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Recent advances in rumen manipulation
techniques with particular reference to
         ruminant production




          By : Tetemke Kidane
INTRODUCTION
• Why do we need to manipulate the rumen?
• 15 to 20% of GEI is lost in the form of methane
  and heat due to rumen fermentation
• 8 -10 % of dietary ME lost as methane -
  methanogenic bacteria
• Amino acids are hydrolyzed and their constituent
  amino acids deaminated by microbes
• manipulate the microbial population to minimize
  the degradation of feed protein
Cont’d…

• use of chemicals inhibit the activity of
microbial proteases or deaminases
            treatment of feedstuffs
                   inhibit ruminal proteolysis
              various drying procedures, heat
 Formaldehyde-treated feeds - increases
efficiency of growth
 use of some proteins to coat and protect fat
       • To enhance yields of milk and to increase amounts
         of USFA in milk or animal fat.
The digestive tract of a health adult ruminant




(Source: Russell, 2000)
The rumen microbes
• Bacteria- gram-positive and gram-negative
     • most numerous group of microbes in the rumen
     • Substrates are fermented to form volatile fatty acids
       (Acetate, Propionate, Butyrate and others) and C02
       and CH4
• Protozoa
     • Generally larger than bacteria but present in small no.
     • Fibre digestion
     • Negatively influence protein utilization – predation
       and reduce outflow of microbial protein
     • Ruminants can survive with out protozoa
• Fungi
  – recently discovered ruminal MOs
  – Degrade the lignin-containing tissues than bacteria
livestock agriculture’s role in greenhouse
                       gases
                         CH4                        % of
                         million metric      % of
   anthropogenic
Sources                  tons/yr          total     sources
Natural
 Wetland                   115            24.4         -
 Oceans                     15             3.2         -
 Termites                   20             4.2         -
 Burning                    10             2.1         -
Industrial
 Gas and oil                   50          10.6       16.1
 Coal                          40           8.4       12.8
 Charcoal                      10           2.1        3.2
 Landfills                     30           6.4        9.6
 Waste water treatment         25           5.3        8.0
Agricultural
 Rice                        60             12.8       19.3
 Livestock                   80             17.0        25.8
 Manure                     10-25            2.1       3.2-7.7
 Burning Anon, 2006
   Source:                    5               1.0        1.6
Mitigation of methane from livestock
•   Ruminal fermentation time
•   Alternate hydrogen acceptors
•   Use of feed additives
•   Improvement in production efficiency
    – Any practice that increases productivity per
      animal reduces methane emissions
• Enhancing ruminal acetogens
    – Acetogens are a group of rumen microbes
      that produce acetic acid from hydrogen and
      carbon dioxide rather than methane
• Modification of bacteria in the rumen
Fig. Brief outline of the pathways of carbohydrate
             fermentation in the rumen




   (Source: Preston and Leng, 1986)
Effect of improving the efficiency of rumen fermentative activity in
                                                            methane production


                            Effect of supplementation on methane production                                     Effect of bypass protein and other supplements on ruminal
                                                                                                                                    methane production
                            20             No
                                       supplemen
                                         tation                                                                 1200
                            15
% DE fermented to methane




                                                   Urea/miner                                                                                               No supplementation
                                                       al
                                                   supplemen
                                                                                                                 800
                            10
                                                       ts




                                                                              Methane production (Kg/ton Lwt.
                                                                                                                                                            Urea/mineral +



                                                                                                      gain)
                                                                                                                                                            bypass protein
                             5                                                                                   400                                        supplements


                             0
                                                                                                                    0
                                                  1
                                                                                                                                      1
                                    ( Source: Leng, 1991)
Effect of improving rumen fermentative activity in methane production




( Source: Leng, 1991)
Manipulation of R F with Organic Acids Supplementation

• through treatment of roughage, concentrate and
  strategic supplementation with organic acids
• improve rumen efficiency by maintaining higher
  pH, optimum ammonia-nitrogen (NH3-N), thus
  CH4 and increasing microbial protein synthesis
  and essential VFAs
• Recently, there has been increased public scrutiny
  about use of antibiotic feed additives in food
  animal production, especially in Europe
• Use of organic acids, non-antibiotic feed additives
  may alleviate public skepticism
Cont’d…
• Organic acids (OAs) have effects on
  ruminal fermentation analogous to
  ionophores (CH4 , lactate, and propionate)
• However, the mode of action for the organic
  acids is different than ionophores
• Organic acids stimulate rather than inhibit
  some specific ruminal bacterial populations
• Organic acids that are currently being
  evaluated as feed additives are Malic acid,
  Fumaric acid, and Aspartic acid
Cont’d…
• Recent research showed that a combination of
  organic acids ( malate) and monensin
        – more effective at reducing lactate
          concentrations
        – and increasing pH in mixed ruminal
          microorganism fermentation than the
          addition of organic acid or monensin
          alone
in vitro studies - response to supplemental malic acid


                                    Response to supplementation
 Culture
                  Treat
   system                     PH    TVFA   Ac      Pr    Bu     Lac    CH     NH3-
                                                                            4   N

 Batch (steer)a   0,4,8,12           NE      NE     NE                 NE
                      mM

 RUSTIEC          0,5.62       NE            NE           NE            -
   (sheep)b           mmol

 Batch (sheep)c   0,4,7,10                                              -      -
                      mM

 RUSTIEC          0,6.55       NE            NE                   -    NE      -
   (sheep)d           mM

 Continuous       0,50,100     NE    NE      NE     NE    NE           NE     NE
    (dairy            g/h/d
    cows)e

 Up arrow = significant (P<0.01), NE= No effect (P<0.01), a,b,c,d &e= References
Effect of supplemental malic acid in dairy cattle (in vivo
   studies)
                               Response to supplementation
                      DMI     Milk      milk      R.         R.      R.
 Animal       Treat           yield     fat       PH         VFA     CH
                                                                          4 References

Dairy
    cattl
    e
Holstein    0,28 ,                          -       -          -      -     Stallcup, 1979
                70g
Holstein    0,100g                                  -          -      -     Stallcup, 1979


Holstein    10,20 g                                                         Khampa et
                                                                               al.,2006
Holstein    0, 4 g      NE       NE        NE      NE        A,B,P    -     Vicini et al.,
                                                                                2003
Holstein    0,84 g                         NE       -                 -     Devan & Bach,
                                                                               2004

Up arrow = significant (P<0.01), NE= No effect (P<0.01),
Effect of supplemental malic acid in beef cattle and small ruminants

                                Response to supplementation               Reference
                      DMI   Milk      milk Gai      R. R R.         R.    s
 Animal      Treat                    com n         PH . NH         CH4
                                      p                V 3-N
                                                       F
                                                       A
Crossbred 0, 40,      NE         -        -             -   -   -    -    Martin et
steers    80 g                                                            al.1999
Angle       0,60,12   NE         -        -    NE       -   -   -    -    Martin et
steers      0g                                                            al.1999
Beef        0, 100g   NE         -        -    NE   -       -   -    -    Martin et
cattle                                                                    al.1999
HF steers   0,9,18,              -        -    -            -       NE    Khampa et
            27g                                                           al., 2006
Dairy       0,        NE        NE        NE   -        -   -   -    -    Salama et
goats       0.32%                                                         al. 2002
Lambs       0, 0.2%         -         -                     -   -    -    Flores et
                                                                          al.2003

     Up arrow = significant (P<0.01), NE= No effect (P<0.01),
Manipulation of rumen with Ionophores
• Ionophores are antibiotics includes: monensin,
  lasalocid, tetronasin, salinomycin, lysocellin,
  narasin, nigericin, laidlomycin and valynomycin
• Since the mid-1970’s they have been
  extensively used to manipulate rumen
  fermentation
• ionophores are approved in several countries
  including Australia, Argentina, New Zealand, and
  South Africa and recently USA
• Monensin (trade name Rumensin) widely used
  ionophores
Cont’d…
• Effect of ionophores on rumen end products
      • decrease in hydrogen, a precursor of methane,
      • Favors propionate production
      • Ionophores inhibit gram-positive bacteria
      • major benefit of feeding ionophores to dairy cattle is the
        potential increase in production of propionate and the
        associated decrease in production of methane, which
        conserves energy
•
Safety (residual effect)
• Several researchers indicated that meat
  and milk produced from animals fed
  monensin is safe for human consumption
• Likewise, monensin is biodegradable in
  manure and soil, and is not toxic for crop
  plants
The effects of ionophores on fermentation end products and total ciliate
                 protozoal populations in Angus yearling steers
•CC= control
•CM=33 mg/kg Monensin
• CM/L= Biweekly rotation
of monensin
(33 mg/kg) and lasalocid
(36 mg/kg)




                                                     (Source: Guan et al. 2005)
Manipulation of ruminal
 fermentation by plant extracts
• Plant extracts have been used for
  centuries for various purposes (as
  traditional medicine and food
  preservatives, among others
• The antimicrobial activity of plant extracts -
  secondary plant metabolites (saponins,
  tannins, and essential oils)
• selective inhibitors of methanogens
Cont’d…
• Plants exhibiting anti-methanogenic
  activity include: Equisetum arvense, Lotus
  corniculatus, Rheum palmatum, Salvia
  officinalis, Sapindus saponaria, Uncaria
  gambir and Yucca schidigera
• Major commercial source of saponins is
  Yucca schidiger
• Defaunating agent and selective inhibitor-
  gram positive bacteria
Effect of Yucca Schidigera extract ( Saponins) in vitro ruminal
                  characteristics and gas production
                     Yucca schidigera extract (ml/l)
                                                               SEM    P value
                     0           2             4       6
pH                   6.76        6.79          6.74    6.72    0.02   NS
NH3-N (mg/l)         28.60       21.15         18.19   14.86   1.61   ***
TVFA(mM)             66.45       64.22         67.97   69.24   1.57   NS

VFA (mM)
     Acetate (A)     47.71       44.12         45.50   46.26   1.05   NS

Propionate (P)       12.37       14.20         17.07   19.02   0.49   ***
      Butyrate       5.69        5.54          4.92    3.64    0.19   ***

      Others         0.67        0.35          0.49    0.32    0.11   NS

A:P ratio            4.42        3.88          3.61    3.52    0.06   ***

Protozoa (x104/ml)   13.16       9.38          5.90    5.80    1.07   ***

CH4 (ml/min)         0.24        0.20          0.17    0.14    0.00   ***

CO2 (ml/min)         1.22        1.21          1.39    1.56    0.06   ***

  ( Source: Pen, 2007)
Conclusions


• Rumen manipulation has paramount importance in
  improving ruminal fermentation efficiency and
  productivity in dairy and fattening ruminants in the
  tropics where the major sources of the feed is high
  in fibre.

• Rumensin from ionophores and Malic acid from
  organic acid are the widely used rumen modifiers in
  both dairy and fattening ruminant animals
Cont’d…
• Several literatures reported that use of one type
  of manipulating technique for prolonged time is
  not effective because of adaptation by the
  rumen microbes, hence, alternative or rotational
  use rather than one alone may be prolonged its
  effect in improving the performance of the
  animal
• The studies conducted with plant extracts are limited to
  in vitro screening for the desired characteristics and only
  a few of them have been tested in in vivo experiments
Cont’d…

• it is desirable to standardize in vivo dose of
  these plant extracts which are effective in
  inhibiting methane emission with minimum
  adverse effects on fermentation of feed so that
  these can be practically exploited for economic
  and ‘ecologically friendly’ livestock production
Thank You for your kind attention

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Rumen manupilation

  • 1. Recent advances in rumen manipulation techniques with particular reference to ruminant production By : Tetemke Kidane
  • 2. INTRODUCTION • Why do we need to manipulate the rumen? • 15 to 20% of GEI is lost in the form of methane and heat due to rumen fermentation • 8 -10 % of dietary ME lost as methane - methanogenic bacteria • Amino acids are hydrolyzed and their constituent amino acids deaminated by microbes • manipulate the microbial population to minimize the degradation of feed protein
  • 3. Cont’d… • use of chemicals inhibit the activity of microbial proteases or deaminases treatment of feedstuffs inhibit ruminal proteolysis various drying procedures, heat  Formaldehyde-treated feeds - increases efficiency of growth  use of some proteins to coat and protect fat • To enhance yields of milk and to increase amounts of USFA in milk or animal fat.
  • 4. The digestive tract of a health adult ruminant (Source: Russell, 2000)
  • 5. The rumen microbes • Bacteria- gram-positive and gram-negative • most numerous group of microbes in the rumen • Substrates are fermented to form volatile fatty acids (Acetate, Propionate, Butyrate and others) and C02 and CH4 • Protozoa • Generally larger than bacteria but present in small no. • Fibre digestion • Negatively influence protein utilization – predation and reduce outflow of microbial protein • Ruminants can survive with out protozoa • Fungi – recently discovered ruminal MOs – Degrade the lignin-containing tissues than bacteria
  • 6. livestock agriculture’s role in greenhouse gases CH4 % of million metric % of anthropogenic Sources tons/yr total sources Natural Wetland 115 24.4 - Oceans 15 3.2 - Termites 20 4.2 - Burning 10 2.1 - Industrial Gas and oil 50 10.6 16.1 Coal 40 8.4 12.8 Charcoal 10 2.1 3.2 Landfills 30 6.4 9.6 Waste water treatment 25 5.3 8.0 Agricultural Rice 60 12.8 19.3 Livestock 80 17.0 25.8 Manure 10-25 2.1 3.2-7.7 Burning Anon, 2006 Source: 5 1.0 1.6
  • 7. Mitigation of methane from livestock • Ruminal fermentation time • Alternate hydrogen acceptors • Use of feed additives • Improvement in production efficiency – Any practice that increases productivity per animal reduces methane emissions • Enhancing ruminal acetogens – Acetogens are a group of rumen microbes that produce acetic acid from hydrogen and carbon dioxide rather than methane • Modification of bacteria in the rumen
  • 8. Fig. Brief outline of the pathways of carbohydrate fermentation in the rumen (Source: Preston and Leng, 1986)
  • 9. Effect of improving the efficiency of rumen fermentative activity in methane production Effect of supplementation on methane production Effect of bypass protein and other supplements on ruminal methane production 20 No supplemen tation 1200 15 % DE fermented to methane Urea/miner No supplementation al supplemen 800 10 ts Methane production (Kg/ton Lwt. Urea/mineral + gain) bypass protein 5 400 supplements 0 0 1 1 ( Source: Leng, 1991)
  • 10. Effect of improving rumen fermentative activity in methane production ( Source: Leng, 1991)
  • 11. Manipulation of R F with Organic Acids Supplementation • through treatment of roughage, concentrate and strategic supplementation with organic acids • improve rumen efficiency by maintaining higher pH, optimum ammonia-nitrogen (NH3-N), thus CH4 and increasing microbial protein synthesis and essential VFAs • Recently, there has been increased public scrutiny about use of antibiotic feed additives in food animal production, especially in Europe • Use of organic acids, non-antibiotic feed additives may alleviate public skepticism
  • 12. Cont’d… • Organic acids (OAs) have effects on ruminal fermentation analogous to ionophores (CH4 , lactate, and propionate) • However, the mode of action for the organic acids is different than ionophores • Organic acids stimulate rather than inhibit some specific ruminal bacterial populations • Organic acids that are currently being evaluated as feed additives are Malic acid, Fumaric acid, and Aspartic acid
  • 13. Cont’d… • Recent research showed that a combination of organic acids ( malate) and monensin – more effective at reducing lactate concentrations – and increasing pH in mixed ruminal microorganism fermentation than the addition of organic acid or monensin alone
  • 14. in vitro studies - response to supplemental malic acid Response to supplementation Culture Treat system PH TVFA Ac Pr Bu Lac CH NH3- 4 N Batch (steer)a 0,4,8,12 NE NE NE NE mM RUSTIEC 0,5.62 NE NE NE - (sheep)b mmol Batch (sheep)c 0,4,7,10 - - mM RUSTIEC 0,6.55 NE NE - NE - (sheep)d mM Continuous 0,50,100 NE NE NE NE NE NE NE (dairy g/h/d cows)e Up arrow = significant (P<0.01), NE= No effect (P<0.01), a,b,c,d &e= References
  • 15. Effect of supplemental malic acid in dairy cattle (in vivo studies) Response to supplementation DMI Milk milk R. R. R. Animal Treat yield fat PH VFA CH 4 References Dairy cattl e Holstein 0,28 , - - - - Stallcup, 1979 70g Holstein 0,100g - - - Stallcup, 1979 Holstein 10,20 g Khampa et al.,2006 Holstein 0, 4 g NE NE NE NE A,B,P - Vicini et al., 2003 Holstein 0,84 g NE - - Devan & Bach, 2004 Up arrow = significant (P<0.01), NE= No effect (P<0.01),
  • 16. Effect of supplemental malic acid in beef cattle and small ruminants Response to supplementation Reference DMI Milk milk Gai R. R R. R. s Animal Treat com n PH . NH CH4 p V 3-N F A Crossbred 0, 40, NE - - - - - - Martin et steers 80 g al.1999 Angle 0,60,12 NE - - NE - - - - Martin et steers 0g al.1999 Beef 0, 100g NE - - NE - - - - Martin et cattle al.1999 HF steers 0,9,18, - - - - NE Khampa et 27g al., 2006 Dairy 0, NE NE NE - - - - - Salama et goats 0.32% al. 2002 Lambs 0, 0.2% - - - - - Flores et al.2003 Up arrow = significant (P<0.01), NE= No effect (P<0.01),
  • 17. Manipulation of rumen with Ionophores • Ionophores are antibiotics includes: monensin, lasalocid, tetronasin, salinomycin, lysocellin, narasin, nigericin, laidlomycin and valynomycin • Since the mid-1970’s they have been extensively used to manipulate rumen fermentation • ionophores are approved in several countries including Australia, Argentina, New Zealand, and South Africa and recently USA • Monensin (trade name Rumensin) widely used ionophores
  • 18. Cont’d… • Effect of ionophores on rumen end products • decrease in hydrogen, a precursor of methane, • Favors propionate production • Ionophores inhibit gram-positive bacteria • major benefit of feeding ionophores to dairy cattle is the potential increase in production of propionate and the associated decrease in production of methane, which conserves energy •
  • 19. Safety (residual effect) • Several researchers indicated that meat and milk produced from animals fed monensin is safe for human consumption • Likewise, monensin is biodegradable in manure and soil, and is not toxic for crop plants
  • 20. The effects of ionophores on fermentation end products and total ciliate protozoal populations in Angus yearling steers •CC= control •CM=33 mg/kg Monensin • CM/L= Biweekly rotation of monensin (33 mg/kg) and lasalocid (36 mg/kg) (Source: Guan et al. 2005)
  • 21. Manipulation of ruminal fermentation by plant extracts • Plant extracts have been used for centuries for various purposes (as traditional medicine and food preservatives, among others • The antimicrobial activity of plant extracts - secondary plant metabolites (saponins, tannins, and essential oils) • selective inhibitors of methanogens
  • 22. Cont’d… • Plants exhibiting anti-methanogenic activity include: Equisetum arvense, Lotus corniculatus, Rheum palmatum, Salvia officinalis, Sapindus saponaria, Uncaria gambir and Yucca schidigera • Major commercial source of saponins is Yucca schidiger • Defaunating agent and selective inhibitor- gram positive bacteria
  • 23. Effect of Yucca Schidigera extract ( Saponins) in vitro ruminal characteristics and gas production Yucca schidigera extract (ml/l) SEM P value 0 2 4 6 pH 6.76 6.79 6.74 6.72 0.02 NS NH3-N (mg/l) 28.60 21.15 18.19 14.86 1.61 *** TVFA(mM) 66.45 64.22 67.97 69.24 1.57 NS VFA (mM) Acetate (A) 47.71 44.12 45.50 46.26 1.05 NS Propionate (P) 12.37 14.20 17.07 19.02 0.49 *** Butyrate 5.69 5.54 4.92 3.64 0.19 *** Others 0.67 0.35 0.49 0.32 0.11 NS A:P ratio 4.42 3.88 3.61 3.52 0.06 *** Protozoa (x104/ml) 13.16 9.38 5.90 5.80 1.07 *** CH4 (ml/min) 0.24 0.20 0.17 0.14 0.00 *** CO2 (ml/min) 1.22 1.21 1.39 1.56 0.06 *** ( Source: Pen, 2007)
  • 24. Conclusions • Rumen manipulation has paramount importance in improving ruminal fermentation efficiency and productivity in dairy and fattening ruminants in the tropics where the major sources of the feed is high in fibre. • Rumensin from ionophores and Malic acid from organic acid are the widely used rumen modifiers in both dairy and fattening ruminant animals
  • 25. Cont’d… • Several literatures reported that use of one type of manipulating technique for prolonged time is not effective because of adaptation by the rumen microbes, hence, alternative or rotational use rather than one alone may be prolonged its effect in improving the performance of the animal • The studies conducted with plant extracts are limited to in vitro screening for the desired characteristics and only a few of them have been tested in in vivo experiments
  • 26. Cont’d… • it is desirable to standardize in vivo dose of these plant extracts which are effective in inhibiting methane emission with minimum adverse effects on fermentation of feed so that these can be practically exploited for economic and ‘ecologically friendly’ livestock production
  • 27. Thank You for your kind attention

Notas do Editor

  1. Eminar on