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A technique for the cryopreservation of third-stage larvae of Strongylus vulgaris is described. Infective larvae of S. vulgaris were exsheathed in a 0.16% sodium hypochlorite solution and then transferred into cryotubes containing 0.09% saline. The samples were stored in the gas phase of liquid nitrogen.
Proliferation in vitro of peripheral blood lymphocytes both from horses infected with Strongylus vulgaris and from helminth-free ponies was observed in the presence of extracts of the fourth and fifth stage larvae and adults of S. vulgaris. In addition, S. vulgaris extracts induced transformation in cultures of peripheral blood lymphocytes from sheep and dogs and in mouse spleen cell cultures. Nylon wool non-adherent, T cell enriched fractions of lymphocytes from both mice and horses were stimulated by the S. vulgaris larval mitogen while no proliferation was observed in cultures containing nylon wool adherent, B cell enriched fractions. Macrophage co-operation appeared not to be necessary for S. vulgaris mitogen-induced transformation of spleen cells. The S. vulgaris mitogen stimulated a subpopulation of mouse spleen cells different from those responsive to PHA, Con A and LPS. These cells might be T helper cells since B cells were stimulated to proliferate in the presence of both T cells and S. vulgaris larval mitogen. In addition, the supernatant of in vitro cultured larvae of S. vulgaris induced slight, but significant transformation of equine peripheral blood lymphocytes. Therefore, it is possible that the S. vulgaris mitogen released by both viable parasites and degenerating larvae might induce T cell dependent production of immunoglobulin in vivo and account for the beta-globulinaemia, of which IgG(T) is a major component, in S vulgaris infected horses.
Between August 1978 and July 1979 the anterior mesenteric artery and its branches were collected regularly from adult horses and examined for Strongylus vulgaris larvae. The incidence of infection varied from 55 to 100% (annual mean 80%). The mean monthly number of larvae ranged form 3 to 22 with an annual overall mean of 13. The arterial infection was at its minimum in December to January, rose gradually to attain the peak in June and declined thereafter. These observations indicated that S. vulgaris is an annual species in Morocco, infection occurring during the rainy season (November-April), the heavy arterial population in spring and adult population during autumn and winter.
An indirect fluorescent antibody assay (IFA) was developed using Strongylus vulgaris third stage larvae (L3) as antigens. Observations using the IFA indicate that a species-specific antibody response to S. vulgaris L3 develops in S. vulgaris-infected ponies and that some surface L3 antigens are shared by adult worms. Sequential antibody levels against S. vulgaris were measured in strongyle-naive and in immune ponies following initial and challenge infections using the IFA and an indirect hemagglutination assay (IHA). Antibody levels measured by IFA increased faster following initial infections than did levels measured by IHA. Antibody levels appear to increase following challenge infections of immune ponies when measured with the IFA, but not with the IHA. Significant differences in antibody titers were not seen between ponies which developed colic following challenge infections and those that did not develop colic. Antibodies were not detectable in ponies unexposed to larval migrations, but which received surgical implantation of S. vulgaris adults into the cecum.
The four species of the genus Strongylus Müller, 1780, namely, S. equinus, S. edentatus, S. vulgaris and S. asini were examined with the scanning electron microscope (SEM). The buccal cavity of each species was cut transversely and longitudinally and the scanning electron micrographs illustrate the shape of the buccal cavity and the dorsal gutter. The micrographs also show the presence of denticles near the oral rim of the buccal cavity of S. edentatus and the shape of the teeth in the buccal cavity of S. equinus, S. vulgaris and S. asini. The bursa of the four species has three symmetrical lobes, one dorsal and two lateral. The length of the three lobes in relation to each other varies between the species. The genital cone has a single large ventral papilla, a pair of dorsal raylets, a median dorsal cuticular appendage and a variety of cuticular appendages lateral and ventral to the cloaca. The development and arrangement of these components differs between the four species of the genus and can be used as a supportive character for specific identification.
A study of Ctenocephalides felis strongylus and of its effect on the performance of sheep and goats was made. Fleas showed more affinity for sheep than goats. There was a high concentration between lamb and kid mortality and the degree of flea infestation. The clinical effects of flea infestation are highlighted. The factors responsible for flea infestation and its control are discussed.
Infestation of exotic and indigenous breeds of cattle, sheep and goats with Ctenocephalides felis strongylus in a dairy farm is reported. Many of the fleas were engorged with fresh blood. Traps set up in the pens caught two bush rats which were heavily infested with the same flea species, most of which were also engorged with fresh blood. It was suggested that the fleas might have been introduced into the pens by rodents such as bush rats.
An attempt was made to control or eliminate Strongylus vulgaris from a closed group of three horses at pasture near Perth, Western Australia, by dosing with ivermectin on four occasions during the time of year when it was believed that environmental conditions would eliminate all the non-parasitic stages of that species. At necropsy, five months after the last dose of anthelmintic and after continually grazing the same pastures, no S vulgaris or arterial lesions were found in those horses and S edentatus, Draschia megastoma and Habronema species were also almost completely eliminated.
The concentrations of haptoglobin, immunoglobin (Ig)G(T) and IgG were measured in the serum of four previously parasite-free pony yearlings following a single dose of 700 (Group H) or 200 (Group L) stage three Strongylus vulgaris larvae (L3) and following a reinfection with the same doses 34 weeks later. The results are compared with an uninfected control pony. The haptoglobin concentration increased during Weeks 1 to 6 and 14 to 17 after infection in the serum of the ponies receiving 200 L3, but in only one pony dosed with 700 L3 (during Weeks 1 to 16). The serum haptoglobin also increased during the first seven weeks after the second infection, in three of the four ponies following the second dose of larvae. The serum IgG(T) concentration started to increase from Week 6 or 9 in the ponies given 700 L3, reaching peaks of 44 and 32 g/litre respectively, eight to nine weeks later, compared with a peak of 16 g/litre 20 to 22 weeks after infection in ponies dosed with 200 L3. The IgG(T) concentration increased to a maximum of 25 g/litre in the serum of only one of the four ponies after the reinfection. The serum IgG concentration in all ponies increased nearly twofold during the first eight weeks after both the primary and secondary dose of larvae. It is concluded that the measurement of specific proteins is more reliable and quicker than the electrophoretic separation and quantitation of protein bands, in tracing changes in serum proteins following the artificial infection of ponies with S vulgaris larvae.
The concentrations of serum proteins (beta 1, beta 2, gamma, alpha 1, alpha 2 globulins and albumin) and absolute numbers of eosinophils, neutrophils and lymphocytes were examined in 64 naturally infected horses and ponies in which the number of larvae of Strongylus vulgaris in the cranial mesenteric artery and the severity of the lesion of verminous arteritis could be determined. The horses were grouped according to the number of larvae found and the severity of the arteritis. The results demonstrated that, although some significant deviation from a random distribution occurred in certain of the values (chi 2 test), there was considerable individual variation in the values obtained for individual animals within groups and overlap of the range of values between groups. Also the number of larvae present in the artery did not necessarily accurately reflect the severity of the arterial lesion. Thus, the parameters examined could not be used reliably to estimate the intensity of infection with S vulgaris in an individual animal.
Eighteen parasite-free pony foals were infected orally with 500 third stage larvae of Strongylus vulgaris. At 56 days after infection, six ponies were treated with intramuscular ivermectin (22, 23-dihydroavermectin B1); six were treated with oral ivermectin; and six were not treated. Necropsy was done 91 days after infection to study the pathologic effects of migrating S. vulgaris larvae and to determine the efficacy of ivermectin in attenuation of S. vulgaris-induced lesions. Larval migration induced eosinophilic inflammation of the liver, spleen, mesenteric, colic and cecal lymph nodes, and small and large intestine. Previously unreported parasitic lesions included eosinophilic pneumonia with eosinophilic granulomas and pulmonary lymphoid nodules. S. vulgaris larvae were observed in eosinophilic granulomas in the lung, epicardium, liver, and intestinal serosa. Injectable and oral ivermectin formulations were equally effective in reduction of these lesions.
The clinical signs, pathology and clinical pathology associated with single experimental infections of Strongylus vulgaris in worm-free pony foals are described. The major clinical signs which became apparent in the infected foals during the first three weeks were pyrexia, anorexia, dullness and abdominal pain. Within the first two weeks of infection lesions were confined to the intestine and terminal branches of the intestinal arteries and consisted of mucosal, submucosal and serosal haemorrhage together with arteritis of submucosal and serosal arteries and also a marked inflammatory reaction. The main lesion seen three weeks after infection was gross thrombosis of the anterior mesenteric artery or one of its major branches. On section these affected arteries showed marked intimal thickening with infiltration of plasma cells, lymphocytes, macrophages and neutrophils. Between one and four months after infection the gross lesions were predominantly in the arteries and consisted of fibrous thickening of the arterial wall and thrombosis associated with the presence of developing fourth stage larvae. Four months after infection the arterial lesions were still prominent and microscopically there was fibrosis of the wall of the affected artery with wide-spread disruption of the intima. In the adventitia organised thrombi were apparent in the vasa vasorum and resulted in the obliteration of their lumina. The typical lesion associated with the return of fifth stage larvae to the intestine was nodule formation in close proximity to thrombosed terminal intestinal arteries and sections of parasites were seen in the intestinal wall surrounded by neutrophils and necrotic debris. By nine months after infection the arterial lesion had healed, but histologically there was fibrosis of the intima and macrophages containing haemosiderin were seen in the arterial wall. The most significant haematological findings during the experimental period were a marked polymorphonuclear leucocytosis and an increase in the number of circulating eosinophils in the infected animals. Also marked was an increase in the serum globulin levels of the infected foals.
Four pony foals were inoculated with Strongylus edentatus infective larvae and on days 3 and 4 postinfection two of the ponies were treated with thiabendazole, each at the rate of 440 mg/kg of body weight. Total circulating eosinophil counts in untreated ponies increased to over 1700 per cu mm after the second week postinfection. In the treated ponies as well as in an uninfected untreated pony eosinophil counts did not increase beyond 100 per cu mm. At necropsy on day 35 postinfection the cecum, colon and omentum of treated ponies were normal and few tracks were present on the surface of the liver. In untreated ponies nodules were observed on the serosal surface of the cecum and right ventral colon and white foci and tracks were numerous on the surface of the liver. A total of 53 fourth stage larvac was recovered from the livers of the thiabendazole treated ponies and 1194 from the untreated ones.
Twenty pony foals (reared worm free), 6.5 to 10 weeks of age, were inoculated with Strongylus vulgaris and allocated to 5 groups, each with 4 foals. One week after inoculation, 1 group of 4 foals was given oxfendazole (OFZ) at a dosage rate of 10 mg/kg of body weight, another group was given 2 such treatments 48 hours apart, and a 3rd group was given a placebo. All treatments were administered by stomach tube. Three weeks later, foals were euthanatized and necropsied in a test for efficacy against early 4th-stage larvae. Oxfendazole was 80% and 94.9% effective against early 4th-stage S vulgaris with 1 and 2 doses, respectively. A 4th group of 4 foals was given 2 treatments of OFZ, 48 hours apart, about 8 weeks after inoculation, and a 5th group was given a placebo. These foals were euthanatized and necropsied 5 weeks after treatment in a test for efficacy against later 4th-stage larvae. Two doses of OFZ were 96.6% effective against later 4th-stage larvae.
Five weanling ponies were subjected to an intensive 6-week deworming program after which 4 Ag-AgCl bipolar electrodes were implanted surgically on the distal ileum. For 3 hours each day for 5 consecutive days, ileal myoelectrical activity was recorded from fed ponies under 3 sequential conditions: preinoculation, after oral administration of 1,000 killed Strongylus vulgaris infective larvae (3 ponies), and after oral administration of 1,000 live S vulgaris infective larvae. Recordings were analyzed for slow wave frequency, percentage duration of phases I, II, and III of the migrating myoelectrical complex (MMC), and the frequency of distinct, rapidly migrating action-potential complexes within phase 2 of the MMC. After administration of live and killed infected 3rd-stage larvae, there was a marked increase in the number of disrupted phase III complexes, and a significant (P less than 0.001) increase in the number of migrating action-potential complexes. In addition, after inoculation of live 3rd-stage larvae, there was a significant increase (P less than 0.001) in the percentage of time that the MMC was occupied by prolonged periods devoid of spike activity (phase I). The results indicate that S vulgaris larval mucosal penetration and submucosal migration can cause changes in ileal myoelectrical activity that could cause colic, and that larval antigen alone within the lumen may disrupt ileal motility.
Two controlled tests were conducted in equine foals and yearlings to determine the optimal oral dosage and the duration of activity of closantel for the prevention of Gasterophilus spp larval infections. Additional data were collected on the activity of closantel against Strongylus vulgaris larval infections. In experiment 1, 12 foals and 12 yearlings were equally allocated to 4 experimental groups, and were given oral treatments with closantel at dosages of 0 (nontreated controls), 2, 5, or 8 mg/kg of body weight every 2 months during bot season. The foals and yearlings were allowed to graze on open pasture throughout the experiment to provide a natural source for bot and helminth infections. All animals were euthanatized and necropsied 6 weeks after the final treatment. Closantel was highly effective (98.6% to 100%) at all doses in preventing Gasterophilus spp larval infections in the foals, but only the 8 mg/kg dose had significant (P less than 0.05) activity (99.7%) in the yearlings. This dose also significantly reduced the numbers of 4th-stage and immature adult S vulgaris (86.0%) in the mesenteric arteries as compared with nontreated controls. In experiment 2, 9 foals and 9 yearlings received a single oral treatment of 8 mg of closantel/kg of body weight; 3 foals and 3 yearlings were kept as nontreated controls. Groups of 6 treated (3 foals, 3 yearlings) and 2 control (1 foal, 1 yearling) animals were euthanatized and necropsied 1, 2, and 3 months after treatment. Closantel remained effective for 2 months in preventing infections of G intestinalis larvae in these foals and yearlings. Clinical signs of toxicosis were not observed in the treated animals of either study.
Pony foals inoculated with infective Strongylus edentatus larvae were monitored for clinical signs and selected blood changes and were examined at necropsy from two to 56 days postinfection. Larvae penetrated the intestine and reached the liver intravenously before 40 hours postinfection. Occasional thrombi and larval tracks associated with the intima of cecal and colic veins suggested aberrant paths. Larvae in the liver doubled in width between seven and 15 days postinfection and a sudden increment in circulating eosinophils occurred between 11 and 15 days. These changes were probably associated with the third molt. At 30 days fourth stage larvae were migrating in the liver; at 42 days they were present in the hepatorenal ligament. White foci were observed in the liver from two to 56 days. They contained mononuclear cells and eosinophils and later necrotic cores of eosinophils. By one month foci were overshadowed by tortuous tracks of migrating larvae. Aberrant larvae in the lungs were confined in granulomas. Massive granulomas in the wall of the cecum and colon contained small larvae which were probably inhibited by antibody associated with the third molt. Severe disruption of omental architecture and adhesions involving the intestine occurred several weeks after infection.
Twelve pony foals (reared worm-free) were inoculated with Strongylus vulgaris. Approximately 8 weeks later, 4 of the foals were given fenbendazole (10% suspension) at a dosage rate of 10 mg/kg of body weight daily for 5 days and 4 foals were given the suspension at a dosage rate of 50 mg/kg daily for 3 days; the remaining foals were given a placebo. All treatments were administered by stomach tube. Fenbendazole was 99.6 and 97.9% effective in the 2 treatment groups, respectively, in eliminating later 4th-stage S vulgaris larvae located near the origin of major intestinal arteries. On microscopic examination of the ileocolic artery from fenbendazole-treated foals, a few larval remnants were found beneath the tunica intima in small organized mural thrombi overgrown with endothelium. It would appear that larvae are rapidly destroyed after administration of fenbendazole. A pony foal reared on pasture and with arteriographic evidence of arteritis of the cranial mesenteric and ileocolic arteries was treated with fenbendazole (10% suspension) by stomach tube at a dosage rate of 50 mg/kg of body weight daily for 3 days. By arteriographic examination made 4 weeks later, there was evidence of regression of the lesion, and at necropsy done a week later, there was no arteritis or larvae in the lumen of those arteries.