肝素是現今最為重要的抗凝血和抗血栓藥物之一，但迄今沒有一種藥物能完全取代，肝素為高分子硫酸化多醣化合物，因此難以用化學合成。這導致肝素來源須由動物內臟萃取。肝素萃取法有鹽解法、酶解法、酶解結合法等方法，但萃取過程若無持續監控便有可能導致細菌汙染，如2007年的粘質沙雷氏菌汙染事件，但目前尚無研究針對肝素萃取流程座菌相分析，因此本研究以酶解結合法搭配豬腸和豬肺2種肝素萃取來源內臟，並以總基因體技術來分析萃取時菌相的改變，同時搭配生菌數量變化探討萃取流程對生菌的影響。萃取後生菌數在成品中降至0，但萃取過程中需小心環境微生物的影響，成品也須控制濕度以防潮解後遭微生物入侵。在菌相分析的結果，豬肺樣本擁有最高的生物多樣性，但經萃取後則降為最低，豬腸的生物多樣性也是萃取前大於萃取後，顯示萃取對生物多樣性有極大的影響。以HCA和PCoA的分析來看，豬腸與萃取後的豬肺菌相較為接近，可能的原因是環境菌的入侵所導致的，在後續以菌相探討時，發現所有樣本的前2大菌門都為環境常見的Firmicutes和Proteobacteria菌門，有趣的是萃取前豐富度較高的物種都環境菌種，如Brochothrix屬和Peptostreptococcus屬，但萃取後豐富度較高的菌種都為耐逆境的物種，如Macrococcus屬和Psychrobacter屬，而在未萃取樣本有出現特別的菌門，由此可推斷萃取對於菌相有極大的影響，而在萃取前環境菌相對樣本菌項有較大的影響，但樣本攜帶的菌相也會有部分保留。本研究發現萃取過程能降低微生物的殘留量，也對對微生物群落有顯著影響，未來可深入探討微生物汙染的傳播方式與肝素鈉的純度是否會被微生物群落影響。

Heparin, a crucial anticoagulant and antithrombotic drug, is difficult to synthesize chemically and is therefore extracted from animal tissues. Various extraction methods are used, including salt precipitation, enzymatic digestion, and combined enzymatic digestion, but careful monitoring is necessary to prevent bacterial contamination. This study examined microbial changes during heparin extraction from porcine intestines and lungs using combined enzymatic digestion. Genomic techniques were employed to assess the bacterial community, and bacterial counts were analyzed to understand the extraction process's impact. Microbial counts in the final product were reduced to zero, but controlling environmental microbiota and regulating humidity were important to prevent post-extraction microbial invasion. Porcine lungs exhibited the highest biodiversity, which decreased significantly after extraction. Similarly, the biodiversity of porcine intestines decreased post-extraction, indicating the extraction process's influence on microbial diversity. Hierarchical clustering and principal coordinate analyses revealed a close relationship between the microbial communities of porcine intestines and post-extraction porcine lungs, possibly due to environmental microbial invasion. The dominant phyla observed in all samples were Firmicutes and Proteobacteria, commonly found in the environment. Interestingly, environmental strains were abundant before extraction, while stress-tolerant species dominated after extraction. Non-extracted samples displayed distinct phyla, emphasizing the significant impact of the extraction process on the microbial community. The study highlighted the reduction of microbial residuals and the significant effect of the extraction process on microbial communities. Future investigations can explore microbial contamination transmission routes and assess whether the microbial community influences heparin sodium's purity.

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