Mitsuwan Watcharapong, Saengsawang Phirabhat, Jeenkeawpieam Juthatip, Nissapatorn Veeranoot, Pereira Maria de Lourdes, Kitpipit Warangkana, Thomrongsuwannakij Thotsapol, Poothong Saranporn, Vimon Sasi
Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand.
One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand.
Vet World. 2023 May;16(5):1131-1140. doi: 10.14202/vetworld.2023.1131-1140. Epub 2023 May 30.
Probiotics are beneficial microorganisms for humans and animals. In this study, we developed a microencapsulated probiotic with antibacterial activity against avian pathogenic (APEC).
Alignment of the 16S rRNA sequences of the isolate WU222001 with those deposited in GenBank revealed that the isolate was with 99.6% homology. This bacterium was characterized as a probiotic based on its tolerance toward gastrointestinal tract (GIT) conditions, hydrophobicity, and auto-aggregation. The antibacterial activity of the probiotic's culture supernatant against APEC was investigated using a broth microdilution assay. was microencapsulated using sodium alginate and agar with diameters ranging from 47 to 61 μm. Then, physicochemical characteristics and stability of the microcapsules were determined.
The isolate was characterized as a probiotic based on its resistance to low pH, bile salts, and pancreatin, with relative values of 79.2%, 70.95%, and 90.64%, respectively. Furthermore, the bacterium exhibited 79.56% auto-aggregation and 55.25% hydrophobicity at 24 h. The probiotic's culture supernatant exhibited strong antibacterial activity against clinical APEC isolates with minimum inhibitory concentration and minimum bactericidal concentration of 12.5% and 25% v/v, respectively. Microencapsulation-enhanced bacterial viability in GIT compared to free cells. Moreover, 89.65% of the encapsulated cells were released into the simulated intestinal fluid within 4 h. The viable count in microcapsules was 63.19% after 3 months of storage at 4°C.
The results indicated that the culture supernatant of inhibited the growth of APEC. In addition, microencapsulation extends the viability of under harsh conditions, indicating its potential application in the feed production.
益生菌对人和动物有益。在本研究中,我们开发了一种对禽致病性大肠杆菌(APEC)具有抗菌活性的微胶囊化益生菌。
将分离株WU222001的16S rRNA序列与GenBank中保存的序列进行比对,结果显示该分离株的同源性为99.6%。基于其对胃肠道(GIT)条件的耐受性、疏水性和自聚集性,该细菌被鉴定为益生菌。使用肉汤微量稀释法研究了益生菌培养上清液对APEC的抗菌活性。使用海藻酸钠和琼脂对其进行微胶囊化,直径范围为47至61μm。然后,测定了微胶囊的理化特性和稳定性。
基于其对低pH值、胆盐和胰蛋白酶的抗性,该分离株被鉴定为益生菌,相对值分别为79.2%、70.95%和90.64%。此外,该细菌在24小时时表现出79.56%的自聚集性和55.25%的疏水性。益生菌培养上清液对临床APEC分离株表现出较强的抗菌活性,最低抑菌浓度和最低杀菌浓度分别为12.5%和25%(v/v)。与游离细胞相比,微胶囊化提高了细菌在胃肠道中的活力。此外,89.65%的包囊化细胞在4小时内释放到模拟肠液中。在4°C下储存3个月后,微胶囊中的活菌数为63.19%。
结果表明,该菌的培养上清液可抑制APEC的生长。此外,微胶囊化可在恶劣条件下延长该菌的活力,表明其在饲料生产中的潜在应用价值。