• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.12
• /
• pp.1725-1731
• /
• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.6
• /
• pp.988-1001
• /
• 1999

The rumen ecosystem is increasingly being recognized as a promising source of superior polysaccharide-degrading enzymes. They contain a wide array of novel enzymes at the levels of specific activities of 1,184, 1,069, 119, 390, 327 and $946{\mu}mol$ Reducing sugar release/min/mg protein for endoglucanase, xylanase, polygalactouronase, amylase, glucanase and arabinase, respectively. These enzymes are mainly located in the surface of rumen microbes. However, glycoside-degrading enzymes (e.g. glucosidase, fucosidase, xylosidase and arabinofuranosidase, etc.) are mainly located in the rumen fluid, when detected enzyme activities according to the ruminal compartments (e.g. enzymes in whole rumen contents, feed-associated enzymes, microbial cell-associated enzymes, and enzymes in the rumen fluid). Ruminal fungi are the primary contributors to high production of novel enzymes; the bacteria and protozoa also have important functions, but less central roles. The enzyme activities of bacteria, protozoa and fungi were detected 32.26, 19.21 and 47.60 mol glucose release/min/mL mediem for cellulose; 42.56, 14.96 and 64.93 mmol xylose release/min/mL medium after 48h incubation, respectively. The polysachharide-degrading enzyme activity of ruminal anaerobic fungi (e.g. Neocallimastix patriciarum and Piromyces communis, etc.) was much higher approximately 3~6 times than that of aerobic fungi (e.g. Tricoderma reesei, T. viridae and Aspergillus oryzae, etc.) used widely in industrial process. Therefore, the rumen ecosystem could be a growing source of novel enzymes having a tremendous potential for industrial applications.

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.1
• /
• pp.84-92
• /
• 1999

The rumen microbial ecosystem is coming to be recognized as a rich alternative source of genes for industrially useful enzymes. Recent advances in biotechnology are enabling development of novel strategies for effective delivery and enhancement of these gene products. One particularly promising avenue for industrial application of rumen enzymes is as feed supplements for nonruminant and ruminant animal diets. Increasing competition in the livestock industry has forced producers to cut costs by adopting new technologies aimed at increasing production efficiency. Cellulases, xylanases, ${\beta}$-glucanases, pectinases, and phytases have been shown to increase the efficiency of feedstuff utilization (e.g., degradation of cellulose, xylan and ${\beta}$-glucan) and to decrease pollutants (e.g., phytic acid). These enzymes enhance the availability of feed components to the animal and eliminate some of their naturally occurring antinutritional effects. In the past, the cost and inconvenience of enzyme production and delivery has hampered widespread application of this promising technology. Over the last decade, however, advances in recombinant DNA technology have significantly improved microbial production systems. Novel strategies for delivery and enhancement of genes and gene products from the rumen include expression of seed proteins, oleosin proteins in canola and transgenic animals secreting digestive enzymes from the pancreas. Thus, the biotechnological framework is in place to achieve substantial improvements in animal production through enzyme supplementation. On the other hand, the rumen ecosystem provides ongoing enrichment and natural selection of microbes adapted to specific conditions, and represents a virtually untapped resource of novel products such as enzymes, detoxificants and antibiotics.

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.1
• /
• pp.77-83
• /
• 1999

Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

• Asian-Australasian Journal of Animal Sciences
• /
• v.2 no.3
• /
• pp.400-401
• /
• 1989

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.3
• /
• pp.389-393
• /
• 2003

Present study investigate the effect of enzyme supplementation, methods (applied to rumen or enzyme treated diet) compared with no enzyme diet, on rumen fermentation and apparent nutrient digestibility in a $3{\times}3$ Latin square design with three rumen cannulated Korean Native goats. In situ rumen degradation kinetics was studied in three rumen cannulated Holstein steers. Three diets were, no enzyme, 1% enzyme in rumen and 1% enzyme in diet. The enzyme was sprayed onto forage, and the forage: concentrate ratio was 30:70. Degradation kinetics was studied with three enzyme levels (0, 1 and 2%, w/w) and four pre-treatment times (0, 1, 12 and 24 h). Results suggested that enzyme application method did not affect rumen fermentation, ruminal enzyme activity and total tract apparent digestibility. Nutrient degradation rate and effective degradability of DM, NDF and ADF increased with increasing enzyme level and pre-treatment times. Degradation of nutrients was affected by enzymes levels or pre-treatment times. Therefore, it is probable that the improved degradation may be due to the supplemented exogenous hydrolytic enzymes under a certain condition.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.3
• /
• pp.374-379
• /
• 2003

Four rumen fistulated Murrah buffaloes were used to study the effect of four diets differing in roughage to concentrate ratio on rumen biochemical changes, microbial enzyme profile and in sacco degradability of feed in a $4{\times}4$ Latin Square design. The animals were fed four diets consisting of 80:20, 70:30, 60:40 and 50:50 ratios of wheat straw and concentrate mixtures, respectively. Wheat straw and concentrate mixture were mixed with water (0.6 l/kg feed) and complete feed mixture was offered to the animals at 8:00 h and 16:00 h in two equal parts. The variation in pH of rumen liquor (difference of maximum and minimum during 0-8 h post feeding) increased with increasing level of concentrate mixture in the diet. There was no effect of diet composition on volatile fatty acids, total nitrogen and trichloro-acetic acid precipitable nitrogen in the rumen liquor, but ammonia nitrogen increased with increasing level of concentrate mixture in the ration. Major portions of all fibre degrading enzymes were present in the particulate material (PM) of the rumen contents, but protease was absent in PM fraction. The activities of micro-crystalline cellulase, acetyl esterase and protease increased with increase in the level of concentrate mixture, but the activities of other enzymes (carboxymethylcellulase, filter paper degrading activity, xylanase, $\beta$-glucosidase and $\beta$-xylosidase) were not affected. The in sacco degradability and effective degradability of feeds increased with increasing level of concentrate mixture in the ration.

• Korean Journal of Veterinary Service
• /
• v.41 no.4
• /
• pp.221-228
• /
• 2018

Rumen cannulation is used for nutritional and microbiological research, clinical diagnosis, and rumen component transfaunation. However, the cannulation procedure can affect parameters such as complete blood count findings, serum chemistry, and rumen fluid pH. The objective of this study was to evaluate the health risks related to the rumen cannulation procedure over a 1-month period. We did not identify significant differences in red blood cell numbers or morphologies between pre- and postoperative timepoints. Moreover, no inflammation or infection was detected. Despite the absence of apparent clinical signs after surgery, serum chemistry results revealed changes in blood urea nitrogen levels and the activities of liver enzymes, including aspartate transaminase, lactate dehydrogenase, and creatinine kinase, from postoperative days 1 to 14. Rumen fluid pH, as measured from samples collected via an orogastric tube, was slightly increased after a preoperative fasting period and on postoperative day 1 but decreased thereafter from postoperative day 4, indicating a minor influence of cannulation surgery on ruminal fluid pH. This is the first study to evaluate hematological parameters and rumen pH before and after rumen cannulation surgery in Hanwoo cattle. Further research is required to better elucidate the potential effects of rumen cannulation surgery on animal health.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.9
• /
• pp.1250-1260
• /
• 2010

Mycotoxins are secondary metabolites produced by fungi. These toxins pose serious health concerns to animals as well as human beings. Biodegradation of these mycotoxins has been considered as one of the best strategies to decontaminate food and feedstuffs. Biodegradation employs the application of microbes or enzymes to contaminated food and feedstuffs. Ruminants are considered to be resistant to the adverse effects of mycotoxins presumably due to the biodegrading ability of rumen microbes compared to mono-gastric animals. Therefore, rumen microbial source or microbial enzyme could be a great asset in biological detoxification of mycotoxins. Isolation and characterization of pure culture of rumen microorganisms or isolation and cloning of genes encoding mycotoxin-degrading potential would prove to have overall beneficial impact in the food and feed industry.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.1
• /
• pp.104-115
• /
• 2003

A series of experiments was carried out to determine the possibility for the non-ionic surfactant (NIS) as a feed additive for ruminant animals. The effect of the NIS on (1) the enzyme distribution in the rumen fluids of Hereford bulls, (2) the growth of pure culture of rumen bacteria and (3) rumen anaerobic fungi, (4) the ruminal fermentation characteristics of Korean native cattle (Hanwoo), and (5) the performances of Holstein dairy cows were investigated. When NIS was added to rumen fluid at the level of 0.05 and 0.1% (v/v), the total and specific activities of cell-free enzymes were significantly (p<0.01) increased, but those of cell-bound enzymes were slightly decreased, but not statistically significant. The growth rates of ruminal noncellulolytic species (Ruminobacter amylophilus, Megasphaera elsdenii, Prevotella ruminicola and Selenomonas ruminantium) were significantly (p<0.01) increased by the addition of NIS at both concentrations tested. However, the growth rate of ruminal cellulolytic bacteria (Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens and Butyrivibrio fibrisolvens) were slightly increased or not affected by the NIS. In general, NIS appears to effect Gram-negative bacteria more than Gram-positive bacteria; and non-cellulolytic bacteria more than cellulolytic bacteria. The growth rates of ruminal monocentric fungi (Neocallimastix patriciarum and Piromyces communis) and polycentric fungi (Orpinomyces joyonii and Anaeromyces mucronatus) were also significantly (p<0.01) increased by the addition of NIS at all concentrations tested. When NIS was administrated to the rumen of Hanwoo, Total VFA and ammonia-N concentrations, the microbial cell growth rate, CMCase and xylanase activities in the rumen increased with statistical difference (p<0.01), but NIS administration did not affect at the time of 0 and 9 h post-feeding. Addition of NIS to TMR resulted in increased TMR intake and increased milk production by Holstein cows and decreased body condition scores. The NEFA and corticoid concentrations in the blood were lowered by the addition of NIS. These results indicated that the addition of NIS may greatly stimulate the release of some kinds of enzymes from microbial cells, and stimulate the growth rates of a range of anaerobic ruminal microorganisms, and also stimulate the rumen fermentation characteristics and animal performances. Our data indicates potential uses of the NIS as a feed additive for ruminant animals.