Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

Uterine Activity after Induction of Hypocalcaemia in the Ovariectomized Camel (Camelus Dromedarius)

Al-Eknah, M.M., Homeida, A.M., Dafalla, E.A., Galil, A.K.A. and Al-Tahir, A.Y., 1997. Uterine activity after induction of hypocalcaemia in the ovariectomized camel (Camelus dromedarius). Veterinary Research Communications, 21 (1), 45-50.Bilateral ovariectomy was performed in three parous, non-pregnant camels. Intrauterine and intra-abdominal pressure changes were recorded using balloon-tipped catheters. Uterine contractions were induced and maintained in the ovariectomized camels by daily intramuscular injections of 5 mg oestradiol benzoate throughout the experimental period. The frequency of uterine contractions varied from 6 to 9 per minute, whereas the amplitude varied from 2 to 3 kPa in all the animals. Inducing hypocalcaemia to a level of 0.5 mmol/L by Na2EDTA reduced the amplitude of the contractions to below 1 kPa (p<0.001). The frequency of the contractions was not affected.     

Screening RAPD primers for molecular taxonomy and cultivar fingerprinting in the genus Actinidia

Summary Eighty ten-base long arbitrary primers were tested for PCR-based DNA amplification of three species of the genus Actinidia (A. deliciosa the kiwifruit, A. chinensis, and A. kolomikta), with the aim of screening species-specific and genotype-specific markers.Of the 80 primers tested, 30 gave an average of 3.5 bands which were monomorphic within one or two species and absent in the remaining one(s), thus resulting in useful markers for taxonomic and phylogenetic purposes. None of the primers tested produced bands linked to sex. Twenty primers out of the twenty-five selected from a preliminary screening showed high levels of polymorphism, producing two to eleven patterns each from the 13 kiwifruit cultivars examined.We found the Stoffel fragment and the Taq polymerase were both suitable for RAPD analysis, the most noticeable difference being the smaller size of fragments (0.4–1.2 kb) produced by the former in comparison to the latter (1.0–3.4 kb). We tested also three different annealing temperatures (35, 37, and 39° C) and found the intermediate one best for number of amplified bands and reproducibility of results.Abbreviations 2-BE 2-butoxyethanol - CTAB hexadecyltrimethylammonium bromide - MAS Marker-Assisted-Selection - PCR Polymerase Chain Reaction - RAPD Random Amplified Polymorphic DNA - RFLPs Restriction Fragment Length Polymorphisms     

Simultaneous determination of Cd(II), Cu(II), Pb(II), and Zn(II) in citrus essential oils by derivative potentiometric stripping analysis

Citrus essential oils are widely used in the food, cosmetics, and pharmaceutical industries, so the determination of heavy metals content is of great importance to guarantee their quality. The present work deals with the quantification of Cd(II), Cu(II), Pb(II), and Zn(II) in different varieties of citrus essential oils, using derivative potentiometric stripping analysis. Two different metals extraction procedures, involving concentrated hydrochloric acid treatment and acid-alcoholic dissolution, are tested on lemon, mandarin, sweet orange, and bergamot essential oils, and they give very similar results. Cd(II), Cu(II), Pb(II), and Zn(II) recovery tests spanned from 95 to 100.50%, providing evidence that metals quantification remained unaffected by the cleanup steps of the two procedures. The repeatability of the hydrochloric acid extraction method, applied on different varieties of essential oils, is >95.00% for Cd(II), Cu(II), Pb(II), and Zn(II), whereas the repeatability of the acid-alcoholic dissolution method is >93.00% for Cu and Cd only in lemon oil. Detection limits obtained for the four analytes, using both procedures, ranged from 0.10 to 0.98 ng g(-)(1) in lemon, mandarin, sweet orange, and bergamot essential oils.     

Determination of some heavy metals and selenium in Sicilian and Calabrian citrus essential oils using derivative stripping chronopotentiometry

This paper aims to bring some novelty about the concentration of some heavy metals and selenium in biological citrus essential oils (CEO) produced in Sicily and Calabria in different crop years. Derivative stripping chronopotentiometry has been used as an accurate, sensitive, and rapid technique for the determination of Cd, Cu, Mn, Ni, Pb, Zn, and Se in hydrochloric acid extracts of CEO; in the optimized electrochemical conditions, detection limits of <1 microg kg(-1) were obtained for all of the studied metals. In particular, the concentrations of metals were determined in biological bergamot essential oils produced in Calabria in 1999 and 2000 and in biological CEO produced in Sicily in 2003 and 2004. The obtained results provided evidence that Mn was the most abundant metal in all of the studied CEO followed by Zn, Ni, Cu, Pb, and Se; Cd concentrations were always lower than the limit of detection (0.6 microg kg(-1)).     

Protective effect of the AT137RQ and ARQK176 PrP alleles against classical scrapie in Sarda breed sheep

The susceptibility of sheep to scrapie is under the control of the host’s prion protein (PrP) gene and is also influenced by the strain of the agent. PrP polymorphisms at codons 136 (A/V), 154 (R/H) and 171 (Q/R/H) are the main determinants of susceptibility/resistance of sheep to classical scrapie. They are combined in four main variants of the wild-type ARQ allele: VRQ, AHQ, ARH and ARR. Breeding programmes have been undertaken on this basis in the European Union and the USA to increase the frequency of the resistant ARR allele in sheep populations. Herein, we report the results of a multi-flock study showing the protective effect of polymorphisms other than those at codons 136, 154 and 171 in Sarda breed sheep. All ARQ/ARQ affected sheep (n = 154) and 378 negative ARQ/ARQ controls from four scrapie outbreaks were submitted to sequencing of the PrP gene. The distribution of variations other than those at the standard three codons, between scrapie cases and negative controls, was statistically different in all flocks. In particular, the AT₁₃₇RQ and ARQK₁₇₆ alleles showed a clear protective effect. This is the first study demonstrating a protective influence of alleles other than ARR under field conditions. If further investigations in other sheep breeds and with other scrapie sources confirm these findings, the availability of various protective alleles in breeding programmes of sheep for scrapie resistance could be useful in breeds with a low frequency of the ARR allele and would allow maintaining a wider variability of the PrP gene.     

Comparison of isothiocyanate metabolite levels and histone deacetylase activity in human subjects consuming broccoli sprouts or broccoli supplement

Increased consumption of cruciferous vegetables such as broccoli may reduce the risk of various cancers. Myrosinase is required to convert dietary glucosinolates from broccoli into bioactive isothiocyanates. We evaluated isothiocyanate excretion profiles in healthy subjects who consumed broccoli sprouts or broccoli supplement (no myrosinase) with equivalent glucosinolate content. Urinary metabolites of two major isothiocyanates, sulforaphane and erucin, were measured by liquid chromatography coupled with tandem mass spectrometry. Peak excretion of sulforaphane and erucin was higher and occurred sooner in subjects who consumed broccoli sprouts as compared to subjects who consumed the supplement. A subject-dependent shift in the ratio of urinary sulforaphane to erucin metabolites was observed in both groups, indicating conversion of sulforaphane to erucin. Lower histone deacetylase activity was observed in the peripheral blood mononuclear cells only in subjects consuming sprouts. Fresh broccoli sprouts differ from broccoli supplements in regards to excretion of isothiocyanates and bioactivity in human subjects.     

Oxygen affinity and Bohr effect responses to 2,3-diphosphoglycerate in equine and human blood

The dependence of blood oxygen affinity and the Bohr effect on the concentration of 2,3-diphosphoglycerate (DPG) in erythrocytes was investigated in 24 trotter horses and 24 healthy men. The oxygen tension at half saturation and standard conditions (P₅₀st at pH 7·4 PCO₂40 mniHg and 37°C) and the carbon dioxide or fixed-acid-induced Bohr effect (dlogP₅₀/dpH) were determined. Samples of fresh blood and blood depleted of or enriched with DPG were studied. In the absence of measurable DPG, the equine and human blood had similar mean (SD) values of P₅₀st (16·6 [0·6] and 16·2 [0·7] mmHg, respectively). In both species these values increased with increasing DPG, but the response of equine blood was significantly lower, at least up to physiological values (P₅₀st =24·6 [0·6] and 26·2 [0·7]) mmHg; DPG=14([1·8] and 12·8 [1·2] μmol gHb⁻¹, respectively, in fresh blood). concentrations above 20 to 25 μmol gHb⁻¹ of DPG the difference between the values of P₅₀st in the two species tended to de crease because the response in human blood reached a plateau. The interactions between the Bohr effect and the concentration of DPG showed that in the horses, as in the men, the level of DPG played an important role in governing the relative magnitude of carbon dioxide and fixed acid factors. The difference between them, which is associated with the oxylabile carbamino binding, was greatest in DPG-depleted blood, but whereas in the men the difference was suppressed by an above normal DPG concentration, in the horses it was still measurable.