The newborn calf The newborn calf Probiotic product from Salem, - PDF document

2/20/2011 The Effect of Continuous Low-level Feeding of Aureomycin on Dairy Cows CE Polan , GA McLaren, AH Rakes… - 1962 - West Virginia University, Agricultural … The effect of continuous feeding of chlortetracycline on lactating dairy cows. CE Polan - 1960 - West Virginia University Biohydrogenation of unsaturated fatty acids by rumen microorganisms. CE Polan - 1963 - North Carolina State College. Probiotics and the health of Probiotics and the health of newborn calves? newborn calves? ROLE OF BACTERIA IN ROLE OF BACTERIA IN RUMINANT NUTRITION RUMINANT NUTRITION Bob James Dept. of Dairy Science 1 The newborn calf The newborn calf Probiotic product from Salem, Probiotic product from Salem, VA? VA? • Calf is sterile at birth • Lactobacilli grown on wheat bran • Early microbial colonization - rather • Dose calf with priobiotic in shipped in haphazard? calves housed at Vet Science • Risk of colibacillosis? Early colonization • Challenge with enteropathogenic strain by E. coli. of E. coli. • Can dosing calf with “good”bacteria” colonize intestine and prevent establishment of E. coli. . 3 4 1

2/20/2011 Source of meaningful probiotic? Source of meaningful probiotic? Problems with probiotics – Problems with probiotics – according to W. E. C. Moore – VPI according to W. E. C. Moore – VPI Anerobe Lab Anerobe Lab • Source of organisms for probiotic? • Most intestinal organisms are anaerobes • Older milk-fed calf? not aerobes • Duodenal cannulated • Microflora is a mixed culture of • Collect and mix with whey solution organisms. • Intestinal fluid is of low palatability. • Liklihood of single species or few species to successfully populate is nill • Intestine is rapidly populated by organisms at birth. 5 6 Influence of “probiotic” on resistance to Influence of “probiotic” on resistance to Results Results enteropathogenic E. coli challenge enteropathogenic E. coli challenge James et al, 1976 James et al, 1976 • Calves receiving probiotic inoculum had less diarrhea and higher ADG for 1 st 14 • Calves entered our facility @~ 3 hour of days. age • Colostrum fed @ ~ 5 hours of age • No E.coli challenge • E. coli challenge @ 12 hours • E. coli challenge @ 24 hours • All with or without “probiotic” inoculum of 200 ml of duodenal fluid from milk-fed calf. • Whey solution as carrier 7 8 2

2/20/2011 Serum gamma globulin – g/100 ml @ 24 h Serum gamma globulin – g/100 ml @ 24 h Follow up study Follow up study James et al., (1978) James et al., (1978) Inoculum O 12 h 24 h Means • Three Treatments challenge challenge • Colostrum @ 2.5 hours of age No 1.1 0.94 1.68 1.24 • Colostrum and inoculum (200 ml duodenal inoculum fluid) concurrently @ 2.3 hours of age Inoculum .5 0.49 .59 .53 • Inoculum followed three h later by colostrum Means .80 .72 1.14 .89 @ 5.6 hours of age • No E. coli challenge 9 10 Mean total protein and gamma globulin (g/dl) @ Mean total protein and gamma globulin (g/dl) @ Cause of apparent depression Cause of apparent depression 24 hours of age. 24 hours of age. in Ig absorption? in Ig absorption? Treatment Total serum Gamma globulin • Live bacteria? protein • Cell walls of bacteria (endotoxins)? No inoculum 5.97+/-0.94 1.05+/-.36 • Carrier for the inoculum – dried whey? Colostrum and 6.07+/-0.64 1.08+/-.41 inoculum concurrent Delayed 5.22+/-.59 .76+/-.31 colostrum Delayed colostrum calves had lower protein and gamma globulin 11 12 3

2/20/2011 Intensive study Intensive study Treatments Treatments James et al, (1981) James et al, (1981) • Utilized “gut loops” in newborn calves ~ • Live bacteria – .3ml duodenal fluid – 9 ml 8.6 hours at beginning of treatments anaerobic rumen fluid glucose cellobiose broth (RGC). Anaerobic culture • Constructed beginning 1.8 M anterior to ileocecal junction • Autoclaved culture • Each calf received all treatments • Sterile RGC broth • Incubated for 4 hours • Inject loops with 125 I – gamma globulin in electrolyte solution incubated for 75 min. 13 14 Treatment means for uptake, bacterial Treatment means for uptake, bacterial Three Q’s of colostrum management Three Q’s of colostrum management no. in tissue and inoculum no. in tissue and inoculum • Quickness – feed as • Not colostrum again! Uptake of gamma globulin a Bacterial growth in tissue b soon as possible Live Sterile Autoclaved Live Sterile Autoclaved bacteria broth bacteria bacteria broth bacteria with a goal of less 2.09 +/- 3.18 +/- 3.56 +/- 594+/- 86 +/- 82 220 than 6 hours 1.15 1.50 1.49 316 • Quantity – 4 liters in 1 st 12 hours a milligrams gamma globulin internalized/g tissue b Bacteria (X10 6 ) • Quality - >50g/liter Uptake reduced only with live bacteria culture – (P<.05) # organisms significantly related to uptake 15 16 4

2/20/2011 Add another letter to the list? Add another letter to the list? Microbial risks associated with Microbial risks associated with feeding colostrum feeding colostrum • “C” for cleanliness 17 18 Review Ig absorption in the calf Review Ig absorption in the calf Cessation of Ig absorption Cessation of Ig absorption Ig Absorption Calf epithelium ~ 24 hours • Cessation termed closure • Large MW macromolecules • Differs by species absorbed from Jejunum and proximal ileum • In the calf • 1 st 2 – 24 hours of life • Not as diet dependent • Variable efficiency • Very variable onset – 6 – 24 hours • Little selectivity in uptake • Precipitous decline in absorptive efficiency • Some selectivity in transfer through serosal side Corley et al. 1977 19 20 5

2/20/2011 Closure - One step process where Closure - One step process where Replacement by generation of Replacement by generation of uptake and transfer ceases uptake and transfer ceases cells incapable of absorption. cells incapable of absorption. • Theories • Micropinocytotic IgG transfer by newborn calf enterocytes • Development of gastric and enzymatic function • Existence of a receptor mediated transport system? • Reduction in permeability of villus epithelial cells • Relationship between apoptosis in • Replacement by generation of cells cessation of Ig transfer incapable of pinocytocis 21 22 Apoptosis and IgG absorption in Apoptosis and IgG absorption in IgG staining in duodenum of IgG staining in duodenum of goats Castro-Alonso et al. (2008) goats Castro-Alonso et al. (2008) goat kids harvested at ….. goat kids harvested at ….. • 10 new born kids • Fed colostrum – 2,000 mg IgG/kg body weight – 2X @ 2 and 14 h • Sacrificed - birth – 60 d of age. • Assessed for apoptotic cells and stained Birth - no IgG immunoreactivity for IgG. Day 1 – IgG internalized Day 2 + IgG was bound to enterocytes but minimal internalization Day 60 – No staining for IgG 23 24 6

2/20/2011 Terminal deoxynucleotidyl transferase nick Terminal deoxynucleotidyl transferase nick Conclusions Conclusions end labeling (TUNEL) – identifies apoptotic end labeling (TUNEL) – identifies apoptotic cells cells • Relationship of IgG absorption with apoptosis in the intestine? • Apoptosis is influenced by composition of colonizing intestinal bacteria in pigs (Willing and Van Kessel, 2007) • How to delay apoptosis? Birth – TUNEL staining at center of villus Day 1 – staining is significantly reduced after day 1 Day 60 – only cells at villus terminus are staining 25 26 Feeding heat-treated colostrum or unheated colostrum Feeding heat-treated colostrum or unheated colostrum IgG (g/L) and SPC of colostrum IgG (g/L) and SPC of colostrum with two different bacterial concentrations to neonatal with two different bacterial concentrations to neonatal calves - Elizonda-Salazar and Heinrichs, 2009 calves - Elizonda-Salazar and Heinrichs, 2009 Serum protein and IgG @24 h Serum protein and IgG @24 h • Treatments Item Low bacteria High bacteria Heat – treated • Colostrum frozen - low bacteria IgG (g/L) 69.55 69.55 66.17 SPC x10 3 /ml 9.332 40.738 .645 • Colostrum pasteurized – 60 o C / 30 min. Serum protein @ 57 56.2 62.5 then frozen 24h (g/L) • Colostrum stored at 20 o C for 24 h then IgG @ 24 h (g/L) 20.2 20.1 26.7 AEA of IgG 35.4 32.4 43.9 frozen @ 24 h (%) • First feeding 3.8 liters / 68g IgG/liter - 1.5 – 2 h after birth with esophageal feeder. Heat treating improved AEA and IgG at 24 h. High bacteria load was relatively low / less than 100,000 goal (McGuirk) 27 28 7

2/20/2011 Role for microbial colonization Role for microbial colonization Intestinal microflora and the Intestinal microflora and the and Ig absorption? and Ig absorption? absorptive surface absorptive surface • Rate of intestinal cell production in the • Colostrum deprived calf receiving E. coli crypts O55 • Migration of cells up the villus and • Exfoliation of desquamation from the tips influenced by microvilli microbial colonization? • Intracellular • Microbes may occupy binding sites on penetration of the apical plasma membrane. epithelial cells Corley et al. , 1977 29 30 How is colostrum managed on How is colostrum managed on many dairies? many dairies? Colostrum deprived Colostrum fed calf 31 32 8