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Long-term dynamics of Ligula intestinalis and roach Rutilus rutilus: a study of three epizootic cycles over thirty-one years.

Data are presented on 2 full epizootic cycles and the start of a third of Ligula intestinalis in roach Rutilus rutilus in a small lake, and the relationships of these cycles to the densities of rudd, Scardinius erythrophthalmus, and Great Crested Grebes, Podiceps cristatus, over 31 years. The parasite was introduced to the lake by P. cristatus in 1973 at a time when the roach population had increased in response to eutrophication to a level at which individual fish growth was stunted and the hithero dominant rudd population had declined in numbers as a consequence of inter-specific competition with roach. Ligula prevalence peaked at 28% in only 2 years: thereafter parasite-induced host mortality caused a decline in the roach population, releasing fish from stunting and allowing the rudd population to recover. The consequent improved growth of roach individuals and their short life-span reduced Ligula transmission rates and prevalence levels declined to approximately 1% although Ligula nevertheless persisted for a further 10 years. Following a massive winter-kill of the fish populations in 1984-1985, fish and Ligula numbers declined to barely detectable levels and the parasite disappeared from samples. Rudd recovered first, then roach and interspecific competition again led to a decline in rudd numbers. This increase in roach numbers led to a decrease in roach growth rates, which coincided with the re-colonization of the lake by Ligula. This second epizootic of Ligula peaked within 2 years in 1991-1992, when up to 78% of roach were infected with a maximum abundance of 2.2 parasites and intensity of 21 parasites. Heavy parasite-induced mortality of roach led to a decline in numbers, an improvement in individual growth rate and a reduction of Ligula transmission rates such that the epizootic died out in 1996. Similar conditions of roach numbers and growth prevailed at the start of a third cycle in 1998. The course of events over the second cycle was so similar to that of the first that it confirms the interpretations of that cycle. Comparison with other localities shows that epizootics of Ligula always coincide with rapid increases in roach numbers, for whatever cause, and stunted growth, which together attract piscivorous birds. At the start of a cycle Ligula is a major determinant of the population dynamics of the roach, but at the end of the cycle the fish population dynamics determine those of the parasite. The cycles are not regulated and the roach-Ligula system is inherently unstable.

Animals↗

Detection of eosinophil differentiation factor and its relationship to eosinophilia in Mesocestoides corti-infected mice.

Eosinophil differentiation activity has been identified using a simple assay to detect eosinophil differentiation in vitro. Two factors were involved in this activity: eosinophil differentiation factor (EDF, Mr 32-64K) and IL-3 (Mr 19-44K). In this paper it is shown that eosinophil differentiation activity (EDA) can be detected in the serum of mice undergoing eosinophilia induced by Mesocestoides corti. This serum activity is shown to follow the ability of spleen cells to produce EDF after stimulation with parasite antigen or pokeweed mitogen. The activity in mitogen stimulated spleen supernatant (MSSS) and serum has a mean Mr of 38K (range 26K-58K). IL-3 is detectable in MSSS but not in serum. The appearance of the EDA is accompanied by an increase in eosinophil precursors in the bone marrow. These reach a peak at about eight to 16 days. A significant blood eosinophilia was detected by 16 days, reaching a peak at 24 days, although blood levels are a poor indicator of the number of eosinophils reaching the tissues. Eosinophils were present in large numbers in the spleen by 14 days and in the peritoneal exudate by 21 days. At peak levels, 5 X 10(7) eosinophils could be recovered from the peritoneal exudate.

Animals↗

Critical evaluation of taeniacidal antibiotic S15-1 (SQ 21, 704) for removal of natural tapeworm infections in dogs and cats.

The new taeniacidal antibiotic S15-1 (SQ 21,704) was evaluated against naturally occuring infections of Taenia pisiformis in 53 dogs, Dipylidium caninum in 35 dogs, T taeniaformis in 18 cats, and D caninum in 33 cats. It all instances, the compound was administered in gelatine capsules in a single oral dose. The doses tested were between and 200 mg/kg of body weight in dogs and between 15 and 45 mg/kg in cats. In dogs, doses of 25 mg/kg and greater were 100% effective against T pisiformis, whereas a dose of 50 mg/kg was necessary to clear D caninum. In cats, a single oral dose of 22.5 mg/kg was 100% efficacious against T taeniaeformis, and a single dose of 45 mg/kg (the largest dose tested) clearly seven of eight cats of D caninum. The efficacy was limited to tapeworms only; there was no efficacy against nematodes. The antibiotic was well tolerated by both species with no drug-related vomiting or other side-effects observed.

Administration, Oral↗

Oxfendazole: anthelmintic activity in calves artificially infected with six species of nematodes and in calves naturally infested with tapeworms.

In a series of 5 trials, in which 115 artificially infected calves were used, larval anthelmintic tests were done to assess the activity of oxfendazole at 4, 5 mg/kg against all 3 parasitic stages of 6 species of namatodes. An efficacy of over 98% was demonstrated against the 4th stage larvae (L4), 5th and adult stages of Haemonchus placei, Ostertagia ostertagi, Cooperia pectinata, Bunostomum Phlebotomum and Oesophagostomum radiatum and the 3rd larval stage (L3) of H. placei and C. pectinata. Against the L3 of O. ostertagi, B. phlebotomum and O. radiatum efficacies of 81,7%, 92,4% and 97,3% respectively were shown. Against Dictyocaulus viviparus adult, 5th, L4 and L3 efficacies were recorded of 100%, 96,7%, 95,4% and 89,1% respectively. A test for ovicidal activity confirmed that cattle can be safely moved to clean pasture 8 hours after dosing. Complete removal of strobilia of Moniezia benedeni from 25 naturally infested calves followed a dose rate of oxfendazole at 4,5 mg/kg, but of these each of 4 calves retained one scolex.

Animals↗

Catatropis hatcheri n. sp. (Digenea: Notocotylidae) from Heleobia hatcheri (Prosobranchia: Hydrobiidae) and notes on its life-cycle in Patagonia, Argentina.

A new species of Catatropis Oghner, 1905 from a freshwater Neotropical prosobranch snail, Heleobia hatcheri (Hydrobiidae), is described. Naturally infected snails were collected from Nahuel Huapí Lake in Andean Patagonia. The characteristics of the larval stages are also presented. Experimental adults were recovered from the distal region of the intestinal caeca of chicks and ducklings and natural adults from a wild duck Anas platyrhynchos. Adults of Catatropis hatcheri n. sp. can be distinguished from all other species of the genus in having 10-12 (11) ventral glands in each lateral row, the cirrus-sac extending back to between the first third and the middle of the body, the metraterm shorter than the cirrus-sac, a previtelline field of 1,258-1,544 (1,396), vitelline follicles reach back to the anterior border of the testes with some follicles extending slightly lateral to them, only external testicular margin lobed and genital pore in median line just posterior to the intestinal bifurcation. In addition, the eggs have one filament on each pole, the rediae contain one or two mature cercariae, and the cercariae are tri-oculate, with a long tail and encyst in the environment.

Animals↗

Contribution to the knowledge of helminthofauna of wild mammals of Somalia.

Within a survey of parasitic infections in wild mammals of Somalia, during the first semester of 1983 and the second of 1984, endoparasites were collected from 11 host species: Gazella soemmeringi, Gazella spekei, Madoqua saltiana, Phacochoerus aethiopicus, Xerus rutilus, Lepus sp., Genetta genetta, Herpestes (Galerella) sanguineus, Felis sylvestris libyca, Felis caracal, Canis mesomelas. A total of 22 species of helminths (20 nematode and 2 metacestode species) were identified. Lepus sp. is a new host for Heteroxinema (P.) proboscidiphora and Herpestes sanguineus for Oxinema sp. Teladorsagia hamata had not been previously reported in G. spekei; the genital cone and the corresponding supporting apparatus of this nematode are described. Two metacestodes were collected from the mesenteric membranes of a P. aethiopicus. The shape, number and length of their rostellar hooks, the size of the larvocysts, the occurrence of pseudostrobilum, are similar to those of the Taenia regis metacestode: but the size and shape of the hooks of the Somalian warthog larvocysts are slightly different and they have not been recovered from the musculature as previously reported in the intermediate hosts of T. regis.

Animals↗

Triaenophorus nodulosus and T. crassus in fish from northern Finland.

10,288 fish specimens of 34 species from three water bodies in northern Finland were studied mainly in 1978-1982 Triaenophorus nodulosus and T. crassus parasites. Six of the 31 species from the Bothnian Bay were found to harbour T. nodulosus plerocercoids, four of the 11 from the cold, oligotrophic Lake Yli-Kitka in northeastern Finland and only one of the 6 from the small, eutrophic Lake Kuivasjärvi. The ruff (Gymnocephalus cernuus) was most essential intermediate host in the lakes, as supported by stomach analysis of predator fish. Although the highest prevalences were found in adult burbot (Lota lota) in two of the areas, these fish are more likely a blind end in the life cycle of T. nodulosus in the present case. T. crassus larvae were found only in three coregonid species and once in the intestine of a lamprey (Lampetra fluviatilis) from the sea. Neither the prevalence nor the intensity of infection in most of the intermediate fish hosts of T. nodulosus was seen to increase with the length of the fish. The only exception was the ruff in Lake Yli-Kitka, where a sharp increase was encountered. No prominent seasonal variation in Triaenophorus larvae infections was found in any of the cases. The prevalence of T. nodulosus infection in its final host, the pike (Esox lucius), was highest in the sea (93%) and lowest in the eutrophic lake (47%). Seasonally, the lowest T. nodulosus infections were found in June-July in the two lakes. A clear maturation cycle was also found, with the youngest worms in the summer months in all cases. T. crassus was found in every third pike in the sea and its proportion in relation to T. nodulosus is higher in the smallest and largest fish. In only 12%t of the pikes did the number of T. crassus exceed that of T. nodulosus, however, the mean ratio being 1:13 to favour of T. nodulosus. The results give an indication that the composition of the fish fauna in each water body has a considerable influence on the T. nodulosus burden and its distribution among its potential intermediate fish hosts. With the most stable environmental conditions and the greatest variety of fish species, the parasite burden in the Bothnian Bay is most evenly distributed among the 6 intermediate hosts. The fewer fish species occur in the water body, as in the present eutrophic lake, the more the significance of a few or even one intermediate host will increase.

Animals↗

Murine hypodense eosinophils induce tumour cell apoptosis by a granzyme B-dependent mechanism.

PURPOSE: Eosinophils have been shown to potentiate anti-tumour cytotoxicity in both clinical and animal studies. The mechanism by which eosinophils induce tumour cell damage, however, has largely been speculative. The purpose of this study was to identify the mechanisms involved in eosinophil-induced tumour cell cytotoxicity. METHODS: To investigate eosinophil cytotoxicity, eosinophils were isolated from the peritoneal cavity of Mesocestoides corti-infected BALB/c mice, and were separated into normodense (ND) and hypodense (HD) populations using discontinuous Percoll density gradient centrifugation. The tumoricidal activity of ND and HD eosinophils was assessed using the [51Cr]-release cytotoxicity assay (a measure of cytolytic activity) and the JAM assay (a measure of apoptotic activity). Investigation of apoptosis-inducing molecules in HD eosinophils was undertaken by RT-PCR. The calcium chelator EGTA, serine protease inhibitor aprotinin and a competitive substrate for granzyme B were used to assess the role of perforin and granzyme B in HD eosinophil killing. RESULTS: Cytotoxic activity induced by HD eosinophils was significantly greater than that of ND eosinophils, and apoptosis was the principal killing mechanism. RT-PCR analysis revealed that HD eosinophils express mRNA for perforin, granzyme B and Fas ligand. Furthermore, HD eosinophil killing was markedly inhibited by EGTA, intracellular aprotinin and the granzyme B competitive substrate. CONCLUSIONS: These data are consistent with a hypothesis that murine HD eosinophils elicit tumoricidal activity via a granzyme B-dependent mechanism.

Animals↗

Effects of free and liposomized praziquantel on worm burden and antibody response in mice infected with Mesocestoides corti tetrathyridia.

The parasite burden in the liver and peritoneal cavity, and antibody levels directed to whole worm extract, have been monitored in serum from ICR-strain mice, infected orally with 55 tetrathyridia of Mesocestoides corti (Cestoda, Cyclophyllidea). The subcurative does (3x or 6x) of praziquantel (PZQ) (10 mg.kg-1 body weight) were administered to mice from day 14 post infection (p.i.) in two drug formulations: as PZQ suspended in Dorfman vehicle, or as PZQ incorporated in liposomes (lip.PZQ). The appearance of antibodies was time-dependent and correlated with the rate of reduction in numbers of tetrathyridia. PZQ in three and six doses caused the highest fall of parasite numbers in the liver on day 1 post therapy (p.t.). In the peritoneal cavity, a similar reduction in worm burden occurred but only after six doses of the drug. The worm count in the peritoneal cavity from groups of mice injected with lip.PZQ decreased most markedly on day 7 p.t., in the group treated with six doses of the drug. In the liver, the highest larvicidal effect, compared with the controls, was observed 6 days later (i.e. day 13 p.t.), following three doses of lip.PZQ. In all treated groups, two peak values of antitetrathyridial antibody levels were detected between days 1 and 13 p.t. (i.e. days 17 to 29 p.i.), after which there was a gradual but continuous increase in antibody tire.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

Treatment of fish parasites. 10. Effects of a new triazine derivative, HOE 092 V, on Monogenea: a light and transmission electron microscopy study.

For chemotherapy of fish parasitized by monogeneans, a novel triazine derivative, 2-[3,5-alpha-dichloro-4-(4-methyl-sulfonylphenoxy)-phenyl]-1-methy l- hexahydro-1,2,4-triazine-3,5-dion (HOE 092 V), was tested in vivo against the gill- and skin-parasitizing species Dactylogyrus extensus, D. vastator, and Gyrodactylus arcuatus. Naturally infected fish were incubated in aerated, separate tanks at 22 degrees C for 1, 2, 3, and 4 h in water containing 0, 1, 5, 10, or 15 micrograms HOE 092 V/ml, whereas Pseudodactylogyrus bini was tested in vitro at 10 micrograms HOE 092 V/ml for 2.25 h. As seen by means of transmission electron microscopy, in vivo treatment against D. extensus caused vacuolization and lysis of the parasite's tegument at a dose as low as 1 micrograms/ml over a 3-h exposure period. Higher doses, such as 5 and 10 micrograms/ml over the same exposure period, produced lesions in the circular and longitudinal musculature of D. extensus and differing degrees of damage to the ciliary cells of protonephridia and immature vitelline cells. There was 100% mortality in D. vastator when incubation was done with 10 micrograms HOE 092 V/ml for 4 h (G. arcuatus; 5 micrograms/ml for 4 h; 10 micrograms/ml for 1 h) and in P. bini after 2.25 h in vitro exposure. In all species tested, the anterior portion and the opisthaptor region were most sensitive to the drug action.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cecal rupture by Anoplocephala perfoliata infection in a thoroughbred horse in Seoul Race Park, South Korea.

A 7-year-old Thoroughbred horse was admitted to the Equine Hospital, Korea Racing Association with signs of colic. Based on the size of impactions, the clinical signs, the results of abdominal paracentesis and medical treatment, the prognosis was poor. The horse died 3 hours later following hopeless discharge. At necropsy, the caecum and large colon were fully filled with fecal contents and there was a rupture (10 cm in dia) in the latero- ventral caecum. The mucosa of the ileo-caecal and caeco- colic valves appeared to the hyperemic, edematous and ulcerous. There were many tapeworms in the affected mucosa. Histopathologically, lesions included hyperaemia, a deep necrotic inflammatory lesion and ulcers in the mucosa and submucosa of ileo-caecal and caeco-colic valves. One hundred thirty four faecal samples were obtained from 16 stables and submitted to parasitic examination. A total of 4 genera of eggs were recovered: Stongylus spp (82.1%), Anoplocephala perfoliata (10.5%), Bovicola equi (0.7%) and Parascaris equorum (1.5%). The major findings in this study are the presence of A perfoliata and its suspected association with the colic which led into an eventual caecal rupture. This study indicates the needs for an epidemiological survey of colic that is associated with Anoplocephala.

Animals↗

Coenurus cerebralis infection in Ethiopian highland sheep: incidence and observations on pathogenesis and clinical signs.

An investigation was carried out at Debre Berhan, Ethiopia, between 1996 and 1997, into the epidemiology of coenurosis in Menz and Horro breeds of sheep. A total of 37 heads from clinically sick and 183 heads from apparently healthy sheep were examined post mortem for the presence of the cystic larvae of Taenia multiceps, of which 37 and 5 heads, respectively, contained 1 to 8 coenurus cysts (diameter 0.8 to 6.5 cm). The bladder worms were located in the cerebral hemisphere in 96% of the cases (43% and 57% for left and right, respectively), 4% being in the cerebellum. Prediction of cyst locations based on the direction of circling and head deviation had a 62% success rate. A retrospective study generated from the health record book at the ILRI Debre Berhan Station in 1992-1996 (199 Horro and 174 Menz) revealed that the incidence of coenurosis ranged from 2.3% to 4.5%. There was no significant breed difference in the incidence. The age of the affected sheep ranged from 4 to 96 months, with a mean of 19.3 months, and 72% of the cases were within the range of 6 to 24 months. In a complementary survey on necropsied stray dogs, 8 out of 17 were positive for Taenia spp. Both studies confirm the endemicity of coenurosis at the ILRI Debre Berhan Research Station. Appropriate strategies for the control of coenurosis are suggested.

Animals↗

Tetraphyllidean plerocercoids from Western Mediterranean cetaceans and other marine mammals around the world: a comprehensive morphological analysis.

Tetraphyllidean plerocercoids have occasionally been reported in marine mammals, but they have rarely been described in detail, and the ecological significance of these infections is unclear. We described plerocercoids collected from the mucosa of the terminal colon and rectum, the anal crypts, and the hepatopancreatic ducts of 7 striped dolphins Stenella coeruleoalba, 1 Cuvier's beaked whale Ziphius cavirostris, and 3 Risso's dolphins Grampus griseus from the Spanish Mediterranean. We also examined undescribed plerocercoids from 3 cetacean species from the Atlantic and the Pacific. All plerocercoids had a lanceolate body, and a scolex with an apical sucker and 4 sessile monolocular bothridia. The bothridia had free posterior edges and an accessory sucker at their anterior end. Under light microscopy, the bothridia of some Mediterranean specimens looked bilocular without accessory suckers, but a true accessory sucker was observed in histological sections. A principal component analysis revealed 2 stable clusters of specimens along the first principal component regardless of host species. These "large" and "small" morphotypes are thought to represent early migratory stages of Phyllobothrium delphini and Monorygma grimaldii. The similarity in scolex morphology, the observation of plerocercoids buried in intestinal regions close to the sites where M. grimaldii and P. delphini occur, and the coexistence of all larval forms in the same individual hosts would support this hypothesis. Future molecular analysis may confirm it.

Anal Canal↗

Taeniacidal efficacy of SQ 21,704 in dogs by various types of oral administration and in comparison with niclosamide and bunamidine hydrochloride.

The streptothrinin antibiotics SQ 21,704 was evaluated against naturally occurring Taenia pisiformis and Dipylidium caninum infections in dogs when they were given at a dose level of 37.5 mg/kg of body weight in four different rations: loaf-type canned meat; chunk-type canned meat; dry (gravy-type) meal; and dry (pelleted) meal. The SQ 21,704 was 100% efficacious against both T pisiformis and D caninum infections when given with the chunk, gravy, and pelleted rations. When given with the loaf-type canned meat, it was 100% effective against T pisiformis and 60% efficacious against D caninum. The SQ 21,704 was effective against both tapeworm species when given orally as a liquid at a dose level of 37.5 mg/kg, formulated as an aqueous suspension containing 94 mg of activity per milliter. The SQ 21,704 was also tested in dogs when given orally in gelatin capsules at a dose level of 37.5 mg/kg without fasting, and was 100% efficacious against both tapeworm species. The results of a comparative taeniacidal study demonstrated that SQ 21,704 was 100% effective in removing both T pisiformis and D caninum when administered orally at a dose level of 37.5 mg/kg, whereas niclosamide and bunamidine hydrochloride were only partially effective at their recommended dose levels. One of five dogs treated with niclosamide at a dose level of 157 mg/kg was positive at necropsy, giving an orally efficacy of 80%. Three of five dogs treated with bunamidine hydrochloride at a dose level of 49 mg/kg were positive at necropsy, giving an overall efficacy of 40%.

Administration, Oral↗

A comparison of mode of attachment and histopathogenicity of four tapeworm species representing two orders infecting the spiral intestine of the nurse shark, Ginglymostoma cirratum.

This study was undertaken to compare 2 species of Tetraphyllidea and 2 species of Trypanorhyncha with regard to the relationship between attachment structure morphology, mode of attachment, and tapeworm size, to damage at the sites of attachment in the Atlantic nurse shark, Ginglymostoma cirratum. Regions of the spiral intestine with worms attached were removed from 8 nurse sharks and sectioned according to conventional techniques. Sections of 5-50 specimens of each tapeworm species were examined. Regions of the spiral intestine devoid of worms were processed for characterization of the normal mucosa. The normal mucosa was found to consist of a folded surface covered with round-to-oval primary mucosal crypts. In the first 7 or 8 chambers of the spiral intestine the mucosal surface was thrown into secondary folds, forming ridges and secondary crypts. The primary mucosal crypts were lined with a single layer of columnar epithelium resting on a basement membrane. A highly cellular lamina propria and submucosa were found between the crypts and the muscularis mucosa. The small tetraphyllidean Pedibothrium brevispine was found with its scolex lying within the primary mucosal crypts with its hooks embedded in the basement membrane. Epithelial denudation was evident. The large tetraphyllidean Pedibothrium globicephalum was found with its bothridia engulfing large portions of the mucosa and its hooks embedded into the lamina propria. It was associated with moderate to severe mucosal necrosis. The small trypanorhynch Prochristianella tenuispine was found lying between the mucosal ridges in the secondary crypts with its tentacles either penetrating the epithelium, or occasionally, the lamina propria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of a Gyrodactylus salaris variant: infection biology, morphology and molecular genetics.

A laboratory population of a Danish Gyrodactylus salaris variant founded by 1 single specimen was established and infection studies performed. Rainbow trout as well as Atlantic salmon of 3 different stocks were infected both in cohabitation systems and as single-parasite infections on isolated hosts. Both infection systems revealed that this particular morphotype exhibits low virulence towards Atlantic salmon. Thus, in isolated hosts, the parasites could either not establish or only reproduce to a limited degree on salmon. Rainbow trout, in contrast, proved to be rather susceptible to infection with this G. salaris variant and abundances on this host species always attained significantly higher values in cohabitation systems compared to salmon. Detailed morphological examination confirmed the very high resemblance to G. salaris (sensu stricto), as the range of variation in sclerite characters of the morphotype is almost fully covered by the total range of variation reported for reference G. salaris. Morphological similarities to the closely related congeneric species G. bohemicus were noted. Molecular studies, however, showed that the morphotype most likely represents a G. salaris variant, as it differed only slightly from G. salaris sensu Malmberg, 1957, which is also known to inhabit Danish watercourses. The genomic target region investigated does not allow us to rule out the possibility that it represents a variant form of G. thymalli. Sequences of the ribosomal RNA internal transcribed spacer (ITS) revealed that single individuals contained 2 different ITS sequences, one identical to reference sequence of G. salaris while the other differed at 3 positions. This can be interpreted as an example of a hybrid or, more likely, as intra-individual variation of ITS within single individuals. As one of the nucleotide changes in the variant ITS affects an Hae III restriction site, the current G. salaris variant can be distinguished from G. salaris sensu Malmberg by RFLP diagnosis.

Animals↗

Amplification of ribosomal DNA of Anoplocephalidae: Anoplocephala perfoliata diagnosis by PCR as a possible alternative to coprological methods.

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.

Animals↗

Preparation and surface labeling of murine eosinophils.

Eosinophilic polymorphonuclear leukocytes were isolated from the peritoneal cavity of BALB/c mice infected with the parasite Mesocestoides corti. Approximately 4 X 10(7) eosinophils (purity, 50%) could be harvested from each mouse. A high yield and purity of eosinophils was obtained from the peritoneal cells of infected male BALB/c mice using density centrifugation on a gradient of slightly hypotonic colloidal silica sol (Percoll). After initial irradiation of the mice to lower the lymphocyte contamination, subsequent density gradient (and where necessary sedimentation velocity) centrifugation yielded 10(8) eosinophils (purity > 95%) from six to eight mice. It was also possible to isolate small numbers of eosinophils (2 X 10(4) cells/minute, purity > 99%) without irradiating the mice. This could be achieved by separating the density gradient purified peritoneal cells by light-scatter on a Becton-Dickinson cell sorter (FACS II). Analysis of proteins extracted from eosinophils using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed a group of high molecular weight proteins (bwtween 250K and 160K) which were not as distinctive in the neutrophil profile. Surface labeling was performed, before the cell separation by using 125I and 1,3,4,6-tetrachloro-3 alpha, 6 alpha-diphenylglycoluril. Only five 125I-labeled proteins were detected initially (all with apparent molecular weights > 50,000). No 125I appeared to be associated with actin under the conditions used for surface labeling. Four of the eosinophil surface labeled proteins corresponded to surface labeled proteins on neutrophils, but the major surface component of the eosinophils (MW 79,000) appeared to be smaller than the major neutrophil protein (MW 90,000).

Animals↗