Identification of the botanical origin of raw spirits produced from rye, potato, and corn based on volatile compounds analysis using a SPME-MS method

Determination of the botanical origin of raw spirit used for alcoholic beverage production is of great importance for rectifying units, control laboratories, and proper product labeling. Raw spirit samples (138) produced from rye, corn, and potato were analyzed using a solid phase microextraction-mass spectrometry (SPME-MS) method, which involved volatiles preconcentration by SPME with subsequent volatile fraction characterization by MS without particular compounds separation by GC. Obtained data were treated using principal component analysis and linear discriminant analysis (LDA) to test the possibility of sample classification. SPME sampling conditions allowed rapid extraction in 2 min at 50 °C using a carboxen/divinylbenzene/polydimethylsiloxane fiber, followed by rapid MS analysis. Use of LDA made possible the classification of raw spirits based on the material they were produced from. The classification ability of the developed SPME-MS method was 100%, whereas its prediction ability was 96%.     

Caudal vena cava thrombosis-like syndrome in a horse

A 9-year-old Quarter horse was presented for chronic refractory pneumonia. On necropsy, an hepatic abscess, caudal vena cava thrombosis, pulmonary thromboembolism, and embolic pneumonia were identified. Similar lesions have been reported in cattle as caudal vena cava thrombosis syndrome, however this syndrome has not previously been reported in horses.     

Assessment of disease severity and outcome of dietary, antibiotic, and immunosuppressive interventions by use of the canine IBD activity index in 21 dogs with chronic inflammatory bowel disease

Recently, the canine IBD activity index (CIBDAI) was developed for evaluation of the severity of illness, therapeutic strategies, and efficacy of therapy.The aim of the present study was to assess the severity of illness and the therapeutic strategy in dogs with IBD by the use of CIBDAI, serum albumin concentration, and histologic score (HPEG). Furthermore the use of CIBDAI and the efficacy of therapy in a prospective study during a 3 month treatment period were evaluated. Twentyone dogs with inflammatory bowel disease (lymphocytic-plasmacytic enteritis and enterocolitis) were examined in this study. In 11 dogs with IBD the severity of illness was assessed as low, according to CIBDAI and HPEG (CIBDAI score 4 or between 5 and 10 with HPEG score between 1 and 1.5). Six dogs were treated with hypoallergenic diet (Group D), five dogs were treated with hypoallergenic diet and metronidazole (15.6-22,3 mg/kg/day) (Group M). In 10 dogs with IBD the severity of illness was assessed as high (CIBDAI <10, or CIBDAI between 5 and 10 with HPEG score between 2 and 3 or hypoalbuminemia (< or = 2.5 g/dl)). This group (Group I) was treated with immunosuppressive therapy.Treatment consisted of prednisolone (n=10; 0.9-2 mg/kg/day), azathioprine (n=5; 0.9-2.3 mg/kg/day), sulfasalazine (n=4; 18.2-25 mg/kg/day) and hypoallergenic diet (n=10). Efficacy of therapy was evaluated prospectively 3 times in a 12 weeks treatment period. Remission (CIBDAI score < 4) indicated good therapeutic response, chronic or recurrent disease (CIBDAI score persistent or recurrent > or =4) indicated poor therapeutic response. Age, CIBDAI score and HPEG score were significantly different in IBD dogs with low severity of illness (age: median 60 months; CIBDAI score: median 5; HPEG score: median (1) and IBD dogs with high severity of illness (age: median 90 months; CIBDAI score: median 9.5; HPEG score: median 2.25) (p = 0.0101 and p = 0.0099, respectively). The presence of hypoalbuminemia was not significantly different between these two groups (p = 0.3108). There was no significant correlation between CIBDAI score and serum albumin concentration (r = 0.0394; p = 0.0802) or between CIBDAI score and HPEG score (r = 0.2587; p = 0.2574). In the treatment groups, HPEG score was only significantly different between D-group and group I (p < 0.01). The CIBDAI score decreased significantly in group I after 4 weeks of treatment (median 4th week: 3; p < 0.05), and in the D-group after 8 weeks of treatment (median 8" week 1; p < 0.05). No significant decrease of CIBDAI score was seen in the M-group (median 12th week: 1.75; p > 0.05). All dogs in group D, four of five dogs in group M, and six from ten dogs in group I went into remission. Poor therapeutic response (1 dog in group M and 5 dogs in group I; one dog died) was seen in 6 dogs, where as 15 dogs showed good therapeutic response. There was no significant association between efficacy of therapy and age (p = 0.8455), CIBDAI score (p = 0.3293), or serum albumin concentraton (p = 0.8455). Poor therapeutic response was weekly associated with HPEG score > or =2 (p = 0.0635). Using CIBDAI in dogs with IBD as a single parameter to assess the severity of illness and the therapeutic response, misinterpretations are possible.The assessment of the severity of illness by the combination of CIBAI, HPEG, and serum albumin concentration is leading to adaequate therapeutic results. Dogs with low grade IBD benefit from hypoallergenic diet, whereas dogs with high grade IBD benefit from immunosuppressive therapy. The effect of antibiotic treatment is questionable.     

Application of stable carbon isotope analysis to the detection of testosterone administration to cattle

The use of anabolic substances is prohibited in food-producing animals throughout the European Union. No method is available to reliably detect the misuse of natural hormones in cattle. A method was developed to detect the abuse of testosterone in cattle fattening. Synthesized testosterone is rather depleted in the (13)C/(12)C ratio. Hence, the method is based on gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) analysis of urine. To select testosterone metabolites and endogenous reference compounds (ERC), the concentration of urinary steroids of cattle was investigated. Dehydroepiandrosterone and androst-5ene-3beta,17alpha-diol were chosen as ERCs to show endogenous (13)C/(12)C ratios. Etiocholanolone and 5alpha-androstane-3beta,17alpha-diol were chosen as the most important testosterone metabolites. Other metabolites known from literature like epitestosterone were less promising. In principle, GC/C/IRMS is a nonspecific method because finally carbon dioxide is analyzed. Therefore, a dedicated cleanup procedure for the selected steroids was developed. By means of proposed confidence intervals in the isotopic composition of ERCs and metabolites, the administration of testosterone to cattle could be detected reliably. Differences of up to 11 per thousand on the delta-scale between ERC and testosterone metabolites were found after testosterone administration, whereas endogenous differences did not exceed 2 per thousand.     

Sulfur and nitrogen supply influence growth,product appearance,and glucosinolate concentration of broccoli

The effects of insufficient and optimal sulfur (S) and nitrogen (N) supply on plant growth and glucosinolate formation were studied under controlled experimental conditions in broccoli “Monaco”. Here, we report on the interaction between S and N supply, plant growth, and quality parameters and discuss the relevance of this interaction in relation to crop‐management strategies. Broccoli plants supplied with insufficient amounts of S or N showed typical deficiency symptoms and yield decreases. In contrast, total glucosinolate concentrations were high at insufficient N supply, independent of the S level, and low at insufficient S supply in combination with an optimal N supply. This was mainly due to the presence of the alkyl glucosinolates glucoraphanin and glucoiberin. Furthermore, with S concentrations above 6 g (kg DM)^–1 and an N : S ratio lower than 10:1, the glucosinolate concentrations were on average around 0.33 g (kg fresh matter)^–1 and differed significantly from those plants characterized by an S concentration below 6 g (kg DM)^–1 and an N : S ratio above 10:1. In addition, N : S ratios between 7:1 and 10:1 promoted plant yield and enhanced overall appearance. Therefore, to produce broccoli (and potentially other Brassicaceae) with higher crop yields and enhanced product quality in the field, it is vital to establish the optimal S and N nutritional status of the plant and to integrate this information into crop‐management strategy programs.     

Structural investigations of flavonol glycosides from sea buckthorn (Hippophaë rhamnoides) pomace by NMR spectroscopy and HPLC-ESI-MS(n)

Four flavonol glycosides were isolated from an extract of sea buckthorn pomace (Hippopha? rhamnoides) by Sephadex LH-20 gel chromatography and semipreparative HPLC. Their structures were elucidated by hydrolysis studies, ESI-MS(n), UV, and (1)H and (13)C NMR spectroscopy. The occurrence of the major flavonol glycoside kaempferol 3-O-beta-sophoroside-7-O-alpha-rhamnoside in sea buckthorn is described here for the first time. A further 21 flavonol glycosides of Sephadex LH-20 fractions of sea buckthorn pomace were characterized by HPLC-DAD-ESI-MS. The characteristic MS-MS and MS(3) fragmentation pattern of flavonol glycosides previously identified in sea buckthorn juice and of flavonol glycosides identified by NMR spectroscopy gave valuable indications for their identification. The results demonstrate that loss of the sugar moiety from C-7 of the aglycon is more favored than fission of the glycosidic linkage at the C-3 position. Thus, most of the compounds identified were 7-rhamnosides of isorhamnetin, kaempferol, and quercetin, which exhibit different substitution patterns at the C-3 position, mainly glucosides, rutinosides, and sophorosides. In addition, numerous flavonol glycosides were detected lacking a sugar moiety at C-7. Finally, eight flavonol derivatives were identified that are acylated by hydroxybenzoic or hydoxycinnamic acids.     

Organic carbon accumulation in a 2000-year chronosequence of paddy soil evolution

Considerable amounts of soil organic matter (SOM) are stabilized in paddy soils, and thus a large proportion of the terrestrial carbon is conserved in wetland rice soils. Nonetheless, the mechanisms for stabilization of organic carbon (OC) in paddy soils are largely unknown. Based on a chronosequence derived from marine sediments, the objectives of this study are to investigate the accumulation of OC and the concurrent loss of inorganic carbon (IC) and to identify the role of the soil fractions for the stabilization of OC with increasing duration of paddy soil management. A chronosequence of six age groups of paddy soil formation was chosen in the Zhejiang Province (PR China), ranging from 50 to 2000 years (yrs) of paddy management. Soil samples obtained from horizontal sampling of three soil profiles within each age group were analyzed for bulk density (BD), OC as well as IC concentrations, OC stocks of bulk soil and the OC contributions to the bulk soil of the particle size fractions. Paddy soils are characterized by relatively low bulk densities in the puddled topsoil horizons (1.0 and 1.2 g cm^− 3) and high values in the plow pan (1.6 g cm^− 3). Our results demonstrate a substantial loss of carbonates during soil formation, as the upper 20 cm were free of carbonates in 100-year-old paddy soils, but carbonate removal from the entire soil profile required almost 700 yrs of rice cultivation. We observed an increase of topsoil OC stocks from 2.5 to 4.4 kg m^− 2 during 50 to 2000 yrs of paddy management. The OC accumulation in the bulk soil was dominated by the silt- and clay-sized fractions. The silt fraction showed a high accretion of OC and seems to be an important long-term OC sink during soil evolution. Fine clay in the puddled topsoil horizon was already saturated and the highest storage capacity for OC was calculated for coarse clay. With longer paddy management, the fractions < 20 μm showed an increasing actual OC saturation level, but did not reach the calculated potential storage capacity.     

Influence of sulfur supply,ammonium nitrate ratio,and arbuscular mycorrhizal colonization on growth and composition of Chinese chive

Tissue concentrations of phytochemicals formed by Allium species may be influenced by plant nutrient supply or rhizosphere microorganism activity. To test these relations, three plant nutritional factors were varied in two experiments with Chinese chive [Allium tuberosum]: (a) increasing sulfur concentrations in the substrate, (b) two ratios of ammonium to nitrate in supply, and (c) inoculation with arbuscular mycorrhizal (AM) fungi. Shoot dry weight, nutrient composition (nitrate, N, P, S), and organosulfur compounds (measured indirectly as pyruvic acid) were determined.     

Profiles of maternal origin factors during transition from embryonic diapause to implantation in roe deer

The aim was to evaluate in female roe deer: (a) PAG mRNA relative abundance in endometrial uterine tissue for determination of the duration of embryonic diapause, (b) mRNA relative abundance of progesterone, estradiol, and prolactin (P4, E2, and PRL) receptors (PGR, ESR, and PRLR) during diapause and after implantation in the endometrium; (c) concentration of P4, E2, and PRL in the blood, and (d) a noninvasive method of hormone detection by measurement of P4 and E2 concentrations in feces. A total of fifteen individuals were obtained post mortem during hunting seasons and divided into three experimental groups (November, December, January). The results did not reveal mRNA relative abundance for PAGs in the endometrium or detectable PAG concentrations in the serum of all examined females. Concentration of PRL and mRNA relative abundance for PRLR long isoform in the endometrium was the highest in January (p < .01). mRNA relative abundance for PGR, P4 concentration in the endometrium, serum, and feces was the highest in January (p < .01). Endometrial origin PRL and P4 may be responsible for the termination of this process and pregnancy development after implantation.     

Genetic identity of three mint accessions stored by different conservation procedures: field collection, in vitro and cryopreservation

At the genebank of IPK (Leibniz Institute of Plant Genetics and Crop Plant Research) in Gatersleben, Germany, three long-term conservation methods (field genebank, in vitro slow-growth and cryopreservation) are used for mint germplasm. The plant material of the field genebank was the source to establish the slow-growth in vitro culture collection, from which the cryopreserved collection was set up, using a droplet-vitrification protocol. The genetic identity of 161 samples of three mint accessions (MEN 198, MEN 166 and MEN 186), stored for several years using those three methods, was studied using RAPD markers. Accession ‘MEN 198’ was the only one with a unique RAPD fragment pattern for all its samples, independently of the conservation procedure employed. The field collections of accessions ‘MEN 166’ and ‘MEN 186’ were made up of different genotypes. None of the genotypes detected in the plots of these accessions was represented in the in vitro and cryopreserved samples analyzed, which showed a unique genotype for each accession. From this work we conclude that, for an appropriate management of a germplasm collection of vegetatively propagated species, the determination of the genetic composition of the donor material and a periodical assessment of the preserved material should be carried out.