Clay Mineral Minerals as a Strategy for Biomolecule Incorporation: Amino Acids Approach

The potential use of amino acids by ruminal microorganisms converting them into microbial protein for ruminants makes it challenging to supplement these nutrients in an accessible form in animals’ diets. Several strategies to protect amino acids from ruminal degradation were reported, producing amino acids available for the protein used in the intestine called “bypass.” The intercalation of biomolecules in clay mineral minerals has gained notoriety due to its ability to support, protect, transport, physicochemical properties and non-toxicity. This study aimed to investigate the incorporation of L-lysine (Lys), L-methionine (Met), and L-tryptophan (Trp) amino acids in the clay minerals sepiolite (Sep) and Veegum® (Veg) using the adsorption method. The characterization techniques of X-ray diffraction and infrared spectroscopy indicated the presence of biomolecules in the inorganic matrices. Elemental and thermal analyzes monitored the percentages of incorporated amino acids. They showed better incorporation capacities for Veg, such as Met-Veg < Lys-Veg < Trp-Veg and Lys-Sep < Met-Sep < Trp-Sep for sepiolite, except for the incorporation of Met. Matrices provide a promising alternative for planning the administration of biomolecules, using essential amino acids as models, and may offer an alternative to improve functional diet strategies.

Bioprospecting of cow's ruminal microbiota from a slaughterhouse in Ambarawa, Central Java, Indonesia

Murwani R, Sibero MT, Silitonga PRA, Ambariyanto A. 2021. Bioprospecting of cow's ruminal microbiota from a slaughterhouse in Ambarawa, Central Java, Indonesia. Biodiversitas 22: 5030-5038. Ruminal microorganisms play essential roles in maintaining ruminant health. However, most studies focused only on ruminal lactic acid bacteria (LAB), although other ruminal microorganisms might have biological properties for biotechnological purposes. Therefore, the current study aimed to isolate ruminal bacteria (LAB and non-LAB) and fungi from ruminal material and conducted a bioprospecting study to understand their ability to produce antibacterial compounds and polysaccharide-degrading enzymes. The ruminal bacteria were isolated on MRS and ISP4 agar, while PDA was used to isolate the different fungi. The antibacterial property was tested against multidrug-resistant Escherichia coli and Salmonella enterica ser. Typhi. The ability to produce agarase, alginate-lyase, and carrageenase was screened. Prospective isolates were identified using DNA barcoding approach. Twelve bacteria were isolated using MRS agar, six from ISP4 agar, and four fungi from PDA. Among twelve bacteria from MRS agar, eleven were considered LAB, which consisted of Lactobacillus plantarum and Pediococcus acidilactici. Several classes of bacteria such as actinobacteria, firmicutes, ?-proteobacteria, and ?-proteobacteria were isolated during this study. In addition, three fungal classes, namely Saccharomycetes, Eurotiomycetes, and Mucoromycetes were also isolated. All bacteria from MRS agar were suggested to have potential compounds with antimicrobial activity, while all ruminal fungi exhibited potential sources of polysaccharide-degrading enzymes.

Calorimetry, physicochemical characteristics and nitrogen release from extruded urea

Our hypothesis was that extrusion of urea associated with corn may reduce N solubilization and increase the nutritional quality of this food for ruminants. We aimed to physically and chemically characterize a corn and urea mixture before and after the extrusion process. It was evaluated morphological differences by scanning electron microscopy, nitrogen solubilization, and compound mass loss by thermogravimetry. In scanning electron microscopy, extruded urea showed agglomerated and defined structures, with changes in the morphology of starch granules and urea crystals, differing from the arrangement of the corn and urea mixture. The extruded urea maintained a constant nitrogen release pattern for up to 360 min. In thermogravimetry, extruded urea presented a higher temperature to initiate mass loss, that is, the disappearance of the material with increasing temperature, but the mass loss was lower when compared to the first event of the corn and urea mixture. In conclusion the process of extrusion of urea with corn modifies the original structures of these ingredients and controls the release of nitrogen from the urea, maintaining in its formation an energy source optimizing the use of nitrogen by ruminal bacteria, because the more synchronized the release of starch (energy) and nitrogen, the better the use by ruminal microorganisms.

Dandelion (Taraxacum mongolicum Hand.-Mazz.) Supplementation-Enhanced Rumen Fermentation through the Interaction between Ruminal Microbiome and Metabolome

This study investigated the effects of dandelion on the ruminal metabolome and microbiome in lactating dairy cows. A total of 12 mid-lactation dairy cows were selected and randomly classified into two groups, supplementing dandelion with 0 (CON) and 200 g/d per cow (DAN) above basal diet, respectively. Rumen fluid samples were collected in the last week of the trial for microbiome and metabolome analysis. The results showed that supplementation of DAN increased the concentrations of ammonia nitrogen, acetate, and butyrate significantly. The rumen bacterial community was significantly changed in the DAN group, with Bacterioidetes, Firmicutes, and Proteobacteria being the main ruminal bacterial phyla. The abundance of Ruminococcaceae_NK4A214_group, UCG_005, and Christensenellaceae_R_7_group were relatively higher, whereas that of Erysipelotrichaceae_UCG_002 and Dialister were lower in the DAN than those in the CON. Metabolomics analysis showed that the content of d-glucose, serotonin, ribulose-5-phosphate, and d-glycerate were higher in the DAN group. These metabolites were enriched in the starch and sucrose metabolism, pentose phosphate pathway, tryptophan metabolism, and glycerolipid metabolism. The ribulose-5-phosphate and d-glycerate were correlated with Ruminococcaceae_NK4A214_group, UCG_005, and Christensenellaceae_R-7_group positively. This study demonstrated that the supplementation of dandelion impacts the ruminal microorganisms and metabolites in a way that rumen fermentation was enhanced in lactating dairy cows.

139 Ruminal microbiota mineral requirements to optimize performance on different diets

This presentation will discuss mineral requirements of ruminal microorganisms, and the effect of trace mineral source on ruminal fermentation. Sulfur and phosphorus are required in relatively large amounts by ruminal microorganisms, and dietary deficiencies of these minerals have been related to impaired ruminal fermentation. A number of trace minerals are required in low concentrations by ruminal microorganisms. With the except of cobalt (Co) minimal trace mineral requirements of the host ruminant appear to be considerably greater than that needed for rumen microbial requirements. It is well known that certain bacteria can synthesize vitamin B12 from inorganic Co. Some bacteria require vitamin B12 as a growth factor, and adequate dietary Co is needed to allow sufficient ruminal B12 synthesis to meet their requirement. Vitamin B12 is needed as a cofactor for ruminal microorganisms to convert succinate to propionate. Dietary Co deficiency results in decreased ruminal propionate in ruminants fed high concentrate diets, and decreased fiber digestion in ruminants fed high fiber diets. Attempts have been made to use high concentrations of certain trace minerals to favorably manipulate ruminal fermentation. For example, attempts have been made to increase rumen protein bypass by feeding high dietary zinc (Zn). However, studies have indicated that high concentrations of copper (Cu), Zn, and iron reduce cellulose digestion in vitro. Recent studies have indicated lower fiber digestibility in cattle supplemented with sulfate sources of Cu, Zn, and manganese compared with those fed similar concentrations from hydroxy or certain organic sources. Additional research is needed to elucidate the mechanism(s) whereby trace mineral sources affect fiber digestibility differently.

Ruminal epithelium: a checkpoint for cattle health

The reticulorumen, as the main fermentation site of ruminants, delivers energy in the form of short-chain fatty acids (SCFA) for both the animal as well as the ruminal wall. By absorbing these SCFA, the ruminal epithelium plays a major role in the maintenance of intraruminal and intraepithelial acid–base homoeostasis as well as the balance of osmolarity. It takes up SCFA via several pathways which additionally lead to either a reduction of protons in the ruminal lumen or the secretion of bicarbonate, ultimately buffering the ruminal content effectively. Nutrition of the epithelium itself is achieved by catabolism of the SCFA, especially butyrate. Catabolism of SCFA also helps to maintain a concentration gradient across the epithelium to ensure efficient SCFA uptake and stability of the epithelial osmolarity. Furthermore, the ruminal epithelium forms a tight barrier against pathogens, endotoxins or biogenic amines, which may emerge from ruminal microorganisms and feed. Under physiological conditions, it reduces toxin uptake to a minimum. Moreover, the epithelium seems to have the ability to degrade biogenic amines like histamine. Nonetheless, in high performance production animals like dairy cattle, the reticulorumen is confronted with large amounts of rapidly fermentable carbohydrates. This may push the epithelium to its limits, even though it possesses a great capacity to adapt to varying feeding conditions. If the epithelial limit is exceeded, increasing amounts of SCFA lead to an acidotic imbalance that provokes epithelial damage and thereby elevates the entrance of pathogens and other potentially harmful substances into the animal's body. Hence, the ruminal epithelium lays the foundation for the animal's health, and in order to ensure longevity and high performance of ruminant farm animals, it should never be overburdened.

Identifying Plants that Reduce Methane Production Using an In Vitro System—Helping the Challenge to Become C Neutral

The Australian red meat industry has set a goal to be carbon neutral by 2030. Reaching this goal will be a challenge and will involve targeting ways to increase carbon in the landscape, improve efficiency of production and reduce methane emissions from ruminants. There are a number of different options the industry can pursue to try and achieve its goal, including changing grazing management practices and land-use to changing the animal, what it eats and the microbial ecology in their rumen. No single one of these options will enable the red meat industry to become carbon neutral by 2030, it will take a combination of all of them to help meet the challenge. We have been using an in vitro batch fermentation system and a Rusitec system as a quick, relatively inexpensive, way to screen; plants that already exist in our grazing systems, novel plants, plant extracts and organic waste products from the horticultural industry, for their potential to improve the efficiency of fermentation and reduce methane production in the rumen. We have also used these systems to provide an initial clue about the mechanism of action at the level of the ruminal microorganisms. We have identified variation in these traits amongst the plants, plant extracts and horticultural waste products we have tested that could help develop systems that reduce the environmental footprint of ruminants in tropical production systems in Australia and in other parts of the world.

Effects of Urtica cannabina to Leymus chinensis Ratios on Ruminal Microorganisms and Fiber Degradation In Vitro

The study was conducted in vitro to investigate the effects of different ratios of Urtica cannabina and Leymus chinensis on fiber microstructure and digestibility in ruminal fluid. The experiment was divided into five groups based on the U. cannabina/L. chinensis ratios: A (0:100), B (30:70), C (50:50), D (70:30), and E (100:0). The culture medium was collected at 0, 1, 3, 6, 12, and 24 h. The results showed that: (1) in vitro crude protein degradability (IVCPD) was higher in group A, whereas in vitro neutral detergent fiber degradability (IVNDFD) was higher in group C (p < 0.05); (2) protozoa count was increased from 1 h to 3 h and decreased afterwards, with significant differences observed in several genera (p < 0.05); (3) microbial crude protein (MCP) contents at 1, 3, 6, and 24 h were higher in groups A and C (p < 0.05); (4) the basic tissue of U. cannabina was gradually degraded. At 24h, the secondary xylem vessel structure was observed in groups B and C, but not in groups D and E. In summary, there was higher neutral detergent fiber (NDF) digestibility, higher rumen MCP contents, and lower protozoa count, showing the significance of the 50:50 ratio for microbial growth and fiber digestibility.

Microbial Patterns in Rumen Are Associated with Gain of Weight in Beef Cattle

Ruminal microorganisms play a pivotal role in cattle nutrition. The discovery of the main microbes responsible for enhancing the gain of weight in beef cattle might be used in therapeutic approaches to increase animal performance and cause less environmental damages. Here, we examined differences in bacterial and fungal composition of rumen samples of Braford heifers raised in a natural grassland from Pampa Biome in Brazil. We aimed to detect microbial patterns in the rumen that could be correlated with the gain of weight. 16S and ITS1 genes were amplified from ruminal samples and sequenced to identify the closest microbial relatives within the microbial communities. A predictive model based on microbes responsible for the gain of weight was build and further tested using the entire dataset. The model detected a set of microorganisms associated with animals in the high gain of weight group, including the bacterial taxa RFN20, Prevotella, Anaeroplasma and RF16 and the fungal taxa Aureobasidium, Cryptococcus, Sarocladium, Pleosporales and Tremellales. Most of these organisms have been correlated to the production of substances that improve the ruminal digestion process. These findings provide new insights about cattle nutrition and suggest the use of these microbes to improve beef cattle breeding.