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The tapeworm Dipylidium caninum is the most widely distributed cestode infecting dogs, cats, and sometimes humans, worldwide. The diagnosis of the infection caused by D. caninum is achieved via the visualization of proglottids in feces or with traditional microscopic tests, but both lack sensitivity. The present study has evaluated and compared the diagnostic performance of a PCR protocol on different feline biological samples to detect D. caninum. A sample of feces, a Scotch tape test from the perianal area, and a rectal swab were collected from a total of 100 privately owned cats from Italy and Greece. All fecal samples were subjected to macroscopic examination and to floatation. Based on the results of the above tests the cats were divided in three groups, i.e. (i) cats positive for D. caninum (regardless of positivity for other endoparasites (Group A; n = 50 cats), (ii) cats negative for D. caninum but infected by other helminths (Group B; n = 25 cats), and (iii) cats negative for intestinal endoparasites (Group C; n = 25 cats). For each sample, the DNA was extracted from feces, floatation supernatant, Scotch tape test and rectal swabs and subjected to PCR. For 33 cats from Group A, at least one sample type scored positive at PCR. Of these, all were PCR-positive in the floatation aliquot, while nine and one cats were positive by PCR on feces and Scotch tape test, respectively. Swabs were negative by PCR for all the cats. None of the samples from cats of Groups B and C was positive by any PCR. Sequences obtained from amplicons generated from samples of cats enrolled in Italy had 99-100 % identity with the recently described D. caninum feline genotype. The data presented here suggest that PCR could be a useful tool for diagnosing D. caninum infections, under certain circumstances, e.g. when proglottids are unidentified, unseen or overlooked, even though it has limitations, e.g. false negative results due to fecal PCR inhibitors, uneven distribution of parasitic elements, or to intermittent proglottid and/or egg shedding. Thus, it may not be, currently, the best diagnostic choice for dipylidiosis., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Simone Morelli reports financial support was provided by IDEXX Laboratories Inc. Simone Morelli reports a relationship with IDEXX Laboratories Inc that includes: funding donations. This Study has been supported with a donation to the University of Teramo from IDEXX Laboratories, of which MB and JB are employees, and KD was employee at the time of the Study. No funding number is available. In the past years DT and AdC have acted as occasional consultants for IDEXX Laboratories. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Tapeworm infection in horses can cause serious health concerns, and recent data have documented treatment failures in the most common species, Anoplocephala perfoliata. The threat of anthelmintic resistance in A. perfoliata is of particular concern because of poor diagnostic performance of standard egg counting techniques for detecting this parasite. This study compared the performance of three diagnostic techniques 1) Mini-FLOTAC, 2) Cornell-Wisconsin, and 3) Proudman and Edwards used to detect and quantify A. perfoliata eggs in naturally infected horses. Eighteen adult female horses from the University of Kentucky's historic parasitology herd were included in this study. Fecal samples were collected from all horses at five collection time points two weeks apart and analyzed with the three techniques. A total of 90 samples were collected and 270 counts determined in the study. The proportions of positive samples determined by the three techniques were significantly different from each other (p<0.05): Mini-FLOTAC (16%), Cornell-Wisconsin (47%), and Proudman and Edwards (70%). The Proudman and Edwards technique counted consistently higher numbers of tapeworm eggs compared to the other two techniques throughout the study [p < 0.05]. Total raw counts of tapeworm eggs across the study for each technique were 16, 88, and 410 for the Mini-FLOTAC, Cornell-Wisconsin, and Proudman and Edwards, respectively. This study demonstrated that the Proudman and Edwards technique was superior in diagnosing A. perfoliata infection. Future work needs to assess this technique's potential for Fecal Egg Count Reduction Testing (FECRT)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Intestinal parasites, including cestodes like Dipylidium caninum, are common in dogs in the United States of America (USA), but fecal flotation consistently, and, at times, dramatically, fails to identify many of these infections. To determine the extent to which including coproantigen testing for D. caninum would improve the identification of dogs infected with this cestode, we evaluated fecal samples from 877 dogs (589 pet and 288 from municipal shelters) from six USA states using zinc sulfate (specific gravity 1.24) fecal flotation with centrifugation along with coproantigen detection for Giardia sp., hookworms, ascarids, and Trichuris vulpis. For D. caninum, PCR of perianal swabs was included. Intestinal parasite infections were identified, using centrifugal fecal flotation or coproantigen, in 265 dogs (13.2 % pet, 64.9 % shelter). Dipylidium caninum infection was detected in 5.6 % of dogs with the combination of coproantigen and centrifugal fecal flotation, and 7.3 % of dogs when perianal swab results were included; prevalence varied by diagnostic method, population, and geographic region. In pet dogs, D. caninum infection was identified by fecal flotation (0), coproantigen (2.2 %), or perianal swabs (1.2 %). The same methods revealed infection in 0.3 %, 12.5 %, and 11.1 % of shelter dogs, respectively. Frequent use of praziquantel in shelter dogs (116/288; 40.3 %) may have reduced prevalence. Positive and negative agreement of D. caninum coproantigen with perianal swab PCR in pet dogs was 85.7 % and 98.8 %, respectively. Multiple logistic regression analysis accounting for region, population, and age found D. caninum infection to be more common in shelter dogs relative to pet (adjusted OR 4.91 [2.48, 10.24]) and in the Southcentral and Southeast regions relative to North (adjusted OR 9.59 [1.92, 174.13] and 17.69 [3.67, 318.09] respectively). Coproantigen testing also enhanced the detection of other intestinal parasites over fecal flotation alone, including Giardia sp. (14.7 % vs 3.3 %), hookworms (13.8 % vs 8.4 %), ascarids (2.9 % vs 2.2 %), and T. vulpis (2.9 % vs 1.4 %). Together, these data indicate that the coproantigen assay employed increases detection of D. caninum infections several fold, supporting the use of this test in clinical practice, and add to a growing body of research documenting enhanced diagnosis through implementation of multiple laboratory-based methods., Competing Interests: Declaration of Competing Interest As employees of IDEXX Laboratories, Inc., JB, DE, RH, JH, PT, and MB have an affiliation to the commercial funders of this research. The other authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Chickens and ducks are important sources of essential proteins and nutrition for global consumption, especially their eggs and meat. Tapeworm infections in chickens and ducks are the cause of serious poultry health and economic problems in the processing of livestock and food production systems. Raillietina are cosmopolitan in distribution and are possibly the most common tapeworm parasites. There are three important species regarding avian infection, with different pathogenicity, including Raillietina echinobothrida, R. tetragona, and R. cesticillus. Co-infection diagnosis of these tapeworms using morphological analysis can be performed, but this is time-consuming and complicated. Therefore, this study aimed to develop a triplex PCR for the detection and discrimination of three Raillietina species. The triplex PCR assay specifically amplified target DNAs with no inter-specific interference and produced a specific band for each species. According to the specificity test, there was no cross-amplification with the DNA template of related parasites and their hosts. The lowest detectable DNA concentrations were evaluated and provided sensitivities of 0.5 pg/μL for R. echinobothrida, 5 pg/μL for R. tetragona, 50 fg/μL for R. cesticillus, and 5 pg/μL for the combination of DNA from all three species. Simultaneous detection limits of egg capsules and gravid proglottids was also performed, with and without feces. The interference of feces in the reaction was related to a decrease in sensitivity, but simultaneous detection of three Raillietina species in amounts lower than one gravid proglottid and ten egg capsules was still successful. Thus, this study is the first triplex PCR assay for Raillietina detection and can be utilized as an alternative diagnostic tool for the detection and discrimination of R. echinobothrida, R. tetragona, and R. cesticillus infection in poultry through the verification of fecal specimens. In addition, it could improve the performance of specific treatments and promote veterinary healthcare., (Copyright © 2022. Published by Elsevier B.V.)

We investigated the prevalence of equine tapeworms, Anoplocephala spp., in Danish horses during a seven-year period assessed by coprological analysis. The data material included >11,000 observations distributed over the period 2009-2015, and four major geographic regions in Denmark representing sandy soils and clayey moraine soils, respectively. This allowed us to test for possible differences between regions (or soil types) and seasons. For a sub-set of the data (about 1200 observations) age of horses was also known, and it was shown that prevalence was highest in the age group 1-5years and then declined with age. We observed no significant difference in the prevalence of Anoplocephala spp. across the geographic regions suggesting that risk of infection was largely the same irrespective the soil types included in the study. Autumn was the season with highest prevalence. However, seasonality was not very marked, and our results show that egg-producing Anoplocephala spp. are found in Danish horses year-round., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The current study evaluated helminth infections in birds raised in different production systems for different purposes (extensive/dual-purpose, semi-intensive/broiler, semi-intensive/hen, intensive/hen and intensive/broiler) in Brazil. A total of 374 birds was assessed for helminths at necropsy using standard parasitological methods. During the necropsies, organs from the gastrointestinal tract (crop, esophagus, proventriculus, gizzard, small intestine, large intestine and ceca) of each bird were collected and the contents fixed in 70% ethanol. Additionally, the trachea and eyes were assessed for the presence of helminths. The small intestine was examined using a methodology that allowed the recovery of cestode scolices attached to the intestinal mucosa. Stereomicroscopy and optical microscopy were used to detect and identify helminth species based on their morphological characteristics. Fifteen helminth species were found among birds from the different systems. The extensive system presented the highest number of helminth species (six cestodes, seven nematodes and one trematode) and the highest number of parasites (mean helminths/bird), followed by the semi-intensive system (broiler: six cestode and four nematode species; hens: five cestode and three nematode species). Hens from the intensive system were parasitized by five cestode, four nematode and one trematode species. No parasites were detected in broilers raised in the intensive systems. The results obtained in this study highlight the need for special attention and the implementation of biosecurity measures for the prevention of helminth infections in intensive systems (hens) and particularly in extensive and semi-intensive alternative poultry production systems., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Mesocestoides corti is a widely used model for the study of cestode biology, and its transition from the larval tetrathyridium (TT) stage to the strobilated, adult worm (ST) stage can be induced and followed in vitro. Here, a proteomic approach was used to describe and compare M. corti TT and ST protein repertories. Overall, 571 proteins were identified, 238 proteins in TT samples and 333 proteins in ST samples. Among the identified proteins, 207 proteins were shared by TTs and STs, while 157 were stage-specific, being 31 exclusive from TTs, and 126 from STs. Functional annotation revealed fundamental metabolic differences between the TT and the ST stages. TTs perform functions related mainly to basic metabolism, responsible for growth and vegetative development by asexual reproduction. STs, in contrast, perform a wider range of functions, including macromolecule biosynthetic processes, gene expression and control pathways, which may be associated to its proglottization/segmentation, sexual differentiation and more complex physiology. Furthermore, the generated results provided an extensive list of cestode proteins of interest for functional studies in M. corti. Many of these proteins are novel candidate diagnostic antigens, and/or potential targets for the development of new and more effective antihelminthic drugs., Biological Significance: Cestodiases are parasitic diseases with serious impact on human and animal health. Efforts to develop more effective strategies for diagnosis, treatment or control of cestodiases are impaired by the still limited knowledge on many aspects of cestode biology, including the complex developmental processes that occur in the life cycles of these parasites. Mesocestoides corti is a good experimental model to study the transition from the larval to the adult stage, called strobilation, which occur in typical cestode life-cycles. The performed proteomics approach provided large-scale identification and quantification of M. corti proteins. Many stage-specific or differentially expressed proteins were detected in the larval tetrathyridium (TT) stage and in the strobilated, adult worm (ST) stage. Functional comparative analyses of the described protein repertoires shed light on function and processes associated to specific features of both stages, such as less differentiation and asexual reproduction in TTs, and proglottization/segmentation and sexual differentiation in ST. Moreover, many of the identified stage-specific proteins are useful as cestode developmental markers, and are potential targets for development of novel diagnostic methods and therapeutic drugs for cestodiases., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Genetic and morphological diversity of Thysaniezia tapeworms from cattle and sheep in Senegal was investigated using light and scanning microscopic observations and molecular analysis based on mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear small subunit ribosomal RNA gene (SSU rDNA). A total of 52 adult tapeworms were collected from sheep and cattle. Although the tapeworms of the two hosts were morphologically very close, phylogenetic analysis based on cox1 and SSU rDNA gene sequences showed that they were divided into two clades corresponding each to a host. The maximum pairwise divergence between the clades were 12.1% in cox1 and 2.9% in SSU rDNA, indicating they are distinct species. The tapeworms collected from sheep were morphologically identified as Thysaniezia ovilla, a cosmopolitan species in domestic ruminants. Detailed morphological observations revealed a consistent difference between the tapeworms obtained from sheep and those from cattle. The latter were identified as Thysaniezia connochaeti. The present study highlights presence of two species of Thysaniezia among domestic ruminants in Senegal: T. ovilla specific to sheep and T. connochaeti specific to cattle. Our work is the first report of T. connochaeti from domestic animals., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Although several techniques exist for the detection of equine tapeworms in serum and feces, the differential diagnosis of tapeworm infection is usually based on postmortem findings and the morphological identification of eggs in feces. In this study, a multiplex polymerase chain reaction (PCR)-based method for the simultaneuos detection of Anoplocephala magna, Anoplocephala perfoliata and Anoplocephaloides mamillana has been developed and validated. The method simultaneously amplifies hypervariable SSUrRNA gene regions in the three tapeworm species in a single reaction using three pairs of primers, which exclusively amplify the internal transcribed spacer 2 (ITS-2) in each target gene. The method was tested on three types of sample: (a) 1/10, 1/100, 1/500, 1/1000, 1/2000 and 1/5000 dilutions of 70 ng of genomic DNA of the three tapeworm species, (b) DNA extracted from negative aliquots of sediments of negative control fecal samples spiked with 500, 200, 100, 50 and 10 eggs (only for A. magna and A. perfoliata; no A. mamillana eggs available) and (c) DNA extracted from 80, 50, 40, 30, 10 and 1 egg per 2 μl of PCR reaction mix (only for A. magna and A. perfoliata; no A. mamillana eggs available). No amplification was observed against the DNA of Gasterophilus intestinalis, Parascaris equorum and Strongylus vulgaris. The multiplex PCR method emerged as specific for the three tapeworms and was able to identify as few as 50 eggs per fecal sample and as little as 0.7 ng of control genomic DNA obtained from the three species. The method proposed is able to differentiate infections caused by the two most frequent species A. magna or A. perfoliata when the eggs are present in feces and is also able to detect mixed infections by the three cestode species., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Current copro-diagnostic tests for Anoplocephala perfoliata show high variation in their sensitivity and given the morphological similarity of Anoplocephala spp. eggs, this could be related to the presence of Anoplocephala magna alone or co-existing with A. perfoliata. In the present study, coprology was significantly more sensitive (p<0.01) at detecting A. magna than A. perfoliata. This difference was independent of the parasite burden and was greater when testing was limited to horses with mature or gravid tapeworms. A. magna infection was strongly linked to young horses (≤ 2 years). The eggs of A. magna are smaller. Using 15 and 70 μm cut-offs for oncosphere diameter and the major shell bisector length, respectively, the eggs of A. perfoliata were identified with 100% sensitivity, 97% specificity and 98% sensitivity, 84% specificity. The use of these two morphometric variables would therefore be useful for the copro-identification of A. perfoliata in countries where both species coexist., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Canine peritoneal larval cestodiasis (CPLC) is an unusual parasitic disease in dogs that is caused by asexual proliferation of larval Mesocestoides. A 12 year-old spayed Shetland sheepdog with abdominal distension was referred to the Animal Medical Center at Nihon University, Japan. The presence of ascites was confirmed by abdominal ultrasonography and X-ray imaging. In addition, a number of parasites were observed in the ascitic fluid collected by abdominal paracentesis. Each of the whitish colored parasites was less than 1mm in size. The parasites were morphologically identified as Mesocestoides sp. tetrathyridia. The parasites had four suckers and calcareous corpuscles, but no hooks or rostellum. Mitochondrial (mt) 12S rDNA and mt cytochrome c oxidase subunit 1 DNA amplified from the tetrathyridia were used for molecular identification to species level. DNA sequence analysis showed that the tetrathyridia shared more than 99% identity with M. vogae (syn. M. corti) for each gene. The patient was treated with a standard dose (5mg/kg) of praziquantel, which was administered subcutaneously twice at an interval of 14 days. This resulted in successful deworming. This is the first case that CPLC was diagnosed in a dog that had never been taken outside of Japan, indicating that M. vogae is distributed in this country., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Dipylidiasis is a zoonotic parasitic infestation caused by the dog tapeworm Dipylidium caninum. Human dipylidiasis has been rarely reported in English literature. Young children are mostly at risk of acquiring the infection due to their close association with dogs and cats. We report a rare case of Dipylidium caninum infection in a 4 year old male child. The diagnosis was based on microscopic examination of stool. Confirmation of the proglottid segments was done by histopathological examination. To the best of our knowledge this is the first human case of Dipylidium caninum reported from this part of the country.

Many cestodes are capable of invading the central nervous system (CNS), and several are highly prevalent in the developing world. Neurocysticercosis due to Taenia solium and echinococcosis due to Echinoccocus granulosus are two of the most common parasitic infections affecting humans, but other less well-known parasites can also infect the nervous system. Coenurosis, caused by Taenia spp. such as T. multiceps, T. serialis, or T. brauni; sparganosis, caused by Spirometra spp., and neurocysticercosis caused by T. crassiceps are three less frequent zoonotic conditions that should be considered in the differential diagnosis of patients presenting with CNS infection - especially if they have lived in or traveled through areas where these infections are endemic. Diagnosis of these infections is typically made through a combination of serological testing, histopathology, and neuroimaging., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The tapeworm responsible for equine colic, Anoplocephala perfoliata, is considered the most common intestinal tapeworm of horses worldwide. However, there is evidence that Anoplocephala magna has a similar prevalence in North America and Spain, and possibly in other countries, highlighting the need for diagnostic methods capable of distinguishing between these two species. Currently, immunodiagnosis of A. perfoliata is based on the identification of the 12/13 kDa excretory/secretory (E/S) A. perfoliata immunoreactive antigen, which while apparently specific, has never been tested in sera from A. magna-positive horses. Accordingly, we evaluated the specificity of 12/13 kDa E/S A. perfoliata antigen for the first time by testing this crude antigen against A. magna-positive sera in Western blot. In addition, we characterized a somatic (Som) crude antigen of A. perfoliata and for the first time, the E/S and Som crude antigens of A. magna, evaluating their potential utility for the differential serodiagnosis of equine anoplocephalosis in sera from horses of known parasitic status. SDS-PAGE revealed major low MW bands at: 14 and 12 kDa for E/S and Som-A. magna; 14 and 11 kDa for E/S A. perfoliata; and 11 and 10 kDa for Som-A. perfoliata. Protein regions at 12-14 kDa (E/S A. perfoliata), 10-15 kDa (Som-A. perfoliata) and 10-12 kDa (Som-A. magna) were recognized by Anoplocephala-positive sera at the genus but not the species level. These findings demonstrate cross-reactivity of these unpurified antigenic components, precluding their use in differential diagnosis between A. perfoliata and A. magna. Although these results do not directly indicate cross reactivity at the purified 12/13 kDa component of the E/S A. perfoliata antigen, it is possible that current immunodiagnostic methods based on this component might not accurately differentiate between these two tapeworm species, suggesting erroneous diagnosis of A. perfoliata in areas where A. magna is present., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Coenurosis is a disease of the central nervous system in sheep, caused by Coenurus cerebralis, the larval stage of Taenia multiceps, a tapeworm, which infests the small intestine of carnivores. In 80-90% of cases, the cyst is located in one cerebral hemisphere, whilst in 5-10% of cases, it is localised in the cerebellum; rarely it involves two sites in the brain of the affected animal. Listeriosis, louping-ill, sarcocystosis and polioencephalomalacia and brain abscessation should be considered when formulating a diagnosis of acute coenurosis. In all cases, it is essential to carefully examine the animal and not simply rely on results of ancillary tests (mainly of cerebrospinal fluid examination), as disorders other than coenurosis can be responsible for changes in the results of these tests. Treatment is based on surgical removal of the coenurus cyst after general anaesthesia of the animal; the approach has a very good success rate, especially after accurate localisation of the lesion. Despite that, many farmers may choose to slaughter those sheep fit for marketing for economic reasons and euthanise those in poor condition., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Accurate diagnosis of parasites within individual hosts remains a difficult task. Incorrect and/or inaccurate diagnosis restricts the potential for targeted treatment of individuals and limits our understanding of key epidemiological characteristics of potential pathogens of domestic stock. In this study, we compared the specificity and sensitivity of four diagnostic methods for determination of the presence and intensity of the cestode Anoplocephala perfoliata in horses. Over 400 horses from an abattoir in south-central Alberta were evaluated for the presence and number of cestodes. Thirty one horses were infected with 1 to >1000 worms. Diagnosis based upon faecal egg counts of horses with known numbers of worms was least accurate in detecting worm presence. Detection of circulating antibodies to the cestode was most sensitive using Western blot analysis (100%), but had lower specificity (87%). A serum-based ELISA had a lower sensitivity (70%) for detection of antibodies. A coproantigen ELISA had 74% sensitivity and 92% specificity, and there was a positive correlation between antigen concentration and tapeworm intensity. The coproantigen ELISA may have the potential as a diagnostic tool for determining the presence and intensity of this potentially pathogenic cestode.

This report describes the finding of Spirometra erinacei Faust, Campbell & Kellog, 1929 (Cestoda, Diphyllobothridae) infecting the small intestine of two Andean hog-nosed skunks (Conepatus chinga Molina, 1782), collected from the locality "Abra La Raya", at Cusco, Peru. Four cestodes were studied and identified as S. erinacei. This is the first report showing that the Andean hog-nosed skunk is one of the natural hosts for this parasite.

Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a combination of ivermectin and praziquantel. Of 42 horses subjected to coprological examination, 16 and 26 resulted negative and positive, respectively for the presence of A. perfoliata eggs at the coprological examination. The 26 coprologically positive animals were also positive by nested PCR. Fifteen out of the 16 horses coprologically negative were negative at the molecular assay, while one yielded a PCR product detectable on an agarose gel. Eighteen out of 26 positive horses were treated with a combination of ivermectin 18.7 mg/g and praziquantel 140.3mg/g and resulted subsequently negative by coprology and nested PCR performed 2 weeks after treatment. All infected and untreated animals had a high ELISA test optical density indicating high infection intensity and associated risk of colic. However, high optical density values were also obtained in four horses post-treatment and in three horses that were negative on molecular and coprological analysis. The results of the present work indicate that the nested PCR assay represents a valid method for the specific molecular detection of A. perfoliata in faecal samples collected from naturally infected horses and may have advantages over coprological and serological approaches for diagnosing A. perfoliata infection.

A 13-year-old Dalmatian dog was presented with a history of abdominal enlargement and reduced appetite for several months. Acute clinical signs were anorexia, vomiting and diarrhoea. During exploratory laparotomy, acute intestinal perforation due to a foreign body and peritonitis was diagnosed. In addition, the abdominal cavity was filled with multiple small (0.5 cm), white, cyst-like structures. Histopathology revealed typical cestode structures of the cyst walls but no protoscolices were found. PCR was performed with cestode specific primers of the mitochondrial 12S rDNA. The sequence showed a 99.75% identity with a Mesocestoides lineatus isolate published in the NCBI GenBank. This is the first case of canine peritoneal larval cestodosis (CPLC) in Germany and the first evidence of M. lineatus as causal agent for CPLC.

Anoplocephala perfoliata has a prevalence as high as 60% in some geographical areas and has been associated with increased risk for serious conditions in the horse including bowel irritation, ileal impaction, and spasmodic colic. Identification of infected animals based upon detection of eggs in feces is labor intensive and unreliable. This study involved the development of a test for A.perfoliata coproantigen using an antigen capture enzyme linked immunosorbent assay (ELISA) and correctly distinguished between infected and uninfected animals in a trial with a small sample size.

The diagnosis of tapeworm infections in horses relies on copro-diagnostic methods, which are time-consuming and of limited sensitivity for determination of the exact prevalence. The development of serological tests has slightly improved the detection of tapeworm infections, but more sensitive methods are still required. A polymerase chain reaction (PCR)-based approach may constitute a valuable tool to improve tapeworm diagnosis. Nuclear ribosomal DNA (rDNA) is a useful target for species and/or strain markers. Partial 18S, the internal transcribed spacer 1 (ITS-1), the 5.8S, the internal transcribed spacer 2 (ITS-2), and partial 28S rDNA of the equine tapeworms Anoplocephala perfoliata and Anoplocephaloides mamillana were amplified and sequenced. The lengths and GC contents of the regions sequenced were 2087-2091bp and 49.35-49.69% for A. perfoliata, and 2110-2119bp and 49.15-49.32% for A. mamillana, respectively. Sequence alignment and comparison of both taxa showed 79.3-80.2% identity. The lowest identities were found in the ITS regions with 39.9-43.5% for the ITS-1 and 59.5-61.2% for the ITS-2. No matches of the ITS-2 of A. perfoliata and A. mamillana were found with other species by BLAST search. For this reason, ITS-2 sequences seemed appropriate as accurate species markers and A. perfoliata ITS-2 primers were developed. The ITS-2 PCR enabled the detection of genomic DNA as low as 0.5 pgs. First efforts on the practical application of the PCR-based approach were made. A 6-mg fragment of a tapeworm proglottid was detected in 0.5 and 1g of faeces.

Two kinds of homogeneous proglottid, mature and gravid, of Dipylidium caninum were used as the antigens for immunodiagnosis of canine dipylidiosis in stray dogs in Tainan, Taiwan. The ELISA was performed on 30 serum samples; 24 from dipylidiosis, four from ancylostomosis and two from toxocariosis. The ELISA have specificity and sensitive of 100 and 50% for mature proglottid extract, and 75 and 100%, respectively, for gravid proglottid extract. EITB technique showed two major peptide bands of 94.8 and 97.9kDa were recognized in the sera pool of infected dogs.

The egg output and humoral antibody response to scolex antigens of the equine tapeworm Anoplocephala perfoliata were monitored in naturally infected foals by an egg flotation/centrifugation method and an indirect enzyme-linked immunosorbent assay (ELISA). The study was performed on a stud farm in south-western Sweden between May 1994 and April 1995. Sequential blood and faecal samples were taken from 21 foals during their first summer on pasture and until tapeworm eggs were detected. Results were expressed separately for 10 and 11 foals born before and after the end of April 1994, respectively. Increased levels of antibodies were noticed from October and onwards in both groups whereas tapeworm eggs were detected in the faeces of all foals about 4 months later. The antibody response was similar in both groups but it was more pronounced in foals born before April 1994. All foals were treated in March 1995 with an oral paste formulation of pyrantel pamoate at a dose rate of 38 mg/kg bodyweight. Most animals responded to the anthelmintic treatment and one month later, tapeworm eggs were only detected in one out of the 18 foals examined one month after treatment. Thus, the treatment reduced the number of A. perfoliata egg positive horses by 94%. A concomitant decline in antibody levels was also observed. Western immunoblot analysis of sequential individual serum samples showed that at least 10 different scolex antigens in the molecular weight range 10-200 kDa were recognised. Banding intensities, especially of the 10 kDa, 45 kDa and 66 kDa proteins were different in the sequential sera taken during the course of infection.

Whole worm extract (WWE) and excretory/secretory (E/S) antigens of Anoplocephala perfoliata were characterised by SDS-PAGE and their use in the serodiagnosis of equine cestodosis was evaluated. An enzyme-linked immunosorbent assay (ELISA) was used to compare WWE and E/S antigen as the capture layer in an antibody capture ELISA. E/S antigen gave the best differentiation between sera from tapeworm-positive and tapeworm-negative horses. The E/S-ELISA was optimised and validated against sera from horses of known tapeworm status. This assay gave a diagnostic sensitivity of 68% (n = 38) and a specificity in helminth-naive horses of 95% (n = 20). Western blot analysis of tapeworm-positive and tapeworm-negative sera revealed the immunoreactive proteins of E/S antigen to be those of 12 and 13 kDa.

A scolex antigen of the horse tapeworm Anoplocephala perfoliata containing at least 14 different proteins was employed in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to A. perfoliata in equine sera. The assay was applied to sera from 426 slaughtered horses with different numbers of worms and with varying degrees of intestinal lesions. As measured by the ELISA, there was a very strong effect on the antibody levels both from the number of tapeworms present and from the intestinal lesion score. However, considerable individual variation was observed between horses with similar worm counts. The ELISA values of horses that were either negative for strongyles and tapeworms or positive only for strongyles were similar, whereas both of these groups differed significantly from animals that harboured only A. perfoliata. Thus it seemed that cross-reactivity with concurrent nematode infections did not occur. Furthermore, a significant seasonal pattern in antibody levels was observed which reflected the establishment of newly acquired infections. No observations were made of confounding effects owing to age and previous anthelmintic treatment of the horses. It is concluded that the ELISA presented here has a potential for monitoring A. perfoliata infection on a herd level and can be used as a complementary diagnostic tool in epidemiological studies. The present study indicates that transmission of A. perfoliata in Central Sweden occurred during a short period in spring and more consistently in late autumn.

The authors reviewed the data acquired in ELISA technique for serological diagnosis of parasitosis. They reviewed the literature on Schistosomisis, Fasciolasis, Cestode infections (hydatidoses and cysticercoses) and nematode infections. They described their adaptation of the technique for detection of antimicrofilaria Loa loa antibodies and concluded that ELISA technique is reliable in the diagnosis of parasitic diseases even in areas where they are endemic.

A 72-year-old Filipino man who had emigrated to Canada had an abdominal infection with a proliferating acephalic larval cestode. Small, irregularly shaped parasites were within the substance of a lymph node, where they provoked a fibrogranulomatous and eosinophilic reaction. The parasite is described and is identified as an undifferentiated sparganum or tetrahyridium based upon comparison with two similar parasites from patients in Taiwan and Paraguay.