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Pathogenicity of Entomopathogenic Nematode, Steinernema carpocapsae Pocheon Strain Against Anomia mesogona and Anomis commoda(Lepidoptera: Noctuidae) (무궁화잎밤나방(Anomis mesogona)과 큰붉은잎밤나방(Anomis commoda)에 대한 Steinernema carpocapsae 포천 계통의 병원성)

  • Kim Hyeong-Hwan;Park Hyung-Soon;Cho Yoon-Jin;Lee Dong-Woon;Choo Ho-Yul;Goo Kwan-Hyo
    • Asian Journal of Turfgrass Science
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    • v.19 no.1
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    • pp.17-25
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    • 2005
  • Pathogenicity of entomopathogenic nematde, Steinernema carpocapsae Pocheon strain (ScP) was evaluated against different larval stages(2nd, 3-4th and 5th) of Anomis commode and Anomis mesogona(Lepidoptera: Noctuidae) in petri dish and pot. The $LC_{50}$ values were increased in proportion to larval stage of A. commoda and A. mesogona. $LC_{50}$ value of ScP against 2nd instar of A. commoda and A. mesogona was 9.7 and 4.5, respectively. The 2nd instar of both species was also susceptible to ScP in pot test. That is, mortality of 2nd instar of A. commode and A. mesogona was higher representing $72.5\%\;and\;87.5\%$ 5 days later after treatment, respectively, when ScP was applied at the ,ate of 90,000 infective juveniles(Ijs) per $pot(=1\times10^9\;Ijs/ha)$. However, susceptibility was decreased from 3rd instar. Mortality of 5th instar of A. commoda and A. mesogona was $5.0\%\;and\;10.0\%$, respectively, at the rate of 90,000 Ijs/pot. When ScP was applied into pot including mixed larval stages from 2nd to 5th instar(10 larvae far 2nd instar, 10 larvae for 3rd - 4th instars, and 10 larvae for 5th instar) at the rate of 90,000 Ijs/pot, mortality of A. commoda and A. mesogona was $69.2\%\;and\;50.0\%$, respectively.

Influence of Dietary Supplementation of Condensed Tannins through Leaf Meal Mixture on Intake, Nutrient Utilization and Performance of Haemonchus contortus Infected Sheep

  • Pathak, A.K.;Dutta, Narayan;Banerjee, P.S.;Pattanaik, A.K.;Sharma, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1446-1458
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    • 2013
  • The study assessed the effect of dietary supplementation of leaf meal mixture (LMM) containing condensed tannins (CT) on feed intake, nutrient utilization and performance of sheep infected with Haemonchus contortus. Eighteen adult sheep of similar age and body weight ($25.03{\pm}1.52$) were included in this study and out of these, 12 sheep were infected with single dose of infective third stage larvae of H. contortus at 2,000 larvae per sheep. The experimental sheep were allocated in three different groups' i.e. negative control (NC; no infection), control (C; H. contortus infected) and treatment (T; H. contortus infected+CT at 1.5% of the DM through LMM) and the experiment was conducted for a period of 90 d. The intake of dry matter (DM), organic matter (OM) and digestibility of DM, OM, neutral detergent fibre (NDF) and acid detergent fibre (ADF) were comparable among three animal groups. However, digestibility of crude protein (CP) and ether extract (EE) were significantly (p<0.05) higher in NC group as compared to both C and T groups. Nitrogen (N) retention (g/d or % of N intake) was significantly (p = 0.038) lower in C group as compared to T and NC groups. Daily intake (g/kg $W^{0.75}$) of digestible crude protein (DCP), digestible organic matter (DOM) and total digestible nutrient (TDN) did not differ significantly (p<0.05) in the three groups. Haemoglobin (Hb) and packed cell volume (PCV) were significantly (p<0.001) higher in treatment group as compared to control. The level of Hb and PCV reduced (p<0.001) after 30 days of experimental feeding. CT significantly (p<0.001) reduced serum urea in T group as compared to NC and C groups. Serum proteins differed significantly (p<0.01) among the three groups. The activity of serum enzymes AST, ALT, ALP and LDH were also statistically non significant (p<0.05) among treatments. The weight of abomasal lymph nodes (ALN) in T group was higher (p<0.05) than in C group. Treatment group had lower (p<0.05) total worms and fecal egg count compared to control group. It may be concluded that dietary supplementation of CT through LMM significantly improved the N retention, and inhibited the different developmental stages of Haemonchus contortus in experimental sheep.

Isolation and Identifieation of Entomopathogenic Nematodes from Soil and Insect (토양과 곤충 사체로부터 곤충병원성 선충의 분리 및 동정)

  • 한상미;한명세
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.321-330
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    • 1999
  • Nematodes were isolated using silkwom trap through the investigation of 100 soil samples from various biotopes in Korea. The 30 nematode strains from soil and dead insects by the pathogenicity aganinst silkworms (Bombyx mori mori) and insect pests of Calliphora vomitoria, Pseufazetia separata, Palomena angulosa, and Melolontha incana. Mortailty of the silkworm larvae and pupae were as high as 100% by nematode infection, those of insect of pests were varied from 20 to 100%. The 30 strains of entemopathogenic nematodes were classified into five groups of Rhabditidae, Diplogatroidae, Heterorhabitidae, Steinernematidae, and Tylenchida by morphological criteria. The genetic relationships among the 30 nematode strains were analyzed by various RAPD bands with twenty primers. The 30 nematode strains were classified into six major subgroups on the basis of the genetic similarity coefficient of 0.853. The grouping by RAPD was agree with those of morphological taxa in discrimination of the higher group, however, was not completely agree in the subgroup. The family Steinernematidae belong to Rhabditida was clarified as closer to the Tylenchida, rather than the other Rhabditida of Heterorhabitidae, Rhabditidae, and Diplogatroidae in genetic distance valule. From the result of the morphological classification and RAPD of the genomic DNA showed that genetic relationship analysis furnish infurmation on phylogenetic classification and relationships of entomopathogenic nematodes. The application of genetic similarity will overcome the limitation of taxonomy and classification of morphologically simple nematode. Several primers were confirmed those utility of identification for individual nematode strains, the methods of molecular genetics secured the simplicity, rapidity and accuracy on the selection of entomopathogenic nematodes.

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Leaf Spray Control Efficacy of the Entomopathogenic Nematode, Steinernema carpocapsae Weiser, Supplemented with the Selected Antidesiccant, Keltrol-F, on the Beet Armyworm, Spodoptera exigua(Hubner) (선발 내건제 Keltrol-F를 이용한 곤충병원선충(Steinernema carpocapsae Weiser)의 파밤나방(Spodoptera exigua(Hubner))에 대한 엽면살포 방제 효과)

  • 이성섭;김용균;한상찬
    • Korean journal of applied entomology
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    • v.39 no.3
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    • pp.199-205
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    • 2000
  • The field control efficacy of entomopathogenic nematode, Steinernema carpocapsae Weiser, was evaluated on the beet armyworm, Spodoptera exigua (Hiibner). The insect pest has been known to be a defoliator at the aerial part of the crop and difficult to be controlled effectively with most commercial chemical insecticides due to its insecticide resistance. To overcome the susceptibility of the nematodes to desiccation when they were applied by leaf spray in field condition, we screened several commercial antidesiccants (alkyl glucoside, CMC, glycerol, Keltrol-F, Kunipia-G, and Laponite LXG) optimal for survival of the nematodes. Keltrol-F (0.1 %) was selected as a candidate supplement for field application of the nematodes. Leaf spray of the nematodes at 5,000 infective juveniles/ml of distilled water containing 0.1 % Keltrol-F resulted in 87.7% control efficacy on the 3rd instar larvae of Sp. exigua.

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Construction of a Transposon-mediated Baculovirus Vector Hanpvid and a New Cell Line for Expressing Barnase

  • Qin, Qin;Liu, Ying-Le;Zhu, Ying;Li, Shun-Yi;Qi, Yi-Peng
    • BMB Reports
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    • v.38 no.1
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    • pp.41-48
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    • 2005
  • In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AM1-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM1 and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages ($LD_{50}$) and the lethal time(s) ($LT_{50}$) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

EFFECT OF EXPERIMENTAL Haemonchus contortus INFECTION ON HEMOGLOBIN CONCENTRATION AND PACKED CELL VOLUME OF DOES

  • Howlader, M.M.R.;Capitan, S.S.;Eduardo, S.L.;Roxas, N.P.;Sevilla, C.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.5
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    • pp.597-601
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    • 1996
  • Twelve Philippine does of 3.5-5.0 years old were used in this study. They were divided into three groups, $T_1$, $T_2$ and $T_3$. Four animals were randomly allocated to each group. Before infection, they were dewormed and housed in individual pens with concrete floors. They were provided with a uniform management. They were infected orally with a single dose of three levels (0, 15,000 and 30,000) of infective Haemonchus contortus larvae. Blood samples were collected from the jugular vein of each animal at fortnightly intervals for 17 fortnights. Hemoglobin (Hb) concentration was determined using hemoglobinometer and packed cell volume (PCV) by microhematocrit methods. Animals in infected groups showed significantly (p < 0.05) lower hemoglobin values than the control except during fortnight 1 for group 2 and fortnights 1, 2, 5, 6 and 7 for group 3. Hemoglobin concentration did not significantly (p > 0.05) differ between the animals in infected groups throughout the sampling period except fortnight 2. The PCV values of animals in infected groups were significantly (p < 0.05) lower than the control for most of the sampling periods. The PCV values of animals in group 2 did not significantly (p > 0.05) differ from group 3.

Studies on Ancylostomiasis I. An Experimental Study on Hookworm Infection and Anemia (구충증(鉤蟲症)에 관(關)한 연구(硏究) 제1편(第1篇) 구충(鉤蟲)의 감염(感染) 및 구충성빈혈(鉤蟲性貧血)에 관(關)한 고찰(考奈))

  • Lee, Mun-Ho;Kim, Dong-Jip;Lee, Jang-Kyu;Seo, Byong-Sul;Lee, Soon-Hyung
    • The Korean Journal of Nuclear Medicine
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    • v.1 no.1
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    • pp.55-66
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    • 1967
  • In view of its prevalence in the Far East area, a more detailed knowledge on the hookworm infection is one of the very important medical problems. The present study was aimed to; determine the infectivity of the artificially hatched ancylostoma duodenale larvae in man after its oral administration, evaluate the clinical symptomatology of such infection, determine the date of first appearance of the ova in the stool, calculate the blood loss per worm per day, assess the relation-ships between the ova count, infectivity(worm load), blood loss and severity of anemia. An erythrokinetic study was also done to analyse the characteristics of hookworm anemia by means of $^{59}Fe\;and\;^{51}Cr$. Materials and Methods Ten healthy male volunteers(doctors, medical students and laboratory technicians) with the ages ranging from 21 to 40 years were selected as the experimental materials. They had no history of hookworm infection for preceding several years, and care was taken not to be exposed to reinfection. A baseline study including a through physical examinations and laboratory investigations such as complete blood counts, stool examination and estimation of the serum iron levels was done, and a vermifuge, bephenium hydroxynaphoate, was given 10 days prior to the main experiment. The ancylostoma duodenale filariform larvae were obtained in the following manner; The pure ancylostoma duodenale ova were obtained from the hookworm anemia patients and a modified filter paper method was adopted to harvest larger number of infective larvae, which were washed several times with saline. The actively moving mature larvae were put into the gelatine capsules, 150 in each, and were given to the volunteers in the fasting state with 300ml. of water. The volunteers were previously treated with intramuscular injection of 15mg. of chlorpromazine in order to prevent the eventual nausea and vomiting after the larvae intake. The clinical symptoms and signs mainly of the respiratory and gastrointestinal tracts, appearance of the ova and occult blood in the stool etc. were checked every day for the first 20 days and then twice weekly until the end of the experiment, which usually lasted for about 3 months. Roentgenological survey of the lungs was also done. The hematological changes such as the red blood cell, white blood cell and eosinophil cell counts, hemoglobin content and serum iron levels were studied. The appearance of the ova in the stool was examined by the formalin ether method and the ova were counted in triplicate on two successive days using the Stoll's dilution method. The ferrokinetic data were calculated by the modified Huff's method and the apparent half survival time of the red blood cells by the modified Gray's method. The isotopes were simultaneously tagged and injected intravenously, and then the stool and blood samples were collected as was described by Roche et al., namely, three separate 4-day stool samples with the blood sample drawing before each 4-day stool collection. The radio-activities of the stools ashfied and the blood were separately measured by the pulse-height analyser. The daily blood loss was calculated with the following formula; daily blood loss in $ml.=\frac{cpm/g\;stool{\times}weight\;in\;g\;of\;4-day\;stool}{cpm/ml\;blood{\times}4}$ The average of these three 4-day periods was given as the daily blood loss in each patient. The blood loss per day per worm was calculated by simply dividing the daily blood loss by the number of the hookworm recovered after the vermifuge given twice a week at the termination of the experiment. The iron loss in mg. through the gastrointestinal tract was estimated with the daily iron loss in $mg=\frac{g\;Hgb/100ml{\times}ml\;daily\;blood\;loss{\times}3.40}{100}$ 3.40=mg of iron per g Hgb following formula; Results 1. The respiratory symptoms such as cough and sputum were noted in almost all cases within a week after the infection, which lasted about 2 weeks. The roentgenological findings of the chest were essentially normal. A moderate degree of febril reaction appeared within 2 weeks with a duration of 3 or 4 days. 2. The gastrointestinal symptoms such as nausea, epigastric fullness, abdominal pain and loose bowel appeared in all cases immediately after the larvae intake. 3. The reduction of the red blood cell count was not remarkable, however, the hemoglobin content and especially the serum iron level showed the steady decreases until the end of the experiment. 4. The white blood cells and eosinophil cells, on the contrary, showed increases in parallel and reached peaks in 20 to 30 days after the infection. A small secondary rise was noted in 2 months. 5. The ova first appeared in the stool in 40. 1 days after the infection, ranging from 29 to 51 days, during which the occult blood reaction of the stool became also positive in almost cases. 6. The number of ova recovered per day was 164, 320 on the average, ranging from 89,500 to 253,800. The number of the worm evacuated by vermifuge was in rough correlation with the number of ova recovered. 7. The infectivity of ancylostoma duodenale was 14% on the average, ranging from 7.3 to 20.0%, which is relatively lower than those reported by other workers. 8. The mean fecal blood loss was 5.78ml. per day, with a range of from 2.6 to 11.7ml., and the mean blood loss per worm per day was 0.30ml., with a range of from 0.13 to 0.73ml., which is in rough coincidence with those reported by other authors. There appeared to exist, however, no correlation between the blood loss and the number of ova recovered. 9. The mean fecal iron loss was 2.02mg. per day, with a range of from 1.20 to 3.89mg., which is less than those appeared in the literature. 10. The mean plasma iron disappearance rate was 0.80hr., with a range of from 0.62 to 0.95hr., namely, a slight accerelation. 11. The hookworm anemia appeared to be iron deficiency in origin caused by continuous intestinal blood loss.

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Effect of Entomopathogenic Nematodes on Egg Mass Formation by the Northern Root-knot Nematode, Meloidogyne hapia (곤충병원성 선충이 당근뿌리혹선충의 난낭 형성에 미치는 영향)

  • 김형환;추호렬;조명래;전흥용;임명순
    • Korean journal of applied entomology
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    • v.41 no.3
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    • pp.225-231
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    • 2002
  • The entomopathogenic nematodes, Steinernema carpocapsae All strain (ScA), S.glaseri NC strain (SgN) and H. bacteriophora NC 1 strain (HbN), were evaluated for the effects on egg mass formation by the northern root-knot nematode, Meloidogyne hapla in pot experiment using tomato. In the first experiment, 2.5$\times$10$^{5}$ infective juveniles (Ijs) of entomopathogenic nematodes were inoculated to 100 g of the soil infected with ca. 450 Ijs of M. hapla/100 ㎤ in 150 $mell$ container. The number of egg mass was significantly decreased to 9.4-36.5 in ScA, to 5.7-24.7 in SgN and to 11.2-16.0 in HbN treatments compared with 62.5 in M.hapla alone. In the second experiment, ScA and S.carpocapsae Pocheon strain (ScP) and SgN and S.glaseri Dongrae strain (SgD) were treated to 350 g of the soil infected with 100, 200 M.hapla larvae/100 ㎤ in 450 $mell$ container The entomopathogenic nematodes were inoculated at the rate of 2,020 Ijs and 1.6$\times$105 Ijs in 350 g soil. The number of egg mass of M.hapla were significantly decreased in the entomopathogenic nematode treatments compared with M.hapla alone although no differences were observed among Steinernema species, strains, or infection concentrations. Treatments of entomopathogenic nematodes 3 days before M.hapla inoculation were more effective on reduction of egg mass formation than those of 3 days after M.hapla treatments. Growth of tomato was not affected by entomopathogenic nematode treatments.

Evaluation of Entomopathogenic Nematodes against Armyworm, Pseudaletia separata on Tall Fescue, Festuca arundinacea (톨페스큐에서 곤충병원성선충의 멸강나방에 대한 효과 검정)

  • Jung, Young Hak;Kim, Jong Ju;You, Eun Ju;Lee, Chae Min;Choo, Ho Yul;Lee, Dong Woon
    • Weed & Turfgrass Science
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    • v.2 no.3
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    • pp.312-317
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    • 2013
  • The armyworm, Pseudaletia separata was occurred suddenly in a golf club in Namhae, Gyeongnam province in 2013. Thus, pathogenicity of seven species of entomopathogenic nematodes (Steinernema carpocapsae GSN1 strain, S. glaseri Dongrae strain, S. longicaudum Nonsan strain, S. monticolum Jiri strain and S. siamkayai, Heterorhabditis sp. Gyeongsan strain, and H. bacteriophora Hamyang strain) was evaluated against armyworm on tall fescue in pot and golf course to control this pest environmentally friendly. The pathogenicity against P. separata larvae was significantly different depending on nematode species. The corrected mortality of fifth instar of P. separata was 100% in the treatment with S. carpocapsae GSN1 in 7 days in tall fescue pot. However, there was not different in the mortality of fifth instar of P. separata (80 to 100%) at the rate of 385 to 6,160 infective juveniles (Ijs) ($=2.5{\times}10^2$ to $4{\times}10^5Ijs/m^2$) of S. carpocapsae GSN1 strain in each pot. Corrected mortality of P. separata was 65 and 60% at the rate of $10^5Ijs/m^2$ of S. carpocapsae GSN1 and S. longicaudum Nonsan strain, respectively in the tall fescue of golf course.

Pathogenicity of Entomopathogenic Nematode, Steinernema carpocapsae GSN1 Strain (Rhabditida: Steinernematidae) against Tebenna issikii (Lepidoptera: Choreutidae) (우엉뭉뚝날개나방(Tebenna issikii)에 대한 Steinernema carpocapsae GSN1 계통의 병원성)

  • Kim, Hyeong-Hwan;Han, Gun-Yeong;Choo, Ho-Yul;Lee, Sang-Myeong;Lee, Dong-Woon
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.313-318
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    • 2007
  • Entomopathogenic nematode, Steinernema carpocapsae GSN1 strain was evaluated for the environmentally sound control of Tebenna issikii (Lepidoptera: Choreutidae) in the laboratory. The corrected mortality of Tebenna issikii larvae was 100% at the 40 infective juveniles (Ijs)/larva 3 days after treatment with S. carpocapsae GSN1 strain in Petri dish. $LC_{50}$ value of S. carpocapsae GSN1 strain against Tebenna issikii was 5.7 Ijs. The mean penetration numbers of Ijs of S. carpocapsae GSN1 strain at the 5, 10, 20, 40 and 80 Ijs/larva in a Tebenna issikii larva were 1.4, 1.4, 3.2, 5.6 and 11.9 Ijs/larva, respectively. However penetration rate of Ijs of S. carpocapsae GSN1 strain at 5 Ijs/larva was the highest among other nematode concentrations. Progeny of S. carpocapsae GSN1 strain in a Tebenna issikii larva was higher with increasing nematode concentration.