Regeneration of protoplasts of Entomophthora egressa, a fungal pathogen of the eastern hemlock looper

1977 ◽  
Vol 55 (1) ◽  
pp. 107-113 ◽  
Author(s):  
Gary B. Dunphy ◽  
Richard A. Nolan
Keyword(s):  
Osmotic Shock ◽  
Calf Serum ◽  
Egg Yolk ◽  
Eastern Hemlock ◽  
Liquid Media ◽  
Developmental Patterns ◽  
Solid Substrates ◽  
Insect Tissue Culture ◽  
The Individual ◽  
Hemlock Looper

The regeneration patterns for protoplasts of Entomophthora egressa MacLeod and Tyrrell grown on modified Grace's insect tissue culture medium supplemented with fetal calf serum (a highly modified version of Grace's medium lacking serum and more closely approximating the hemolymph of the eastern hemlock looper), Müller-Kögler's coagulated egg yolk medium supplemented with Tristearin, and water agar are documented. Protoplasts on liquid and egg yolk media had varying degrees of common developmental patterns. On liquid media, development involving prophyphal spheres and fusion spheres predominated. Hyphae emerged from the prohyphal spheres; however, the fusion spheres did not develop further. On the coagulated egg yolk medium, the individual, non-chain, cells coalesced and developed into pleomorphs which later developed into osmotic shock-resistant postprotoplasts. A postprotoplast gave rise to a hypha which produced a terminal conidium. Later resting spores were detected. Water agar supported scant mycelial growth as compared with that on coagulated egg yolk but more than that on the liquid media. Solid substrates appeared to favor mycelial development. Regeneration could be avoided by the addition of fresh Grace's modified medium to either coagulated egg yolk plates or to cultures with Grace's modified medium. Obtaining protoplasts from hyphae and the reversion of these protoplasts to form hyphae appeared to be a fixed property of E. egressa.This report is believed to be the first for reversion of protoplasts initially produced by non-enzymic means.

NINHYDRIN-POSITIVE SUBSTANCE ANALYSIS OF LARVAL HEMOLYMPH OF THE EASTERN HEMLOCK LOOPER, LAMBDINA FISCELLARIA FISCELLARIA (LEPIDOPTERA: GEOMETRIDAE) AND GROWTH OF ENTOMOPHTHORA EGRESSA PROTOPLASTS

1977 ◽  
Vol 109 (3) ◽  
pp. 341-346 ◽  
Author(s):  
Gary B. Dunphy ◽  
Richard A. Nolan ◽  
Imre S. Otvos
Keyword(s):  
Generation Time ◽  
Analytical Study ◽  
Culture Medium ◽  
Fungal Growth ◽  
Eastern Hemlock ◽  
Larval Hemolymph ◽  
Insect Tissue Culture ◽  
High Concentrations ◽  
Hemlock Looper ◽  
Current Report

AbstractA ninhydrin-positive substance analysis of fourth instar larval hemolymph of the eastern hemlock looper, Lambdina fiscellaria fiscellaria (Guenée), was conducted. Two separate larval populations were reared on a defined diet during the early stages and later on fresh, young balsam fir foliage. The results indicated the presence of at least 38 compounds. L-glutamine and(or) L-asparagine, L-histidine, and L-lysine occurred at high concentrations (> 5000 nM/ml). Ammonia, L-arginine, L-threonine, L-serine, L-glutamic acid, glycine, L-alanine, L-valine, L-isoleucine, L-leucine, L-tyrosine, and L-phenylalanine occurred at intermediate levels (1000–5000 nM/ml). The current report of the presence of L-1-methylhistidine and L-3-methylhistidine is the second report of any methylated derivative of histidine in insect hemolymph. The results of the analytical study were used to modify the composition of Grace’s insect tissue culture medium which had previously been found to support growth of protoplasts of the looper pathogen Entomophthora egressa MacLeod and Tyrrell. Fungal growth indicated a shorter generation time (4.7 h) on the medium modified to more closely approximate the looper hemolymph as compared with the generation time (6.2 h) on Grace’s medium.

Haemoflagellates: commercially available liquid media for rapid cultivation

Parasitology ◽  
1978 ◽  
Vol 76 (3) ◽  
pp. 309-316 ◽  
Author(s):  
L. D. Hendricks ◽  
D. E. Wood ◽  
M. E. Hajduk
Keyword(s):  
Tissue Culture ◽  
Calf Serum ◽  
Laboratory Animals ◽  
Foetal Calf Serum ◽  
Liquid Media ◽  
Culture Cells ◽  
Laboratory Rodents ◽  
Freeze Dried ◽  
Insect Tissue Culture ◽  
The Media

SummaryThe successful cultivation of a variety of haemoflagellates in three different liquid media is reported. These media include medium 199, Grace's insect tissue-culture medium and Schneider's drosophila medium, each in combination with 30% (v/v) foetal calf serum. These media were used to cultivate Old and New World species of visceral and cutaneous human Leishmania, as well as Leishmania species isolated from sandflies, rodents, and reptiles. Four strains of Trypanosoma cruzi, an isolate of T. rangeli and an isolate of T. lewisi have also been cultivated in these media. One or more of these media have been used to cultivate 121 strains of haemoflagellates, including at least 14 different species (11 Leishmania and 3 Trypanosoma) and many geographic isolates or strains. The Leishmania include L. braziliensis, L. peruviana, L. mexicana, L. tropica, L. donovani, L. chagasi, L. enriettii, L. hertigi, L. hoogstraali, L. adleri, and L. agamae. Using the Schneider's based medium, we have obtained primary isolates of both cutaneous and visceral Leishmania of man and of experimentally infected laboratory rodents and canines. Freeze-dried preparations of the Schneider's based medium that were reconstituted with distilled water after 24 months of storage at ambient temperature have proven to be suitable cultivation media. This feature makes the media valuable field tools.The various species of human Leishmania cultivated in these media have in our experience demonstrated no differences in growth rate, viability after liquid nitrogen preservation, or infectivity for laboratory animals and tissue-culture cells compared with promastigotes derived from blood-agar cultivation.

Morphogenesis of protoplasts of Entomophthora egressa in simplified culture media

1977 ◽  
Vol 55 (24) ◽  
pp. 3046-3053 ◽  
Author(s):  
Gary B. Dunphy ◽  
Richard A. Nolan
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Growth Cycle ◽  
Protoplast Regeneration ◽  
Liquid Media ◽  
Insect Tissue Culture ◽  
Hyphal Bodies ◽  
The Media ◽  
The Individual ◽  
Mycelial Development

Protoplasts of Entomophthora egressa MacLeod and Tyrrell were grown in Grace's medium and in two simplified liquid media. These latter media contained reduced numbers of amino acids as compared with Grace's insect tissue culture medium. The pattern of protoplast regeneration was the same in all three media used, but additional stages in the morphogenetic sequence werefound. A spherical mesoprotoplast and an elliptical mesoprotoplast stage with actively moving filopodia were detected early in the growth cycle. The clumping of rod-shaped and spherical hyphal bodies was observed. Mycelial development was very pronounced by 89 h of incubation. The osmolarity and pH of the media were unchanged before this time. The individual hyphae were predominantly initiated by spherical hyphal bodies.All stages of regeneration produced spindle-shaped protoplasts when added to fresh media. The inoculation of spent media with spindle-shaped protoplasts favoured an accelerated morpho-genesis of the cells through the presently reported stages.

scholarly journals Dynamic Paths of Complexity and Accuracy in Second Language Speech: A Longitudinal Case Study of Chinese Learners

2019 ◽  
Vol 41 (6) ◽  
pp. 855-877 ◽  
Author(s):  
Hanjing Yu ◽  
Wander Lowie
Keyword(s):  
Oral Language ◽  
Theory Development ◽  
Longitudinal Case Study ◽  
Dynamic Interactions ◽  
Developmental Patterns ◽  
Individual Level ◽  
Complex Dynamic Systems ◽  
Chinese Learners ◽  
Oral Development ◽  
The Individual

Abstract The present study investigated the developmental patterns of Chinese EFL(AQ) learners’ oral language in terms of complexity and accuracy and looked into the dynamic interactions between them within the framework of Complex Dynamic Systems Theory (CDST). The data were analysed using dynamic analyses (moving min–max graphs, moving correlations and Monte Carlo Simulations). It was found that, firstly, at the group level, the general developmental trends of both complexity and accuracy showed improvements. Secondly, at the individual level, the developmental patterns were non-linear and dynamic with high degrees of variability, and individual language development was influenced by the initial states. Thirdly, the analyses revealed a complex interplay between complexity and accuracy, which gradually shifted from a clearly competitive relationship during the early stages to a supportive relationship in later stages. This shift in interaction shows that complexity goes hand in hand with accuracy, which corroborates the interconnectedness of subsystems as one of the major characteristics of CDST. The findings confirm the applicability of CDST approaches to L2 oral development and carry valuable implications for CDST theory development and oral language teaching.

Characterization of the Soil Hydromorphic Conditions in a Paludified Dunefield during the Mid-Holocene Hemlock Decline near Québec City, Québec

1996 ◽  
Vol 46 (3) ◽  
pp. 281-297 ◽  
Author(s):  
Najat Bhiry ◽  
Louise Filion
Keyword(s):  
Soil Fauna ◽  
Tsuga Canadensis ◽  
Eastern Hemlock ◽  
Plant Colonization ◽  
Quebec City ◽  
Hemlock Decline ◽  
Short Period ◽  
Time Required ◽  
Hemlock Looper

The mid-Holocene eastern hemlock [Tsuga canadensis L. (Carr.)] decline has been recently attributed to the activity of insect defoliators. N. Bihiry and L. Filion, Quaternary Research 45,312–320 (1996). In this study, soil hydromorphic conditions were investigated for the period 6800–3200 yr B.P. using micromorphological data from a peat section from a swale in a paludified dunefield in southern Québec. After a short period of plant colonization in shallow pools between 6800 and 6400 yr B.P., mesic conditions predominated in the interdune before the decline (6400–4900 yr B.P.), as evidenced by strong bioturbation and abundance of excrements from the soil fauna. During the decline, a shift from mesic to wet conditions occurred (4900–4100 yr B.P.), although xeric to mesic conditions persisted on dune ridges until at least 4200 yr B.P. Wetness culminated when beaver occupied the site (4100–3750 yr B.P.). Hemlock needles with chewing damage typical of hemlock looper (Lambdina fiscellaria) feeding were identified at levels dated 4900, 4600, and 4200 yr B.P., respectively, implying that the hemlock decline was associated with at least three defoliation events. The ca. 400-yr interval between these events likely represents the time required for this late-sucessional tree species to recover.

scholarly journals Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
M. S. Arias ◽  
C. F. Cazapal-Monteiro ◽  
J. Suárez ◽  
S. Miguélez ◽  
I. Francisco ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Fungal Spores ◽  
Fungal Growth ◽  
Mucor Circinelloides ◽  
Parasitic Infections ◽  
Duddingtonia Flagrans ◽  
Liquid Media ◽  
Predator Species ◽  
Third Stage ◽  
The Individual

Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs) mainly caused by roundworms (ascarids) and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides) and other predator (Duddingtonia flagrans). Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4· 12 H2O, and wheat (Triticum aestivum)). A Fasciola hepatica recombinant protein (FhrAPS) was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts ofMucorspores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production ofDuddingtoniaspores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability ofT. caniseggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor) and a predator species (Duddingtonia) for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs.

THE AEROBIC BACTERIA ASSOCIATED WITH THE EASTERN HEMLOCK LOOPER LAMBDINA FISCELLARIA (GN.)

1952 ◽  
Vol 30 (4) ◽  
pp. 208-212 ◽  
Author(s):  
T. A. Angus
Keyword(s):  
Bacterial Flora ◽  
Eastern Hemlock ◽  
Soil Types ◽  
Aerobic Bacteria ◽  
Insect Larvae ◽  
Digestive Process ◽  
Hemlock Looper

A study was made of healthy hemlock loopers (Lambdina fiscellaria Gn.) to determine whether a specific bacterial flora is associated with this insect. Larvae, pupae, adults, and eggs were examined by a number of methods and the bacteria found were cultured and classified. Organisms belonging to nine genera of bacteria were isolated and they were principally of foliage-contaminating or soil types that could be ingested with the insects' food. Few of the ingested bacteria survive the digestive process. It is concluded that the bacterial flora is adventitious.

An inexpensive medium for mass fermentation production of Entomophaga aulicae hyphal bodies competent to form conidia

1993 ◽  
Vol 39 (6) ◽  
pp. 588-593 ◽  
Author(s):  
Richard A. Nolan
Keyword(s):  
Growth Medium ◽  
Entomopathogenic Fungus ◽  
Basal Medium ◽  
Fermentation Medium ◽  
Eastern Hemlock ◽  
Developmental Sequence ◽  
Hyphal Bodies ◽  
Hyphal Body ◽  
Hemlock Looper ◽  
Single Medium

A mass fermentation medium for growth and morphogenesis of the entomopathogenic fungus Entomophaga aulicae was developed. This fungus is a major pathogen of larval eastern hemlock looper and spruce budworm. The medium consists of a basal medium plus 0.8% tryptic soy broth and 0.4% calcium caseinate. This medium is a major breakthrough in that (i) the E. aulicae developmental sequence from protoplast inoculum to hyphal bodies competent to form conidia can be carried out in a single medium without adjustment, (ii) by examining the fermentation product it can be determined if conidia can be produced prior to engaging in costly field spraying, (iii) this medium supports the growth of E. aulicae isolates from different geographical areas, (iv) the medium is relatively inexpensive, (v) the hyphal bodies are easily separated from the spent growth medium, and (vi) the hyphal body yield is high.Key words: Entomophaga aulicae, mass fermentation medium, hyphal bodies, conidia, insect biocontrol.

scholarly journals EFFECT OF EXTRACTED LOW DENSITY LIPOPROTEINS SUBSTITUTION ON SOME POST CRYOPRESERVATION SEMEN QUALITY OF HOLSTEIN BULLS

2016 ◽  
Vol 47 (1) ◽  
Author(s):  
Al-Ani & et al.
Keyword(s):  
Semen Quality ◽  
Plant Protection ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Egg Yolk ◽  
Low Density Lipoproteins ◽  
Control Group ◽  
Low Density ◽  
Sperm Plasma Membrane ◽  
The Individual

The aim of this study was to investigate the effect of replacement of low density Lipoproteins (LDL) in different concentrations  ,exctracted fram egg yolk in the Tris diluents of the Holestion Freizan bulls semen after cooling and different cryopreservation periods . This study was done during the period From September/2014 to March/2015 and a two parts periods . First from September to November /2014 to done the extraction of the low density Lipoprotein which it have be succifully  done at the viruses Laportary  which belonge to the diagnosis department /state Board of the plant protection /Ministry of Agriclturd (Abu_Gruiab/west of Baghdad).The LDL extraction was done by a procedure with different steps from fresh egg Yolk and the packeges were done after that and preservie it at Refregartor( 5c ,one week only) until used . The second period was began from December/2014 to March/2015 At the Artificial insemination Department /Abu_Gruab which belonge to the General company of the Animal Resources Department services/Ministry of Agricultural (25 Km west of Baghdad ). Four Freigan Bulls at 3_3.5 year old were used to collection the semen (1 eijaculate / bull / week) by artificial vagina . 1 millelitter of semen from each bull was taken and make pooling . Egg yolks was  added 20% to the control group it is substituted for the rest of the Low density lipoproteins and different concentrations , amounting to (8% LDL and 10% LDL and 12% LDL). And study the effect of these additions in recipes semen during different periods of Save (cooling 5c) and freeze after(48 hours and one month and two months and three months) .The results of the study reveled that the 8% LDL treatment (T1) group have a significant effect (p≤0.05) on the individual motility ,sperm plasma membrane and acrosome integrity in comparable with the control group at different  periods of the preservation.

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