Gastrointestinal metabolism of phytoestrogens in lactating dairy cows fed silages with different botanical composition.

Dietary phytoestrogens are metabolized or converted in the gastrointestinal tract of ruminants, only limited knowledge exists on the extent and location of this conversion in vivo. The objective of this study was to quantify the gastro-intestinal metabolism of phytoestrogens in lactating dairy cows fed silages with different botanical composition. Four lactating rumen cannulated Norwegian Red cattle were assigned to a 4 × 4 Latin square with 1 cow per treatment period of 3 wk. The 4 treatment silages were prepared from grasslands with different botanical compositions: organically managed short-term timothy (Phleum pratense L.) and red clover (Trifolium pratense L.) ley (2 yr old: ORG-SG); organically managed long-term grassland with a high proportion of unsown species (6 yr old; ORG-LG); conventionally managed perennial ryegrass (Lolium perenne L.) ley (CON-PR); and conventionally managed timothy ley (CON-TI). The herbages were cut, wilted, and preserved with additive in round bales, fed as a mix of the first and third cut at 90% of ad libitum intake, and contributed to 70% of the total dry matter intake. Milk, feed, omasal digesta, urine, and feces were collected at the end of each period and analyzed for the concentrations of phytoestrogens by using a liquid chromatography-tandem mass spectrometry technique. Concentration of total isoflavones was highest in ORG-SG and lowest in CON-TI silage, whereas the content of total lignans was highest in the grass silages. The isoflavones were extensively metabolized in the rumen on all diets, and the recovery of formononetin and daidzein in omasum, mainly as equol, averaged 0.11 mg/mg. The apparent intestinal metabolism was less severe as, on average, 0.29 mg/mg of the omasal flow was recovered in feces. The plant lignans were also strongly degraded in the rumen. However, the flow of lignans to omasum and excretion in feces were, on average, 7.2- and 5.2-fold higher, respectively, than the intake of the plant lignans matairesinol and secoisolariciresinol, known as precursors of mammalian lignans. Thus, excretion to milk could not be directly related to intake, implying that plant lignans other than matairesinol and secoisolariciresinol in forage are precursors for enterolactone production in the rumen and for its content in milk. Equol followed mainly the flow of large particles out of the rumen, whereas the mammalian lignans were distributed between phases proportional to dry matter flow. The main metabolism of phytoestrogens occurred in the rumen and the main route of excretion was through feces and urine, with only a small part being excreted in milk. The concentration of phytoestrogens in milk can be manipulated through intake but the intermediate transfer capacity to milk appears to be limited by saturation.

Effect of silage botanical composition on ruminal biohydrogenation and transfer of fatty acids to milk in dairy cows.

Ruminal biohydrogenation and transfer of fatty acids (FA) to milk were determined for 4 silages with different botanical compositions using 4 multiparous Norwegian Red dairy cows [(mean ± SD) 118 ± 40.9 d in milk, 22.5 ± 2.72 kg of milk/d, 631 ± 3.3 kg of body weight, 3.3 ± 0.40 points on body condition score at the start of the experiment] fitted with rumen cannulas. Treatments consisted of 4 experimental silages: a mix of the first and third cut of organically managed short-term grassland with timothy (Phleum pratense L.) and red clover (Trifolium pratense L.; 2 yr old; ORG-SG); organically managed long-term grassland with a high proportion of unsown species (6 yr old; ORG-LG); conventionally managed ley with perennial ryegrass (Lolium perenne L.; CON-PR); and conventionally managed ley with timothy (CON-TI). The herbages were cut, wilted, and preserved with additive in round bales and fed at 0.90 of ad libitum intake. A barley (Hordeum vulgare L.) concentrate constituted 300 g/kg of dry matter of the total feed offered. A Latin square design (4 × 4) with 3-wk periods and the last week in each period used for sampling was implemented. Omasal flows of FA were measured using Yb acetate, Cr-EDTA, and the indigestible neutral detergent fiber fraction as indigestible markers. The composition of FA was analyzed in feed, omasal digesta, and milk. Compared with ORG-LG, ORG-SG had a higher herbage proportion of red clover (0.36 vs. 0.01) and lower proportions of timothy (0.42 vs. 0.18), smooth meadowgrass (Poa pratensis L.), meadow fescue (Festuca pratensis Huds.), white clover (Trifolium repens L.), dandelion (Taraxacum spp.), and creeping buttercup (Ranunculus repens L.). The silages were well preserved. The concentration of neutral detergent fiber was higher and the concentration of Kjeldahl-N was lower for CON-TI than for the other silages. Silage type had no effect on dry matter intake, but milk yield was lower for CON-TI than for the other silages. Apparent biohydrogenation of C18:3n-3 was lower for ORG-SG (932 g/kg) than for ORG-LG (956 g/kg), CON-PR (959 g/kg), and CON-TI (958 g/kg). Compared with the grass-based silages, ORG-SG and ORG-LG resulted in higher omasal flows of C18:1 trans FA and higher milk fat proportions of C18:1 trans FA and C18:2 cis-9,trans-11. Apparent recovery of C18:3n-3 in milk was higher for ORG-SG (61 g/kg) than for ORG-LG (33 g/kg), CON-PR (34 g/kg), and CON-TI (38 g/kg), and milk fat proportion of C18:3n-3 was higher for ORG-SG than for CON-TI. Milk fat proportions of C16:0 were lower for ORG-SG and ORG-LG compared with those for CON-PR and CON-TI. It was concluded that high proportions of red clover and other dicotyledons in the silages affected ruminal biohydrogenation and increased milk fat proportions of beneficial FA.

The effect of N-fertilisation rate or inclusion of red clover to timothy leys on fatty acid composition in milk of dairy cows fed a commercial silage: concentrate ratio.

The aim of this experiment was to, under typical Swedish production conditions, evaluate the effects of grass silages subjected to different N-fertilisation regimes fed to dairy cows on the fatty acid (FA) composition of their milk, and to compare the grass silages in this respect to red clover-dominated silage. Grass silages made from first year Phleum pratense L. leys subjected to three N-fertilisation regimes (30, 90 and 120 kg N/ha, designated G-30, G-90 and G-120, respectively) and a mixed red clover-grass silage (Trifolium pratense L. and P. pratense L.; 60/40 on dry matter (DM) basis, designated RC-G) were produced. The experiment was conducted as a change-over design, including 24 primiparous and multiparous dairy cows of the Swedish Red breed, each of which was allocated to three of the four diets. The cows were offered 11 kg DM of silage and 7 kg concentrates. The silages had similar DM and energy concentrations. The CP concentration increased with increase in N-fertilisation level. There was a linear increase in DM intake of the different silages with increased N fertilisation. There were also differences in concentrations of both individual and total FAs amongst silages. The daily milk production (kg/day) did not significantly differ between treatments, but G-30 silage resulted in higher concentrations of 18:2n-6 in the milk compared with the other two grass silages. The highest concentrations of 18:3n-3 and cis-9, trans-11 18:2 were found in milk from cows offered the RC-G silage. The G-30 diet resulted in higher concentration of 18:2n-6 and the same concentration of 18:3n-3 in the milk as the other grass silages, despite lower intake levels of these FAs. The apparent recoveries of 18:3n-3 from feed to milk were 5.74%, 4.27%, 4.10% and 5.31% for G-30, G-90, G-120 and RC-G, respectively. A higher recovery when red clover is included in the diet confirms previous reports. The higher apparent recovery of 18:3n-3 on the G-30 treatment may be related to the lower silage DM intake, which led to a higher relative proportion of ingested FAs originating from concentrates compared with the G-90 and G-120 diets. With the rates and types of concentrates used in this study, the achieved differences in FA composition among the silages were not enough to influence the concentrations of unsaturated FAs in milk.

Effects of feeding dairy cows different legume-grass silages on milk phytoestrogen concentration.

Phytoestrogens are hormone-like substances in plants that can substantially influence human health (positively or negatively), and when fed to dairy cows are partly transferred to their milk. The aim of this study was to investigate the effects of varying the botanical composition and regrowth interval of legume-grass silage on phytoestrogen intake and milk phytoestrogen concentrations. In one experiment, 15 Swedish Red dairy cows were fed 2- or 3-cut red clover-grass silage, or 2-cut birdsfoot trefoil-grass silage. In a second experiment, 16 Norwegian Red dairy cows were fed short-term ley silage with red clover or long-term ley silage with white clover, and the effects of supplementation with α-tocopherol were also tested. High concentrations of formononetin and biochanin A were found in all silage mixtures with red clover. The milk concentration of equol was highest for cows on the 2-cut red clover-grass silage diet (1,494 µg/kg of milk). Because of the metabolism of biochanin A, genistein, and prunetin, their concentrations in milk and the apparent recovery were low. Coumestrol was detected in only short-term and long-term ley silage mixtures, and its milk concentration was low. Concentrations of secoisolariciresinol and matairesinol were higher in 2-cut birdsfoot trefoil-grass and long-term ley silage mixtures, those with legume species other than red clover, and the highest grass proportions. The 2-cut birdsfoot trefoil-grass silage diet also resulted in higher enterolactone concentration than the other diets (226 µg/kg of milk). Lengthening the regrowth interval increased the intake of secoisolariciresinol and decreased the recovery of lignans. Feeding long-term ley silage resulted in higher milk lignan concentrations but lower milk isoflavone concentrations than feeding short-term ley silage. The apparent recovery of all phytoestrogens except prunetin was highest on the 2-cut birdsfoot trefoil-grass silage diet. No effect of α-tocopherol supplementation was observed on milk concentrations of any of the measured phytoestrogens. Variations were observed in milk concentrations of phytoestrogens, especially of equol, among cows, which could not be explained by variations in diet composition or phytoestrogen intake. The results show that milk phytoestrogen concentration is strongly influenced by silage botanical composition, but questions regarding phytoestrogen metabolism remain to be answered.

First reported foodborne outbreak associated with microsporidia, Sweden, October 2009.

Microsporidia are spore-forming intracellular parasites that infrequently cause disease in immunocompetent persons. This study describes the first report of a foodborne microsporidiosis outbreak which affected persons visiting a hotel in Sweden. Enterocytozoon bieneusi was identified in stool samples from 7/11 case-patients, all six sequenced samples were genotype C. To confirm that this was not a chance finding, 19 stool samples submitted by healthy persons from a comparable group who did not visit the hotel on that day were tested; all were negative for microsporidia. A retrospective cohort study identified 135 case-patients (attack rate 30%). The median incubation period was 9 days. Consumption of cheese sandwiches [relative risk (RR) 4·1, 95% confidence interval (CI) 1·4-12·2] and salad (RR 2·1, 95% CI 1·1-4) were associated with illness. Both items contained pre-washed, ready-to-eat cucumber slices. Microsporidia may be an under-reported cause of gastrointestinal outbreaks; we recommend that microsporidia be explored as potential causative agents in food- and waterborne outbreaks, especially when no other organisms are identified.

Methane emissions from a constructed wetland treating wastewater--seasonal and spatial distribution and dependence on edaphic factors.

Constructed wetlands for wastewater treatment have many advantages. They can be used for several purposes, for example, to reduce levels of organic matter and nutrients, and to retain toxic metals. However, most wetlands are inherently net sources of gaseous compounds like methane and nitrous oxide, which are of environmental concern due to their rapid current accumulation in the atmosphere and their potent global warming capacity. In order to determine the flux of methane from a constructed wetland a study was conducted over two growth seasons on a pilot scale wetland constructed to reduce nutrient levels in secondary treated wastewater. The emissions for the spring to autumn period averaged 141 mg CH(4)m(-2)d(-1) (S.D.=187), ranging from consumption of 375 mg CH(4)m(-2)d(-1) to emissions of 1739 mg CH(4)m(-2)d(-1). The spatial and temporal variations were large, but could be accounted for by measured environmental factors. Among these factors, sediment and water temperatures were significant in all cases and independent of the scale of analysis (r(2) up to 0.88).