Aroma active compounds in milk from goat fed basil (Ocimumbasilicum)

The study deals with the addition of basil (Ocimum basilicum) green tops into goat feeding ratio and with the effect of this addition on aroma active compounds in milk. The experiment comprised 8 goats crossbreed of White Goat and Brown Mountain Goat. These goats, which were on the second lactation, were divided into two groups - control (n = 4) and experimental (n = 4). The addition of 0.1 kg of basil green tops into goat feeding ration had no statistically significant effect on differences in total solid, fat or total nitrogen substances in milk during basil green tops feeding. However, the content of aroma active compounds (1-octanol, 1-undecanol, 2-nonanone), which were obtained by simultaneous distillation-extraction (SDE) and determined with usage GC-MS, reached statistically significant differences in days of basil green tops feeding. The amount of 1-octanol, 1-undecanol and 2-nonanone rose from 18.24 ± 0.019,4.48 ± 0.019 and 5.37 ± 0.032 μl/l00ml of milk in days without basil green tops in feeding ration to 24.15 + 10.398,9.60 + 0.118 and 23.29 ± 0.408 ul/lOOml of milk in the first day of basil green tops feeding when the increase was mostly the highest. In the next days of experiment the amount of 1-octanol, 1-undecanol and 2-nonanone was falling despite of readily goats' acceptation of basil green tops

Métabolisme des terpénoïdes chez les caprins

The main objective of this thesis is to study in vitro and in vivo the metabolism of some terpenes characteristic of winter-spring ration ingested by dairy goats in the region of Basilicata (Italy). We sought to quantify as the first step, the metabolism of terpenes in the fermentation broth in vitro and to characterize the major variation factors of their metabolism. We also conducted a test in vivo on 4 individual terpenes (-pinene, -pinene, p-cymène and linalool, the main terpenes ingested by goats in Basilicata), in order to validate the results obtained in vitro, to characterize their changes in the rumen, and their duodenal and blood flow level. In parallel we have studied the impact of these terpenes on ruminal metabolism of cell walls and protein, as well as their effects on the performance of dairy goats. The experimental part is composed of three sets of studies. The methodological studies, in vitro and in vivo studies of metabolism of terpenes. The methodological optimization studies of an extraction and analysis method (SPME "Solid Phase Microextraction") have been conducted in order to obtain the optimum conditions for the extraction of terpenes in the fermentation broth and blood matrix. The first in vitro essay led to a discrimination of tested terpene in terms of their disappearance in the rumen juice, which has been associated to chemical type of considered terpene. The hydrocarbons terpenes showed indeed a much greater disappearance compared to oxygenated ones. This difference is mainly due to differences in their physico-chemical properties, which separates in part their metabolic pathway. The disappearance of terpenes appears to have two main causes, biological intervention of microbial origin and interaction with the matrix. A lower toxic and higher hydrophobic property seems to reinforce the microbial metabolism and the intervention of the matrix on hydrocarbon terpenes compared to those oxygenated ones. In the second test in vitro, the effect of three variables, the type of inoculum (rich in cell walls vs. rich in starch), the type of bacteria (gramme positive vs gramme negative- selected by the contribution of antibiotic) and the redox potential were studied on the metabolism of terpenes in the fermentation broth. The results showed that the inoculum high in cell walls, theoretically supporting cellulolytic bacteria increases the apparent degradation of terpenes. The absence of an effect from the addition of the antibiotic eliminating gramme positive bacteria showed that the gramme negative bacteria are principal responsible, which contribute to the metabolism of terpenes. Finally, the increase in the redox potential increased the disappearance of some of tested terpenes, especially in the inoculum rich in starch. The results from in vitro studies on the metabolism of terpenes, were confirmed by those observed in vivo. However a higher disappearance observed in vivo suggested the involvement of other varying factors that can be attributed to the absorption in the rumen and a loss from rumination and belching. The impact of terpenes on rumen metabolism and performance of dairy goats was studied by incorporating a mixture of four terpenes (mentioned above) at two doses (0.05 and 0.5 ml / kg DMI). Ingestion of terpenes did not produce any effect on the ruminal metabolism of cell walls, and protein. The productivity performance goats were not affected by terpenes, but a change in the milk protein content (a drop in protein content at low dose) observed, can be considered as a secondary effect (effect of dilution) linked to a numerically increase in the milk production at this dose level.L'objectif principal de cette thèse est d'étudier in vitro et in vivo, le métabolisme de certains terpènes caractéristiques de régimes d'hiver-printemps ingérés par les chèvres laitières dans la région du Basilicate (Italie). Nous avons cherché à quantifier dans une première étape le métabolisme des terpènes en milieu fermentaire in vitro, et de caractériser les facteurs majeurs de variation de leur métabolisme. Nous avons également effectué un essai in vivo sur 4 terpènes particuliers (-pinène, -pinène, p-cymène, et linalool, les terpènes principaux ingérés par les chèvres en Basilicate) dans le but de valider les résultats obtenus in vitro, de caractériser leur devenir dans le rumen, et leur flux au niveau duodénal et sanguin. En parallèle nous avons étudié l'impact de ces terpènes sur le métabolisme ruminal des parois et des protéines, ainsi que leurs effets sur les performances des chèvres laitières. La partie expérimentale est composée de trois séries d'études. Les études méthodologiques, les études de métabolisme des terpènes in vitro et celles in vivo. Les études méthodologiques de mise au point d'une méthode d'extraction et d'analyse (SPME «Solid Phase Microextraction») ont été réalisées afin d'obtenir les conditions optimales pour l'extraction des terpènes en milieu fermentaire et dans la matrice sanguine. Le premier essai in vitro a conduit à une discrimination des terpènes testés en termes de leur disparition dans le jus de rumen, qui a été associée au type chimique du terpène considéré. Les terpènes hydrocarbones ont montré en effet une disparition beaucoup plus marquée par rapport aux terpènes oxygénés. Cette différence tient principalement à leurs différences de propriétés physico-chimiques, qui sépare en partie leur voie métabolique. La disparition de terpènes semble avoir deux causes principales, une intervention biologique d'origine microbienne et une interaction avec la matrice. Une propriété moins toxique et plus hydrophobe semble renforcer le métabolisme (dégradation) microbien et l'intervention de la matrice sur les terpènes hydrate de carbones par rapport à ceux oxygénés. Dans le deuxième essai in vitro, l'effet de trois facteurs de variation, le type d'inoculum (riche en fibre vs. riche en amidon), le type des bactéries (Gram + vs Gram -, sélectionnées par l'apport d'antibiotique) et le potentiel redox ont été étudiés sur le métabolisme des terpènes en milieu fermentaire. Les résultats obtenus ont montré que l'inoculum riche en fibre, théoriquement favorisant les bactéries cellulolytiques accroît la dégradation apparente de terpènes. L'absence d'un effet de l'ajout de l'antibiotique éliminant des bactéries Gram + a montré que ce sont les bactéries Gram - qui contribuent au métabolisme des terpènes. Enfin, l'augmentation du potentiel redox a augmenté la disparition de certains des terpènes testés, notamment dans l'inoculum riche en amidon. Les résultats obtenus dans les études in vitro concernant le métabolisme des terpènes, ont été confirmé par ceux observés in vivo. Toutefois une disparition supérieure observée in vivo a suggéré l'intervention d'autres facteurs de variation qui peuvent être attribué à l'absorption dans le rumen et une perte causée par rumination et éructation. L'impact de terpènes sur le métabolisme du rumen et la performance des chèvres laitières a été étudié par l'incorporation d'un mélange de quatre terpènes (mentionnées ci-dessus) à deux doses (0.05 et 0.5 ml/kg MSI). L'ingestion de terpènes n'a provoqué aucun effet sur le métabolisme ruminal des fibres, et des protéines. Les performances de production des chèvres n'ont pas été affectées par les terpènes; cependant une modification du taux protéique (chute du Taux protéique à dose faible) observée, peut être considérée comme un effet secondaire (effet de dilution) lié à une augmentation numérique de la production du lait à cette dos

In vitro degradation by mixed rumen bacteria of 17 mono- and sesquiterpenes typical of winter and spring diets of goats on Basilitica rangelands (southern Italy)

BACKGROUND: Nine monoterpenes (δ-3-carene, p-cymene, limonene, β-myrcene, (E)- and (Z)-β-ocimene, α-phellandrene, α-terpinene, γ-terpinene), seven oxygenated monoterpenes (1,8-cineole, linalool, (E)- and (Z)-linalool oxide, 4-terpinenol, α-terpineol, α-terpinolene) and one sesquiterpene (β-cedrene) were investigated for their degradability in the rumen microbial ecosystem. These molecules were identified as dominant terpenes in the winter and spring diets of milking goats in Basilicata (southern Italy). RESULTS: All terpenes were tested at 3.33 µL L−1 for 24 h using in vitro incubation with mixed rumen bacteria from dairy goats. Oxygen-containing compounds were those recovered at the highest levels (89% of (E)-linalool oxide, 93% of (Z)-linalool oxide, 91% of 1,8-cineole, 82% of terpineol and 72% of 4-terpinenol), except linalool. The linear alkenes β-myrcene and β-ocimene almost completely disappeared. Results were more variable among cyclic alkenes, with recovery rates ranging from 50% in the case of limonene to less than 1% for α-phellandrene. 17% of the only sesquiterpene of the group, β-cedrene, was recovered. CONCLUSION: Recovery rates differed markedly among terpenes, partly in relation to the presence of oxygen and rings in the molecules. These observations should contribute to a better understanding of the changes in composition between the diet and milk terpenes

Optimisation of headspace solid-phase microextraction for quantitative analysis of monoterpenes in caprine blood

In order to follow the metabolic fate of alpha-pinene, beta-pinene, p-cymene and linalool, four monoterpenes characteristic of the spring diet of dairy goats in the Basilicata region (Southern Italy), we have investigated the use of headspace solid-phase microextraction (HS-SPME) on a 100 mu m polydimethylsiloxane coated fibre as a preparation technique for their analysis by gas chromatography in blood. The effects on terpenes recovery yields of exposure temperature, between 25 and 45 degrees C, exposure time, between 5 and 25 min, and sample ionic strength, between 0 and 250 g/L NaCl, were modelled by second order polynomials parameterised with data from a Hoke experimental design of 13 runs. HS-SPME of alpha- and beta-pinene was mostly favoured by low temperatures and, to a lesser extent, by short extraction times in a linear way. The model fitted to p-cymene data was characterised by a curvilinear effect of extraction time (main effect, P = 0.04; quadratic effect, P=0.10) and quadratic effect of salting (P = 0.04) generating a saddle-shaped surface response. Peak areas of p-cymene were lowered by intermediate NaCl concentrations and short extraction times. For linalool, all the main effects (P < 0.04) and the quadratic effect of extraction time (P = 0.02) generated a hill-shaped response surface. The largest linalool peaks resulted from relatively low extraction times and high temperatures. The best set of operative conditions was an exposure temperature of 35 degrees C. an exposure time of 22 min and a NaCl concentration of 250 g/L. With limonene as an internal standard, the linearity, intra-assay precision and quantification limits were estimated for each terpene determination. The proposed HS-SPME method allowed fast, simple and sensitive determination of terpenes in goat blood samples by conventional equipment. (C) 2012 Elsevier B.V. All rights reserved

Degradation of terpenes and terpenoids from Mediterranean rangelands by mixed rumen bacteria in vitro

This in vitro study aimed at estimating the disappearance rates of 14 terpenes and terpenoids after 24-h incubation with mixed bacteria from caprine rumens. These compounds comprised nine monoterpene hydrocarbons (d-3-carene, p-cymene, b-myrcene, (E)- and (Z)-b-ocimene, a-phellandrene, a-terpinene, g-terpinene and a-terpinolene), four oxygenated monoterpenes ((E)- and (Z)-linalool oxide, 4-terpinenol, a1g terpineol) and one sesquiterpene hydrocarbon (b-cedrene). They were individually exposed to goat rumen microflora for 24 h in 70ml culture tubes at an input level of 0.5 ml/l. Terpenoids were the least degraded, 100% of (E)-linalool oxide, 95% of (Z)-linalool oxide, 91% of 4-terpinenol and 75% of terpineol remained intact after 24-h incubation. In contrast, a-terpinolene concentration in fermentation broth extracts was below quantification limit, thus indicating an extensive, if not complete, degradation by rumen bacteria. Only 2% of the initial amounts of a-phellandrene were recovered. The other monoterpenes and b-cedrene were partly degraded, with losses ranging from 67% for d-3-carene to 90% for (E)-b-ocimene. The corresponding rates of disappearance were between 2.67 and 4.08 mmol/ml inoculum per day

Dégradabilité ruminale de monoterpènes présents dans le régime de printemps de la chèvre laitière dans le Basilicate (italie)

National audienc

In vitro and in sacco determining the nutritive value of button mushroom stipe and its application in growing lambs diet

Feed shortage is one of the major challenges in livestock production systems in the arid and semi-arid regions of the world. Hence, wastes of nutritional value from the agro-alimentary industry could be considered as by-product feedstuffs, reducing diet costs in these regions. The current study aimed at determining in vitro and in sacco nutritive value of button mushroom stipe (BMS) and investigating its potential in growing lambs diet. For this purpose, three experiments (Exp.) were conducted. Ruminal degradability characteristics, and gas production kinetics, and energetic value of BMS compared with alfalfa were determined by the nylon bag and gas production techniques in Exp.1 and Exp.2, respectively. In Exp.3, 24 Mehraban growing lambs were assigned randomly to one of the three dietary treatments: (1) the basal ration without BMS (CTRL); (2) and (3) inclusion of 10 and 20% (DM basis) of BMS in the basal ration (BMS10 and BMS20, respectively), to determine their growth performance and the rations digestibility. The soluble (a) and slowly degradable (b) fractions, and degradation rate of ‘b’ fraction of the BMS crude protein (CP) was 43, 59%, and 0.034, respectively. The metabolisable energy (ME) content of BMS was 20.6% lower than that in alfalfa. The inclusion of BMS in the ration of lambs did not affect their growth performance, but reduced diet cost by 7.2 and 14.5% in BMS10 and BMS20, respectively. The BMS tended to increase (p = .089) the blood total protein in lambs fed BMS10 and tended to decrease (p = .07) alkaline phosphatase (ALP) in BMS-based diets. These results reveal that BMS has a slow ruminal degradability and fermentation that can reduce diet cost by partially replacing conventional feedstuffs in the ruminant diet without adversely affecting their health and performance.Highlights The button mushroom stipe (BMS) by-product has relatively high crude protein and acceptable energy contents with a slow ruminal degradability. Including BMS in the diet of growing lambs reduced diet costs, while improving fairly their growth performance, implying that BMS can be considered as a cost-effective by-product feedstuff in a ruminant diet. The inclusion of BMS in the diet of lambs decreased ruminal ammonia production, which can reduce the environmental impact of ruminant production systems