Worm Surface Research Articles

Synthetic peptide-targeted selection of phage display mimotopes highlights immunogenic features of α-helical vs non-helical epitopes of Taenia solium paramyosin: Implications for parasite- and host-protective roles of the protein

Paramyosin of the pig-human parasite Taenia solium (TPmy) is a α-helical protein located on the worm surface that is suggested to fulfill an immunomodulatory role protecting the parasite against host immune system. Besides, in challenging experiments the protein shows a vaccine potential. These observations imply that TPmy harbors antigenic determinants for each of these contrasting actions. However the suggestion was not given a support from experimental data because respective epitopes have not been described thus far. To circumvent this difficulty, we use synthetic peptides with sequences of regions composed of α-helical or linear structure to induce rabbit antibody responses for phage-display mapping of epitope core amino-acid sets. Antibodies to α-helical regions were weak binders and M13 phage-displayed peptides selected by them from two different libraries exhibited no amino-acid similarities with the original protein site. In contrast, the antibodies produced in response to non-helical segment within α-helical structure were better binders and selectors of perfect structural mimics of the protein site. This first phage display epitope analysis of TPmy supports the notion that the rod-like α-helix, which encompasses over 90% of the total amino acids, may serve as an immunomodulatory shield that protects the parasite. Further, the seven non-helical segments of the TPmy molecule may represent the only anti-parasite discrete immunogenic epitopes whose representative mimotopes can be utilized in development of pure epitope vaccines.

Further studies on mefloquine and praziquantel alone or interaction of both drugs against Schistosoma japonicum in vitro

The aim of the present study is to further understand and analyze the interaction of mefloquine with praziquantel against adult Schistosoma japonicum in vitro. Mice infected with S. japonicum cercariae for 35-37 days were sacrificed, and adult schistosomes were collected by perfusion. Schistosomes were placed to each of 12 wells of a Falcon plate and maintained in RPMI 1640 supplemented by 10% calf serum. For determination of 50% and 95% lethal concentration (LC50 and LC95) of the two drugs in vitro, schistosomes were exposed to mefloquine at concentrations of 1, 2, 3, 4, 5, 6, 7, and 10 μg/mL or praziquantel at concentrations of 0.001, 0.01, 0.05, 0.1, 0.2, 0.5, 1, 10, and 30 μg/mL. The plate was incubated at 37°C in 95% air + 5% CO₂ for 72 h. According to the half-life of oral mefloquine and praziquantel in mice, mefloquine combined with praziquantel simultaneously, mefloquine administered within 1 h after praziquantel and praziquantel administered within 17 h after mefloquine were used to evaluate the effect of mefloquine in combination with praziquantel against S. japonicum in vitro. The results showed that the LC50 and LC95 of mefloquine calculated by the Bliss method were 6.17 μg/mL (95% confidence limits, 5.84-6.517 μg/mL) and 8.703 μg/mL (95% confidence limits, 7.632-9.797 μg/mL), respectively. As to praziquantel, no worm death was seen when schistosomes were exposed to praziquantel at concentrations of 0.005-0.2 μg/mL for 72 h. While in the worms exposed to praziquantel 1, 10, and 30 μg/mL, strong spasmodic contractions of the worm body and vesiculation along the worm surface were observed, but 48-75% of the schistosomes survived the exposure in 72-h incubation. Meanwhile, the number of dead worms that emerged in each group was not proportion to the increasing concentrations. Therefore, it is not appropriate to calculate the LC50 and LC95 of praziquantel. For evaluation of the interaction with the two drugs, praziquantel 0.1 or 0.2 μg/mL, which may induce moderate or strong spasmodic contractions of the worm body and vesiculation along the worm surface, was combined with mefloquine 5, 6, or 7 μg/mL. It was found that when mefloquine combined with praziquantel simultaneously or administered 1 h after addition of praziquantel, the spasmodic contraction of the male worm body was antagonized by mefloquine in various degrees according to the concentrations of mefloquine used. Meanwhile, praziquantel-induced weakened motor activity could be reversed by mefloquine. In female worms, morphological alterations and stimulated motor activity induced by mefloquine still developed. Interestingly, using these two regimens to combine mefloquine with praziquantel resulted in no impact or a decrease in worm mortality. On the other hand, praziquantel 0.2 μg/mL administered within 17 h after mefloquine 5 or 6 μg/mL promoted the damage to the tegument of the worms, which led to enhance the worm mortality compared with that of worms exposed to mefloquine alone. The results indicate that in vitro higher concentrations of praziquantel administered within 17 h after mefloquine may increase the effect against adult schistosomes, while praziquantel combined with mefloquine simultaneously or administered 1 h before addition of mefloquine exhibits no impact or decrease in the effect against schistosomes.

Cross-reaction of antigen preparations from adult and larval stages of the parasite Setaria equina with sera from infected humans with Wuchereria bancrofti

Crude antigenic preparations from Setaria equina were used in ELISA and Western blotting to examine cross-reaction with human sera from areas endemic for bancroftian filariasis. Sera from normal subjects from non-endemic areas were included as negative controls. Cross-reaction was found between S. equina antigens and antibodies in the sera of Wuchereria bancrofti-infected patients, with the highest levels observed between sera of chronic infected patients and Setaria spp. crude female worm surface antigen (CFSWA). In the absence of active transmission of Setaria spp. infection, CFWSA is useful to detect chronic W. bancrofti infection before patients become symptomatic, particularly when chronic patients are known to be amicrofilaraemic. In the presence of active S. equina infection, antigens from the adult and microfilaraemic stages showed the highest degree of cross-reaction with human sera.

Cloning, expression, and characterization of Schistosoma japonicum tegument protein phosphodiesterase-5

The tegument proteins of schistosomes are regarded as potential vaccine candidates and drug targets to control schistosomiasis. Nucleotide pyrophosphatase/phosphodiesterase-5 (NPP-5), which belongs to a multigene family of nucleotide pyrophosphatase/phosphodiesterases (NPPs), is important in the hydrolysis of pyrophosphate or phosphodiester bonds in nucleotides and their derivatives. In the present study, SjNPP-5, identified as one of the tegument proteins of Schistosoma japonicum in our previous proteomic studies, was cloned on a fragment of 1,371 bp and expressed as a recombinant protein of 69 kDa. Real-time RT-PCR analysis showed that SjNPP-5 was up-regulated at 21-42 days, and the expression level in 42-day-old male worms was almost nine times higher than that in females. Western blot analysis revealed that rSjNPP-5 had good antigenicity. Immunofluorescence analysis found that SjNPP-5 was a membrane-associated antigen mainly distributed on the surface of the male adult worm of S. japonicum. BALB/c mice vaccinated with rSjNPP-5 three times showed a 29.90% worm reduction (P < 0.05) and a 26.21% egg count reduction (P > 0.05). Immunization with rSjNPP-5 induced a mixed Th1/Th2 response in which Th1 was dominant. The response was characterized by a reduced IgG1/IgG2a ratio and elevated production of cytokines IFN-γ and IL-4. This study suggested that SjNPP-5 may be important in schistosome development, and further investigations are required to fully understand the function of this molecule.

Ultrastructural alterations of juvenile Schistosoma japonicum harbored in mice following mefloquine administration

The aim of the present study was to assess the ultrastructural alterations of juvenile Schistosoma japonicum induced by mefloquine. Mice infected with 14-day-old S. japonicum were treated orally with mefloquine at a single dose of 400 mg/kg. Between 8 h and 7 days after treatment, groups of two mice were sacrificed, and schistosomula were recovered for transmission electron microscopic observations. Ultrastructural damage was seen in the tegument, subtegumental musculature, parenchymal tissues, and gut epithelial cell. It was already prominent 8 h after drug administration and increased in severity rapidly to reach a peak 3 days post-treatment. Tegumental alterations were characterized by emergence of irregular and elongated cytoplasmic processes, which further fused together accompanied by indistinction of matrix and roughness of external plasma membrane. Meanwhile, in the subtegument, damage to the syncytium, swelling, and lysis of muscle bundles and parenchymal tissues were universal, which further aggravated the lesion on the tegument, followed by collapse or disintegration of damaged tegument to form numerous fragment or debris of cytoplasmic process detached from the worm surface. Severe damage to the gut epithelial cell was also observed 8 h post-mefloquine treatment, which included focal lysis of cytoplasm accompanied by formation of vacuoles and degeneration of mitochondria, emergence of enlarged and contracted nucleus with indistinct or focal disrupted nuclear membrane, and decrease in microvilli. All these alterations further increased in severity and reached the peak 3 days post-treatment. The findings of our study indicate that mefloquine exhibits a fast and potent ability to cause extensive ultrastructural damage to juvenile S. japonicum, which correlates with its high efficacy against juvenile schistosomes.

Tribological Properties of Fe-Cu-MoS&lt;sub&gt;2&lt;/sub&gt; and Self-Lubricating Behaviors

The Fe-Cu-MoS2composites with different adding amount of MoS2as lubricant were prepared by induction sintering method. Their mechanical and tribological properties from room temperature (RT) to 800°C were tested by universal testing machine and high temperature tribometer. The effects of amounts of MoS2, temperature, load and sliding distance on the friction and wear properties of composite were discussed. The structure of the composite was analyzed by XRD and worm surface morphologies were observed by SEM. It was found that MoS2was decomposed during the hot-press sintering process. Meanwhile, solid solution alloy of Mo and Fe, and sulfides were formed in composite, which were responsible for low-friction and high wear-resistance at elevated temperature, respectively. Hardness and anti-compress strength can be improved by adding 8 wt. % MoS2. The friction coefficients and wear rates of composites decrease with the increase of adding amount of MoS2until a critical value of 8 wt. %. The composite with 8 wt. % MoS2shows the optimum tribological properties over the temperature range of RT~800°C.

Characterization and expression of a novel cystatin gene from Schistosoma japonicum

Cystatins are a family of cysteine protease inhibitors that play a crucial role in the immune evasion from their host and in the adaptation to host defence. Here, we isolated a full-length cDNA sequence inferred to encode a novel cystatin gene from a blood fluke, Schistosoma japonicum. The cDNA, designated SjCystatin, comprised an open reading frame (ORF) of 306 bp, and encoded 101 amino acids with a predicted molecular weight of 11.3 kDa. This predicted protein shared a significant degree of sequence identity with the type I cystatin (stefin) of Schistosoma mansoni and Homo sapiens. These proteins exhibited a typical cystatin topology, including the absence of disulfide bonds and three conserved catalytic motifs, Gly at the N-terminus (Gly 6), Gln-X-Val-X-Gly motif (Q 49VVAG 53) and an LP pair at the C-terminus (L 76P 77). The SjCystatin gene spanned 376 bp and contained three exons. The positions of two introns were conserved between the cystatin genes of trematodes and their vertebrate hosts. Reverse transcription polymerase chain reaction confirmed the transcription of SjCystatin in the egg, schistosomula and adult stages of S. japonicum. The encoding ORF region was cloned into pET-28a (+) prokaryotic expression vector. After purification, the recombinant protein SjCystatin (recSjCystatin), expressed in Escherichia coli, was used to immunize animals and produce its specific polyclonal antibody. Western blot analysis revealed that the native SjCystatin was expressed in the egg and adult stages. The enzyme activity assay of the recSjCystatin showed that it inhibited the proteolytic activity of papain. SjCystatin protein was mainly localized on the miracidium within eggs. Immunohistochemistry revealed that SjCystatin mainly localized in the epithelial cells lining the gut as well as the tegument on the surface of adult worms. The conserved genomic DNA structure among cystatin homologues of trematode and their vertebrate host emphasized the characteristics of compatibility between parasites and their hosts. This study provides the first insight into the gene and protein of S. japonicum cystatin and a basis for a further understanding the functions of this gene.

Enzymatic activity and immunolocalization of Schistosoma mansoni and Schistosoma haematobium neutral sphingomyelinase

We predicted, and provided evidence for, the existence of a schistosome tegument-associated Mg 2+-dependent neutral sphingomyelinase (nSMase), which controls hydrolysis of surface membrane sphingomyelin molecules, thus allowing nutrients, but not host antibodies, to access proteins at the host–parasite interface. While a putative nSMase was identified in a recent Schistosoma mansoni genome sequencing and analysis study, our report is the first to measure nSMase enzymatic activity in Triton X-100-solubilized surface membrane (Sup 1) and whole worm soluble (SWAP) molecules of male and female S. mansoni and Schistosoma haematobium. Neutral, but no acidic, sphingomyelinase activity was readily detectable by the amplex red sphingomyelinase assay, and increased with incubation time and protein amount. Like nSMase family members, the schistosome nSMase activity was significantly ( P < 0.05 to <0.0001) enhanced by unsaturated fatty acids and phosphatidyl serine and significantly ( P < 0.01) decreased following exposure to the nSMase specific inhibitor GW4869. Peptides based on the published sequence of S. mansoni putative nSMase and used in a multiple antigen peptide form induced the generation of specific antibodies, which readily bound to the immunogen and to the cognate protein in Sup 1 and SWAP. Immunofluorescence studies suggested the parasite nSMase is located in the worm tegument and gut lining. Studies using RNA interference are in progress to define nSMase role in larval and adult worm surface membrane antigen exposure and unsaturated fatty acid-mediated attrition.

Schistosomicidal and antifecundity effects of oral treatment of synthetic endoperoxide N-89

1,2,6,7-Tetraoxaspiro[7.11]nonadecane (N-89) is a chemically synthesized compound with good efficacy against malaria parasites. We observed strong anti-schistosomal activities of N-89 both in vitro and in vivo. In a murine model with experimental infection of Schistosoma mansoni, orally administered N-89 at the dose of 300 mg/kg resulted in a significant reduction in worm burden (63%) when mice were treated at 2-weeks postinfection. Strong larvicidal effects of N-89 were confirmed in vitro; schistosomula of S. mansoni were killed by N-89 at an EC50 of 16 nM. In contrast, no significant reduction in worm burden was observed when N-89 was administered at 5 weeks postinfection in vivo. However, egg production was markedly suppressed by N-89 treatment at that time point. On microscopic observation, the intestine of N-89-treated female worms seemed to be empty compared with the control group, and the mean body length was significantly shorter than that of controls. Nutritional impairment in the parasite due to N-89 treatment was possible, and therefore quantification of hemozoin was compared between parasites with or without N-89 treatment. We found that the hemozoin content was significantly reduced in N-89 treated parasites compared with controls ( P < 0.001). The surface of adult worms was observed by scanning and transmission electron microscopy, but there were no apparent changes. Taken together, these observations suggested that N-89 has strong antischistosomal effects, probably through a unique mode of drug efficacy. As N-89 is less toxic to mammalian host animals, it is a possible drug candidate against schistosomiasis.

Is the establishment rate and fecundity of Haemonchus contortus related to body or abomasal measurements in sheep?

The relationship among parasitological parameters, abomasal size and body size measurements was investigated in lambs following an experimental infection with Haemonchus contortus. In total, 100 lambs from five different genotypes (German Merino (GM), Texel × GM, Suffolk × GM, German Blackhead Mutton × GM and Ile de France × GM) were experimentally infected with 5000 infective third stage larvae of H. contortus at the time of weaning at 12 weeks of age. Four and six weeks after infection, individual faecal samples were collected for estimation of faecal egg counts (FECs). Furthermore, wither height, shoulder width, heart girth, loin girth and body length were taken at 18 weeks of life. Lambs were slaughtered and necropsied 7 weeks post-infection, and worm counts, abomasal volume and surface area were determined. Positive correlations were found between different body size parameters, body weight and abomasal sizes. FEC and worm counts were not significantly correlated either with body size parameters or with abomasal size. The mean worm burden was higher in GM than in crossbred lambs. There was no significant difference in abomasal size between GM and crossbred lambs. The results suggest that the variations between animals in worm burden following an experimental infection with H. contortus (worm resistance) are not influenced by body size parameters or abomasal sizes. Therefore, other factors, including genetic-based differences in resistance, must cause these findings between and within breeds.

Identification and characterization of a serine protease inhibitor with two trypsin inhibitor-like domains from the human hookworm Ancylostoma duodenale

Protease inhibitors play important roles in the parasitic nematodes' survival within their host, in the development and reproduction of the parasites. The present study described the isolation, identification, and characterization of a novel member of the Ascaris family of serine protease inhibitors, designated AduTIL-1, from the human hookworm Ancylostoma duodenale. AduTIL-1 is composed of a signal sequence and two trypsin inhibitor-like (TIL) domains, which showed the highest similarity with OdmCRP, a putative serine protease inhibitor with two TIL domains in Oesophagostomum dentatum. Each TIL domain of the AduTIL-1 was expressed in Escherichia coli, and their inhibitory activities against serine proteases from animals and human were characterized, respectively. Both of the two TIL domains inhibited human neutrophil elastase and pancreatic trypsin, but different in effectiveness. Although the first TIL domain of AduTIL-1 inhibited bovine pancreatic chymotrypsin (Ki=18.0 nM), both of the two domains showed no inhibitory activity against the human pancreatic chymotrypsin. Immunohistochemical studies demonstrated that AduTIL-1 was localized in esophagus, intestine, and cuticular surface of the adult worms. These results suggested that AduTIL-1 may be involved in the survival of A. duodenale in host by targeting related digestive enzymes and neutrophil elastase.

Schistosoma mansoni host-exposed surface antigens characterized by sera and recombinant antibodies from schistosomiasis-resistant rats

Antibodies from Schistosoma mansoni-infected rats, unlike mice, show a higher titer for schistosome apical tegumental antigens compared with non-apical membrane antigens. These antibodies bind to the surface of living lung-stage worms and to formaldehyde-fixed adult worms. We produced a single-chain antibody Fv domain (scFv) phage library displaying the antibody repertoire of rats highly immune to schistosome infection and we selected for scFvs that recognize the host-exposed surface of worms. Five unique rat scFvs (Teg1, Teg4, Teg5, Teg20 and Teg37) were obtained which recognize schistosome surface epitopes. Each of the scFvs recognizes the surface of living schistosomula and lung-stage schistosomules and/or the surface of formaldehyde-fixed adult worms. None of these scFvs reproducibly stained living adult worms. This suggests that a change occurs during the transition from lung schistosomules to 4-week adults such that at least some surface antigens, although remaining on the surface in living adult worms, can no longer be immunologically stained. Teg1 and Teg4 scFvs both recognize specific bands on Western blots. No bands were observed for the other three scFvs, suggesting that these scFvs may recognize non-protein or conformationally-dependent epitopes. Teg1 was unambiguously identified as recognizing the S. mansoni tetraspanin antigen, SmTSP-2, within the large extracellular domain. Teg4 recognizes a 35 kDa band tentatively identified as Sm29 by proteomic analysis. These scFvs can now be used to characterize schistosome epitopes at the host-parasite interface, to target worms in vivo, and to study the mechanisms by which these worms naturally evade immune damage to the tegument within permissive hosts.

Cloning and characterization of a plasminogen-binding surface-associated enolase from Schistosoma bovis

Schistosoma bovis is a ruminant parasite able to survive prolonged periods in the vasculature of its host without either being cleared by the host defensive systems or inducing thrombotic or coagulation disturbances. This suggests that the parasite modulates both the immune and haemostatic host responses. Previous studies have shown that host plasminogen binds to the surface of S. bovis adult worms, and that a tegument extract from S. bovis fixes and activates host plasminogen, generating plasmin, which in turn could both inhibit blood clotting and dissolve clots. Enolase has been identified among the tegumental proteins that bind plasminogen. The aim of the present study is to determine the physiological role of the enolase found in the tegument of S. bovis adult worms as regards plasminogen-binding and activation, and to confirm its surface exposure on the parasite. The study included the cloning and sequencing of S. bovis enolase cDNA, collection of the corresponding recombinant protein and evaluation of its plasminogen-binding and activation activity, and an exploration of the expression and localization of native enolase in adult worms and lung schistosomulae. Here we show that S. bovis male adult worms express enolase on their tegumental surface and that this protein binds host plasminogen and increases its activation in the presence of host tissue plasminogen activator (t-PA). This suggests that the surface-associated enolase found here is a physiological receptor of plasminogen that plays a role in the activation of the host fibrinolytic system, most probably to avoid blood clot formation on the worm's surface.

The Conjugation of the Hardened and Grinded Worm Gear

Research of the new kind of meshing is an important research direction of mechanics and manufacture. At present, we have only few methods. Such as the Oliver first method, the Oliver second method and the F. Litven’s method. This paper presents a new kind of conjugate method. The new method is of great significant both in theory and in practice. In order to save nonferrous metal and to increase the strength of the worm gear, scholars at home and abroad suggested producing the worm gear with steel and iron. In other words, both the surface of worm and worm gear are hardened and grinded. So that it is necessary to probe the new way of conjugation.

A Preliminary Study of the Surface Properties of Earthworms and Their Relations to Non-Stain Behaviour

The surface properties of earthworms were studied using nitrogen adsorption/desorption isotherm and dynamic contact angle measurement with the aim to understand their non-stain behaviour. The results obtained by applying dynamic contact angle technique using water, glycerol, cooking oil and dimethylsilicone show that the surface properties of earthworms are a function of time. The critical surface energy, calculated using advancing angle, is as low as 11 × 10 −3 J·m −2. However this hydrophobic behaviour at the initial contact moment changes progressively into hydrophilic as time goes by. This behaviour together with the creeping movement of corrugated surface is believed to be responsible for the non-stain behaviour of earthworms. The nitrogen adsorption isotherm of dried skin of earthworms at 77.3 K exhibits more or less Type V isotherm with surface area of 13 m 2·g −1 calculated using the α s plot. The Type V isotherm is the indication of weak interaction between nitrogen and the worm surface.

The Mannose-Binding Protein Sm60 from Schistosoma mansoni Suppresses T Cell Proliferation via Inhibition of Interleukin-2 Production

Adult worms of Schistosoma mansoni live in the bloodstream, employing immune evasion strategies and exposing few antigens to the immune system. The tegumental surface of adult worms presents Sm60, a mannose-binding protein, which induces neutrophil migration and mast cell degranulation. Here we demonstrated that Sm60 is able to block the in vitro antigen-specific or polyclonal spleen cell proliferation induced by keyhole limpet haemocyanin (KLH) or con- canavalin A (Con A), respectively. To address the mechanism of inhibition, we evaluated some cytokines in culture and observed that Sm60 decreased significantly the synthesis of interleukin (IL)-2 induced in response to KLH, but not other cytokines. To block the suppression triggered by Sm60, exogenous IL-2 or α-methyl-mannoside were added to KLH- stimulated cultures pulsed with Sm60. Only IL-2 abolished the Sm60 effect, suggesting that Sm60 blocked the lymphop- roliferation through the inhibition of IL-2 production, in a carbohydrate recognition domain (CRD)-independent mecha- nism. Our results suggest that Sm60 could participate in the immune evasion mechanisms of S. mansoni.

Theory and Application of Surface Generated by Curvilinear Cutter Edge with Two Degrees of Freedom

In order to study the mesh problem in case that the surface is generated by curvilinear cutter with two degrees of freedom a theory of surface generated by curvilinear cutter edge with two degrees of freedom is put forward by using the conjugate conditions of two degrees of freedom motion. The theory is used to calculate the helical surface of the toroidal worm generated by linear cutter edge whose relative motion has two degrees of freedom. This kind of worm is manufactured on a hob machine and its helical surface of the worm is measured on a CMM. The application prospect of this kind of worm gears is studied. The conclusions are: by using the theory the generate curved surface can be deduced when the curvilinear cutter edge and the varying regularity of the two motion parameters are given or the varying regularity of the two motion parameters can be derived when the generated curved surface and the curvilinear cutter edge are given;the calculation of the curved surface generated by curvilinear cutter edge with two degrees of freedom can be simplified if the varying regularity of the two motion parameters is restricted properly; the "modification" curve of the worm generated by linear cutter edge with two degrees of freedom is close to that of the hindley worm which is modified in parabolic curve; the defect of the hindley worm gears can be avoided by using this kind of worm gears without needing modification and there is a good application prospect.

A mathematical model of a worm mechanism generated by a conical cutter

In this article, an imaginary conical cutter was used to determine the geometric models of the worm—gear mechanism including a worm and a gear. Mathematical models of the worm and the gear were derived using an imaginary conical cutter and gear theory. Based on the geometric relations between the worm and conical cutter surfaces and between the gear and conical cutter surfaces, the worm and the gear were regarded as an envelope to the family of conical cutter surfaces when they rotated for one cycle. Using the developed mathematical model, the stress analysis of the proposed mechanism was examined. The results show that the von Mises stress and volume of the proposed worm and gear were smaller than those of the conventional worm—gear mechanism. Lastly, a numerical example was used to illustrate the geometric model of a gear and a worm with a gear ratio of 30:1.

Intraspecific Epitopic Variation in a Carbohydrate Antigen Exposed on the Surface of Trichostrongylus colubriformis Infective L3 Larvae

The carbohydrate larval antigen, CarLA, is present on the exposed surface of all strongylid nematode infective L3 larvae tested, and antibodies against CarLA can promote rapid immune rejection of incoming Trichostrongylus colubriformis larvae in sheep. A library of ovine recombinant single chain Fv (scFv) antibody fragments, displayed on phage, was prepared from B cell mRNA of field-immune sheep. Phage displaying scFvs that bind to the surface of living exsheathed T. colubriformis L3 larvae were identified, and the majority of worm-binding scFvs recognized CarLA. Characterization of greater than 500 worm surface binding phage resulted in the identification of nine different anti-CarLA scFvs that recognized three distinct T. colubriformis CarLA epitopes based on blocking and additive ELISA. All anti-CarLA scFvs were specific to the T. colubriformis species of nematode. Each of the three scFv epitope classes displayed identical Western blot recognition patterns and recognized the exposed surface of living T. colubriformis exsheathed L3 larvae. Surprisingly, each of the anti-CarLA scFvs was able to bind to only a subset of worms. Double-labelling indirect immunofluorescence revealed that the three classes of anti-CarLA scFvs recognize distinct, non-overlapping, T. colubriformis sub-populations. These results demonstrate that individual T. colubriformis L3 larvae display only one of at least three distinct antigenic forms of CarLA on their surface at any given time, and suggest that antigenic variation within CarLA is likely a mechanism of immune evasion in strongylid nematodes.

Checking on the Simulation of the Plane Enveloping Toroidal Worm

The three-dimensional figure of a toroidal worm is obtained according the simulation based on the mathematical model. The key of this is the authenticity of the worm surface expressed. Comparing a simulating worm with the true worm, the conformational degree between them can be found out. As the worm surfaces are complex space surfaces, so it is difficult to determine the measuring method and reference for the surfaces of a worm. According to the entire analysis's, the symmetry section of the worm is defined as a representing section. The space surface measuring is converted to a plane curve measuring, and the possibly existing error will be analyzed. The experiments for checking two true worms are made to prove the truthfulness and reliability of the simulation program of the plane enveloping toroidal worm.