Effects of chemical structure, concentration, and pH on antimicrobial activity of conjugated bile acids were investigated in 4 strains of lactobacilli. Considerable differences were observed in the antimicrobial activity between the 6 human conjugated bile acids, including glycocholic acid, taurocholic acid, glycodeoxycholic acid, taurodeoxycholic acid, glycochenodeoxycholic acid, and taurochenodeoxycholic acid. Glycodeoxycholic acid and glycochenodeoxycholic acid generally showed significantly higher antimicrobial activity against the lactobacilli, but glycocholic acid and taurocholic acid exhibited the significantly lower antimicrobial activity. Glycochenodeoxycholic acid was selected for further analysis, and the results showed its antimicrobial activity was concentration-dependent, and there was a significantly negative linear correlation (R2 > 0.98) between bile-antimicrobial index and logarithmic concentration of the bile acid for each strain of lactobacilli. Additionally, the antimicrobial activity of glycochenodeoxycholic acid was also observed to be pH-dependent, and it was significantly enhanced with the decreasing pH, with the result that all the strains of lactobacilli were unable to grow at pH 5.0. In conclusion, chemical structure, concentration, and pH are key factors influencing antimicrobial activity of conjugated bile acids against lactobacilli. This study provides theoretical guidance and technology support for developing a scientific method for evaluating the bile tolerance ability of potentially probiotic strains of lactobacilli.Measuring circulating progesterone (P4) of dairy cows is a key component of many research studies dealing with basic and applied reproduction physiology. The gold standard in dairy cows for the measurement of P4 in serum is radioimmunoassay (RIA), but it generates radioactive waste and requires licensed facilities. The purpose of this study was to develop and validate an in-house competitive enzyme immunoassay (EIA) to measure the P4 concentration in serum of dairy cattle. The secondary objective was to validate a commercial EIA. In the present study, a competitive EIA was developed using commercially available antibodies and conjugates. Ninety-six well microtiter plates were coated with the secondary antibody and incubated overnight. Following a washing step, the wells were blocked using the primary antibody. Serum samples were prepared by first extracting P4 using petroleum ether, then diluted in working conjugate solution. Samples were pipetted into the coated and blocked plates, then the matching HRP conjthe in-house EIA, the intraassay CV were all 0.4 ng/mL, and a perfect agreement with RIA using a threshold of 1.0 ng/mL.Dietary lecithin is a source of choline. Our objective was to evaluate the effects of dietary deoiled soy lecithin feeding on circulating choline, choline metabolites, and the plasma phospholipid profile in lactating dairy cows fed fractionated palm fatty acids. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 d in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate or high palmitic acid content at 1.75% of ration dry matter (moderate and high palmitic acid containing 72 or 99% palmitic acid in fat supplement, respectively; n = 8/palm fat diet). Within each palm fat group, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design with 14-d experimental periods. A 14-d covariate period was used to acclimate cows to palm fat feeding without lecithin supplementation. Blood sampling occurred during the final 3 d of each experianced endogenous phospholipid synthesis.Mastitis is one of the most common diseases in dairy cattle, causing severe economic losses to dairy farmers. Mastitis usually occurs due to intramammary infection (IMI) caused by a variety of pathogenic bacteria. Although good progress has been made in understanding genetics of pathogen-specific clinical mastitis, studies involving genetic analysis of pathogen-specific IMI are scarce. The overall objective of this study was, therefore, to assess genetic variation of overall and pathogen-specific IMI in nonclinical primiparous and multiparous cows using bacterial culture. Data and milk samples were collected over a 2-yr interval as part of the Canadian Bovine Mastitis Research Network. The final data set contained records of 46,900 quarter milk samples from 3,382 clinically healthy primiparous and multiparous Holstein cows from 84 dairy herds. For the genetic analysis, we considered the following 7 traits overall IMI, non-aureus staphylococci (NAS) IMI, contagious pathogen IMI, environmental pathogen IMI, maj (90% decile) according to their IMI trait-specific estimated breeding values, respectively. Pathogen-specific IMI traits and overall IMI had weak to moderate positive genetic correlations [ranging from 0.11 to 0.81 (±0.11 to 0.22)] with SCS. Therefore, selection for lower SCS will improve resistance to IMI. https://www.selleckchem.com/products/crenolanib-cp-868596.html However, based on the observed weak to moderate rank correlations (0.04 to 0.47) between pathogen-specific IMI traits and SCS, selection for lower SCC will not improve resistance to IMI from every pathogen-specific IMI group in the same manner. Therefore, despite low heritability estimates, there was sizeable genetic variation for pathogen-specific IMI traits, indicating that long-term direct genetic selection for pathogen-specific IMI can improve pathogen-specific IMI resistance.There is an increasing demand for dairy products, but the presence of food-spoilage bacteria seriously affects the development of the dairy industry. Bacteriocins are considered to be a potential antibacterial or antibiofilm agent that can be applied as a preservative. In this study, bacteriocin BM173 was successfully expressed in the Escherichia coli expression system and purified by a 2-step method. Furthermore, it exhibited a broad-spectrum antibacterial activity, high thermal stability (121°C, 20 min), and broad pH stability (pH 3-11). Moreover, the minimum inhibitory concentration values of BM173 against E. coli ATCC 25922 and Staphylococcus aureus ATCC 25923 were 14.8 μg/mL and 29.6 μg/mL, respectively. Growth and time-kill curves showed that BM173 exhibited antibacterial and bactericidal activity. The results of scanning electron microscopy and transmission electron microscopy demonstrated that BM173 increased membrane permeability, facilitated pore formation, and even promoted cell lysis. The disruption of cell membrane integrity was further verified by propidium iodide uptake and lactic dehydrogenase release.
Effects of chemical structure, concentration, and pH on antimicrobial activity of conjugated bile acids were investigated in 4 strains of lactobacilli. Considerable differences were observed in the antimicrobial activity between the 6 human conjugated bile acids, including glycocholic acid, taurocholic acid, glycodeoxycholic acid, taurodeoxycholic acid, glycochenodeoxycholic acid, and taurochenodeoxycholic acid. Glycodeoxycholic acid and glycochenodeoxycholic acid generally showed significantly higher antimicrobial activity against the lactobacilli, but glycocholic acid and taurocholic acid exhibited the significantly lower antimicrobial activity. Glycochenodeoxycholic acid was selected for further analysis, and the results showed its antimicrobial activity was concentration-dependent, and there was a significantly negative linear correlation (R2 > 0.98) between bile-antimicrobial index and logarithmic concentration of the bile acid for each strain of lactobacilli. Additionally, the antimicrobial activity of glycochenodeoxycholic acid was also observed to be pH-dependent, and it was significantly enhanced with the decreasing pH, with the result that all the strains of lactobacilli were unable to grow at pH 5.0. In conclusion, chemical structure, concentration, and pH are key factors influencing antimicrobial activity of conjugated bile acids against lactobacilli. This study provides theoretical guidance and technology support for developing a scientific method for evaluating the bile tolerance ability of potentially probiotic strains of lactobacilli.Measuring circulating progesterone (P4) of dairy cows is a key component of many research studies dealing with basic and applied reproduction physiology. The gold standard in dairy cows for the measurement of P4 in serum is radioimmunoassay (RIA), but it generates radioactive waste and requires licensed facilities. The purpose of this study was to develop and validate an in-house competitive enzyme immunoassay (EIA) to measure the P4 concentration in serum of dairy cattle. The secondary objective was to validate a commercial EIA. In the present study, a competitive EIA was developed using commercially available antibodies and conjugates. Ninety-six well microtiter plates were coated with the secondary antibody and incubated overnight. Following a washing step, the wells were blocked using the primary antibody. Serum samples were prepared by first extracting P4 using petroleum ether, then diluted in working conjugate solution. Samples were pipetted into the coated and blocked plates, then the matching HRP conjthe in-house EIA, the intraassay CV were all 0.4 ng/mL, and a perfect agreement with RIA using a threshold of 1.0 ng/mL.Dietary lecithin is a source of choline. Our objective was to evaluate the effects of dietary deoiled soy lecithin feeding on circulating choline, choline metabolites, and the plasma phospholipid profile in lactating dairy cows fed fractionated palm fatty acids. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 d in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate or high palmitic acid content at 1.75% of ration dry matter (moderate and high palmitic acid containing 72 or 99% palmitic acid in fat supplement, respectively; n = 8/palm fat diet). Within each palm fat group, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design with 14-d experimental periods. A 14-d covariate period was used to acclimate cows to palm fat feeding without lecithin supplementation. Blood sampling occurred during the final 3 d of each experianced endogenous phospholipid synthesis.Mastitis is one of the most common diseases in dairy cattle, causing severe economic losses to dairy farmers. Mastitis usually occurs due to intramammary infection (IMI) caused by a variety of pathogenic bacteria. Although good progress has been made in understanding genetics of pathogen-specific clinical mastitis, studies involving genetic analysis of pathogen-specific IMI are scarce. The overall objective of this study was, therefore, to assess genetic variation of overall and pathogen-specific IMI in nonclinical primiparous and multiparous cows using bacterial culture. Data and milk samples were collected over a 2-yr interval as part of the Canadian Bovine Mastitis Research Network. The final data set contained records of 46,900 quarter milk samples from 3,382 clinically healthy primiparous and multiparous Holstein cows from 84 dairy herds. For the genetic analysis, we considered the following 7 traits overall IMI, non-aureus staphylococci (NAS) IMI, contagious pathogen IMI, environmental pathogen IMI, maj (90% decile) according to their IMI trait-specific estimated breeding values, respectively. Pathogen-specific IMI traits and overall IMI had weak to moderate positive genetic correlations [ranging from 0.11 to 0.81 (±0.11 to 0.22)] with SCS. Therefore, selection for lower SCS will improve resistance to IMI. https://www.selleckchem.com/products/crenolanib-cp-868596.html However, based on the observed weak to moderate rank correlations (0.04 to 0.47) between pathogen-specific IMI traits and SCS, selection for lower SCC will not improve resistance to IMI from every pathogen-specific IMI group in the same manner. Therefore, despite low heritability estimates, there was sizeable genetic variation for pathogen-specific IMI traits, indicating that long-term direct genetic selection for pathogen-specific IMI can improve pathogen-specific IMI resistance.There is an increasing demand for dairy products, but the presence of food-spoilage bacteria seriously affects the development of the dairy industry. Bacteriocins are considered to be a potential antibacterial or antibiofilm agent that can be applied as a preservative. In this study, bacteriocin BM173 was successfully expressed in the Escherichia coli expression system and purified by a 2-step method. Furthermore, it exhibited a broad-spectrum antibacterial activity, high thermal stability (121°C, 20 min), and broad pH stability (pH 3-11). Moreover, the minimum inhibitory concentration values of BM173 against E. coli ATCC 25922 and Staphylococcus aureus ATCC 25923 were 14.8 μg/mL and 29.6 μg/mL, respectively. Growth and time-kill curves showed that BM173 exhibited antibacterial and bactericidal activity. The results of scanning electron microscopy and transmission electron microscopy demonstrated that BM173 increased membrane permeability, facilitated pore formation, and even promoted cell lysis. The disruption of cell membrane integrity was further verified by propidium iodide uptake and lactic dehydrogenase release.
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