【摘要】：本試驗(yàn)在前期體內(nèi)群體研究的基礎(chǔ)上,選擇2株瘤胃厭氧真菌和3株纖維降解細(xì)菌,進(jìn)行單一培養(yǎng)、混合培養(yǎng)和序列培養(yǎng),研究菌株之間的相互關(guān)系；再以不同碳源進(jìn)行培養(yǎng),明確碳源組成對(duì)瘤胃厭氧真菌和纖維降解細(xì)菌互作關(guān)系的影響；并結(jié)合同位素示蹤技術(shù),以13C標(biāo)記的純纖維素作為菌株培養(yǎng)的發(fā)酵底物,精確追蹤纖維降解的去路問題,更準(zhǔn)確的來(lái)評(píng)價(jià)研究瘤胃厭氧真菌和細(xì)菌在纖維降解中的貢獻(xiàn)及相互作用。試驗(yàn)結(jié)果表明,經(jīng)纖維降解特性的研究發(fā)現(xiàn)拜氏梭菌、糞腸球菌、白色瘤胃球菌、Neocallimastix屬菌和Cyllamyces屬菌纖維降解情況及生長(zhǎng)狀態(tài)良好。拜氏梭菌分別與腸球菌、Neocallimastix屬菌混合培養(yǎng)具有促進(jìn)發(fā)酵的作用,而拜氏梭菌與Cyllamyces屬菌混合培養(yǎng)無(wú)明顯作用,拜氏梭菌與白色瘤胃球菌混合培養(yǎng)出現(xiàn)抑制發(fā)酵的現(xiàn)象；白色瘤胃球菌分別與腸球菌、Neocallimastix屬菌、Cyllamyces屬菌混合培養(yǎng)有抑制作用；腸球菌與Neocallimastix屬菌混合培養(yǎng)有促進(jìn)作用；Cyllamyces屬菌分別與腸球菌、Neocallimastix屬菌混合培養(yǎng)無(wú)明顯的相互作用。進(jìn)一步做序列培養(yǎng)后發(fā)現(xiàn),拜氏梭菌、糞腸球菌與白色瘤胃球菌之間存在相互影響。纖維降解細(xì)菌與厭氧真菌混合培養(yǎng)若為促進(jìn)作用,則是厭氧真菌促進(jìn)了纖維降解細(xì)菌；纖維降解細(xì)菌與厭氧真菌混合培養(yǎng)若為抑制作用,則是纖維降解細(xì)菌抑制了厭氧真菌�；旌吓囵B(yǎng)的促進(jìn)作用主要是由前一株菌的代謝產(chǎn)物引起的,但是干物質(zhì)降解率的增加還是由于纖維結(jié)合位點(diǎn)的作用；混合培養(yǎng)中菌體蛋白濃度、產(chǎn)氣量的抑制作用的原因表現(xiàn)在代謝產(chǎn)物與纖維結(jié)合位點(diǎn),氨氮濃度、TVFA、濾紙酶活力的抑制的根源在纖維結(jié)合位點(diǎn)。經(jīng)過三種不同比例的碳源比較,粗料與精料比為7：3的組合其微生物產(chǎn)量、降解率、VFA和NH3-N濃度等指標(biāo)的結(jié)果較好,反映了此組合利于微生物的生長(zhǎng)及對(duì)底物的發(fā)酵與利用。通過對(duì)底物纖維素添加13C同位素進(jìn)行示蹤可知,單一培養(yǎng)時(shí)纖維素的主要去路為VFA,混合培養(yǎng)時(shí),VFA中的13C豐度急劇減少。在纖維素與淀粉為7：3時(shí),碳的主要去路為VFA,氣體中的碳比例有所增加。
[Abstract]:On the basis of the previous population study in vivo, two strains of rumen anaerobes and three strains of fiber-degradable bacteria were selected for single culture, mixed culture and sequence culture to study the relationship between the strains, and then the effects of carbon source composition on the interaction between rumen anaerobic fungi and fiber-degradable bacteria were determined. Combined with isotopic tracer technique, using 13 C labeled pure cellulose as the fermentation substrate of strain culture, the problem of fiber degradation was accurately traced, and the contribution and interaction between rumen anaerobes and bacteria in fiber degradation were evaluated and studied more accurately. The results showed that Clostridium faecalis, Enterococcus albicans, Neocallimastix and Cyllamyces had good fiber degradation and growth. The mixed culture of Clostridium berghei with Enterococcus and Neocallimastix could promote the fermentation, but the mixed culture of Clostridium Bayer and Cyllamyces had no obvious effect, and the mixed culture of Clostridium Bayer and Clostridium albicans could inhibit the fermentation, and the mixed culture of Clostridium albicans with Enterococcus, Neocallimastix and Cyllamyces could inhibit the fermentation, and the mixed culture of Enterococcus and Neocallimastix could promote the fermentation, and the mixed culture of Clostridium berberis and Clostridium albicans could inhibit the fermentation, inhibit the fermentation of Clostridium albicans and inhibit the fermentation. There was no obvious interaction between Cyllamyces and Enterococcus and Neocallimastix. After further sequence culture, it was found that there was an interaction between Clostridium monocytogenes, Enterococcus faecalis and Enterococcus albicans. If the mixed culture of fiber-degradable bacteria and anaerobes was promoted, it was anaerobes that promoted fiber-degradable bacteria, and if the mixed culture of fiber-degradable bacteria and anaerobes was inhibitory, it was the fiber-degradable bacteria that inhibited anaerobes. The promoting effect of mixed culture was mainly caused by the metabolites of the previous strain, but the increase of dry matter degradation rate was still due to the role of fiber binding sites. The reasons for the inhibition of bacterial protein concentration and gas production in mixed culture were the binding sites of metabolites to fiber, ammonia nitrogen concentration and TVFA, filter paper enzyme activity. Through the comparison of three different proportion of carbon sources, the results of microbial yield, degradation rate, VFA and NH3-N concentration were better when the ratio of crude material to concentrate was 7:3, which reflected that the combination was beneficial to the growth of microorganisms and the fermentation and utilization of substrate. By tracing the addition of 13 C isotopes to the substrate cellulose, it can be seen that the 13C abundance in VFA decreased sharply when the main route of cellulose in single culture was VFA, mixed culture. When the ratio of cellulose to starch is 7: 3, the main route of carbon is the ratio of carbon in VFA, gas.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S852.6

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