On-line gas chromatography combustion/pyrolysis isotope ratio mass spectrometry (HRGC-C/P-IRMS) of pineapple (Ananas comosus L. Merr.) volatiles

By use of extracts prepared by liquid-liquid separation of the volatiles from self-prepared juices of pineapple fruits (Ananas comosus) (n = 14) as well as commercial pineapple recovery aromas/water phases (n = 3), on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and the pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta(13)C(VPDB) and delta(2)H(VSMOW) values of selected pineapple flavor constituents. In addition to methyl 2-methylbutanoate 1, ethyl 2-methylbutanoate 2, methyl hexanoate 3, ethyl hexanoate 4, and 2,5-dimethyl-4-methoxy-3[2H]-furanone 5, each originating from the fruit, the delta(13)C(VPDB) and delta(2)H(VSMOW) data of commercial synthetic 1-5 and "natural" (biotechnologically derived) 1-4 were determined. With delta(13)C(VPDB) data of pineapple volatiles 1-4 varying from -12.8 to -24.4 per thousand, the range expected for CAM metabolism was observed. Compound 5 showed higher depletion from -20.9 to -28.6 per thousand. A similar situation was given for the delta(2)H(VSMOW) values of 3-5 from pineapple ranging from -118 to -191 per thousand, whereas 1 and 2 showed higher depleted values from -184 to -263 per thousand. In nearly all cases, analytical differentiation of 1-5 from pineapple and natural as well as synthetic origin was possible. In general, natural and synthetic 1-5 exhibited delta(13)C(VPDB) data ranging from -11.8 to -32.2 per thousand and -22.7 to -35.9 per thousand, respectively. Their delta(2)H(VSMOW) data were in the range from -242 to -323 per thousand and -49 to -163 per thousand, respectively.     

Modelling of soil nutrient budgets: an assessment of agricultural sustainability in Nepal

Abstract. Sustaining soil fertility under agricultural intensification and expansion onto marginal lands is a significant challenge in the Nepalese Middle Mountains. In a detailed watershed study it was shown that the overall soil fertility is poor, forest soils display the poorest conditions as a result of biomass removal, and sustaining agriculture is questionable due to the transformation from traditional to multiple cropping systems. Parent material is a significant factor influencing low phosphorus status while insufficient inputs create deficiencies in total carbon, nitrogen and bases. A nutrient budget model was developed to assess inputs, redistribution and losses relative to soil fertility. Yield, input and management data obtained from farm interviews, and soil analysis data were used in the calculation of nutrient budgets. Results from modelling indicate declining soil fertility under rainfed agriculture, forest and rangelands, and marginal conditions under irrigated agriculture subject to intensive cultivation. Nutrient deficits were relatively low for irrigated rice-wheat systems, which benefit from nutrient inputs via sediments and irrigation waters, but the introduction of triple cropping showed greater deficits. Nutrient balances were most critical under rainfed maize production where 94% of the farms were in deficit. Current shortages of organic matter make elimination of nutrient deficits problematic but improvement of composting, biological N-fixation and fertilizer efficiency and reducing erosion were found to be potential options.     

beta-glucosidases from five black Aspergillus species: study of their physico-chemical and biocatalytic properties

Five black Aspergillus strains (A. aculeatus, A. foetidus, A. japonicus, A. niger, and A. tubingensis) were cultivated on crude wheat arabinoxylan as the carbon source under defined pH, temperature, and oxygen conditions. Protein and beta-glucosidase content differed remarkably within the obtained culture filtrates, of which eleven beta-glucosidases were isolated. Seven beta-glucosidases were purified to apparent electrophoretic homogeneity using anion-exchange and gel-permeation chromatography. They were found to be acidic proteins and most of them appeared to be glycoproteins with a molecular mass between 93 and 142 kDa. Classification of the beta-glucosidases into four groups (I-A, I-B, II, and III) is suggested according to their physicochemical and biocatalytic properties. The major beta-glucosidases were assigned to groups I-A and I-B, the minor beta-glucosidases to groups II and III, comprising acid-tolerant and glucose-tolerant enzymes, respectively.     

Thymine dimerization in DNA is an ultrafast photoreaction

Femtosecond time-resolved infrared spectroscopy was used to study the formation of cyclobutane dimers in the all-thymine oligodeoxynucleotide (dT)18 by ultraviolet light at 272 nanometers. The appearance of marker bands in the time-resolved spectra indicates that the dimers are fully formed approximately 1 picosecond after ultraviolet excitation. The ultrafast appearance of this mutagenic photolesion points to an excited-state reaction that is approximately barrierless for bases that are properly oriented at the instant of light absorption. The low quantum yield of this photoreaction is proposed to result from infrequent conformational states in the unexcited polymer, revealing a strong link between conformation before light absorption and photodamage.     

Tequila authenticity assessment by headspace SPME-HRGC-IRMS analysis of 13C/12C and 18O/16O ratios of ethanol

By use of headspace SPME sampling and a PLOT column, on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and the pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta(13)C(VPDB) and delta(18)O(VSMOW) values of ethanol in authentic (n = 14) and commercial tequila samples (n = 15) as well as a number of other spirits (n = 23). Whereas with delta(13)C(VPDB) values ranging from -12.1 to -13.2 per thousand and from -12.5 to -14.8 per thousand similar variations were found for 100% agave and mixed tequilas, respectively, the delta(18)O(VSMOW) data differed slightly within these categories: ranges from +22.1 to +22.8 per thousand and +20.8 to +21.7 per thousand were determined for both the authentic 100% agave and mixed products, respectively. The data recorded for commercial tequilas were less homogeneous; delta(13)C(VPDB) data from -10.6 to -13.9 per thousand and delta(18)O(VSMOW) values from +15.5 to +22.7 per thousand were determined in tequilas of both categories. Owing to overlapping data, attempts to differentiate between white, rested, and aged tequilas within each of the two categories failed. In addition, discrimination of tequila samples from other spirits by means of delta(13)C(VPDB) and delta(18)O(VSMOW) data of ethanol was restricted to the products originating from C(3) as well as C(4)/CAM raw materials.     

Clinical and serologic studies of canine borreliosis

During 1984 and 1985, blood samples were obtained from 271 dogs that were suspected of having borreliosis. The dogs lived in areas known to be infested with ticks and had been examined because of limb/joint disorders or for unknown illnesses marked by fever, anorexia, or fatigue. Lameness had been the most frequently reported clinical manifestation. Analyses of serum specimens, by an indirect fluorescent antibody (IFA) method or by an ELISA, detected antibodies to Borrelia burgdorferi, the etiologic agent of borreliosis in dogs and of Lyme disease in human beings. Antibody to B burgdorferi was detected in 76.3% of 114 specimens from dogs living in the lower Hudson Valley region of New York State (predominantly Westchester County), in 66.5% of 155 specimens from dogs from southern Connecticut, and in single specimens from dogs from Rhode Island and California. Geometric mean antibody titers peaked during the winter. Results of IFA tests and ELISA were in agreement, but the latter method yielded less variable results, had greater sensitivity, and was more easily standardized. Five dogs from New York State and Connecticut seropositive to B burgdorferi had developed kidney disorders during or after episodes of intermittent lameness. Application of murine monoclonal antibody in an IFA procedure verified the presence of B burgdorferi in renal cortical tissues from one dog.     

Hunting behavior of a marine mammal beneath the antarctic fast Ice

The hunting behavior of a marine mammal was studied beneath the Antarctic fast ice with an animal-borne video system and data recorder. Weddell seals stalked large Antarctic cod and the smaller subice fish Pagothenia borchgrevinki, often with the under-ice surface for backlighting, which implies that vision is important for hunting. They approached to within centimeters of cod without startling the fish. Seals flushed P. borchgrevinki by blowing air into subice crevices or pursued them into the platelet ice. These observations highlight the broad range of insights that are possible with simultaneous recordings of video, audio, three-dimensional dive paths, and locomotor effort.     

Pulmonary Maturation in Canine Foetuses From Early Pregnancy to Parturition

Development of the foetal respiratory system includes both pulmonary growth and maturation. In human medicine, a higher incidence of respiratory distress is reported in newborn males. This study aimed to identify different phases of canine foetal lung maturation throughout pregnancy, to determine the stage of pregnancy in which surfactant production begins and to compare pulmonary development of male and female foetuses. Pregnant bitches (34) were subjected to elective ovariohysterectomy and allocated into four groups, according to the stage of pregnancy: 30–40 days of pregnancy (n = 10), 41–50 days (n = 10), 51–60 days (n = 10) and bitches in the first stage of parturition (n = 4). Foetal lungs were histologically processed and evaluated by optical microscopy. The pseudoglandular phase was identified between the 35th day and 46th day of gestation; the onset of canalicular and saccular periods was observed, respectively, from the 48th day and 60th day of pregnancy. Lungs from foetuses at term were in the saccular phase; thus, the development into the alveolar period occurs in the neonatal period. The histological analyses revealed that respiratory tract development is centrifugal, from upper to lower airways. Therefore, it is possible to identify distinct development periods in different portions of the same organ. In conclusion, the saccular phase of lung development begins around 57 and 60 days of pregnancy, the period in which surfactant production is believed to occur. Male and female foetuses present similar pulmonary development from early pregnancy until parturition.     

Importance of Staphylococcus hyicus ssp hyicus as a cause of arthritis in pigs up to 12 weeks of age

Lame pigs, up to 12 weeks of age, were necropsied to establish a diagnosis. Of 175 pigs examined, 165 were confirmed to have arthritis by histopathological examination of joint tissues. Lesions were most common in the elbow and tarsal joints and least common in the joints of the feet. Typically, there was severe fibrinopurulent inflammation of synovial membranes regardless of the bacteria isolated. A bacterial aetiology was found in 114 (69%) of the 165 pigs. In arthritic pigs in which an aetiology was established the causative agents were Staphylococcus hyicus ssp hyicus (24.6%), Streptococcus equisimilis (26.3%), Actinomyces pyogenes (13.2%), Staphylococcus aureus (7.9%) and Haemophilus parasuis (7.9%). While gender did not affect the prevalence of arthritis, there was an age influence, most of the pigs culled for arthritis being under 6 weeks of age.