Demonstration of Isospora suis oocysts in faecal samples.
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Biomedical subjects
Publications and source records attributed to S A Henriksen.
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Immunity to Ostertagia ostertagi infections in calves develops slowly and only becomes manifest towards the end of a grazing season in which they have been exposed to the parasite. In an attempt to hasten the onset of immune reactions, three immunization protocols were set up. Twenty four heifers were allocated into four groups. Beginning in January, animals in two of the groups were inoculated with four 1-monthly increasing dosages of either 'normal' or 'chilled' (hypobiosis-prone) larvae, those in the third group received a single large infection with 'chilled' larvae and those in the fourth group served as non-infected controls. All animals were turned out on a common pasture in late April. Development of immunity was evaluated through determinations of faecal egg counts, live weight gains, serum pepsinogen levels and specific serum antibody responses of three isotypes (IgG1, IgG2 and IgA). Significantly reduced egg excretions in the immunized groups were apparent early in the season, indicating that the immunizations had, in this respect, been efficacious. The 'chilled' and 'normal' larvae seemed equally efficient given as multiple and single infections. A single large dosage of 'chilled' larvae seemed to have adverse effects. Only moderate antibody responses were elicited probably because of low challenge infection level on pasture. Considerable variation in responses existed between and within the four groups, for which reason conclusions regarding correlations between antibody isotype responses and immune effects on parasites could not be made.
The experiment was designed to test the survival and performance of stress selected nematophagous fungi after passage through the gastro-intestinal tract of cattle. Ruminating calves were fed daily with a fixed amount of fungal material grown on barley grains. The excreted dung was collected on days four and five after the start of the feeding experiment. Barley grains were washed out of the excreted dung and incoculated on water-agar plates. After incubation for one week, nine of ten fungal isolates were re-isolated from these plates. The predatory capacity of the fungi in the excreted faeces was tested in a dung pat bioassay and a faecal culture system. In the dung pat bioassay, two fungi of the genus Arthrobotrys and six of the genus Duddingtonia reduced the development of Ostertagia ostertagi third stage larvae by 85% (61%-93%), compared to the number of larvae developed from fungus-free control pats. In seven out of these eight isolates, the reduction of larvae in the faecal cultures was 92% (76%-99%).
A study on the histamine release test (HR) for the demonstration of infections with Trichinella spiralis in pigs was carried out on 18 pigs, six infected with 200 larvae, six infected with 5000 larvae and six non-infected (control group). The results obtained by HR during a 7 week infection were compared with those of the enzyme-linked immunosorbent assay (ELISA). All inoculated pigs were found to be positive on Day 40 post-inoculation (p.i.) by necropsy examination of selected muscle groups, with mean recoveries of 7.9 and 225 larvae g-1 of tissue in the low- and high-dose group, respectively. At this time, all animals of the high-dose group and five out of six animals of the low-dose group were antibody positive in ELISA with any of three coating antigens employed (a crude muscle larva extract, an excretory/secretory (ES) antigen and a purified 45 kDa antigen). HR performed on whole blood was positive in four out of six pigs of the high-dose group and one out of six pigs of the low-dose group. The earliest ELISA seroconversions took place at Day 15 p.i. with crude and ES antigens. The earliest measurable reaction in HR performed on whole blood was found on Day 19 p.i. There was considerable individual variation regarding which test was the most sensitive for the early detection of infection. Washing of the blood cells prior to antigen provocation led to a markedly improved sensitivity of HR, all animals of the high-dose and three out of six animals of the low-dose group being positive by Day 40 p.i. The time course of the development of ELISA titres and HR reactivity indicated that this effect is due to the removal of blocking antibodies.
Laboratory experiments were designed to select nematophagous fungi that were able to survive in vitro conditions simulating passage through the gastro-intestinal tract of cattle. All of the tests were conducted at 39 degrees C. In a primary stress selection step in diluted rumen fluid, 21 isolates were obtained. Each of the primary stress selected isolates was tested in synthetic saliva, rumen fluid simulating the activity in the rumen, rumen fluid followed by pepsin-hydrochloric acid treatment simulating the additional effect of ruminal and abomasal activity, pepsin-hydrochloric acid solution simulating conditions in the abomasum and finally in a trypsin solution as an example of enzyme activity in the gut. The effect of the rumen fluid alone, or rumen fluid followed by pepsin-hydrochloric acid treatment, were responsible for the reduction in surviving fungal isolates. Only six of thirteen isolates belonging to the genus Arthrobotrys survived while seven of eight isolates of the genus Duddingtonia survived. Fourteen isolates were tested for their predatory capacity in a dung pat bioassay. Fungi of the genera Arthrobotrys and Duddingtonia reduced the development of Ostertagia ostertagi third stage larvae by approximately 75% and 96% respectively compared to the number of larvae that developed from fungus-free control pats.
Serum IgG response of cattle with cysticercosis caused by Taenia saginata was studied in an enzyme-linked immunosorbent assay (ELISA) where a T. saginata metacestode surface extract was used as antigen. In experimentally infected calves, a sharp rise in specific antibody levels was found 3-4 weeks after the infection followed by a logical level of detection corresponded to about 25 cysts. The ELISA was employed in cattle herds where cysticercosis outbreaks had occurred and also in supposedly uninfected herds. Significantly increased antibody levels were found in the herds with massive cysticercosis cases. The test was not adapted for individual diagnosis as some animals of the uninfected herds, especially within the older age groups, had elevated antibody values. The ELISA was, however, useful in the investigation of outbreaks to determine the extent and pattern of the infection in the herd. The rate of decline in antibody levels in these herds was studied by follow up sampling. The increased antibody levels in the infected herds were also reflected in colostrum-fed calves. This observation was employed to estimate the time of infection.
Risk factors in light T. saginata cysticercosis in cattle herds were investigated in a case-control study. The case group consisted of farms from which cattle with low-grade cysticercosis had been detected at slaughter. The control group consisted of farms where no cases were detected during the same period. The major risk factor identified was allowing cattle access to drink from streams carrying effluent from sewage treatment plants (odds ratio = 3.6). Spreading of septic tank sludge through deliberate or accidental mixing with animal slurry, deposition of sewage sludge, and proximity to railways, camping sites or sewage treatment plants seemed to be of minor importance.
This study was undertaken to establish whether anthelmintic resistance was present in nematode parasites of horses in Denmark. Sixteen horse farms were selected for faecal egg count reduction (FECR) tests to measure the efficacy of the anthelmintic used. Resistance to benzimidazole anthelmintics was found on 13 of the 16 farms, with FECR values ranging from 80.0% to -101.3%. On the remaining 3 farms FECR was 100.0%, 99.3% and 97.2%. Results of a questionnaire study on anthelmintic usage, parasite control measures and management practices showed that horses in this study were treated on average 7.1 times/year. Horse owners changed between preparations of drugs but almost only within the same class of anthelmintics. Nine owners gave an anthelmintic treatment to purchased horses before they were introduced on the farm. On 14 farms, the same paddock was grazed every year and the average stocking rate was estimated to be 2.4 horses/ha. Strategies to avoid development of anthelmintic resistance are discussed and recommendations of parasite control on horse farms are presented.
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The syringe and the modified Bell filtration techniques, the cellophane faecal thick smear (Kato/Katz) and Teesdale (sandwich) smear techniques, and the formol-ether concentration technique were compared for efficiency for detection of eggs of Schistosoma bovis in faeces from cattle and goats. Both filtration techniques were highly effective as revealed by high sensitivity and mean egg counts. However, efficiency declined markedly using formalin-fixed faecal material. The efficiency of the Teesdale technique was comparable to that of the filtration techniques, whereas the efficiency of the Kato/Katz technique in goats and the formol-ether concentration technique in cattle was markedly lower. The ease of performance combined with the high sensitivity make the syringe filtration and the Teesdale techniques recommendable for field diagnosis of schistosomiasis in domestic stock.
This study on natural trichostrongylosis in calves on permanent pasture was designed to evaluate whether susceptible first-season calves would benefit from mixed grazing with second-season calves that were more resistant. The experimental plan allowed a comparison of the effect of mixed grazing with that of separate grazing for each age category. The results showed that the first-season calves already grazing with the older animals at the beginning of the grazing season had lower parasite burdens and less clinical symptoms than first-season calves grazing alone. This effect, experienced in 1987 and characterized by a rather high overwintered parasite population on herbage, could not at this early stage be explained by a parasite "diluting" effect exerted by the older animals, but might perhaps be explained by different grazing behaviour of the two age categories. Later in the summer, the first-season calves continued to benefit from the mixed grazing, most likely because the pasture was less contaminated per unit area due to the very low egg deposition of the older animals. The effect of mixed grazing was most clearly reflected in the differences between worm burdens in tracer calves grazing the respective plots at the end of the season. Apparently, the second-season calves did not suffer parasitologically from grazing together with the younger animals. The paper discusses possible long-term implications with combinations of different age categories of cattle.
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The infectivity of Taenia saginata eggs exposed to environmental conditions on a natural soil surface in Denmark was studied by feeding the eggs to susceptible calves, followed by determination of the number of cysts developed. The results indicated that a small proportion of the eggs remained infective for 6 1/2 months, but not for 9 1/2 months when deposited in May 1986, and for 5 1/2 months but not for 8 1/2 months when deposited in September 1987. Viability of eggs was tested in vitro and compared with infectivity obtained in calves.
Epidemiological studies were made on 14 farms from which, during a 2 year period, 38 cattle had been condemned at slaughter, due to massive infections with Cysticercus bovis. By on-site investigations and interviews, attempts were made to identify the transmission routes of Taenia saginata eggs from human faeces into the environment and further on to cattle. The most frequent sources of infection were found to be sludge from septic tanks illegally applied on pasture or crops, in some cases after having been mixed with animal slurry. Animals in permanently housed herds were infected through the fodder or by contamination of the indoor environment by such slurry containing Taenia eggs. Other herds were infected by grazing pastures in close proximity to municipal sewage treatment plants. In contrast to earlier Danish observations, application on farmland of sewage sludge from municipal treatment plants was not involved in any of the reported outbreaks. This apparent change coincides with the implementation of more restrictive legislation for the agricultural use of sewage sludge in Denmark.
The distribution of Taenia saginata cysts was recorded in 23 experimentally infected calves. The calves harboured between two and 2569 cysts. A median of 15.7 per cent of the cysts were located in the heart and 6.5 per cent in the masseter muscles, the organs usually considered as the most important predilection sites. From these findings a simple probability model was used to estimate the meat inspection sensitivity at different infection levels. It was concluded that the possibility of detecting infected cattle is limited especially when, as in Denmark, the majority of animals are lightly infected.
This paper describes clinically manifest infections occurring as early as June and early July in first-season grazing calves in Denmark during the period 1972-1987. Two severe outbreaks in 1976, included in our experiments, were studied in detail. Herbage infectivity was particularly high in grass tufts surrounding cow pats that were present in high numbers around the time of turnout. It is hypothesized that the preceding extremely dry summer followed by a hard winter had indirectly retarded degradation of dung pats and thereby favoured the overwintering of the larval populations in the dung reservoirs. In a third experiment, conducted in the same year, the epidemiological pattern was more normal; presumably due to artificial irrigation of the pasture during the drought. Early-season cases that were recorded in 1980 and 1987 could possibly be related to cold winters and/or very early turnout. The findings are discussed in the light of ecological factors responsible for the breakdown of cow pats. The clinical implications are seen in relation to current control methods.
The larval recovery rates of Ostertagia ostertagi and Cooperia oncophora from monospecific copro-cultures were determined by the methods of Corticelli and Lai, and Henriksen and Korsholm using vermiculite or polystyrene pellets as inert additives. The addition of either vermiculite or polystyrene pellets in the two culture methods significantly (P less than 0.05) improved larval recoveries of both parasite species. Recovery rates with the Henriksen and Korsholm method were higher (P less than 0.05) for both species than with the Corticelli and Lai method. Advantages and disadvantages inherent in the use of the two culture methods with and without additives are discussed.
Artificially prepared cow pats containing Ostertagia ostertagi eggs were deposited on two comparable grazing plots at weekly intervals during June and July 1987. Before deposition the cow pats, on the one plot, were inoculated with 0.250 g mycelial fragments per kg of faeces of the predacious fungus Arthrobotrys oligospora. On the other control plot comparable numbers of non-inoculated cow pats were placed at the same time. Subsequently, in July, two equal groups of calves were turned out one on each plot to graze for two months after which they were necropsied. The number of infective O. ostertagi larvae in inoculated cow pats was reduced by 42% and herbage larval infectivity around them by 50-71% as compared with the corresponding parameters in control pats and surrounding herbage. After a grazing period of two months the calves on the plot with inoculated cow pats had acquired worm burdens that were on average 37% lower than those of calves on the control plot. The lower degree of parasitism in these calves was also reflected in lower egg counts, lower serum pepsinogen levels and higher body weight.