The objectives of this study were to screen probiotic characteristics of lactobacilli isolated from traditionally homemade koumiss products in Xinjiang and Inner Mongolia of China, and to determine and compare the effect of the Lactobacillus strains in vivo on lipid metabolism in rats fed with a high-lipid diet. Three out of 68 strains of Lactobacillus isolated from the koumiss were screened for bile-salt resistance, acid tolerance, and different hypocholesterolemic properties. The cholesterol-lowering effects of the 3 screened strains were estimated in rats fed a high-lipid diet by determination of serum lipids, liver, and fecal cholesterol, fecal total bile acids, and short-chain fatty acids. After a 4-wk feeding period, in comparison with the control group, the groups LIP-1 and MG9-2 had a significant reduction in serum total cholesterol, triglyceride and low-density lipoprotein cholesterol, and significantly increased high-density lipoprotein, the group E7301 had similar effects on serum lipids, but the change was not significant. The groups LIP-1, MG9-2 and E7301 had a significant reduction in liver cholesterol content and an increase in fecal cholesterol content compared with the control group. Total bile acid excretion was significantly higher in rats fed MG9-2 than the other groups. Rats fed diets containing lactic acid bacteria strains had significantly higher propionic acid and butyric acid concentrations in the feces compared with the control. Results indicated that the 3 screened Lactobacillus strains were able to lower cholesterol in vitro, and reduce cholesterol effectively in vivo. The mechanisms behind the hypocholesterolemic effect of 3 strains are likely to be diverse and will need further investigation.
Angiotensin I-converting enzyme (ACE) inhibitory activities in untreated koumiss and koumiss digested with ACE, pepsin, trypsinase, and chymotrypsin were compared and analyzed. Four novel ACE inhibitory peptides (P(I), P(K), P(M), and P(P)) were purified using ultrafiltration and high performance liquid chromatography (HPLC). The classification study showed that these 4 peptides were of the true inhibitor type. The amino acid sequences of these peptides are YQDPRLGPTGELDPATQPIVAVHNPVIV, PKDLREN, LLLAHLL, and NHRNRMMDHVH, respectively. Their individual IC(50) (50% inhibitory concentration) values were as follows: 14.53+/-0.21microM, 9.82+/-0.37microM, 5.19+/-0.18microM, and 13.42+/-0.17microM. From sequence analysis, we determined that P(I) was part of beta-casein in mare's milk. The 3 peptides P(K), P(M), and P(P) did not correspond with any known milk protein. The results suggest that koumiss is rich in ACE inhibitory peptides, and the ACE inhibitors in koumiss are of the pro-drug type or a mixture of the pro-drug type and the true inhibitor type. These results may provide evidence about the beneficial effects of koumiss, especially on cardiovascular health.
Bacteriophage infection of lactic acid bacteria is considered an important problem worldwide in the food fermentation industry, as it may produce low quality or unsafe foods, cause fermentation failure, and result in economic losses. To increase current knowledge on the properties of Lactobacillus virulent phages, we evaluated the effect of divalent cations, temperature, pH, and chloramphenicol on the adsorption ability of Lactobacillus virulent phage P1. Phage P1 was isolated from the abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The results showed that this phage belonged to the Siphoviridae family. The latent period of this phage was 45min, and the burst time was 90min. Burst size was 132.88±2.37 phage counts expressed per milliliter per infective center. This phage showed good tolerance at different temperatures, but incubation at 50°C only affected its adsorption. Adsorption rate reached a maximum value between 30 and 42°C. A high adsorption value of phage infectivity was obtained from pH 6 to 8. Moreover, calcium ions promoted and increased the adsorption capacity of phage P1, but magnesium ions had negative effects. Chloramphenicol had no effect on phage adsorption. This study increased current knowledge on the characterization and biological aspects of Lactobacillus virulent phages, and may provide some basic information that can be used to design successful antiphage strategies in the food industry.
Bacteriophage infection of lactic acid bacteria is considered one of the biggest worldwide problems in the food industry. Bacteriophages may cause negative effects on the fermentation of various dairy-based products. A virulent bacteriophage was isolated from an abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The characterization and influence of temperature, pH, divalent cations, and chloramphenicol on the adsorption ability of this phage were evaluated. The results showed that this phage belonged to the Siphoviridae family. It exhibited a burst time of 135 min and a burst size of approximately 215 counts expressed per milliliter per infective center. No significant effect was shown to influence its viability and adsorption at 10 to 37°C. More than 90% of phages exhibited infectivity from pH 5 to 9. Divalent ions and chloramphenicol did not have a significant influence on the adsorption of this phage. The information obtained in this study will enrich the database of lactobacilli virulent phages and provide a basis of information for the control of phages in the food fermentation industry.
The effect of thermal treatments and several biocides on the viability of Lactobacillus virulent phage P1 was evaluated. Times to achieve 99% inactivation (T) of phage at different treatment conditions were calculated. The thermal treatments applied were 63, 72, and 90°C in 3 suspension media (de Man, Rogosa, Sharpe broth, reconstituted skim milk, and Tris magnesium gelatin buffer). Phage P1 was completely inactivated in 5 and 10 min at 90 and 72°C, respectively; however, reconstituted skim milk provided better thermal protection at 63°C. When phage P1 was treated with various biocides, 800 mg/L of sodium hypochlorite was required for total inactivation (∼7.3 log reduction) within 60 min, whereas treatment with 100% ethanol resulted in only a ∼4.7 log reduction, and 100% isopropanol resulted in a 5.2-log reduction. Peracetic acid (peroxyacetic acid) at the highest concentration used (0.45%) resulted in only a ∼4.-log reduction of phage within 60 min. The results of this study provide additional information on effective treatments for the eradication of potential phage infections in dairy plants.
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