Livestock plays a important function in bettering living conditions of the hapless. The intent of farm animal maintaining of small-scale husbandmans in the Southern part is for the draught power, nutrient, hard currency income, and organic fertiliser from fecal matters ( Samkol and Lukefahr, 2008 ) . Recent eruptions of Avian Influenza in birds have occurred in several states in the universe, such as in Italy, Spain, and Hong Kong. One survey in Indonesia showed that 76 % of human extremely infective avian grippe ( HPAI ) H5N1 instances were proven to be associated with domestic fowl contact ( Sedyaningsih et al. , 2007 ) . Other surveies in Thailand, Vietnam, and China have suggested the association of human infection and domestic fowl eruptions with several hazard factors, including demographic features of worlds and animate beings, i.e. , human denseness and domestic fowl denseness ( Gilbert et al. , 2008 and Tiensin et al. , 2009 ) .
So, Rabbit production is a new development in the part, which plays an of import function in position of the economic hazards by the spread of Asiatic bird grippe ( Otte et al. , 2007 ) . Harmonizing to the FAO ( 2001 ) , backyard coney maintaining provides extra income and supplies extra protein for hapless rural and urban families with low investing and labour inputs. Rabbits have little organic structure size, short coevals interval, high generative potency, rapid growing rate, familial diverseness, and the ability to use eatages and byproducts as major diet constituents that make the carnal appropriate for little farm animal maintaining in developing states ( Cheeke, 1986 ) . Nowadays, coney genteelness has an increasing impact due to the nutritionary value of meat and economic traits of engendering. Development in this animate being industry besides induces to seek for new engineerings to better coney wellness and public assistance.
The spread of enteral diseases and in peculiar epizootic coney enteropathy has negatively affected the wellness position and caused lessening in carnal perfomance in coney farms and mostly increased antibiotics use as a preventative method ( Duperray et al. , 2003 ) . However, recent issues with bacterial opposition to these antibiotics urged scientists to happen options to the usage of antibiotics ( Smith et al. , 2002 ) . In add-on, on the base of consumer demand, European statute law is censoring the usage of antibiotics as growing boosters and is working towards a decrease of curative antibiotics for all farm animal production, to avoid crossed opposition in worlds and better nutrient safety.
Therefore, Probiotics appears as possible alternate provender additives to modulate enteric microflora and better animate being wellness ( Medina et al. , 2002 ) . However, surveies demoing the effects of combination of different probiotics trains on intestine wellness and growing of coneies are besides limited.
OBJECTIVES OF STUDY
The aims of this survey are probe the effects of Lacto-bacillus in digestibleness, ceacal agitation, microbic protein synthesis and growing public presentation of ablactating coneies.
KEYWORDS ( ENGLISH )
Weaning coneies, digestibleness, ceacal agitation, growing public presentation
The digestive physiology of the coney
The digestive system of the coney
The digestive system of the coney is characterized by the comparative importance of the cecum and colon when compared with other species ( Portsmouth, 1977 ) . As a effect, the microbic activity of the cecum is of great importance for the procedures of digestion and alimentary use, but besides in the control of digestive pathologies. Furthermore, caecotrophy, the behavior of consumption of soft fecal matters of cecal beginning, makes microbic digestion in the cecum more of import for the overall use of foods by the coney ( Carlos de Blas and Julian Wiseman, 2010 ) .
Stomach: the first of import compartment of the digestive system of the coney is the tummy ; this has a really weak muscular bed and is ever partly filled. The tummy pH ranges from 1 to 5, depending on site of finding, the presence or absence of soft fecal matters, the clip from feed consumption and the age of the coney ( Carlos de Blas and Julian Wiseman, 2010 ) .
Small bowel: the little bowel is the site where the greater portion of digestion and soaking up take topographic point by passive or active transit throughout the mucous membrane. Digestibility at the terminal of the ileum histories for 0.8-1 of the entire dietetic amino acid and amylum digestibleness ( Carabano et al. , 2009 ) . The pH of the little bowel is near to 7 ( Nicodemus et al. , 2002 ) .
Large bowel: The cecum is characterized by a weak muscular bed and contents. The cecal contents are somewhat acerb ( pH 5.4-6.8 ) ( Garcia et al. , 2002 ) . Gut-associated lymphoid tissue ( GALT ) and specialised cells regulate the interaction of the intestine mucous membrane with the microbiota and develop the mechanisms of tolerance and protection against pathogens.
Age-related alterations in the morphology and map of the digestive system
The tummy secretory organs are apparent in late foetuss ( 26 yearss ‘ gestation ) and true villi and enteric secretory organs are observed at 29 yearss ‘ gestation. During the suckling period, the mucosal secretory organs are able to bring forth enzymes to digest the chief constituents of the milk. Lactase activity is highest until 25 yearss of age and invertase and maltase rise until making the grownup degree at around 28-32 yearss ( Gallois et al. , 2008b ) . At circular 18 yearss of age the suckling coney begins to eat solid nutrient and diminish its milk consumption and the cecum and colon develop faster than the remainder of the digestive piece of land ( Gallois et al. , 2005 ) . From 3 to 7 hebdomads of age the cecum is filled by digesta and microbiota, and its contents reach a extremum at 7-9 hebdomads of age. The pH of the cecum is besides affected by age and lessenings from 6.8 at 15 yearss of age to 5.6 at 50 yearss of age ( Padilha et al. , 1995 ) .
The function of the enteric vegetation in the digestion and soaking up of foods
The presence of the microbic population in the cecum, together with caecotrophy, permits the coney to obtain extra energy, aminic acids and vitamins. The chief genus of the microbic population in the cecum of the grownup coney is Bacteroides ( Gouet and Fonty, 1973 ) , which comprises 109-1010 bacteriums g?1. During the first hebdomad of age, the digestive system of the coney is colonized by rigorous anaerobes, preponderantly Bacteroides. At 15 yearss of age, the Numberss of amylolytic bacteriums seem to stabilise, whereas those of colibacilli lessening as the Numberss of cellulolytic bacteriums addition ( Padilha et al. , 1995 ) . As a consequence of the fermentative activity of the microflora, VFAs are produced in the proportion of 60-80 mol of ethanoate, 8-20 mol of butyrate and 3-10 mol of propionate 100 mol?1 of VFAs ( Garcia et al. , 2002 ) . However, these proportions change with additions in the acetate concentration from 15 to 25 yearss of age and a reversal of the propionate to butyrate ratio from 15 to 29 yearss of age ( Padilha et al. , 1995 ) .
The consequence obtained Gidenne and Bellier ( 1992 ) in grownup animate beings showed that the VFA concentration in cecal contents depends on the clip of eating, lifting to a maximal 5 H after feeding. Cecal VFA concentrations are greater during the difficult fecal matters than during the soft fecal matters elimination period. Harmonizing to Bellier et Al. ( 1995 ) , this increase could hold two causes: ( I ) the greater flow of substrate to the cecum related to an addition in feed consumption during this period ; and ( two ) enrichment of the microbic population as a effect of antiperistaltic motions of the proximal colon. Cecal pH varies reciprocally to the addition in VFA concentration. Smaller values of cecal pH have been observed during difficult fecal matters elimination.
Rate of transition
The transition of provender through the tummy of the coney and cecum is comparatively slow and varies between 3-6 and 4-9 H, severally ( Gidenne and Poncet, 1985 ) . However, theodolite is really fast in the little bowel. Estimated keeping times in the jejunum and ileum are 10-20 and 30-60 proceedingss, severally ( Lebas, 1979 ) . Taking into history the full digestive piece of land, the average keeping clip varies from 9 to 30 H ( Laplace and Lebas, 1977 ) .
Alimentary demand of the growing coney
Table 1: Food demands of intensively coneies, as concentration kg?1 corrected to a dry affair content of 900 g kg?1
Unit of measurements
Lebas et al. , 1986
Lebas et al. , 1986
Methionine + Cysteine
Introduction about probiotics
Definition of probiotics
A probiotic is by and large dei¬?ned as a unrecorded microbial nutrient addendum which benei¬?cially affects the host by bettering its enteric microbic balance ( Fuller, 1989 ) . Microorganisms used in carnal provender are chiefly bacterial strains belonging to different genera, e.g. Bacillus ( B. Cereus, volt-ampere. toyoi, B. licheniformis, B. subtilis ) , Lactobacillus ( L. acidophilus, L. casei, L. farciminis, L. plantarum, L. rhamnosus ) , Enterococcus ( E. faecium ) and Pediococcus ( P. acidilactici ) . Other probiotics are microscopic Fungis, including Saccharomyces barms. Some probiotic micro-organisms are normal occupants in the digestive piece of land, while others are non ( Guillot, 2009 ) .
Some bacterial strains as probiotics
Lactic acid bacteriums ( add mentions )
Lactic acid bacteriums ( LAB ) are non-sporeaˆ?forming rods and coccus that agitation carbohydrates organizing lactic acid as the major endaˆ?product. Phylogenetically, the LAB belong to the clostridial subdivision of the gramaˆ?positive bacteriums, includes genera such as Clostridium, Bacillus, Listeria, and Staphylococcus ( Aguirre and Collins, 1993 ) . However, the term lactic acid bacterium has become normally associated with the genera Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, and Streptococcus. Decrease of pH and remotion of big sums of saccharides by agitation were considered the primary actions by which LAB inhibit foodaˆ?borne pathogens. LAB are capable of bring forthing repressive substances other, even at infrigidation temperatures with no growing than organic acids with repressive activity to diferent micro-organisms ( Amezquita and Brashears, 2000 ) .
To provide in animate being as probiotics, LAB must be able to last and metabolise in the bowel by resistibility to conditions encountered in the GI piece of land. Similarly, opposition of LAB to bile is an of import feature that enables them to last and turn in the enteric piece of land. Bile come ining the duodenal subdivision of the little bowel has been found to cut down endurance of bacteriums, likely because all bacteriums have cell membranes dwelling of lipoids and fatty acid, which are really susceptible to devastation by gall salts. Gilliland et Al. ( 1984 ) . In some surveies, it has been suggested that the ability of L. acidophilus to do a signii¬?cant addition in Numberss of Lactobacillus in the enteric piece of land may be critical for commanding growing of enteric pathogens. Hence, the success of a probiotics besides depend on the selected strain possessing bileaˆ?resistant qualities.
In the past few old ages, the LAB most normally associated with antibiotic opposition have been strains of the genera Enterococcus, particularly Enterococcus faecalis and E. faecium ( Franz et al. , 1999 ) .
The species that have been most extensively examined these are Bacillus subtilis, Bacillus clausii, Bacillus Cereus, Bacillus coagulans and Bacillus licheniformis. Bacillus are capable of bring forthing spore coats that afford protection from heat, enzymatic debasement, and the acidic conditions of the tummy ( Hong et al. , 2005 ) . Spores being heat-stable have a figure of advantages over other non-spore formers ( Simon M. Cutting, 2010 ) . The spore is capable of lasting the low pH of the stomachic barrier ( Barbosa et al. , 2005 ) so in rule a specii¬?ed dosage of spores can be stored indei¬?nitely without infrigidation and the full dosage of ingested bacteriums will make the little bowel integral.
Bacillus probiotic for carnal nutrition such as calves, domestic fowl, coneies and swine normally used Toyocerin® ( B. Cereus volt-ampere. toyoi ( NCIMB-40112/CNCM-1012 ) that is produced by Asahi Vet S.A. , Tokyo ( Head Off. ) , Japan ( hypertext transfer protocol: //www.asahi-kasei.co.jp ) . In the coneies, Hattori et Al. ( 1984 ) observed increased organic structure weight and a significant decrease of diarrhea when increasing the inclusion rate of B. Cereus volt-ampere. toyoi from 1 x 105 to 5 tens 106 spores/g diet. The supplementation of B. Cereus volt-ampere. toyoi at 2×105 spores/g diet in coney does and suckling kits improved growing public presentation and reduced the morbidity, gave higher values for litter weight and litter size at ablactating ( 25 vitamin D ) . ( Trocino et al. , 2005 ) . Collado et al. , 2007 indicated that combination of probiotics trains should see to increase the good wellness effects pompared with single strains, because of their interactive adhesion effecrs.
Lactobacillus ( Lactic acid bacteriums )
Lactobacilli are found in the normal enteric microbiota of animate beings and worlds ( Schneider et al. , 2004 ) , and have been identii¬?ed as responsible for commanding some effects of pathogens. Some lactic acid bacteriums ( LAB ) are capable of colonising the enteric piece of land of mice, staying in it without impacting provender consumption, and protecting animate beings against Salmonella Dublin DSPV 595T ( Frizzo et al. , 2006 ) . In add-on, LAB can colonise the GI piece of land of calves without translocating to the internal variety meats ( Frizzo et al. , 2009 ) . LAB could play a function in specific age groups, specifically neonates or the aged ( L.M.T. Dicks and M. Botes, 2010 ) .
Lactobacillus casei and Lactobacillus rhamnosus have been associated with bacteriemia and endocarditis ( Cannon et al. , 2005 ) . A few strains of LAB have been associated with urinary piece of land infections, lesion, tissue and other infections ( Vankerckhoven et al. , 2008 ) . In immunocom promised patients some lactobacilli have been associated with arthritis and meningitis ( Vankerckhoven et al. , 2008 ) .
The effects of probiotics in farm animal
Mode of action of probiotics
The benei¬?cial manners of action include: ( I ) decrease of toxin production ; ( two ) stimulation of enzyme production by the host ; ( three ) production of some vitamins or antimicrobic substances ; ( four ) competition for adhesion to epithelial cells and increased opposition to colonisation ; and ( V ) stimulation of the immune system of the host ( Fal & A ; ccedil ; ao-e-Cunha et al. , 2007 ) .
A figure of surveies mention the production of a scope of antimicrobic substances by probiotic bacteriums. Among such substances are organic acids, H peroxide and bacteriocins, which can kill other bugs, alter their metamorphosis, and/or cut down their production of toxins ( Rolfe, 2000 ) .
Bettering growing rate
In carnal nutrition micro-organisms used as probiotics were linked with a proved efficaciousness on the intestine microflora. Administration of probiotic strains individually and in combination was significantly improved provender consumption, feed transition rate, day-to-day addition and entire organic structure weight in animate beings ( Casey et al. , 2007 ) . Probiotics have a positive consequence on assorted digestive procedures, particularly cellulolysis and synthesis of microbic protein ( Yoon and Stern, 1995 ) . Probiotics were stabilizers of ruminal pH and Lactaid, increase the soaking up of some foods and displayed a growth-promoting consequence that was comparable to avilamycin intervention ( Mountzouris et al. , 2007 ) .
Saccharomyces cerevisiae supplementation was associated with an increased flow of microbic protein go forthing the first stomachs and enhanced supply of amino acids come ining the little bowel ( Erasmus et al. , 1992 ) . Putnam et Al. ( 1997 ) , nevertheless observed no consequence with barm on the transition of nitrogen fraction and amino-acids to the little bowel. Amber et Al. ( 2004 ) , working with Lact-A-Bac ( Lactobacillus acidophilus ) , got betterments in the digestiblenesss of energy and of most analytical fractions ( DM, CP, EE ) , including petroleum fiber.
The literature reveals that Saccharomyces cerevisiae does non hold an obvious consequence on entire VFA concentrations, methane production and ammonium hydroxide content ( Chaucheyras et al. , 1997 ) . The consequence of barm supplementation on pH stabilization seems to be strongly dependent on the type of diet tested. The research in stear of Ghorbani et al. , 2002 showed that probiotics do non impact to VFA concentration, but have consequence in ruminal NH3-N value. Proboitics inproved first stomachs acetate concentration in the Equus caballus and swine ( Swyers et al. , 2008 ; Shen et al. , 2009 ) .
Probiotics enhanced animate beings diarrhea by protection against pathogens: The autochthonal enteric bacterial inhibit pathogens by competition to colonisation sites and nutritionary beginning and production of toxic or stimulation of the immune system ( Parvez et al. , 2006 ) . Probiotics can significantly protect hogs against diarrhoea ( Corr et al. , 2007 ) . Probiotics have been shown to be involved in protection against a assortment of pathogens in poulet including Escherichia coli ( Chateau et al. , 1993 ) , Salmonella and Campylobacter ( Stern et al. , 2001 ) , Clostridium and Eimeria ( Dalloul and Lillehoj, 2005 ) . Probiotic activity was mostly repressive since the probiotics bacteriums can cut down the degree of Escherichia coli O157 passenger car and fecal sloughing in cowss and calves ( Brachears et al. , 2003 ) and decreased the badness and continuance of diarrhoea in Escherichia coli O157: H7-infected baby coneies ( Casey et al. , 2007 ) . Probiotic was reduced stomachic redness and bacterial colonisation in Helicobacter pylogy-infected animate beings ( Johnson-Henry et al. , 2005 ) . Probiotics tended to better the wellness position and birthrate of sows ( Alexopoulos et al. , 2004 ) . Probiotic can change the balance of gastrointestinalmicroflora in healthy cats ( Marshall-Jones et al. , 2006 ) and shown to be effectual in forestalling antibiotic associated diarrhoea ( Hawrelak et al. , 2005 ) .
Balance of the intestine microbiota
The coney is both a monogastric and an herbivore. Its digestive system is adapted to agitation of vegetable in colon and degree Celsius & A ; aelig ; semen. In immature coneies, the intestine microflora is non wholly stabilised until 30-40 yearss of age ( Fortun-Lamothe and Boullier, 2007 ) .
The coneies facultative anaerobiotic bacteriums, chiefly Streptoccus sp. and Escherichia coli, reached a maximal degree at the 2nd or 3rd hebdomad of life and so decreased to be residuary or absent after ablactating. The epithelial cells, intestine associated lym-phoid tissue ( GALT ) and commensal vegetations interact with each other organizing a dynamic but delicate equilibrium. A balanced system is necessary to guarantee both alimentary soaking up and protection against pathogens. .
Application of probiotics in farm animal
All stress factors of the ablactation and post-weaning periods can take to increased susceptibleness to gut upsets, infections and diarrhoea ( Modesto et al. , 2009 ) . An interdisciplinary research survey on the manners of action of probiotics in swine, showed that E. faecium NCIMB 10415 reduced the infective bacterial burden of healthy piggies and sows ( Lodemann, et al. , 2006 ) . Enterococcus faecium was besides found to cut down in the colon of weaned pig population of Enterococcus faecalis which is responsible for the oncoming of some instances of post-weaning diarrhea ( Vahjen et al. , 2007 ) . Supplement of the diet of neonatal hogs with a strain of Lactobacillus plantarum resulted in an addition in entire gut populations of lactobacilli in weaned hogs ( Takahashi et al. , 2007 ) . An improved day-to-day weight addition was besides observed in the animate beings that received probiotic Lactobacillus sobrius compared to the control fed group ( Konstantinov et al. , 2008 ) . B. animalis subsp. lactis affected positively the growing public presentation in ablactating piggies and the ratio of bii¬?dobacteria to E. coli in the intestine ( Modesto et al. , 2009 ) .
Kalavathy et al. , 2003 found that a supplementation of 12 Lactobacillus strains in broiler diets improved the bodyweight addition, feed transition rate and was effectual in cut downing abdominal fat deposition. Lactobacilli were besides successful in diminishing mortality due to necrotic enteritis from 60 % to 30 % in a challenge test, when they were given orally at twenty-four hours 1 of life ( Hofacre et al. , 2003 ) . Mountzouris et al. , 2007 indicated that probiotic civilizations modulated the composing and the enzymatic activities of the poulets caecal microi¬‚ora, ensuing in a signii¬?cant probiotic consequence. Probiotics addition egg production and quality ( Panda et al. , 2008 ) . Vila et Al. ( 2009 ) showed that Lactobacillus-based probiotic civilizations signii¬?cantly reduced Salmonella enteritidis recovery in challenged neonatal broiler biddies.
In immature calves, integrating unrecorded barms into the grain, reduced the figure of yearss with diarrhea ( Galvao et al. , 2005 ) . With respect to carnal public presentation, improved weight addition and first stomachs development have been reported in immature calves with several bacterial and yeast strains supplementation ( Adams et al. , 2008 ) . In dairy ruminants, live barms have been shown to better public presentation, the most consistent effects being an addition in dry affair consumption and milk production ( Stella et al. , 2007 ) . A recent meta-analysis survey conducted by Desnoyers et Al. ( 2009 ) showed that barm supplementation, in ruminants, increased dry milk consumption, milk output, first stomachs pH, first stomachs volatile fatty acids concentration, and organic affair digestibleness. Other benei¬?ts have been related to greater entire culturable ruminal bacterial population densenesss and cellulolytic micro-organisms ( Chaucheyras-Durand et al. , 2008 ) and increase i¬?ber digestibleness ( Marden et al. , 2008 ) .
The farming of coneies in intensive system can do physiological and environmental emphasis, lending to the development of digestive jobs and attendant decrease of public presentation. There are of course fewer surveies with probiotics in coneies than in other monogastric farm species. Several surveies exist however, which are limited to the appraisal of the consequence on growing, feed transition, reproduction and mortality ; sometimes cecal activity and digestibleness are studied excessively ( Fal & A ; ccedil ; ao-e-Cunha et al. , 2007 ) .
Bacillus Cereus volt-ampere. toyoi ( Toyocerin® ) , has been found to better the public presentation and/or wellness of turning coneies ( Trocino et al. , 2005 ) . However, seldom survey has been performed to measure the part of the dietetic inclusion of this merchandise either on cecal agitation of turning coneies in the last decennary, or on the alimentary digestibleness and fecal N elimination ( Pascual et al. , 2008 ) . In the test of Yamani et Al. ( 1992 ) , Lacto-Sacc ( a complex merchandise incorporating micro-organisms % Lactobacillus acidophilus, Streptococcus faecium and barms % but besides enzyme activities % peptidase, cellulases, amylase ) improved rough fibre digestibleness at 8 and 12 hebdomads.
Since probiotics can act upon gut microbiology, several writers looked at their effects upon the cecum microbiota, either by numbering bacteriums ( Amber et al. , 2004 ) or their merchandises, VFA in peculiar ( Maertens et al. , 1994 ) . In the survey of Amber et Al. ( 2004 ) , the probiotic significantly increased cellulolytic bacterium counts ( cfu/ml ) . In this survey, cecal pH was unaffected by the probiotic.
MATERIALS AND METHODS
Experimental protocols will be approved by Chulalongkorn University, Thailand.
Model is followed by the figure 1:
6. Statitical analysis and compose the thesis
2. Collect animate beings
3. Conduct experiment
4. Roll up the samples
5. Analyze the samples
7. Register for conference or diary
8. Defend the thesis
Figure 1: The model of the experiment
Animals and experimental design
A sum of 72 coneies ( Newzealand white ) weaned at 28 vitamin D of age are indiscriminately assigned to 4 interventions: 1 ) basal diet ( BD ) ( without antibiotic or probiotic ) , 2 ) low degree of probiotic ( LP ) , 3 ) middle probiotic ( MP ) and 4 ) high degree of probiotic ( HP ) . Each intervention has 3 reproductions with 6 coneies per experimental unit ( equilibrate sex ) . At ablactating, rabbits ase chosen from among those born the same twenty-four hours in the farm from multiparous does, travel from the maternal sector to the fattening sector and set in the coops. The six coneies in each experimental unit come from different litters and had similar unrecorded weight.
Table 2: Ingredient and chemical composing of the basal diets
Ingredient composing, g/kgDM
Land Paddy rice
Sodium chloride ( NaCL )
Calcium oxide ( CaO )
Content ( g/kg natural footing )
DE, MJ/kg DM
ME, MJ/kg DM
Methionine + Cysteine
DE: digestive energy, ME: metabolizable energy, NDF: impersonal detergent fiber, ADF: acerb detergent fiber, CF: petroleum fiber, EE: quintessence infusion, CP: petroleum protein.
During the 8-wk eating period, all coneies are housed in a temperature-controlled baby’s room room ( 25 to 27 & A ; deg ; C ) . Feed and H2O are provided ad libitum. The diets are similar to common commercial diets for turning coneies and explicate utilizing commercial natural stuffs presently adopt by Thai provender manufacturers. They do non incorporate antibiotics, additives, growing boosters or coccidiostatics.
Prior to the eperimental period ( before ablactating ) , both suckling coneies and their female parents will be given ad libitum entree to a commercial provender. From 28 ( ablactating ) to 84 yearss of age coneies are offered the experimental diets.
At the terminal of experiment, two coneies per each experimental unit will be killed by a deadly injection of Na Nembutal to obtain digesta samples in the cecum. The representative digesta from fresh cecum are determine pH instantly, gently squeezed into a preweighed tubing and so shop in a deep-freeze at ?80 & A ; deg ; C until analysis for VFA and N-NH3 concentrations.
At ablactation, the coneies are given designation Markss on the ear, put into bicellular coops and fed the experimental diets. Individual unrecorded weight and coop provender consumption are recorded hebdomadal. Mortality is controlled daily throughout the experimental period. Daily feed consumptions are calculated taking into history the effectual figure of animate beings per coop per twenty-four hours, therefore excepting the consumption of dead animate beings.
Morbidity and mortality
In the computation of morbidity, the ailment coneies are counted merely one time, independently of the continuance of unwellness. The dead animate beings are non considered in the morbidity computation. The wellness hazard is calculated as the amount of morbidity and mortality ( Bennegadi et al. , 2000 ) .
Feed consumptions and growing public presentation
Feed consumptions and BW will be measured at the beginning and terminal of the experiment to find day-to-day feed consumption ( DFI ) , mean day-to-day addition ( ADG ) , and feed transition ratio ( FCR ) :
Dairy feed intake = feed offer – provender garbage ;
Average day-to-day addition = ( weight in initial – weight in concluding ) /number of yearss ;
FCR = DFI / ADG.
Chemical analysis of provender and fecal matters
Feed samples are collected at the start of experiment. Feed and faecal samples are dried in an oven ( 65 & A ; deg ; C ) and land to go through through a 1-mm screen. Each sample is analyzed for DM, OM, CP ( N – 6.25 ) harmonizing to the standard methods of AOAC, 2000 ; NDF and ADF ( Van soest et al. , 1991 ) , Gross energy ( GE ) is determined by an automatic adiabatic O bomb calorimeter ( Parr 1281 Automatic Energy Analyzer, Moline, IL ) .
The experimental design of digestibleness is similar to that of the feeding period, nevertheless, the 10-week old coneies are used to find the fecal evident digestibleness of ME, DM, OM, CP, NDF and ADF ; keeping energy ( Toschi et al. , 2004 ) . Following a 7-day version harmonizing to the method of G & A ; oacute ; mez-Conde et Al. ( 2006 ) . Dry affair consumption and entire fecal end product are recorded daily for each experimental unit over a 5-day aggregation period. On each twenty-four hours, about 50 g of faecal sample is collected from each coop into unfertile plastic bottles and instantly shop at ?20 & A ; deg ; C until farther analysis for alimentary content. Fecal matters produce day-to-day are collected in labelled polythene bags and shop at ?20 & A ; deg ; C. Urine is besides collected for N analysis to cipher the N keeping and find purine derived functions ( PD ) .
Apparent digestibleness ( % )
Food in provender – Food in fecal matters
Food in provender
Retention N = Nitrogen in provender – ( Nitrogen in fecal matters + Nitrogen in piss )
Retention energy ( RE ) = ( 0.58 MEI – 252.4 ) , where Japanese apricot: Metabolizable energy consumption ( Toschi et al. , 2004 ) .
Determine urinary elimination of purine derived functions ( PD )
Merely allantoin and uric acid, non xanthine and hypoxanthine, are present in the coneies urine. PD = Allantoin + acid uric ( Balcells et al. , 1998 ) .
Allantoin is determined by method of Young and Conway ( 1942 ) ,
Acid ucid is analysed harmonizing to the method of Fujihara ( 1987 ) .
Volatile fatty acerb concentration in digesta from the cecum is determined by a gas chromatographic method following the processs of Franklin et Al. ( 2002 ) . About 3 g of cecal chyme are homogenized with 4.5 milliliters metaphosphoric acid ( 4.16 % ) , so centrifuged at 10,000 tens g for 10 min and filtered. 2-ethyl-butyrate ( FLUKA Chemie GmbH, Buchs, Switzerland ) is used as the internal criterion. Parameters: Nukol 30 m x 0.25 millimeters x 0.25 µm capillar column ( Supelco, Bellefonte, PA, USA ) , FID sensor, 1:50 Split ratio, 1 µl injected volume, He 0.84 ml/min. Parameters of the sensor: air 400 ml/min, H 47 ml/min, temperature: injector 250 & A ; deg ; C, sensor 250 & A ; deg ; C, column 150 & A ; deg ; C.
Amonia ( N-NH3 ) concentration in the cecum will be determined by utilizing Kjeldahn method ( AOAC, 2000 ) .
The pH value of the fresh cecal content was determined by a manual automatic pH metre ( OP-110, Radelkis, Hungary ) .
Harmonizing to the literature ( Gouet and Fonty, 1973 ) the cecal microflora in coneies is found to dwell of simple, not sporulated, purely anaerobiotic, Gram-negative Bacteroides. For the microbiological scrutinies of the cecal contents, purely anaerobiotic bacteriums are cultured on Schaedler ‘s agar ( Sharlan Chemie, Barcelona, Spain ) , the selectivity of which was increased by the add-on of esculin ( Merck, Darmstadt, Germany ) , neomycin ( Merck, Darmstadt, Germany ) and Fe-ammonium citrate ( Sharlan Chemie, Barcelona, Spain ) . Subsequently the samples are incubated in anaerobiotic conditions at 37 & A ; deg ; C for 96 hours. Coliforms are cultured on a Chromocult distinction medium ( Merck, Darmstadt, Germany ) . The samples are incubated at 37 & A ; deg ; C, under aerophilic conditions, for 24 hours. Entire aerobe source count is determined on blood agar after incubation at 37 & A ; deg ; C, under aerophilic conditions, for 48 hours. After the incubation clip had elapsed, the settlements are counted with a Titriplaque settlement counter ( LMIM, Esztergom, Hungary ) . The settlement counts are expressed in log10 settlement organizing units ( CFU ) related to 1 g of sample.
Statistical analysis of the informations obtained are carried out by the SAS ( SAS Inst. Inc. , Cary, NC ) utilizing the version 9.0 ( 2002 ) . Consequence of interventions are analyzed by the analysis of discrepancy ( ANOVA ) . The significance of differences between interventions will be tested by the Tukey mothod at ? = 0.05.
The undermentioned theoretical account will be used:
yij = µ + ?i + ?ij I = 1, 2, … ; J = 1, 2, … , ni
yij = an observation J in intervention I
µ = the overall mean
?i = the consequence of intervention I
?ij = random mistake with average 0 and discrepancy ?2
The Chi-Square Test ( SAS 9.0 ) will be used to compare the mortality and morbidity between the interventions
DURATION OF STUDY
Report and paper readying
Topographic points OF STUDY
The experiment will be conducted in experimental farm of Chulalongkorn university.
The equipments of the experiment include digital balances, balance metres, pH metre, nylon bags, plastic tins, fictile cyberspaces and chemical includes acerb sulphuric solution.
Granted fund is supported by Chulalongkorn University with the item is showed in table 3
Table 3: The item in the budgets
Unit of measurements
Entire monetary value,
Land Paddy rice
Blending and pellet the provender
Vaccine and medical specialty
Chemicals for analysis
Car fuel for transfortation
ADVANTAGES OF STUDY
After this experiment, we will cognize the effects of probiotics in experimental parametric quantities and conclude which degree of probiotics is assuring to utilize for the ablactation coneies.