The objective of this work was to isolate a microorganism, able to produce high lactate dehydrogenase (LDH) activity, for use as a microbial feed additive. The LDH is an important enzyme for lactate conversion in the rumen, thereby possibly overcoming lactic acidosis owing to sudden increases of cereal in the diets of ruminants. In the present study, various bacterial strains were screened from a variety of environments. Among the isolated microorganisms, strain FFy 111-1 isolated from a Korean traditional fermented vegetable food called Kimchi showed the highest enzyme activity, along with retaining strong enzyme activity even in rumen fluid in vitro. Based on morphological and biochemical characteristics as well as compositions of cellular fatty acids plus API analyses, this strain was identified as Lactobacillus sp. The optimum temperature and pH for growth were found to be 30$^{\circ}C$ and pH 6.5, respectively. A maximum cell growth of 2.2 at $A_{650}$ together with LDH activity of 2.08 U per mL was achieved after 24 h of incubation. Initial characterization of FFy 111-1 suggested that it could be a potential candidate for use as a direct-fed microbial in the ruminant animals.
A bacterial strain No. 40, which produced extracellular endoglucanase, was isolated from the rumen of Korean native goals and identified to be a genus of Actinomyces sp. The optimum conditions for endoglucanase production in PY-CMC medium were initial pH of 7.0 and 4 days of cultivation at $39^{\circ}C$. When localization of endoglucanase activity of Actinomyces sp. was determined, 68% of the enzyme activity was found in the extracellular fraction, 11% of the activity was detected in the periplasmic space and the remaining activity was in the intracellular and cell-bound fractions. The maximal endoglucanase activity was observed at pH 5.0 and it was most s table at pH 5.0. The optimum temperature of this enzyme activity was $55^{\circ}C$, but enzyme activity was gradually lost at temperature above $60^{\circ}C$. The crude enzyme was activated by addition of 10 mM cysteine and 10 mM DTT. But it was inhibited by addition of 10 mM $Cu^{{+}{+}}$ and $Fe^{{+}{+}}$. This crude enzyme could digest carboxymethylcellulose (CMC), and degrade xylan, avicel, pNPG, and pNPC to a less extent.
A 90 d study was designed to investigate the effects of rumen undegradable protein (RUP) and a mixture of Cu, Zn and Mn proteinate (CZMP) on milk yield and composition and ovarian functions during rainy months. Twenty four Holstein${\times}$ indigenous cows in their 2nd and 3rd lactation were randomly allocated to total mixed rations (TMR) containing soy bean meal (SBM) as a source of rumen degradable protein (RDP), SBM plus CZMP, and formalin treated SBM (FSBM) as a source of RUP. Maximum and minimum temperature humidity index during the experimental period were 83.6-84.7 and 75.4-76.1. There were no differences (p>0.05) in intakes of dry matter, crude protein and net energy and in contents of butterfat, lactose and minerals. Cows on TMR containing FSBM not only lost less weight (-278, -467 and -433 g/d) with more intake of RUP (0.92, 0.58 and 0.59 kg/d) but also produced more milk (19.27, 18.23 and 18.13 kg/d) and 4% fat corrected milk (18.57, 17.57 and 17.51 kg/d) with more protein (3.06, 2.81 and 2.80%), solids-not-fat (8.69, 8.38 and 8.38%) and less milk urea N (9.3, 15.4 and 15.0 mg/dl) compared with those on TMR containing SBM and SBM+CZMP, respectively (p<0.01). However, cows on TMR containing SBM and SBM+CZMP did not differ in these respects (p>0.05). Whereas incidence of cystic ovaries at 20 and 90 d pospartum was less (p<0.01) in cows on TMR containing SBM+CZMP (37.3 and 12.5%) than those on TMR containing SBM (62.5 and 25%), it was nil for cows on TMR containing FSBM. Cows in all three group differed (p<0.01) from each other for the recurrence of first observed estrus with those on TMR containing FSBM having least days (22, 36 and 47 d) compared with their counterpart on TMR containing SBM+CZMP and SBM, repectively. The results suggest that RUP is one of the limiting factors affecting milk yield and its composition and ovarian functions during early lactation of dairy cows in the tropics.
Park, Tansol;Seo, Seongwon;Shin, Teaksoon;Cho, Byung-Wook;Cho, Seongkeun;Kim, Byeongwoo;Lee, Seyoung;Ha, Jong K.;Seo, Jakyeom
Asian-Australasian Journal of Animal Sciences
/
v.31
no.4
/
pp.607-615
/
2018
Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.
Whole horse beans (Vicia faba cv. Alfred) (WHB) were pressure toasted at different temperatures of 100, 118 and $136^{\circ}C$ for 3, 7, 15 and 30 minutes in order to determine an optimal heating conditions to increase bypass starch (BPSt) as glucose source which is usually limiting nutrient in highly producing dairy cows in the Netherlands. Starch (St) Ruminal Degradative Kinetics and Fermentation Characteristics of (SRDC) of WHB were determined using in sacco technique in 4 lactating dairy cows fed 47% hay and 53% concentrate according to Dutch dairy cow requirements. Measured characteristics of St were soluble fraction (S), potentially degradable fraction (D) and rate of degradation (Kd) of insoluble but degradable fraction. Based on measured characteristics, percentage bypass starch (BPSt) was calculated according to the Dutch new feed evaluation system: the DVE/OEB system. Pressure toasting temperatures significantly affected starch gelatinization (p<0.01). Degradability of Starch in the rumen was highly reduced by pressure toasting (p<0.01). S varied from 58.2% in the raw WHB (RWHB as a control) to 19.6% in $136^{\circ}C/15min$. S was reduced rapidly with increasing time and temperature (p<0.01). D varied from 41.8% in RWHB to 80.5% in $136^{\circ}C/15min$. D fraction was enormously increased with increasing time and temperature (p<0.01). Kd varied from 4.9%h in RWHB to 3.4%/h in $136^{\circ}C/15min$. All these effects resulted in increasing %BPSt from 29.0% in RWHB to 53.1% in $136^{\circ}C/15min$. Therefore BPSt increased from 93.5 g/kg in RWHB to 173.5 g/kg in $136^{\circ}C/15min$. The effects of pressure toasting on %BPSt and BPSt seemed to be linear up to the highest values tested. Therefore no optimal pressure toasting conditions could be determined at this stage. But among 10 treatments, The treatment of $136^{\circ}C/15min$was the best with the highest BPSt content. It was concluded that pressure toasting was effective in shifting starch degradation from rumen to small intestine to increase bypass starch.
An experiment was conducted with three ruminally and intestinally cannulated non-lactating cows of Finnish Ayrshire breed, to assess the ruminal degradation characteristics of oat (Avena sativa), rye (Secale cereale) and rice (Oryza sativa) straw by the nylon bag technique, and the subsequent post-ruminal degradation of their rumen-undegraded residues by using the mobile bag technique, respectively. The straw samples were untreated or treated with aqueous $NH_3$ or with urea solution in cold or hot water. The untreated straw samples were milled or chopped, and the treated straw samples were chopped. The constant values a, b, and c were computed according to the exponential equation, where a = intercept of degradation curve at time 0, b = potentially degradable material, c = rate of degradation of band (a+b) = maximum potential degradability (asymptote). It was found that nitrogen contents of chemically treated straw were markedly increased by both $NH_3$ and urea treatments. Milling the samples attributed to a remarkable loss at 0 h incubation time as compared to chopping of the respective samples. However, chemical treatment markedly improved the b value and the subsequent (a+b) values for dry matter, organic matter, neutral-detergent fiber, and acid-detergent fiber of the samples. Furthermore, temperature of the water used in the urea solutions was considered essential, since urea in hot water rather than in cold water seemed to enhance the overall degradability. The disappearance of rumen-incubated straw residues from the mobile bags ranged from 4.5 to 9.6% for the parameters measured. On average, the OM disappearance from bags was clearly higher for the residues of urea treated straw compared to those of ammonia treated straw, but the disappearance of NDF tended, however, to be higher on the ammonia treatment.
Kim, Do Hyung;Choi, Seong Ho;Park, Sung Kwon;Lee, Sung Sill;Choi, Chang Weon
Asian-Australasian Journal of Animal Sciences
/
v.31
no.1
/
pp.80-85
/
2018
Objective: This study was conducted to investigate the effect of corn grain particle size on ruminant fermentation and blood metabolites in Holstein steers fed total mixed ration (TMR) as a basal diet to explain fundamental data of corn grain for cattle in Korea. Methods: Four ruminally cannulated Holstein steers (body weight $592{\pm}29.9kg$) fed TMR as a basal diet were housed individually in an auto temperature and humidity modulated chamber ($24^{\circ}C$ and 60% for 22 h/d). Treatments in a $4{\times}4$ Latin square design were TMR only (control), TMR with whole corn grain (WC), coarsely ground corn grain (CC), and finely ground corn grain (FC), respectively. The corn feeds substituted for 20% energy intake of TMR intake. To measure the ruminal pH, ammonia N, and volatile fatty acids (VFA), ruminal digesta was sampled through ruminal cannula at 1 h intervals after the morning feeding to determine ruminal fermentation characteristics. Blood was sampled via the jugular vein after the ruminal digesta sampling. Results: There was no difference in dry matter (DM) intake between different corn particle size because the DM intake was restricted to 1.66% of body weight. Different corn particle size did not change mean ammonia N and total VFA concentrations whereas lower (p<0.05) ruminal pH and a ratio of acetate to propionate, and higher (p<0.05) propionate concentration were noted when the steers consumed CC compared with WC and FC. Concentration of blood metabolites were not affected by different particle size of corn grain except for blood triglyceride concentration, which was significantly (p<0.05) increased by FC. Conclusion: Results indicate that feeding CC may increase feed digestion in the rumen, whereas the FC group seemed to obtain inadequate corn retention time for microbial degradation in the rumen.
Buranakarl, C.;Thammacharoen, S.;Semsirmboon, S.;Sutayatram, S.;Chanpongsang, S.;Chaiyabutr, N.;Katoh, K.
Asian-Australasian Journal of Animal Sciences
/
v.33
no.6
/
pp.921-929
/
2020
Objective: The aim of the present study was to investigate the beneficial effects of dietary supplementation with oil palm frond (leaf) (OPF) with and without oil palm meal (OPM) on nutrient intake and digestibility, ruminal fermentation and growth performance in goats. Methods: Six female crossbred goats were fed for 28 days of 3 diet treatments; 100% para-grass (T1); 50% para-grass + 50% OPF (T2), and 30% para-grass + 50% OPF + 20% OPM (T3). Body weight, rectal temperature, respiratory rate, and urine volume, food intake, dry matter intake and water intake were measured daily. Nutrient digestibility was determined from five consecutive days of last week in each diet. Ruminal fluid, urine and blood were collected at the end for determination of rumen protozoa and volatile fatty acid contents, urinary allantoin excretion, blood cell count and chemistry profiles. Results: Goats fed T2 and T3 showed higher dry matter and nutrients intakes while protein digestibility was suppressed compared with those for T1. Crude fat digestibility declined in T2 but maintained after adding the OPM (T3). High fat intake by giving OPF and OPM corresponded to a higher ruminal acetate/propionate ratio (C2/C3) and serum cholesterol level. An increased urinary allantoin/creatinine ratio was found in T2 and T3 compared with T1, implying an increased number of ruminal microbes. Conclusion: Increased dry matter intake in T2 and T3 suggested that oil palm by-products are partly useful as a replacement for para-grass in goats. Replacement with the by-products increased plasma cholesterol level, which suggested that these products are a useful energy source. Changes in rumen parameters suggested an increased microbial number and activity suitable for acetate production. However, the limited digestibility of protein implies that addition of high protein feeds may be recommended to increase body weight gain of goats.
Objective: This study was an attempt to investigate the effect of levels of fungal (Lentinus sajor-caju) treated oil palm fronds (FTOPF) on digestibility, rumen fermentation, and nitrogen balance in goats. Methods: Four 16 month old male crossbred (Thai Native${\times}$Anglo Nubian) goats with initial body weights of $33.5{\pm}1.7kg$ were randomly assigned according to a $4{\times}4$ Latin square design. Four levels of FTOPF were assigned for feed intake. The experimental treatments consisted of 0%, 33%, 67%, and 100% of oil palm fronds (OPF) being replaced by FTOPF. Results: The results revealed that total dry matter intake and nutrient intake were not influenced (p>0.05) by the inclusion of FTOPF. However, the efficiency values of the digestibility of the dry matter, organic matter, crude protein, neutral detergent fiber, acid detergent fiber, and acid detergent lignin on FTOPF were higher (p<0.05) in treatments with 33%, 67%, and 100% of FTOPF compared with 0% of FTOPF. FTOPF feeding did not change the rumen pH, temperature, and $NH_3-N$. However, the FTOPF levels did affect the total volatile fatty acid (VFA), molar proportion of acetate, propionate, butyrate, ratio of acetic (propionic acid and acetic) plus butyric (propionic acid), and production of $CH_4$. The totals of VFA and propionate was lower in goat fed with 0% of FTOPF than in those of the other groups (p<0.05). The amount of nitrogen retention based on g/d/animal or the percentage of nitrogen retained was the lowest the goat fed with 0% of FTOPF (p<0.05), whereas nitrogen intake, excretion, and absorption were not changed among treatments. Conclusion: Based on this study, FTOPF could be effectively used as an alternative roughage source in total mixed ration diets, constituting at least up to 100% of OPF.
Background: Enteric methane ($CH_4$) accounts for about 70% of total $CH_4$ emissions from the ruminant animals. Researchers are exploring ways to mitigate enteric $CH_4$ emissions from ruminants. Recently, nano zinc oxide (nZnO) has shown potential in reducing $CH_4$ and hydrogen sulfide ($H_2S$) production from the liquid manure under anaerobic storage conditions. Four different levels of nZnO and two types of feed were mixed with rumen fluid to investigate the efficacy of nZnO in mitigating gaseous production. Methods: All experiments with four replicates were conducted in batches in 250 mL glass bottles paired with the ANKOM$^{RF}$ wireless gas production monitoring system. Gas production was monitored continuously for 72 h at a constant temperature of $39{\pm}1^{\circ}C$ in a water bath. Headspace gas samples were collected using gas-tight syringes from the Tedlar bags connected to the glass bottles and analyzed for greenhouse gases ($CH_4$ and carbon dioxide-$CO_2$) and $H_2S$ concentrations. $CH_4$ and $CO_2$ gas concentrations were analyzed using an SRI-8610 Gas Chromatograph and $H_2S$ concentrations were measured using a Jerome 631X meter. At the same time, substrate (i.e. mixed rumen fluid+ NP treatment+ feed composite) samples were collected from the glass bottles at the beginning and at the end of an experiment for bacterial counts, and volatile fatty acids (VFAs) analysis. Results: Compared to the control treatment the $H_2S$ and GHGs concentration reduction after 72 h of the tested nZnO levels varied between 4.89 to 53.65%. Additionally, 0.47 to 22.21% microbial population reduction was observed from the applied nZnO treatments. Application of nZnO at a rate of $1000{\mu}g\;g^{-1}$ have exhibited the highest amount of concentration reductions for all three gases and microbial population. Conclusion: Results suggest that both 500 and $1000{\mu}g\;g^{-1}$ nZnO application levels have the potential to reduce GHG and $H_2S$ concentrations.
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