66 th EAAP Annul Meeting (Wrsw, 215) Chnges in ph nd microil composition in the ruminl nd reticulr fluids of SARA cttle Shigeru SATO Coopertive Deprtment of Veterinry Medicine, Fculty of Agriculture, Iwte University, Moriok, Iwte, Jpn
A rdio-trnsmission ph-mesurement system 15mm in length, 3mm in dimeter, nd 22g in weight Glss electrode Appernce of the ph sensor ttched metl chin for recovery ph correltion etween continuous recording nd spot smpling Circdin chnges in ruminl ph nd temperture of cttle fed SARA-diet Orl dministrtion of the sensor 7. 6.5 6. 5.5 5. Chnges in ruminl ph of periprturient cow 1 2 3 Weeks fter prturition
Bckground (1) The ph sensor could enter the reticulum or could move from the rumen to reticulum when dministered orlly. The reticulr ph is slightly higher thn the ruminl ph (Kruse et l., 25; Lne et l., 1968) due to mixing nd dilution with sliv (Duffield et l., 2) However, the reltions etween ruminl nd reticulr ph re unknown. Ruminl nd reticulr ph were investigted in cttle with repeted induced SARA to revel the reltions etween the oth ph.
Bckground (2) Severl studies hve een conducted on the rumen microil community in SARA cows (Chen et l., 212; Golder et l., 21; Mo et l., 213; Hernndez-Snri et l., 212). Studies out microil community in SARA cttle re sed on long-term induction, however, the community in cttle with short-term induced SARA is unknown. Bcteril composition in cttle with induced SARA were investigted to revel chnges in composition of the rumen nd reticulum ginst cute nd short-term chnge in feeding.
Mterils nd Methods (1) Cttle exmined: Eight rumen-cnnulted Holstein steers (ge, 8-1 months; weight, 18-2 kg) Feeding nd experimentl clves: Cttle fed hy- or concentrte-rich diet (2% hy nd 8% concentrte) for every 7 dys during th -concecutive times. Hy Conc. Hy Conc. Hy Conc. Hy Conc. Smpling of the ruminl nd reticulr fluids Mesurements: Ruminl nd reticulr ph : A rdio-trnsmission ph-sensor (DKK-To Ymgt, Jpn) ws plced in the ventrl sc of the rumen nd the reticulum, nd ph ws mesured every 1 min.
ph Mterils nd Methods (2) Circdin ph chnge: Cttle ws divided into 3 groups y circdin chnges of the ruminl ph in ll hy- nd concentrte-fed period, nd the ruminl nd reticulr ph were compred every 3 groups. 7.5 7. ph, durtion in dy 6.5 6. SARA (n=8, 57 smple) ph <5.6, 3 hrs< 5.5 5. 8 9 11112131151617181922122232 1 2 3 5 6 7 Time of dy Slight-SARA (n=8, 32 smple) Non-SARA (n=8, 37 smple) ph <5.6, <3 hrs ph <5.6, hr
Mterils nd Methods (3) Bcteril composition: Bcteril composition in the rumen nd reticulum (1 st experiment) were nlyzed y next genertion pyrosequence (16S rrna rrngement nlysis) nd rel time PCR using cteril DNA extrction from the fluids. Fluids Wsh DNA solution Bcteril lysis, DNA extrction nd purifiction 16s rrna rrngement nlysis (Kim et l.,213 ) F-primer Rel time PCR (Bekele et l.,21 ) primer polymerse P V1 V2 R-primer Smple1 OTU1 ;size=2 Smple1 OTU ;size=15 Kingdom 界 Phylum 門 Clss 網 Order 目 Fmily 科 Genus 属 Species 種 Bcteri Firmicutes Clostridi Clostridiles Euctericee Eucterium Bcteri Bcteroidetes Bcteroidi Bcteroidles Prevotellcee Prevotell Eucterium coprostnoligenes rumen cterium NKC17 : : : : : : : :
ph 7. Rumen 第一胃 第二胃 6.5 6. 5.5 Hy Conc. 1 2 3 5 6 7 8 Weeks fter feeding Chnges in 2-hr men ruminl nd reticulr ph of cttle with repeted induced SARA t hy- nd concentrte-fed period. M±SE (n=8), P<.5 (vs. rumen in the sme dy), P<.5 (vs. one dy efore feeding chnge in the sme site)
ph ph ph 7. SARA 7. Slight-SARA 7. Non-SARA 6.5 6. 6.5 6. c d 6.5 6. 5.5 5. 8 11211618222 2 6 Time of dy 5.5 5. Rumen 第一胃第二胃 8 1 12 1 16 18 2 22 2 6 Time of dy 5.5 5. 8 112 1 1618 222 2 6 Time of dy Circdin chnges in 1-hr men ruminl nd reticulr ph of cttle with SARA, slight SARA nd non-sara. M±SE, P<.5 (vs. rumen in the sme time) SARA Slight-SARA Non-SARA Rumen 2-hr men 5.92±.2 6.23±.2 6.±.3 Hours (<ph5.6) 6.89±.6 1.±.15.±. 2-hr men 6.8±.5 6.37±.3 6.7±. Hours (<ph5.6) 5.7±1.5 1.33±.32.6±.28
1% No.1 1% No.2 unclssified Bcteri (miscellneous) その他 others Selenomonus 75% 75% Oricterium Streptococcus 5% 5% Bcteroides cndidte division TM7 25% 25% Acetivirio Pseudoutyrivirio % 1% 75% % Rumen No.3 Rumen 1% hy con 75% No. Olsenell Roseuri unclssified Lchnospircee unclssified Clostridiles (miscellneous) Brnesiell Pimelocter 5% 5% Oscillicter Succiniclsticum 25% 25% Ruminococcus Eucterium % Rumen Rumen Rumen Rumen Hy Concentrte hy con % Rumen Rumen Rumen Rumen Hy hy Concentrte con Butyrivirio Clostridium Prevotell Comprison of cteril community (genus) in the ruminl nd reticulr fluid of cttle (n=) with hy- nd concentrte-fed period t 1 st experiment y 16S rrna gene pyrosequencing.
Numers of cteril phylum, genus nd species in cttle with hy- nd concentrte-fed period t 1 st experiment y 16S rrna gene pyrosequencing Phylum Genus Species Cttle Hy Concentrte Hy Concentrte Hy Concentrte No. Ⅰ Ⅱ Ⅰ Ⅱ Ⅰ Ⅱ Ⅰ Ⅱ Ⅰ Ⅱ Ⅰ Ⅱ 1 16 2 11 9 1 16 97 9 371 16 229 28 2 16 17 7 8 15 13 75 85 325 359 165 223 3 11 15 9 8 132 15 77 8 317 335 177 22 1 13 5 6 97 17 81 68 237 25 358 195 Men 13.3 16.3 8. 7.8 129.5 139.8 82.5 82.8 312.5 31. 232.3 236. SE 1.6 1.5 1.3.6 11.2 11.9 5. 5. 27.8 33.6.2 18.7 Ⅰ: Rumen, Ⅱ:, P<.5 (vs. hy-fed period in the sme site)
M. elsdenii S. dextrinosol vens S. ovis R. flvefcien s R. lus S. ruminntiu m F. succinogen es Totl methnoge n Totl Bcteri 濃厚乾草濃厚乾草濃厚乾草濃厚乾草濃厚乾草濃厚乾草濃厚乾草濃厚乾草濃厚乾草 log 2 6 8 1 12 Totl cteri Hy Rumen Rumen Totl methnogen Hy Rumen Rumen Firocter Hy Rumen Rumen succinogenes Selenomons Hy Rumen Rumen ruminntium Ruminococcus lus Hy Rumen Rumen Ruminococcus Hy Rumen Rumen flvefciens Streptococcus ovis Hy Rumen Rumen Succinivirio Hy Rumen Rumen dextrinosolvens Megspher elsdenii Hy Rumen Rumen Comprison of cteril numers in the ruminl nd reticulr fluids of cttle with hy- nd concentrte-fed period t 1 st experiment y rel time PCR. M±SE (n=8), P<.5, P<.1 (vs. rumen in the sme feeding period), P<.5, P<.1 (vs. hy feeding period in the sme site).
Conclusion Ruminl ph ws lower thn tht of the reticulum, however, significnt positive correltion ws oserved etween the ruminl nd reticulr ph. The reticulr ph could e used to detect SARA cttle, s opposed to using the ruminl ph. Bcteril diversity nd composition were similr in the rumen nd reticulum, nd were lower nd simpler in concentrte-fed period. Chnges in cteril composition of SARA cttle might e relted to the decrese in numer of cteri tht occur following mrked chnges in the ruminl nd reticulr ph.
Butyric cid Butyric cid Butyric cid Propionic cid Propionic cid Propionic cid Acetic cid Acetic cid Acetic cid Totl VFA Totl VFA Totl VFA 2 1 12 SARA Slight-SARA non-sara Rumen 第一胃 第二胃 2 1 8: 1: 2: 12 8: 1: 2: 12 8: 1: 2: 2 1 8 8 8 6 2 8: 1: 2: 8: 1: 2: 6 2 8: 1: 2: 8: 1: 2: 6 2 8: 1: 2: 8: 1: 2: 2 8: 1: 2: 2 8: 1: 2: 2 8: 1: 2: Concentrtion of totl VFA, cetic, propionic nd utyric cid in the ruminl nd reticulr fluid of cttle with SARA, slight SARA nd non-sara. M±SE (n=8), P<.5, P<.1 (vs. rumen in the sme time), P<.5, P<.1 (vs. 8: in the sme site)