Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics

Axenic cultures of free-living aerobic ciliates, such as Tetrahymena thermophila and Paramecium aurelia, have been established and routinely used in laboratory research, greatly facilitating, or enabling characterization of their metabolism, physiology, and ecology. Ruminal protozoa are anaerobic ci...

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Main Authors: Tansol Park, Tea Meulia, Jeffrey L. Firkins, Zhongtang Yu
Format: Article
Language:English
Published: Frontiers Media S.A. 2017-06-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fmicb.2017.01189/full
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author Tansol Park
Tea Meulia
Jeffrey L. Firkins
Zhongtang Yu
author_facet Tansol Park
Tea Meulia
Jeffrey L. Firkins
Zhongtang Yu
author_sort Tansol Park
collection DOAJ
description Axenic cultures of free-living aerobic ciliates, such as Tetrahymena thermophila and Paramecium aurelia, have been established and routinely used in laboratory research, greatly facilitating, or enabling characterization of their metabolism, physiology, and ecology. Ruminal protozoa are anaerobic ciliates, and they play important roles in feed digestion and fermentation. Although, repeatedly attempted, no laboratory-maintainable axenic culture of ruminal ciliates has been established. When axenic ciliate cultures are developed, antibiotics are required to eliminate the accompanying bacteria. Ruminal ciliates gradually lose viability upon antibiotic treatments, and the resultant axenic cultures can only last for short periods of time. The objective of this study was to evaluate eight antibiotics that have been evaluated in developing axenic cultures of ruminal ciliates, for their toxicity to Entodinium caudatum, which is the most predominant ruminal ciliate species. Scanning and transmission electron microscopy (TEM) showed that the antibiotics damaged both the cell surface and nuclei of E. caudatum and increased accumulation of intracellular glycogen. Combinations of the three least toxic antibiotics failed to eliminate the bacteria that are present in the E. caudatum culture. The combination of ampicillin, carbenicillin, streptomycin, and oxytetracycline was able to eliminate all the bacteria, but the resultant axenic E. caudatum culture gradually lost viability. Adding the bacterial fraction (live) separated from an untreated E. caudatum culture reversed the viability decline and recovered the growth of the treated E. caudatum culture, whereas feeding nine strains of live bacteria isolated from E. caudatum cells, either individually or in combination, could not. Nutritional and metabolic dependence on its associated bacteria, accompanied with direct and indirect inhibition by antibiotics, makes it difficult to establish an axenic culture of E. caudatum. Monoxenic or polyxenic cultures of E. caudatum could be developed if the essential symbiotic partner(s) can be identified.
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spelling doaj.art-b32da8aa3381461ca29efd2d5e0dc2e62022-12-22T00:55:17ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-06-01810.3389/fmicb.2017.01189254418Inhibition of the Rumen Ciliate Entodinium caudatum by AntibioticsTansol Park0Tea Meulia1Jeffrey L. Firkins2Zhongtang Yu3Department of Animal Sciences, The Ohio State UniversityColumbus, OH, United StatesMolecular and Cellular Imaging Center, Ohio Agricultural Research and Development Center and the Department of Plant Pathology, Ohio State UniversityWooster, OH, United StatesDepartment of Animal Sciences, The Ohio State UniversityColumbus, OH, United StatesDepartment of Animal Sciences, The Ohio State UniversityColumbus, OH, United StatesAxenic cultures of free-living aerobic ciliates, such as Tetrahymena thermophila and Paramecium aurelia, have been established and routinely used in laboratory research, greatly facilitating, or enabling characterization of their metabolism, physiology, and ecology. Ruminal protozoa are anaerobic ciliates, and they play important roles in feed digestion and fermentation. Although, repeatedly attempted, no laboratory-maintainable axenic culture of ruminal ciliates has been established. When axenic ciliate cultures are developed, antibiotics are required to eliminate the accompanying bacteria. Ruminal ciliates gradually lose viability upon antibiotic treatments, and the resultant axenic cultures can only last for short periods of time. The objective of this study was to evaluate eight antibiotics that have been evaluated in developing axenic cultures of ruminal ciliates, for their toxicity to Entodinium caudatum, which is the most predominant ruminal ciliate species. Scanning and transmission electron microscopy (TEM) showed that the antibiotics damaged both the cell surface and nuclei of E. caudatum and increased accumulation of intracellular glycogen. Combinations of the three least toxic antibiotics failed to eliminate the bacteria that are present in the E. caudatum culture. The combination of ampicillin, carbenicillin, streptomycin, and oxytetracycline was able to eliminate all the bacteria, but the resultant axenic E. caudatum culture gradually lost viability. Adding the bacterial fraction (live) separated from an untreated E. caudatum culture reversed the viability decline and recovered the growth of the treated E. caudatum culture, whereas feeding nine strains of live bacteria isolated from E. caudatum cells, either individually or in combination, could not. Nutritional and metabolic dependence on its associated bacteria, accompanied with direct and indirect inhibition by antibiotics, makes it difficult to establish an axenic culture of E. caudatum. Monoxenic or polyxenic cultures of E. caudatum could be developed if the essential symbiotic partner(s) can be identified.http://journal.frontiersin.org/article/10.3389/fmicb.2017.01189/fullantibioticsassociated bacteriaaxenic cultureEntodiniumruminal protozoa
spellingShingle Tansol Park
Tea Meulia
Jeffrey L. Firkins
Zhongtang Yu
Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
Frontiers in Microbiology
antibiotics
associated bacteria
axenic culture
Entodinium
ruminal protozoa
title Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
title_full Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
title_fullStr Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
title_full_unstemmed Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
title_short Inhibition of the Rumen Ciliate Entodinium caudatum by Antibiotics
title_sort inhibition of the rumen ciliate entodinium caudatum by antibiotics
topic antibiotics
associated bacteria
axenic culture
Entodinium
ruminal protozoa
url http://journal.frontiersin.org/article/10.3389/fmicb.2017.01189/full
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AT jeffreylfirkins inhibitionoftherumenciliateentodiniumcaudatumbyantibiotics
AT zhongtangyu inhibitionoftherumenciliateentodiniumcaudatumbyantibiotics