The absorption and metabolism of phytogenic feed additives in poultry is studied related to the metabolism and deposition of their main compounds in tissues intended for food production. Fifty-six non-sexed Ross 308 broilers were allocated to seven dietary treatments and fed a diet containing graded levels of thyme (Thymus vulgaris L.) essential oil (EO) (0, 0.01, 0.02, 0.03, 0.04, 0.05, 0.1%, w/w). Thymol concentration was measured in plasma, liver, kidney and breast muscle tissue using solid phase micro-extraction followed by gas chromatography/mass spectrometry. We found the highest concentrations of thymol in kidney and plasma, and the lowest in breast muscle and liver. Thymol content in plasma and kidney significantly increased when 0.05 and 0.1%, w/w, EO and in liver and breast muscle only when 0.1%, w/w, EO was added to the diet (p<0.05). Our results indicate intensive metabolism of thymol in liver and its accumulation in kidney tissue. We confirm low deposition of thymol in the muscle tissue. It is necessary to keep in mind the selection of a sufficient concentration of EO in the feed additive for animals without the risk of thymol residues in edible tissues.
Two experiments were conducted on sheep to determine the effect of dietary supplementation with zinc and a medicinal plant mixture on haematological parameters and microbial activity in the rumen and large intestine. In Experiment 1, 24 male lambs were randomly divided into four groups: One group was fed an unsupplemented basal diet (control), and three groups were fed a diet supplemented with 70 mg Zn/kg diet in the form of Zn sulphate (ZnSO ), a Zn-chelate of glycine hydrate (Zn-Gly) or a Zn-proteinate (Zn-Pro), for five months. The ruminal content was collected separately from each lamb, and batch cultures of ruminal fluid were incubated in vitro with mixture of medicinal plants (Mix) with different roughage:concentrate ratios (800:200 and 400:600, w/w). Bioactive compounds in Mix were quantified by UPLC/MS/MS. In Experiment 2, four sheep were fed a diet consisting of meadow hay and barley grain (400:600, w/w), with Zn-Gly (70 mg Zn/kg diet), Mix (10% replacement of meadow hay) or Zn-Gly and Mix (Zn-Gly-Mix) as supplements in a Latin square design. Mix decreased total gas (p < 0.001) and methane (p < 0.01) production in vitro. In Experiment 1, caecal isobutyrate and isovalerate concentrations varied among the dietary treatments (p < 0.01). The isovalerate concentration of the zinc-supplemented groups in the distal colon was higher (p < 0.001) compared with the control. In Experiment 2, the molar proportion of isobutyrate was the highest in the faeces of the sheep fed the diet with Zn-Gly-Mix (p < 0.01). The plasma zinc concentration was higher in the groups fed a diet supplemented with zinc (p < 0.001). The haematological profile and antioxidant status did not differ between the dietary groups (p > 0.05). The diets containing medicinal plants and organic zinc thus helped to modulate the characteristics of fermentation in ruminants.
In the present study, we aimed to develop a method for thymol sulfate and thymol glucuronide determination in plasma, liver and duodenal wall of broiler chickens after feeding with a Thymus vulgaris essential oil at the different concentrations (0.01, 0.05 and 0.1% w/w). UHPLC coupled with accurate-mass QTOF-MS was used for identification and quantification of thymol metabolites. Novel Waters Oasis Prime HLB solid-phase extraction cartridges were applied to sample clean-up with extraction recoveries ranged from 85 to 92%. The presence of thymol glucuronide was confirmed by MS software according to molecular formula, score, mass error and double bond equivalent. In terms of validation, calibration curves of thymol sulfate were constructed in matrix samples with linearity from 3.91 to 250.0 ng/mL and correlation coefficients were within the range of 0.9979-0.9995. Limits of detection were 0.97, 0.29 and 0.63 ng/mL and limits of quantification were 3.23, 0.97 and 2.09 ng/mL for plasma, liver and duodenal wall, respectively. Intra-day and inter-day precision expressed as relative standard deviation were <4.35%. To highlight, thymol metabolites were directly detected for the first time in liver and duodenal wall and this method was shown to be successfully applicable for investigation of thymol metabolism in chickens after thyme essential oil ingestion.
Plant compounds occurring in phytogenic feed additives are involved in different pharmacological activities in the animal organism. Since the digestive tract acts as a first line of defence against foreign compounds, it is necessary to outline its response to dietary supplementation with bioactive plant components. Little information is available on the bioactivity of thymol as the main bioactive compound of Thymus vulgaris L. essential oil (TEO). The main objective of the present study was to provide a detailed view of the concentrations of thymol in plasma and the content of individual intestinal segments (duodenum, jejunum, ileum, caecum and colon) of broiler chickens after 4 weeks of dietary supplementation with different TEO concentrations. 32 one‐day old Ross 308 hybrid broilers were randomly divided into four dietary treatment groups (0.00%, 0.01%, 0.05%, 0.1% w/w of TEO in the diet). Thymol concentrations in the duodenal chyme presented around 7% on average from the thymol amount administered in the feed. A significantly increased thymol amount was observed after 0.1% TEO addition to the diet compared with 0.01% TEO enrichment in the duodenal wall and gut content of jejunum, ileum, caecum and colon (p < 0.05). Thymol levels in the colon were significantly higher than in the ileum and about 1.7 times higher on average than those in the caecum. Significant coefficient of correlation was observed between thymol concentrations in plasma and feed, gut content of all intestinal segments as well as duodenal wall. Our results point to intensive thymol absorption in the initial sections of the digestive tract. In the current study, the role of intestine in biotransformation of thymol was observed, and it would be desirable to investigate whether thymol itself or thymol metabolites are responsible for beneficial effects in intestine.
The abomasal parasitic nematode Haemonchus contortus can influence the abomasal microbiome of the host. On the other hand, no information occurs on the influence of the parasite on the hindgut microbiome of the host. We evaluated the impact of Haemonchus contortus on the fecal microbial community of the experimentally infected lambs treated with a mixture of medicinal herbs to ameliorate the haemonchosis. Twenty-four female lambs were divided into four groups: infected animals (Inf), infected animals supplemented with a blend of medicinal herbs (Inf+Herb), uninfected control animals (Control), and uninfected animals supplemented with medicinal herbs (C+Herb). Inf and Inf+Herb lambs were infected orally with approximately 5000 L3 larvae of a strain of H. contortus susceptible to anthelmintics (MHco1). Herb blend (Herbmix) consisted of dry medicinal plants of Althaea officinalis,
Two 24 h in vitro batch culture experiments were conducted to investigate the effects of fumarate addition (10 mmol L −1 ) on the ruminal fermentation parameters of selected medicinal herbs, and the effects of different doses of fumarate (0, 10, or 30 mmol L −1 ) on ruminal metabolism of organic acids when a high-concentrate diet (meadow hay and barley grain, 400/600, w/w) was supplemented with a mix of medicinal herbs (Artemisia absinthium L., Melissa officinalis L., Malva sylvestris L., Matricaria chamomilla L., Plantago lanceolata L., Foeniculum vulgare Mill., and Althaea officinalis L.). Depending on the concentration, fumarate treatment decreased methane production (by 10-11%) and increased propionate proportions (by 5-13%) with high-concentrate diets. The organic acid (fumaric, succinic, malic, and lactic acid) concentrations in the batch culture were measured at intervals of 0, 4, 6, 12, and 24 h. The time and organic acid concentrations with 10 mmol L −1 fumarate were well correlated (R 2 = 0.846). The amount of succinate was accumulated and metabolized more slowly than that of fumarate (>24 h). The addition of fumarate and a herb mix could positively influence in vitro ruminal fermentation parameters of high-concentrate diets by increasing the levels of propionate and succinate as well as the pH, and by decreasing of methane emissions.
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