Haematophagous Nematode Research Articles

Host-Parasite Interaction Between the Nematoda Procamallanus (Spirocamallanus) paraensis and Fish Hoplias malabaricus From the Natural Environment in the Brazilian Amazon.

Histopathological studies of parasitic infections in fish from the natural environment of Brazilian Amazon, are quite scarce. In this study, we investigated the histopathological changes of the proximal intestine of specimens of the Amazonian fish Hoplias malabaricus infected by the hematophagous nematode Procamallanus (Spirocamallanus) paraensis. The prevalence of the infection was 60%, with an average abundance of 1.46 and an average intensity of 2.43 parasites/fish. No significant difference was observed in the prevalence of infection and fish size or sex, but larger fish showed greater infection intensity, which was also significantly higher in male hosts. Histological sections of the proximal intestine showed reduction and loss of the epithelial lining, exposure of the lamina propria where the nematode interacts with the intestine wall, through the insertion of the buccal capsule and fish cellular debris in the intestinal lumen. In addition, areas with bleeding and inflammatory infiltrate were observed, but no changes or presence of parasite structures were observed in the other tunics of the intestinal wall.

In vivo anthelmintic activity of Combretum mucronatum schumach & Thonn leaf extract against Haemonchus contortus in goats

Parasitic infections with gastrointestinal nematodes are a serious problem for the health and welfare of domestic animals and negatively affect the economics of animal production. Haemonchus contortus is a haematophagous nematode of small ruminants responsible for significant mortality and morbidity. In addition, the widespread resistance to synthetic anthelmintic drugs emphasizes the urgent need of alternative treatment options against haemonchosis. This work aims to investigate the anthelmintic activity of an hydroethanolic Combretum mucronatum leaf extract (CMLE) against Haemonchus contortus in goats. Goats were artificially infected with 3500 third-stage larvae of H. contortus, and 21 days later, treated with CMLE (1000, 500, 250 mg/kg) for 4 consecutive days. Different parameters such as faecal egg count reduction, weight and haematocrit were monitored during the experimental period. The number of eggs per gram of faeces (EPG) was concentration-depended lower and significantly reduced compared to the untreated control (p < 0.0001). The effect of the highest CMLE dose (4 ×1000 mg/kg body weight) was similar to the effect of albendazole (1 ×5 mg/kg of body weight). The ED50 and ED90 values calculated were 189.17 and 392.33 mg/kg body weight respectively. ED50 and ED90 values were time-dependent. Moreover, CMLE improved haematocrit and weight of goats in dose-dependent and time-dependent manner. These results showed that CMLE could be used for haemonchosis treatment in small ruminants.

Real-time single-base specific detection of the Haemonchus contortus S168T variant associated with levamisole resistance using loop-primer endonuclease cleavage loop-mediated isothermal amplification

Haemonchus contortus is a parasitic haematophagous nematode that primarily affects small ruminants and causes significant economic loss to the global livestock industry. Treatment of haemonchosis typically relies on broad-spectrum anthelmintics, resistance to which is an important cause of treatment failure. Resistance to levamisole remains less widespread than to other major anthelmintic classes, prompting the need for more effective and accurate surveillance to maintain its efficacy. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) is a recently developed diagnostic method that facilitates multiplex target detection with single nucleotide polymorphism (SNP) specificity and portable onsite testing. In this study, we designed a new LEC-LAMP assay and applied it to detect the levamisole resistance marker S168T in H. contortus. We explored multiplexing probes for both the resistant S168T and the susceptible S168 alleles in a single-tube assay. We then included a generic probe to detect the acr-8 gene in the multiplex assay, which could facilitate the quantification of both resistance markers and overall genetic material from H. contortus in a single step. Our results showed promising application of these technologies, demonstrating a proof-of-concept assay which is amenable to detection of resistance alleles within the parasite population, with the potential for multiplex detection, and point-of-care application enabled by lateral flow end-point detection. However, further optimisation and validation is necessary.

Haemonchosis in Sheep and Goats, Control Strategies and Development of Vaccines against Haemonchus contortus.

Simple SummaryHaemonchus contortus is the most pathogenic blood-feeding parasitic nematode in sheep and goats, threatening animal welfare and causing tremendous economic losses to the small ruminant industry. This comprehensive review article sums up current control strategies, worm-derived antigens and recent advances in anti-Haemonchus vaccine development. New insights into antigen engineering and general considerations for clinical trials are discussed here.The evolutionary success of parasitic worms causes significant economic losses and animal health problems, including in the small ruminant industry. The hematophagous nematode Haemonchus contortus is a common endoparasite that infects wild and domestic ruminants worldwide, especially in tropical and subtropical regions. To date, the most commonly applied control strategy is the administration of anthelminthic drugs. The main disadvantages of these chemicals are their ecotoxic effects, the necessary withdrawal period (especially important in dairy animals) and the increasing development of resistance. Vaccines offer an attractive alternative control strategy against Haemonchus infections. In previous years, several potential vaccine antigens prepared from H. contortus using the latest technologies have been assessed in clinical trials using different methods and strategies. This review highlights the current state of knowledge on anti-H. contortus vaccines (covering native, recombinant and DNA-based vaccines), including an evaluation, as well a discussion of the challenges and achievements in developing protective, efficient, and long-lasting vaccines to control H. contortus infection and haemonchosis in small ruminants. This paper also addresses novel developments tackling the challenge of glycosylation of putative candidates in recombinant form.

In vitro effect of three tropical plants on adult Haemonchus placei, an haematophagous nematode from cattle

BackgroundVernonia amygdalina (leaf), Garcinia kola (seed), and Leucaena leucocephala (seed) are three well-known tropical plants used in African ethnomedicine to reduce parasitic worm burdens and are potential sources of alternative solution for controlling parasitic helminths infection in grazing livestock. This study investigated extracts from these plants for anthelmintic activity against adult Haemonchus placei, an haematophagous nematode from cattle abomasa. Powdered plant materials were macerated in acetone and the crude acetone extracts evaluated for anthelmintic activity using H. placei adult worm motility assay. Afterwards, fresh sample of V. amygdalina was macerated successively in chloroform and acetone and the extracts evaluated for anthelmintic activity. The chloroform extract was subjected to phytochemical and FT-IR analyses and fractionated by vacuum liquid chromatography. Anthelmintic data were fitted to a nonlinear regression equation (Log [extract or fraction] vs. lethality; variable slope) to produce best-fit sigmoidal curves and LC50 values computed with associated uncertainty.ResultsOf the three tropical plants, only V. amygdalina was active against adult H. placei with best-fit LC50 of 6.51 mg/mL (95% CI: 5.32–7.75). Evaluation of the two extracts obtained by successive maceration showed that chloroform extract (LC50, 2.46 mg/mL, 95% CI: 1.87–3.28) was 11 times as potent as acetone extract (LC50, 27.01 mg/mL, 95% CI: 21.32–48.57) (α < 0.0001). Chromatographic fractionation of the chloroform extract yielded four fractions (FA-FD) with FB (LC50, 2.38 mg/mL, 95% CI: 1.76–3.28) 2.19 times as potent as FC (LC50, 5.21 mg/mL, 95% CI: 4.40–5.79) against H. placei, while FA and FD were inactive. Phytochemical evaluation of the chloroform extract revealed the presence of saponins, steroids, terpenoids, cardiac glycosides, and the absence of tannins, flavonoids, alkaloids, and anthraquinones. FT-IR structural analysis of chloroform extract indicated the presence of key functional groups which are chemical fragments/ structural motifs known to be present in the two major classes of bioactive compounds (sesquiterpene lactones and steroid glucosides) reportedly to be found in V. amygdalina.ConclusionsThe findings showed that chloroform extract of V. amygdalina leaf possessed relatively good anthelmintic activity against adult H. placei. This could be indicative of its potential usefulness as an anthelmintic phytomedicine to control gastrointestinal nematodes infection in cattle.Key highlightsExtracts of three different plant materials (one leaf, two seeds) were tested against adult Haemonchus placei in vitro;Chloroform extract of Vernonia amygdalina was 11 times as potent as acetone extract;Fractionation of the chloroform extract yielded a bioactive fraction responsible for about 90% of the total lethal effect of the chloroform extract.Bioprocessing of V. amygdalina leaf could produce phytomedicines for organic livestock farming.Graphical abstract

Long-term ecological analysis of Anguillicola crassus occurrence and impact on the European eel population in a Mediterranean lagoon (North Italy)

Anguillicola crassus, a haematophagous nematode infecting the eel swim bladder, was introduced into Europe from Asia in the early 1980s with imported Anguilla japonica. It spread rapidly after establishment in the native European eel Anguilla anguilla. A long-term dataset shows the dynamics of A. crassus in eels of the Comacchio Lagoon from 1997 to 2019, and reveals the factors that influence its prevalence. A sample of 339 eels showed 19 parasitized by adult A. crassus, with one to five parasites per infected host and a mean abundance of 0.09 ± 0.45. Prevalence ranged from 2.14% (minimum value reached in 2005/06) to 11.9% (maximum reached in 1997). No significant temporal trend in the occurrence of A. crassus was detected over the period of the investigation, which was characterized by low levels of infection compared to that in eel populations from other coastal lagoons or estuaries. No influence of A. crassus on condition factor, hepatosomatic and gonadosomatic index and swim bladder integrity (evaluated through the Swim bladder Degenerative Index) was found, suggesting a minimal impact of the parasite on the eel population health. Anguillicola crassus, although established in Comacchio Lagoon, has not become invasive. The possible reasons for this, among which high salinity, will be discussed. Management of the lagoon for the past 30 years has also contributed to parasite containment through avoidance of restocking the eel population from external sources.

The contrasting hidden consequences of parasitism: Effects of a hematophagous nematode (Uncinaria sp.) in the development of a marine mammal swimming behavior.

Parasites are an important part of ecosystems, playing a critical role in their equilibrium. However, the consequences of parasitism beyond the direct effects associated with disease and mortality are not completely understood. This gap in knowledge is in part due to the difficulties to isolate the effect of single parasite species on physiological and behavioral traits in natural systems.The South American fur seal (Arctocephalus australis)–hookworm (Uncinaria sp.) interaction offers an ideal system to overcome these difficulties and study the behavioral and physiological effects of parasites in their hosts.Hookworms cause stunted growth and anemia in pinniped pups, which could affect early life active behaviors such as swimming. The aim of this study was to determine the effects of hookworms (Uncinaria sp.) on the development of swimming capabilities in A. australis through physiological and ethological analyses.Higher parasite burden was associated with reduced growth rates and lower blood hemoglobin concentrations, whereas scaled body mass and blood hemoglobin levels had an important positive effect on the water activity of the pups. However, antihookworm treatment did not affect the level of water activity of the pups, and pups with high hookworm burden increased their time budget in water. This was probably related to lower maternal attendance in heavily parasitized pups, leaving these pups more time to perform water activities. Therefore, pups with heavy hookworm burden, despite having decreased growth rates and blood hemoglobin concentrations, compensated for their handicap in physiological traits related to swimming by spending more time in the water.This work offers new insights to understand the contrasting effects of parasites on aquatic organisms, and the compensatory mechanisms employed by infected animals to avoid the worst consequences of parasitism.

Haematophagic Caenorhabditis elegans.

Caenorhabditis elegans is a free-living nematode that resides in soil and typically feeds on bacteria. We postulate that haematophagic C. elegans could provide a model to evaluate vaccine responses to intestinal proteins from hematophagous nematode parasites, such as Necator americanus. Human erythrocytes, fluorescently labelled with tetramethylrhodamine succinimidyl ester, demonstrated a stable bright emission and facilitated visualization of feeding events with fluorescent microscopy. C. elegans were observed feeding on erythrocytes and were shown to rupture red blood cells upon capture to release and ingest their contents. In addition, C. elegans survived equally on a diet of erythrocytes. There was no statistically significant difference in survival when compared with a diet of Escherichia coli OP50. The enzymes responsible for the digestion and detoxification of haem and haemoglobin, which are key components of the hookworm vaccine, were found in the C. elegans intestine. These findings support our postulate that free-living nematodes could provide a model for the assessment of neutralizing antibodies to current and future hematophagous parasite vaccine candidates.

Polymerase chain reaction (PCR) based amplification of hmcp3 and hmcp6 cysteine protease genes of Haemonchus contortus from small ruminants.

Haemonchus contortus is a haematophagous nematode in small ruminants population and causes anaemia, weakness and mortality especially in young animals. In the present study, hmcp3 partial gene of 836bp and hmcp6 full length gene of 1041bp were amplified from the cDNA of Bareilly isolate of adult male H. contortus by polymerase chain reaction. Further gene characterization and expression studies are warranted to know the immunoprophylactic potential of hmcp3 and hmcp6 proteins of H. contortus.

Epidemiology of gastrointestinal nematodosis in goats maintained at farm and field conditions in chhattisgarh plain

Studies were conducted to understand nematode parasite epidemiology in goats maintained under field and farm condition at Durg situated in the agroclimatic zone of Chhattisgarh Plain by monitoring monthly faecal egg counts and genera of parasites present for a period of one year. While increased egg counts in goats were observed during July to September in goats of both field and farm conditions, the egg counts were significantly (P<0.05) higher in field goats with dominance of haematophagous nematode Haemonchus spp.

Biased Sex Ratio in the European Eel (Anguilla anguilla) Swim-Bladder Parasite Anguillicola crassus, Experimentally Induced by 11-Ketotestosterone

Parasites are intimately connected to the host in which they live, and some may be affected by the polluted environment of their host. The present study describes the effect of a steroid hormone (11-ketotestosterone) on the sex ratio of the invasive hematophagous nematode Anguillicola crassus Kuwahara, Niimi & Itagaki, 1974, when experimentally injected to European eels, Anguilla anguilla. Our results showed that this steroid induced a significant male-biased ratio in the nematode A. crassus infrapopulations, suggesting that the presence of endocrine disruptors in the environment may lead to skewed sex ratios among parasites.

Intermolecular Interactions and Characterization of the Novel Factor Xa Exosite Involved in Macromolecular Recognition and Inhibition: Crystal Structure of Human Gla-domainless Factor Xa Complexed with the Anticoagulant Protein NAPc2 from the Hematophagous Nematode Ancylostoma caninum

NAPc2, an anticoagulant protein from the hematophagous nematode Ancylostoma caninum evaluated in phase-II/IIa clinical trials, inhibits the extrinsic blood coagulation pathway by a two step mechanism, initially interacting with the hitherto uncharacterized factor Xa exosite involved in macromolecular recognition and subsequently inhibiting factor VIIa ( K i = 8.4 pM) of the factor VIIa/tissue factor complex. NAPc2 is highly flexible, becoming partially ordered and undergoing significant structural changes in the C terminus upon binding to the factor Xa exosite. In the crystal structure of the ternary factor Xa/NAPc2/selectide complex, the binding interface consists of an intermolecular antiparallel β-sheet formed by the segment of the polypeptide chain consisting of residues 74–80 of NAPc2 with the residues 86–93 of factor Xa that is additional maintained by contacts between the short helical segment (residues 67–73) and a turn (residues 26–29) of NAPc2 with the short C-terminal helix of factor Xa (residues 233–243). This exosite is physiologically highly relevant for the recognition and inhibition of factor X/Xa by macromolecular substrates and provides a structural motif for the development of a new class of inhibitors for the treatment of deep vein thrombosis and angioplasty.

Hookworms hit the headlines in the New Millennium

The year 2000 was a good year for hookworm researchers and possibly a bad year for hookworms. After many years in the parasitological doldrums, these haematophagous nematode parasites of humans (Necator americanus and Ancylostoma duodenale) have finally attracted some serious attention from the funding organizations, reflecting their importance in causing disease. A major award from the Bill and Melinda Gates Foundation to the Sabin Vaccine Institute and the Dept of Microbiology and Tropical Medicine at George Washington University will underpin the development of a hookworm vaccine, and the inclusion of hookworms in the Wellcome Trust-funded Beowulf Initiative will significantly enhance our knowledge of the molecular genetics and fundamental biology of these interesting parasites. Does this signal the end of the humble hookworm? Probably not in the short-term. Hookworms are adept immune regulators, and have the capacity to downregulate natural and possibly vaccine-induced immunity. Furthermore, there might be immunological situations where hookworm infections of tolerable intensity could be beneficial. DP

Inherent flexibility in a potent inhibitor of blood coagulation, recombinant nematode anticoagulant protein c2.

Nematode anticoagulant proteins (NAPs) from the hematophagous nematode Ancylostoma caninum inhibit blood coagulation with picomolar inhibition constants, and have been targeted as novel pharmaceutical agents. NAP5 and NAP6 inhibit factor Xa by binding to its active site, whereas NAPc2 binds to factor Xa at a different, as yet unidentified, site and the resultant binary complex inhibits the tissue factor-factor VIIa complex. We have undertaken NMR studies of NAPc2, including the calculation of a solution structure, and found that the protein is folded, with five disulfide bonds, but is extremely flexible, especially in the acidic loop. The Halpha secondary shifts and 3JHNHalpha coupling constants indicate the presence of some beta structure and a short helix, but the intervening loops are highly conformationally heterogeneous. Heteronuclear NOE measurements showed the presence of large amplitude motions on a subnanosecond timescale at the N-terminus and C-terminus and in the substrate-binding loop, indicating that the conformational heterogeneity observed in the NMR structures is due to flexibility of the polypeptide chain in these regions. Flexibility may well be an important factor in the physiological function of NAPc2, because it must interact with other proteins in the inhibition of blood coagulation. We suggest that this inhibitor is likely to become structured on binding to factor Xa, because the inhibition of the tissue factor-factor VIIa complex requires both NAPc2 and factor Xa.

Synthesis and biological activity of a series of diaryl-substituted .alpha.-cyano-.beta.-hydroxypropenamides, a new class of anthelmintic agents

A series of alpha-cyano-beta-hydroxypropenamides was prepared and tested for anthelmintic activity. alpha-Cyano-beta-hydroxy-N-[4- (trifluoromethyl)phenyl]-3-[4-(trifluoromethyl)phenyl]propenamide (1) showed good activity against the nematode Nematospirodes dubius in a mixed parasite infection in mice; several of the analogues were also effective against the cestode Hymenolepis nana. In sheep trials, 1 caused 100% reduction of the hematophagous nematode Haemonchus contortus after a single dose of 20 mg/kg but did not show satisfactory control of Trichostrongylus colubriformis or Ostertagia circumcincta. Against the liver fluke Fasciola hepatica, 1 suppressed egg production but only temporarily, suggesting that the adult flukes were not eliminated. Mechanism of action studies on 1 using Ascaris mitochondria showed it to be an uncoupler of oxidative phosphorylation.