Cross-sectional serological survey on gastrointestinal and lung nematode infections in first and second-year replacement stock in the netherlands: relation with management practices and use of anthelmintics.

A cross-sectional survey was carried out on 86 farms randomly distributed in The Netherlands. After housing following the first and the second grazing season (FGS and SGS) serum samples were collected to determine IgG levels against Cooperia oncophora and Dictyocaulus viviparus, and the pepsinogen content. A questionnaire was used to inquire on grazing management practices and the use of anthelmintic drugs. On 80.7 and 60.2% of the farms FGS and SGS animals, respectively, were treated at least once with an anthelmintic drug. The percentage for the SGS animals indicates that the use of anthelmintic drugs in those animals has increased enormously over the last 10-15 years. Generally, parasitic nematode control in the FGS is good on most farms, but it can be characterised as being overprotective. There is a tendency that if anthelmintic drugs are used in the FGS they also are used more often in the SGS. On 12 farms (14%), no anthelmintic drugs were given in the FGS and the SGS. These farms did not differ from the others with respect to management practices in any obvious way. The serological results were in general very low, indicating low levels of exposure to gastrointestinal nematode infection in both FGS and SGS animals. This was not surprising in view of the good to high level of nematode control practices reported by the farmers. Although not statistically significant, a consistent result was that serological results for the SGS animals were more often positive or on average higher on those farms where FGS parasite control tended to be excessive. For D. viviparus, a prevalence rate of 41% positive farms was found. Following comparison with previous data, it is speculated that lungworm (sero-)prevalence in replacement stock may be declining as a result of continuing high levels of parasite control in replacement stock. It is concluded that the results confirm previous surveys, lending support to the conclusion that parasitic nematode control on Dutch dairy farms, certainly in FGS calves, is good but tends to be overprotective.

Cultured pig rhabdomyosarcoma cells with a deletion of the Xq24-qter chromosome region: an immunochemical and cytogenetic characterization.

A pig rhabdomyosarcoma cell line (PRUM59) was established, and the immuno(histo)chemical and cytogenetic characterization of these cells was determined. At various swine farms in the Netherlands, pigs were observed that had solitary or multiple skin nodules, which were diagnosed as rhabdomyosarcomas. Cells of a tumor derived from a 3.5-week-old female pig were cultured for immunochemical and cytogenetic analyses. The cell line had characteristic features of undifferentiated muscle cells, similar to those observed in tumor tissue sections; they contained titin, a high-molecular weight protein specific for striated muscle, as dot-like aggregates and as filaments, desmin filaments and cross-striations, smooth muscle actin stress fibers, and vimentin filaments. The cells stained positively for striated muscle actin and tropomyosin as well. The immunohistochemical staining results were supported by results of immunoblotting experiments. Karyotyping of the cells revealed a deletion of a major part of Xq24-qter, a part of the long arm of 1 of the 2 X chromosomes. The other X chromosome and all autosomes appeared to be normal.

Optimization of the PCR test for the mutation causing bovine leukocyte adhesion deficiency.

The recent emergence of the bovine leukocyte adhesion deficiency (BLAD) demonstrated the risks of narrowing the genetic basis of a population. About 6% of the Holstein-Friesian cattle now descends from one bull who was a heterozygous BLAD carrier. Crossing his descendants resulted in the birth of homozygous BLAD calves with a life expectancy of < 1 year. The BLAD syndrome is caused by a point mutation in the gene coding for CD18, a subunit of the beta 2 integrins on the surface of leukocytes. By using a PCR-RFLP test, large numbers of cattle are now being screened in several countries to eradicate the mutant allele. We describe an optimization of the PCR primer set that has led to an improvement of the test.

[Changes in the histochemical composition of mucins in goblet cells in the course of a Nippostrongylus brasiliensis infection in the rat]. / Veranderingen in de histochemische samenstelling van mucinen in slijmbekercellen tijdens het verloop van een infectie met Nippostrongylus brasiliensis in de rat.

Rats were infected with Nippostrongylus brasiliensis, and changes in the histochemical composition of mucins in jejunal goblet cells were investigated. Ten days after infection, mitotic activity was extensively in jejunal crypts: both crypts and villi were characterised by hyperplasia of goblet cells. Infected rats had a markedly greater number of crypt and villi goblet cells containing neutral mucin than the control rats did. Moreover, 15 days after infection, infected rats had significantly more goblet cells containing acid mucin than control rats did. In infected rats, the acid mucins in goblet cells (day 15) appeared to be predominantly sulphomucins, whereas in control rats the acid mucins were predominantly sialomucins. The experiments established that when N. brasiliensis is excreted by rats, the histochemical composition of mucins in crypt and villi goblet cells has been changed not only quantitatively, but also qualitatively.

Nippostrongylus brasiliensis: histochemical changes in the composition of mucins in goblet cells during infection in rats.

Changes in the quality of mucins in jejunal goblet cells were investigated during an infection with Nippostrongylus brasiliensis in rats. At 10 days after infection, when proliferative activity in the crypts is excessive and both crypts and villi are characterized by hyperplasia of goblet cells, the histochemical composition of the population of goblet cells in comparison with controls shows a marked increase in crypt and villous goblet cells containing neutral mucins. At 15 days after infection both crypts and villi display a significant increase in goblet cells containing acid mucin and decrease in goblet cells containing neutral mucin. The acid mucins in crypt and villous goblet cells on day 15 appear to be sulphomucins predominantly, whereas in controls sialomucin-containing goblet cells dominate both in the crypts and on the villi. These experiments establish that the explusion of N. brasiliensis from the intestine of the rat coincides not only with quantitative, but also with remarkable qualitative changes in the histochemical composition of mucins in goblet cells.

Histochemical changes in the composition of mucins in goblet cells during methotrexate-induced mucosal atrophy in rats.

Changes in the composition of mucins in jejunal goblet cells were investigated histochemically after a single intraperitoneal injection of methotrexate in rats (40 mg methotrexate/kg body weight). At 48 h after injection of methotrexate, when mitotic activity in the crypts approximates to zero, an almost complete disappearance of mucin-containing goblet cells can be observed. At 96h, when proliferative activity in crypts is excessive, neutral mucin- and sialomucin-containing goblet cells dominate in the crypts, whereas in controls the crypts display sulphomucin-containing goblet cells. At 144h neutral mucin- and sulphomucin-containing goblet cells dominate in the crypts. Both at 48 and 96h after injection of methotrexate a significant decrease in the number of villous goblet cells takes place, whereas with respect to the quality of the mucins in the goblet cells no difference can be found in comparison with control rats. At 144h the villi have recovered from the preceding temporary decrease of goblet cells. The histochemical composition of the population of goblet cells is still aberrant and shows goblet cells containing neutral mucins and sulphomucins principally, whereas in controls sialomucin-containing goblet cells dominate on the villi. The established changes in the composition of mucins in goblet cells cannot be attributed to methotrexate-induced malnutrition, as pair-fed control rats clearly demonstrate. Our results clearly demonstrate that conspicuous quantitative as well as qualitative changes in the synthesis of mucins in goblet cells occur during the hyporegenerative and hyperregenerative phase of methotrexate-induced mucosal atrophy.

Comparison of two control systems for cyathostome infections in the horse and further aspects of the epidemiology of these infections.

The small strongylid infections of two groups of three yearling female Shetland ponies and one yearling Shetland tracer pony were studied. One group was set stocked from April to November and was treated monthly with 5 mg kg-1 albendazole from two days before turnout until July. The other group grazed similar pasture until July, was treated with 5 mg kg-1 albendazole and subsequently removed to pasture grazed by sheep from April to July. The tracer ponies were added to both groups in September. The efficacy of both methods was not completely satisfactory probably because of low efficacy of anthelmintic treatment. There were no significant differences between the cyathostome burdens of the two groups. A high proportion of the cyathostome populations of all ponies consisted of inhibited early third stage larvae (L3). The finding of low numbers of immature fifth-stage worms in the tracer ponies indicated that the considerable adult burdens in the permanent ponies originated from infection picked up before the tracer ponies were added. In the group which was removed to sheep pasture after treatment in July it was likely that the majority of the adult worm burden had been ingested as infective larvae before treatment.

The distribution of inhibited early third stage Cyathostominae larvae in the large intestine of the horse.

The distribution of inhibited early third stage Cyathostominae larvae in different parts of the large intestine of the horse was studied in 20 Shetland ponies necropsied in autumn 1982, 1983 and 1984. The location of the larvae in the large intestinal wall was studied by histological examination of the intestines of the eight ponies from 1984. Inhibited larvae were located predominantly and more or less equally in the caecum and the ventral colon. Generally fewer early L3 were in the dorsal colon. In 1984 a considerable proportion (mean 17%, range 9.7-36.9%) of the inhibited larvae was found in the contents instead of in the mucosa of the large intestine, despite a housing period under helminth free conditions of 5 weeks. These larvae probably had been overlooked in 1982 and 1983. In sections cut from the intestinal wall most early L3 were found in the lamina propria. They were surrounded by a small fibrous capsule. Some were found in the lumen or epithelium of the Lieberkühn's crypts and a small proportion in the submucosa.

Control of strongylosis in horses by alternate grazing of horses and sheep and some other aspects of the epidemiology of Strongylidae infections.

Alternate grazing of horses and sheep as a control measure for gastrointestinal helminthiasis was studied in three grazing experiments in 1981, 1982 and 1983. Each year a group of three mare yearling Shetland ponies, which were kept on a small pasture from spring to autumn, were compared with a similar group which grazed a similar or the same pasture until July and were subsequently removed to a similar pasture which had been grazed by sheep from April to July. In addition both groups were treated with an anthelmintic when the latter group was removed to the sheep pasture. Pasture larval counts and worm counts and, in 1982 and 1983, faecal egg counts, clinical condition, total protein, albumin and beta-globulin levels demonstrated that the groups removed to sheep pasture acquired considerably lower burdens of nematodes of the subfamilies Cyathostominae and Strongylinae, but considerably higher burdens of Trichostrongylus axei than the groups which were not moved. These T. axei infections resulted in higher serum pepsinogen levels in the former groups compared to the latter in 1981 and 1982. At necropsy an important part of the T. axei burdens and, in 1982 and 1983, the Cyathostominae burdens consisted of inhibited early third stage larvae. A total of 20 species of the subfamily Cyathostominae and 7 species of the Strongylinae were found. Generally the composition of species was in agreement with other observations in western Europe, the most common species being: Cylicostephanus longibursatus, Cylicostephanus minutus, Cylicostephanus calicatus, Cylicostephanus goldi, Cylicostephanus poculatus, Cyathostomum labratum, Cyathostomum coronatum, Cyathostomum catinatum, Cylicocyclus leptostomus, Cylicocyclus nassatus, Cylicocyclus insigne, Strongylus edentatus and Strongylus vulgaris.

Spinal nematodosis in a donkey with posterior paresis.

Spinal nematodosis in a four-year-old donkey is described. The animal showed progressive hindlimb paresis of sudden onset. Necropsy revealed extensive degeneration and inflammation in the lumbosacral part of the spinal cord, caused by a nematode larva of the suborder Strongylina, probably L4 or L5 of Strongylus vulgaris.

Inhibited development of cyathostominae in the horse in the early third stage.

Inhibited early third stage larvae of Cyathostominae were found in the digested mucosa of the large intestine of 12 yearling, female Shetland ponies, which were used in two grazing experiments. The ponies were killed in late autumn. In some ponies the inhibited larvae were very abundant and in most animals the majority of the Cyathostominae populations consisted of these early third stage larvae, suggesting that the phenomenon has an epidemiological significance.

Alternate grazing of horses and sheep as control for gastro-intestinal helminthiasis in horses.

Gastro-intestinal helminth infections of a group of three yearling mare Shetland ponies, which were set-stocked on a small pasture from February until September, were compared with those of a similar group of ponies, which grazed a similar pasture from February to July and subsequently was removed to a pasture which had been grazed by sheep from April to July. In addition both groups were treated with cambendazole when the latter group was removed to the sheep pasture. Pasture larval counts and worm counts demonstrated that the group grazed after the sheep acquired considerably smaller burdens of nematodes of the subfamilies Cyathostominae and Strongylinae, but considerably higher burdens of Trichostrongylus axei than the group which remained on the same pasture. The T. axei infections in the former group caused an increase in the serum pepsinogen levels within two weeks after removal to the sheep pasture followed by a gradual decrease. At necropsy T. axei populations consisted almost exclusively of inhibited early third stage larvae.

[Prevention of Strongylidae infections in the horse]. / Preventie van Strongylidae infecties bij het paard.

Proceeding on the epidemiology of Strongylidae infection in horses, the effects of combined anthelmintic treatment and moving to non-contaminated pasture in July were studied during 1981 and 1982. This strategy for control was found to be effective as was shown by faecal examination, pasture larval counts and post-mortem worm counts. No significant contamination or worm infection developed in the group of treated ponies moved to celan pasture after July 1. On the permanently grazed pasture the same pattern of infection developed as on the pasture where up to July 1 ponies and subsequently sheep had grazed.

[Effect of albendazole and cambendazole on the egg output of Strongylidae in horses]. / De invloed van albendazole en cambendazole op de ei-uitscheiding van Strongylidae bij het paard.

The anthelmintic activity of albendazole (5 mg/kg) and cambendazole (20 mg/kg) was compared in twenty-two naturally infected horses and ponies, which were divided into two equal groups. Comparison was based on the faecal strongyle egg counts. On the fourteenth and twenty-eighth days after the first treatment in May, the reductions in faecal egg counts were 99.1 and 93.9 per cent respectively in the albendazole group and 88.0 and 75.7 per cent respectively in the cambendazole group. Within fourty-two days after treatment had been initiated, the mean EPG (eggs per gram) had been restored to the initial level in both groups. The same animals were treated again in August; this time, however, the anthelmintics were changed. In the albendazole group, the faecal egg counts were reduced by more than 95 per cent and, in the cambendazole group, by not more than 84.1 per cent. As the horses were not weighed, the disappointing results obtained with camendazole may have been due to low a dosage, although resistance cannot be ruled out. When the trial was concluded in October, within fifty-six days after treatment in August, the faecal egg counts continued to be remarkably low in both groups: 8.3 per cent of the average EPG prior to treatment in the albendazole group and 21.2 per cent in the cambendazole group. As the trials did not provide any evidence to suggest a larvicidal action of the two anthelmintics, the difference with the results of treatment in May is attributable to an inhibitory effect on the development of the infective Strongyle larvae, specifically Cyathostominae, ingested during the summer.

The efficacy of ivermectin against Strongyloides westeri in foals.

Seven foals naturally infected with Strongyloides westeri were injected intramuscularly with ivermectin at a dosage rate of 200 mcg per kg body weight. No adverse effects to treatment were observed. Weekly faecal egg counts showed a greater than 99 per cent reduction of S. westeri egg output compared with 7 untreated foals during the 21 days following treatment.