Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation and characterization of a novel gossypol-degrading bacteria Bacillus subtilis strain Rumen Bacillus Subtilis

  • Zhang, Yunhua (Anhui Province Key Laboratory of Farmland Ecological Conservation and Pollution Prevention, School of Resources and Environment, Anhui Agricultural University) ;
  • Zhang, Zhengyou (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Dai, Li (Anhui Province Key Laboratory of Farmland Ecological Conservation and Pollution Prevention, School of Resources and Environment, Anhui Agricultural University) ;
  • Liu, Ying (Anhui Province Key Laboratory of Farmland Ecological Conservation and Pollution Prevention, School of Resources and Environment, Anhui Agricultural University) ;
  • Cheng, Maoji (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Chen, Lijuan (College of Animal Science and Technology, Anhui Agricultural University)
  • Received : 2017.01.10
  • Accepted : 2017.06.19
  • Published : 2018.01.01
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Keywords

References

  1. Zhu DW, Liu ZP, Cai GL, et al. Screening of gossypol--degrading strain and its application in fermentation of cottonseed meal. China Oils Fats 2010;35:24-8.
  2. Nagalakshmi D, Rao SVR, Panda AK, Sastry VRB. Cottonseed meal in poultry diets. A review. J Poult Sci 2007;44:119-34. https://doi.org/10.2141/jpsa.44.119
  3. Santos JEP, Villasenor M, Robinson PH, Depeters EJ, Holmberg CA. Type of cottonseed and level of gossypol in diets of lactating dairy cows: plasma gossypol, health and reproductive performance. J Dairy Sci 2003;86:892-905. https://doi.org/10.3168/jds.S0022-0302(03)73672-8
  4. Carruthers NJ, Dowd MK, Stemmer PM. Gossypol inhibits calcineurin phosphatase activity at multiple sites. Eur J Pharm Sci 2007;555:106-14.
  5. Nguyen TN, Davis DA, Saoud IP. Evaluation of alternative protein sources to replace fish meal in practical diets for juvenile tilapia, Oreochromis spp. J World Aquacult Soc 2009;40:113-21. https://doi.org/10.1111/j.1749-7345.2008.00230.x
  6. El-Sharaky AS, Newairy AA, Elguindy NM, Elwafa AA. Spermatotoxicity, biochemical changes and histological alteration induced by gossypol in testicular and hepatic tissues of male rats. Food Chem Toxicol 2010;48:3354-61. https://doi.org/10.1016/j.fct.2010.09.004
  7. Tang JW, Sun H, Yao XH, et al. Effects of replacement of soybean meal by fermented cottonseed meal on growth performance, serum biochemical parameters and immune function of yellow-feathered broilers. Asian-Australas J Anim Sci 2012;25:393-400. https://doi.org/10.5713/ajas.2011.11381
  8. Pons WA, Eaves PH. Aqueous acetone extraction of cottonseed. J Am Oil Chem Soc 1967;44:460-4. https://doi.org/10.1007/BF02666795
  9. Gardner HK, Hron RJ, Vix HLE. Removal of pigment glands (gossypol) from cottonseed. Cereal Chem 1976;53:549-60.
  10. Barraza ML, Coppock CE, Brooks KN, et al. Iron sulfate and feed pelleting to detoxify FG in cottonseed diets for dairy cattle. J Dairy Sci 1991;74:3457-67. https://doi.org/10.3168/jds.S0022-0302(91)78536-6
  11. Tabatabai F, Golian A, Salarmoeini M. Determination and detoxification methods of cottonseed meal gossypol for broiler chicken rations. J Agric Sci Technol Iran 2002;16:3-15.
  12. Nagalakshmi D, Sastry VRB, Pawde A. Rumen fermentation patterns and nutrient digestion in lambs fed cottonseed meal supplemental diets. Anim Feed Sci Technol 2003;103:1-14. https://doi.org/10.1016/S0377-8401(02)00140-2
  13. Weng XY, Sun JY. Biodegradation of free gossypol by a new strain of Candida tropicalis under SSF: effects of fermentation parameters. Process Biochem 2006;41:1663-8. https://doi.org/10.1016/j.procbio.2006.03.015
  14. Zhang WJ, Xu ZR, Sun JY, Yang X. Effect of selected fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal. J Zhejiang Univ-Sci B 2006;7:690-5. https://doi.org/10.1631/jzus.2006.B0690
  15. Zhang WJ, Xu ZR, Zhao SH, Sun JY, Yang X. Development of a microbial fermentation process for detoxification of gossypol in cottonseed meal. Anim Feed Sci Technol 2007;135:176-86. https://doi.org/10.1016/j.anifeedsci.2006.06.003
  16. Khalaf MA, Meleigy SA. Reduction of free gossypol levels in cottonseed meal by microbial treatment. Int J Agric Biol 2008;10:185-90.
  17. Yang X, Sun JY, Guo JL, Weng XY. Identification and proteomic analysis of a novel gossypol-degrading fungal strain. J Sci Food Agric 2012;92:943-51. https://doi.org/10.1002/jsfa.4675
  18. Willard ST, Neuendorff DA, Lewis AW, Randel RD. Effect of free gossypol in the diet of pregnant and postpartum Brahman cows on calf development and cow performance. Anim Sci 1995;73:496-507. https://doi.org/10.2527/1995.732496x
  19. Xie JY, Dong GJ, Liu ZY. Method of preparation of microbiological specimen for scanning electron microsope. J Chinese Electron Microsc Soc 2005;24:440.
  20. Berger SL, Kimmel AR. Guide to molecular cloning techniques. Methods Enzymol 1987;152: 33-41.
  21. Zhang Y, Luan H, Wei Z, et al. Exploiting of honey-associated Bacillus strains as plant-growth promoting bacteria for enhancing barley growth in rare earth tailings. Ann Microbiol 2016;66:559-68.
  22. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406-25.
  23. Tamura K, Dudley J, Nei M, Kumar S. Mega4: molecular evolutionary gnentics analysis (MEGA) software version 4.0. Mol Biol Evol 2007;24:1596-9. https://doi.org/10.1093/molbev/msm092
  24. AOAC. Official methods of analysis. 16th ed. Washington, DC, USA:AOAC International; 1999.
  25. Deruiter J, Jacyno JM, Davis RA, Cutler HG. Studies on aldose reductase inhibitors from fungi. I Citrinin and related benzopyran derivatives. J Enzyme Inhib 1992;6:201-10. https://doi.org/10.3109/14756369209020170
  26. Mazza P. The use of Bacillus tequliensis as an antidiarrhoeal microorganism. Boll Chim Farm 1994;133:3-18.
  27. Romero-Garcia S, Hernandez-Bustos C, Merino E, Gosset G, Martinez A. Homolactic fermentation from glucose and cellobiose using Bacillus tequliensis. Microb Cell Fact 2009;8:23.
  28. Rajesh K, Subhas CM, Kurcheti PP, Asim KP. Evaluation of Bacillus tequliensis as a probiotic to Indian major carp Labeo rohita (Ham). Aquac Res 2006;37:1215-21.
  29. Botsoglou NA. Determination of "free" gossypol in cottonseed and cottonseed meals by second-derivative ultraviolet spectrophotometry. J Agric Food Chem 1991;39:478-82. https://doi.org/10.1021/jf00003a010
  30. Smith FH. Determination of gossypol in leaves and flower buds of Gossypium. J Am Oil Chem Soc 1971;44:267-9.
  31. Lee KJ, Konrad D. High-performance liquid chromatographic determination of gossypol and gossypolone enantiomers in fish tissues using simultaneous electrochemical and ultraviolet detectors. J Chromatogr B 2002;779:313-9. https://doi.org/10.1016/S1570-0232(02)00402-6
  32. Dong SL, Hong JM. Determination of free gossypol by AAS. J Chinese Public Health 1993;9:366-7.
  33. Benbouza H, Lognay G, Palm R, Baudoin JP, Mergeai G. Development of a visual method to quantify the gossypol content in cottonseeds. Crop Sci 2002;42:1937-42. https://doi.org/10.2135/cropsci2002.1937
  34. Cai YF, Zhang H, Zeng Y, et al. An optimized gossypol high performance liquid chromatography assay and its application in evaluation of different gland genotypes of cotton. J Biosci 2004;29:67-71. https://doi.org/10.1007/BF02702563
  35. Wu WW, Xu GR. Determination of free gossypol in cottonseed cake feed by HPLC. J Agric Univ Hebei 2009;32:105-8.
  36. Kamga R, Kayem GJ, Rouxhe PG. Adsorption of gossypol from cottonseed oil on oxides. J Colloid Interface Sci 2000;232:198-206. https://doi.org/10.1006/jcis.2000.7191
  37. Rhee KS, Ziprin YA, Calhoun MC. Antioxidative effects of cottonseed meals as evaluated in cooked meat. Meat Sci 2001;58:117-23. https://doi.org/10.1016/S0309-1740(00)00137-6
  38. Abou-Donia MB, Dieckert JW. Metabolic fate of gossypol: The metabolism of [14C] gossypol in swine. Toxico Appl Pharm 1975;31:32-46. https://doi.org/10.1016/0041-008X(75)90049-6
  39. Schneider IC, Ames ML, Rasmussen MA. Fermentation of cottonseed and other feedstuffs in cattle rumen fluid. J Agric Food Chem 2002; 50:2267-73.

Cited by

  1. Secretome Analysis and Bioprospecting of Lignocellulolytic Fungal Consortium for Valorization of Waste Cottonseed Cake by Hydrolase Production and Simultaneous Gossypol Degradation pp.1877-265X, 2019, https://doi.org/10.1007/s12649-019-00620-1
  2. Impacts of Dietary Protein from Fermented Cottonseed Meal on Lipid Metabolism and Metabolomic Profiling in the Serum of Broilers vol.21, pp.8, 2020, https://doi.org/10.2174/1389203721666200203152643
  3. Isolation and Identification of a Rumen Lactobacillus Bacteria and Its Degradation Potential of Gossypol in Cottonseed Meal during Solid-State Fermentation vol.9, pp.11, 2018, https://doi.org/10.3390/microorganisms9112200
  4. Gossypol detoxification in the rumen and Helicoverpa armigera larvae: A review vol.7, pp.4, 2021, https://doi.org/10.1016/j.aninu.2021.02.007
  5. Enhancing the Quality of Total Mixed Ration Containing Cottonseed or Rapeseed Meal by Optimization of Fermentation Conditions vol.7, pp.4, 2018, https://doi.org/10.3390/fermentation7040234