Search

Coenurosis is a serious parasitic disease of herbivorous animals caused by the metacestode of Taenia multiceps (Coenurus cerebralis). Accordingly, a significant amount of research is currently dedicated to the development of appropriate antigens for use in rapid and accurate coenurosis diagnosis kits. In the present study, antigen B (AgB) and thioredoxin peroxidase (TPx) from T. multiceps were cloned and expressed using a prokaryotic system, molecular characterization of Tm-AgB was determined by bioinformatical analyses. The serological diagnostic potentials of rTm-AgB and rTm-TPx were evaluated by indirect ELISA and compared with those of previously reported rTm-AnxB2, rTm-HSP70, and rTm-GST. The results showed that Tm-AgB is a specific lipoprotein of cestodes with good thermal stability. The ELISA assay showed that rTm-AgB exhibited a sensitivity of 95.8% and a specificity of 87.5%, indicating its strong potential for serological diagnosis of T. multiceps.

Alveolar echinococcosis (AE) is a worldwide zoonosis caused by E. multilocularis . Humans become infected through oral ingestion of the eggs. Host of E. multilocularis produces immune responses that help to either reject and/or limit the growth of this parasite, and in response the parasite produces molecules against this immune attack. This study identifies candidate key molecules in the early infection phase and the chronic stage of the parasite infestation, through comparison of gene expression of 4- and 16-week metacestodes. First, RNA was isolated from 4- and 16-weeks metacestodes of E. multilocularis (Nemuro strain). Thereafter, clean reads with lengths of 50 bp or longer were compared against a reference genome using TopHat. Functional annotation of transcripts of E. multilocularis were investigated using multi-step bioinformatics tools. At the gene ontology (GO) level, 356 and 1774 transmembrane (TM) predicted proteins of the E. multilocularis were mapped to an enhanced ‘hydrolase activity’ and increased ‘transmembrane transporter activity’, respectively. In addition, comparison of gene expression level between 4- and 16-week metacestode revealed 168 different expression (DE) genes. This study has demonstrated that, the expression levels of predicted ES and TM proteins in E. multilocularis change in the transformation from one stage to another. Genes that are highly expressed in immature or mature metacestode could be explored as novel candidates for diagnostic antigens and vaccine targets.

Taenia ovis metacestodes reside in the muscle of sheep and goats, and may cause great economic loss due to condemnation of carcasses if not effectively controlled. Although advances have been made in the control of T. ovis infection, our knowledge of T. ovis biology is limited. Herein the protein profiling of T. ovis metacestodes was determined by liquid chromatography-linked tandem mass spectrometry. A total of 966 proteins were identified and 25.1% (188/748) were annotated to be associated with metabolic pathways. Consistently, GO analysis returned a metabolic process (16.27%) as one of two main biological process terms. Moreover, it was found that 24 proteins, including very low-density lipoprotein receptor, enolase, paramyosin and endophilin B1, were abundant in T. ovis metacestodes. These proteins may be associated with motility, metabolism, signaling, stress, drug resistance and immune responses. Furthermore, comparative analysis of 5 cestodes revealed the presence of Taenia-specific enolases. These data provide clues for better understanding of T. ovis biology, which is informative for effective control of infection.

CC-BY-NC-ND, Two putative serpin genes were identified in Echinococcus multilocularis, in addition to the already reported serpinEmu, and were designated as serpin2Emu and serpin3Emu. Western blot analysis using polyclonal antibodies against serpinEmu, putative serpin2Emu protein, and putative serpin3Emu protein indicated that all three proteins were localized in both intracellular and excretory-secretory (ES) fractions of E. multilocularis metacestodes. In addition, immune staining of parasite tissue indicated that all three proteins were localized at the germinal layer.

Neurocysticercosis (NCC) is a parasitic infection of central nervous system (CNS). Expression of adhesion molecules, chemokines and matrix metalloproteinases (MMPs) were investigated on brain tissues surrounding viable (n=15) and degenerating cysticerci (n=15) of Taenia solium in swine by real-time RT-PCR and ELISA. Gelatin gel zymography was performed for MMPs activity. ICAM-1 (intercellular adhesion molecule-1), E-selectin, MIP-1α (macrophage inflammatory protein-1α), Eotaxin-1 and RANTES (regulated on activation, normal T cell expressed and secreted) were associated with degenerating cysticerci (cysts). However, VCAM-1 (vascular cell adhesion molecule-1), MCP-1 (monocyte chemotactic protein-1), MMP-2 and MMP-9 were associated with both viable and degenerating cysts. In conclusion, viable and degenerating cysticerci have different immune molecule profiles and role of these molecules in disease pathogenesis needs to be investigated.

Cysticercosis caused by the metacestode stage of Taenia hydatigena is endemic in Sardinia. Information on the genetic variation of this parasite is important for epidemiological studies and implementation of control programs. Using two mitochondrial genes, the cytochrome c oxidase subunit 1 (cox1) and the NADH dehydrogenase subunit 1 (ND1) we investigated the genetic variation and population structure of Cysticercus tenuicollis from Sardinian intermediate hosts and compared it to that from other hosts from various geographical regions. The parsimony cox1 network analysis indicated the existence of a common lineage for T. hydatigena and the overall diversity and neutrality indices indicated demographic expansion. Using the cox1 sequences, low pairwise fixation index (Fst) values were recorded for Sardinian, Iranian and Palestinian sheep C. tenuicollis which suggested the absence of genetic differentiation. Using the ND1 sequences, C. tenuicollis from Sardinian sheep appeared to be differentiated from those of goat and pig origin. In addition, goat C. tenuicollis were genetically different from adult T. hydatigena as indicated by the statistically significant Fst value. Our results are consistent with biochemical and morphological studies that suggest the existence of variants of T. hydatigena.

Graphical abstract A: Fattening pigs in a pig barn with outdoor access. B: Echinococcus multilocularis (E.m.) liver lesion in a naturally infected pig. C: histology of a E.m. liver lesion in a naturally infected pig., Highlights • Minimal prevalence for Echinococcus multilocularis infection in Swiss domestic pigs was established. • Minimal prevalence was 0.008% for fattening pigs and 0.11% for breeding pigs, respectively. • All but one metacestode were degenerated; no protoscoleces were detected in histopathology. • Main risk factors were foxes or other animals in the barns, no hygiene barrier, outdoor feeding and grass feeding. • No geographical clusters of higher infection with E. multilocularis were evident., The canid tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE) in humans and other intermediate hosts. Depending on the permissiveness of the intermediate host, the larval form of E. multilocularis (metacestode) may be either fertile, e.g. in rodents, and thus supporting the life cycle of the parasite, or infertile, e.g. in pigs, and thus interrupting the life cycle. Pigs have been shown to act as aberrant hosts for the metacestode and consequently develop liver lesions but represent a dead-end for the parasite. Routine liver inspection at slaughter provided the basis for a large-scale surveillance study on E. multilocularis infection in pigs. The aim of this combined cross-sectional and case-control study was to estimate the minimal prevalence of E. multilocularis in pigs in Switzerland, to find factors associated with infection, and to assess potential regional clusters of infection. During the 12-month-study period, approximately 85% of all pigs slaughtered in Switzerland were assessed. In total, 450 pig livers with macroscopic lesions suggestive of E. multilocularis infection were analysed. Of those, 200 samples were positive by E. multilocularis-PCR. Thus, the overall minimal prevalence detected by molecular means was 0.009% in all slaughter pigs (200 of 2'143'996), 0.008% in finishing pigs (177 of 2'123'542), and 0.11% in breeding pigs (22 of 20'454). Histology revealed the unique presence of a laminated layer in 105 cases, and an additional germinal layer detected in a single case. Protoscoleces could not be observed in any of the lesions. Factors positively associated with infection were "foxes seen in the pig shed", "foxes on premises", "presence of other animals in the shed", "absence of a hygiene barrier", "outdoor feeding", "feeding grass", "lack of rodent control", "not having own dogs on the farm" and "infrequent deworming of sows". Infection was present in all regions sampled and was representative of the important pig rearing areas of Switzerland, without evidence of any obvious geographical cluster. Conclusively, our study provided further evidence of widespread environmental contamination with E. multilocularis eggs in Switzerland. Furthermore, the absence of protoscoleces in any of the lesions supported the concept that pigs act only as a dead-end host and thus do not contribute to the life cycle of the parasite. Factors associated with E. multilocularis infection were in-line with parasite biology, and many can be addressed by increasing hygiene and management standards.

Mesocestoides spp. are zoonotic cestodes of wild and domesticated carnivores. Although the adult stages are relatively harmless intestinal parasites, the metacestode stages (tetrathyridia) can be responsible for life-threatening peritonitis and pleuritis in several species including dogs, cats, non-human primates and probably man. The aim of the present study was to reveal the spatial distribution pattern of Mesocestoides spp. in the most important final hosts, red foxes (Vulpes vulpes), to analyse the relationship of these patterns with landscape and climate by geographical information systems and to evaluate faecal flotation method for the detection of infection in the final host. Fox carcasses, representing 0.5% of the total fox population were randomly selected out of all the foxes of Hungary. The intestinal tract was examined by sedimentation and counting technique. The sensitivity of the flotation method was evaluated by the testing of the faecal samples of 180 foxes infected with Mesocestoides spp. The prevalence of infection was high in foxes (45.8%; 95% CI=41.0-50.6%), and the parasite was detected in all areas of Hungary. The high prevalence of the parasite in foxes suggests that the infection might also be common in outdoor dogs and cats. Mesocestoides infection could not be detected in any of the foxes by flotation method indicating that the sensitivity of the method is less than 0.6%. Therefore, almost all canine and feline infections remain undetected in the veterinary practice. Based on the statistical analysis, the altitude was the only determinant of the spatial distribution of Mesocestoides spp. indicating that infections in carnivores including dogs and cats can be expected mainly in midland regions (150-750 m above sea level). It might be attributed to the altitude-dependent species richness and abundance of the intermediate and final hosts of the parasite.

Hydatid cysts of Echinococcus felidis are described from the hippopotamus (Hippopotamus amphibius) from Mpumalanga Province, South Africa. Among six hippopotami investigated, hepatic hydatids were found in three. The identification was based on mitochondrial and nuclear DNA sequences. In addition, the rostellar hook morphology was analysed. This is the first morphological description of the metacestode of E. felidis, and the first molecularly confirmed report of the intermediate host of E. felidis in South Africa. The definitive host of E. felidis in South Africa is the lion (Panthera leo).

The metacestode stage of the tapeworm, Taenia ovis, causes cystic lesions in the skeletal and cardiac muscle of sheep, which can result in the condemnation of the entire carcass. In recent years, Canadian farms have seen a marked increase in the number of condemnations due to T. ovis. Mathematical transmission models provide a useful tool for predicting parasite transmission and for evaluating the efficacy of potential control options. To date, no model has been developed exclusively for T. ovis. In the work described here, a compartmental, deterministic transmission model was developed to better understand the transmission dynamics of T. ovis on Canadian sheep farms. The model was intended to be practical, and represent the transmission of infection burdens in lambs that result in carcass condemnation, or transmission to canids. All transmission parameters were obtained from the literature or, when unavailable, expert opinion. The model incorporated each stage of the parasite lifecycle using the most probable transmission route on Canadian sheep farms; including definitive host (guard dogs), intermediate host (pastured lambs), and environment. Based on literature, the model performed as expected, and provided a reasonable estimate of parasite prevalence in lambs. In addition, modeling allowed the efficacy of potential control options to be evaluated and compared. Model simulations suggested that infection risk in market lambs could be eliminated through the regular treatment of guardian dogs every fifth week with an appropriate cestocide, or through eliminating carcass consumption by guardian dogs.

Antigen B (AgB) is a major protein produced by the metacestode cyst of Echinococcus granulosus, the causative agent of cystic hydatid disease. E. granulosus AgB is a gene family of at least five gene loci (B1-B5), each one consisting of several minor variants. We used PCR-SSCP followed by DNA sequencing to evaluate sequence variation and polymorphism of AgB1 in 99 isolates which the 43 were from cattle, 25 of sheep and 31 of human. All samples were analyzed with 12S rRNA-PCR for the strain detection and all of were identified as G1-G3 cluster (E. granulosus sensu stricto). The 16 human, 35 cattle and 25 sheep isolates were yielded the 102 bp band by PCR and these samples were tested by SSCP. As results of the SSCP, different band profiles were detected one each of cattle and human isolates while the other 74 isolates showed same band patterns. The DNA sequence analysis was performed for these two isolates and the other selected 4 isolates and polymorphism was confirmed. (C) 2011 Elsevier B.V. All rights reserved.

In the present report metacestodes were collected from the mesentery of a taruca (Hippocamelus antisensis) and from the omentum of a red brocket deer (Mazama americana) in Peru. Various metacestodes parameters, including rostellar hook characteristics, were measured. Molecular analysis was performed to amplify the mitochondrial cytochrome c oxidase subunit 1 gene from metacestode isolates. Metacestodes were identified as T. hydatigena by morphology and molecular methods. This constitutes the first molecular detection of T. hydatigena metacestodes in the taruca and the red brocket deer and demonstrates that these animal species are natural intermediate hosts for this parasite.

Immunodiagnostic confirmation of cystic human hydatidosis is frequently required before surgical intervention or of chemotherapy. However, it remains inadequate to detect specific antibodies or antigens in some confirmed cases of echinococcosis. This study was carried out to investigate the accuracy of three different immunodiagnostic tests for detection of specific circulating antigens or antibodies in the serum and urine of 13 experimentally infected sheep. For this purpose, Echinococcus granulosus were collected from small intestine of experimentally infected dogs, and 2000 taenid eggs were orally administered to each of the 13 sheep. There were six other sheep, which were kept as the control group. Biweekly serum and urine samples were collected from all the sheep for 4 months after infection. The sera were subjected to indirect hemagglutination test and the concentrated urine samples were subjected to coagglutination and counter immunoelectrophoresis tests. The results revealed that the sensitivity of these tests in detecting the hydatid antigens in the urine or antihydatid antibodies in the serum of the infected sheep reached their maximum in 12th and 13th week after infection; then it decreased in the following weeks. Examination of the non-infected sheep samples throughout the experiment showed that the aforesaid findings were specific only to the infected sheep. It seems that the appearance of specific hydatid antigen in urine and its antibodies in the serum were simultaneous. Although these tests are highly specific, false negative outcomes were encountered in their detection of cystic echinococcosis. In general, it seems rational to establish some series of diagnostic procedures in order to reveal antibodies and antigen of metacestode in serum and urine of the patients.

Liver lesions were found in 6/6 pigs 7 months after oral inoculation with 5000 or 35,000 Echinococcus multilocularis eggs. However, lesion morphology differed considerably among the animals. The largest lesions (3-8mm in diameter) were found in a single pig and smaller lesions (1.5-3mm) in three pigs. These lesions were clearly circumscribed and had pronounced central necroses and dystrophic calcifications. In contrast, most of the smallest (usually

Cysticercosis is a zoonotic disease of humans produced by the larval stage of swine parasite, Taenia solium. Chemotherapy of infected pigs is a possible strategy for avoiding disease transmission and improving health programs in endemic areas of cysticercosis. In this preliminary study, seven naturally infected pigs from 6 to 12 months of age were allotted to treated (n = 4) and control groups (n = 3). The treated animals received a subcutaneous injection in their forelegs and thighs of 15 mg/kg per body weight of albendazole sulphoxide (ABZSO; Pisa, Mexico) once per day for 8 days. At the same time, the control group received a subcutaneous injection of saline solution (9% NaCl). After 12 weeks, all the animals were slaughtered and at least 200 metacestodes were isolated from the muscles and brain of each animal. Using histology and the metacestode viability criteria described in this study, treated animals had no viable cysts in their muscle (0/200), while 7 of 17 (41.1%) viable cysts were observed in those isolated from their brains. In the control group, 183/200 (91.5%) muscle metacestodes were viable and from brain, 22/29 (75.8%) metacestodes were viable. The 15 mg/kg per body weight dosage of ABZSO was 100% effective against muscular cysticercosis as shown by the lack of viable cysts and the micro-calcifications in meat from the treated pigs.

Sparganosis is a parasitic infection in amphibians, reptiles and mammals including feral swine and man. It is caused by migration of the metacestode (spargana) of Spirometra. The primary objective of this study was the determination of the prevalence of gross sparganosis in Florida county of origin in slaughtered feral swine. Tracebacks to county of origin were conducted for Florida feral swine with and without gross sparganosis. Feral swine trapped in Florida and presented for slaughter in a Texas slaughter establishment from May to December 1999 was the sample population. Overall prevalence of sparganosis in Florida feral swine was 6.9%. Because Highlands county had the same prevalence, other counties were compared to it. Sparganosis was detected in 17 Florida counties. Swine originating from Osceola or Hillsborough counties (4.3 and 1.8% prevalence, respectively) had lower prevalence of sparganosis than in Highlands, whereas those from Marion county (21.7% prevalence) had a higher prevalence. Transmission to humans may occur via consumption of infected feral swine, other species of secondary intermediate hosts or the primary intermediate hosts.

The aim of the present study was to evaluate diagnostic procedures for porcine cysticercosis. Sera were obtained from 32 pigs reared in commercial farms, 47 pigs before and after experimental infection, 42 carefully necropsied rural pigs and 191 slaughtered pigs from rural communities in which the presence of the Taenia solium metacestode was assessed by tongue dissection. Sera were analyzed by ELISA to detect antibodies against T. solium antigens and to detect parasite antigens. Most sera from the necropsied rural pigs were also evaluated by the Western blot method. Antigen and antibody ELISA detection assays showed high sensitivity and specificity when applied to sera from pigs reared in commercial farms. In contrast, all methods (Ag-ELISA, Ab-ELISA assays, EITB and tongue inspection) showed lower sensitivity and specificity when applied to the generally lightly infected rurally reared pigs. The probability distribution of cysts in carcasses were also determined. These results emphasize the difficulties in detecting cysticercosis in rural pigs with low levels of cyst burdens.

The study was carried out during a 3-year period. Of 9501 cattle examined, 736 (7.7%) were infected with cysticerci of Taenia saginata. The endemic foci were identified and prevalence was significantly higher (P < 0.005) in animals from Noorabad Mammasani (10.7%), Kenareh (10.0%) and Shiraz area (8.5%) than elsewhere. The prevalence was significantly higher (P < 0.005) during spring and autumn seasons. There was no variation in the infection rate in animals of different age groups, suggesting that immunity was acquired to super-infection. The most common sites were muscle of the shoulder (26.3%) and masseter (23.7%), tongue (24.9%) and heart (23.4%). Pharynx, oesophagus and diaphragm showed 0.9, 0.5 and 0.4% infection, respectively. The metacestode caused extensive damage resulting in infiltrative, degenerative changes, haemorrhages, necrosis and exudation mainly in the vicinity of cysts. The infection was the cause of condemnation of 34.6% of infected carcases. The rejected carcases and infected organs were valued at 100.1 million Rials over the 3-year period.

An enzyme-linked immunoelectrotransfer blot (EITB) was developed using soluble Taenia solium metacestode antigen preparation for immunodiagnosis of porcine cysticercosis and compared with an enzyme-linked immunosorbent assay (ELISA). Sera from 20 pigs with parasitologically confirmed cysticercosis were tested by both EITB and ELISA assays. Specificity of the tests was examined by testing 25 serum samples from the UK, where cysticercosis is not endemic and sera from pigs with Echinococcus granulosus (15), Fasciolopsis buski (six), or Trichinella spiralis (five) infections. All but two of the samples from cases of confirmed cysticercosis were positive by EITB and none of the samples from healthy controls or heterologous infections reacted to any of the diagnostic bands. Thus, the test was 90% sensitive and 100% specific. The sensitivity of the ELISA was 70% with 73% specificity, cross-reactions occurring with sera from E. granulosus infected pigs. Four polypeptides (8, 11, 16 and 23 kDa) were identified by SDS-PAGE and EITB that were specifically recognized by pigs with confirmed cysticercosis.

This study was conducted to determine the prevalence and molecular characteristics of Coenurus cerebralis, the metacestode of Taenia multiceps in cattle from Erzurum province located in eastern region of Turkey between November 2009 and April 2010. Five of 1045 brains of cattle (0.47%) were found to be infected with Coenurus cysts. The characteristics and morphology of C. cerebralis were seen in all the cysts. The cyts from three infected cattle were genetically analyzed and confirmed to be T. multiceps metacestodes by NAD1 and COX1 mitochondrial gene sequence analysis. Pairwise comparison between the NAD1 sequences of the T. multiceps isolates from Erzurum and other T. multiceps isolates available in GenBank showed differences ranging from 0.6 to 2.9%, while COX1 sequences showed differences ranging from 0.2 to 2.6%. Considering the two genes, it was seen that all of the three isolates from Erzurum province were in the same group according to phylogenetic analyses. The present findings could provide a stimulus for future studies on the systematic relationships and epidemiology of lesser-known taeniid cestodes in the region, employing mitochondrial sequence data sets. (C) 2010 Elsevier B.V. All rights reserved.

Canine alveolar echinococcosis (AE) is a rare liver disease caused by the larval stage (metacestode) of E. multilocularis . Its diagnosis so far is rather difficult and invasive. Imaging techniques for the diagnosis have been described recently and in this study, the serological diagnosis of canine AE was evaluated in 30 dogs with confirmed AE using ELISA based on seven different antigens ( E. multilocularis -antigens: Em2G11, EmII/3–10, protoscolex (EmP), excretory/secretory and adult integument (EmAd/I); E. granulosus antigens: hydatid fluid (EgHF) and antigen B (EgAgB)) and by the detection of circulating Em2G11-antigen. EmII/3–10 and Em2G11-based tests detected AE in 50 and 53%. The combined serological investigation for specific circulating Em2G11-antigen and specific antibodies against this antigen showed a sensitivity of 77%. This strategy is useful in animals with large parasite masses and ascites. The highest sensitivities for AE were obtained by the EmAd/I and the EmP antigens (97 and 93% sensitivity) with high specificities in the control group of 76 dogs (100 and 98.7%, respectively). Heterologous E. granulosus antigens (EgHF and EgAgB) showed low sensitivities (43 and 50%, respectively) with high numbers of unspecific reactions (>16%) in the control group. In the follow-up of four dogs, the serological reaction patterns against the antigens Em2G11, EmP and EmAd/I showed some correlations with the development of the parasite mass. Intestinal infections cannot be differentiated from AE by most antigens (except EmII/3–10) and have to be excluded by the absence of coproantigens or DNA of E. multilocularis in faecal samples.

Taenia solium metacestode, a larval pork tapeworm, is a causative agent of neurocysticercosis, one of the most common parasitic diseases in the human central nervous system. In this study, we identified a cDNA encoding for a cathepsin L-like cysteine protease from the T. solium metacestode (TsCL-1) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1216 bp encoded 339 amino acids with an approximate molecular weight of 37.6 kDa which containing a typical signal peptide sequence (17 amino acids), a pro-domain (106 amino acids), and a mature domain (216 amino acids). Sequence alignments of TsCL-1 showed low sequence similarity of 27.3-44.6 to cathepsin L-like cysteine proteases from other helminth parasites, but the similarity was increased to 35.9-55.0 when compared to mature domains. The bacterially expressed recombinant protein (rTsCL-1) did not show enzyme activity; however, the rTsCL-1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. It degraded human immunoglobulin G (IgG) and bovine serum albumin (BSA), but not collagen. Western blot analysis of the rTsCL-1 showed antigenicity against the sera from patients with cysticercosis, sparganosis or fascioliasis, but weak or no antigenicity against the sera from patients with paragonimiasis or clonorchiasis.

Neutrophils, eosinophils and macrophages interact with invading parasites and naive hosts. The initial reaction of leukocytes is the generation of reactive oxygen species (ROS). The cytotoxic effects of extracts derived from intact Cysticercus cellulosae and from the scolex or membrane fractions on neutrophils were examined. DNA fragmentation of neutrophils was observed when cells were incubated with an extract from the intact metacestode; however, the addition of antioxidant enzymes to the incubation medium had a protective effect. The scolex and membrane extracts did not affect DNA fragmentation of neutrophils. Hydrogen peroxide production of neutrophils incubated with metacestode fractions from C. cellulosae increased by 190% (total extract), 120% (scolex) or 44% (membrane). An increase in antioxidant catalase activity (28%) concomitant with the increased production of ROS was observed in neutrophils incubated with metacestode fractions, which could be an attempt at self-protection. ROS production by neutrophils in the presence of the intact cysticerci extract did not alter phagocytosis. In contrast, the scolex and membrane fractions increased the phagocytic capacity of neutrophils by 44 and 28%, respectively. The results showed that the extract from intact C. cellulosae was toxic for neutrophils via ROS production, leading to DNA fragmentation and inhibition of phagocytic capacity, but neutrophils are able to protect themselves against oxidative stress by via catalase activity.

Immunity to Taenia solium infection was investigated using an experimental intramuscular oncosphere infection assay (IMOA) model in pigs. Three naturally infected pigs with cysticercosis were treated with oxfendazole (OFZ), a drug demonstrated to kill cysts in porcine muscle. These animals were then challenged with oncospheres but did not develop any cysts while three uninfected pigs that were similarly challenged, did develop intramuscular cysts. In another study, two groups of three pigs each were immunized with crude T. solium oncosphere and metacestode antigens, respectively, and tested with the IMOA. Immunization with crude oncosphere antigens (OAs) induced 100% protection, while metacestode antigens provided only partial protection. Immunoblots showed that pigs with complete immune protection to oncosphere intramuscular challenge had antibodies to two OAs at 31.3 and 22.5 kDa, respectively. Antibody to these two antigens was absent in pigs immunized with metacestodes or in uninfected control pigs. This study demonstrated the presence of two antigens that are unique to the oncosphere. Although, antibody to these two antigens is consistently present in pigs that are protected from an oncosphere intramuscular challenge their role in preventing infection by T. solium larval cysts is still hypothetical.

Human alveolar echinococcosis (AE), caused by the metacestode stage of Echinococcus multilocularis, is a serious zoonosis which caused up to 100% lethality in untreated patients before the 1970s, when modern methods of treatment were not yet established. AE occurs in large areas of the northern hemisphere mostly with low country-wide prevalences, but high prevalences of up to 4% have been reported from small population groups in highly endemic foci, e.g. from China. AE includes many veterinary aspects which are the topic of this review. Recent studies have shown that E. multilocularis has a wider geographic range than previously anticipated. There is evidence for growing populations of red foxes (Vulpes vulpes) in some areas, for increasing invasion of cities by foxes and also for establishment of the parasite cycle in urban areas. These and other factors may lead to an increased infection risk for humans. Significant progress has been made in the development of sensitive and specific new techniques for the intra vitam and post mortem diagnosis of intestinal E. multilocularis infection in definitive hosts, notably the detection of coproantigen by enzyme-linked immunosorbent assay and of copro-DNA by PCR. Both tests can also be used for the identification of E. multilocularis in faecal samples collected in the environment. Recommendations are given for chemotherapy and chemoprophylaxis of the intestinal infection in definitive hosts. In recent years, infections with the metacestode stage of E. multilocularis have not only been diagnosed in humans in several regions, including at least eight countries in central Europe, but also in animal species which do not play a role in the transmission cycle (wild and domestic pigs, dogs etc.). From 1987 to 2000 our group in Zurich has diagnosed 10 cases of AE in dogs and 15 in captive monkeys. In 2 dogs, concurrent infections of the intestine and of the liver with adult and larval stages of E. multilocularis, respectively, were observed for the first time. Clinical data are presented, and methods of diagnosis and treatment (surgery, chemotherapy) are described. Furthermore, small liver lesions caused by E. multilocularis were diagnosed in 10% of 90 slaughter pigs, and 2.9% of 522 breeding sows had specific serum antibodies against parasite antigens. In view of the unpredictable epidemiological situation, all possible measures for preventing E. multilocularis infections in humans and in domestic animals should be initiated by the veterinary and health authorities.

Taenia solium cysticercosis is still a serious public health problem in several countries where poverty and lack of hygiene favor transmission. Because pigs are the primary intermediate hosts, prevalence of porcine cysticercosis is a reliable indicator of active transmission zones. Serological diagnostic methods are important tools for epidemiological studies since they can be applied to living animals on a large scale. Four antigen preparations (cyst fluid and crude) from T. solium and T. crassiceps metacestodes were compared for swine cysticercosis diagnosis by indirect ELISA (IE). Twenty-eight serum samples from swine naturally and experimentally infected by cysticerci of T. solium and 56 serum samples from swine reared in commercial herds were tested. Best results of overall sensitivity were obtained by the use of cyst fluid and crude antigen of T. crassiceps metacestode (100 and 96.4%, respectively). Using homologous antigen preparations we have observed higher specificity percentage (98.2% for cyst fluid and 96.4% for crude metacestode T. solium antigen). We concluded that sensitivity is of far more importance than specificity for identification of endemic areas in order to prevent transmission to man. We conclude, therefore, that IE performed with cyst fluid antigen of T. crassiceps metacestode is a better tool for that purpose.

The Spanish sheep and horse strains of Echinococcus granulosus possess several differential characteristics in their metacestode stage. Cysts from sheep vary widely in size and fertility, but they usually have a thin cyst wall and, when fertile, a whitish hydatid sand formed by brood capsules and protoscoleces. Two types of infections have been observed in horses: one resembling that of sheep, caused by small, non-fertile cysts with a thin wall, and a second type caused by medium to large, always fertile cysts with a thick wall. In this latter case, hydatid sand is always dark brown in color and formed mainly by brood capsules (with almost no free protoscoleces) and abundant calcareous corpuscles. These characteristics of the fertile equine cysts, which were identified in other studies by genetic, biochemical, immunological and physiological criteria as belonging to the horse strain, have not been previously described nor observed in cysts from other host species. It is considered that the horse strain possesses a strong intermediate host specifity.

Adult female Swiss-Webster mice were immunized either intraperitoneally (IP) or subcutaneously (SQ) with cyst fluid or a genetically engineered fusion protein, Taenia crassiceps antigen 2-maltose binding protein (TCA2-MBP) from Taenia crassiceps metacestodes, or with live, non-budding cysts SQ, and then challenged IP with T. crassiceps metacestodes and necropsied 9 weeks later. Numbers of peripheral blood eosinophils were increased after IP immunization, but were not increased after SQ immunization or with SQ cysts given before the challenge infection. Eosinophil numbers gradually decreased over the course of the experiment, and were not found in increased numbers in the blood or peritoneal cavity at necropsy. Antigen-specific antibody responses were seen at day 14 or 28 in IP and SQ immunized groups; IgG1 and IgG3 isotypes continued to increase over the course of the experiment, A significant protective response was induced by immunization with the cyst fluid (15 ± 4, X ± SE recovered larvae) or the TCA2-MBP (22 ± 22) given IP, but not SQ (122 ± 36; 207 ± 53, respectively) as measured by the numbers of larvae recovered at necropsy. Live cysts given SQ resulted in reduced numbers of cysts in the peritoneal cavity (188 ± 66), but was not as effective as cyst fluid or TCA2-MBP given IP. Locally (IP) induced immune responses may be involved in the development of the protective response to a challenge infection with T. crassiceps metacestodes.

C. Larralde et al. (1990, Aoff. Pathol. Lab. Med., 114:926-928) demonstrated that heterologous antigen from the laboratory-adapted murine Taenia crassiceps metacestode may substitute those from Taenia solium in the immunodiagnosis of human cysticercosis by the indirect enzyme-linked immunosorbent assay (IE). This antigen is easily obtained at a laboratory level and solves the problem of T. solium cysticerci collection from naturally or experimentally infected swine. In this study an IE employing a heterologous antigen from the T. crassiceps metacestode was evaluated for the immunodiagnosis of swine cysticercosis. Sera from 300 swine free of T. solium cysticerci by post-mortem examination were employed to determine two IE cut-off values: 1) Mean ELISA values + 2 standard deviations (2 sigma cut-off) and 2) - Mean ELISA values + 3 standard deviations (3 sigma cut-off). The specificity of IE was 97% with the 2 sigma cut-off and 100% with the 3 sigma cut-off. When applied to ten sera from swine infected by cysticerci of T. solium by post-mortem examination, the sensitivity of IE was 100% independent of the cut off.

The present cytochemical study was undertaken to provide more information on the localization of enzymatic and glycoconjugates in the germinal membrane of the Echinococcus granulosus cyst. The distinctive distribution of binding sites for two lectins (peanut agglutinin and Dilochos biflorus agglutinin) in the germinal membrane are described. An investigation is made of the distribution and specific activity of adenosine triphhosphate, alkaline phosphatase and acid phosphatase. The results suggest that cells located in the deeper layer of the germinal membrane are intrinsic in the cellular differentiation process. The dissimilares detected in both the enzymatic activity and the lectin-binding receptors could be associated with metacestode development or degeneration.

Of 48 lambs slaughtered at a local abattoir, 60.4% were either found to be infected with Taenia hydatigena cycts, or showed evidence of abortive infections (white spots on the liver) with that parasite. The micro ELISA test and the IHA test did not distinguish between animals showing evidence of infection at slaughter and those which did not. Both tests gave results which correlated significantly with each other. The antigens used in the tests cross-reacted with each other, which may be the reason for the large number of false negative results.

Excretory—secretory and somatic preparations of Taenia taeniaeformis contained shared immunologically-active components although immunoelectrophoresis indicated that the major antigen present in the excretory—secretory preparations was only a minor component of the somatic preparations. Both antigens gave similar immunoelectrophoretic reactions with sera from mice infected with either T. taeniaeformis or Taenia crassiceps , but there was evidence from results using the ELISA technique tha the excretory—secretory components showed more species specificity than thsoe of somatic origin.

Two trials were carried out to investigate the protective value of various antigens in the immunization of lambs against Taenia multiceps metacestode infections. The first trial involved groups of 5 or 6 lambs immunized with activated T. multicep oncospheres; activated T. hydatigena oncospheres; T. multiceps coenurus fluid antigens, and two untreated control groups. All except one of the control groups were challenged orally with T. multiceps oncospheres 5 weeks later. The results of the trial were equivocal with 1 to 3 lambs from each group showing post-mortem evidence of infection. There was evidence that some lambs from this trial had previously been exposed to Taenia oncospheres, thus nullifying possible effects of immunization. A second trial used two groups of 5 “cestode free” lambs. One group was immunized with T. multiceps oncosphere-derived antigens prior to challenge and showed significant protection when compared with the shamvaccinated controls.

Taenia saginata cyst fluid was examined for host proteins; IgG1 and IgG2 as well as haemolytic complement activity were detected. Polyacrylamide gel electrophoresis revealed differences in proteinograms among the samples taken from 1-, 4-, and 10-month old cysts. Fluid from older cysts had fewer protein components and showed a weaker antigenic reaction with sera of bovines infected with T. saginata than that of younger cysts. The roles of antibody and complement in initiating degeneration of the parasite are discussed.