Hydrogen utilising bacteria from the forestomach of eastern grey (Macropus giganteus) and red (Macropus rufus) kangaroos
D. Ouwerkerk A , A. J. Maguire A , L. McMillen A and A. V. Klieve A BA Animal Science, Department of Primary Industries and Fisheries, Yeerongpilly, Qld 4105, Australia.
B Corresponding author. Email: athol.klieve@dpi.qld.gov.au
Animal Production Science 49(11) 1043-1051 https://doi.org/10.1071/EA08294
Submitted: 4 December 2008 Accepted: 26 June 2009 Published: 14 October 2009
Abstract
Reductive acetogenesis is an alternative to methanogenesis for removing hydrogen produced during enteric fermentation. In Australia, kangaroos have evolved an enlarged forestomach analogous to the rumen of sheep and cattle. However, unlike sheep and cattle, kangaroos produce very little methane from enteric fermentation. From samples of gut contents from five eastern grey and three red kangaroos, we were not able to detect methanogens using a PCR protocol, but did detect the formyltetrahydrofolate synthetase (FTHFS) gene (likely to be used for reductive acetogenesis) in all animals. Isolations to recover acetogens resulted in two different classes of hydrogen consuming bacteria being isolated. The first class consisted of acetogens that possessed the FTHFS gene, which except for Clostridium glycolicum, were not closely related to any previously cultured bacteria. The second class were not acetogens but consisted of enterobacteria (Escherichia coli and Shigella) that did not possess FTHFS genes but did utilise hydrogen and produce acetate. Enumeration of the acetogens containing the FTHFS gene by real-time PCR indicated that bacteria of the taxa designated YE257 were common to all the kangaroos whereas YE266/YE273 were only detected in eastern grey kangaroos. When present, both species occurred at densities above ×106 cell equivalents per mL. C. glycolicum was not detected in the kangaroos and, unlike YE257 and YE266/273, is unlikely to play a major role in reductive acetogenesis in the foregut of kangaroos.
Acknowledgements
We thank Barbara Williams, Jessica Morgan and Ian Brock at the Animal Research Institute, DPI&F (Qld), the staff of Croxdale Research station (DPI&F) and the staff of the DNA Sequence Analysis Facility of the Griffith University for technical assistance. This work was funded by Queensland DPI&F’s seed funding initiative and Meat and Livestock Australia.
Altschul SF,
Gish W,
Miller W,
Myers EW, Lipman DJ
(1990) Basic local alignment search tool. Journal of Molecular Biology 215, 403–410.
|
CAS |
PubMed |
Barns SM,
Fundyga RE, Jeffries MW
(1994) Remarkable archaeal diversity detected in a Yellowstone-national park hot spring environment. Proceedings of the National Academy of Sciences of the United States of America 91, 1609–1613.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Breznak JA, Switzer JM
(1986) Acetate synthesis from H2 plus CO2 by termite gut microbes. Applied and Environmental Microbiology 52, 623–630.
|
CAS |
PubMed |
Burow LC,
Gobius KS,
Vanselow BA, Klieve AV
(2005) A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing E. coli (STEC). Letters in Applied Microbiology 40, 117–122.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Dellow DW,
Hume ID,
Clarke RTJ, Bauchop T
(1988) Microbial activity in the forestomach of free-living macropodid marsupials: comparisons with laboratory studies. Australian Journal of Zoology 36, 383–395.
| Crossref | GoogleScholarGoogle Scholar |
Evans PN,
Hinds LA,
Sly LI,
McSweeney CS,
Morrison M, Wright ADG
(2009) Community composition and density of methanogens in the foregut of the tammar wallaby (Macropus eugenii). Applied and Environmental Microbiology 75, 2598–2602.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Fievez V,
Mbanzamihigo L,
Piattoni F, Demeyer D
(2001) Evidence for reductive acetogenesis and its nutritional significance in ostrich hindgut as estimated from in vitro incubations. Journal of Animal Physiology and Animal Nutrition 85, 271–280.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Fonty G,
Joblin K,
Chavarot M,
Roux R,
Naylor G, Michallon F
(2007) Establishment and development of ruminal hydrogenotrophs in methanogen-free lambs. Applied and Environmental Microbiology 73, 6391–6403.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Joblin KN
(1999) Ruminal acetogens and their potential to lower ruminant methane emissions. Australian Journal of Agricultural Research 50, 1307–1313.
| Crossref | GoogleScholarGoogle Scholar |
Joblin KN,
Naylor GE, Williams AG
(1990) Effects of Methanobrevibacter smithii on xylanolytic activity of anaerobic ruminal fungi. Applied and Environmental Microbiology 56, 2287–2295.
|
CAS |
PubMed |
Karch H, Meyer T
(1989) Single primer pair for amplifying segments of distinct Shiga-like-toxin genes by polymerase chain-reaction. Journal of Clinical Microbiology 27, 2751–2757.
|
CAS |
PubMed |
Kempton TJ,
Murray RM, Leng RA
(1976) Methane production and digestibility measurements in grey kangaroo and sheep. Australian Journal of Biological Sciences 29, 209–214.
|
CAS |
PubMed |
Klieve AV,
Holroyd RG,
Turner AF, Lindsay JA
(1998) Rumen bacterial and protozoal populations in cattle being relocated in tropical Queensland. Australian Journal of Agricultural Research 49, 1153–1159.
| Crossref | GoogleScholarGoogle Scholar |
Klieve AV,
Hennessey D,
Ouwerkerk D,
Forster RJ,
Mackie RI, Attwood GT
(2003) Establishing populations of Megasphaera elsdenii YE 34 and Butyrivibrio fibrisolvens YE 44 in the rumen of cattle fed high grain diets. Journal of Applied Microbiology 95, 621–630.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Klieve AV,
O’Leary MN,
McMillen L, Ouwerkerk D
(2007) Ruminococcus bromii, identification and isolation as a dominant community member in the rumen of cattle fed a barley diet. Journal of Applied Microbiology 103, 2065–2073.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Leaphart AB, Lovell CR
(2001) Recovery and analysis of formyltetrahydrofolate synthetase gene sequences from natural populations of acetogenic bacteria. Applied and Environmental Microbiology 67, 1392–1395.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Ludwig W,
Strunk O,
Westram R,
Richter L, Meier H , et al.
(2004) ARB: a software environment for sequence data. Nucleic Acids Research 32, 1363–1371.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Maidak BL,
Cole JR,
Lilburn TG,
Parker CT, Saxman PR , et al.
(2000) The RDP (Ribosomal Database Project) continues. Nucleic Acids Research 28, 173–174.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Nollet L,
Vande Velde I, Verstraete W
(1997) Effect of the addition of Peptostreptococcus productus ATCC35244 on the gastro-intestinal microbiota and its activity, as simulated in an in vitro simulator of the human gastro-intestinal tract. Applied Microbiology and Biotechnology 48, 99–104.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Ouwerkerk D, Klieve AV
(2001) Bacterial diversity within feedlot manure. Anaerobe 7, 59–66.
| Crossref | GoogleScholarGoogle Scholar |
Ouwerkerk D,
Klieve AV, Forster RJ
(2002) Enumeration of Megasphaera elsdenii in rumen contents by real-time Taq nuclease assay. Journal of Applied Microbiology 92, 753–758.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Ouwerkerk D,
Klieve AV,
Forster RJ,
Templeton JM, Maguire AJ
(2005) Characterization of culturable anaerobic bacteria from the forestomach of an eastern grey kangaroo, Macropus giganteus. Letters in Applied Microbiology 41, 327–333.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Ovreås L,
Forney L,
Daae FL, Torsvik V
(1997) Distribution of bacterioplankton in meromictic Lake Sælenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for the 16S rRNA. Applied and Environmental Microbiology 63, 3367–3373.
| PubMed |
Posada D, Crandall KA
(1998) Modeltest: testing the model of DNA substitution. Bioinformatics (Oxford, England) 14, 817–818.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Sar C,
Mwenya B,
Santoso B,
Takaura K,
Morikawa R,
Isogai N,
Asakura Y,
Toride Y, Takahashi J
(2005) Effect of Escherichia coli W3110 on ruminal methanogenesis and nitrate/nitrite reduction in vitro. Animal Feed Science and Technology 118, 295–306.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Stackebrandt E, Charfreitag O
(1990) Partial 16S rRNA primary structure of five actinomyces species: phylogenetic implications and development of an actinomyces israelii-specific oligonucleotide probe. Journal of General Microbiology 136, 27–43.
Teather RM
(1982) Maintenance of laboratory strains of obligately anaerobic rumen bacteria. Applied and Environmental Microbiology 44, 499–501.
|
CAS |
PubMed |
von Engelhardt W,
Wolter S,
Lawrenz H, Hemsley JA
(1978) Production of methane in two non-ruminant herbivores. Comparative Biochemistry and Physiology. A. Comparative Physiology 60, 309–311.
| Crossref | GoogleScholarGoogle Scholar |
Wetzel AN, LeJeune JT
(2007) Isolation of Eschericia coli O157:H7 strains that do not produce Shiga toxin from bovine, avian and environmental sources. Letters in Applied Microbiology 45, 504–507.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |