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Tài liệu Sử dụng thân cây chuối làm thức ăn cho lợn địa phương (kandol) ở vùng núi ratanakiri của campuchia

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HUE UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY HUY SOKCHEA UTILIZATION OF BANANA STEMS FOR LOCAL PIGS (KANDOL) IN MOUNTAINOUS RATANAKIRI PROVINCE OF CAMBODIA DOCTOR OF PHILOSOPHY IN ANIMAL SCIENCES HUE, 2019 i HUE UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY HUY SOKCHEA UTILIZATION OF BANANA STEMS FOR LOCAL PIGS (KANDOL) IN MOUNTAINOUS RATANAKIRI PROVINCE OF CAMBODIA SPECIALIZATION: ANIMAL SCIENCES CODE: 9620105 DOCTOR OF PHILOSOPHY IN ANIMAL SCIENCES SUPERVISOR 1: ASSOC. PROF. TRAN THI THU HONG SUPERVISOR 2: PROF. LE DUC NGOAN HUE, 2019 ii GUARANTEE I hereby guarantee that scientific work in this thesis is mine. All results described in this thesis are righteous and objective. Two papers were published in Journal of Veterinary and Animal Research, one paper was in International Journal of Innovation and Animal Research. Hue, March 2019 Huy Sokchea, PhD student iii Acknowledgements I am very pleased to express my sincere and gratitude to institutions and individuals, who involved in and contributed to my doctoral thesis. Special thanks to the Swedish International Development Authority/Department for Research Cooperation (Sida/SAREC) for financially support of my researches and study in both Cambodia and Vietnam through the MEKARN program (Mekong Basin Animal Research Network) and also to my supervisors, Asso. Prof. Tran Thi Thu Hong for her constructive advices and useful guidance and also to my co-supervisor Prof. Le Duc Ngoan, Asso. Prof. Le Dinh Phung and H.E. Khieu Borin for their inputs in both experiments and the thesis. In addition, I would like to thank very much to Prof. Le Duc Ngoan, Asso. Prof. Le Van An, Asso. Prof. Le Dinh Phung, Asso. Prof. Nguyen Quang Linh and Asso. Prof. Nguyen Xuan Ba for providing the training courses on advanced method of writing academic papers; advanced livestock feed and feeding; advanced biology statistics and experiment design; advanced pig husbandry; and advanced cattle husbandry, respectively and also thank to the students from Royal University of Agriculture (Mr. Sao Kongkea and Thim Chan Thy) for helping me in running my experiments and staffs of CelAgrid (Dr. Chhay Ty, Dr. Miech Phalla, Dr. Chiv Phiny, Dr. Pok Samkol, Mrs. Bou Socheata, Mr. Son Pov, Mr. Vo Sina and Ms. Chourn Kimyeang) for their contributions during my thesis development. Finally, I also would like to convey my sincere gratitude to my wife, children, parents, parents in law, brothers, sisters and a brother and a sister in-law for their valuable encouragement and understanding. iv Abstract The overall objective of the study was to effectively utilize banana stems for improving local pigs performance under village conditions in the mountainous zones of Cambodia. In this thesis, four studies were performed to meet main specific objective. In the first study, nine villages of 3 communes and 3 districts with totally 126 respondents were sampled for this study in order to understand the situation of pig production of farmers in mountainous Ratanakiri province. As result, all famers preferred keeping local pigs in the range of 3-5 heads per family and the pigs were fed for 8-12 months to get the marketable weight of 30-40 kg (ADG 120g/day) with the diet composed of banana stems 3.8% as DM basic that consisted of 2,257 kcal ME/kg DM and 7.8% CP. In the second study, the experiment was followed by nested model with 3 replicates to determine the effects of time, C/N ratio and molasses concentration on yeast of S. cerevisiae biomass production. It was found that the application of C/N ratio at 10/1 as substrate for 24 hours was able to improve biomass production of Saccharomyces cerevisiae. In the third study, the experiment was designed, following to completely randomized design (CRD) with four treatments and 4 replicates in the purpose of improvement of nutritive values of banana stems by fermentation with Saccharomyces cerevisiae solution. As result, the fermentation of banana stems with the addition of Saccharomyces cerevisiae solution could improve their nutritive values, mainly true protein and crude fiber in the period of 7 days, compared to the ones without any addition of Saccharomyces cerevisiae solution. In the last study, the experiment was designed by the randomized completely design (RCD) with 5 dietary treatments and 4 replicates to determine the optimum inclusive level of fermented banana stems in the diets on apparent digestibility, growth performance and carcasses quality of local pigs. As result, the inclusion of fermented banana stems at the 50% into the diet could improve apparent digestibility and growth performance as mainly compared to the control diet, however, any inclusion of fermented banana stems into the diet was not quite effect on carcasses quality. i Key words: Carcasses quality, digestibility, growth performance, local pig, Saccharomyces cerevisiae fermented banana stems ii Dedication to My parents, parents in law, brothers and sisters My wife Pech Sina My children, Huy Soknancy, Huy Sokjulie and Huy Sokyannyheng iii TABLE OF CONTENT INTRODUCTION.....................................................................................................................10 1. Problem statement.........................................................................................................10 2. Overall and specific objectives of the study.................................................................11 2.1.1. Overall objective............................................................................................................11 2.1.2. Specific objectives..........................................................................................................11 3. Significant/Innovation of the dissertation....................................................................12 CHAPTER 1..............................................................................................................................13 LITERATURE REVIEW.........................................................................................................13 1. Pig production in Cambodia.........................................................................................13 1.1. Pig production systems..................................................................................................13 1.2. Pig production in mountainous zones..........................................................................14 2. Fibrous feeds for pigs....................................................................................................15 2.1. Roles of fibrous feeds.....................................................................................................15 2.2. Fractions of fibrous feeds..............................................................................................16 2.2.1. Cellulose.........................................................................................................................16 2.2.2. Hemicellulose.................................................................................................................16 2.2.3. Lignin.............................................................................................................................17 2.3. Fibrous feed movement process in the digestive tract of the pigs..............................17 2.4. Effect of fibrous feeds on intake...................................................................................17 2.5. Effect of fibrous feeds in pig diet..................................................................................18 2.5.1. Digestibility....................................................................................................................18 2.5.2. The digestive tracts’ health...........................................................................................19 2.5.3. Growth performance.....................................................................................................20 2.5.4. Carcass quality..............................................................................................................21 3. Available and local fibrous feed resources...................................................................21 3.1. Banana pseudo stems.....................................................................................................22 3.1.1. Nutritive value...............................................................................................................22 3.1.2. Utilization.......................................................................................................................22 3.2. Rice bran........................................................................................................................23 1 3.2.1. Nutritive value...............................................................................................................23 3.2.2. Utilization.......................................................................................................................24 3.3. Rice distiller’s by-product.............................................................................................25 3.3.1. Nutritive value...............................................................................................................25 3.3.2. Utilization.......................................................................................................................25 4. Yeast fermentation................................................................................................................26 4.1. Fermentation..................................................................................................................26 4.2. Factors affect Saccharomyces cerevisiae biomass........................................................27 4.2.1. Carbon and nitrogen ratios...........................................................................................27 4.2.2. Temperature and pH.....................................................................................................28 4.3. Effect of microorganism and organic acids on animal production and health.........28 4.3.1. Saccharomyces cerevisiae yeast.....................................................................................28 4.3.2. Enzyme and organic acids.............................................................................................29 4.4. Solid state fermentation................................................................................................29 4.4.1. Concept...........................................................................................................................29 4.4.2. Solid state fermention of agricultural byproducts......................................................30 4.4.3. Advantages and disadvantages.....................................................................................30 References..................................................................................................................................31 CHAPTER 2..............................................................................................................................48 UNDERSTANDING PIG PRODUCTION IN RATANAKIRI PROVINCE........................48 Abstract......................................................................................................................................48 1. Introduction.........................................................................................................................49 2. Methodologies........................................................................................................................50 2.1. Site selection and duration............................................................................................50 2.2. Sample size.....................................................................................................................50 3. Statistical analysis..................................................................................................................51 4. Results and discussion...........................................................................................................51 4.1. General information about targeted sites....................................................................51 4.2. Family profile.................................................................................................................52 4.3. Banana information.......................................................................................................53 4.4. Livestock information...................................................................................................53 4.5. Pig raising system..........................................................................................................54 2 4.6. Pig production and health.............................................................................................55 4.7. Pigs breed and breeding................................................................................................56 4.8. Feed and feeding systems..............................................................................................57 4.9. Diseases and prevention................................................................................................60 4.10. Pigs market demand......................................................................................................60 4.11. Problems and solutions on pig production...................................................................60 5. Conclusion..............................................................................................................................61 References..................................................................................................................................61 CHAPTER 3..............................................................................................................................65 EFFECT OF TIME, C/N RATIO AND MOLASSES CONCENTRATION ON SACCHAROMYCES CEREVISIAE BIOMASS PRODUCTION..........................................65 Abstract......................................................................................................................................65 1. Introduction.........................................................................................................................66 2. Material and methods.........................................................................................................67 2.1. Yeast strain and preparation........................................................................................67 2.2. Experimental design......................................................................................................67 2.3. Measurements................................................................................................................68 2.3.1. Yeast density..................................................................................................................68 2.3.2. Identification of yeast....................................................................................................69 2.3.3. PCR amplification of the ITS region............................................................................69 2.4. Cultivation process of yeast..........................................................................................70 2.5. Analytical procedures....................................................................................................70 2.6. Data analysis..................................................................................................................70 3. Results..................................................................................................................................71 3.1. Density of yeast..............................................................................................................71 3.2. Identification of yeast....................................................................................................71 3.3. Effect of time on Saccharomyces cerevisiae biomass...................................................72 3.4. Effect of C/N ratio on Saccharomyces cerevisiae biomass...........................................73 3.5. Effect of molasses concentration on Saccharomyces cerevisiae biomass....................74 4. Discussion............................................................................................................................75 5. Conclusion...........................................................................................................................78 References..................................................................................................................................78 3 CHAPTER 4..............................................................................................................................82 IMPROVING NUTRITIVE VALUES OF BANANA STEMS BY SACCHAROMYCES CEREVISIAE SOLUTION FERMENTATION......................................................................82 Abstract......................................................................................................................................82 1. Introduction.........................................................................................................................83 2. Materials and methods.......................................................................................................84 2.1. Experiment I..................................................................................................................84 2.1.1. Location..........................................................................................................................84 2.1.2. Experimental design......................................................................................................85 2.1.3. Banana stems fermenting process................................................................................85 2.1.4. Chemical analyses..........................................................................................................86 2.1.5. Statistical analyses.........................................................................................................86 2.2. Experiment II.................................................................................................................87 2.2.1. Experimental design......................................................................................................87 2.2.2. Statistical analysis..........................................................................................................87 3. Results and discussion........................................................................................................87 3.1. Experiment I..................................................................................................................87 3.1.1. Results............................................................................................................................87 3.1.2. Discussion.......................................................................................................................91 3.2. Experiment II.................................................................................................................93 3.2.1. Results............................................................................................................................93 3.2.2. Discussion.......................................................................................................................96 4. Conclusion......................................................................................................................97 References..................................................................................................................................97 CHAPTER 5............................................................................................................................102 EFFECT OF INCLUSION OF FERMENTED BANANA STEMS IN DIETS ON DIGESTIBILITY, GROWTH PERFORMANCE AND CARCASS QUALITY OF LOCAL PIG (KANDOL).......................................................................................................................102 Abstract....................................................................................................................................102 1. Introduction........................................................................................................................103 2. Materials and methods.....................................................................................................104 2.1. Location........................................................................................................................104 4 2.2. Preparation for Saccharomyces cerevisiae banana stem fermentation....................104 2.3. Experimental designs..................................................................................................105 2.4. Diets and feeding..........................................................................................................106 2.5. Sample collection of digestibility and feeding trial....................................................107 2.6. Chemical analyses........................................................................................................108 2.7. Statistical analyses.......................................................................................................109 3. Results and discussion.................................................................................................109 3.1. Apparent digestibility and nitrogen balance.............................................................109 3.1.1. Results..........................................................................................................................109 3.1.2. Discussion.....................................................................................................................110 3.2. Growth performance...................................................................................................111 3.2.1. Results..........................................................................................................................111 3.2.2. Discussion.....................................................................................................................114 4. Conclusion....................................................................................................................116 References................................................................................................................................116 CHAPTER 6............................................................................................................................122 GENERAL DISCUSSION AND CONCLUSIONS...............................................................122 1. GENERAL DISCUSSION..........................................................................................122 1.1. Situation of local pigs’ production in Cambodia.......................................................122 1.2. Saccharomyces cerevisiae and its application.............................................................123 1.3. Nutritive value improvement of banana stems through fermentation.....................124 1.4. Banana stems-local pigs’ production system.............................................................125 1.5. Improving of banana stems by fermentation for local pigs’ production.................126 2. GENERAL CONCLUSIONS.....................................................................................131 3. IMPLICATIONS AND FURTHER RESEARCH....................................................131 3.1. Implications..................................................................................................................131 3.2. Further research..........................................................................................................132 REFERENCES............................................................................................................................132 5 LIST OF TABLES Table 1: Proximate composition of rice bran (% air dry weight basis)...........................................24 Table 2: Sampling of each village in Ratanakiri.............................................................................51 Table 3: Family profile by commune/district.................................................................................52 Table 4: Animal population by communes.....................................................................................54 Table 5: Average number of animals per household......................................................................54 Table 6: Feed resources for local pig in Ratanakiri (% of interviewed householder).....................57 Table 7: Chemical composition of the feeds in DM basic..............................................................58 Table 8: Estimated amount of feed intake (g DM/day), ME intake and nutritive value of the diet. 59 Table 9: The medium for yeast fermentation..................................................................................68 Table 10: Density of yeast in feed active dry yeast product...........................................................71 Table 11: Gene sequences of DNA bands from yeast detected by Gene bank................................72 Table 12: Effect of time on Saccharomyces cerevisiae biomass (g L-1)..........................................73 Table 13: Effect of C/N ratio on Saccharomyces cerevisiae biomass (g L-1)..................................73 Table 14: Formulation of fermentation of banana stem and rice bran in DM basis........................85 Table 15: Chemical composition of the feeds in DM basic............................................................85 Table 16: Chemical composition of fermented banana stem at the different treatments.................88 Table 17: Chemical composition of banana stem and rice bran (DM basic)..................................93 Table 18: Chemical composition of fermented banana stem at the different treatments.................93 Table 19: Chemical composition of ingredients used in the diets.................................................106 Table 20: Diets for digestibility and feeding study (% DM).........................................................106 Table 21: Apparent digestibility of treatmentary diets (%)...........................................................109 Table 22: Nitrogen balance of the pigs in different treatments.....................................................110 Table 23: Mean value of feed intake of pig in different treatments..............................................111 Table 24: Average daily gain and feed conversion ratio of pigs in different treatments...............111 Table 25: Mean value of carcass traits of pig fed different treatmentary diets..............................112 Table 26: Back fat thickness (mm) and loin eye area (cm 2) of pigs in different treatments..........112 Table 27: Mean value of pH, color, marbling and water holding capacity of meat in treatments. 113 6 LIST OF FIGURES Figure1: Non-starch polysaccharide components (Choct et al., 2010)...........................................16 Figure 2: Mean of S. cerevisiae biomass at C/N ratio of 5/1..........................................................75 Figure 3: Mean of S. cerevisiae biomass at C/N ratio of 10/1........................................................75 Figure 4: Mean of S. cerevisiae biomass at C/N ratio of 15/1........................................................75 Figure 5: Effect of fermented times at each treatment on DM........................................................90 Figure 6: Effect of fermented times at each treatment on ash.........................................................90 Figure 7: Effect of fermented times at each treatment on CF.........................................................90 Figure 8: Effect of fermented times at each treatment on CP.........................................................91 Figure 9: Effect of fermented times at each treatment on TP.........................................................91 Figure 10: Effect of fermented times at each treatment on DM......................................................95 Figure 11: Effect of fermented times at each treatment on ash.......................................................95 Figure 12: Effect of fermented times at each treatment on CP.......................................................95 Figure 13: Effect of fermented times at each treatment on TP.......................................................96 7 LIST OF PHOTOS Photo 1: Banana plants farming in upland......................................................................................53 Photo 2: Jungle banana plants........................................................................................................53 Photo 3: Banana stems cutting........................................................................................................59 Photo 4: Broken rice cooking.........................................................................................................59 Photo 5: DNA band of three DNA samples from 3 colonies (A-C) of yeast on agarose gels.........72 Photo 6: Banana farming..............................................................................................................105 Photo 7: Collected banana stems..................................................................................................105 Photo 8 &9: View of the pens for digestibility study...................................................................107 Photo 10: Measurement of back fat thickness..............................................................................113 Photo 11: Measurement of loin eye area......................................................................................113 Photo 12: Color and marbling score.............................................................................................114 Photo 13: Measurement of meat pH.............................................................................................114 8 LIST OF ABBRIVIATIONS ADB Asian Development Bank CF Crude Fiber CP Crude Protein DM Dry Matter FAO Food and Agriculture Organization FCR Feed Conversion Ratio GDAHP General Directorate of Animal Health and Production GDP Gross Domestic Product MAFF Ministry of Agriculture and Forestry and Fishery NIS National Institute of Statistic OM Organic Matter PDAFF Provincial department of agriculture, forestry and fishery TP True Protein VAHWs Village Animal Health Workers 9 INTRODUCTION 1. Problem statement Cambodia population is over 16 million, annually growth rate of 1.5% and population density of about 91 habita per km 2 in 2018. Agriculture is one of three economic sectors which contributed 26.3 % of national GDP in 2016. Livestock sub-sector contributed about 2.8% to the national economy and 11.96% of agricultural GDP (MAFF, 2017). However, pig production shared about 48% of the total livestock production (FAO, 2011). In 2016, pig population was reached 2,970,624 heads (annually growth rate 2%), and 98% of pig population produced by smallholders (MAFF, 2018). About 5% of smallholders raised local breeds (Borin et al., 2012), due to more resistant to infectious diseases and more adapting to local climate frequency and environment and they also have a higher capacity to digest higher fibrous feed and more reproductive than exotic ones (Rodríguez and Preston, 1997; Len et al., 2009b). The smallholders have turned in a very low level of production due to low input and lack of technical knowledges and experiences in animal husbandry (Wallberg et al., 2011). Total of 54.7% of smallholders fed their pigs with local resources and agricultural byproducts (Borin et al., 2012). Ström et al. (2017) indicated that partly and/or fully utilization of the unconventional feeds was potential and sustainable alternative of pigs’ production, mainly for smallholder pigs’ production as it could alleviate the production cost and risks. Banana was one of the fruit trees with totally cultivated land of 24,000ha after mango of 42,000ha, produced over 240,000 tons of fruits and 960,000 tons of residues yearly in Cambodia (NIS, 2015). Seven of the 25 provinces in Cambodia reported to plant banana on at least 1,000 ha. Kampong Cham led other provinces with 5,000 ha of banana plantation and with 819 ha in Ratanakiri (data from provincial department of agriculture in 2019), but only 350 ha in 2013 (NIS, 2015). With these cultivated areas, about 3,300 tons of banana stems residue remained in the field with improperly and sufficiently utilization. In addition, Jungle banana plants grew naturally for year-rounds in somewhere around the 10 farms or sometimes in the farms and along the ways to the farms as well, so it was very hard to estimated how many hectares of the banana plants were. However, they were only harvested as feed of the pigs in some months of the year, especially in the period of cultivated banana plants and other feed resources were in minority or extinction. Wang et al. (2016) reported that banana stems could be considered as an alternative of traditional forage sources. Pigs are generally fed in fresh basic by chopped and pasted together with rice bran, plus some cooked rice or/and kitchen waste (Wallberg et al., 2011). Chhay ty et al. (2016) showed that banana stems has very low dry matter and nutritive values, but it could be improved by fermentation with indigenous microorganism (IMO) (Michael et al., 2016). Duyet et al. (2013) also found that 50:50 mixtures (DM basis) of banana stems and taro foliage could be ensiled satisfactorily without any additions. In addition, supplementation with a combination of 3% nitrogen, 0.4% sulphur and 0.25% phosphorus produced the highest protein content up to 8.98% which higher than the control one of 4.91% (Rochana et al., 2017). 2. Overall and specific objectives of the study 2.1.1. Overall objective To utilize banana stems effectively for improvement of local pig performance under mountainous village conditions in Cambodia. 2.1.2. Specific objectives The specific objectives of the study were:  To understand the situation of pig production of ethnic farmers in mountainous Ratanakiri province;  To determine the effects of time, C/N ratio and molasses concentration on yeast of Saccharomyces cerevisiae biomass production; 11  To improve nutritive values of banana stems through the fermentation with the Saccharomyces cerevisiae solution; and  To determine the optimum inclusive level of fermented banana stems in diets on apparent digestibility, growth performance and carcass quality of local pig. 3. Significant/Innovation of the dissertation The improvement of nutritive values of banana stems through fermentation with Saccharomyces cerevisiae for the feed of local pigs is acceptable alternative to the farmers, mainly those who living in mountainous areas in Cambodia. All papers in this thesis were firstly published in Cambodia. 12 CHAPTER 1 LITERATURE REVIEW 1. Pig production in Cambodia Pig population were totally 2,970,624 heads and it accounted for 98 percent of all small livestock raised in 2016 (MAFF, 2018). The highest density of 78.9 head/km 2 was found in Prey Veng Province and the lowest density of 0.74 head/km 2 in Koh Kong, however the density in Ratanakiri province was also low of only 2.70 heads/km2 (Sitha, 2012). The pork consumption is gradually increasing about 9.29 kg per capita (FAO, 2012), following to the population. Cambodian people needed about 894,108 head pigs a year in 2018, but the local production did not meet the demand, so the Ministry of Agriculture, Forestry and Fishery (MAFF) officially permitted 1,250 pigs to be imported from Vietnam and Thailand. 1.1. Pig production systems Pig production is categorized into three systems such as household/backyard, semi- intensive and intensive (MAFF, 2018). This is quite same as in Thailand, Vietnam, Lao, Myanmar and Philippines (FAO, 2011). Household production accounted for 76% (MAFF, 2018) and most of them were interested in the fattening pigs, a few kept the pigs for breeding purposes (NIS, 2015). They mostly utilized locally available resources such as rice bran, rice distiller by-product, vegetable wastes, cooked rice, kitchen waste and some concentrate feed for their pigs (Ström et al., 2017). By utilization of these local feed resources, the production cycle was longer from 7-10 months to get the weight from 70100 kg (Tornimbene et al., 2012), than semi-intensive system that was only 5.5 months averagely to get from 93-95kg by using homemade feed from cereal grain or using some concentrate. In addition, most of the household pig producers relied mainly on village animal health workers (VAHWs) for the vaccination and the treatment of their pigs and 13
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