Integrating the Use of Spring- and Fall-Calving Beef Cows in a Year-round Grazing System (A Progress Report)

Animal production, hay production and feeding, and the yields and composition of forage from summer and winter grass-legume pastures and winter corn crop residue fields from a year-round grazing system were compared with those of a conventional system. The year-round grazing system utilized 1.67 acres of smooth bromegrass-orchardgrass-birdsfoot trefoil pasture per cow in the summer, and 1.25 acres of stockpiled tall fescue-red clover pasture per cow, 1.25 acres of stockpiled smooth bromegrass-red clover pasture per cow, and 1.25 acres of corn crop residues per cow during winter for spring- and fall-calving cows and stockers. First-cutting hay was harvested from the tall fescue-red clover and smooth bromegrass-red clover pastures to meet supplemental needs of cows and calves during winter. In the conventional system (called the minimal land system), spring-calving cows grazed smooth bromegrass-orchardgrass-birdsfoot trefoil pastures at 3.33 acres/cow during summer with first cutting hay removed from one-half of these acres. This hay was fed to these cows in a drylot during winter. All summer grazing was done by rotational stocking for both systems, and winter grazing of the corn crop residues and stockpiled forages for pregnant spring-calving cows and lactating fall-calving cows in the year-round system was managed by strip-stocking. Hay was fed to springcalving cows in both systems to maintain a mean body condition score of 5 on a 9-point scale, but was fed to fall-calving cows to maintain a mean body condition score of greater than 3. Over winter, fall-calving cows lost more body weight and condition than spring calving cows, but there were no differences in body weight or condition score change between spring-calving cows in either system. Fall- and spring-calving cows in the yearround grazing system required 934 and 1,395 lb. hay dry matter/cow for maintenance during the winter whereas spring-calving cows in drylot required 4,776 lb. hay dry matter/cow. Rebreeding rates were not affected by management system. Average daily gains of spring-born calves did not differ between systems, but were greater than fall calves. Because of differences in land areas for the two systems, weight production of calves per acre of cows in the minimal land system was greater than those of the year-round grazing system, but when the additional weight gains of the stocker cattle were considered, production of total growing animals did not differ between the two systems.

Efficacy of Grazing Stockpiled Perennial Forages for Winter Maintenance of Beef Cows

In a three year study, wintering systems utilizing the grazing of stockpiled perennial hay crop forages or corn crop residues were compared to maintaining cows in a drylot. In the summer of 1992, two cuttings of hay were harvested (June 22 and August 2) from three 10-acre fields containing “Johnstone” endophyte-free tall fescue and “Spreador II” alfalfa, and one cutting of hay was harvested from three 10- acre fields of smooth brome grass. “Arlington” red clover was frost-seeded into the smooth bromegrass fields in 1993 and into tall fescue-alfalfa and smooth bromegrass fields into 1994. Two cuttings of hay were harvested from all fields in subsequent years, and three-year average hay yields for tall fescue-alfalfa and smooth bromegrass-red clover were 4,336 and 3,481 pounds per acre, respectively. Regrowth of the forage following the August hay harvest of each year was accumulated for winter grazing. Following a killing frost in each year, two fields of each stockpiled forage were stocked with cows in midgestation at two acres per cow. Two 10-acre fields of corn crop residues were also stocked at two acres per cow, following the grain harvest. Mean dry matter forage yields at the initiation of grazing were 1,853, 2,173 and 5,797 pounds per acre for fields containing tall fescue-alfalfa, smooth bromegrass-red clover, and cornstalks, respectively. A drylot was stocked with 18 cows in 1992 and 1993 and 10 cows in 1994. All cows were fed hay as necessary to maintain a body condition score of five. During grazing, mean losses of organic matter were -6.4, -7.6, and -10.7 pounds per acre per cow from tall fescue-alfalfa, smooth bromegrass-red clover, and cornstalk fields. Average organic matter loss rates from stockpiled forages due to weathering alone were equal to only 30% of the weathering losses of the corn crop residues. In vitro digestibility of both stockpiled forages and cornstalks decreased at equal rates during grazing each year, with respective annual loss rates of .14, .08, and .06% per day. Cows grazing corn crop residues required an average of 1,321 pounds per cow less hay than cows maintained in the drylot to maintain equivalent body condition during the grazing season. Cows grazing tall fescue-alfalfa or smooth bromegrass-red clover had body weight gains and condition score changes equal to cows maintained in a drylot but required 64% and 62% less harvested hay than cows in the drylot during the grazing season. Over the entire stored forage cows grazing tall fescue-alfalfa and smooth bromegrass-red clover required an average of 2,390 and 2,337 pounds per cow less than those maintained in the drylot. Because less hay was needed to maintain cows grazing stockpiled forages, average annual excesses of 5,629 and 3,868 pounds of hay dry matter per cow remained in the stockpiled tall fescue-alfalfa and smooth bromegrass-red clover systems.

Evaluation of Year-round Forage Management Systems for Spring- and Fall-Calving Beef Cows (A Progress Report)

A year-round grazing system for spring- and fall-calving cows was developed to compare animal production and performance, hay production and feeding, winter forage composition changes, and summer pasture yield and nutrient composition to that from a conventional, or minimal land system. Systems compared forage from smooth bromegrass-orchardgrass-birdsfoot trefoil pastures for both systems in the summer and corn crop residues and stockpiled grass-legume pastures for the year-round system to drylot hay feeding during winter for the minimal land system. The year-round grazing system utilized 1.67 acres of smooth bromegrassorchardgrass- birdsfoot trefoil (SB-O-T) pasture per cow in the summer, compared with 3.33 acres of (SB-O-T) pasture per cow in the control (minimal land) system. In addition to SB-O-T pastures, the year-round grazing system utilized 2.5 acres of tall fescue-red clover (TFRC) and 2.5 acres of smooth bromegrass-red clover (SBRC) per cow for grazing in both mid-summer and winter for fall- and spring-calving cows, respectively. First-cutting hay was harvested from the TF-RC and SB-RC pastures, and regrowth was grazed for approximately 45 days in the summer. These pastures were then fertilized with 40 lbs N/acre and stockpiled for winter grazing. Also utilized during the winter for spring-calving cows in the year-round grazing system were corn crop residue (CCR) pastures at an allowance of 2.5 acres per cow. In the minimal land system, hay was harvested from three-fourths of the area in SB-O-T pastures and stored for feeding in a drylot through the winter. Summer grazing was managed with rotational stocking for both systems, and winter grazing of stockpiled forages and corn crop residues by year-round system cows was managed by strip-stocking. Hay was fed to maintain a body condition score of 5 on a 9 point scale for spring-calving cows in both systems. Hay was supplemented as needed to maintain a body condition score of 3 for fall-calving cows nursing calves through the winter. Although initial condition scores for cows in both systems were different at the initiation of grazing for both winter and summer, there were no significant differences (P > .05) in overall condition score changes throughout both grazing seasons. In year 1, fall-calving cows in the year-round grazing system lost more (P < .05) body weight during winter than spring-calving cows in either system. In year 2, there were no differences seen in weight changes over winter for any group of cows. Average daily gains of fall calves in the yearround system were 1.9 lbs/day compared with weight gains of 2.5 lbs/day for spring calves from both systems. Yearly growing animal production from pastures for both years did not differ between systems when weight gains of stockers that grazed summer pastures in the year-round grazing system were added to weight gains of suckling calves. Carcass characteristics for all calves finished in the feedlot for both systems were similar. There were no significant differences in hay production between systems for year 1; however, amounts of hay needed to maintain cows were 923, 1373, 4732 lbs dry matter/cow for year-round fall-calving, year-round spring-calving, and minimal land spring-calving cows, respectively. In year 2, hay production per acre in the minimal land system was greater (P < .05) than for the year-round system, but the amounts of hay required per cow were 0, 0, and 4720 lbs dry matter/cow for yearround fall-calving, year-round spring-calving, and minimal land spring-calving cows, respectively.

Nutritive Value of the Crop Residues from bt-Corn Hybrids and Their Effects on Performance of Grazing Beef Cows

One non bt-corn hybrid (Pioneer 3489) and three btcorn hyrids (Pioneer 34RO7, Novartis NX6236, and Novartis N64-Z4) were planted in replicated 7.1-acre fields. After grain harvest, fields were stocked with 3 mature cows in midgestation to be strip-grazed as four paddocks over 126 days. Six similar cows were allotted to replicated drylots. All cows were fed hay as necessary to maintain a condition score of 5 on a 9-point scale. Cows were condition-scored biweekly and weighed monthly. Forage yield and weathering losses were determined by sampling one 4-m2 location per grazed or ungrazed paddock in each field with a minimum total of 2 locations of grazed or ungrazed forage per field. To measure forage selection during grazing, samples of grazed forage were collected from the rumen of one fistulated steer that grazed for 2 hours after ruminal evacuation. Non-bt-corn hybrids had greater (P<.05) infestation of corn borers in the upper stalk, lower stalk and ear shank than bt-corn hybrids. However, there were no differences in grain yields or dropped grain between hybrids. Crop residue dry matter, organic matter and in vitro digestible dry matter yields at the initiation of grazing did not differ between corn hybrids. Dry matter, organic matter and in vitro digestible dry matter losses tended (P<.10) to be greater from the NX6236 and N64-Z4 hybrids than from the 3489 and 34RO7 hybrids and were greater (P<.05) from grazed than non-grazed areas of the fields. At the initiation of grazing, dry matter concentrations of the crop residues from the NX6236 and N64-Z4 hybrids tended to be lower than those from the 3489 and 34RO7 hybrids. Crop residues from the NX6236 and N64-74 hybrids had lower concentrations of acid detergent fiber (P<.05) and acid detergent lignin (P=.07) and higher concentrations of in vitro digestible organic matter than the 3489 and 34RO7 hybrids. Over the grazing season, corn hybrid did not affect mean rates of change in forage composition. The concentration of in vitro digestible organic matter in forage selected by steers after two weeks of grazing did not differ. However, steers grazing corn crop residues consumed forage with higher (P<.05) concentrations of neutral detergent fiber, acid detergent fiber, and acid detergent insoluble nitrogen than steers fed hay. The acid detergent fiber concentration of forage selected by steers grazing the 3489 and N64-Z4 hybrids was lower (P < .05) than concentrations from the 34RO7 and NX6236 hybrids. In order to maintain similar body condition score changes, cows grazing crop residues from the 3489, 34RO7, NX6236, and N64-Z4 hybrids required 650, 628, 625, and 541 kg hay DM/cow compared with a hay requirement of 1447 kg hay DM/cow for cows maintained in a drylot.

Evaluation of Stockpiled Berseem Clover and Brown Midrib Sorghum x Sudangrass as Supplements for Grazed Cornstalks in Beef Cow Wintering Systems

Berseem clover and oats were incorporated into a corncorn- oat/berseem clover rotation in 1994 and 1995. Two cuttings of oat-berseem clover hay were harvested during the summer before forage was allowed to stockpile for winter grazing. In 1995, a brown midrib sorghum x sudangrass hybrid was seeded into a field adjacent to a corn field. After corn grain harvest in 1994 and 1995, Charolais x Angus x Simmental cows in midgestation were allotted to replicated fields containing corn crop residues with no complementary forages at 2.5 acres/cow, or corn crop residues and stockpiled berseem clover (2:1) at 2.5 acres/cow to simultaneously graze, or to a drylot. In 1995, cows were allotted to fields containing corn crop residues and brown midrib sorghum x sudangrass (7:3) at 2.5 acres/cow. Berseem clover had greater concentrations of digestible organic matter and crude protein than corn crop residues at the initiation of grazing, but had a more rapid decrease in digestible organic matter concentration than corn crop residues. Brown midrib sorghum x sudangrass forage also had a higher initial concentration of digestible organic matter, but an equal rate of decrease in digestible organic matter concentration to corn crop residues in ungrazed areas of the field. Cows grazing berseem clover with corn crop residues had greater body condition score increases during the first half of the grazing season than cows grazing corn crop residues without complementary forages. Cows grazing corn crop residues without complementary forages required 2,786 and 1,412 less lb hay per cow than cows maintained in a drylot in 1994 and 1995. In 1994, simultaneous grazing of berseem clover with corn crop residues did not reduce hay feeding more than feeding corn crop residues alone. However, in 1995, grazing berseem clover or brown midrib sorghum x sudangrass with corn crop residues reduced the amount of hay required to maintain cows by 358 and 376 lb hay per cow compared with grazing corn crop residues without complementary forage.

Evaluation of Stockpiled Berseem Clover and Brown Midrib Sorghum x Sudangrass as Supplements for Grazed Cornstalks in Beef Cow Wintering Systems

Berseem clover and oats were incorporated into a corn-oat/berseem clover rotation in 1994-1996. Two cuttings of oat-berseem clover hay were harvested during the summer before forage was stockpiled for winter grazing. In 1995, brown midrib sorghum x sudangrass hybrid was seeded into a field adjacent to a corn field. This was repeated in 1996 with a standard sorghum x sudangrass hybrid. After corn harvest in 1994–1996, Charolais x Angus x Simmental cows and heifers in midgestation were allotted to corn crop residue, corn crop residue-berseem clover, and corn crop residue-sorghum x sudangrass fields at 2.5 acres/cow, or to a drylot. Berseem clover had greater concentration of digestible organic matter and crude protein than corn crop residues. Corn crop residue digestible organic matter concentration was lower than berseem clover and the brown midrib sorghum x sudangrass, but was higher than that of the standard sorghum x sudangrass hybrid in 1996. Cows grazing corn crop residues without complementary forages required an average of 2,374 less lb. hay per cow than cows maintained in a drylot in 1994-1996. In 1994 and 1996, simultaneous grazing of berseem clover with corn crop residues did not reduce hay feeding more than feeding corn crop residues alone, yet did significantly reduce the amount of hay needed in 1995 to maintain cows by 358 and 376 lb. hay per cow compared with grazing corn crop residues without complementary forage.

The Effect of Heat Treatment of Forages on Degradation Kinetics and Escape Protein Concentration

An in situ study was conducted to evaluate the effects of heat treatments on the degradation kinetics and escape protein concentrations of forages (alfalfa and berseem clover). Alfalfa collected at 4 and 7 weeks post-harvest and berseem clover collected at 5 and 7 weeks postharvest were freeze-dried and then heated to 100, 125, and 150o C for 2 hours. Heat treatment effects were determined by placing two bags of sample (for each treatment, maturity, and forage species for a given incubation times) into the rumen of one fistulated steer fed alfalfa hay. Bags were incubated for periods of 0 to 48 hours. Increasing levels of heat treatments of forages increased concentrations of neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent insoluble nitrogen (ADIN) and non-degradable protein (NDP), potentially degradable protein proportion (PDP), and protein escaping rumen degradation (PEP) while decreasing water soluble protein (WSP) and the rates of crude protein (CP), except immature berseem clover and cell wall (CW) degradation. PEP was greater and rate of CP degradation was lower at 100 and 150o C compared to 125o C in immature berseem clover.

Rapid and simple LC-MS/MS-Screening of 64 novel psychoactive substances using dried blood spots

The range of novel psychoactive substances (NPS) including phenethylamines, cathinones, piperazines, tryptamines, etc. is continuously growing. Therefore, fast and reliable screening methods for these compounds are essential and needed. The use of dried blood spots (DBS) for a fast straightforward approach helps to simplify and shorten sample preparation significantly. DBS were produced from 10 µl of whole blood and extracted offline with 500 µl methanol followed by evaporation and reconstitution in mobile phase. Reversed-phase chromatographic separation and mass spectrometric detection (RP-LC-MS/MS) was achieved within a run time of 10 min. The screening method was validated by evaluating the following parameters: limit of detection (LOD), matrix effect, selectivity and specificity, extraction efficiency, and short-term and long-term stability. Furthermore, the method was applied to authentic samples and results were compared with those obtained with a validated whole blood method used for Routine analysis of NPS. LOD was between 1 and 10 ng/ml. No interference from Matrix compounds was observed. The method was proven to be specific and selective for the analytes, although with limitations for 3-FMC/flephedrone and MDDMA/MDEA. Mean extraction efficiency was 84.6 %. All substances were stable in DBS for at least a week when cooled. Cooling was essential for the stability of cathinones. Prepared samples were stable for at least 3 days. Comparison to the validated whole blood method yielded similar results. DBS were shown to be useful in developing a rapid screening method for NPS with simplified sample preparation. Copyright © 2013 John Wiley & Sons, Ltd

Determination of Ethyl glucuronide and Ethyl sulfate from dried blood spots

BACKGROUND: Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are non-oxidative minor metabolites of ethanol. They are detectable in various body fluids shortly after initial consumption of ethanol and have a longer detection time frame than the parent compound. They are regarded highly sensitive and specific markers of recent alcohol uptake. This study evaluates the determination of EtG and EtS from dried blood spots (DBS), a simple and cost-effective sampling method that would shorten the time gap between offense and blood sampling and lead to a better reflectance of the actual impairment. METHODS: For method validation, EtG and EtS standard and quality control samples were prepared in fresh human heparinized blood and spotted on DBS cards, then extracted and measured by an LC-ESI-MS/MS method. Additionally, 76 heparinized blood samples from traffic offense cases were analyzed for EtG and EtS as whole blood and as DBS specimens. The results from these measurements were then compared by calculating the respective mean values, by a matched-paired t test, by a Wilcoxon test, and by Bland-Altman and Mountain plots. RESULTS AND DISCUSSION: Calibrations for EtG and EtS in DBS were linear over the studied calibration range. The precision and accuracy of the method met the requirements of the validation guidelines that were employed in the study. The stability of the biomarkers stored as DBS was demonstrated under different storage conditions. The t test showed no significant difference between whole blood and DBS in the determination of EtG and EtS. In addition, the Bland-Altman analysis and Mountain plot confirmed that the concentration differences that were measured in DBS specimens were not relevant.

The basel cocktail for simultaneous phenotyping of human cytochrome P450 isoforms in plasma, saliva and dried blood spots.

BACKGROUND AND OBJECTIVE Phenotyping cocktails use a combination of cytochrome P450 (CYP)-specific probe drugs to simultaneously assess the activity of different CYP isoforms. To improve the clinical applicability of CYP phenotyping, the main objectives of this study were to develop a new cocktail based on probe drugs that are widely used in clinical practice and to test whether alternative sampling methods such as collection of dried blood spots (DBS) or saliva could be used to simplify the sampling process. METHODS In a randomized crossover study, a new combination of commercially available probe drugs (the Basel cocktail) was tested for simultaneous phenotyping of CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. Sixteen subjects received low doses of caffeine, efavirenz, losartan, omeprazole, metoprolol and midazolam in different combinations. All subjects were genotyped, and full pharmacokinetic profiles of the probe drugs and their main metabolites were determined in plasma, dried blood spots and saliva samples. RESULTS The Basel cocktail was well tolerated, and bioequivalence tests showed no evidence of mutual interactions between the probe drugs. In plasma, single timepoint metabolic ratios at 2 h (for CYP2C19 and CYP3A4) or at 8 h (for the other isoforms) after dosing showed high correlations with corresponding area under the concentration-time curve (AUC) ratios (AUC0-24h parent/AUC0-24h metabolite) and are proposed as simple phenotyping metrics. Metabolic ratios in dried blood spots (for CYP1A2 and CYP2C19) or in saliva samples (for CYP1A2) were comparable to plasma ratios and offer the option of minimally invasive or non-invasive phenotyping of these isoforms. CONCLUSIONS This new combination of phenotyping probe drugs can be used without mutual interactions. The proposed sampling timepoints have the potential to facilitate clinical application of phenotyping but require further validation in conditions of altered CYP activity. The use of DBS or saliva samples seems feasible for phenotyping of the selected CYP isoforms.

Observed management practices in relation to the risk of infection with paratuberculosis and to the spread of Mycobacterium avium subsp. paratuberculosis in Swiss dairy and beef herds

Abstract BACKGROUND: Many studies have been conducted to define risk factors for the transmission of bovine paratuberculosis, mostly in countries with large herds. Little is known about the epidemiology in infected Swiss herds and risk factors important for transmission in smaller herds. Therefore, the presence of known factors which might favor the spread of paratuberculosis and could be related to the prevalence at animal level of fecal shedding of Mycobacterium avium subsp. paratuberculosis were assessed in 17 infected herds (10 dairy, 7 beef). Additionally, the level of knowledge of herd managers about the disease was assessed. In a case-control study with 4 matched negative control herds per infected herd, the association of potential risk factors with the infection status of the herd was investigated. RESULTS: Exposure of the young stock to feces of older animals was frequently observed in infected and in control herds. The farmers' knowledge about paratuberculosis was very limited, even in infected herds. An overall prevalence at animal level of fecal shedding of Mycobacterium avium subsp. paratuberculosis of 6.1% was found in infected herds, whereby shedders younger than 2 years of age were found in 46.2% of the herds where the young stock was available for testing. Several factors related to contamination of the heifer area with cows' feces and the management of the calving area were found to be significantly associated with the within-herd prevalence. Animal purchase was associated with a positive herd infection status (OR = 7.25, p = 0.004). CONCLUSIONS: Numerous risk factors favoring the spread of Mycobacterium avium subsp. paratuberculosis from adult animals to the young stock were observed in infected Swiss dairy and beef herds, which may be amenable to improvement in order to control the disease. Important factors were contamination of the heifer and the calving area, which were associated with higher within-herd prevalence of fecal shedding. The awareness of farmers of paratuberculosis was very low, even in infected herds. Animal purchase in a herd was significantly associated with the probability of a herd to be infected and is thus the most important factor for the control of the spread of disease between farms.

Conditioned Medium of Demineralized Freeze-Dried Bone Activates Gene Expression in Periodontal Fibroblasts in Vitro.

BACKGROUND: Demineralized bone matrix (DBM) is used for the treatment of osseous defects. Conditioned medium from native bone chips can activate transforming growth factor (TGF)-β signaling in mesenchymal cells. The aim of the study was to determine whether processing of native bone into DBM affects the activity of the conditioned medium. METHODS: Porcine cortical bone blocks were subjected to defatting, different concentrations of hydrochloric acid and various temperatures. DBM was lyophilized, ground, and placed into culture medium. Human gingiva and periodontal fibroblasts were exposed to the respective conditioned medium (DBCM). Changes in the expression of TGF-β target genes were determined. RESULTS: DBCM altered the expression of TGF-β target genes, e.g., adrenomedullin, pentraxin 3, KN Motif And Ankyrin Repeat Domains 4, interleukin 11, NADPH oxidase 4, and BTB (POZ) Domain Containing 11, by at least five-fold. The response was observed in fibroblasts from both sources. Defatting lowered the activity of DBCM. The TGF-β receptor type I kinase inhibitor SB431542, but not the inhibitor of bone morphogenetic protein receptor dorsomorphin, blocked the effects of DBCM on gene expression. Moreover, conditioned medium obtained from commercial human DBM modulated the expression of TGF-β target genes. CONCLUSION: The findings suggest that the conditioned medium from demineralized bone matrix can activate TGF-β signaling in oral fibroblasts. KEYWORDS: TGF-beta superfamily proteins; bone; bone substitutes; bone transplantation; conditioned media; freeze drying

Hepatitis B viral load in dried blood spots: A validation study in Zambia

Background: Access to hepatitis B viral load (VL) testing is poor in sub-Saharan Africa (SSA) due toeconomic and logistical reasons.Objectives: To demonstrate the feasibility of testing dried blood spots (DBS) for hepatitis B virus (HBV)VL in a laboratory in Lusaka, Zambia, and to compare HBV VLs between DBS and plasma samples.Study design: Paired plasma and DBS samples from HIV-HBV co-infected Zambian adults were analyzedfor HBV VL using the COBAS AmpliPrep/COBAS TaqMan HBV test (Version 2.0) and for HBV genotypeby direct sequencing. We used Bland-Altman analysis to compare VLs between sample types and bygenotype. Logistic regression analysis was conducted to assess the probability of an undetectable DBSresult by plasma VL.Results: Among 68 participants, median age was 34 years, 61.8% were men, and median plasma HBV VLwas 3.98 log IU/ml (interquartile range, 2.04–5.95). Among sequenced viruses, 28 were genotype A1 and27 were genotype E. Bland–Altman plots suggested strong agreement between DBS and plasma VLs. DBSVLs were on average 1.59 log IU/ml lower than plasma with 95% limits of agreement of −2.40 to −0.83 logIU/ml. At a plasma VL ≥2,000 IU/ml, the probability of an undetectable DBS result was 1.8% (95% CI:0.5–6.6). At plasma VL ≥20,000 IU/ml this probability reduced to 0.2% (95% CI: 0.03–1.7).

Determination of fatty acid ethyl esters in dried blood spots by LC-MS/MS as markers for ethanol intake: application in a drinking study.

The forensic utility of fatty acid ethyl esters (FAEEs) in dried blood spots (DBS) as short-term confirmatory markers for ethanol intake was examined. An LC-MS/MS method for the determination of FAEEs in DBS was developed and validated to investigate FAEE formation and elimination in a drinking study, whereby eight subjects ingested 0.66-0.84 g/kg alcohol to reach blood alcohol concentrations (BAC) of 0.8 g/kg. Blood was taken every 1.5-2 h, BAC was determined, and dried blood spots were prepared, with 50 μL of blood, for the determination of FAEEs. Lower limits of quantitation (LLOQ) were between 15 and 37 ng/mL for the four major FAEEs. Validation data are presented in detail. In the drinking study, ethyl palmitate and ethyl oleate proved to be the two most suitable markers for FAEE determination. Maximum FAEE concentrations were reached in samples taken 2 or 4 h after the start of drinking. The following mean peak concentrations (c̅ max) were reached: ethyl myristate 14 ± 4 ng/mL, ethyl palmitate 144 ± 35 ng/mL, ethyl oleate 125 ± 55 ng/mL, ethyl stearate 71 ± 21 ng/mL, total FAEEs 344 ± 91 ng/mL. Detectability of FAEEs was found to be on the same time scale as BAC. In liquid blood samples containing ethanol, FAEE concentrations increase post-sampling. This study shows that the use of DBS fixation prevents additional FAEE formation in blood samples containing ethanol. Positive FAEE results obtained by DBS analysis can be used as evidence for the presence of ethanol in the original blood sample. Graphical Abstract Time courses for fatty acid ethyl ester (FAEE) concentrations in DBS and ethanol concentrations for subject 1 over a period of 7 h. Ethanol ingestion occured during the first hour of the time course.

Field investigation of dried lakes in western United States as an analogue to desiccation fractures on Mars

Potential Desiccation Polygons (PDPs), tens to hundreds of meters in size, have been observed in numerous regions on Mars, particularly in ancient (>3Gyr old) terrains of inferred paleolacustrine/playa geologic setting, and in association with hydrous minerals such as smectites. Therefore, a better understanding of the conditions in which large desiccation polygons form could yield unique insight into the ancient climate on Mars. Many dried lakebeds/playas in western United States display large (>50m wide) desiccation polygons, which we consider to be analogues for PDPs on Mars. Therefore, we have carried out fieldwork in seven of these dried lakes in San Bernardino and the Death Valley National Park regions complemented with laboratory and spectral analysis of collected samples. Our study shows that the investigated lacustrine/playa sediments have (a) a soil matrix containing 40-75% clays and fine silt (by volume) where the clay minerals are dominated by illite/muscovite followed by smectite, (b) carbonaceous mineralogy with variable amounts of chloride and sulfate salts, and significantly, (c) roughly similar spectral signatures in the visible-near-infrared (VIS-NIR) range. We conclude that the development of large desiccation fractures is consistent with water table retreat. In addition, the comparison of the mineralogical to the spectral observations further suggests that remote sensing VIS-NIR spectroscopy has its limitations for detailed characterization of lacustrine/playa deposits. Finally, our results imply that the widespread distribution of PDPs on Mars indicates global or regional climatic transitions from wet conditions to more arid ones making them important candidate sites for future in situ missions.