic infection with intestinal nematodes leads to downregulation of these responses, and few functional T helper (Th) 2 cells are detected in secondary lymphoid organs in the chronic phase or after abrogation of infection. Here, we show with a natural murine infection with Heligmosomoides polygyrus that highly functional memory Th2 cells persist in the lamina propria and in addition in the peritoneal cavity (PC) after abrogation of infection. While both tissue-resident memory (TRM ) populations proliferate in situ and express IL-4, IL-5, and IL-13 upon TCR-dependent stimulation, only peritoneal memory cells express high levels of the IL-33 receptor and produce IL-5 and IL-13 upon TCR-independent stimulation with IL-33 and IL-7. Most importantly, PC-derived TRM cells are able to mediate anti-helminthic effects by decreasing the fecundity of female worms upon transfer into recipient mice. These results show that nonlymphoid compartments can serve as reservoirs for Th2 memory cells, and furthermore that innate effector function of Th2 memory cells is restricted to CD4⁺ memory T cells residing in the PC.

Vaccination of Gerbils with Bm-103 and Bm-RAL-2 Concurrently or as a Fusion Protein Confers Consistent and Improved Protection against Brugia malayi Infection.

BackgroundThe Brugia malayi Bm-103 and Bm-RAL-2 proteins are orthologous to Onchocerca volvulus Ov-103 and Ov-RAL-2, and which were selected as the best candidates for the development of an O. volvulus vaccine. The B. malayi gerbil model was used to confirm the efficacy of these Ov vaccine candidates on adult worms and to determine whether their combination is more efficacious.Methodology and Principle FindingsVaccine efficacy of recombinant Bm-103 and Bm-RAL-2 administered individually, concurrently or as a fusion protein were tested in gerbils using alum as adjuvant. Vaccination with Bm-103 resulted in worm reductions of 39%, 34% and 22% on 42, 120 and 150 days post infection (dpi), respectively, and vaccination with Bm-RAL-2 resulted in worm reductions of 42%, 22% and 46% on 42, 120 and 150 dpi, respectively. Vaccination with a fusion protein comprised of Bm-103 and Bm-RAL-2 resulted in improved efficacy with significant reduction of worm burden of 51% and 49% at 90 dpi, as did the concurrent vaccination with Bm-103 and Bm-RAL-2, with worm reduction of 61% and 56% at 90 dpi. Vaccination with Bm-103 and Bm-RAL-2 as a fusion protein or concurrently not only induced a significant worm reduction of 61% and 42%, respectively, at 150 dpi, but also significantly reduced the fecundity of female worms as determined by embryograms. Elevated levels of antigen-specific IgG were observed in all vaccinated gerbils. Serum from gerbils vaccinated with Bm-103 and Bm-RAL-2 individually, concurrently or as a fusion protein killed third stage larvae in vitro when combined with peritoneal exudate cells.ConclusionAlthough vaccination with Bm-103 and Bm-RAL-2 individually conferred protection against B. malayi infection in gerbils, a more consistent and enhanced protection was induced by vaccination with Bm-103 and Bm-RAL-2 fusion protein and when they were used concurrently. Further characterization and optimization of these filarial vaccines are warranted.

Faecal egg counts and expulsion dynamics of the whipworm, Trichuris trichiura following self-infection.

More than 400 million humans are estimated to be infected with the intestinal helminth parasite, Trichuris trichiura. The infection is chronic in nature and high-intensity infection can lead to colitis, anaemia, Trichuris Dysentery Syndrome and reduced cognitive performance. Single doses of 400mg albendazole or 500mg mebendazole (MBZ) are used in mass drug administration programmes, but this has been shown to be insufficient. In this study, worm expulsion dynamics are described after MBZ treatment, given as a multi-dose and single-dose treatment in two separate T. trichiura self-infection studies. Worm expulsion dynamics post-treatment showed a similar pattern regardless of the dose regime, with the first worms observed on day 2 and the last worms expelled on days 9 and 13 post-treatment. Establishment of a chronic infection was observed following the inefficient single-dose treatment. The prepatent period was 13-16 weeks in both studies and worms were found to have a lifespan of at least 1 year and 10 months. These self-infection studies provide key information on the chronicity of T. trichiura infections, expulsion dynamics after anthelmintic treatment and the prepatent period, as well as the fecundity of female worms, which was around 18,000 eggs/female per day.

Tolerance and resistance to a nematode challenge are not always mutually exclusive

The relationship between the manifestations of tolerance (a host’s ability to reduce the impact of a given level of pathogens) and resistance (a host’s ability to clear pathogens) has been assumed to be an antagonistic one. Here we tested the hypothesis that mice from strains more resistant to intestinal nematodes will experience reduced tolerance compared with less resistant mice. Three inbred strains of mice were used: C57BL/6 mice have been characterised as susceptible, whereas BALB/c and NIH mice have been characterised as resistant to Heligmosomoides bakeri infection. Mice of each strain were either parasitised with a single dose of 250 L3H. bakeri (n=10) in water or were sham-infected with water (n=10). Body weight, food intake and worm egg output were recorded regularly throughout the experiment. Forty-two days p.i. mice were euthanised and organ weights, eggs in colon and worm counts were determined. C57BL/6 mice showed significantly greater worm egg output (P<0.001), eggs in colon (P<0.05) and female worm fecundity (P<0.05) compared with NIH and BALB/c mice. Parasitised BALB/c mice grew more whilst parasitised C57BL/6 mice grew less than their sham-infected counterparts during the first 2weeks post-challenge (P=0.05). Parasitism significantly increased liver, spleen, small intestine and caecum weights (P<0.001) but reduced carcass weight (P<0.01). Average daily weight gain and worm numbers were positively correlated in NIH mice (P=0.05); however, the relationship was reversed when carcass weight was used as a measure for tolerance. BALB/c mice did not appear to suffer from the consequences of parasitism, with carcass weight similar in all animals. Our hypothesis that strains more resistant to the H. bakeri infection are less tolerant compared with less resistant strains is rejected, as the two resistant strains showed variable tolerance. Thus, tolerance and resistance to an intestinal nematode infection are not always mutually exclusive.

Immunogenicity and anti-fecundity effect of nanoparticle coated glutathione S-transferase (SjGST) DNA vaccine against murine Schistosoma japonicum infection

There is still urgent need for a vaccine against schistosomiasis, especially in Schistosoma japonicum endemic areas where even a vaccine that will interrupt zoonotic transmission will be potentially effective as an intervention tool. We had developed a novel nanoparticle gene delivery system, which has proven efficacious in gene transfection to target immune cells with complementary adjuvant effect and high protective efficacy in several diseases. Here, we applied this nanoparticle system in combination with S. japonicum glutathione S-transferase (SjGST) DNA vaccine to show the immunogenicity and anti-fecundity effect of the nanoparticle coated vaccine formulation against murine schistosomiasis. The nanoparticle-coated DNA vaccine formulation induced desired immune responses. In comparison with the nanoparticle coated empty vector, it produced significantly increased antigen-specific humoral response, T-helper 1 polarized cytokine environment, higher proportion of IFN-γ producing CD4+ T-cells and the concomitant decrease in IL-4 producing CD4+ T-cells. Although there was no effect on worm burden, we recorded a marked reduction in tissue egg burden. There was up to 71.3% decrease in tissue egg burden and 55% reduction in the fecundity of female adult worms. Our data showed that SjGST DNA vaccine, delivered using the nanoparticle gene delivery system, produced anti-fecundity effect on female adult schistosomes as previously described by using conventional subunit vaccine with adjuvant, proving this DNA vaccine formulation as a promising candidate for anti-pathology and transmission blocking application.

Anthelmintic effect of heather in goats experimentally infected with Trichostrongylus colubriformis

The effects of heather (composed primarily of Calluna vulgaris with a smaller content of Erica umbellata and Erica cinerea) consumption on the establishment of incoming infective larvae (experiment 1, preventive treatment) and an adult worm population (experiment 2, curative treatment) were investigated in Cashmere goats experimentally infected with Trichostrongylus colubriformis. In experiment 1, 12 castrated male goats were divided into two groups: heather-supplemented vs. non-supplemented animals. After 2 weeks of adaptation to the diet, all goats were experimentally infected per os with 6,000 T. colubriformis third-stage larvae. Three weeks post-infection, the goats were slaughtered, and worm counts as well as female worm fecundity and development were determined. Heather consumption was associated with a close to significant (P=0.092) reduction (mean 14 %) in larvae establishment. No effect on fecundity was observed, but the length of female worms in supplemented goats was greater (P<0.001). In experiment 2, 15 non-lactating does were experimentally infected with 6,000 T. colubriformis third-stage larvae. At 6 weeks post-infection, three groups were established: control, heather-supplemented and heather-supplemented with polyethylene glycol. Individual faecal nematode egg output was measured twice weekly to assess gastrointestinal nematode egg excretion. The goats were slaughtered 5 weeks after heather administration (11 weeks post-infection), and worm counts as well as female worm fecundity and development were subsequently determined. Heather administration was associated with a significant (P<0.001) decrease (between 47 and 66 % compared with control group) in egg excretion from 45 to 76 days post-infection. Although worm counts and female fecundity were lower in supplemented goats, no significant differences were observed. Overall, the results showed a reduction in T. colubriformis larvae establishment and a decrease in nematode egg excretion when heather was administered in experimentally infected goats. The heather plus polyethylene glycol treatment reduced nematode egg excretion levels at the same proportion as heather, thereby suggesting that the threshold of tannins required for an anthelmintic effect is most likely quite low.

Dynamics of Onchocerca volvulus Microfilarial Densities after Ivermectin Treatment in an Ivermectin-naïve and a Multiply Treated Population from Cameroon

Background/ObjectiveIvermectin has been the keystone of onchocerciasis control for the last 25 years. Sub-optimal responses to the drug have been reported in Ghanaian communities under long-term treatment. We assessed, in two Cameroonian foci, whether the microfilaricidal and/or embryostatic effects of ivermectin on Onchocerca volvulus have been altered after several years of drug pressure.MethodsWe compared the dynamics of O. volvulus skin microfilarial densities after ivermectin treatment in two cohorts with contrasting exposure to this drug: one received repeated treatment for 13 years whereas the other had no history of large-scale treatments (referred to as controls). Microfilarial densities were assessed 15, 80 and 180 days after ivermectin in 122 multiply treated and 127 ivermectin-naïve individuals. Comparisons were adjusted for individual factors related to microfilarial density: age and number of nodules.FindingsTwo weeks post ivermectin, microfilarial density dropped equally (98% reduction) in the ivermectin-naïve and multiply treated groups. Between 15 and 180 days post ivermectin, the proportion of individuals with skin microfilariae doubled (from 30.8% to 67.8%) in controls and quadrupled (from 19.8% to 76.9%) in multiply treated individuals but the mean densities remained low in both sites. In fact, between 15 and 80 days, the repopulation rate was significantly higher in the multiply treated individuals than in the controls but no such difference was demonstrated when extending the follow-up to 180 days. The repopulation rate by microfilariae was associated with host factors: negatively with age and positively with the number of nodules.ConclusionThese observations may indicate that the worms from the multi-treated area recover mf productivity earlier but would be less productive than the worms from the ivermectin-naïve area between 80 and 180 days after ivermectin. Moreover, they do not support the operation of a strong cumulative effect of repeated treatments on the fecundity of female worms as previously described.

Effect of the consumption of heather on incoming larvae and established population of Teladorsagia circumcincta in experimentally infected Cashmere goats

This study was performed in Cashmere goats that were experimentally infected with Teladorsagia circumcincta to investigate the effects of heather consumption on the establishment of incoming infective larvae (experiment 1) and on an adult nematode population (experiment 2). In experiment 1, 24 non-lactating goats were divided into 2 groups: heather-supplemented vs. non-supplemented. After 2 weeks of adaptation to the diet, all of the goats were experimentally infected with 6000 infective larvae of T. circumcincta. Twelve animals (6 controls and 6 supplemented with heather) were slaughtered at 6 days and at 3 weeks post-infection (pi). After slaughter, the worms were counted and the female worm fecundity and development were determined. Heather consumption was associated with a significant reduction in larval establishment at 6 days (P=0.033) and at 3 weeks (P=0.041) pi. No differences in worm counts between the slaughter times were found. In the goats slaughtered at 3 weeks pi, the number of eggs in utero and length of the female worms were significantly (P<0.001) lower than those of control group. In experiment 2, 24 non-lactating goats were experimentally infected with 10,000 T. circumcincta infective larvae daily for 5 consecutive days (total infection of 50,000 larvae). After 3 weeks, 2 groups were established: control and heather-supplemented. The faecal egg output of each animal was measured at 2-days interval during the experimental period. The goats were slaughtered after 3 weeks of heather administration for parasitological studies. Heather administration was associated with a significant decrease in egg excretion between 25 and 29 days pi. The worm counts were similar in both groups, but the female length and fecundity were significantly (P<0.001) lower in supplemented goats. These results show that heather consumption reduces the establishment of T. circumcincta larvae in goats and the development and fecundity of female adult parasites.