Our previous work described a clear loss of Escherichia coli (E. coli) membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine) and N,N-dimethylglycine (DMG), but not N,N,N-trimethylglycine (betaine), under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH.

The objective of this study was to assess the effect of supplementation with sodium salt of N,Ndimethylglycine (DMG-Na) on apparent digestibility (AD) in broiler chickens fed low-and high-fat diets. Twenty-eight 1-d-old broiler chickens were fed one of the dietary treatments: a low-fat diet (LF) or a high-fat diet (HF) supplemented with or without 1,000 mg/kg of DMG-Na. Body weight and feed consumption were recorded at 14 and 35 d of age. Average daily growth, daily feed intake, and feed conversion ratio were calculated. The AD of DM, organic matter (OM), CP, total fat (TF), and α-tocopheryl-acetate were assessed by 2 digestibility trials (at 18-21 and 32-35 d, respectively). Serum protein and plasma α-tocopherol concentrations were assessed at 35 d of age. Final BW, feed intake, carcass, breast, and spleen weight were higher in groups fed LF than HF diets (P = 0.048, P = 0.002, P = 0.039, P < 0.001, P = 0.007, respectively). Liver weight was increased in DMG-Na-unsupplemented groups (P = 0.011) for both fat levels. During the first digestibility trial (18-21 d), the AD of DM (P = 0.023), OM (P = 0.033), CP (P = 0.030), and α-tocopheryl-acetate (P = 0.036) was higher in the DMG-Na-supplemented group than control. Digestibility of total fat was increased by DMG-Na supplementation in the LF groups (P = 0.038). A trend for improvement of digestibility was observed during the second digestibility trial (32-35 d) for DM (P = 0.089), OM (P = 0.051), and CP (P = 0.063) in DMG-Na groups. Total serum proteins (and relative fractions) were positively influenced by DMGNa supplementation both in LF and HF diets (P = 0.029). Plasma α-tocopherol concentration was higher in groups fed LF than HF diets (P < 0.001).

The current study first investigates the emulsifying potential of glycine and its N-methylated derivatives N-methylglycine (sarcosine), N,N-dimethylglycine (DMG) and N,N,N-trimethylglycine (betaine) under varying pH conditions. Subsequently, the effect of these test compounds on the membrane integrity of enterotoxigenic Escherichia coli (ETEC) was evaluated. Oil in water emulsions containing each compound show that DMG is a more potent enhancer of emulsification than glycine, sarcosine and betaine under the conditions tested. Flow cytometry was used to investigate whether the emulsifying potential is associated with an effect on ETEC membrane integrity. The bacteria were exposed to each of the test compounds under varying pH conditions and membrane integrity was assessed using the LIVE/DEAD BacLight kit. Results show a membrane deteriorating effect caused by glycine, sarcosine and DMG, but not by betaine. This effect is pH- and time-dependent and has an apparent threshold at pH 9.0. Conventional plate counts confirmed concomitant changes in culturability of the membrane comprised bacteria.

In this study, a total of 200 male broiler chickens (Ross 308) were assigned to four dietary treatments (5 pens/treatment and 10 birds/pen) for two feeding phases: starter (0–11 days of age) and grower-finisher (11–33 days of age). A basal diet containing soy oil (SO) as added fat was used as control group (C), tested against three experimental diets where the SO was partially substituted by BSF larvae fat (BSF) or one of two types of modified BSF larvae fat (MBSF1 and MBSF2, respectively). The two modified BSF larvae fats had a high and low ratio of monobutyrin to monoglycerides of medium chain fatty acid, respectively. Diet did not influence the growth or slaughter performance, pH, color, or the chemical composition of breast and thigh muscles, gut morphometric indices, or histopathological alterations in all the organs. As far as fecal microbiota are concerned, MBSF1 and MBSF2 diets reduced the presence of Clostridium and Corynebacterium, which can frequently cause infection in poultry. In conclusion, modified BSF larva fat may positively modulate the fecal microbiota of broiler chickens without influencing the growth performance and intestinal morphology or showing any adverse histopathological alternations.

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This study was executed to investigate the effect of dietary β-alanine (BA) on amino acid (AA) metabolism and voluntary feed intake in carp (Cyprinus carpio) at mildly elevated temperature to exert AA catabolism. Twenty-four fish in 12 aquaria were randomly assigned to either a control diet or the same diet with 500 mg BA/kg. A 14-day period at an ideal temperature (23 °C) was followed by 15 days at chronic mildly elevated temperature (27 °C). After the 15 days, all fish were euthanised for muscle analysis on histidine-containing dipeptides (HCD), whole blood on free AA and carnitine esters. The carnosine and anserine analysis indicated that all analyses were below the detection limit of 5 µmol/L, confirming that carp belongs to a species that does not store HCD. The increases in free AA concentrations due to BA supplementation failed to reach the level of significance. The effects of dietary BA on selected whole blood carnitine esters and their ratios were also not significant. The supplementation of BA tended to increase body weight gain (P = 0.081) and feed intake (P = 0.092). The lack of differences in the selected nutrient metabolites in combination with tendencies of improved growth performance warrants further investigation to unravel the mechanism of BA affecting feed intake. This first trial on the effect of BA supplementation on AA catabolism showed that its metabolic effect in carp at chronic mildly elevated temperature was very limited. Further studies need to evaluate which conditions are able to exert an effect of BA on AA metabolism.