Luận án Utilization of banana stems for local pigs (kandol) in mountainous ratanakiri province of cambodia

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
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
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
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. 
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. 
Key words: Carcasses quality, digestibility, growth performance, local pig, Saccharomyces cerevisiae fermented banana stems 
Dedication to
My parents, parents in law, brothers and sisters
My wife Pech Sina
My children, Huy Soknancy, Huy Sokjulie and Huy Sokyannyheng
TABLE OF CONTENT
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 (cm2) of pigs in different treatments	112
Table 27: Mean value of pH, color, marbling and water holding capacity of meat in treatments	113
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
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
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	
INTRODUCTION
Problem statement
Cambodia population is over 16 million, annually growth rate of 1.5% and population density of about 91 habita per km2 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 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). 
Overall and specific objectives of the study
Overall objective
To utilize banana stems effectively for improvement of local pig performance under mountainous village conditions in Cambodia.
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; 
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.
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. 
CHAPTER 1 
LITERATURE REVIEW
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/km2 was found in Prey Veng Province and the lowest density of 0.74 head/km2 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. 
 	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 70-100 kg (Tornimbene et al., 2012), than semi-intensive system that was only 5.5 months averagely to ... ggested to use for local pigs’ improvement as it was potential and economical feed and also available locally. However, it did not affect on hanging and dressing carcasses, and color, marbling and water holding capacity of the meat and viscera, small and large intestine.
IMPLICATIONS AND FURTHER RESEARCH 
Implications 
	The developed strategy in this thesis was for the improvement of nutritive value of available local feed resources and of the local pigs’ productivity by two concepts: (i) higher local pigs’ productivity and lower investment, mainly on feed; (ii) improvement of nutritive values of banana stems by fermentation with Saccharomyces cerevisiae.
 	Increasing local pigs’ productivity required feeding of highly conventional fibrous feed as sources of protein. Through the researches in this thesis has shown that the banana stems could be potentially and economically optional for the local pigs’ production of the ethnic people in the mountainous areas of Cambodia. The banana stems were fibrous feedstuff with lower nutritive value and higher water content. If feeding the pigs with the fresh ones would lead low intake due to many negative factors of higher fiber, antinutritional factors, palatability and water content, but these could be improved by fermentation with additive of rice bran and Saccharomyces cerevisiae. Finally, the thesis confirmed that valuable roles of fermented banana stems as the fibrous feedstuff were very important in improving local pigs’ production of the people in mountainous areas, mainly in community protected areas (CPA) of Cambodia.
Further research
	Further researches should be focused on some areas: (i) possible ways of promotion of local pigs’ production and of local feed resources, (ii) modifying the fermentation of banana stems by acceptable methodologies and additives and (iii) utilization of conventional banana stems for local pigs in rural areas, especially in community protected areas (CPA) of Cambodia. 
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