Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

The efficacy of a cultured Synergistes jonesii inoculum to control hydroxypyridone toxicity in Bos indicus steers fed leucaena/grass diets

Michael J. Halliday A D , Hayley E. Giles A , Jagadish Padmanabha B , Chris S. McSweeney B , Scott A. Dalzell C and H. Max Shelton A
+ Author Affiliations
- Author Affiliations

A School of Agriculture and Food Science, The University of Queensland, St Lucia, Qld 4072, Australia.

B CSIRO Agriculture, St Lucia, Qld 4067, Australia.

C Ex-School of Agriculture and Food Science, The University of Queensland, St Lucia, Qld 4072, Australia.

D Corresponding author. Email: m.halliday@uq.edu.au

Animal Production Science 59(4) 696-708 https://doi.org/10.1071/AN17853
Submitted: 25 October 2016  Accepted: 31 January 2018   Published: 19 April 2018

Abstract

An experiment was conducted to investigate the efficacy of a cultured Synergistes jonesii inoculum in degrading the Leucaena leucocephala (leucaena) toxins: 3-hydroxy-4(1H)-pyridone and 3-hydroxy-2(1H)-pyridone (3,4- and 2,3-DHP). Sixteen stall-housed Bos indicus steers naïve to leucaena were fed varying combinations of forage-harvested leucaena and Chloris gayana (Rhodes grass). Dietary treatments, offered at 25 g dry matter/kg LW.day, were: 25% leucaena; 50% leucaena; 100% leucaena; and 50% leucaena, switched to 50% Medicago sativa (lucerne) after 6 weeks at time of inoculation. The experiment was 10 weeks in duration, consisting of a 6-week pre-inoculation period, followed by inoculation with cultured S. jonesii, and a 4-week post-inoculation period. Mean daily dry matter intake was recorded. Twenty-four-hour urine collections and rumen fluid samples were obtained weekly for estimation of total urinary DHP, and detection of S. jonesii using nested polymerase chain reaction analysis including presence of single nucleotide polymorphisms (SNP), respectively. In the pre-inoculation period, total urinary DHP increased quickly to high levels, then gradually declined after Week 3 with 2,3-DHP the dominant isomer through to Week 6. Indigenous strains of S. jonesii were sporadically detected by PCR analysis, indicating S. jonesii was present before inoculation but at the lower limits of detection. After inoculation there was no change in the rate of total DHP degradation or the frequency of detection of S. jonesii, although there was increased rate of degradation of 2,3-DHP. SNP indicated the presence of different strains of S. jonesii in both indigenous and cultured S. jonesii. DMI was low, especially in the 100% treatment, due in part to the high stem content of the forage-harvested leucaena and probable DHP toxicosis. It was concluded that the cultured S. jonesii inoculum did not fully protect animals against leucaena toxicity.

Additional keywords: 16S PCR, dihydroxypyridine, mimosine, rumen fluid, SNP.


References

Allison MJ, Hammond AC, Jones RJ (1990) Detection of ruminal bacteria that degrade toxic dihydroxypyridine compounds produced from mimosine. Applied and Environmental Microbiology 56, 590–594.

Allison MJ, Mayberry WR, McSweeney CS, Stahl DA (1992) Synergistes jonesii, gen. nov., sp. nov.: a rumen bacterium that degrades toxic pyridinediols. Systematic and Applied Microbiology 15, 522–529.
Synergistes jonesii, gen. nov., sp. nov.: a rumen bacterium that degrades toxic pyridinediols.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXisFWhsrY%3D&md5=20429a5d902524f87ebbced1016ba469CAS |

Andrew WM, Williams MJ, Allison MJ, Chase CC, Chambliss CG, Kalmbacher RS, Kunkle WE (2000) Detection of Synergistes jonesii in cattle and sheep feces. In ‘Proceedings of the 37th North American alfalfa improvement conference’. (Ed. M Phillips) pp. 165–168. (American Forage & Grassland Council: Madison, WI)

Aung A, ter Meulen U, Gessler F, Böhnel H (2011) Isolation of mimosine degrading bacteria from rumen juice and mass production by Göttingen bioreactor technology. Journal of Agricultural Science and Technology A 1, 764–772.

Christie GS, Lee CP, Hegarty MP (1979) Antithyroid properties of 3-hydroxy-4(1H)-pyridone: antiperoxidase activity and effect on thyroid function. Endocrinology 105, 342–347.

D’Mello JPF (1992) Chemical constraints to the use of tropical legumes in animal nutrition. Animal Feed Science and Technology 38, 237–261.
Chemical constraints to the use of tropical legumes in animal nutrition.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38XmsFSjsbg%3D&md5=503b8a7df0b189687fbf1606fa8da455CAS |

Dalzell SA, Burnett DJ, Dowsett JE, Forbes VE, Shelton HM (2012) Prevalence of mimosine and DHP toxicity in cattle grazing Leucaena leucocephala pastures in Queensland, Australia. Animal Production Science 52, 365–372.
Prevalence of mimosine and DHP toxicity in cattle grazing Leucaena leucocephala pastures in Queensland, Australia.Crossref | GoogleScholarGoogle Scholar |

Dixon RM, Coates DB (2008) Diet quality and liveweight gain of steers grazing Leucaena-grass pasture estimated with faecal near infrared reflectance spectroscopy (F.NIRS). Australian Journal of Experimental Agriculture 48, 835–842.
Diet quality and liveweight gain of steers grazing Leucaena-grass pasture estimated with faecal near infrared reflectance spectroscopy (F.NIRS).Crossref | GoogleScholarGoogle Scholar |

Domínguez-Bello MG, Lovera M, Rincón MT (1997) Characteristics of dihydroxypyridine-degrading activity in the rumen bacterium Synergistes jonesii. FEMS Microbiology Ecology 23, 361–365.
Characteristics of dihydroxypyridine-degrading activity in the rumen bacterium Synergistes jonesii.Crossref | GoogleScholarGoogle Scholar |

Elliott R, Norton BW, Milton JTB, Ford CW (1985) Effects of molasses on mimosine metabolism in goats fed fresh and dried leucaena with barley straw. Australian Journal of Agricultural Research 36, 867–875.
Effects of molasses on mimosine metabolism in goats fed fresh and dried leucaena with barley straw.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL28XkvF2lsQ%3D%3D&md5=d673665e0aad66553d96e1108e32941eCAS |

Gagen EJ, Denman SE, Padmanabha J, Zadbuke S, Al Jassim R, Morrison M, McSweeney CS (2010) Functional gene analysis suggests different acetogen populations in the bovine rumen and Tammar wallaby forestomach. Applied and Environmental Microbiology 76, 7785–7795.
Functional gene analysis suggests different acetogen populations in the bovine rumen and Tammar wallaby forestomach.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXktFKnsQ%3D%3D&md5=f7471675734e5d1fab51986d7bb6d5b2CAS |

Ghosh MK, Samiran B (2007) Mimosine toxicity – a problem of Leucaena feeding in ruminants. Asian Journal of Animal and Veterinary Advances 2, 63–73.
Mimosine toxicity – a problem of Leucaena feeding in ruminants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtVersb3E&md5=55019a7d94a83ba2e6416534af588ee8CAS |

Ghosh MK, Atreja PP, Buragohain R, Bandyopadhyay S (2007) Influence of short-term Leucaena leucocephala feeding on milk yield and its composition, thyroid hormones, enzyme activity, and secretion of mimosine and its metabolites in milk of cattle. The Journal of Agricultural Science 145, 407–414.
Influence of short-term Leucaena leucocephala feeding on milk yield and its composition, thyroid hormones, enzyme activity, and secretion of mimosine and its metabolites in milk of cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXptVSgu7o%3D&md5=f9038dfaf6cb1f5127b3b54872135c6bCAS |

Graham SR, Dalzell SA, Nguyen TN, Davis CK, Greenway D, McSweeney CS, Shelton HM (2013) Efficacy, persistence and presence of Synergistes jonesii inoculum in cattle grazing leucaena in Queensland: On-farm observations pre- and post-inoculation. Animal Production Science 53, 1065–1074.
Efficacy, persistence and presence of Synergistes jonesii inoculum in cattle grazing leucaena in Queensland: On-farm observations pre- and post-inoculation.Crossref | GoogleScholarGoogle Scholar |

Gupta HK, Atreja (1999) Influence of feeding increasing levels of leucaena leaf meal on the performance of milch goats and metabolism of mimosine and 3-hydroxy-4(1H)-pyridone. Animal Feed Science and Technology 78, 159–167.
Influence of feeding increasing levels of leucaena leaf meal on the performance of milch goats and metabolism of mimosine and 3-hydroxy-4(1H)-pyridone.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXhsVWkt74%3D&md5=ef79b7b07ccfc6a5942fd93797e5852bCAS |

Halliday MJ, Giles HE, Shelton HM (2014) The incidence of high levels of urinary 2,3-DHP in ruminants consuming Leucaena leucocephala without clinical signs of toxicity. In ‘Proceedings of the 30th biennial conference of the Australian Society of Animal Production’. (Eds S Hatcher, GL Krebs, BWB Holman) p. 180. (The Australian Society of Animal Production: Canberra)

Hammond AC, Allison MJ, Williams MJ, Prine GM, Bates DB (1989) Prevention of leucaena toxicosis of cattle in Florida by ruminal inoculation with 3-hydroxy-4-(1H)-pyridone-degrading bacteria. American Journal of Veterinary Research 50, 2176–2180.

Hegarty MP, Lee CP, Christie GS, Court RD, Haydock KP (1979) The goiterogen 3-hydroxy-4(1H)-pyridone, a ruminal metabolite from Leucaena leucocephala – effects in mice and rats. Australian Journal of Biological Sciences 32, 27–40.

Jetana T, Thongruay S, Uswang S, Hengtrakulsin R (2012) A comparative study on mimosine, 3,4-dihydroxy pyridone (3,4-DHP) and 2,3-dihydroxy pyridone (2,3-DHP), purine derivatives (PD) excretion in the urine, thyroid hormone and blood metabolites profiles of Thai swamp buffalo (Bubalus bubalis) and Murrah buffalo (Bubalus bubalis). Tropical Animal Health and Production 44, 887–897.
A comparative study on mimosine, 3,4-dihydroxy pyridone (3,4-DHP) and 2,3-dihydroxy pyridone (2,3-DHP), purine derivatives (PD) excretion in the urine, thyroid hormone and blood metabolites profiles of Thai swamp buffalo (Bubalus bubalis) and Murrah buffalo (Bubalus bubalis).Crossref | GoogleScholarGoogle Scholar |

Jones RJ (1979) The value of Leucaena leucocephala as a feed for ruminants in the tropics. World Animal Review 31, 13–23.

Jones RJ (1981) Does ruminal metabolism of mimosine explain the absence of Leucaena toxicity in Hawaii? Australian Veterinary Journal 57, 55–56.
Does ruminal metabolism of mimosine explain the absence of Leucaena toxicity in Hawaii?Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL3M3gslSgtw%3D%3D&md5=9cf51767eae9e6dd88327e78a799ba02CAS |

Jones RJ (1994) Management of anti-nutritive factors – with special reference to leucaena. In ‘Forage tree legumes in tropical agriculture’. (Eds RC Gutteridge, HM Shelton) pp. 216–231. (CAB International: Wallingford, UK)

Jones RJ, Hegarty MP (1984) The effect of different proportions of Leucaena leucocephala in the diet of cattle on growth, feed intake, thyroid function and urinary excretion of 3-hydroxy-4(1H)-pyridone. Australian Journal of Agricultural Research 35, 317–325.
The effect of different proportions of Leucaena leucocephala in the diet of cattle on growth, feed intake, thyroid function and urinary excretion of 3-hydroxy-4(1H)-pyridone.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXkvVGgsrk%3D&md5=f59ba5b0e90efa1fa8bf7aafdf68dad4CAS |

Jones RM, Jones RJ (1984) The effect of Leucaena leucocephala on liveweight gain, thyroid size and thyroxine levels of steers in south-eastern Queensland. Australian Journal of Experimental Agriculture and Animal Husbandry 24, 4–9.
The effect of Leucaena leucocephala on liveweight gain, thyroid size and thyroxine levels of steers in south-eastern Queensland.Crossref | GoogleScholarGoogle Scholar |

Jones RJ, Lowry JB (1984) Australian goats detoxify the goitrogen 3-hydroxy-4(1H) pyridone (DHP) after rumen infusion from an Indonesian goat. Experientia 40, 1435–1436.
Australian goats detoxify the goitrogen 3-hydroxy-4(1H) pyridone (DHP) after rumen infusion from an Indonesian goat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2MXlslektw%3D%3D&md5=ac4e8eee426364863c7737ece1905f06CAS |

Jones RJ, Megarrity RG (1983) Comparative toxicity responses of goats fed on Leucaena leucocephala in Australia and Hawaii. Australian Journal of Agricultural Research 34, 781–790.
Comparative toxicity responses of goats fed on Leucaena leucocephala in Australia and Hawaii.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXitVyisbs%3D&md5=f14d42a41fca215e9bcc659b08102195CAS |

Jones RJ, Megarrity RG (1986) Successful transfer of DHP-degrading bacteria from Hawaiian goats to Australian ruminants to overcome the toxicity of Leucaena. Australian Veterinary Journal 63, 259–262.
Successful transfer of DHP-degrading bacteria from Hawaiian goats to Australian ruminants to overcome the toxicity of Leucaena.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL2s%2FnsFCqsQ%3D%3D&md5=a340e7fd7190d2257b2299f99d0b31c3CAS |

Jones RJ, Blunt CG, Holmes JHG (1976) Enlarged thyroid glands in cattle grazing leucaena pastures. Tropical Grasslands 10, 113–116.

Jones RJ, Blunt CG, Nurnberg BI (1978) Toxicity of Leucaena leucocephala – effect of iodine and mineral supplements on penned steers fed a sole diet of leucaena. Australian Veterinary Journal 54, 387–392.
Toxicity of Leucaena leucocephala – effect of iodine and mineral supplements on penned steers fed a sole diet of leucaena.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaE1M%2FmtFOjtQ%3D%3D&md5=a915e406fec137f072d9443e6227ad7fCAS |

Jones RJ, Lowry JB, Megarrity RG (1985) Transfer of DHP-degrading bacteria from adapted to unadapted ruminants. Leucaena Research Reports 6, 5–6.

Jones RJ, Coates DB, Palmer B (2009) Survival of the rumen bacterium Synergistes jonesii in a herd of Droughtmaster cattle in north Queensland. Animal Production Science 49, 643–645.
Survival of the rumen bacterium Synergistes jonesii in a herd of Droughtmaster cattle in north Queensland.Crossref | GoogleScholarGoogle Scholar |

Klieve AV, Ouwerkerk D, Turner A, Roberton R (2002) The production and storage of a fermentor-grown bacterial culture containing Synergistes jonesii, for protecting cattle against mimosine and 3-hydroxy-4(1H)-pyridone toxicity from feeding on Leucaena leucocephala. Australian Journal of Agricultural Research 53, 1–5.
The production and storage of a fermentor-grown bacterial culture containing Synergistes jonesii, for protecting cattle against mimosine and 3-hydroxy-4(1H)-pyridone toxicity from feeding on Leucaena leucocephala.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhsVGrtL4%3D&md5=d7a50d6da127e6ebb3d25eccadbbac9fCAS |

Lee CP, Christie GS, Hegarty MP (1980) Anti-thyroid and anti-perioxidase activity of some isomeric dihydroxy pyrodines. Thyroid Research III. In ’Proceedings of the 8th international thyroid congress’. p. 137.

Lindsay RH, Aboul-Enein HY, Morel D, Bowen S (1974) Synthesis and antiperoxidase activity of propylthiouracil derivatives and metabolites. Journal of Pharmaceutical Sciences 63, 1383–1386.

Lowry JB, Tangendjaja B, Cook NW (1985) Measurement of mimosine and its metabolites in biological materials. Journal of the Science of Food and Agriculture 36, 799–807.
Measurement of mimosine and its metabolites in biological materials.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL28XjtlWnsA%3D%3D&md5=612c108c7393470a0dc28a36e2205894CAS |

Ludwig W, Strunk O, Westram R, Richter L, Meier H, Yadhukumar , Buchner A, Lai T, Steppi S, Jobb G, Förster W, Brettske I, Gerber S, Ginhart AW, Gross O, Grumann S, Hermann S, Jost R, König A, Liss T, Lüßmann R, May M, Nonhoff B, Reichel B, Strehlow R, Stamatakis A, Stuckmann N, Vilbig A, Lenke M, Ludwig T, Bode A, Schleifer K-H (2004) ARB: a software environment for sequence data. Nucleic Acids Research 32, 1363–1371.

Mastika IM (2003) Feeding strategies to improve the production performance and meat quality of Bali cattle (Bos sondaicus). In ‘Strategies to improve Bali cattle in Eastern Indonesia. ACIAR Proceedings No. 110’. (Eds K Entwistle, DR Lindsay) pp. 10–13. (Australian Centre for International Agricultural Research: Canberra)

Matejovic I (1995) Total nitrogen in plant material determinated by means of dry combustion: A possible alternative to determination by Kjeldahl digestion. Communications in Soil Science and Plant Analysis 26, 2217–2229.
Total nitrogen in plant material determinated by means of dry combustion: A possible alternative to determination by Kjeldahl digestion.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXntVCntrw%3D&md5=04bdd82a874fc208fcf43d6291ede781CAS |

McSweeney CS, Shelton HM (2009) Final report: effectiveness of S. jonesii inoculum for cattle grazing leucaena. Meat & Livestock Australia Limited, North Sydney, NSW.

McSweeney CS, Bamualim A, Murray RM (1984) Ruminal motility in sheep intoxicated with 2,3-dihydroxypyridine. Australian Veterinary Journal 61, 271–272.
Ruminal motility in sheep intoxicated with 2,3-dihydroxypyridine.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaL2M%2FntF2gsA%3D%3D&md5=a07fb3ac6f5593f50d130ab912742baeCAS |

McSweeney CS, Odenyo A, Krause DO (2002) Rumen microbial responses to antinutritive factors in fodder trees and shrub legumes. Journal of Applied Animal Research 21, 181–205.
Rumen microbial responses to antinutritive factors in fodder trees and shrub legumes.Crossref | GoogleScholarGoogle Scholar |

Megarrity RG, Jones RJ (1983) Toxicity of Leucaena leucocephala in ruminants – the effect of supplemental thyroxine on goats fed on a sole diet of Leucaena. Australian Journal of Agricultural Research 34, 791–798.
Toxicity of Leucaena leucocephala in ruminants – the effect of supplemental thyroxine on goats fed on a sole diet of Leucaena.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXitVyisbg%3D&md5=98769081c4629cd9f9efe7215c3c8eefCAS |

Padmanabha J, Halliday MJ, Denman SE, Davis CK, Shelton HM, McSweeney CS (2014) Is there genetic diversity in the ‘leucaena bug’ Synergistes jonesii which may reflect ability to degrade leucaena toxins? Tropical Grasslands-Forrajes Tropicales 2, 113–115.
Is there genetic diversity in the ‘leucaena bug’ Synergistes jonesii which may reflect ability to degrade leucaena toxins?Crossref | GoogleScholarGoogle Scholar |

Palmer B, Jones RJ, Poathong S, Chobtang J (2010) Within-country variation in the ability of ruminants to degrade DHP following the ingestion of Leucaena leucocephala – a Thailand experience. Tropical Animal Health and Production 42, 161–164.
Within-country variation in the ability of ruminants to degrade DHP following the ingestion of Leucaena leucocephala – a Thailand experience.Crossref | GoogleScholarGoogle Scholar |

Panjaitan T, Fauzan M, Dahlanuddin , Halliday MJ, Shelton HM (2014) Growth of Bali bulls fattened with Leucaena leucocephala in Sumbawa, Eastern Indonesia. Tropical Grasslands-Forrajes Tropicales 2, 116–118.
Growth of Bali bulls fattened with Leucaena leucocephala in Sumbawa, Eastern Indonesia.Crossref | GoogleScholarGoogle Scholar |

Pattanaik AK, Khan SA, Goswami TK (2007) Influence of iodine on nutritional, metabolic and immunological response of goats fed Leucaena leucocephala leaf meal diet. The Journal of Agricultural Science 145, 395–405.
Influence of iodine on nutritional, metabolic and immunological response of goats fed Leucaena leucocephala leaf meal diet.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXptVSgu78%3D&md5=5bdf648a82e39ce9c44dabddab9fc26dCAS |

Paul SS (2000) Detoxification of mimosine and dihydroxypyridone: a review. Agricultural Reviews (Karnal) 21, 104–109.

Phaikaew C, Suksaran W, Ted-Arsen J, Nakamanee G, Saichuer A, Seejundee S, Kotprom N, Shelton HM (2012) Incidence of subclinical toxicity in goats and dairy cows consuming leucaena (Leucaena leucocephala) in Thailand. Animal Production Science 52, 283–286.
Incidence of subclinical toxicity in goats and dairy cows consuming leucaena (Leucaena leucocephala) in Thailand.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XktFWgs7c%3D&md5=b2797a90f9871ab782ea45b7ddab81d9CAS |

Pratchett D, Jones RJ, Syrch FX (1991) Use of DHP-degrading rumen bacteria to overcome toxicity in cattle grazing irrigated leucaena pasture. Tropical Grasslands 25, 268–274.

Puchała R, Sahlu T, Davis JJ, Hart SP (1995) Influence of mineral supplementation on 2,3-dihydroxypyridine toxicity in Angora goats. Animal Feed Science and Technology 55, 253–262.
Influence of mineral supplementation on 2,3-dihydroxypyridine toxicity in Angora goats.Crossref | GoogleScholarGoogle Scholar |

Quirk MF, Bushell JJ, Jones RJ, Megarrity RG, Butler KL (1988) Live-weight gains on leucaena and native grass pastures after dosing cattle with rumen bacteria capable of degrading DHP, a ruminal metabolite from leucaena. Journal of Agricultural Science, Cambridge 111, 165–170.
Live-weight gains on leucaena and native grass pastures after dosing cattle with rumen bacteria capable of degrading DHP, a ruminal metabolite from leucaena.Crossref | GoogleScholarGoogle Scholar |

Rincón MT, Domínguez-Bello MG, Lovera M, Romero R (2000) Degradation of toxic piridinediols derived from mimosine by rumen bacteria: I. Microbiological aspects. Revista Cientifica — Facultad De Ciencias Veterinarias 10, 222–232.

Shelton HM, Brewbaker JL (1994) Leucaena leucocephala – the most widely used forage tree legume. In ‘Forage tree legumes in tropical agriculture’. (Eds RC Gutteridge, HM Shelton) pp. 15–29. (CAB International: Wallingford, UK)

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28, 2731–2739.
MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXht1eiu73K&md5=5044b1de86e06bbb6f3d1a6dc14b38afCAS |

Tan P, Wang X, Wang J (1994) Rumen bacteria degrading toxic mimosine and dihydroxypyridine compounds in China. Wei Sheng Wu Xue Bao 34, 379–384.

Tangendjaja B, Wills RBH (1980) Analysis of mimosine and 3-hydroxy-4(1H)-pyridone by high-performance liquid chromatography. Journal of Chromatography. A 202, 317–318.
Analysis of mimosine and 3-hydroxy-4(1H)-pyridone by high-performance liquid chromatography.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL3MXhsFaktb8%3D&md5=b9116a1fd06ba3e615bda7978eaa1bbcCAS |

Van Soest PJ, Robertson JB, Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74, 3583–3597.
Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK38%2FnvVCltA%3D%3D&md5=1bbe217b400e9f7fff04f7f3e179f03aCAS |

Wildin JH (1994) Beef production from broadacre leucaena in central Queensland. In ‘Forage tree legumes in tropical agriculture’. (Eds RC Gutteridge, HM Shelton) pp. 352–356. (CAB International: Wallingford, UK)