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Journal of Integrative Agriculture  2020, Vol. 19 Issue (6): 1644-1655    DOI: 10.1016/S2095-3119(20)63174-4
Special Issue: 动物营养合辑Animal Nutrition
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Effects of different molecular weights of chitosan on methane production and bacterial community structure in vitro
TONG Jin-jin1, 2*, ZHANG Hua1*, WANG Jia1, LIU Yun3, MAO Sheng-yong4, XIONG Ben-hai5, JIANG Lin-shu1 
1 Beijing Key Laboratory for Dairy Cow Nutrition, Beijing University of Agriculture, Beijing 102206, P.R.China
2 Beijing Bei Nong Enterprise Management Co., Ltd., Beijing 102206, P.R.China
3 College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P.R.China
4 College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China
5 State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
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Abstract  
As a new feed additive, chitosan has been shown in recent years to have a certain role in reducing methane emissions from the gastrointestinal tracts of ruminants.  However, the effects of chitosan with different molecular weights on rumen fermentation, methane production and bacterial community structure are not yet clear.  A basal diet without chitosan served as the control (CTL), and the treatment diets were supplemented with chitosan with different molecular weights: 1 000 (1K), 3 000 (3K), 5 000 (5K), 50 000 (5W) and 200 000 (20W) dry matter (DM).  Six fermentation units per treatment were established.  Gas chromatography was used to measure the concentrations of methane, H2 and volatile fatty acids (VFAs).  The bacterial 16S rRNA genes were sequenced with an Illumina MiSeq platform and analysed to reveal the relative abundances of bacterial community taxa.  The results showed that the propionate proportion was significantly increased by the addition of chitosan with different molecular weights (P<0.05), while methane production and the acetate proportion were significantly decreased (P<0.05).  The relative abundances of Rikenellaceae_RC9_gut_group and Prevotellaceae_UCG_003 were significantly increased in the 3K chitosan group compared with the CTL group, whereas the relative abundance of Ruminococcaceae_NK4A214_group was significantly decreased (P<0.05).  Correlation analyses of the relative abundances of the bacterial genera showed that Prevotella was positively related to propionate production (P<0.05).  In conclusion, 3K chitosan could reduce methane production by replacing fibrolytic bacteria (Firmicutes and Fibrobacteres) with amylolytic bacteria (Bacteroidetes and Proteobacteria) in the bacterial community structure.
 
Keywords:  chitosan        high-throughput DNA sequencing        methane  
Received: 27 February 2019   Accepted:
Fund: This study was financially supported by the projects of the National Natural Science Foundation of China (31802091, 31702302 and 31772629) and the National Key Research and Development Plan of China (2016YFD0700205, 2016YFD0700201 and 2017YFD0701604). 
Corresponding Authors:  Correspondence JIANG Lin-shu, Mobile: +86-13801059171, Fax: +86-10-80796368, E-mail: jls@bac.edu.cn; XIONG Ben-hai, Mobile: +86-13801090939, Fax: +86-10-62815988, E-mail: xiongbenhai@caas.cn   
About author:  * These authors contributed equally to this study.

Cite this article: 

TONG Jin-jin, ZHANG Hua, WANG Jia, LIU Yun, MAO Sheng-yong, XIONG Ben-hai, JIANG Lin-shu. 2020. Effects of different molecular weights of chitosan on methane production and bacterial community structure in vitro. Journal of Integrative Agriculture, 19(6): 1644-1655.

Araújo A P C, Venturelli B C, Santos M C B, Gardinal R, Cônsolo N R B, Calomeni G D, Freitas J E, Barletta R V, Gandra J R, Paiva P G. 2015. Chitosan affects total nutrient digestion and ruminal fermentation in Nellore steers. Journal of Animal Feed Science and Technology, 206, 114–118.
Asli A, Brouillette E, Ster C, Ghinet M G, Brzezinski R, Lacasse P, Jacques M, Malouin F. 2017. Antibiofilm and antibacterial effects of specific chitosan molecules on Staphylococcus aureus isolates associated with bovine mastitis. PLoS ONE, 12, e0176988.
Avgustin G, Wallace R J, Flint H J. 1997. Phenotypic diversity among ruminal isolates of Prevotella ruminicola: Proposal of Prevotella brevis sp. nov., Prevotella bryantii sp. nov., and Prevotella albensis sp. nov. and redefinition of Prevotella ruminicola. International Journal of Systematic Bacteriology, 47, 284–288.
Belanche A, Pinloche E, Preskett D, Newbold C J. 2016. Effects and mode of action of chitosan and ivy fruit saponins on the microbiome, fermentation and methanogenesis in the rumen simulation technique. FEMS Microbiology Ecology, 92, 1–12.
Belanche A, Ramos-Morales E, Newbold C J. 2015. In vitro screening of natural feed additives from crustaceans, diatoms, seaweeds and plant extracts to manipulate rumen fermentation. Journal of the Science of Food & Agriculture, 96, 3069–3078.
Broucek J, Uhrincat M, Soch M, Lendelova J, Hanus A, Stastna J, Botto L. 2013. Effect of management change on selected welfare parameters of cows. Animal Science Papers and Reports, 31, 195–203.
Chaney A L, Marbach E P. 1962. Modified reagents for determination of urea and ammonia. Clinical Chemistry, 8, 130–132.
Cieslak A, Szumacher-Strabel M, Stochmal A, Oleszek W. 2013. Plant components with specific activities against rumen methanogens. Animal an International Journal of Animal Bioscience, 7(Suppl. 2), 253.
Danielsson R, Dicksved J, Li S, Gonda H, Müller B, Schnürer A, Bertilsson J. 2017. Methane production in dairy cows correlates with rumen methanogenic and bacterial community structure. Frontiers in Microbiology, 8, 226–238.
Firkins J L, Yu Z, Morrison M. 2007. Ruminal nitrogen metabolism: Perspectives for integration of microbiology and nutrition for dairy. Journal of Dairy Science, 90, E1.
Gill M, Smith P, Wilkinson J M, Simm G. 2010. Mitigating climate change: The role of domestic livestock. Animal, 4, 323–333.
Goiri I, Garciarodriguez A, Oregui L M. 2009a. Effect of chitosan on mixed ruminal microorganism fermentation using the rumen simulation technique (Rusitec). Animal Feed Science & Technology, 152, 92–102.
Goiri I, Garciarodriguez A, Oregui L M. 2009b. Effect of chitosans on in vitro rumen digestion and fermentation of maize silage. Animal Feed Science & Technology, 148, 276–287.
Goiri I, Oregui L M, Garcia-Rodriguez A. 2009c. Dose-response effects of chitosans on in vitro rumen digestion and fermentation of mixtures differing in forage-to-concentrate ratios. Animal Feed Science & Technology, 151, 215–227.
Goiri I, Oregui L M, Garcia-Rodriguez A. 2010. Use of chitosans to modulate ruminal fermentation of a 50:50 forage-to-concentrate diet in sheep. Journal of Animal Science, 88, 749–755.
Janssen P H. 2010. Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics. Animal Feed Science & Technology, 160, 1–22.
Joblin K N. 1999. Ruminal acetogens and their potential to lower ruminant methane emissions. Australian Journal of Agricultural Research, 50, 1307–1314.
Jones D M. 1981. Manual of methods for general bacteriology. Journal of Clinical Pathology, 34, 1069.
Kang S H, Evans P, Morrison M, Mcsweeney C. 2013. Identification of metabolically active proteobacterial and archaeal communities in the rumen by DNA- and RNA-derived 16S rRNA gene. Journal of Applied Microbiology, 115, 644–653.
Knorr D. 1991. Antimicrobial effect of water-soluble chitosans with high hydrostatic pressure. Food Biotechnology, 5, 45–57.
Lanctôt S, Fustier P, Taherian A R, Bisakowski B, Zhao X, Lacasse P. 2017. Effect of intramammary infusion of chitosan hydrogels at drying-off on bovine mammary gland involution. Journal of Dairy Science, 100, 2269–2281.
Leahy S C, Kelly W J, Altermann E, Ronimus R S, Yeoman C J, Pacheco D M, Li D, Kong Z, Mctavish S, Sang C. 2010. The genome sequence of the rumen methanogen Methanobrevibacter ruminantium reveals new possibilities for controlling ruminant methane emissions. PLoS ONE, 5, e8926.
Leahy S C, Kelly W J, Ronimus R S, Wedlock N, Altermann E, Attwood G T. 2013. Genome sequencing of rumen bacteria and archaea and its application to methane mitigation strategies. Animal, 7, 235–243.
Mao S Y, Zhang G, Zhu W Y. 2008. Effect of disodium fumarate on ruminal metabolism and rumen bacterial communities as revealed by denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA. Animal Feed Science & Technology, 140, 293–306.
Martin C, Rouel J, Jouany J P, Doreau M, Chilliard Y. 2008. Methane output and diet digestibility in response to feeding dairy cows crude linseed, extruded linseed, or linseed oil. Journal of Animal Science, 86, 2642–2650.
Martinez-Fernandez G, Denman S E, Yang C, Cheung J, Mitsumori M, Mcsweeney C S. 2016. Methane inhibition alters the microbial community, hydrogen flow, and fermentation response in the rumen of cattle. Frontiers in Microbiology, 7, 1087–1099.
May T, Rychlik J L. 2000. The effect of a methanogen, methanobrevibacter smithii, on the growth rate, organic acid production, and specific ATP activity of three predominant ruminal cellulolytic bacteria. Current Microbiology, 40, 176–180.
Mima S, Miya M, Iwamoto R, Yoshikawa S. 1983. Highly deacetylated chitosan and its properties. Journal of Applied Polymer Science, 28, 1909–1917.
Ming K, Chen X G, Ke X, Hyunjin P. 2010. Antimicrobial properties of chitosan and mode of action: A state of the art review. International Journal of Food Microbiology, 144, 51–63.
Mitsumori M, Shinkai T, Takenaka A, Enishi O, Higuchi K, Kobayashi K, Nonaka I, Asanuma N, Denman S E, Mcsweeney C S. 2012. Responses in digestion, rumen fermentation and microbial populations to inhibition of methane formation by a halogenated methane analogue. British Journal of Nutrition, 108, 482–491.
Morgavi D P, Forano E, Martin C, Newbold C J. 2010. Microbial ecosystem and methanogenesis in ruminants. Animal an International Journal of Animal Bioscience, 4, 1024–1036.
Negussie E, De H Y, Dehareng F, Dewhurst R J, Dijkstra J, Gengler N, Morgavi D P, Soyeurt H, Van G S, Yan T. 2017. Invited review: Large-scale indirect measurements for enteric methane emissions in dairy cattle: A review of proxies and their potential for use in management and breeding decisions. Journal of Dairy Science, 100, 2433–2453.
NRC (National Research Council). 2001. Nutrient Requirements of Dairy Cattle. 7th ed. National Academies Press, Washington, D.C.
Paiva P G D, Jesus E F D, Valle T A D, Almeida G F D, Consentini C E C, Zanferari F, Takiya C F, Bueno I C, Renn F P. 2016. Effects of chitosan on ruminal fermentation, nutrient digestibility, and milk yield and composition of dairy cows. Animal Production Science, 57, 136–142.
Patra T, Pal A, Dey J. 2010. A smart supramolecular hydrogel of Nα-(4-n-Alkyloxybenzoyl)-l-histidine exhibiting pH-modulated properties. Langmuir, 26, 7761–7767.
Rhoades J, Roller S. 2000. Antimicrobial actions of degraded and native chitosan against spoilage organisms in laboratory media and foods. Applied & Environmental Microbiology, 66, 80–92.
Russel J B. 1998. The importance of pH in the regulation of ruminal acetate to propionate ratio and methane production in vitro. Journal of dairy science, 81, 3222–3230.
SSTCC (The State Science and Technology Commission of the P.R.China). 2017. Regulation on the Administration of Laboratory Animals (2017 revision). National Science and Technology Council of the People’s Republic of China. (in Chinese)
Stewart C, Flint H, Bryant M. 1997. The rumen bacteria. In: Hobson P G, Stewart C S, eds., The Rumen Microbial Ecosystem. Chapman and Hall, London, UK. pp. 10–72.
Sudarshan N R, Hoover D G, Knorr D. 2009. Antibacterial action of chitosan. Food Biotechnology, 6, 257–272.
Tanja M, Salzberg S L. 2011. FLASH: Fast length adjustment of short reads to improve genome assemblies. Bioinformatics, 27, 2957–2963.
Terbojevich M, Cosani A, Focher B, Marsano E. 1993. High-performance gel-permeation chromatography of chitosan samples. Carbohydrate Research, 250, 301–314.
Thao N T, Wanapat M, Cherdthong A, Kang S. 2014. Effects of eucalyptus crude oils supplementation on rumen fermentation, microorganism and nutrient digestibility in swamp buffaloes. Asian-Australasian Journal of Animal Sciences, 27, 46–54.
Tong J J, Zhang H, Yang D L, Zhang Y H, Xiong B H, Jiang L S. 2018. Illumina sequencing analysis of the ruminal microbiota in high-yield and low-yield lactating dairy cows. PLoS ONE, 13, e0198225.
Ungerfeld E M. 2015. Shifts in metabolic hydrogen sinks in the methanogenesis-inhibited ruminal fermentation: A meta-analysis. Frontiers in Microbiology, 6, 37–49.
Vargas J E, Andrés S, Snelling T J. 2017. Effect of sunflower and marine oils on ruminal microbiota, in vitro fermentation and digesta fatty acid profile. Frontiers in Microbiology, 8, 1124–1136.
Wang K, Nan X M, Chu K K, Tong J J, Yang L, Zheng S S, Zhao G, Jiang L S, Xiong B H. 2018. Shifts of hydrogen metabolism from methanogenesis to propionate production in response to replacement of forage fiber with non-forage fiber sources in diets in vitro. Frontiers in Microbiology, 9, 2764–2776.
Weller R A, Pilgrim A F. 1974. Passage of protozoa and volatile fatty acids from the rumen of the sheep and from a continuous in vitro fermentation system. British Journal of Nutrition, 32, 341–351.
Whitelaw F G, Eadie J M, Bruce L A, Shand W J. 1984. Methane formation in faunated and ciliate-free cattle and its relationship with rumen volatile fatty acid proportions. British Journal of Nutrition, 52, 261.
Zened A, Combes S, Cauquil L, Mariette J, Klopp C, Bouchez O, Troegeler-Meynadier A, Enjalbert F. 2013. Microbial ecology of the rumen evaluated by 454 GS FLX pyrosequencing is affected by starch and oil supplementation of diets. FEMS Microbiology Ecology, 83, 504–514.
Zhang J, Tong J J, Jiang L S. 2018. Effects of different molecular weights and different concentrations of chitosan on rumen in vitro fermentation parameters and methane emission in dairy cows. Animal Nutrition, 30, 458–466. (in Chinese)
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