Microbial dehalogenation of chlorinated compounds in anaerobic environments is well known, but the degradation of fluorinated compounds under similar conditions has rarely been described. Here, we report on the isolation of a bovine rumen bacterium that metabolizes fluoroacetate under anaerobic conditions, the mode of degradation and its presence in gut ecosystems. The bacterium was identified using 16S rRNA gene sequence analysis as belonging to the phylum Synergistetes and was designated strain MFA1. Growth was stimulated by amino acids with greater quantities of amino acids metabolized in the presence of fluoroacetate, but sugars were not fermented. Acetate, formate, propionate, isobutryate, isovalerate, ornithine and H2 were end products of amino acid metabolism. Acetate was the primary end product of fluoroacetate dehalogenation, and the amount produced correlated with the stoichiometric release of fluoride which was confirmed using fluorine nuclear magnetic resonance (19F NMR) spectroscopy. Hydrogen and formate produced in situ were consumed during dehalogenation. The growth characteristics of strain MFA1 indicated that the bacterium may gain energy via reductive dehalogenation. This is the first study to identify a bacterium that can anaerobically dehalogenate fluoroacetate. Nested 16S rRNA gene‐specific PCR assays detected the bacterium at low numbers in the gut of several herbivore species.
A study of eight commercial cattle herds grazing leucaena (Leucaena leucocephala subsp. glabrata) pastures was undertaken to determine (1) the efficacy of in vitro Synergistes jonesii inoculum (produced in an anaerobic fermenter) in degrading the dihydroxypyridone (DHP) isomers produced during digestion of leucaena forage; and (2) the persistence of the inoculum in the rumen of cattle following a period grazing non-leucaena pastures. Cattle were introduced to the leucaena pastures for an initial period varying from 17 to 71 days. Fourteen to fifteen animals were then sampled for (1) urine and blood plasma to determine toxicity status as indicated by concentration of DHP; (2) faeces for estimation of diet composition; and (3) rumen fluid for detection of S. jonesii by nested polymerase PCR analysis. After a further 42–56 days, animals were resampled as before to confirm toxicity status and inoculated with the in vitro S. jonesii inoculum; the herds were then sampled a third time (42–60 days after inoculation) to test the effectiveness of the inoculum in degrading DHP. Five of the herds were then removed from leucaena pastures for periods ranging from 80 to 120 days and returned to leucaena pastures for 21 days to check persistence of the inoculum as indicated by retention of capacity to degrade DHP. The data indicated (1) a very slow build up of capacity to degrade DHP isomers on some properties before inoculation; (2) frequent occurrence of high levels of 2,3-DHP in urine indicating partial toxin degradation, both before and after inoculation; (3) a low incidence of detection of S. jonesii in rumen fluid after inoculation based on nested PCR analysis; (4) failure of inoculation to degrade DHP on one of two properties tested; and (5) loss of capacity to degrade DHP on some properties after <4 months on alternative non-leucaena pastures. It was concluded that while most herds showed some capability to degrade DHP due to some residual capability from previous exposure, they did not achieve the same rapid and complete DHP degradation reported in the 1980s. Nevertheless, it was concluded that the in vitro inoculum was at least partially effective and should continue to be used by graziers until improved sources of inoculum and/or inoculation methodologies are demonstrated.
Fluoroacetate producing plants grow worldwide and it is believed they produce this toxic compound as a defence mechanism against grazing by herbivores. Ingestion by livestock often results in fatal poisonings, which causes significant economic problems to commercial farmers in many countries such as Australia, Brazil and South Africa. Several approaches have been adopted to protect livestock from the toxicity with limited success including fencing, toxic plant eradication and agents that bind the toxin. Genetically modified bacteria capable of degrading fluoroacetate have been able to protect ruminants from fluoroacetate toxicity under experimental conditions but concerns over the release of these microbes into the environment have prevented the application of this technology. Recently, a native bacterium from an Australian bovine rumen was isolated which can degrade fluoroacetate. This bacterium, strain MFA1, which belongs to the Synergistetes phylum degrades fluoroacetate to fluoride ions and acetate. The discovery and isolation of this bacterium provides a new opportunity to detoxify fluoroacetate in the rumen. This review focuses on fluoroacetate toxicity in ruminant livestock, the mechanism of fluoroacetate toxicity, tolerance of some animals to fluoroaceate, previous attempts to mitigate toxicity, aerobic and anaerobic microbial degradation of fluoroacetate, and future directions to overcome fluoroacetate toxicity.
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