Hematological and coagulation profiles in dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866).

Hematological and coagulation profiles were studied in crossbred dogs experimentally infected with Angiostrongylus vasorum. Two groups of five dogs were experimentally inoculated with 50 and 100 third stage infective larvae (L(3)) of A. vasorum per kilogram of body weight. A third group of five uninfected animals was used as control. One sample of 10 ml of blood was collected from each animal on the 10, 20, 30, and 45 days after inoculation (dai) and at 30-day intervals thereafter for the remainder of the 210-day experimental period. The blood sample was used for the complete hemogram and platelet count, as well as measurements of prothrombin time, partial thromboplastin time and factors V and VIII. Anemia was observed in infected dogs, 6 weeks after the infection. The eosinophils presented peaks in four periods after infection. Thrombocytopenia became accentuated on the 72 dai. Decreased prothrombin time activity and increased partial thromboplastin time were observed at the 6 and 9 weeks after infection and decreased of factors VIII and V activities occurred from 4 to 6 weeks after infection. It may be conclude that infection by A. vasorum in dogs may cause a discrete anemia during the acute phase which is probably regenerative. In addition, important hemostatic alterations due to the infection suggest a chronic intravascular consumption coagulopathy.     

Coagulation profiles in dogs with congenital portosystemic shunts before and after surgical attenuation

BACKGROUND: Serious postoperative hemorrhage has been reported in dogs after closure of congenital portosystemic shunts (CPS). HYPOTHESIS: In dogs with portosystemic shunting, low coagulation factor activity is responsible for coagulopathy, which can cause complications after surgery. ANIMALS: Thirty-four dogs with CPS and 39 healthy dogs. METHODS: In a prospective study, coagulation times, platelet count, and the activity of 8 coagulation factors were measured in dogs before and after surgical shunt attenuation and in 31 healthy dogs. The effect of abdominal surgery on hemostasis was determined at ovariectomy in 8 healthy dogs. RESULTS: Dogs with CPS had lower platelet counts, lower activity of factors II, V, VII, and X, and increased factor VIII and activated partial thromboplastin time (APTT) compared to healthy dogs. After surgical attenuation, dogs with CPS had decreased platelet counts and activity of factors I, II, V, VII, IX, X, and XI and a prolonged prothrombin time (PT). Ovariectomy resulted in decreased activity of factors VII and X. Six weeks after surgery, portosystemic shunting persisted in 9 of 30 dogs, with no improvement of hemostatic values. CPS dogs without shunting had improved coagulation times and increased activity of factors II, V, VII, and X. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with CPS have lower activity of clotting factors compared to healthy dogs, resulting in a prolonged APTT. Surgical attenuation of the shunt results in increased abnormalities in coagulation times and factors immediately after surgery. Hemostasis is normalized after complete recovery of shunting after attenuation, in contrast to dogs with persistent shunting.     

Clinical and hematologic effects of experimental infection of dogs with recently identified Babesia gibsoni-like isolates from Oklahoma

OBJECTIVE: To characterize clinical and hematologic responses in dogs following experimental inoculation with Babesia gibsoni-like isolates from infected dogs in Oklahoma. DESIGN: Prospective study. ANIMALS: 6 mixed-breed dogs. PROCEDURE: 2 dogs were inoculated with organisms from a naturally infected dog, and 3 were inoculated with organisms from a second naturally infected dog (1 of these 3 dogs was splenectomized 1 week prior to inoculation). One dog was not inoculated. Complete blood counts were performed weekly. RESULTS: In the 5 dogs inoculated with organisms, parasites were initially detected 1 to 5 weeks after inoculation, and severity of parasitemia peaked with 1.9 to 6.0% of RBC infected by 4 to 6 weeks after inoculation. Parasitemia was easily detectable (> 0.1% of RBC infected) for 3 to 4 weeks. Clinical abnormalities included lethargy, fever, and pale mucous membranes but were mild to nearly inapparent in 2 dogs. All dogs developed regenerative anemia and marked thrombocytopenia. Thrombocytopenia developed before and lasted longer than the parasitemia. Profound but transient neutropenia was detected in some dogs. The splenectomized dog developed more severe parasitemia and anemia and more pronounced clinical abnormalities. Three dogs with intact spleens recovered without treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that 2 or more genotypically distinct, but morphologically identical, small Babesia parasites can infect dogs in the United States. Compared with infection with small Babesia parasites from California, infection with these isolates resulted in less severe parasitemia and clinical abnormalities. Parasitemia was transient, indicating that identification of organisms in blood smears may be difficult in some dogs.     

Patent Toxocara canis infections in previously exposed and in helminth-free dogs after infection with low numbers of embryonated eggs

The outcome of Toxocara canis infections in the canine host depends on the migratory pathway of parasite larvae (somatic or tracheal) which is considered to be related to the host's age and its immune status. However, field studies attest high prevalences of patent T. canis infections in adult animals. The controlled induction of patent infections with low doses of embryonated eggs was investigated in 18 beagles in a 7-month study until their 16th life month. The animals were assigned to three groups, each consisting of three vertically infected dogs (with a short patent infection as pups before anthelmintic treatment) and three helminth-free dogs. At study days 10 and 40, the animals of groups 1 and 3 were given each 100 embryonated T. canis eggs. In each case, group 1 was treated 10 days post-infection with Milbemax, while dogs of group 3 remained untreated. Control group 2 was not experimentally infected but treated as group 1. Two weeks after first egg administration, a sharp increase of specific antibody reactions in ELISA and increased eosinophilic counts indicated larval invasion in all infected dogs. 42-56 days following first infection, patent infections were detected coproscopically in all animals of group 3, but in none of the uninfected dogs (group 2) or the infected and treated dogs (group 1). Following a 3-month observation period, all animals of the three groups were treated with piperazine citrate to eliminate intestinal infections and all were administered 100 embryonated eggs. Subsequently, patent infections developed in animals of all groups: in one of the infected and treated animals of group 1, in five of the so far not infected control group 2 and in four of the dogs with previous patent infections (group 3). Susceptibility to patent infections was not significantly altered in T. canis-free dogs compared to dogs with previous patent infection (vertically acquired or experimentally induced). However, dogs of group 1 treated with Milbemax after repeated egg administration developed a significantly increased resistance to patent infections as compared to control dogs (group 2). Observed prepatency periods were between 40 and 56 days and did not differ in the three groups. Even in urban areas, facing high infection pressure with Toxocara eggs maintained by a high dog and fox population, dogs of all ages are at risk to develop patent T. canis infections.     

Ivermectin treatment of naturally acquired and experimentally induced Strongyloides stercoralis infections in dogs.

Treatment of Strongyloides stercoralis infection was investigated in 2 dogs with naturally acquired, chronic-active infections, and in 3 dogs with corticosteroid-enhanced, experimentally induced hyperinfections. A single oral dose of ivermectin was given to naturally infected (200 micrograms/kg of body weight) and experimentally infected (800 micrograms/kg) dogs. Five dogs with experimental hyperinfections served as controls. Dogs with naturally acquired infections ceased to shed first-stage larvae in the feces 1 week after treatment, but 1 dog had recrudescence and required a second dose. Ivermectin was 100% effective in removing adult S stercoralis from the intestinal tract of the experimentally infected dogs, but it was not effective in removing third-stage larvae from parenteral sites. Ivermectin-treated dogs had few intestinal parasites of any stage, whereas at necropsy, 4 of 5 experimentally infected dogs not treated had massive infections (greater than 100,000 adults, greater than 92,000 larvae) in the intestinal tract, and 3 of 5 had larvae (greater than 2,500) in parenteral sites.     

The immune response of pigs to infection with the stomach worm Hyostrongylus rubidus (Hassall and Stiles, 1892): I. Antibody production in response to primary infection

Pigs infected with single doses of third stage Hyostrongylus rubidus larvae showed a detectable circulating agglutinin response as early as 2 days post infection. Although titres fluctuated considerably before patency, a trend in antibody production could be detected which followed the active development and growth of larvae in the gastric mucosa. At patency, titres showed a large, rapid increase followed by an equally dramatic decrease, after which titres remained stable or increased gradually.Pigs infected with adult worms did not show a detectable agglutinin response until 7 days post infection or later. Little fluctuation in titre was observed.Response variation between different levels of infection was great as was the variation between different litters subjected to the same infection regime. No statistical difference could be found in total antibody production between pigs infected with different larval numbers at any time during the duration of the trial.     

Immunity to parasitic bronchitis of yearling cattle treated with ivermectin during their first grazing season

A group of 12 winter-born calves was divided into two groups of six. During the following summer one group grazed on pasture infected with Dictyocaulus viviparus, and was treated with ivermectin injections at three, eight and 13 weeks after turn out. The other group remained housed. Both groups were housed during the winter and then together with a group of younger calves were challenged with a trickle infection of D viviparus larvae at the rate of 25 third stage larvae/kg bodyweight for one month and then slaughtered. The group which had been exposed to previous infection was least affected by parasitic bronchitis and on the basis of serological titres and worm burdens had developed resistance to the challenge infection. The other older group was also more resistant than the younger calves.     

Comparison of a levamisole sustained-release bolus and ivermectin treatment to prevent bovine lungworm infection

The efficacy of a levamisole sustained-release bolus to prevent parasitic bronchitis in calves in their first grazing season was compared to ivermectin treatment at three, eight and thirteen weeks after turn out. Contamination of the pasture was established by experimentally infected seeder calves. Twenty calves were split into two groups. Ten calves of one group received a bolus at the start of the experiment. In the other group the calves were treated with ivermectin at 21, 56 and 91 days. Two principal calves from each group were killed during the experiment to study histopathological changes. Pairs of tracer calves were introduced on both pastures at intervals of four weeks throughout the grazing period. The permanent calves were challenged with lungworm larvae at housing and slaughtered four weeks later. Both systems prevented parasitic bronchitis. Larval output was completely reduced in the ivermectin-treated calves while all bolus-treated calves excreted larvae at certain times. The highest group average was 4 larvae per gram faeces. Eosinophilia, ELISA-titres and histopathological changes confirmed the differences in larval uptake. Challenge infection was not successful in either group and no worms were found at slaughter. Weight gain was significantly different at housing in favour of the ivermectin-treated calves, but after challenge this was reduced due to a higher weight gain in the bolus-treated calves. The practical consequences of the results have been discussed.     

Specific antibody responses to Taenia hydatigena, Taenia pisiformis and Echinococcus granulosus infection in dogs

Groups of dogs reared free of both nematodes and cestodes were infected with Taenia hydatigena, Taenia pisiformis or Echinococcus granulosus. After infections with the Taenia spp became patent, dogs were purged to remove the worms. They were later reinfected and the second infections again removed by purging after patency. A group of 3 uninfected worm free dogs was kept as age-matched controls. The dogs were bled at intervals of 5 days and their serums tested for antibodies using the enzyme-linked immunosorbent assay (ELISA) with excretory/secretory (ES) antigens collected during in vitro incubation of evaginated scoleces (scolex ES antigen) and oncosphere antigens. Antibodies to scolex ES antigen were detected by 3 weeks after infection with each cestode species whereas antibodies to oncosphere antigen were not detected until about one week after eggs were found in the faeces of the infected dogs. Antibody responses to both oncosphere and scolex ES antigens decreased rapidly following removal of the worms by purging. Uninfected control dogs were invariably negative to both oncospheral and scolex ES antigens. There were cross-reactions between the serums from dogs infected with T. pisiformis and T. hydatigena when tested with scolex ES antigens, but oncospheral antigens showed a high degree of species specificity. Scolex ES antigens from E. granulosus were compared with those prepared from T. hydatigena and T. pisiformis for their ability to discriminate between antibodies in serums collected from dogs 31 and 32 days after infection with 100,000 protoscoleces of E. granulosus or dogs infected with Taenia spp.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of fenbendazole on Toxocara canis larvae in tissues of infected dogs.

The effect of fenbendazole therapy was studied in six dogs fed 10,000 embryonated Toxocara canis eggs. At 47 days after they were fed T canis, four dogs were given fenbendazole in two divided doses totaling 50 mg/kg of body weight each day for 14 days. Two infected dogs were not given fenbendazole. All dogs were necropsied at the end of treatment and the foci were counted in the lungs; their skeletal muscles were digested in 1% trypsin for the recovery of larvae. The T canis larvae were not recovered from the skeletal muscle of the four infected dogs treated with fenbendazole; 15 and 42 larvae/100 g of skeletal muscle were recovered from the two nonmedicated infeected dogs. The number of grossly visible foci on surfaces of lungs in treated dogs was markedly less than in the nonmedicated infected dogs. The results indicate that fenbendazole might be effective in preventing prenatal infection in dogs.     

A blind test of the serological response of dogs to infection with Taenia ovis

Fifty six dogs of mixed age and sex were acquired from farms in the Otago/Southland region, and maintained at the Hydatid Research Unit, Taieri, where 43 were each fed two Taenia ovis cysts. All were bled fortnightly for six or 12 weeks. Coded sera were sent to Wallaceville Animal Research Centre for testing using ELISA, with antigen from T. ovis scoleces. Dog treatments were identified after all tests were complete. A discriminant level was derived from the mean absorbance value plus three standard deviations of 56 sera taken at time zero and 78 sera from serially bled uninfected dogs. None of these 134 sera registered as a false positive using this discriminant level. The data showed no significant deviation from normality, and the expected frequency of the occurrence of false positives is therefore less than 0.14%. Four weeks after infection 63% of dogs proved to be infected were serologically positive, rising to 78% after 6 weeks. When worms were removed by anthelmintic treatment, ELISA absorbance levels decreased. Four weeks after removal 70% of previously infected dogs remained positive, decreasing to 30% after 6 weeks.

Six weeks after infection the sensitivity of the test was 78%, and the specificity 63%. However, if dogs with positive ELISA absorbance levels, but which did not purge worms, were regarded as having had worms, the respective figures would be 82% and 100%. The latter figures are similar to our previously published laboratory results. The test is of comparable efficiency to arecoline purgation for surveillance, and has the additional advantage of detecting infection in the majority of those dogs that have been infected for three weeks or more but fail to pass worms on purgation, and a substantial proportion of those infected dogs that were treated by their owners prior to presenting them for purgation in order to avoid detection of infection.     

A blind test of the serological response of dogs to infection with Taenia ovis

Fifty six dogs of mixed age and sex were acquired from farms in the Otago/Southland region, and maintained at the Hydatid Research Unit, Taieri, where 43 were each fed two Tueniu ovis cysts. All were bled fortnightly for six or 12 weeks. Coded sera were sent to Wallaceville Animal Research Centre for testing using ELISA, with antigen from T. ovis scoleces. Dog treatments were identified after all tests were complete. A discriminant level was derived from the mean absorbance value plus three standard deviations of 56 sera taken at time zero and 78 sera from serially bled uninfected dogs. None of these 134 sera registered as a false positive using this discriminant level. The data showed no significant deviation from normality, and the expected frequency of the occurrence of false positives is therefore less than 0.14%. Four weeks after infection 63% of dogs proved to be infected were serologically positive, rising to 78% after 6 weeks. When worms were removed by anthelmintic treatment, ELISA absorbance levels decreased. Four weeks after removal 70% of previously infected dogs remained positive, decreasing to 30% after 6 weeks. Six weeks after infection the sensitivity of the test was 78%, and the specificity 63%. However, if dogs with positive ELISA absorbance levels, but which did not purge worms, were regarded as having had worms, the respective figures would be 82% and 100%. The latter figures are similar to our previously published laboratory results. The test is of comparable efficiency to arecoline purgation for surveillance, and has the additional advantage of detecting infection in the majority of those dogs that have been infected for three weeks or more but fail to pass worms on purgation, and a substantial proportion of those infected dogs that were treated by their owners prior to presenting them for purgation in order to avoid detection of infection.     

Response of sheep to challenge infection with Fasciola hepatica

Sheep were infected with 100 metacercariae of Fasciola hepatica and reinfected 16 weeks later with a further 100 metacercariae. Serum samples were taken weekly for 36 weeks after primary infection. Serum was assayed for the presence of the enzymes glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (gamma-GT), as indicators of liver and bile duct damage respectively, and for levels of precipitating antibody. Antibody and GLDH levels rose following the primary infection but fell after patency had been reached . A peak in gamma-GT activity was associated with the onset of patency. After the challenge infection levels of both enzymes rose substantially and there were persistent fluctuations in activity. Antibody levels did not rise markedly following challenge but fluctuated at low levels until autopsy, 20 weeks after challenge. There was no resistance to challenge judged by worm recoveries at autopsy. It is suggested that the presence of adult flukes in the bile ducts suppresses the antibody response to challenge infection. Tissue damage, which is shown by fluctuations in GLDH and gamma-GT levels after adult flukes have become established in the bile ducts, is considered to be due to the feeding activity of adult flukes and the deposition of immune complexes in the liver parenchyma.     

The response of Suffolk lambs to an escalating experimental infection with Dictyocaulus filaria.

Infection of Suffolk lambs with Dictyocaulus filaria larvae beginning at two months of age with one larva per day, six days per week, and doubling every four weeks to reach a final infection rate of 64 larvae per day gave rise to a pattern of larval excretion in the faeces which approximated to that seen in naturally infected Suffolk lambs in Midlothian in 1974. Several of the lambs were still infected with adult D filaria and were excreting larvae in their faeces 28 weeks after the infection began. However, when some of the lambs were challenged at this time, those which had received the previous escalating infection were resistant to the challenge infection.     

Measurement in vitro of a larval migration inhibitory factor in gastrointestinal mucus of sheep made resistant to the roundworm Trichostrongylus colubriformis

Lambs were challenged by dosing with 2500 T. colubriformis larvae per day for 34 weeks, rested for 4 weeks and then re-challenged with the infective larvae for a further 10 weeks. A technique for the measurement of inhibition of larval migration from agar gels was used to investigate the antiparasitic activity of mucus, obtained from the small intestine and abomasum of the lambs, at the end of the re-challenge period. Measurements were also made on ileal digesta samples collected at different times during the development of the initial resistance, the rest period and the re-challenge period, and on faeces collected during the re-challenge infection. The mucus from the small intestine and abomasum paralysed and inhibited larval migration from agar gels significantly more (P less than 0.01 and P less than 0.05, respectively) than corresponding mucus from parasite-free control animals. This inhibitory activity was also detected (P less than 0.01) in the ileal digesta of infected animals from Week 6 of primary dosing. The magnitude of the inhibition in the ileal digesta increased with time during the infection period and was again detected during re-infection (P less than 0.01). It was not detectable in resistant sheep towards the end of the rest period. Slight inhibitory activity was detected in faeces after 2 weeks of re-infection. These observations suggest that substances secreted into the lumen of the small intestine of infected animals are responsible for resistance against ingested larvae.     

Pulmonary Artery Thrombosis in Experimental Angiostrongylus vasorum Infection Does Not Result in Pulmonary Hypertension and Echocardiographic Right Ventricular Changes

Background: Dogs experimentally inoculated with Angiostrongylus vasorum develop severe pulmonary parenchymal lesions and arterial thrombosis at the time of patency. Hypothesis: A. vasorum‐induced thrombosis results in arterial hypoxemia, pulmonary hypertension (PH), and altered cardiac morphology and function. Animals: Six healthy Beagles experimentally inoculated with A. vasorum. Methods: Thoracic radiographs and arterial blood gas analyses were performed 8 and 13 weeks postinoculation (wpi) and 9 weeks posttherapy (wpt). Echocardiography was done before and 2, 5, 8, 13 wpi and 9 wpt. Invasive pulmonary artery pressure (PAP) measurements were obtained 8 wpi. Two untreated dogs were necropsied 13 wpi and 4 treated dogs 9 wpt. Results: All dogs had patent infections at 7 wpi and clinical respiratory signs at 8 wpi. Moderate hypoxemia (median PaO₂ of 73 and 74 mmHg) present at 8 and 13 wpi had resolved by 9 wpt. Echocardiographically, no evidence of PH and no abnormalities in cardiac size and function were discernible at any time point. PAP invasively measured at 8 wpi was not different from that of control dogs. Severe radiographic pulmonary parenchymal and suspected thrombotic lesions at 13 wpi were corroborated by necropsy. Most histopathologic changes had resolved at 9 wpt, but focal inflammatory, thrombotic, and fibrotic changes still were present in all dogs. Conclusion: In experimentally infected Beagles, pulmonary and vascular changes induced by A. vasorum are reflected by marked radiographic changes and arterial hypoxemia. These did not result in PH and echocardiographic changes in cardiac size and function.     

Imidocarb: A chemoprophylactic experiment with Babesia canis

Summary

Eight dogs, given imidocarb dipropionate subcutaneously at a dose of 6 mg/kg, were challenged with a sporozoite stabilate of a French strain of Babesia canis, prepared from infected Dermacentor reticulatus ticks, 2, 3, 4 or 5 weeks after treatment. Three control dogs were similarly infected but not preventively treated. One of the controls and one of the dogs treated 5 weeks prior to challenge died of babesiosis. Prepatent and incubation periods were similar in treated and control dogs, and all dogs showed important reductions in the packed cell volume. Relaps'es were commonly seen after recovery from the initial reaction. Although further work is needed before a final conclusion can be drawn to whether imidocarb is suitable as a chemoprophylactic against B. canis infection, it can be used as a curative drug.     

Thrombocytopenia and platelet activity in dogs experimentally infected with Rangelia vitalii

The aim of this study was to evaluate the platelet count, coagulation time and platelet activity in dogs experimentally infected with Rangelia vitalii during the acute phase of the disease. For this study, 12 young dogs (females) were used, separated in two groups. Group A (uninfected control) was composed by healthy dogs (n=5), and group B consisted of R. vitalii-infected animals (n=7). After being inoculated with R. vitalii-infected blood, animals were monitored by blood smear examinations, which showed intra-erythrocytic forms of the parasite five days post-inoculation (PI). Blood samples were collected on days 0, 10, 20 and 30 PI. The material collected was placed in tubes containing EDTA for quantification of platelets, citrate anticoagulant platelet aggregation, and measuring the clotting time. Right after blood collection on days 10 and 20 PI, dogs were anesthetized for collecting bone marrow samples. A significant reduction (P<0.01) of the number of platelets was observed in R. vitalii-infected blood, when compared with uninfected dogs on days 10 and 20 PI. Additionally, macro-platelets were observed only in infected dogs. Prothrombin time and activated partial thromboplastin time did not differ between infected and uninfected dogs. The megakaryocyte count increased (P<0.01) significantly in infected dogs when compared with uninfected ones on days 10 and 20 PI. Platelet aggregation decreased (P<0.01) significantly in infected dogs in comparison to the control on days 10 and 20 PI. Therefore, rangeliosis in dogs causes a severe thrombocytopenia during the acute phase of infection. This platelets reduction probably occurred due to splenic sequestration and/or immune-mediated thrombocytopenia.