Age resistance and acquired immunity to Taenia pisiformis infection in dogs

An experiment was carried out using five groups of five beagle puppies each to measure age resistance to T. pisiformis infection and acquired immunity resulting from exposure to antigens released by adult tapeworms and/or oncospheres. Resistance was gauged by the number of worms established from challenge infections, and by the degree of development of those worms established (relaxed lenght, segment number, eggs per terminal proglottis).Age of puppies had a marked effect upon worm development, excluding the number of eggs per terminal proglottis, but not upon the number of worms established. Acquired immunity could not be demonstrated by any of the methods used.     

Prenatal and transmammary infections of Toxocara canis in dogs: effect of benzimidazole-carbamate anthelmintics on various developmental stages of the parasite

Fenbendazole and albendazole, given at a dose rate of 150 mg/kg for 3 days, produced a 90 per cent reduction in the numbers of second stage larvae of Toxocara canis present in the tissues of dogs although no reduction in the number of larvae found in the brains of infected dogs occurred with this treatment. The results suggest that a course of 3 day therapy with these anthelmintics should prevent prenatal infections in puppies. However, if infection is acquired by bitches during late pregnancy or early lactation, the transmammary route of infection becomes important. Therefore, anthelmintic treatment of the bitch prior to pregnancy will not prevent transmission of infection to her puppies should the bitch acquire a new infection of T. canis during pregnancy or early lactation. Alternatively, infection with T. canis can be controlled through the treatment of neonatal puppies for migrating larvae of T. canis. Treatment of newborn puppies with fenbendazole, albendazole or oxfendazole at a dose of 100 mg/kg for 2–3 days produced a 91–99 per cent reduction in the number of adult parasites found. In addition, a single dose of fenbendazole, given at a dose rate of 40 mg/kg, eliminated 93–96 per cent of adult T. canis from the intestines of 4–5-week-old puppies. These latter treatments would need to be repeated to eliminate completely the infection from puppies.     

Experimental Ascaris suum infection in the pig. Population kinetics following low and high levels of primary infection in piglets

Groups of piglets 20 to 28 days of age were dosed with 50 or 10 000 Ascaris suum eggs. The pigs were sacrificed from day 16 post infection onwards and worms present in the intestine were counted. Large numbers of immature parasites were recovered initially following the 10 000 egg dose, but the numbers decreased rapidly with the and worms were not found in pigs examined in the late prepatent phase. All pigletts infected with 50 eggs were found to harbour worms and the infection reached patency. The high level infections resulted in an eosinophil response which reached maximum values during the second week after infection. The low level infections which resulted in patency were not followed by any significant eosinophil response.     

Contamination of dog hair with eggs of Toxocara canis

Toxocara canis, the common intestinal nematode of dogs and foxes, is the parasite responsible for human toxocarosis. It has recently been shown that dogs may harbour eggs of the parasite in their fur. To further investigate this claim a population of 100 stray dogs was examined to establish the prevalence and intensity of adult toxocaral worm infection in the intestines and eggs harboured in the hair. A novel method of washing the eggs from the hair was used. Sixty-seven percent of dogs were found to have T. canis eggs on their hair with a mean egg retrieval of nearly 584 eggs per gram from positive dogs. The age of the dog was found to be the only significant factor to influence the prevalence and intensity of eggs, with 95% of all the eggs recovered found on puppies. Thirty-nine percent of dogs were found to have adult T. canis worms in their intestine, although a significantly higher percentage of puppies (80%) were infected with worms than adults (22.5%). Puppies also had more worms per infection than adults and have a strong positive correlation between egg and worms numbers whereas adults did not. These studies show that stray dogs, particularly puppies, potentially harbour considerable numbers of eggs on their hair, at densities far higher than those reported in the soil or the general environment.     

A Serological Survey of Dogs for Brucella canis in Southwestern Ontario

Sera from 2 000 dogs from southwestern Ontario were tested for antibodies to Brucella canis by a rapid slide agglutination test. The 100 positively reacting sera were tested by tube agglutination and immunoprecipitation (gel diffusion) tests. Thirty-one of these sera gave suspicious titres and one a positive titre in the tube agglutination test. Six of the rapid slide test positive sera gave positive reactions in immunoprecipitation tests. One dog was identified which was found to have a history very suggestive of B. canis infection. It was judged that 0.3% of sera tested showed serological evidence of B. canis infection. The complexities of the serological diagnosis of B. canis infection was apparent, in particular the tendency to false-positive results in the rapid slide agglutination test.     

Investigations into the prevention of prenatal and lactogenic Toxocara canis infections in puppies by application of moxidectin to the pregnant dog

Aim of the investigation was to examine whether two administrations of moxidectin to pregnant dogs could prevent pre-natal and lactogenic infections of puppies with reactivated Toxocara canis larvae. Four pregnant beagles, infected experimentally with 20 000 embryonated eggs of T. canis, were treated subcutaneously with 1 mg moxidectin per kg body weight on days 40 and 55 of pregnancy (5-13 days before parturition). One further dam and its puppies served as untreated control. Two applications of moxidectin completely prevented pre-natal and lactogenic infections in the puppies. Neither intestinal stages nor somatic larvae were found in the dams or their corresponding puppies. All puppies and dams of the treatment group remained coproscopically negative until 42 days after parturition. The administration of moxidectin did not show any side effects in the dams. None of the puppies of the treated dams showed any pathological abnormalities. In the untreated dam one adult and 26 somatic larvae of T. canis were detected at necropsy. All puppies of the untreated dam showed a patent T. canis infection from day 28 post-natum (p.n.); 296 pre-adult and adult stages of T. canis were spontaneously eliminated and 51 intestinal stages and five somatic larvae of T. canis were recovered at necropsy. In contrast to the puppies of the treated dams all negative control puppies showed blood eosinophilia after parturition and elevated liver enzyme levels.     

Serological responses of dogs to cell wall and internal antigens of Brucella canis (B. canis)

The serological responses of dogs to cell wall and internal antigens of B. canis were studied in experimentally infected specific-pathogen-free (SPF) Beagles. Sera from infected and false positive field dogs also were examined. Cell wall antigens were extracted from B. canis by two procedures that employed either hot phosphate buffered saline (PBS) or sodium desoxycholate (SDC). Agar gel immunodiffusion (AGID) tests employing sera from experimentally infected SPF dogs were used to evaluate antigenic extracts. Extraction with PBS yielded two antigens; SDC extracted an antigen complex and sonication of PBS extracted cells liberated four internal antigens.Sera from field dogs that were negative for B. canis infection in repeated tests often had heterospecific antibodies. Such cross-reactive sere commonly gave “spur” (partial fusion) reactions with a positive reference serum when tested against the SDC cell wall antigen. In addition, false positive dogs did not have antibody to one of the cell wall antigens or to the internal antigens. In contrast, sera from infected field dogs commonly gave “identity” (fusion) reactions in the AGID test with two antigens in the SDC extract, and produced precipitin lines to one to four internal antigens.Examination of a library of sera obtained from experimentally infected SPF dogs over a period spanning 4$\text{1}\text{2}$ years revealed that none of the serodiagnostic tests employed (tube agglutination, slide agglutination, AGID) was accurate during the inital 12 weeks of infection; hemocultures were the most sensitive during this period. Tube and slide agglutination tests were initially sensitive, but they showed a lack of sensitivity and specificity after the bacteremic period ceased, as well as in their failure to exclude false positive reactions in field animals. Immunodiffusion tests that employed SDC or PBS extracts of B. canis cell walls were sensitive and accurate in identifying most infected dogs. After the bacteremia had ceased, however, AGID tests that employed cell wall antigens gave equivocal results. Immunodiffusion tests that employed sonicated (internal) antigens were sensitive shortly after the onset of bacteremia, and they had the advantage of detecting infected animals for at least 6 months following the cessation of bacteremia, a time when other serological tests gave equivocal results.     

Nitroscanate A new broad spectrum anthelmintic against nematodes and cestodes of dogs and cats.

SUMMARY: The efficiency of the broad spectrum anthelmintic nitroscanate against tapeworm and nematode infections of dogs was tested in artificially or naturally infected dogs. The safety of the compound was also evaluated in acute, subacute and chronic toxicity tests. At the recommended single dose rate of 50 mg/kg given to the dogs with food, nitroscanate was 98% efficient against Taenia hydatigena, T. ovis and T. pisiformis. The drug was highly efficient against Echinococcus granulosus only at the dose rate of 200 mg/kg given twice but total elimination of worms was not achieved. Natural infections of Dipylidium caninum, Ancylostoma spp and Uncinaria stenocephala were totally eliminated from all dogs at the dose rate of 25 mg/kg or higher. Nitroscanate was 97% efficient against adult Toxocara canis at a single dose of 50 mg/kg. When the dose was repeated 24 hours later total elimination of both mature worms and immature worms in puppies aged 2 weeks was achieved. The repeated dose of 100 mg/kg removed 98% of early immature worms from puppies aged 3 days. The drug was 100% efficient against adult Toxascaris leonina at a single dose of 50 mg/kg and 90.5% efficiency was achieved against early 4th stage larvae by two doses of 50 mg/kg. Nitroscanate was not efficient against Trichuris vulpis. The drug was efficient for the removal of Toxocara cati and Ancylostoma tubaeforme from cats. Nitroscanate caused no serious symptoms of toxicity at dose rates up to 10,000 mg/kg in single or repeated doses in young or older adult dogs. The drug was safely given to dogs during pregnancy, to young puppies and to cats. The regular use of nitroscanate as a broad spectrum anthelmintic for the prevention and control of parasitic infections of dogs is discussed.     

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.     

Evaluation of recombinant fructose-1,6-bisphosphate aldolase ELISA test for the diagnosis of Schistosoma japonicum in water buffaloes

Fructose-1,6-bisphosphate aldolase (FBPA) is an ubiquitous enzyme essential for glycolysis, gluconeogenesis and the Calvin cycle. It has been demonstrated to induce immune responses and to be useful in the immunodiagnosis of malaria. In this study, FBPA was cloned from the adult worms of Schistosoma japonicum and tested as an antigen for the diagnosis of S. japonicum infection in water buffaloes. Enzyme-linked immunosorbent assay (ELISA) was performed on the sera from 32 infected water buffaloes and 20 negative controls using the recombinant FBPA protein or soluble worm antigen preparation (SWAP) as an antigen. The OD cut-off values were determined to be 0.57 with 100% specificity and 100% sensitivity for the FBPA ELISA and 1.13 with 93.8% specificity and 95.0% sensitivity for the SWAP ELISA. These findings indicate that the recombinant FBPA of S. japonicum should be an useful diagnostic tool for the detection of antibodies against S. japonicum.     

Hair coat contamination with zoonotic helminth eggs of farm and pet dogs and foxes

Infections of dogs with Toxocara canis and Echinococcus multilocularis pose an infection-risk particularly for contact persons. We examined specimens of hair coat and faeces of 124 farm dogs, 118 household dogs, 49 kennel dogs, 15 puppies from two litters, and 46 red foxes. Microscopically identified eggs of Toxocara or taeniids were further investigated by species-specific PCRs. In farm dogs, eggs of E. multilocularis or T. canis were identified in each 2.4% of faecal samples, eggs of T. cati (gastrointestinal passage) in 7.3%, respectively. Household dogs excreted eggs of T. canis (0.8%) and of T. cati (2.5%). In kennel dogs, eggs of T. canis (4.1%), but not of T. cati were detectable. Coat samples contaminated with eggs of Toxocara spp. were found from farm dogs (5.6%), household dogs (1.7%) and kennel dogs (2.0%). Taeniid eggs were isolated from the coat samples from only two farm dogs (1.6%); a molecular species determination was not achieved. In six intrauterinely infected puppies, Toxocara-eggs were found in 17/38 samples taken within six weeks. No intact Toxocara eggs could be isolated from the coat of nine puppies from a second litter 13 days after deworming. Of the 46 red foxes investigated (dissection and faecal samples) 13 (28.3%) were infected with E. multilocularis and 20 (43.5%) with Toxocara. Eggs of taeniids and Toxocara were found in 13% (in three cases confirmed as E. multilocularis) and 21.7%, respectively, of the coat samples. None of the retrieved Toxocara eggs in the coat samples were embryonated. Thus, an infection of humans through the transmission of E. multilocularis eggs after direct contact with dogs or foxes is conceivable, whereas a corresponding infection risk by Toxocara eggs must be critically challenged.     

Periparturient immunosuppression in the bitch and its influence on infection with Toxocara canis

Pregnancy and lactation in dogs induced a marked suppression of their immunological responsiveness as judged by in vitro phytomitogen- and Toxocara canis antigen-induced lymphocyte transformation. In addition, the eosinophilia otherwise seen following infection with T. canis was suppressed during the periparturient period. Coincidental with this periparturient immunosuppression was the occurrence of heavy infections with T. canis in the intestine of lactating bitches and the establishment of such infections perhaps may be related to the suppressed immune reactivity observed. These infections in the bitch appeared to be acquired primarily through the ingestion of immature stages of T. canis passed in the faeces of their puppies. The results demonstrate that the lactating bitch can be a major source of contamination of the environment with eggs of T. canis.     

Efficacy of a milbemycin oxime-praziquantel combination product against adult and immature stages of Toxocara cati in cats and kittens after induced infection

Two studies were performed to examine the efficacy of milbemycin oxime against fourth-stage larvae or adults of Toxocara cati. In the study to determine efficacy against fourth-stage larvae, 20 domestic shorthair cats were inoculated with 500 embryonated eggs. Four weeks after inoculation, the animals were allocated to two groups, and cats in one group were treated with medicated tablets containing 4 mg milbemycin oxime and 10mg praziquantel (MILBEMAX) and cats in the other group with placebo tablets. Seven days after treatment the animals were euthanatized and necropsied for worm counting. The number of worms found was significantly (p=0.0002) lower in cats treated with medicated tablets than in cats treated with placebo tablets. The reduction in the number of worms was 96.53%. In the study to determine efficacy against mature adult worms, 13 kittens were inoculated with T. cati embryonated eggs. On day 45 after inoculation and after the infection had been confirmed through faecal examinations for 11 out of the 13 animals, the 11 infected animals were allocated to two groups and treated as in the first study. Seven days after treatment, all animals were euthanatized and necropsied for worm counting. The number of worms found was significantly (p=0.0043) lower in kittens treated with medicated tablets than in kittens treated with placebo tablets. The reduction in the number of worms was 95.90%. No adverse effects were recorded during either study. It is concluded that the milbemycin oxime-praziquantel tablets that were used are efficacious for the control of T. cati infections in cats.     

Preparation and diagnostic utility of a hemagglutination inhibition test antigen derived from the baculovirus-expressed hemagglutinin-neuraminidase protein gene of Newcastle disease virus

A recombinant hemagglutinin-neuraminidase (rHN) protein from Newcastle disease virus (NDV) with hemagglutination (HA) activity was expressed in Spodoptera frugiperda cells using a baculovirus expression system. The rHN protein extracted from infected cells was used as an antigen in a hemagglutination inhibition (HI) test for the detection and titration of NDV-specific antibodies present in chicken sera. The rHN antigen produced high HA titers of 2¹³ per 25 µL, which were similar to those of the NDV antigen produced using chicken eggs, and it remained stable without significant loss of the HA activity for at least 12 weeks at 4℃. The rHN-based HI assay specifically detected NDV antibodies, but not the sera of other avian pathogens, with a specificity and sensitivity of 100% and 98.0%, respectively, in known positive and negative chicken sera (n = 430). Compared with an NDV-based HI assay, the rHN-based HI assay had a relative sensitivity and specificity of 96.1% and 95.5%, respectively, when applied to field chicken sera. The HI titers of the rHN-based HI assay were highly correlated with those in an NDV-based HI assay (r = 0.927). Overall, these results indicate that rHN protein provides a useful alternative to NDV antigen in HI assays.     

Factors affecting disease manifestation of toxocarosis in humans: Genetics and environment

Toxocara canis is regarded as the main cause of human toxocarosis but the relative contribution of T. cati is probably underestimated; serological and other diagnostic methods used in most studies of this zoonotic disease do not distinguish between the two parasites. The definitive hosts for T. canis are caniidae. Pups generally have higher infection rates than adult animals and are a major source of eggs in the environment. Humans usually acquire T. canis infection by accidental ingestion of embryonated eggs or encapsulated larvae from the environment or contaminated food, such infections may lead to visceral larva migrans (VLM), ocular larva migrans (OLM) or covert toxocarosis (CT). Although a mixed Th1- and Th2-mediated immunological response, particularly with high levels of IgE and eosinophilia is observed, the underlying mechanisms of molecular and immunopathogenesis for the development of the symptomatic syndromes of VLM, OLM, or of asymptomatic CT are largely unclear. Studies have indicated that immunological defences against various infectious diseases may be highly influenced by complex interactions of environmental and host genetic factors e.g. MHC class I and II, also known as human leucocyte antigen (HLA). Toxocara spp. infections are associated with a polarized CD4⁺ Th2 response with high IgE levels and eosinophilia, mediated mainly by HLA class II molecules. Associations have been made between HLA class II and pathological severity and host genetic effects on exposure to infection. Recent research suggests Foxp3⁺ CD4⁺CD25⁺-expressing T regulatory (Treg) cells play a role in regulation of the immunopathology of granulomas in experimental toxocaral granulomatous hepatitis and in enhanced expression of TGF-β1, which is an important factor for the local survival and function of Treg observed during T. canis invasion in the mouse small intestine, liver, muscle, and brain. Since the potential susceptibility loci HLA class II molecules, are considered involved in the regulation of a Th2-dominant immunity which is highly controlled by Foxp3⁺ CD4⁺CD25⁺ Treg cells by stimulation through TGF-β1, which thus provides a beneficial environment to T. canis larvae but severe injuries to local organs. However, TGF-β1 variant Leu10Pro known to be involved in disease severity warrants further elucidation as this too may have a role in the severity of human toxocarosis. Exploration of TGF-β1 polymorphism, Foxp3⁺ CD4⁺CD25⁺ Treg cells, and MHC polymorphisms may allow insight into the contribution made by environmental and genetic factors in influencing disease syndrome type and severity in humans with toxocarosis.     

The migration of larval Toxocara canis in mice II. Post-intestinal migration in primary infections

Following oral infection of NIH mice with Toxocara canis embryonated eggs the L₂ pass the visceral phase of migration during the first week of infection. Larvae reach the liver and lungs and peak in number in these organs 2 and 3 days after infection, respectively. Larvae are then dispersed throughout the body and enter the myotropic—neurotropic phase by the 7th day of infection. Larvae injected directly into the brain are capable of migrating into the viscera and musculature. Considerable pathology occurs due to larval migrations, especially through the liver and lungs, and both acute and chronic disease are recorded. Studies of infections extending over a year show that the number of recoverable larvae declines gradually with periods of stable populations.On Days 3, 4 and 5 after infection, larvae were demonstrable in the faeces of infected mice. Prenatal infection was observed in a third of the offspring of mice infected the same day as conception.     

Vaccination of calves against Taenia saginata infection using a “parasite-free” vaccine

Calves were vaccinated with antigens collected during in vitro cultivation of the larval stages of Taenia ovis, T. hydatigena or T. saginata, and challenged 4 weeks later with 4 000 T. saginata eggs. Calves vaccinated with T. saginata antigen were highly resistant to the challenge infection and those groups vaccinated with T. ovis and T. hydatigena were also significantly resistant.     

Molecular assessment of the transplacental transmission of Toxoplasma gondii,Neospora caninum,Brucella canis and Ehrlichia canis in dogs

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents.     

Evaluation of the efficacy of oxfendazole against larval and adult Toxascaris leonina in the dog

The efficacy of oxfendazole, given at a dose rate of 10 mg/kg for three consecutive days, against adult and larval Toxascaris leonina was determined in four litters of naturally infected adolescent greyhounds. When administered to five dogs 10 weeks after exposure to infectiori, oxfendazole gave an efficacy value of 100 per cent as determined by comparison of the numbers of adult worms expelled and the numbers remaining at subsequent post mortem examination. When medication was given only five weeks after exposure to infection, the number of immature T. leonina in 10 treated pups was found to be reduced by 92-1 per cent as compared with 10 matched, untreated controls. Incidental infections of Toxocara canis and Uncinaria stenocephala were adequately controlled but the treatment was less effective against Trichuris vulpis and Strongyloides species.     

囊虫病猪血清CA效价与虫负荷的相关性及CA与Ab的动态观察

应用ELISA双抗体夹心法检测囊虫病猪血清中的循环抗原(CA),对CA效价与虫负荷的相关性作了初步观察,发现两者密切相关.轻度感染猪血清CA效价≤000,中度为1000～10000,重度为10000～100000.人工感染病猪血清一般在感染后1周检出CA,于感染后4～5周达高峰;抗体一般于感染后2周检出.这种方法可检出的最小虫负荷约为20个/猪