• Title/Summary/Keyword: Ruminant Nutrition

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Impact of livestock industry on climate change: Case Study in South Korea - A review

  • Sun Jin Hur;Jae Min Kim;Dong Gyun Yim;Yohan Yoon;Sang Suk Lee;Cheorun Jo
    • Animal Bioscience
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    • v.37 no.3
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    • pp.405-418
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    • 2024
  • In recent years, there has been a growing argument attributing the primary cause of global climate change to livestock industry, which has led to the perception that the livestock industry is synonymous with greenhouse gas (GHG) emissions. However, a closer examination of the global GHG emission by sector reveals that the energy sector is responsible for the majority, accounting for 76.2% of the total, while agriculture contributes 11.9%. According to data from the Food and Agriculture Organization of the United Nations (FAO), the total GHG emissions associate with the livestock supply chain amount to 14.5%. Within this, emissions from direct sources, such as enteric fermentation and livestock manure treatment, which are not part of the front and rear industries, represent only 7%. Although it is true that the increase in meat consumption driven by global population growth and rising incomes, has contributed to higher methane (CH4) emissions resulting from enteric fermentation in ruminant animals, categorizing the livestock industry as the primary source of GHG emissions oversimplifies a complex issue and disregards objective data. Therefore, it may be a misleading to solely focus on the livestock sector without addressing the significant emissions from the energy sector, which is the largest contributor to GHG emissions. The top priority should be the objective and accurate measurement of GHG emissions, followed by the development and implementation of suitable reduction policies for each industrial sector with significant GHG emissions contributions.

Evaluation of a Nutrition Model in Predicting Performance of Vietnamese Cattle

  • Parsons, David;Van, Nguyen Huu;Malau-Aduli, Aduli E.O.;Ba, Nguyen Xuan;Phung, Le Dinh;Lane, Peter A.;Ngoan, Le Duc;Tedeschi, Luis O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1237-1247
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    • 2012
  • The objective of this study was to evaluate the predictions of dry matter intake (DMI) and average daily gain (ADG) of Vietnamese Yellow (Vang) purebred and crossbred (Vang with Red Sindhi or Brahman) bulls fed under Vietnamese conditions using two levels of solution (1 and 2) of the large ruminant nutrition system (LRNS) model. Animal information and feed chemical characterization were obtained from five studies. The initial mean body weight (BW) of the animals was 186, with standard deviation ${\pm}33.2$ kg. Animals were fed ad libitum commonly available feedstuffs, including cassava powder, corn grain, Napier grass, rice straw and bran, and minerals and vitamins, for 50 to 80 d. Adequacy of the predictions was assessed with the Model Evaluation System using the root of mean square error of prediction (RMSEP), accuracy (Cb), coefficient of determination ($r^2$), and mean bias (MB). When all treatment means were used, both levels of solution predicted DMI similarly with low precision ($r^2$ of 0.389 and 0.45 for level 1 and 2, respectively) and medium accuracy (Cb of 0.827 and 0.859, respectively). The LRNS clearly over-predicted the intake of one study. When this study was removed from the comparison, the precision and accuracy considerably increased for the level 1 solution. Metabolisable protein was limiting ADG for more than 68% of the treatment averages. Both levels differed regarding precision and accuracy. While level 1 solution had the least MB compared with level 2 (0.058 and 0.159 kg/d, respectively), the precision was greater for level 2 than level 1 (0.89 and 0.70, respectively). The accuracy (Cb) was similar between level 1 and level 2 (p = 0.8997; 0.977 and 0.871, respectively). The RMSEP indicated that both levels were on average under-or over-predicted by about 190 g/d, suggesting that even though the accuracy (Cb) was greater for level 1 compared to level 2, both levels are likely to wrongly predict ADG by the same amount. Our analyses indicated that the level 1 solution can predict DMI reasonably well for this type of animal, but it was not entirely clear if animals consumed at their voluntary intake and/or if the roughness of the diet decreased DMI. A deficit of ruminally-undegradable protein and/or a lack of microbial protein may have limited the performance of these animals. Based on these evaluations, the LRNS level 1 solution may be an alternative to predict animal performance when, under specific circumstances, the fractional degradation rates of the carbohydrate and protein fractions are not known.

Meta-analysis on Methane Mitigating Properties of Saponin-rich Sources in the Rumen: Influence of Addition Levels and Plant Sources

  • Jayanegara, Anuraga;Wina, Elizabeth;Takahashi, Junichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1426-1435
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    • 2014
  • Saponins have been considered as promising natural substances for mitigating methane emissions from ruminants. However, studies reported that addition of saponin-rich sources often arrived at contrasting results, i.e. either it decreased methane or it did not. The aim of the present study was to assess ruminal methane emissions through a meta-analytical approach of integrating related studies from published papers which described various levels of different saponin-rich sources being added to ruminant feed. A database was constructed from published literature reporting the addition of saponin-rich sources at various levels and then monitoring ruminal methane emissions in vitro. Accordingly, levels of saponin-rich source additions as well as different saponin sources were specified in the database. Apart from methane, other related rumen fermentation parameters were also included in the database, i.e. organic matter digestibility, gas production, pH, ammonia concentration, short-chain fatty acid profiles and protozoal count. A total of 23 studies comprised of 89 data points met the inclusion criteria. The data obtained were subsequently subjected to a statistical meta-analysis based on mixed model methodology. Accordingly, different studies were treated as random effects whereas levels of saponin-rich source additions or different saponin sources were considered as fixed effects. Model statistics used were p-value and root mean square error. Results showed that an addition of increasing levels of a saponin-rich source decreased methane emission per unit of substrate incubated as well as per unit of total gas produced (p<0.05). There was a decrease in acetate proportion (linear pattern; p<0.001) and an increase in propionate proportion (linear pattern; p<0.001) with increasing levels of saponin. Log protozoal count decreased (p<0.05) at higher saponin levels. Comparing between different saponin-rich sources, all saponin sources, i.e. quillaja, tea and yucca saponins produced less methane per unit of total gas than that of control (p<0.05). Although numerically the order of effectiveness of saponin-rich sources in mitigating methane was yucca>tea>quillaja, statistically they did not differ each other. It can be concluded that methane mitigating properties of saponins in the rumen are level- and source-dependent.

Publication Report of the Asian-Australasian Journal of Animal Sciences over its History of 15 Years - A Review

  • Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.1
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    • pp.124-136
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    • 2002
  • As an official journal of the Asian-Australasian Association of Animal Production Societies (AAAP), the Asian-Australasian Journal of Animal Sciences (AJAS) was born in February 1987 and the first issue (Volume 1, Number 1) was published in March 1988 under the Editorship of Professor In K. Han (Korea). By the end of 2001, a total of 84 issues in 14 volumes and 1,761 papers in 11,462 pages had been published. In addition to these 14 volumes, a special issue entitled "Recent Advances in Animal Nutrition" (April, 2000) and 3 supplements entitled "Proceedings of the 9th AAAP Animal Science Congress" (July, 2000) were also published. Publication frequency has steadily increased from 4 issues in 1988, to 6 issues in 1997 and to 12 issues in 2000. The total number of pages per volume and the number of original or review papers published also increased. Some significant milestones in the history of the AJAS include that (1) it became a Science Citation Index (SCI) journal in 1997, (2) the impact factor of the journal improved from 0.257 in 1999 to 0.446 in 2000, (3) it became a monthly journal (12 issues per volume) in 2000, (4) it adopted an English editing system in 1999, and (5) it has been covered in "Current Contents/Agriculture, Biology and Environmental Science since 2000. The AJAS is subscribed by 842 individuals or institutions. Annual subscription fees of US$ 50 (Category B) or US$ 70 (Category A) for individuals and US$ 70 (Category B) or US$ 120 (Category A) for institutions are much less than the actual production costs of US$ 130. A list of the 1,761 papers published in AJAS, listed according to subject area, may be found in the AJAS homepage (http://www.ajas.snu.ac.kr) and a very well prepared "Editorial Policy with Guide for Authors" is available in the Appendix of this paper. With regard to the submission status of manuscripts from AAAP member countries, India (235), Korea (235) and Japan (198) have submitted the most manuscripts. On the other hand, Mongolia, Nepal, and Papua New Guinea have never submitted any articles. The average time required from submission of a manuscript to printing in the AJAS has been reduced from 11 months in 1997-2000 to 7.8 months in 2001. The average rejection rate of manuscripts was 35.3%, a percentage slightly higher than most leading animal science journals. The total number of scientific papers published in the AJAS by AAAP member countries during a 14-year period (1988-2001) was 1,333 papers (75.7%) and that by non- AAAP member countries was 428 papers (24.3%). Japanese animal scientists have published the largest number of papers (397), followed by Korea (275), India (160), Bangladesh (111), Pakistan (85), Australia (71), Malaysia (59), China (53), Thailand (53), and Indonesia (34). It is regrettable that the Philippines (15), Vietnam (10), New Zealand (8), Nepal (2), Mongolia (0) and Papua New Guinea (0) have not actively participated in publishing papers in the AJAS. It is also interesting to note that the top 5 countries (Bangladesh, India, Japan, Korea and Pakistan) have published 1,028 papers in total indicating 77% of the total papers being published by AAAP animal scientists from Vol. 1 to 14 of the AJAS. The largest number of papers were published in the ruminant nutrition section (591 papers-44.3%), followed by the non-ruminant nutrition section (251 papers-18.8%), the animal reproduction section (153 papers-11.5%) and the animal breeding section (115 papers-8.6%). The largest portion of AJAS manuscripts was reviewed by Korean editors (44.3%), followed by Japanese editors (18.1%), Australian editors (6.0%) and Chinese editors (5.6%). Editors from the rest of the AAAP member countries have reviewed slightly less than 5% of the total AJAS manuscripts. It was regrettably noticed that editorial members representing Nepal (66.7%), Mongolia (50.0%), India (35.7%), Pakistan (25.0%), Papua New Guinea (25.0%), Malaysia (22.8%) and New Zealand (21.5%) have failed to return many of the manuscripts requested to be reviewed by the Editor-in-Chief. Financial records show that Korea has contributed the largest portion of production costs (68.5%), followed by Japan (17.3%), China (8.3%), and Australia (3.5%). It was found that 6 AAAP member countries have contributed less than 1% of the total production costs (Bangladesh, India, Indonesia, Malaysia, Papua New Guinea and Thailand), and another 6 AAAP member countries (Mongolia, Nepal and Pakistan, Philippine and Vietnam) have never provided any financial contribution in the form of subscriptions, page charges or reprints. It should be pointed out that most AAAP member countries have published more papers than their financial input with the exception of Korea and China. For example, Japan has published 29.8% of the total papers published in AJAS by AAAP member countries. However, Japan has contributed only 17.3% of total income. Similar trends could also be found in the case of Australia, Bangladesh, India, Indonesia, Malaysia and Thailand. A total of 12 Asian young animal scientists (under 40 years of age) have been awarded the AJAS-Purina Outstanding Research Award which was initiated in 1990 with a donation of US$ 2,000-3,000 by Mr. K. Y. Kim, President of Agribrands Purina Korea Inc. In order to improve the impact factor (citation frequency) and the financial structure of the AJAS, (1) submission of more manuscripts of good quality should be encouraged, (2) subscription rate of all AAAP member countries, especially Category B member countries should be dramatically increased, (3) a page charge policy and reprint ordering system should be applied to all AAAP member countries, and (4) all AAAP countries, especially Category A member countries should share more of the financial burden (advertisement revenue or support from public or private sector).

A Study on the Reducing Pollutants in Non-Ruminant Manure by Increasing Feed Utilization (사료이용율 증가에 따른 비반추가축의 분뇨에 의한 공해발생 감소에 관한 연구)

  • Nahm, K.H.
    • Korean Journal of Poultry Science
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    • v.28 no.3
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    • pp.245-257
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    • 2001
  • Localization of livestock facilities leads to concentration of livestock wastes and subsequent leakage of pollutants into the environment, resulting in public concern about their effects. Nitrogen (N) and phosphorus (P) are the most harmful components of animal manure, but odor from the manure itself and the livestock facilities is also a problem. Improving the nutrient efficiency of the livestock helps to decrease excretion of these environmental contaminants. Pigs and chickens are the main experimental models used in studies to improve nutrient efficiency. Addition of feed supplements and modifying feeding systems to improve nutrient efficiency can result in significant decrease in the N, P, odor and dry matter (DM) weight of manure. Examples of these methods include the following. 1) Addition of synthetic amino acids and reducing protein contents resulted N reductions of 10∼27% in broilers, 18∼35% in chicks and layers, 19∼62% in pigs, and a 9∼43% reduction in odor in pigs. 2) Enzyme supplementation resulted in a 12∼15% reduction in DM weight in broiler manure. 3) Phvtase supplementation resulted in P reductions of 25∼35% in chickens and 20∼60% in pigs. 4) Use of growth promoting substances resulted in a 5∼30% reduction in N and a 53∼56% reduction in odor of pigs. 5) Formulating diets closer to requirements (diet modification) reduced N and P by 10∼15% each in chickens and pigs, and odor by 28∼ 79% in pigs. 6) Phase feeding reduced N and P excretion by chicken and pigs from 10∼33% and 10∼13% each, as well as odor in growing and finishing pigs by 49∼79%. 7) Use of highly digestible raw materials in feed reduced N and P excretion by 5% in chickens and pigs.

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Milk Conjugated Linoleic Acid (CLA) Profile and Metabolic Responses of Dairy Cows Fed with High-temperature-micro-time (HTMT) Treated Diets Containing High Quantity Extruded Soybean (ESB)

  • Lee, H.G.;Hong, Z.S.;Wang, J.H.;Xu, C.X.;Jin, Y.C.;Kim, T.K.;Kim, Y.J.;Song, M.K.;Choi, Yun.-Jaei
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.11
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    • pp.1504-1512
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    • 2009
  • A feeding trial was conducted to examine the effect of high-temperature-micro-time (HTMT) processing of diets containing extruded soybean (ESB) in high quantity on milk fat production, metabolic responses, and the formation of conjugated linoleic acid (CLA) and trans-vaccenic acid (TVA). Twenty-one multiparous Holstein cows in mid-lactation were blocked according to milk yield in the previous lactation. Cows within each block were randomly assigned to either normal concentrate or HTMT treated diets containing ESB (7.5% HTMT-ESB and 15% HTMT-ESB). It was hypothesized that the HTMT-ESB would affect the undegradable fatty acids in the rumen and, thus, would modify the fatty acid profile of milk fat. Both 7.5% and 15% HTMT-ESB did not affect milk yield, fat, protein, lactose and solid-not-fat (SNF), but the proportion of cis-9, trans-11 CLA in milk fat was significantly increased by these treatments. Content of TVA in milk fat was not affected by HTMT-ESB. The HTMT-ESB influenced the fatty acid profile in milk fat, but there was little difference between 7.5% and 15% of supplementation. HTMT-ESB feeding significantly decreased the concentration of plasma insulin and glucose, while plasma growth hormone (GH), triglyceride (TG), non-esterified fatty acid (NEFA) and HDLcholesterol were increased by 7.5% and 15% ESB-HTMT supplementation in comparison to the control group (p<0.05). However, no significant difference was observed in plasma LDL-cholesterol, insulin like growth factor (IGF)-1, T3, T4, and leptin concentrations among treatments (p>0.05). The present results showed that cis-9, trans-11 CLA production was increased by HTMT treatment of dietary ESB without reduction of milk fat, and the unchanged milk fat and yield was assumed to be associated with the constant level of thyroid hormones, leptin, and IGF-1.

Fruit and vegetable discards preserved with sodium metabisulfite as a high-moisture ingredient in total mixed ration for ruminants: effect on in vitro ruminal fermentation and in vivo metabolism

  • Ahmadi, Farhad;Lee, Won Hee;Oh, Young-Kyoon;Park, Keunkyu;Kwak, Wan Sup
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.446-455
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    • 2020
  • Objective: Our recent series of laboratory- and large-scale experiments confirmed that under aerobic and anaerobic conditions, sodium metabisulfite (SMB) was effective in preserving nutrients and antioxidant capacity of highly perishable fruit and vegetable discards (FVD). Hence, the purpose of this study was to examine how partial inclusion of SMB-treated FVD in total mixed ration (TMR) influences in vitro ruminal fermentation, whole-tract digestibility, nitrogen metabolism, blood metabolites, and voluntary feed intake of sheep. Methods: The FVD were mixed thoroughly with 6 g SMB/kg wet biomass and kept outdoors under aerobic conditions for 7 days. Four TMRs including four levels of SMB-treated FVD (as-fed basis) at 0%, 10%, 20%, and 30% (equaling to 0%, 1.9%, 3.8%, and 5.7% on dry matter basis, respectively), were prepared as replacement for corn grain. The ruminal fermentation metabolites were studied using an in vitro gas production test. Four mature male Corriedale sheep were assigned at random to the 4 diets for two separate sub-experiments; i) digestibility trial with four 21-d periods, and ii) voluntary feed intake trial with four 28-d periods. Results: Inclusion of SMB-treated FVD in the TMR tended to quadratically increase partitioning factor. No effect was seen on total-tract digestibility of organic matter, ether extract, crude protein, and acid detergent fiber, except for neutral detergent fiber digestibility that tended to linearly increase with increasing SMB-treated FVD in the TMR. The progressive increase of FVD preserved with SMB in the diet had no effect on nitrogen metabolism. Treatment had no effect on serum antioxidant capacity and blood metabolites assayed. Voluntary feed intake was not impaired by inclusion of SMB-treated FVD in the TMR. Conclusion: It appears that FVD preserved with SMB can be safely incorporated into TMR as replacement of corn grain without impairment of nutrient metabolism and feed intake.

Nutrient Balance and Glucose Metabolism of Female Growing, Late Pregnant and Lactating Etawah Crossbred Goats

  • Astuti, D.A.;Sastradipradja, D.;Sutardi, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.8
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    • pp.1068-1075
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    • 2000
  • A study involving nutrient balances and radioisotope labeling techniques was undertaken to study energy and protein metabolism, and glucose kinetics of female crossbred Etawah goats, using 12 weaned (BW $14.0{\pm}2.0kg$), 12 late pregnant (BW $27.8{\pm}1.8kg$) and 12 first lactation does (BW $25.0{\pm}5.0kg$). Each class of animal was randomly allotted into 3 dietary treatment groups R1, R2 and R3, that received 100%, 85%, and 70% of ad libitum feed. The rations offered were pellets containing 21.8% CP and 19.3 MJ GE/kg, except for the lactating does who received pellets (17.2% CP and 18.9 MJ GE/kg) and fresh Penisetum purpureum grass. Energy and nitrogen balance studies were conducted during a two-week trial. Daily heat production (HP, estimated by the carbon dioxide entry rate technique), glucose pool and flux were measured. Equations were found for metabolizable energy (ME) and protein intake (IP) requirements for growing goats: ME (MJ/d)=1.87+0.55 RE-0.001 ADG+0.044 RP $(R^2=0.89)$ and IP (g/d)=48.47+2.99 RE+0.029 ADG+0.79 RP $(R^2=0.90)$; for pregnant does: ME (MJ/d)=5.92+0.96 RE-0.002 ADG+0.003 RP $(R^2=0.99)$ and IP (g/d)=58.34+5.41 RE+0.625 ADG-0.30 RP $(R^2=0.98)$; and for lactating does: ME (MJ/d)=4.23+0.713 RE+0.003 ADG+0.006 RP+0.002 MY $(R^2=0.86)$; IP (g/d)=84.05-5.36 RE+0.055 ADG-0.16 RP+0.068 MY $(R^2=0.45)$, where RE is retained energy (MJ/d), ADG is average daily gain in weight (g/d), RP is retained protein (g/d) and MY is milk yield (ml/d). ME and IP requirements for maintenance for growing goats were 0.46 MJ/d.kg $BW^{0.75}$ and 7.43 g/d.kg $BW^{0.75}$, respectively. Values for the pregnant and lactating does were in the same order, 0.55 MJ/d.kg $BW^{0.75}$ and 11.7 g/d.kg $BW^{0.75}$, and 0.50 MJ/d.kg $BW^{0.75}$ and 10.8 g/d.kg $BW^{0.75}$, respectively. Milk protein ranged from 3.06 to 3.5% and milk fat averaged 5.2%. Glucose metabolism in Etawah crossbred female goat is active, but glucose flux is low compared to temperate ruminant breeds which may implicate its role to support production.

Effect of optimal sodium stearoyl-2-lactylate supplementation on growth performance and blood and carcass characteristics in Hanwoo steers during the early fattening period

  • Lee, Shin Ja;Lee, Sang Suk;Kim, Eun Tae;Jeong, Jin Suk;Lee, Ji Hoon;Jeong, Joon;Park, Joong Kook;Park, Beom Young;Jeong, Ha Yeon;Ki, Kwang Seok;Kim, Chang Hyun;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1442-1448
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    • 2018
  • Objective: This study was conducted to evaluate the effect of different levels of total digestible nutrients (TDN) and sodium stearoyl-2-lactylate (SSL) supplementation on growth performance and blood and carcass characteristics in Hanwoo steers during the early fattening period. Methods: Sixty Hanwoo steers (average body weight, $333{\pm}36.4kg$) were randomly allotted to 3 treatments, with twenty steers per treatment, and ten steers per pen with a size of $80m^2$. Dietary treatments were as follows: CON, basal diet; treatment (TRT) 0.5, 0.5% down-spec of TDN with 0.1% SSL; TRT 1.0, 1.0% down-spec of TDN with 0.1% SSL. Results: The results demonstrated that average daily gain and feed efficiency increased with TRT 0.5 (0.85 kg and 11.68) vs CON (0.82 kg and 11.27) or TRT 1.0 (0.78 kg and 10.74), indicating that 0.1% SSL supplementation in the feed of early fattening steers may result in a saving of 0.5% TDN. No significant differences were observed amongst all treatments (p>0.05) for blood metabolite concentration and blood corpuscle values, which were all within the normally accepted range for healthy steers. Conclusion: Our study suggests that a TDN 0.5% down spec with 0.1% SSL supplemented feed may be effective and profitable for the early fattening period of Hanwoo steers without causing adverse effects.

Nutritive Evaluation of Poultry Waste and Sudex Grass Silage for Sheep

  • Chaudhry, S.M.;Naseer, Z.;Alkraidees, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.1
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    • pp.79-85
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    • 1997
  • Cage layer waste and sudex grass were ensiled in the proportions of 0:100, 30:70, 40:60 and 50:50 wet basis, respectively. The influence of ensiling cage layer waste on Salmonellae, Shigella, Proteus, and total number of colony forming units (CFU) was investigated. The nutritive value of the silages was evaluated in a digestion trial. The experiment was conducted with 24 wethers allotted to four silages. Initial samples of cage layer waste showed $0.11{\times}10^6$ CFU and salmonellae, Shigella and Proteus were present. Ensiling was effective in complete elimination of all the pathogens. Dry matter, crude protein and ash contents were increased linearly (p < .01) with the increase of cage layer waste in the silages ($358g{\cdot}kg^{-1}$ to $484g{\cdot}kg^{-1}$; $76.3g{\cdot}kg^{-1}$ to $183.2g{\cdot}kg^{-1}$ and $38.5g{\cdot}kg^{-1}$ to $169.4g{\cdot}kg^{-1}$; DM basis, respectively). Water solubles carbohydrate values for silages were 38.3, 22.5, 20.1 and $20.0g{\cdot}kg^{-1}$ DM basis, respectively. Ensiling decreased the pH values for all the silages and the decrease was higher for sudex grass ensiled alone than grass ensiled with cage layer waste. Lactic acid concentration in silages increased linearly (p < .01) with the increase of cage layer waste. The apparent digestibilities of DM, OM and CP for the animals fed sudex grass silage alone were 496.0, 516.1, $496.7g{\cdot}kg^{-1}$ DM basis respectively. However, digestibilities of all the components were higher (p < 0.01) for the animals fed silages containing cage layer waste. Among waste containing diets, the digestibilities of all the components showed quadratic affect (p < 0.01), increased for silages containing 30 and 40% cage layer waste and decreased when the level of waste increased from 40 to 50%. The results indicated that cage layer waste can be used upto 40% in ruminant diet as a source of N without any adverse effect on the health of animals. Ensiling appeared to be feasible and effective method for eliminating the pathogen present in cage layer waste.