RUMEN ECOSYSTEM AND CLASSIFICATION OF RUMEN MICROBES.pptx

RUMEN MICROBIAL ECOSYSTEM AND CLASSIFICATION OF RUMEN MICROBES

1. TATHAGAT P. KHOBRAGADE MVSc Scholar (Ani. Nutrition) ANN - 612 RUMEN BIOTECHNOLOGY 1+0 RUMEN ECOSYSTEM M.V.Sc DEPARTMENT OF ANIMAL NUTRITION COLLEGE OF VETERINARY SCIENCE AND ANIMAL HUSBANDARY, KAMDHENU UNIVERSITY, JUNAGADH

2. Learning Objectives:- INRODUCTION TO RUMEN ECOLOGY CLASSIFICATION OF RUMEN MICROBES RUMEN BACTERIA AND THEIR TYPES RUMEN PROTOZOA RUMEN FUNGI RUMEN BACTERIOPHAGES

3. INTRODUCTION TO RUMEN :- Ruminant animal has complex stomach that is divided into four chambers i.e., Rumen, Reticulum, Omasum And Abomasum . Ex. Cow, Buffalo, Goat. The Rumen (also called as “ Paunch ”) is the largest compartment of ruminal stomach and is the first chamber in the alimentary canal of ruminant animals. The Capacity of an Adult Animal Rumen is about 150 Liters . Rumen is one of the Most Dense Microbial Habitats in the world.

4. The foods of ruminants – forages and fibrous roughages – consist mainly of Polysaccharides Such As Cellulose , Which Cannot Be Broken Down By Mammalian Digestive Enzymes . Ruminants have therefore evolved a special system of digestion that involves microbial fermentation of food before its exposure to their own digestive enzymes Microscopic organisms called Rumen Microbes break down (or digest) the ingested feed by a fermentation process and this microbes comprise the Rumen Ecosystem. Rumen Microorganisms Produce Enzymes During Digestion Process.

5. a mortal enemy PHYSIOCHEMICAL PROPERTIES OF THE RUMEN:-

6. The ruminal ecosystem consists of a wide diversity of microorganisms that are in a symbiotic relationship in a strict anaerobic environment. It consist of :- • Bacteria (10¹⁰ to 10¹¹ organisms/ml), • Bacteriophage (10⁸ to 10⁹ organisms/ml), • Protozoa (10⁵ to 10⁶ organisms/ml), • Fungi (10³ to 10⁴ organisms/ml) Anaerobic Ciliate Protozoa and Anaerobic Bacteria and Anaerobic Fungi are the major microbes in rumen. The anaerobiosis inside the rumen is one of the major constraints in the rumen eco-system, which helps in conserving the energy ultimately to be used by the host animal. CLASSIFICATION OF RUMEN MICROBES :-

7. • The main advantage of rumen microbes is their ability to use plant polysaccharides and non-protein nitrogen in a diet, which cannot be digested and utilized by the host. • In ruminants, microorganism breakdown the higher carbohydrates, cellulose, pentosans and starch (protein as well) to monosaccharides and then fermented to VFA and methane. • Microbial mass synthesized in the rumen provides about 20% of the nutrients absorbed by the host animal. • Microbial fermentation of carbohydrates and protein yield volatile and short chain fatty acids which provide 60-80% of total metabolizable energy . • Microorganisms synthesize essential nutrients such as B-vitamins, amino acids. IMPORTANCE OF THE RUMEN ECOSYSTEM: -

8. The rumen contains a variety of bacterial genera, which constitute most microorganisms that live in anaerobic environments . RUMEN BACTERIA :-

9. Cellulose Degrading Bacteria Microorganisms Fermentation products Fibrobacter succinogenes Succinate, Acetate, Formate Butyrivibrio fibrisolvens Acetate, Formate, Lactate, Butyrate, H2, CO2 Ruminococcus albus Acetate, Formate, H2, CO2 Clostridium lochheadii Acetate, Formate, Butyrate, H2, CO2 Cellulose Degrading Bacteria produce extracellular cellulase enzymes to break β (1-4)- glycosidic bonds of the cellulose and produce monosaccharide glucose The ruminant diet is mainly plant based and cellulose is the main component of the cell wall of these plants. So, cellulolytic ruminal microorganisms play an important role in animal nourishment. Fibrobacter succinogenes

10. Starch is an important component of the diet of cattle and high milk producing cows which are fed with concentrates containing major proportions of grains . Starch is an easily fermentable energy source for ruminants . When starch undergoes ruminal fermentation in Rumen, Amylolytic Bacteria produce Maltase enzyme that convert Starch into Disaccharide Maltose and further into monosaccharide glucose. Amylolytic Bacteria Microorganisms Fermentation products Bacteriodes ruminicola Formate, Acetate,Succinate Ruminobacter amylophilus Formate, Acetate, Succinate Selenomonas ruminantium Acetate, Propionate, Lactate Succinomonas amylolítica Acetate, Propionate, Succinate Ruminobacter amylophilus

11. When diet containing high amount of concentrate is being fed, Lactate is produced as an intermediary product of ruminal fermentation, which is then metabolized to VFAs . Lactate utilizing bacteria have an important role in the rumen fermentation, mainly in those ruminants that are being fed with high grains in the diet. These bacteria metabolize lactic acid and prevent its accumulation, which helps to keep the pH in the proper range. Lactate Utilizing Bacteria Microorganisms Fermentation products Selenomonas lactilytica Acetate, Succinate Megasphaera elsdenii Acetate, Propionate, Butyrate, Valerate, H2, CO2 Selenomonas lactilytica

12. Pectin is fermented by both bacteria and protozoa. The major bacteria that perform this function are Butyrivibrio fibrisolvens, Prevotella ruminicola, Bacteroides ruminicola and Lachnospira multiparus . These ruminal bacteria produce and release pectinolytic enzymes primarily pectin lyases into the ruminal environment that hydrolyze the pectin in to simple form. Pectin Degrading Bacteria Proteolytic Bacteria In ruminants, proteins serve as a source of nitrogen for rumen microbes to make their own microbial protein. Microbes can use non protein nitrogenous substances also such as urea for microbial protein synthesis. Urea is 100% degradable in the Rumen by Microbial Urease and can be toxic at higher levels.

13. Microorganisms Fermentation products Bacteroides amylophilus NH3, Amino acid, VFAs Bacteroides ruminicola NH3, Amino acid, VFAs Butyrivibrio fibrisolvens NH3, Amino acid, VFAs In ruminant animals, the lipid content of the diet is low and comes from different sources such as grass, leaves, oil seeds, or cereal grains. The major types of lipids in the diet are triglycerides, phospholipids and galactolipids. When dietary lipids enter the rumen, hydrolysis of dietary lipids is brought by microbial lipases , which releases glycerol and free fatty acids from the lipid backbone. Glycerol is readily metabolized by the rumen bacteria to form propionic acid. Ex. Anaerovibrio lipolytica produces Acetate, Propionate Lipolytic Bacteria

14. Ruminal Archaea Or Methanogens Methane (CH4) is one of the end product of ruminal fermentation and is considered as a loss of total energy consumed by ruminants, representing 6-10% of total energy, which contributes to the greenhouse effect. The Methanogens belong to the Domain Archaea and the Phylum Euryarchaeota. Methane is generated by methanogenic bacteria utilizing the carbon dioxide and hydrogen. Methanogenesis is the main sink of H+ removal. CO2 is used by the methanogens to produce CH4. These microorganisms can also degrade substrates containing methyl or acetyl groups, such as methanol and acetate that act as electron acceptors. Microorganisms Fermentation products Methanobrevibacter ruminantium Methane Methanomicrobium mobile Methane CO 2 + 4H 2 CH 4 + 2H 2 O Methanobrevibacter ruminantium

15. RUMEN PROTOZOA:- Protozoa are group of single-celled eukaryotes , either free- living parasitic, that feed on organic matter such as other microorganisms or organic debris. Rumen Protozoal population is up to 10 6 per ml. All the protozoa (2% of the weight of rumen contents) are strictly anaerobic . Flagellates are less; cilates are predominating . The digestion of cellulose by the larger entodiniomorphid protozoa Rumen protozoa are responsible for 30-40% of total rumen microbial fibre digestion. Morphological studies have identified more than 250 species of ciliates living in the various ruminants. Isotricha intestinalis

16. The rumen ciliates are of two types ,which have been studied inside the rumen :- Holotrichs ( cilia present on all over the body ) Holotrichs use – soluble carbohydrates Isotricha intestinalis , Dasytricha ruminantium , Charon eguii Blepharoprosthium pierum . Oligotrichs (Entodiniomorphs – cilia present on one side of body ) Entodiniomorphs (oligotriphs ) - use particulate material like starch Entodinium busa ,Diplodinium cristagalli , Diploplastron affine Ostracodinium gracile , Epidinium caudate , Ophryoscolex purkynei Eudiplodinium maggaii , Metadinium medium . They engulf bacteria and feed particles and digest carbohydrates, proteins and fats . They utilize large amounts of starch at one time and can store it in their bodies.

17. The rumen ciliates also have proteolytic activity and produce ammonia and amino acids as end products. Their nitrogen metabolism is based largely on the digestion of engulfed bacteria , although all rumen ciliates contain enzymes capable of digesting plant proteins. Proteolytic protozoa Fermentation products Entodinium caudatum Ciliates Ammonia, VFA Eudiplodinium medium Ciliates Ammonia, VFA

18. The rumen fungi are anaerobic, zoospore forming and very sensitive to oxygen. The fungi are established within first 2 weeks of age. Dominant role in the degradation of lignocellulosic components of the feed particles. The organisms have been shown to be important for initiation and continuation of rumen fermentation. Rumen fungi have amylolytic and proteolytic activities. Anaerobic fungi are among the most active organisms in fiber digestion . Rumen Fungi belongs to the Class Neocallimastigomycetes , consisting of six previously recognized genera (Anaeromyces, Caecomyces, Cyllamyces, Neocallimastix, Orpinomyces and Piromyces). RUMINAL FUNGI:-

19. Cellulolytic fungi :- Ruminal fungi are able to produce enzymes that hydrolyze cellulose and xylans . Neocallimastix frontalis, Piromyces joyonii, and Orpinomyces communis , can more efficiently digest structural polysaccharides than cellulolytic bacterial species in monoculture. The fungal activity helps the ruminal digestion of the plant cell wall. Genus Species Fermentation Products Neocallimastix N. Frontalis N. patriciarum N. variabilis Lactate, formate, acetate, succinate, ethanol, cellobiose, oligosaccharides glucose Caecomyces C. Communis , C. equi C. sympodialis Lactate, formate, acetate, succinate Orpinomyces O. bovis O. joyonii O. intercalaris Lactate, formate, acetate, succinate, ethanol, cellobiose, oligosaccharides glucose

20. BACTERIOPHAGE:- Bacteriophages are one of the most important component of the rumen present typically at 10 8 -10 9 organisms/ml of rumen fluid. These are specific for different bacteria present in the rumen. These are also considered to be obligate pathogens for the bacteria , as bacteriophages are capable of lysing/engulfing the bacteria. By lysing the bacterial cells, bacterial protein is easily made available to the animals as a source of amino acids. The specificity of the bacteriophages for a particular rumen bacterium may be exploited for removal or killing by lysis of unwanted rumen bacteria from the ecosystem like Streptococcus bovis and Methanogens. These viruses belonged to 3 distinct families of tailed phages ; the Myoviridae, Siphoviridae, and Podoviridae, are the predators of ruminal bacteria of the genera Bacteroides, Ruminococcus, and Streptococcus.

21. Siphoviridae such as Methanobacterium phage Ψ M1, Methanobacterium phage Ψ M10, Methanobacterium phage Ψ M100, Methanobacterium phage Ψ M2 infect Methanogens like Methanobrevibacter ruminantium , Methanomicrobium mobile. are important Rumen Bacteriophages. Siphoviridae (A–E),Myoviridae (F–I), Podoviridae (J–O) and other virus-like-particles including potentially non-tailed phages (P–S).

22. Choudhury Prasanta & A.Z.M., Salem, & Jena, Rajashree & Kumar, Sanjeev & Singh, Rameshwar & Puniya, Anil. (2015). Rumen Microbiology: An Overview. Rumen Microbiology: From Evolution to Revolution. 1. 10.1007/978-81-322-2401-3_1. D.V. Reddy : Applied Nutrition (3rd ed.), CBS Publishers & Distributors Pvt Ltd., 2015. Pages 101- 112. Mackie RI, McSweeney CS, Aminov RI (1999) Rumen. In: Encyclopedia of life sciences. Nature Publishing Company, London Shreya Patel, Padma Ambalam. Role of Rumen Protozoa: Metabolic and Fibrolytic. Adv Biotech & Micro. 2018; 10(4): 555793. DOI: 10.19080/AIBM.2018.10.555793 William H. Hoover, Tammy K. Miller, Rumen Digestive Physiology and Microbial Ecology, Veterinary Clinics of North America: Food Animal Practice, Volume 7, Issue 2, 1991 Pages 311-325, ISSN 0749-0720, https://doi.org/10.1016/S0749-0720(15)30801-X. References

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