• Journal of Veterinary Clinics
• /
• v.21 no.3
• /
• pp.248-252
• /
• 2004

DNase activity from infective larvae of the parasitic nematode Haemonchus contortus was characterized and compared to that from whole worm. DNase activity from infective larvae was detected throughout pHs 4-10, but high activity was detected under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but was significantly inhibited at pH 7.0. The activity produced DNA, fragments with mixtures of 3'-hydroxyls (OH) and 3'-phosphates (P) at each pH with predominance of 3'-P. A unique DNase activity at 37 kDa was identified from infective larvae on zymograms. The 37 kDa DNase was detected only at pH 5.0, but not at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 37 kDa infective larval DNase resemble those of classic acidic DNases (e.g., DNase II). In contrast, 34, 36 and 38.5 kDa DNase activities were shown to be specific for whole worm. This result demonstrated that DNases in H contortus are regulated during development.

• Korean Journal of Veterinary Research
• /
• v.15 no.1
• /
• pp.51-55
• /
• 1975

In this study two sets of experiments were undertaken. Firstly, the embryonated eggs and first-stage larvae, sprayed in disinfected soil, were exposed to different temperatures. Then, the viabilities of them were weekly determined by inoculating the soil to earthworms, Eisenia foetida. Secondly, the infective larvae, seperated from the earthworms and suspended in tap water, were also exposed to different temperatures and their viabilities were checked microscopically at weekly intervals. The results were summerized as follows: 1. The maximum longevities of embryonated eggs and first-stage larvae were determined as 1 week at $35^{\circ}C$, over 36 weeks at $25^{\circ}C$, $15^{\circ}C$, and $5^{\circ}C$, 32 weeks at $-5^{\circ}C$, and under 1 week at $-15^{\circ}C$. 2. The mean numbers of infective larvae detected from the test earthworms were greatest at $5^{\circ}C$, and decreased with rise or fall of the temperature. 3. Infective larvae freed from the intermediate host were able to survive for 2 weeks at $25^{\circ}C$ and 3 weeks at $15^{\circ}C$. However, they lost their viabilities in a week at $35^{\circ}C$, $5^{\circ}C$, $-5^{\circ}C$, and $-15^{\circ}C$. 4. The number of living infective larvae at $15^{\circ}C$ was greater than at $25^{\circ}C$.

• Korean Journal of Veterinary Research
• /
• v.11 no.1
• /
• pp.1-39
• /
• 1971

The purpose of present study was to investigate the possibility to immunize guineapigs and swine against Metastrongylus apri infection by the administration of irradiated infective larvae. Four main experiments were undertaken. Firstly, three groups of infective larvae irradiated at $3{\times}10^4r$, $4{\times}10^4r$, and $5{\times}10^4r$ respectively were inoculated to guineapigs and their immunogenic effects were examined from the clinical, anatomical, and serological viewpoints to decide the optimal dose of X-ray for the atenuation of them. Secondly, the migratory behavior of the larvae irradiated at the optimal dose was compared with that of normal infective larvae. Thirdly, pigs were inoculated with each 5,000 infective larvae irradiated at the optimal dose and the clinical, anatomical and serological responses of them before and after challenge were examined. Fourthly, the heated extract of adult M. apri which had been used as an antigen in the serological examinations was analyzed and compared with that of adult Ascaris suis and of adult Trichuris suis by immunoelectrophoretic method. The results obtained are summerized as follows: 1) The optimal dose of X-ray for the atenuation of the infective larvae which can minimize the pathogenecity but keep the antigenecity of the infective larvae was $5{\times}10^4r$. 2) Guineapigs could become completely resistant to subsequent challenge infection by the administration of 1,000 infective larvae irradiated at $5{\times}10^4r$, without showing any symptom of disease before and after the challenge. 3) There were some indication that guineapigs could acquire complete immunity after they overcome the infection with normal infective larvae. 4) It was shown that, in guineapigs, the $5{\times}10^4r$-irradiated larvae can migrate to the large intestine and mesenteric lymph node within a day, where they stay for as long as 16 days to stimulate the host's immunity. 5) It also was shown that, in guineapigs, the normal infective larvae challenged to resistant guineapigs can migrate to the large intestine and mesenteric lymph node, where they are affected by the immune mechanism of host within 10 days without further migration. 6) Pigs could become partially resistant to subsequant challenge by the administration of 5,000 infective larve irradiated at $5{\times}10^4r$; no clinical symptom occurred after the administration, but milder symptoms of parasitic bronchitis were observable after the challenge infection and fewer number of worms were detected from the lungs at autopsy compared with severe symptoms and much number of worms in control pigs. 7) It was shown that, in pigs, a few of the $5{\times}10^4r$-irradiated larvae can migrate to the lungs, where they stay for as long as 104 days in stunted and sterile states; their body-lengths were short and their uteri developted no eggs. 8) There was evidence that the male larvae were more susceptible to X-ray than the female larvae. 9) Antibodies relating to the administration with $5{\times}10^4r$-irradiated or normal larvae were detected from the sera of both guineapigs and pigs by means of indirect haemagglutination and agar diffusion precipitin tests. Relatively higher antibody titers were recorded by the former test, but precipitin bands were demonstrable only when the positive sera were concentrated in one tenth of original volume in the later one. 10) The antibody titers of pig sera began to rose on 14 days, kept their peak during the period from 14 th day to 21st day, and fell to a low level on 28 days after the administration of $5{\times}10^4r$ or normal infective larvae. 11) A slight increase in gamma globublin of the pig sera occurred following the administration. The gamma globulin level showed a tendency to fluctuate in acordance with the antibody level. 12) A marked eosionophilia occurred in pigs on 7 or 14 days following the administration. The eosinophil count showed the same tendency to fluctuate as the gamma globulin did. 13) It was shown that the serum antibodies detected by the heated extract of adult Metastrongylus apri react crossly with the heated extract of adult Ascaris suis but not with that of adult Trichuris suis in indirect haemagglutination and agar diffusion preciption reactions. 14) The heated extract of adult Metastrongylus apri could he divided into 9 antigenic components by immunoelectrophoresis, one (arc 4) of which was shown to be common to both extracts of adult Ascaris suis and adult Trichuris suis, and the other one (arc 9) to only the extract of adult Ascaris suis.

• Korean Journal of Veterinary Research
• /
• v.11 no.2
• /
• pp.157-161
• /
• 1971

This experiment was undertaken in order to investigate whether chicken can serve as a carrier for swine lungworm, Metastrongylus apri. The results abtained are summerized as follows: All chickens which were fed the infective larvae of M. apri, in the form of infected earthworm (Eisenia foctida) excreted appreximately 4.4 to 5.4 percent of the infective larvae in their feces between 2 to 10 hours after dosing. The larval excretion was marked especially during the first 2 hours. A few (0.10~0.12%) larvae were detected from the intestinal wall of chickens by pepsin-digestion method on a day after dosing, but no further migration or development of the larvae was observed. The larvae which had passed through the digestive tract of chickens retained their viability and produced characteristic lungworm disease when inoculated to guineapigs. From these results, it may he concluded that the chicken can serve as a carrier for the infective larvae of M. apri.

• Journal of the korean veterinary medical association
• /
• v.21 no.1
• /
• pp.43-47
• /
• 1985

Striking effects of hydrocortisone were observed on the behaviour of Toxocara canis larvae in 2 months old dogs. Four cross-breed dogs were used in this studies and each dog was inoculated 3000 infective eggs. The larvae were recovered from 2 dogs (contro

• Journal of the korean veterinary medical association
• /
• v.21 no.10
• /
• pp.612-616
• /
• 1985

A prominent effect of prolactin was observed on the migration behaviour of T.canis larvae in 2 months old puppies, Four cross- breed puppies were used in this studies and each puppy was infected with 3000 infective eggs. One of 4 puppies used for control

• Parasites, Hosts and Diseases
• /
• v.36 no.1
• /
• pp.15-22
• /
• 1998

An in uitro culture technique was established for harvesting Strongwloides venezuelensis free-living infective larvae using a nutrient broth medium as a substitute for rat-feces in polyvinyl culture bags ($10{\;}{\times}{\;}12{\;}cm$). The egg hatch rate V) in sterile saline at different incubation temperatures (X) was expressed as the quadratic function, Y = $-0.192X^2$ + 8.673x - 19.550 (r = 0.901). The highest (100%) egghatch rate was observed at $25^{\circ}C$. A significant difference (p<0.05) in development rate W) of free-living infective larvae was observed between different concentrations of nutrient broth (X) which was highest (20.6%1 in 0.12% nutrient broth concentrations, incubated at $20^{\circ}C$ for 5 days [Y = $-864.032X^2$ + 245.995X- 0.560 (r = 0.875)]. Yields (Y) of infective larvae were observed relatively high when the culture medium was incubated at higher temperatures (X) which peaked at $25^{\circ}C$ (20.0%) than at lower temperatures. $15^{\circ}C$M (10.9%) and $20^{\circ}C$ (18.1%) [Y = $-0.189X^2$ + 8.387x- 72.795 (r = 0.981)]. The period W) required for the development of infective larvae decreased with higher incubation temperatures (X) [Y = $0.035X^2$ - 2.025X + 32.375 (r = 0.995)] The highest yield (19.2%) of infective larvae was obtained from culture bag inoculated with 15.000 eggs than with below and over 15,000 eggs in 0.12% nutrient broth and incubated at $25^{\circ}C$ for 4 days. The newly adapted culture method (from egg to third-stage larva) may be useful as a bio-bar/bioassay system for screening new chemical products, anthelmintics and pesticides, as well as for parasito immunological studies with Strongwloides species.

• The Korean Journal of Soil Zoology
• /
• v.5 no.2
• /
• pp.97-100
• /
• 2000

We report here an improved collecting method of the infective juveniles multiplied in the host insect with the entomopathogenic nematodes, Steinernema carpocapsae Weiser. The specific characteristics of this method involved the opening of the host insect hemocoel after the population of their infective juveniles reached at the maximum (6 days at 25$\^{C}$ after nematode treatment to nonimmunized host insects) to facilitate the escape of the multiplied nematodes. It also used 'Baermann funnel'method to select the infective juveniles effectively. This improved 'Baermann funnel'method was compared with a traditional collecting method, which was characterized with a combination of untreated host insects and 'White trap'collecting method, in both yield and pathogenicity of the collected infective juveniles to the fifth instar larvae of beet armyworm, Spodoptera exigua (H bner). More than 95% of the nematode populations collected by the two methods represented the morphological infective juveniles. To prove the nematodes to be infective juveniles functionally, pathogenicity and infective activity were compared in the nematodes collected by the two methods. They were not different in both pathogenicities and infective activities which were measured by the numbers of nematodes penetrated into the hemocoel of the insect hosts after exposure for the specific times to the same dote of infective juveniles. Significant difference between two collecting methods was found in the total yields of the infective juveniles per host insect About 50,000 infective juveniles per infected fifth instar larva of S. exigua after 6 day incubation at 25$\^{C}$ were collected only for 2 days by the improved 'Baermann funnel'method, while about 20,000 infective juveniles per host were collected for 10 days by the classical 'White trap'method.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.6
• /
• pp.968-978
• /
• 2007

A pathogenic nematode, Butlerius sp., was isolated from Oriental beetle, Blitopertha orientalis. The infective juveniles exhibited dose-as well as time-dependent entomopathogenicity on the larvae of B. orientalis. Two bacterial species, Providencia vermicola (KACC 91278) and Flavobacterium sp. (KACC 91279), were isolated from the infective juveniles and identified. P. vermicola outnumbered Flavobacterium sp. in the nematode host, in which the colony density of P. vermicola was found to be 21 times higher than that of Flavobacterium sp. However, when the two bacterial species were cocultured in culture media without the nematode host, they showed similar growth rates. Both bacteria induced significant entomopathogenicity against Spodoptera exigua larvae infesting economically important vegetable crops, where P. vermicola was more potent than Flavobacterium sp.

• The Korean Journal of Soil Zoology
• /
• v.8 no.1_2
• /
• pp.17-22
• /
• 2003

An endemic entomopathogenic nematode, Heterorhabditis megidis, was evaluated by its control efficacies against housefly, Musca domestica, and flower beetle, Gametis jucunda. In Petri-dish assay, the pathogenicity of H. megidis showed 456.4 infective juveniles/larva (IJs/larva) in median lethality (LC$_{50}$) against the second instar larvae of M. domestica and 238.9 IJs/larva against the second instar larvae of G. jucunda. This was contrasted with those of the other well-known entomopathogenic nematode, Steinernema carpocapsae, which showed 115.9 IJs/larva against M. domestica and 388.6 IJs/larva against G. jucunda. In field experiment, H. megidis were applied per square meter of pork farm with 1,000,000 IJs of H. megidis or apple orchard with 370,000 IJs, which were infested with M. domestica or G. jucunda, respectively. H. megidis showed 56.9% and 57.3% of control efficacies against M. domestica and G. jucunda, respectively. These results suggest a promising control technique in the field using H. megidis against M. domestica and G. jucunda.a.