• Title/Summary/Keyword: First larval stage

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Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model

  • Wang, Sibo;Yang, Tao;Zhang, Xuyong;Xia, Jie;Guo, Jun;Wang, Xiaoyi;Hou, Jixue;Zhang, Hongwei;Chen, Xueling;Wu, Xiangwei
    • Parasites, Hosts and Diseases
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    • v.54 no.3
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    • pp.291-299
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    • 2016
  • Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

Detection of Gnathostoma spinigerum Third-Stage Larvae in Snakeheads Purchased from a Central Part of Myanmar

  • Jung, Bong-Kwang;Lee, Jin-Ju;Pyo, Kyoung-Ho;Kim, Hyeong-Jin;Jeong, Hoo-Gn;Yoon, Cheong-Ha;Lee, Soon-Hyung;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.46 no.4
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    • pp.285-288
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    • 2008
  • To examine the infection status of freshwater fish with Gnathostoma spp. larvae in Myanmar, we purchased 15 snakeheads, Channa striatus, from a local market in a suburban area of Naypyidaw, the new capital city. Two larval gnathostomes were collected using an artificial digestion technique, and observed by a light microscope and a scanning electron microscope. The size of an intact larva was 2.65 mm long and 0.32 mm wide. The characteristic morphology of the larvae included the presence of a long esophagus (0.80 mm long), 2 pairs of cervical sacs (0.43 mm long), and a characteristic head bulb with 4 rows of hooklets. The number of hooklets in the 1st, 2nd, 3rd, and 4th row was 45, 48, 50, and 52, respectively. Based on these morphological characters, the larvae were identified as the advanced 3rd-stage larvae of Gnathostoma spinigerum. This is the first report of detection of G. spinigerum 3rd-stage larvae in the central part of Myanmar. Our study suggests that intake of raw meat of snakehead fish in Myanmar may result in human gnathostomiasis.

The Distribution Pattern of Eggs and Larval Survival Rate of Phyllocnistis citrella (Lepidoptera: Gracillaridae) on the Leaves of Shoots in a Citrus Orchard in Jeju (제주 감귤원에서 귤굴나방 알의 신초 잎 위치별 분포와 유충의 생존률)

  • Hyun, Seung Young;Kim, Su Bin;Kim, Dong-Soon
    • Korean journal of applied entomology
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    • v.58 no.3
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    • pp.175-182
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    • 2019
  • The citrus leafminer, Phyllocnistis citrella (Lepidoptera: Gracillaridae), is an important pest that decreases the productivity of citrus trees by attacking citrus leaves, and especially it causes a severe damage during the young growth stage. In this study, we investigated the distribution of eggs on the leaves of shoots according to the position and the survival rate of this pest in the field condition. The occurrence patterns of P. citrella eggs in citrus orchards were largely affected by the occurrence of shoots. Even if the adults were still being attracted in the pheromone traps, the eggs were not observed on leaves when the growth of shoots was stopped. The density of P. citrella eggs in the citrus shoots peaked at the fifth or sixth leaf from the terminal leaf, and over 90% of the eggs were found in the first to eighth leaves as a whole. In 2015 and 2016, the average survival rate of larvae surviving to pupa was 1.4%, and the rate of survival to adults was 0.2%, indicating that most of them died. Furthermore, we have discussed the management strategy of P. citrella considering the distribution of eggs and the larval survival rate in the citrus shoots.

Angiostrongylus cantonensis: Scanning Electron Microscopic Observations on The Cuticle of Moulting Larvae

  • Zeng, Xin;Wei, Jie;Wang, Juan;Wu, Feng;Fung, Feng;Wu, Xiaoying;Sun, Xi;Zheng, Huanqing;Lv, Zhiyue;Wu, Zhongdao
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.633-636
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    • 2013
  • Angiostrongylus cantonensis is a parasitic nematode that needs to develop in different hosts in different larval stages. Freshwater snails, such as Pomacea canaliculata, are the intermediate host, and rats are the definitive host. Periodic shedding of the cuticle (moulting) is an important biological process for the survival and development of the parasite in the intermediate and definitive hosts. However, there are few studies on the cuticle alterations between different stages of this parasite. In this study, we observed the ultrastructural appearance and changes of the cuticle of the 2nd/3rd stage larvae (L2/L3) and the 3rd/4th stage larvae (L3/L4) using a scanning electron microscope. We also first divided L2/L3 into late L2 and early L3. The late L2 lacked alae, but possessed a pull-chain-like fissure. Irregular alignment of spherical particles on the cuticle were noted compared to the L3. Alae appeared in the early L3. The old cuticle turned into a thin filmlike structure which adhered to the new cuticle, and spherical particles were seen regularly arranged on the surface of this structure. Regular rectangular cavities were found on the surface of L3/L4. The caudal structure of L3/L4 was much larger than that of L3, but caudal inflation, such as seen in L4, was not observed. These results are the first to reveal the ultrastructural changes of the cuticle of A. cantonensis before and after moulting of L2/L3 and L3/L4.

Adaptive Significance of Changes in the Biochemical Constituents of Haemolymph to Imposed Thermal Stress on Seleted Races of the Silkworm, Bombyx mori L.

  • Malik Firdose Ahmad;Reddy Y. Srinivasa
    • International Journal of Industrial Entomology and Biomaterials
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    • v.12 no.2
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    • pp.75-80
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    • 2006
  • Changes in the levels of protein and free amino acids in the haemolymph of three selected races of the silkworm, Bombyx mori viz., PM, $NB_4D_2$ and $CSR_2$, were investigated during 4th moult, 5th instar and pupal period. The levels of total protein in the haemolymph, increased from first day of 5th instar till sixth day. From seventh day till spinning, the protein levels decreased in all the three races. A sustained decrease in the haemolymph proteins was observed during the pupal development in all the three races. The levels of free amino acids, which were high during 4th moult, declined through the 5th age of larval development till spinning. PM showed a relatively higher free amino acid level (3.192 mg/ml) in haemolymph followed by $NB_4D_2$ (2.601 mg/ml) and $CSR_2$ (2.35 mg/ml). The free amino acid levels decreased gradually from prepupal stage but increased again at the end of pupal period. Racial differences in the changes in the levels of protein and free amino acids in the haemolymph were observed in the larvae and pupae when subjected to two high temperature regimes of $30^{\circ}C$ and $35^{\circ}C$. The results showed that high temperature induces specific changes in the metabolism (reversible thermal stress) that have different adaptive value in different races of the silkworm. Relatively higher increase in the free amino acid levels in the haemolymph of Pure Mysore presumably provides protective cover to tissues against high temperature by an increase in osmolarity and reduction in evaporative water loss. The absence of such a mechanism may be responsible for temperature susceptibility of the bivoltine races like $NB_4D_2$ and $CSR_2$.

Seasonal Occurrence and Chemial Control Effects of Eriococcus largerstroemiae Kuwana on Persimmon Trees (감나무의 주머니깍지벌레에 대한 발생생태 및 화학적 방제효과)

  • 권태영;박소득;박선도;최부술;권용정
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.295-299
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    • 1995
  • This study was conducted to find the control methods, status of overwintering and seasonal occurrence of Erioccoccus largerstroemiae on persimmon trees in Ch'ondo area, Kyongbuk province in Korea. Usually, overwintering eggs were sheltered in bark, among them the rate of alive eggs was 28.7%. The activiation of garpe-myrtle scale showed from late April, and they have three generations per year. The first occurrence of larval stage of Erioccoccus largerstroemiae starts from late June to early July, the second occurrence begins from middle August to late August, and from late June to early July, the second occurrence begins from middle August to late August, and from middle September to late September is the third, thus, three peaks of occurrence revealed in early July middle August, and late September respectively. Average number of eggs conceived in female adult was 229.3. Spray effect of chemical control showed as follows; lime sulfur with tow applications of pesticides (late June, late August) at 97.8%, machine oil with tow applications of pesticides (late June, late August) at 96.8%. And during the growing period, the spray results using three applications of pesticide only (late June, late August, amid September) showed 77.2% in field condition.

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A new method for concentration of proteins in the calcareous corpuscles separated from the spargana of Spirometra erinacei

  • PARK Yun-Kyu;PARK Jae-Hwan;GUK Sang-Mee;SHIN Eun-Hee;CHAI Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.43 no.3 s.135
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    • pp.119-122
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    • 2005
  • Calcareous corpuscles are a characteristic structure found in larval and adult stage cestddes, These corpuscles are known to contain several protein components and to possess protein-binding activity. However, the proteins bound to calcareous corpuscles in situ have not been studied. The present study was undertaken to identify the proteins on calcareous corpuscles. Calcareous corpuscles were purified from the plerocercoids (= spargana) of Spirometra erinacei, and serially dissolved using 0.1 M sulfamic acid solution. Collected supernatants were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. The results showed that only the fraction remaining after the 19th dissolved fraction contained proteins. A total of 20 protein molecules were detected in gel, with major bands at 56, 53, 46, 40, 35, 29, 28, 24.5, 21, 19, 16, 13, 10 and 8 kDa. In particular, the proteins corresponding to the 21 and 16 kDa bands were most abundant. Our results demonstrated for the first time the protein contents of the calcareous corpuscles of spargana. Further studies on the functions of these proteins are required.

ON THE EFFECTS CHLORINITIES UPON GROWTH OF EARLIER LARVAE AND POST-LARVA OF A FRESH WATER PRAWN, MACROBRACHIUM ROSENBERGI(DE MAN) (담수산새우 Macrobrachium rosenbergi (de Man)의 초기유생 및 Post-larva.의 성장에 미치는 염분량에 관하여)

  • KWON Chin Soo;UNO Yutaka;OGASAWARA Yohismitsu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.2
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    • pp.97-114
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    • 1977
  • The fresh water prawn, Macrobrachium rosenbergi(de Man) is a very common species in Indopacific region, which inhaits both fresh and brackish water in low land area, most of rivers and especially aboundant in the lower reaches which are influenced by sea water. It is one of the largest and commercial species of genus Macrobrachium, which is commonly larger than $18\~21cm$ in body length, from the basis of eye-stalked to the distal of telson. As a part of the researches in order to investigate the possibilities on transplantation and propagation of this species, this work dealt with the problems on the effects of chlorinities upon zoeal larvae and post-larvae 1). metamorphosis rate and optimum chlorinity for metamorphosis to post-larve, 2). tolerance and comparative survival rate on various chlorinties, from fresh water to sea water $(19.38\%_{\circ}\;Cl)$, which reared for six days upon each stage of zoeal larvae, 3). accomodation rate on chlonities which reared for twelve days after transmigration into variant chlorinities of the range from $3.68\%_{\circ}$ Cl to $1.53\%_{\circ}$ Cl in the way of rearing of the range from $3.82\%_{\circ}$ Cl to $11.05\%_{\circ}$ upon each stage of zoea, 4). tolerance on both of fresh and sea water upon zoeal larva and post-larva under the condition of $28^{\circ}C{\pm}1$ in temperature and feeding on Artenia salina nauplii, 5). relationship between various chlorinities and grwth of post-larvae under the condition of $28^{\circ}C$ in tmperature and feeding on meat of clam. Thus these investigations were performed in order to grope for a comfortable method on seedmass production. Up to the present, the study on the effects of chlorinity upon earlier zoeal larvae and post-larvae of Macrobrachium species has been scarcely performed by workers with the exception of Lewis(1961) and Ling (1962,, 1967), even so their works were not so detailed. On the other hand, larvae of several species of this genus were reared at the water which mixed sea water so as to carry out complete metamorphosis to post-larva by workers in order to investigate on earlier 1 arval and earlier post-larval development, such as Macrobrachium lamerrei (Rajyalakshmi, 1961), M. rosenbergi and M. nipponense (Uno and Kwoa, 1969; Kwon and Uno, 1969), M. acanthurs (Choudhury, 1970; Dobkin, 1971), M. carcinus(Choudhury, 1970), M. formosense(Shokita, 1970), M. olfersii (Duggei et al., 1975), M. novaehallandiae (Greenwood et al., 1976), M. japonicum (Kwon, 1974) and M. lar (Shokita, personal communication), and there fore it is regarded that chlorinity is, generally, one of absolute factors to rear zoeal larvae of brackish species of Macrobrachium genus. Synthetic results on this work is summarized as the follwings: 1) Zoeal larvae required different chlorinities to grow according to each stage, and generally, it is regarded that optimum range of living and growing is from $7.63\%_{\circ}Cl\to\;7.63\%_{\circ}Cl$, and while differences of metamorphsis rate, from first zoea to post-larva, is rarely found in this range, and however it occurs apparently in both of situation at $7.63\%_{\circ}Cl$ below and $16.63\%_{\circ}Cl$ above and moreover, metamorphosis rate is delayed somewhat in case of lower chlorinity as compared with high chlorinity in these situations. 2) Accomodation in each chlorinity on the range, from fresh water to sea water, is different according to larval stages and while the best of it is, generally, on the range from $14.24\%_{\circ}Cl$ to $8.28\%_{\circ}Cl$ and favorite chlorinity of zoea have a tendency to remove from high chlorinity to lower chlorinity in order to advance larval age throughout all zoeal stages, setting a conversional stage for eighta zoea stage. 3) Optimum chlorinity of living and growth upon postlarvae is on the range of $4.25\%_{\circ}Cl$ below, and in proportion as approach to fresh water, growth rate is increased. 4) Post-large are able to live better in fresh water in comparison with zoeal larvae, which are only able to live within fifteen hours, and by contraries, post-larvae are merely able to live for one day as compared with ?미 larvar, which are able to live for six days more in sea water $19.38\%_{\circ}Cl\;above$. 5) Also, in case of transmigration into higher and lower chlorinities in the way of rearing in the initial chlorinities $ 3.82\%_{\circ}Cl,\;7.14%_{\circ}Cl\;and\;11.05%_{\circ}Cl$, accoodation rate is a follow: accomodation capacity in ease of removing into higher chlorinities from lower chlorinities is increased in proportion as earlier stages, setting a conversional stage for eighth zoea stage, and by contraries, in case of advanced stages from eighth zoea it is incraesed in proportion as approach to post-larva stage in the case of transmigration into lower chlorinity from higher chlorinity. On the other hand, it is interesting that in case of reciprocal transmigration between two different chlorinitiess, each survival rate is different, and in this case, also, its accomodation in each zoea stage has a tendency to vary according to larval stages as described above, setting a conversional stage for eighth zoea stage. 6) It is likely that expension of radish pigments on body surface is directly proportional to chlorinity during the period of zoea rearing, and therefore it seems like all body surfacts of zoea larvae be radish coloured in case of higher chlorinity. 7) By the differences that each zoeal larvae, postlarvae, juvaniles and adult prawn are required different chlorinity for inhabiting in each, it is regarded that this species migrats from up steam to near the estuary of the river which the prawns inhabits commonly in natural field for spawning and growth migration. 8) It had better maintainning chlorinities according to zoeal stage for a comfortable method on seed-mass production that earlier larva stages than eighth zoea are maintained on the range from $8\%_{\circ}Cl\;to\;12\%_{\circ}Cl$ to rear, and later larva stages than eighth zoea, by contraries, are gradually regula ted-to love chlorininity of the range from $7\%_{\circ}Cl\;to\;4\%_{\circ}Cl$ according to advance for post-larva stage.

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Life Cycle of Tipula latemarginata Alexander (Diptera: Tipulidae) in Korea (한국산 애아이노각다귀, Tipula latemarginata Alexander, (파리목, 각다귀과)의 생활사)

  • Kim Dong Sang;Lee Jong Eun
    • Korean journal of applied entomology
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    • v.44 no.2
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    • pp.109-114
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    • 2005
  • This study was conducted to investigate the life cycle of Tipula latemarginata Alexander in Korea. The field surveys for the life cycle of the species were carried out at the two sites of Neaseong Stream area in Bongwha County from January 2001 to December 2002. Also many individuals of the species were reared in laboratory to investigate the life cycle. T latemarginata appeared to have three generations a year under the rearing conditions at room temperature. All the processes of life cycle of the species, when reared at room temperature from September 2001 to March 2002, were as follows: Eggs usually hatched between 4 and 10 days after oviposition. First instar larvae grew rapidly and molted to the second instar in 7-9 days. Second instar larvae spent 5-7 days for next molting and third instar period lasted approximately 8-12 days. Fourth instar larvae spent 6 weeks to 5 months for pupation. Especially the duration of fourth instar larval existence was greatly lengthened and served as the overwintering stage. The duration of pupal stage was 6-10 days. In the field, T. latemarginata also appeared to have three generations a year at the favorable habitats.

Biology of Three Species of the Genus Tipula (Diptera: Tipulidae) in Korea (한국산 Tipula속(파리목 : 각다귀과) 3종의 생태)

  • Kim, Dong-Sang;Lee, Jong-Eun
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.201-212
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    • 2007
  • This study was conducted to investigate the biology of three species, Tipula latemarginata Alexander, T. nova Walker and T. aino Alexander, of the Genus Tipula in Korea. Field surveys for the biology of the species were carried out at the three sites of Neaseong Stream area in Bongwha County from January 2001 to December 2005. Also many individuals of the species were reared in laboratory to investigate the biology. The larval habitats of the species were streams, rivers, paddy fields, damp earth or leaky revetments. The Tipula larvae were herbivorous. Newly-hatched first instar larvae fed on soft algae, and the larvae gradually fed on leaves, stems and roots of a variety of plants or hard leaf litters, as they grew. During the molting of the larvae their body kept moving from left to right and up and down. Molting usually took not more than 2 hours, and about four hours after molting, the larvae started to eat heavily. The Tipula larvae in aquatic places moved to drier land for pupation and went through short prepupal stage lasting 1-2 days for pupation at fourth instar larval stage. When emerging, the Tipula pupae placed their head and thorax on the earth, but the other parts in the earth. Emergence from the pupal case required about 20 min. to one hour. Mating of Tipula adults took place within 5-7min. after emergence and the duration of mating was about 40 min. The female adults of the species laid eggs by walking with 3 pairs of legs over the damp earth or algal beds. Their body was positioned vertically on the ground with their wings spread $120^{\circ}$ and legs landed on the surface. The oviposition usually took place from one day to 4 days after emergence and the number of eggs carried by female adults were an average of 501-760 per individual. Tipula adults didn't normally feed, and drank water only occasionally. For a few days after emergence, the adults reared in the laboratory rarely drank water. As they neared to death, however, they frequently drank water. The longevity of adults reared in the laboratory with only water during the summer was ca. 4-9 days and males usually survived a little longer than females. The longevity of T. nova was increased 3 times or more as much by feeding them 3% sugar water. Male adults of T. latemarginata outnumbered female adults by 2.6 : 1 in the fields.