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黒毛和種牛の肥育時期による第一胃液性状と細菌叢構成および第一胃粘膜上皮の遺伝子発現に関する研究

尾形, 透 岐阜大学

2020.09.18

概要

黒毛和種肥育牛では,増体や肉質の向上を目的として導入時から長期間にわたって濃厚飼料が多給され,その結果,第一胃発酵が亢進し,第一胃液 pH の低下により亜急性第一胃アシドーシス(SARA)や代謝性疾患が発生している。乳牛では第一胃液 pH の変化に伴って細菌叢構成が変動し,第一胃粘膜上皮における遺伝子発現も変化することが知られている。しかし,黒毛和種肥育牛では第一胃の細菌叢構成や粘膜上皮の遺伝子発現について不明な点が多い。そのため,本研究は黒毛和種牛の肥育時期による第一胃液の性状と細菌叢構成および第一胃粘膜上皮の遺伝子発現の変化を明らかにする目的で実施された。

第1章では,黒毛和種牛の肥育時期による第一胃液性状と細菌叢構成の変化を明らかにする目的で,通常管理下の黒毛和種去勢牛 9 頭を対象として肥育前期 (10-14 カ月齢),中期 (15–22 カ月齢) および後期 (23-30 カ月齢) に第一胃液pH を連続測定,フィステル孔より第一胃液を採材して各種性状と次世代シークエンス法による細菌叢解析が行われた。その結果,第一胃液 pH は肥育時期の進行に伴い有意に低下,リポポリサッカライド (LPS)は中期と後期に前期に比べて有意な高値を,また,Firmicutes 門の構成比は中期に前期に比べて有意な低値を示すことを明らかにした。これらの結果は,黒毛和種牛の肥育中期や後期には第一胃液pH の低下,LPS の産生増加,セルロース分解菌の減少など濃厚飼料多給に伴い第一胃内環境が大きく変化することを示唆している。

第2章では,黒毛和種牛の肥育時期による第一胃液と第二胃液の性状および部位別の細菌叢構成を比較する目的で,第1章と同じ供試牛を用い,pH と各種性状のほか第一胃液,第一胃食渣および第二胃液の細菌叢解析が行われた。その結果,第一胃液 pH は第二胃液 pH に比べて前期と中期に有意な低値,後期には有意な高値を示し,細菌叢構成は第一胃液と第二胃液は類似していたが,第一胃食渣では第一胃液や第二胃液と異なる傾向を示すことを明らかにした。肥育後期に第一胃液pH が第二胃液 pH に比べて高値を示したことは,第一胃粘膜上皮からの重炭酸イオンの供給増加を示唆する重要な知見と思われる。これらのことは,黒毛和種牛の肥育後期には,長期的な濃厚飼料多給による第一胃液 pH の低下に対して,第一胃が緩衝作用を促進して適応していることを示唆している。

第3章では,肥育中期と後期の黒毛和種牛において,濃厚飼料増給に第一胃発酵が適応できるか否かを明らかにする目的で,各 3 頭を対象とし,通常の飼料を給与した対照区 (CON 区)と濃厚飼料割合を約 10%増加した濃厚飼料多給区 (HC 区) に区分して,第一胃液の pH,各種性状および細菌叢の解析が行われた。その結果,第一胃液pH は中期では HC 区で CON 区に比べて有意な低値を示し,後期では両区とも著しい低値を呈して有意差がみられず,揮発性脂肪酸 (VFA) および乳酸濃度はHC 区でCON 区に比べて有意な高値を示した。これらの結果は,黒毛和種牛の肥育後期には濃厚飼料割合の増加による第一胃の発酵と pH変動が良く制御されていることを示唆している。

第4章では,黒毛和種牛の肥育時期による第一胃粘膜上皮の遺伝子発現の変化を明らかにする目的で,第1章と同じ供試牛を用い,前期,中期および後期にフィステル孔を介し第一胃粘膜上皮を採取し,マイクロアレイ法により網羅的遺伝子解析が行われた。その結果,輸送関連遺伝子である SLC family 遺伝子は,中期に前期に比べて down-regulated (11/13),後期に中期に比べ up-regulated(12/19)と判定されるものが多く,VFA 輸送体および SCFA-/HCO -交換輸送体である SLC26A3 遺伝子の発現量は,中期に前期に比べて有意に低下し,後期には中期に比べて有意に増加することを明らかにした。これらの結果は,黒毛和種牛の肥育後期には第一胃粘膜上皮からのVFA 吸収と第一胃内への重炭酸イオン分 泌が促進され,長期間の濃厚飼料多給に適応している可能性を示唆している。

以上のように,本研究により,黒毛和種牛では肥育時期の進行に伴い第一胃液 pH と VFA濃度が低下し,多くのセルロース分解菌を含む Firmicutes 門の構成比が低下することが明らかになった。また,後期には中期に比べて SLC26A3 など VFA や重炭酸イオンの輸送に関連する遺伝子の発現量が増加することから,肥育後期には第一胃粘膜上皮から第一胃内 への重炭酸イオン供給が増加し,この緩衝作用により濃厚飼料多給に順応して恒常性を維持している可能性のあることが示唆された。本研究によって得られた所見は,黒毛和種肥育牛における健康の維持と代謝性疾患の予防に寄与すると考えられた。

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