On the Habitability of Desert Varnish: A Combined Study by Micro-Raman Spectroscopy, X-ray Diffraction, and Methylated Pyrolysis-Gas Chromatography-Mass Spectrometry.

In 2020, the ESA ExoMars and NASA Mars 2020 missions will be launched to Mars to search for evidence of past and present life. In preparation for these missions, terrestrial analog samples of rock formations on Mars are studied in detail in order to optimize the scientific information that the analytical instrumentation will return. Desert varnishes are thin mineral coatings found on rocks in arid and semi-arid environments on Earth that are recognized as analog samples. During the formation of desert varnishes (which takes many hundreds of years), organic matter is incorporated, and microorganisms may also play an active role in the formation process. During this study, four complementary analytical techniques proposed for Mars missions (X-ray diffraction [XRD], Raman spectroscopy, elemental analysis, and pyrolysis-gas chromatography-mass spectrometry [Py-GC-MS]) were used to interrogate samples of desert varnish and describe their capacity to sustain life under extreme scenarios. For the first time, both the geochemistry and the organic compounds associated with desert varnish are described with the use of identical sets of samples. XRD and Raman spectroscopy measurements were used to nondestructively interrogate the mineralogy of the samples. In addition, the use of Raman spectroscopy instruments enabled the detection of ß-carotene, a highly Raman-active biomarker. The content and the nature of the organic material in the samples were further investigated with elemental analysis and methylated Py-GC-MS, and a bacterial origin was determined to be likely. In the context of planetary exploration, we describe the habitable nature of desert varnish based on the biogeochemical composition of the samples. Possible interference of the geological substrate on the detectability of pyrolysis products is also suggested. Key Words: Desert varnish-Habitability-Raman spectroscopy-Py-GC-MS-XRD-ExoMars-Planetary science. Astrobiology 17, 1123-1137.

Interactions between oil substrates and glucose on pure cultures of ruminal lipase-producing bacteria.

The hydrolysis of free fatty acids from lipids is a prerequisite for biohydrogenation, a process that effectively saturates free fatty acids. Anaerovibrio lipolyticus 5s and Butyrivibrio fibrisolvens have long been thought to be the major contributors to ruminal lipolysis; however, Propionibacterium avidum and acnes recently have been identified as contributing lipase activity in the rumen. In order to further characterize the lipase activity of these bacterial populations, each was grown with three different lipid substrates, olive oil, corn oil, and flaxseed oil (3 %). Because different finishing rations contain varying levels of glycogen (a source of free glucose) this study also documented the effects of glucose on lipolysis. P. avidum and A. lipolyticus 5s demonstrated the most rapid rates (P < 0.05) of lipolysis for cultures grown with olive oil and flaxseed oil, respectively. A. lipolyticus, B. fibrisolvens, and P. avidum more effectively hydrolyzed flaxseed oil than olive oil or corn oil, especially in the presence of 0.02 % glucose. Conversely, P. acnes hydrolyzed corn oil more readily than olive oil or flaxseed oil and glucose had no effect on lipolytic rate. Thus, these bacterial species demonstrated different specificities for oil substrates and different sensitivities to glucose.

Experimental approach to optimize phytate phosphorus utilization by broiler chickens by addition of supplements.

Supplementation of 1alpha-hydroxycholecal-ciferol (1alpha-OHD(3)), phytase, and Ca levels has been shown to influence phytate P utilization. Two experiments were conducted with 1- to 16-d broilers to investigate the interactions between the effects of 1alpha-OHD(3), phytase, and Ca on phytate P utilization. In experiment 1, the birds were randomly allocated to 13 treatments with 6 levels of 1alpha-OHD(3) from 0 to 11microg/kg, with and without 500 U/kg of phytase supplemented to P-deficient diet. A positive control diet, adequate in Ca and P, was included. In experiment 2, a 3-dimensional rotatable design with 5 levels each of 1alpha-OHD(3), phytase, and Ca was implemented. Supplementation of 1alpha-OHD(3) and phytase increased phytate P utilization, as indicated by an increase in bone ash, phytate P disappearance, and BW. In experiment 1, no interaction between the 1alpha-OHD(3) and phytase effects was determined for any criteria, except for the incidence of P rickets. However, the second experiment showed that there were interactions between the 3 factors (1alpha-OHD(3), phytase, and Ca). The probabilities that observed variation in the 3-way interaction for measured criteria that were due to chance are: BW gain = 0.1364, G:F = 0.11, bone ash percentage = 0.0278, milligrams of bone ash/tibia = 0.0909, Ca rickets = 0.9394, P rickets = 0.4892, tibial dyschondroplasia = 0.5927, and phytate P disappearance = 0.1126. The equations obtained from the regression models were used to generate contour-surface plots. The percentage of bone ash data indicated that low Ca, high phytase, and high 1alpha-OHD(3) levels were needed to optimize phytate P utilization. The results of this study show that this experimental design may be utilized to identify the best levels of nutrients to add to a diet where complex 3-way interactions exist affecting several criteria. The equations may be used to optimize performance based on desired levels of performance and the costs of the inputs.

Romano-British wall paintings: Raman spectroscopic analysis of fragments from two urban sites of early military colonisation.

Raman spectroscopic analyses of 1st century AD Romano-British villa wall-painting fragments from two important military and early urban centres at Colchester and Lincoln have demonstrated some interesting contrasts in technique and palette usage. Colchester, the earliest fortified settlement, developed a sophisticated painting and craft industry compared with Lincoln in the assimilation of novel substrate preparation ideas and pigment adoption. The earliest use of the rather rare purple mineral pigment, caput mortuum, hitherto reported in only a few Roman villas elsewhere in mainland Europe, is in evidence in this early phase settlement and the use of gypsum as a special ground preparation agent as an additive to the more common limewash putty to enhance the effect of the use of lazurite as a pigment is worthy of note in this context. Otherwise, the pigments are seen to be those that are quite normally encountered in Roman villas, namely, haematite, goethite, terre verte, and carbon. The results of this study indicate that at Colchester there was a continued development in technique into the colonial phase compared with a stagnation in Lincoln; these scientific results have created a stimulus for further historical research into pigment and techniques development for wall paintings at the fringe of the Roman Empire in the 1st-3rd Centuries AD.

The effect of breeder age and egg storage time on phosphorus utilization by broiler progeny fed a phosphorus deficiency diet with 1alpha-OH vitamin D3.

Two experiments were conducted to determine that variation in broiler P utilization is due to breeder age and egg storage time. Experiment 1 was conducted with chicks hatched from eggs laid by Ross x Ross 308 breeders (27 vs. 61 wk old) and stored for 0 or 10 d. The age of breeders had significant effects (P < 0.05) on 0 to 16 d chick growth (379 +/- 18 vs. 308 +/- 19 for 27- and 61-wk-old breeders, respectively). The longer egg storage time of chicks from older breeders resulted in higher P rickets scores and incidence, but longer egg storage time of chicks from younger breeders resulted in lower P rickets score and incidence (significant interaction, P = 0.0455). The longer egg storage time of chicks from older breeders resulted in lower bone ash (%), and the longer egg storage time of chicks from younger breeders resulted in higher bone ash (%). Experiment 2 was conducted with chicks hatched from eggs laid by Ross x Ross 308 breeders (26 vs. 60 wk old) and stored for 0 or 10 d. The diets were P deficient and with or without 5 microg/g of 1alpha-OH cholecalciferol (1alpha-OH vitamin D(3)). Breeder age had significant effects (P = 0.0003) on 0 to 16 d chick growth (272 +/- 7 vs. 339 +/- 8 for 26- and 60-wk-old breeders, respectively) and chick mortality (P = 0.0134). The P rickets score increased with breeder age (P = 0.0186) and egg storage time (P = 0.1057). The factors influencing the incidence of P rickets in broilers should include breeder age and egg storage time as well as genetics and dietary levels of Ca, P, and vitamin D activity of the P-deficient diets.

The effect of several organic acids on phytate phosphorus hydrolysis in broiler chicks.

Supplementation of some organic acids to a P-deficient diet has been shown to improve phytate P utilization. Two experiments were conducted from 0 to 16 d in battery brooders to determine the effect of various organic acids supplementation on phytate P utilization. In both experiments, birds were fed P-deficient corn and soybean meal-based diets. In experiment 1, citric acid, malic acid, fumaric acid, and EDTA were supplemented. Experiment 2 had a 2 x 2 factorial design with 2 sources of Met, 2-hydroxy-4-(methylthio) butanoic acid (HMB) and dl-Met, with or without 500 U/kg of phytase. In experiment 1, the addition of citric, malic, and fumaric acids increased percentage of bone ash, but only the effect of citric acid was significant. The addition of citric and malic acids also significantly increased the retention of P and phytate P (P<0.05). In experiment 2, the addition of phytase to the diet significantly increased 16-d BW gain, feed intake, percentage of bone ash, milligrams of bone ash, phytate P disappearance, and decreased the incidence of P-deficiency rickets. Methionine source did not affect 16-d BW gain, feed intake, feed efficiency, milligrams of bone ash, or P rickets incidence. However, the birds fed HMB had a higher percentage of bone ash and phytate P disappearance compared with the groups fed dl-Met only when phytase was added to the diets. The additions of citric acid and HMB improved phytate P utilization. However, the reason why some organic acids are effective whereas others are not is not apparent.

Analytical discrimination between sources of ginseng using Raman spectroscopy.

Ginseng is a widely used medicinal product that grows mainly in Korea, China and America. American ginseng is classified as an endangered species, and so the import and export of this type of ginseng is illegal in certain countries. Due to this restriction it is becoming increasingly important to be able to distinguish between different types of ginseng. FT-Raman spectroscopy has the ability to discriminate between ginseng specimens according to the country of origin and the effects of processing on the ginseng material. The ginsenoside content of ginseng differs in both conformation and concentration depending on the source of the ginseng, which means that ginseng grown in different countries should express unique spectral features. The presence or absence of these features, therefore, could indicate the geographical origin of the sample. Several spectral features were identified for a range of ginsengs, such as a peak at 980 cm(-1) that was only found in Chinese ginseng, and the different wavenumber positions of characteristic ginseng bands near 1600 cm(-1). This indicates that Raman spectroscopy can be used to pinpoint the origin of an unknown ginseng sample and that it would provide a rapid nondestructive analytical technique for formally discriminating between restricted and permitted imports.

The effect of feeding calcium- and phosphorus-deficient diets to broiler chickens during the starting and growing-finishing phases on carcass quality.

There is considerable data on the effect of reducing inorganic Ca and P in broiler finisher diets on carcass quality. However, there is limited information on the effect of reducing dietary Ca and P during the different phases of growout. Two experiments were conducted from 0 to 35 d in floor pens. In both experiments, at least 4 replicates per treatment (50 chicks per replicate) were used. Corn-soybean meal and soybean oil-based diets deficient in Ca and P were fed. During the starter phase (ST), from 0 to 18 d, chicks were fed a 23% CP diet containing 0.60% Ca and 0.47% total P (tP). During the grower-finisher phase (GF), from 19 to 35 d, birds were fed a 19% CP diet containing 0.30% Ca and 0.37% tP. A combination of 1,000 phytase units/kg of Natuphos phytase and 5 microg/kg of 1alpha-hydroxycholecalciferol (P + 1alpha) was supplemented to some of the feed during the ST and GF. Diets containing adequate Ca and P were also fed during the ST (0.90% Ca and 0.68% tP) and GF (0.80% Ca and 0.67% tP). The level of tibia ash and the incidence of bone disease were measured at 18 and 35 d. At the end of the experiments, birds were processed and evaluated for muscle hemorrhages and broken bones. In both experiments, broilers fed diets that were not P + 1alpha supplemented demonstrated poor bone mineralization, considerable leg problems, and a high incidence of broken bones after processing. Broilers fed P + 1alpha throughout had more broken clavicles and femurs compared with birds fed the adequate diets. Day-18 tibia ash was significantly correlated to broken tibias and femurs during processing. Day-35 tibia ash was better correlated to bloody breast meat than to broken bones. It is concluded that carcass quality depends on the levels of Ca and P fed and the age of the bird. Tibia ash, traditionally used as an indication of bone strength, was better correlated to the incidence of bloody breasts.

The vitamin D3 requirement of broiler breeders.

An experiment was conducted with 25- to 66-wk-old Ross broiler breeders in an environment excluding ultraviolet light to determine the cholecalciferol (D3) requirements for hen day egg production; hatchability; body weight of the progeny at 1 d; embryo mortality during the early (1 to 10 d of incubation), middle (11 to 15 d of incubation), and late stages (16 to 21 d of incubation) of development; egg weight; specific gravity; and body ash of the progeny at 1 d of age. Five levels of vitamin D3 (125, 250, 500, 1,000, and 2,000 IU/kg of diet) were fed to hens from 25 to 66 wk of age. One additional group was fed no supplemental D3 until 36 wk of age and was then changed to 4,000 IU/kg of diet. Separate regression analyses were performed for wk 27 to 36 (peak original design) and for wk 37 to 66 (postpeak production modified design). The D3 levels for the predicted maximum hen day egg production during peak and postpeak were 1,424 and 2,804 IU/kg, respectively. The D3 levels for the predicted maximum hatchability were 1,390 IU/ kg (peak) and 2,708 IU/kg (postpeak). The level of D3 that resulted in the minimum early embryo mortality was 1,288 IU/kg at peak; however, no significant effect was observed at postpeak. The D3 levels for minimum middle stage embryo mortality were 1,130 IU/kg (peak) and 2,568 IU/kg (postpeak) and for late stage embryo mortality were 1,393 IU/kg (peak) and 2,756 IU/kg (postpeak). The D3 level for maximum egg weight was 1,182 IU/kg (peak) and for specific gravity was 1,337 IU/kg (peak) and >2,000 IU/kg (postpeak). The D3 level for maximum body ash of progeny at d 1 was >2,000 IU/kg. Analysis of the data from the original design of the experiment (treatments providing 0, 125, 250, 500, 1,000, and 2,000 IU of vitamin D3/kg for the 27- to 36-wk-old birds) indicates a requirement of approximately 1,400 IU of D3/kg of feed for broiler breeder hens. When the data from the modified experiment (37 to 66 wk of age) include conversion of the treatment provided at 0 IU of D3/kg to a treatment providing 4,000 IU of D3/kg, the requirement may be approximately 2,800 IU of D3/kg.

The effect of maternal dietary vitamin D3 supplementation on performance and tibial dyschondroplasia of broiler chicks.

A series of experiments was conducted to investigate the effects of maternal dietary vitamin D3 supplementation at 4 different times during the laying cycle, on the performance and bone quality of broiler chicks fed a diet that induced tibial dyschondroplasia (TD) or an adequate diet. Ross x Ross broiler breeder hens were fed a corn-soy diet with various levels of vitamin D3 from 24 to 66 wk of age. Eggs were collected at 39, 44, 53, and 64 wk of age and hatched. Chicks from hens fed 250 IU of D3/kg (low maternal D3 or LMD3) and 2,000 IU of D3/ kg (high maternal D3 or HMD3) levels were placed in battery brooders and fed the diets from 0 to 16 d. At 16 d, the chicks were weighed and killed; the left tibias were used for bone ash determinations, and the right tibias were used to score the incidence and severity of TD (0, 1, 2, or 3, where 3 is the most severe). Body weight gain and feed intake were significantly lower for the LMD3 chicks at wk 44 and 64, although there was no difference in weight at hatch. For the first 2 hatches (wk 39 and 44), the LMD3 and HMD3 chicks demonstrated high average TD scores (2.03 and 1.57 vs. 2.05 and 1.75 for the LMD3 vs. HMD3 chicks, respectively) and high average incidences of severe TD (50 and 35% vs. 45 and 34% for LMD3 vs. HMD3 levels, respectively). However, results from the last 2 hatches (wk 53 and 64) showed that HMD3 chicks, compared with LMD3 chicks, had reduced average TD scores (1.39 and 1.47 vs. 1.01 and 0.44 for LMD3 vs. HMD3 levels, respectively) and severe TD incidence (36 and 40% vs. 17 and 8% for the LMD3 vs. HMD3 levels, respectively). In this experiment, as egg production declined toward the end of the laying cycle, hens fed the HMD3 might have been able to deposit sufficient quantities of vitamin D3 in the egg to maintain excellent body weight gain at 16 d of age and reduce the incidence and severity of TD. Hens fed the LMD3 diet were unable to produce similar improvements.