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Biomedical subjects

J Eckert

Publications and source records attributed to J Eckert.

At least 145 records · Page 8Linked to original sources

Cryopreservation of parasites.

In this review, advances in cryopreservation of helminth parasites are reported. Our own studies demonstrate that metacestodes of Echinococcus multilocularis can be maintained in a viable state for at least 1-2 years by appropriate deep-freezing and storage in liquid nitrogen. Infective larvae of the nematode Toxocara canis cryopreserved for 1 week in liquid nitrogen were maintained after thawing in vitro in a chemically defined medium for 35 weeks. Although motility of previously deep-frozen larvae was reduced they produced secretory/excretory antigens of similar immunodiagnostic quality as those from unfrozen larvae. Whereas infective larvae of several species of trichostrongy-lids can be easily cryopreserved, the infective larvae of the cattle lungworm, Dictyocaulus viviparus, and muscle larvae of Trichinella spiralis are more sensitive to damage by subzero temperatures. Therefore, survival rates after cryopreservation are low, but improvement of the cooling schedules appears to be feasible. It is concluded that cryopreservation of certain stages of helminth and protozoan parasites is a useful technique for long-term storage of defined isolates, which can contribute considerably to reducing the number of experimental animals usually required for serial passages.

Animal Testing Alternatives↗

Echinococcus strains in Europe: a review.

In European countries the following final/intermediate host assemblages of Echinococcus granulosus are known to occur: (a) dog/sheep; (b) dog/horse; (c) dog/cattle; (d) dog/pig, (e) dog/reindeer (= "northern form" of E. granulosus). The parasites involved in cycles a-d have been identified as distinct strains in various countries. Epidemiological aspects relevant to intraspecific variations and strain characteristics are described. In E. multilocularis there are some indications for intraspecific variability but it is yet unknown whether or not they are of epidemiological or other significance. There is clear evidence that E. multilocularis, the dog/sheep strain and the "northern form" of E. granulosus are infective for humans; the status of the other strains regarding infectivity to man is still unclear.

Animals↗

Cryopreservation of Dictyocaulus viviparus third-stage larvae and Trichinella spiralis muscle larvae.

In cryopreservation studies with third-stage larvae of Dictyocaulus viviparus, best results were achieved by incubating larvae in 0.05% NaOCl at 37 degrees C to remove the sheath, followed by cooling at a rate of 1 degree C min per min down to about 0 degree C. After an equilibration time of 10 min at +4 degrees C with or without 4% polyethylene glycol-400 as cryoprotectant, samples were frozen at the same cooling rate to an intermediate temperature of -20 degrees C, maintained at this temperature for 10 min and finally plunged into liquid nitrogen for storage. Three groups of 3 calves were infected with the following batches of third-stage larvae: (a) fresh, sheated; (b) fresh, exsheathed; (c) exsheathed, cryopreserved for 13 weeks in liquid nitrogen and subsequently thawed. Although 62% of group (c) were regarded as viable in vitro, their infectivity to calves was low and only an average of 0.08% of the inoculated larvae (3000 per animal) developed into adult lungworms (= infectivity rate). Average infectivity rates of fresh, sheathed (a) and fresh, exsheathed (b) larvae were much higher (38.3% and 29.7%) and not significantly different from each other. Two of the calves inoculated with previously frozen larvae and all of the calves infected with fresh larvae excreted first-stage larvae in their faeces, but the latter groups in higher quantities. The results show that cryopreservation of exsheathed third-stage larvae of D. viviparus is possible, but for strain maintenance infection doses greater than 3000 larvae should be used for inoculation of calves.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Infectivity of Swiss Giardia isolates to jirds and mice, and in vitro cultivation of trophozoites originating from sheep.

The susceptibility of Giardia-free jirds (Meriones unguiculatus) and AJ-strain mice to oral inoculation with cysts from Swiss Giardia isolates was assessed. Giardia cysts of human, dog, cat, and sheep origin produced infections in jirds (4-10 weeks old). AJ-strain mice (6 weeks old) were insusceptible to Giardia from man, dog, and cat. The number of Giardia cysts inoculated per rodent ranged from 5000 to 12,600. The course of infection was studied in jirds experimentally infected with a Giardia isolate of cat origin. After administration of 6300 cysts to each of 16 jirds, 5 (31%) started to excrete cysts 4-5 days post inoculation (p.i.). Cyst excretion patterns were mostly intermittent, patency periods lasted 5-28 days, and average cyst numbers per g feces of individual jirds varied between 118,500 and 835,200 (maximal number: 3216700). The parasites were eliminated spontaneously in all 5 jirds. The Giardia isolate of cat origin was passaged 4 times in the jird. A Giardia isolate obtained from sheep was transferred in the cyst stage to jirds. Giardia trophozoites isolated from one of these jirds have been axenically cultivated in vitro in a modified, filter-sterilized TYI-S-33 culture medium and successfully cryopreserved. To our knowledge, this is the first report on axenic cultivation of Giardia of sheep origin. Giardia from cats could not be cultivated in vitro under the same conditions. The fact that Swiss Giardia isolates from humans and various domestic animals are not strictly host-specific is in agreement with similar findings from other areas and suggests a zoonotic character for giardiasis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cryopreservation and long-term in vitro maintenance of second-stage larvae of Toxocara canis.

Second stage larvae of Toxocara canis were isolated from developed eggs, frozen in Eagle's Minimal Essential Medium with 5% dimethyl sulfoxide or 10% glycerol as cryoprotectants according to two cooling schedules and maintained in liquid nitrogen for 1 week. After thawing, the previously frozen larvae (FL) and unfrozen controls (CL) were maintained in a chemically defined medium in vitro for 35 weeks. While CL had motility rates around 95% to 97% throughout the experiment, previously frozen larvae (FL) exhibited rates of 48%-58% at the beginning and of 19%-39% at the end of the 35 week in vitro maintenance period. The surviving FL and CL larvae proved to be infective for mice. Excretory/secretory (ES) antigens isolated from several batches of culture medium in which FL and CL had been maintained reacted in the ELISA with human sera containing antibodies against Toxocara. Antigens from FL and CL separated by SDS-PAGE and silver-stained showed some differences in polypeptide patterns. Western-blot analysis revealed that these differences were not related to antigenic polypeptides but were most likely caused by substances without antigenic properties originating from dead and/or degenerating larvae. It can be concluded that ES antigens produced by previously frozen larvae are essentially the same as those derived from unfrozen controls. The value of cryopreservation of T. canis larvae for routine production of ES antigens will be further evaluated.

Animals↗

Sero-epidemiological survey for alveolar echinococcosis (by Em2-ELISA) of blood donors in an endemic area of Switzerland.

Sera from 17,166 blood donors living in 10 cantons of northern Switzerland in an area endemic for Echinococcus multilocularis were investigated by serological survey for alveolar echinococcosis (AE). A highly species-specific antigen (Em2) and a commonly used E. granulosus hydatid fluid antigen (EgHF) were compared for their suitability in seroepidemiology. EgHF showed a degree of nonspecificity which did not allow direct detection of AE cases. Antibody reaction with Em2 resulted in the detection of 2 asymptomatic clinical cases of AE (seroprevalence 0.01%) within this population of blood donors. A further 4 persons were positive in Em2-ELISA. These 4 persons had negative imaging studies and will be followed serologically and clinically.

Adult↗

[New findings on echinococcosis].

The following new aspects of cystic and alveolar echinococcosis (= infections with the metacestode stages of Echinococcus granulosus and E. multilocularis respectively) are reported: identification of a Swiss E. granulosus isolate as "cattle strain" which differs from the "sheep strain"; new observations on proliferation and metastasis formation of larval E. multilocularis; information on chemotherapy of human echinococcosis; recent developments in immunoserology. The latter includes a new technique for serological differential diagnosis of cystic and alveolar echinococcosis, the determination of parasite-specific immunoglobulin classes (IgG, IgM, IgA and IgE) and circulating antigens in ELISA, and the introduction of arc-5 detection in routine serodiagnosis. A highly purified, species-specific antigen from E. multilocularis is now available for seroepidemiological studies.

Anthelmintics↗