Isolation of an Anaerobic Mycoplasma from Avian Cell Cultures and Some of Its Effects on Marek's Disease Virus

Simmons, Donald G.; Lukert, P. D.

doi:10.2307/1588668

Cited by 5 publications

(2 citation statements)

References 18 publications

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“…The obligately anaerobic nature of the mycoplasmas reported here distinguishes them from other mycoplasmas that have been described previously, although mycoplasmas that require reduced oxygen tension for initial isolation but lose this requirement upon repeated subculture have been described (24,27). Strictly anaerobic conditions are required for the growth of the organisms described in this study even though they have been transferred more than 50 times over a 2-year period.…”

Section: Discussionmentioning

confidence: 45%

Anaeroplasma abactoclasticum gen.nov., sp.nov.: an Obligately Anaerobic Mycoplasma from the Rumen

Robinson

Allison

Hartman

1975

International Journal of Systematic Bacteriology

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Obligately anaerobic, filterable microorganisms have been isolated from the rumens of cattle and sheep. On the basis of typical colonial appearance, lack of cell wall, filterability through a 450-nm membrane filter, absence of reversion to a bacterial form, and inhibition of growth by homologous antiserum, these organisms fit the description for organisms in the order Mycoplasmatales. They exhibit unique cultural, biochemical, and serological properties, and we have thus placed them in a new genus, Anaeroplasma. A . abactoclmticum is the type species of the new genus. Strain 6-1, a sterol-requiring strain, has been designated as the type strain of A . abactoclasticum and has been deposited in the American Type Culture Collection as ATCC 27879; a non-sterol-requiring strain, number 161, has been deposited as ATCC 27880. The relationship of Anaeroplasma abactoclasticum to Acholesplusma bactoclmticum, an anaerobic mycoplasma possessing extracellular proteolytic and bactoclastic enzymes, is discussed.In 1966, Hungate (13) described an obligately anaerobic rumen microorganism that lysed bacterial cells. Subsequently, the organism was characterized by Robinson and Hungate (21) as a mycoplasma. The mycoplasma possessed lytic enzymes that caused the partial digestion of killed gram-negative bacteria or casein.Using methods similar to those of Robinson and Hungate ( Z l ) , we also isolated bacteriolytic mycoplasmas from the rumens of cattle and sheep. In addition, we discovered that anaerobic mycoplasmas not digesting bacterial cells were present at higher concentrations than the lytic agents.This paper describes the results of experiments conducted to characterize these new mycoplasmas and contains a proposal for plating them in a new genus. MATERIALS AND METHODS Media.Primary isolation medium (PIM) was similar to medium 98-5 previously used to culture rumen bacteria (7), except that autoclaved Escherichia coli cells (0.596, wt/vol) and penicillin G (1,OOO U/ml) were added.Clarified rumen fluid broth (CRFB) medium was similar to the PIM, except that Trypticase (0.2%) and yeast extract (0.05%) were added and the percentages of glucose, cellobiose, and starch were increased (0.2% of each instead of 0.05%); agar, penicillin, and E. coli cells were deleted.Modified medium 10 (MM-10) was medium 10 of Caldwell and Bryant (lo), except that increased concentrations of glucose, cellobiose, and starch (0.2% and instead of 0.05%) were used and sodium sulfide and agar were deleted. Medium A was similar to MM-10, except that Trypticase was extracted five times by shaking a 2% (wt/vol) solution with 2 volumes of ethyl ether and yeast extract was replaced by a mixture of B vitamins (8).Medium B was similar to MM-10, except that 40% clarified rumen fluid was added and the only carbohydrates used were 6 mg of soluble starch and 2 mg of uniformly 14C-labeled starch (10.3 pCi/mg; Amersham/Searle, Inc., Arlington Heights, Ill.) per 10 ml of medium.Preparation of media and supplements. The anaerobic methods used were essentially those of H...

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Section: Discussionmentioning

confidence: 45%

Anaeroplasma abactoclasticum gen.nov., sp.nov.: an Obligately Anaerobic Mycoplasma from the Rumen

Robinson

Allison

Hartman

1975

International Journal of Systematic Bacteriology

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“…Studies of the nutritional requirements of classical aerobic mycoplasmas have been reviewed (Razin, 1969;Razin, 1973). It is generally believed that nutritionally in adequate artificial media has prevented the cultivation of other mycoplasmas in nature (Razin, 1969;Simmons and Lukert, 1972). It has also been noted that the formation of the typical mycoplasma "fried-egg" colonial morphology was dependent on the nutritional and physical properties of the medium (Razin, 1969).…”

Section: Anaeroplasma Nutrition Studiesmentioning

confidence: 99%

Anaeroplasma bactoclasticum extracellular amylase

Jost

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Flow chart of ultrafiltration to separate amylase by molecular weight 48 Comparison of Anaeroplasma bactoclasticum amylase activity by the disc-plate assay Comparison of the effect of pH on Anaero plasma bactoclasticum amylase in various assay buffers containing 1 mM CaClg Comparison of Anaeroplasma bactoclasticum amylase production in rumen fluid and nonrumen fluid based media Comparison of Anaeroplasma bactoclasticum amylase production in different basal media Growth study of Anaeroplasma bactoclasticum in amylase production medium Anaeroplasma bactoclasticum amylase pro duction throughout growth in amylase pro duction medium Amylase activity in the supernatant fluid after the stepwise addition of ammonium sulfate Anaeroplasma bactoclasticum amylase pro duction Elution pattern of Anaeroplasma bacto clasticum partially purified amylase from a Sephadex G-200 column Elution pattern of Anaeroplasma bacto clasticum partially purified amylase from a Biogel A 0.5 m column Elution pattern of Anaeroplasma bacto clasticum partially purified amylase from a DEAE-Sepharose CL-6B column 91 93 98 148 Amylase activity in the supernatant fluid after the stepwise addition of ammonium sulfate 151

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