10 J Contemp Med Sci | Vol. 9, No. 1, January-February 2023: 10–17 Review Ruminant Gut Microbiota: Importance, Development, and Alternative Therapeutics for Dysbiosis Nahlah Albakri1,2, Reda Amasha1, Magda M. Aly1,3* 1Department of Biology, College of Science, King Abdullaziz University, Jeddah, Saudi Arabia. 2Department of Biology, College of Science, Taibah University, Al-Medinah Al-Munawarh, Saudi Arabia. 3Botany and Microbiology Department, Faculty of Science, Kafrelsheikh University, Kafrelsheikh , Egypt. *Correspondence to: Magda M. Aly (E-mail: mmmohammad@kau.edu.sa) Abstract The microbiome is a population of microbes that colonized in mammalian gut. During the first few years of life, the gut microbiome undergoes alteration and is very diverse in adulthood, depends upon various of circumstances. Gut microbes, particularly gut flora in ruminants, are receiving more and more attention. Intestinal microbes, particularly ruminant microorganisms, have attracted an increasing amount of attention as high-throughput sequencing technology has improved and costs have decreased, whether in the fundamental research or application fields. The ruminant microbiome changes in conjunction with its host and it is influenced by inter-microbial interactions, environmental exposures, and host properties. However, any organism’s core functional microbiome is much more conventional. Unfortunately, the fragile growth ratio of the microbial culture is susceptible to incursions under illness circumstances, which may affect the abundance of various microbial species, resulting to dysbiosis. As a result, the purpose of this review is to provide a broad summary of the relevance of ruminant gut microorganisms, as well as to investigate variables that influence the microbiota and alternative therapeutics such as probiotics, prebiotics, fecal transplantation, and rumen transfiguration, all of which have been shown to be effective in addressing dysbiosis. Keywords: Gastrointestinal microbiome, alternative therapeutics, dysbiosis ISSN 2413-0516 Introduction The gut microbiota is a diverse collection of microorganisms that occur in the gastrointestinal tract of mammals. This microbial community has a host-specific composition that is continually changing and responsive to both external and endogenous changes. The importance of this “organ’s” struc- ture and function in health and illness has been emphasized by increased attention to its structure and function. The intes- tinal microbiota, a complex ecosystem, contains around 1000 species of Firmicutes, Bacteroidetes, Actinomycetes, Proteo- phylum, and Verrucomicrobia.1,2 From nutritional condition to behavior and stress response, the microbiota is closely connected to many characteristics of normal host physiology. They might also be a main or sec- ondary cause of a variety of diseases that affect both neigh- boring and distant organ systems. The presence or absence of significant species accomplished of causing particular responses, as well as the overall balance of the gut microbial community, are crucial in maintaining homeostasis or deficiency thereof in the intestinal mucosa and beyond. The mechanisms concluded which the microbiota exerts its valuable or detrimental effects are currently unknown. However, they contain both intestinal epithelial and mucosal immune cells developing signalling mol- ecules and identifying bacterial epitopes. Advances in gut microbiota modelling and analysis will help us better under- stand their role in health and illness, allowing us to tailor present and future therapeutic and prophylactic approaches.3,4 Very complicated biological materials, such as taxonomic and useful identification of microbial groups that almost fill all present ecological niches, may now be easily examined thanks to technological advances in mass spectrometry, next-generation sequencing and bioinformatics. Metaproteomic techniques give operational data on the microbiota examined, as well as struc- tural data collected via metagenomic research. Combining the major Omics technologies (metabolomics, transcriptomics, proteomics, and genomics) in living science offers very accurate data on the study object and aids in the understanding of molec- ular changes in response to externaland internalecological stimuli. The microbial communities that colonized animals’ gas- trointestinal tracts have an essential role in their metabolism as well as their physiology and health. Veterinarians, animal nutri- tionists, and microbiologists are interested in the microbiotas of cattle and ruminants.5 Between 2008 and 2014, just 20 papers were published yearly, a ratio of smaller than five per year, and there was no relevant literature before to 2008. With more than 10 papers produced per year after 2014, research entered a new era of fast expansion. The microbial diversity of the rumen or omasum and reticulum of giraffes, moose, bovine and kanga- roos, lambs, steers, Musk Deer, goats, sheep, geese, Muskoxen, yark, camel, swine, grazing primate.6 Definition of Microbiome The microbiome is a community of microorganisms that live or are present on the bodies of animals (viruses, archaea, bac- teria including fungi, protozoa, and bacteriophages) (skin, urogenital tract, oral cavity, gastrointestinal tract and respira- tory tract). The microbiome is also defined as the genetic information of microbes that inhabit in a certain environment, which includes species whose genomes are the only thing that distinguishes them.7,8 Members of the microbiome might be commensal (without harming various types that benefit from one another), symbiotic (both helpful species benefiting from one another), pathogenic, or parasitic.9 Rumen Microbiome The four components of the ruminal stomach (actual stomach) are the reticulum, rumen, omasum (pre-stomach), and (Submitted: 08 November 2022 – Revised version received: 28 November 2022 – Accepted: 03 January 2023 – Published online: 26 February 2023) 11J Contemp Med Sci | Vol. 9, No. 1, January-February 2023: 10–17 N. Albakri et al. Review Ruminant Gut Microbiota: Importance, Development, and Alternative Therapeutics for Dysbiosis abomasum.10 Because the rumen is the stomach’s most impor- tant portion, its microbial ecology enables it to use complex carbohydrates and fiber-rich diets,11 resulting in the digestion of around 70% of the cellulose consumed.12 The most abun- dant prokaryotes are bacteria, accounting for more than 95% of the ruminal microbiota.13 In the rumen, the phylum Bacte- roidetes is the most abundant, accounting for 45–57 percent 16S rRNA patterns and 90% of the Bacteroidetes population (with the genus Prevotella accounting for 45–57% 16S rRNA patterns and 90% of the Bacteroidetes population). Followed by Firmicutes accounting for 28 percent. Moreover, Dialister, Succiniclasticum, Mitsuokella, Butyrivibrio, and Ruminococcus made up over 1% of all bacterial species found in the rumen.14,15 Small Intestine Microbiome Carbohydrates and proteinabsorptionis thecore function of the small intestine,12 and its three sections (duodenum, jejunum, and ileum) have distinct roles and microbial eco- systems. According to a report, the phylum Firmicutes was more prevalent in all compartments of the cow gastrointes- tinal system except the jejunum that were dominated by Proteobacteria. In the jejunum, Ruminococcus, Acetitomac- ulum, and Lachnospiraceae were frequent, whereas Entero- bacteriaceae were prevalent in the small intestine.16 The relative abundance of the phylum Bacteroidetes reduced dramatically (0.4–1.1 percent) as compared to the rumen, while the comparative abundance of the phylum Firmicutes increased significantly (0.4–1.1%) (up to 80 percent of total relative abundance). Tenericutes (0.4–4%), Proteobacteria (0.8–5.8%), and Actinobacteria (6–13%) are examples of low-abundance taxa that have been investigated. Propioni- bacterium, Bulleidia, Mogibacterium, Lactobacillus, Mit- suokella, Ruminococcus, and Butyrivibrioare some of the other major genera found in the small intestine.14,15 Large Intestine Microbiome In the rectum, colon, and cecum, bacterial concentrations vary from 1012 to 1014 cells per millilitre.12 The large intestine is important for water digestion and absorption since it is responsible for 30 % of cellulose digestion.12,17 Various parts of the large intestine have different amounts of microbial abund- unce and diversity in the corresponding microbiota. The phylum Firmicutes have been shown to be the most abundant in the cecum, accounting for up to 81% of all phyla, with Bac- teroidetes accounting for 18–26%. Tenericutes, Actinobacteria and Spirochetes have all been discovered in the cecum. Oscillo- spira, Ruminococcus,Coprococcus, Turicibacter, Dorea, Blautia, Clostridium, and Prevotella were the most common taxa in the cecum.14 Firmicutes makes up 81% of the colon’s relative microbial abundance, with Bacteroidetes accounting for 21–33%. Furthermore, of the remaining 23 phyla, The most common bacteria were Fibrobacteria, Tenericutes, Spirochetes, Proteobacteria, and Actinobacteria. The most prevalent genera were Coprococcus, Dorea, Oscillospira, Ruminococcus, Turici- bacter, Blautia, Parabacteroides, and Prevotella.14,18 The phylum Firmicutes has invaded the rectum in a similar fashion. The major genera in the rectum were Clostridium, Prevotella, Turicibacter, Succinivibrio, Ruminococcus, Roseburia, Coproc- occus, Bacteroides, and Osillospira.19 Importance of the Rumen in Digesting Forages The rumen and associated microorganisms, such as fungi, archaea, bacteria, and protozoa, may help mammalian enzymes use substrates that aren’t easily available to them.20,21 As a result, the host ruminant has substrates that may be absorbed.22 Feeds become increasingly well-known for micro- bial colonisation after a physical breakdown during ingestion, rumination, and in the rumen.23,24 Microorganisms consume simple carbohydrates to make VFAs such acetate (for fatty acid synthesis), propionate (for glucose synthesis), and butyrate (for butyrate synthesis), which are largely used as energy sources in the ruminant body. Microbiome composition25,26 and ruminal circumstances influence the amount of various VFAs in the rumen. Intake rate, dietary forage to concentrate ratio, and diet type all impact ruminal conditions (e.g., break- down rate, molecular structure). Acetate is produced more quickly in forage-dominated diets, whereas propionate is pro- duced more easily in concentrate-based diets. Feeds heavy in starch and protein stimulate propionate synthesis, whereas hemicellulose and simple sugars raise butyrate production and cellulose increases acetate production.27 Feed must be main- tained long enough in the reticulorumen for microorganisms to adequately break down and ferment plant fibre. The supply of energy and protein in the rumen influences the efficiency and quantity of microbial protein production.28 Ruminantani- mals have a filter between the reticulum and the omasum that serves to prolong the ruminal retention time for neutral deter- gent fibre, which is an important component of forage.29 Car- bohydrates are bacteria’s primary source of energy, but they may also serve as carbon skeletons for protein synthesis when combined with amino acids, ammonia, or short peptides.30,31 During protein breakdown, peptides and amino acids are gen- erated, which may be utilized by microbes (transamination) or deaminated to form VFAs, CO2, and ammonia. 30,32 Ammonia that exceeds the ruminal wall’s microbial growth capabilities is ingested, transformed to urea, and then returned to the rumen through saliva or urine.33 Morphological and Microbial Development of the Rumen The rumen’s development and microbe colonization are a two-way interaction between the host and microbial colonies. Rumen morphological development is aided by solid feed con- sumption. VFA synthesis and absorption as fermentation end-products aid the development of ruminal papillae, allowing for their absorption and subsequent epithelial metab- olism.34,35 Butyrate is the most efficient epithelial length and function activator, followed by propionate. Roughages, on the other hand, have a physical structure that increases ruminal volume, aids muscle growth,36,37 and promotes rumen rumina- tion and saliva flow.38 The rumen’s principal enzymatic activi- ties of ruminal microbiota (proteolysis, fibrolysis, ureolysis, and amylolysis) have been reported from four to ten days of life.39,40 During the early stages of life, more than 60 glycoside hydrolase microbial genes were found in the rumen, sug- gesting a high capacity for plant carbohydrate metabolism even in the absence of regular plant cell wall consumption.41 Because 60–80% of all VFAs are absorbed across the ruminal 12 J Contemp Med Sci | Vol. 9, No. 1, January-February 2023: 10–17 Ruminant Gut Microbiota: Importance, Development, and Alternative Therapeutics for Dysbiosis Review N. Albakri et al. highly fermentable carbohydrates produces a large amount of VFA, lowering ruminal pH and causing acidosis, which may disturb physiological balance and change microbial ecology. Different kinds of roughages and the forage-to-concentrate ratio may have an impact on the ruminal metabolism and fer- mentation. Dairy cows given a mixture of Leymus chinensis hay, alfalfa hay, and corn silage produce more valerate, acetate, methylamine, and hydro cinnamate, but less glycine, glucose, isovalerate, and propionate, than dairy cows fed corn stover.56 The thickness of the stratum spinosum, stratum granu- losum, stratum basale, and stratum corneum, as well as the size of the rumen papilla, are all affected by roughage. Changes in TLR gene expression in dietary components have a compa- rable effect. Dietary compositions and types have been demon- strated to impact ruminal microbiota, rumen epithelial tissue shape, and receptors in several investigations. Ruminant farmers would benefit greatly from a good understanding of the interaction between rumen epithelial cells and rumen microbes when using dietary treatments. Age Effects on the Rumen Microbiota Another element that influences rumen microbes is the age of the host. Rumen microbes vary from newborn to 2-year-old cows.44 This might be due to dietary changes, in part (milk, colostrum, milk-supplemented rations for calves, and total mixed rations for adult cows). The host’s development is a cru- cial determinant in GIT microbiota modifications since the microbiota of cows given the same diet differs with age.41 It’s most likely due to changes in the rumen and metabolites as the animals become older. These findings imply that as the host ages, alterations in the rumen microbiome may occur. How- ever, there is a lack of knowledge on how the rumen micro- biota and the hosts interact at different ages. More research is needed to provide the knowledge needed to build innovative techniques to increasing animal production at different phases. External Environment Effects on the Rumen Microbiota The rumen microbiota and hist interaction is influenced by the external living environment, including as temperature, cli- mate, herd management, humidity, geography, and geography. The impact of the living environment on the comparability of ruminal microbiomes in cattle and bison.57 As a consequence, the microbial genomes in the GIT change as the hosts’ living environments change. The gut microorganisms and microbial activity of a ill person may vary from that of a healthy one.58 Identifying the causes of microbial diversity changes and cal- culating the host-microbiota interaction is difficult due to the complexity of the GIT environment.54 Despite the need for further study across species, nutri- tion has the greatest impact on the ruminal microbiota. Using today’s omics technology, scientists might rapidly identify the microbial makeup, host-microbe interactions, variables impacting the GIT microbiota, androles in the GIT. Future study should concentrate on the mechanisms that support the host-microbe relationship, such as dietary impacts on the GIT microbiota and epithelial cells, as well as their coordinated control.59 wall and with 75–90% of absorbed butyrate being metabolised by the ruminal epithelium, the ketogenic capacity of the rumen must develop, as a calf grows, to that of a mature rumen. After birth, microbial inoculation of the rumen began with contact with the vaginal canal, faeces, colostrum, the dam’s skin, and saliva. The presence of methanogens, fibrino- lytic bacteria, and Proteobacteria in the rumen of calves less than 20 minutes after birth has previously been investigated.42 Inoculation may happen before birth, with rapid changes in the early days of life as initial colonising aerobic or facultative anaerobic bacteria from the biotype for the later fully anaer- obic microbes.43,44 Proteobacteria were found in >90% of new- born goat sequences, showing that the rumen and epidural microbial communities are formed in the same manner.45 This might be because of their ability to scavenge oxygen from the capillary network, allowing anaerobic colonies to develop more easily. Many studies41,44,46 have shown that the prewarned rumen has the same dominating phyla as the more developed post weaned rumen, namely Bacteroidetes, Proteobacteria, and Firmicutes, with relative abundance varying with age. Fir- micutes numbers grow after weaning, however Bacteroidetes, notably Prevotella, are more reliant on solid food intake than milk elimination, reaching a stable abundance at 7 weeks and solid food consumption topping 100 g per day, respectively.46–48 This demonstrates that the earlier a calf begins to eat solid food, the sooner a mature ruminal bacterial community arises.49 looked at the effects of birth mode (natural vs. cae- sarean) and diet on the ruminal microbiome’s development, as well as random factors in early life. Others50,51 have revealed host genetic influences on the ruminal microbiome. The importance of defining ruminal function in young ruminants before, during, and after weaning was highlighted by all of these genetic and early-life effects. Factors Influencing the Microbiota-Host Interaction In reaction to environmental changes, the rumen microbiota alters its composition and function. Many scientists have studied the variables that influence the rumen microbiota throughout the past few decades. Various methods, ranging from demographic fingerprinting to high-throughput sequencing, were used. Diet, environment, and age all influ- ence the rumen microbiota. Diet Effects on the Rumen Microbiota Rumen function, which is governed by interactions between the environment andhost genes, leads to changes in rumen microbial ecology, has a significant impact on ruminant pro- duction. By modifying microbial populations and fermenta- tion activity, the type of diet may have a significant influence on rumen function.52–55 Roughage has a large influence on rumen growth, and VFA absorption is aided by gene expres- sion in rumen epithelial cells. As a result, one of the most crit- ical features of large ruminant feeding operations has been modified to enhance feed efficiency. Ruminants have acquired a digestive mechanism that can digest roughages during mil- lions of years of evolution. When ruminant animals are fed fiber-deficientor high-grain diets, the rapid breakdown of 13J Contemp Med Sci | Vol. 9, No. 1, January-February 2023: 10–17 N. Albakri et al. Review Ruminant Gut Microbiota: Importance, Development, and Alternative Therapeutics for Dysbiosis Altered Microbiota: Dysbiosis A microbiome is an interacting and changing micro-ecosystem defined by the genetic components, structures, and metabolites of unique microbiota. The microbiota may be found in a variety of environments, including among eukaryotic hosts. Many studies have identified the microbiome as a neglected niche in the maintenance of physiological functioning in eukaryotic host settings.12 The microbiome’s influence on ruminant growth and immunity has been extensively researched.13 The com- mensal microbiota contributes to animal health in a variety of ways, including helping in the digestion of indigestible plant fibre.60 It also supplies the host with sustenance and energy (vol- atile fatty acids), building components (lipids, peptides, and carbohydrates), and immune system modification (antibodies and cytokines).61 The microbiome, which competes with path- ogens for adhesion sites and nutrition, separates infections and immune cells.62,63 Furthermore, antimicrobial compounds such as hydrogen peroxide, organic acids, biosurfactants, and bacte- riocins are produced by these microbes, limiting pathogenic development.15 Dysbiosis is a breakdown or imbalance in the gastrointestinal tract’s natural microbiota. Dysbiosis may be caused by an increase or decrease in the number of commensal bacteria, the introduction of pathogenic organisms, or the development of opportunistic microorganisms. Because the microbiota is a metabolically active “organ,” dysbiosis may impact the production of essential nutrients or metabolites like short-chain fatty acids or secondary bile acids. Significant dys- biosis has been associated to acute diarrhoea (infectious, non-infectious, and hemorrhagic), chronic diarrhea (food or antibiotic response and IBD), GI motility problems, EPI, antibi- otics, and gastric acid reducers. Despite the limited evidence, dysbiosis seems to be a significant component in both acute and chronic feline and canine diarrhoea. Fatty acid, biotin, trypto- phan, ascorbate, and glycosphingolipid metabolism in the microbiota are all affected by changes in GI bacterial groups.64 Alternative Therapeutics Ways The use of Probiotics Probiotics are living associated to human and animal health that have a therapeutic effect on health when given in proper proportions.65 Probiotics’ positive benefits may be produced in a variety of ways.66 Pathogens competing for adhesion sites and nutrients may affect the host’s micro- biota.67 In addition, probiotics aid in the maintenance of intestinal homeostasis, which improves barrier function. They may create antimicrobial metabolites such as lactic acid and diacetyl, as well as antimicrobial peptides such as bacteriocins.68 By interacting directly with host cells, probi- otics have the ability to affect the immune system.69,70 The most prevalent probiotic microorganisms are food-grade bacteria from the families Bifidobacterium, Lactobacillus, and Lactobacillus. Probiotics also include microorganisms such as enterococcus and streptococcus.71 Studies injecting probiotics to treat rumen acidosis were a realistic option due to the importance of dairy cattle. According to Goto et al.,72 adding a probiotic cocktail including C. butyricum, E. faecium, and L. plantarum for seven days improved pH and lactic acid levels. Yeast has also been used in cow probiotics after many studies shown that it reduces ruminal acidity.73,74 According to Mohammed et al.,73 a diet supplemented with Saccharomyces cerevisiae decreased subacute rumen acidosis but not acute acidosis. Researchers have used probiotics as a viable alternative for preventing and managing mastitis in conjunction with global efforts to reduce antibiotic consumption.75,76 Probiotics based on LAB have been proven to increase the host immune response effectively, suggesting that they might be approved as a non-antibiotic mastitis treatment.66 The use of Prebiotics Prebiotics are natural compounds that are complexed by an animal’s superior gastrointestinal tract enzymes but can be digested, developed, and operate more efficiently by one or a small group of gut bacteria. The cumulative effect increases the host’s health.77,78 The most often used prebiotics in animals are manno-, fructose-, and trans galacto-oligosaccharides. In cattle, prebiotics have been demonstrated to reduce harmful bacteria adhesion and enhance immunological response.79 In the research, prebiotics were introduced to the Holstein Frie- sian diet, which reduced the incidence of E. coli.80 Fecal Microbiota Transplantation The therapeutic transplantation of faecal microbiota from a healthy person into an ill person is known as “faecal micro- biota transplantation,” or FMT. During the FMT procedure, all species that contribute to an entire complex ecosystem of the gastrointestinal microbiota are transplanted, including viruses, bacteria, fungi, archaea, and protozoa, as well as minute par- ticulate feedstuffs, colonocytes, and metabolites.81 FMT has a long history of usage in humans, dating back to at least the 4th century in China when it was used to treat gastroenteritis and diarrhea.82 FMT administered by colonic enemas was shown to be a successful treatment approach in four human instances of pseudomembranous enterocolitis caused by Staphylococcus aureus in an early case report published in the United States.83 FMT has been more commonly employed in hospitals and clinics in recent years as a very successful treatment option for recurrent Clostridium difficile infections that are resistant to antimicrobials.84 Although C. difficile infections are the most common ailment now treated with FMT in the modern era, many other disorders have shown a favourable response to experimental FMT treatment, includingidiopathic thrombo- cytopenic purpura, insulin sensitivity, and chronic fatigue syn- drome in patients with metabolic syndrome.85–87 The actual mechanism behind FMT’s success in the majority of disorders is unknown. Increased microbial diversity, larger numbers of beneficial microbial communities, and immune system mod- ulation are most likely to be responsible. The most common historical use of FMT in animals is in ruminants, where it is used to return microbes to the ruminal contents of cattle. It’s commonly used for digestive or meta- bolic issues, such as inappetence or ruminal hypomotility.88 Ruminant transformation has a lengthy history, dating back to the 17th century in Italy, when it was initially noted as a way for restoring correct rumination.89 For millennia, Sweden has employed regurgitated digesta or cud as a method for micro- bial transplantation to treat ruminal indigestion, with cud being described to as a “living thing”90 FMT has lately gained popularity as a treatment and prevention method in other ani- mals including household pets. 14 J Contemp Med Sci | Vol. 9, No. 1, January-February 2023: 10–17 Ruminant Gut Microbiota: Importance, Development, and Alternative Therapeutics for Dysbiosis Review N. Albakri et al. Although the FMT efficacy exact mechanism in animals, the majority of ailments is unclear, many possibilities have been offered. One of the most well-documented mechanisms of action is restoring normal flora by repopulating the gut with a full diverse population of microorganisms.91,92 Transforma- tion in ruminants, for example, is thought to be advantageous because it recolonizes the rumen with beneficial anaerobes, restoring normal fermentation function. Furthermore, increasing the variety of the microbiome enhances the host’s capacity to absorb complex carbohydrates, which aids diges- tion. FMT is believed to have a role in gastrointestinal path- ogen competitive inhibition by recolonizing normal bacteria, where beneficial microbes outcompete all the pathogens infec- tion steps.93,94 FMT has recently emerged as a viable therapy option for those afflicted with multidrug-resistant bacteria including vancomycin-resistant Enterococcus faecalis and methicillin-resistant Staphylococcus aureus.95–97 Rumen Transfaunation The rumen’s symbiotic microbiota provides nutrition and energy to the host by digesting ingesta and decomposing plant materials into different volatile fatty acids (VFAs), ammonia, and other chemicals. As a result, improved rumen microbial digesting ability may improve feed efficiency and yield. Many factors have been used in previous techniques to target the rumen microbiota. Various studies have looked at changing animal diets,1 lowering rumen pH,98 reducing rumen pro- tozoa,99 and other approaches. However, none of the aforemen- tioned procedures have consistently positive effects, suggesting that long-term ways for increasing animal performance by altering rumen microbiota have yet to be identified.100 Rumen transformation is the process of transferring rumen fluid from healthy animals to animals that have a wide variety of microbes. The transplanted rumen fluid provides energy and nutrients to the rumen microbial population.101 VFA, microbial proteins, amino acids, vitamins, enzymes and minerals are among the nutrients found in rumen fluid.102,103 Small quantities (1 L) of rumen fluid transformed, which enhances rumen function and feed intake in cows, may also be used to treat feed indigestion in cows.104 Microbial interven- tion in young calves may also be intentionally induced, affecting calf health and rumen microbiota development. Inoculating newborn animals with rumen fluid from adult animals (fresh, lyophilized, or autoclaved) improved feed effi- ciency and weight gain.105,106 Inoculating newborn lambs with mature lyophilized rumen fluid dramatically increased growth performance during and after weaning and starter meal digest- ibility, according to a recent sheep study.107 Vaccination also raised ruminal propionate concentrations and rumen amylase activity, as well as lowering the acetate/propionate ratio. The usage of mature lyophilized rumen fluid increased the population of Streptococcus ruminantium, which is linked to starch consumption.108 Similarly, in young calves fed exoge- nous rumen fluid obtained from an adult cow, the relative abundance of eight bacterial genera (Acidiphilium, Sporosar- cina, Polaribacte, Bdellovibrio, Microbacterium, Pseudodesulfo vibrio, Sporosarcina and Jeotgalibaca) belonging to four phyla Actinobacteria, Proteobacteria, Bacteroidetes, and Firmi- cutes.109 According to a recent study,110 inoculating fresh rumen fluid from adult goats promotes early rumen colonisation through the strong protozoal population, resulting in enhanced rumen absorption, feed intake and VFA production during the preweaning period. The immune systems of suckling dairy calves were improved by spray-dried rumen fluid containing 1% maltodextrin, as seen by lower interleukin-6 blood concen- trations.111 These findings suggest that providing exogenous rumen fluid to young ruminants enhances weight gain while also altering feed health, immunity, and digestibility.112 Conclusion Recently, gut microbiota has received more attention from sci- entists. The numbers of researches about intestinal micro- biota, epically in ruminant gut, are increased. Biotechnological improvement processes have positive roles in this field. Gut microbiota is a symbiotic complex ecosystem that played key roles in the host health and immune system function. These key roles are affected by both external and internal factors. Any alteration of these factors led to imbalance in the gut microbiota which is associated with dangerous diseases like dysbiosis. 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