Determination of organic and inorganic carbon, δ13C,and nitrogen in soils containing carbonates after acid fumigation with HCl

In carbonate‐containing soils a reliable determination of organic C requires a method that effectively separates organic and inorganic C without altering the organic matter. This study was conducted to determine whether HCl vapor completely removes carbonates even in dolomite‐rich soils and to what extent a widely used acid‐fumigation method has to be modified for humus‐rich soils. Furthermore, it was tested whether HCl fumigation alters organic‐C content. Since C and N parameters are often analyzed simultaneously we also tested the influence of acid‐vapor treatment on N content and on δ¹³C of soil organic matter. We applied fumigation with 37% HCl for 8 and 32 h using 9 carbonate‐containing soil samples. Inorganic C ranged from 7 to 124 and organic C from 9 to 267 g kg^–1. The maximum contents of dolomite and calcite were 940 and 640 g kg^–1, respectively. A time of 8 h was enough to completely remove all carbonates. Neither the content nor the δ¹³C of organic C were significantly affected by fumigation. In contrast, N contents were altered by acid treatment. Based on these results and on our experience in analyzing more than 1000 soil samples, a recommended procedure for acid fumigation of carbonate‐containing soils with a wide range of organic‐ and inorganic‐C contents was derived. Samples pretreated in this way can be analyzed reliably for their organic‐C content and δ¹³C. Furthermore, N and inorganic‐C contents can be determined with a quality sufficient for many purposes.     

Molecular characterization of Mycobacterium tuberculosis complex isolates from wild ungulates in south-central Spain

The role of European wild ungulates in the epidemiology of tuberculosis (TB) is still under discussion. This study describes the geographical distribution and molecular typing of 77 Mycobacterium tuberculosis complex isolates belonging either to M. bovis or to M. caprae, cultivated from hunter harvested red deer (Cervus elaphus) and European wild boar (Sus scrofa) in 24 Spanish localities, and compares them with spoligotypes detected previously in humans, livestock or wild animals, as described in the literature. The distribution of the molecular type patterns suggests that the population of M. tuberculosis complex strains isolated from Spanish wild ungulates is spatially structured despite the lack of important geographical barriers and despite the increasingly frequent wildlife translocations. Red deer and the European wild boar can share the same molecular types in localities in which the M. tuberculosis complex was isolated from both species. Strains of bovine and caprine origin do circulate in the same local wildlife populations. Six out of 11 spoligotypes were similar to types described in human cases. The isolation of TB strains in fenced estates from wild animals that have not had contact with domestic livestock for at least the past two decades, strongly suggests that the M. tuberculosis complex is able to survive in these populations. Therefore, wildlife including cervids and the wild boar need to be considered in the epidemiology and control of tuberculosis.     

Purification and partial characterization of feline pepsinogen

OBJECTIVE: To purify and partially characterize feline pepsinogen (fPG) from the gastric mucosa and compare fPG with PGs of other species. SAMPLE POPULATION: Stomachs of 6 cats. PROCEDURE: A crude protein extract was prepared from the gastric mucosa of feline stomachs. Feline PG A was purified by ammonium sulfate precipitation, weak-anion-exchange chromatography, size-exclusion chromatography, and strong-anion exchange chromatography. Partial characterization consisted of estimation of molecular weights (MWs) and isoelectric points, N-terminal amino acid sequencing, and investigation of susceptibility to pepstatin inhibition. RESULTS: Several fPG A-group isoforms were identified. The MWs of the isoforms ranged from 37,000 to 44,820. Isoelectric points were all < pH 3.0. The proteolytic activity of the activated PGs was inhibited completely by pepstatin in a range of equimolar to 10-fold molar excess. The specific absorbance of fPG A was 1.29. The N-terminal amino acid sequence of the first 25 residues of the predominant fPG A7 had 75%, 72%, 64%, and 56% homology with PG A of dogs, rabbits, cattle, and humans, respectively. Sequences of 4 other fPG A-group isoforms were similar to fPG A7. All isoforms were immunologically cross-reactive with sheep anti-fPG A7 antiserum. CONCLUSIONS AND CLINICAL RELEVANCE: PG A is the only identified type of PG in cats and, similar to pg in other species, comprises multiple isoforms. The availability of fPG A may be used to facilitate the development of an immunoassay to quantify serum fPG A as a potential marker for gastric disorders in cats.     

Wild boar helminths: risks in animal translocations

The helminth populations found in a group of wild boars collected in central Spain were compared to those in a group of animals imported from a French game farm that produces boars for restocking. Eleven helminth species, including ten nematodes and one acanthocephalan, were found. Gongylonema pulchrum and Macracanthorhynchus hirundinaceus were only detected in autochthonous wild boars, while Oesophagostomum dentatum, Ascaris suum, and Trichuris suis were detected in imported animals only. Autochthonous wild boars were more frequently and more intensely parasitised by Ascarops strongylina than the imported ones. No differences in prevalence nor intensity were found for the species Capillaria garfiai, Globocephalus urosubulatus, Metastrongylus sp., Physocephalus sexalatus and Simondsia paradoxa. To our knowledge, G. urosubulatus, G. pulchrum and S. paradoxa have not previously been described in wild boars in Spain. Our results highlight the risks of translocating wild animals, with regard to their helminth parasites. Until improved control measures are established, it would be wise to avoid long-distance translocations in order to prevent the potential introduction of foreign parasites.     

Effects of transcorneal iridal photocoagulation on the canine corneal endothelium using a diode laser

Objective To investigate the potential damage to the canine corneal endothelium following transcorneal iridal laser photocoagulation using a semiconductor diode laser. Animals studied Sixteen young mongrel dogs. Procedures Baseline corneal endothelial cell counts and corneal thickness were measured in the central and temporal quadrants using a noncontact specular microscope under general anesthesia. Transcorneal iridal photocoagulation was applied using a semiconductor diode laser in a continuous mode with the use of an operating microscope. Fifteen dogs were treated, and the sixteenth dog served as a control. Fifteen different treatment combinations were randomly assigned to the 30 eyes; the fellow eye was treated differently. Three treatment factors were investigated: (1) laser energy intensity, (2) target tissue to endothelial distance, and (3) laser application duration. After 3 weeks the dogs were euthanized, specular microscopy was repeated, and the cornea was examined by scanning electron microscopy. Results Dyscoria and focal iris darkening were noted in all eyes immediately following laser treatment. Focal corneal edema (n = 2) and an incipient anterior capsular cataract (n = 1) were also noted. Baseline mean corneal endothelial cell densities were 2530 cells/mm² centrally and 2607 cells/mm² temporally. Postlaser corneal endothelial cell densities were 2499 cells/mm² centrally and 2523 cells/mm² temporally. Mean prelaser corneal thickness measurements were 0.555 mm centrally and 0.549 mm temporally. Postlaser corneal thickness measurements were 0.580 mm centrally and 0.554 mm temporally. Statistical analyzes revealed no significant changes in endothelial cell densities (P > 0.05) or corneal thickness (P > 0.05) induced by any treatment combination. Aside from tissue handling and processing artifacts, scanning electron microscopy revealed no endothelial cell damage. Conclusions Our study demonstrated by specular and scanning electron microscopy that diode laser iridal photocoagulation had no significant effect on the canine corneal endothelium within the parameters described. However, one must take into consideration the young age of the dogs and the potential for corneal endothelial cell regeneration in young dogs, and the relatively short period of postoperative study.     

Nitrogen best management practice for citrus trees: I. Fruit yield,quality, and leaf nutritional status

Elevated levels of nitrate-nitrogen (NO₃-N) in the surficial aquifer above the drinking water quality standard, i.e. maximum contaminant limit (MCL; 10 mg L⁻¹), have been reported in some part of central Florida citrus production regions. Soils in this region are very sandy (sand content >95%), hence are vulnerable to leaching of soluble nutrients and chemicals below the rooting depth of the trees. The objective of this research was to develop N and irrigation best management practices for citrus in sandy soils to maintain optimal crop yield and quality, and to minimize potential leaching of nitrate below the root zone. Six years of field experiment was conducted in a high productive (mean fruit yield > 80 Mg ha⁻¹yr⁻¹) >20-year-old ‘Hamlin’ orange trees [Citrus sinensis (L.) Osbeck] on ‘Cleopatra mandarin’ (Citrus reticulata Blanco) rootstock grown on a well drained Tavares fine sand (hyperthermic, uncoated, Typic Quartzipsamments) in Highland county, FL. Nitrogen rates ranged from 112 to 280 kg ha⁻¹ yr⁻¹ applied as fertigation (FRT), water soluble granular (WSG), 50:50 mix of FRT and WSG, and controlled-release fertilizer (CRF). Tensiometers were used to monitor the soil water content as a basis to schedule optimal irrigation. Fruit yield response over the entire range of N rates was greater for the FRT and WSG sources as compared to that for the WSG + FRT or CRF sources. Using the regression analysis of the fruit yield in relation to N rate, the optimum N rate appeared to be at 260 kg ha⁻¹ yr⁻¹. Based on fruit production response in this study, the N requirement for production of 1 Mg of fruit varied from 2.2 to 2.6 kg across four N sources. This study demonstrated an increased N uptake efficiency, as a result of best management of N and irrigation applications. The optimal N and K concentration in the 4–6-month-old spring flush leaves were 26–30, and 15–18 g kg⁻¹, respectively. However, fruit yield response showed no significant relationship with concentrations of P in the 4–6-month-old spring flush leaves over a range of 0.8–2.4 g kg⁻¹. The results of fate and transport of N in soil and in soil solution with application of different rates and sources of N, and components of citrus tree N budget, are reported in a companion paper.     

Partial purification and properties of an esterase from tomato cell suspension cultures hydrolysing the pyrethroid insecticide cyfluthrin

An esterase which hydrolyses the pyrethroid insecticide cyfluthrin, was isolated from tomato cell suspension cultures and purified 10-fold. The apparent molecular weight of the enzyme was estimated to be 32 000 Dalton, the pH-optimum 8-0, and the temperature optimum 35°C. The esterase showed a low substrate specificity and hydrolysed certain esters, e. g. ethyl 4-nitrobenzoate, p-nitrophenyl acetate and dinoseb acetate, whereas ethyl butyrate and the organophosphates demeton-S-methyl sulfoxide and paraoxon could not be hydrolysed. Because of the additional strong inhibition of the enzyme by these organophosphates the cyfluthrin hydrolase is suggested to belong to the B-esterase type. An apparent K_m-value of 6-25 × 10^?4 mol litre^?1 was obtained for p-nitrophenyl acetate as substrate.     

Comparative histopathology of the chitin synthesis inhibitors nikkomycin X/Z,nikkomycin Z and polyoxin D. I: Effects on moulting,reproduction and digestion in the spider mite Tetranychus urticae

The effects of the closely related chitin synthesis inhibitors, the nucleoside peptidyl antibiotics nikkomycin X/Z, nikkomycin Z and polyoxin D on the two-spotted spider mite, Tetranychus urticae, have been studied by electron microscopy. The application of all three compounds, nikkomycin X/Z (95% active ingredient, ratio of components unknown), nikkomycin Z (pure component) and polyoxin D (pure component) disrupted cuticle, eggshell and yolk synthesis and influenced the silk and salivary glands. Ultrahistological patterns indicated differences in the mode of action of the three compounds which were especially evident in an alteration of the nuclear ultrastructure in the mid-gut cells, which was only caused by polyoxin D. Mechanisms of action and their consequences on function, based on previously published information on the action of these compounds, are discussed.     

Cytoskeletal changes in <Emphasis Type="Italic">Eimeria bovis</Emphasis>-infected host endothelial cells during first merogony

The first merogony of Eimeria bovis takes place in lymphatic endothelial cells of the ileum, resulting in the formation of macromeronts up to 250 mum. In this study, we investigated the host cell cytoskeleton (actin filaments, microtubules, spectrin, vimentin intermediate filaments) associated with parasitic development in vitro by confocal laser scanning microscopy (CLSM) using primary bovine umbilical vein endothelial cells (BUVEC) and bovine spleen lymphatic endothelial cells (BSLEC) as host cells. No prominent changes in the host cell cytoskeleton were detected 1-3 days after E. bovis sporozoite invasion. With ongoing meront maturation a significant increase in microtubules and actin filaments close to the parasitophorous vacuole (PV) was found. Mature macromeronts within the PV were completely enclosed by these cytoskeletal elements. Our findings suggest, that in order to guarantee the survival of the host cell on the enlargement of macromeronts, E. bovis needs not only to augment but also to rearrange its cytoskeletal system.     

Pattern of inheritance of a self-fertility gene in an autotetraploid perennial ryegrass (Lolium perenne) population

A mutation causing self-fertility (SF) in perennial ryegrass was studied at the tetraploid level. The aim of this work was to determine a) whether SF remains functional in a tetraploid population and b) whether the SF mutation expresses dominance in heterozygous pollen grains. A tetraploidized plant carrying SF alleles was self-pollinated to create a segregating F₂ population. In the F₂ individuals, pollen compatibility ranged between 38% and 84% showing that SF remained functional. The SF locus genotype was the main determinant of pollen compatibility explaining 78% of the variation. The observed segregation was significantly different from the expected under both SF being dominant or recessive models (P_(χ2)≤0.001) and tended to be intermediate between them, indicating partial dominance or additive gene action. The frequency of the different genotypes suggested that pollen grains homozygous for the mutation have a competitive advantage over heterozygous pollen and that pollen compatibility is affected by the interaction with additional loci. The implications of our results for breeding polyploid grasses are discussed.