Determination of cadmium, copper, and lead in sodium chloride food salts by flame atomic absorption spectroscopy

A method for determination of Cd, Cu, and Pb in sodium chloride food salt samples has been developed. It consists of extraction in 4-methyl-2-pentanone of the complexes formed with ammonium pyrrolidine dithiocarbamate and further analysis of the extracts by flame atomic absorption spectroscopy. Detection limits in ng/g salt were 0.2 for Cd, 0.7 for Cu, and 10.0 for Pb. The coefficients of variation of 12 independent analyses were 13% for Cd (at a level of 0.4 ppb), 18% for Cu (1.6 ppb), and 5% for Pb (40 ppb). The recoveries were 100 +/- 0% for Cd, 115 +/- 14% for Cu, and 100 +/- 13% for Pb.     

Derivative potentiometric stripping analysis (dPSA) used for the determination of cadmium,copper, lead,and zinc in Sicilian olive oils

Derivative potentiometric stripping analysis (dPSA) was utilized to evaluate the Cd(II), Cu(II), Pb(II), and Zn(II) content in olive oil samples produced in Sicily in the crop year 2000-2001. The repeatability of the method was attested at 86.36% for cadmium, at 94.94% for copper, at 99.00% for lead, and at 98.92% for zinc. Recovery tests were carried out, both on cleanup procedures and on extraction steps, on olive oil spiked at different levels; obtained recoveries were 84.52 +/- 9.86 for cadmium, 97.34 +/- 2.72 for copper, 100.68 +/- 0.67 for lead and 83.35 +/- 1.72 for zinc. Theoretic detection limits were 1.2 ng g(-1) for Cd, 3.6 ng g(-1) for Cu, 5.9 ng g(-1) for Pb, and 14.3 ng g(-1) for Zn. Found concentrations range were 15.94-58.51 ng g(-1) for Cu, 32.64-156.48 ng g(-1) for Pb, and 157.00-385.22 ng g(-1) for Zn. Copper, lead, and zinc were found in all samples. The main advantage of this determination consists of a not too aggressive metals extraction procedure using hydrochloric acid, which avoids losses of elements typical of sample calcinations methods.     

Influences of diet and vaccination on colonisation of pigs by the intestinal spirochaete Brachyspira (Serpulina) pilosicoli

The purpose of this study was to determine whether methods used to control swine dysentery (SD), caused by the intestinal spirochaete Brachyspira (Serpulina) hyodysenteriae, would also be effective in controlling porcine intestinal spirochaetosis (PIS) caused by the related spirochaete Brachyspira (Serpulina) pilosicoli. Weaner pigs in Groups I (n=8) and II (n=6) received a standard weaner pig diet based on wheat and lupins, whilst Group III (n=6) received an experimental diet based on cooked white rice and animal protein. Pigs in Group II were vaccinated intramuscularly twice at a 3-week-interval with a formalinised bacterin made from B. pilosicoli porcine strain 95/1000 resuspended in Freund's incomplete adjuvant. Eleven days later pigs in all groups were infected orally with 10(10) cells of strain 95/1000 on three successive days. One control pig in Group I developed acute diarrhoea, and at post-mortem had a severe erosive colitis with end-on attachment of spirochaetes to the colonic epithelium. All other pigs developed transient mild diarrhoea and had moderate patchy colitis at post-mortem 3 weeks later. B. pilosicoli was isolated from the faeces of all pigs, except for one fed rice, and was isolated from the mesenteric nodes of three pigs from Group I and from one vaccinated pig in Group II. Consumption of the rice-based diet, but not vaccination, delayed and significantly (p<0.001) reduced the onset of faecal excretion of B. pilosicoli after experimental challenge. Vaccination induced a primary and secondary serological response to B. pilosicoli, as measured using sonicated whole cells of strain 95/1000 as an ELISA plate coating antigen. Antibody titres in the vaccinated pigs then declined, despite intestinal colonisation by B. pilosicoli. Both groups of unvaccinated animals also failed to develop a post-infection increase in circulating antibody titres.     

Anaplasma infection in free-ranging Iberian red deer in the region of Castilla-La Mancha, Spain

Organisms in the genus Anaplasma are obligate intracellular pathogens that multiply in both vertebrate and invertebrate hosts. The type species, Anaplasma marginale, causes bovine anaplasmosis and infects erythrocytes of the vertebrate host and undergoes a complex developmental cycle in ticks which serve as biological vectors. Infected cattle, wild ruminants and ticks can all serve as reservoirs of A. marginale. In this study, hunter killed Iberian red deer (Cervus elaphus hispanicus) from the region of Castilla-La Mancha in southwestern Spain were tested for Anaplasma infection. We found that 10% of the deer examined were seropositive for Anaplasma. Three A. marginale strains were subsequently obtained from salivary glands of Hyalomma marginatum that were removed from these deer, and the sequence of the major surface protein (msp)4 gene was determined for each strain and used for phylogenetic studies. Maximum parsimony analyses of msp4 sequences from H. marginatum ticks in comparison with New World cattle and bison isolates reported previously, suggested different origins for these Spanish A. marginale strains. The results of this study demonstrated that Iberian red deer are naturally infected with Anaplasma, and may therefore serve as a wildlife reservoir of the pathogen. Although the link between deer infection and the strains of A. marginale identified in ticks was not established, H. marginatum and Rhipicephalus bursa were identified as potential biological vectors for A. marginale in this region and may effect transmission of A. marginale between deer and cattle populations.     

Spatial and temporal patterns at small scale in Austrocedrus chilensis diseased forests and their effect on disease progression

Austrocedrus chilensis is an endemic conifer of Patagonia that suffers a widespread mortality whose causes are a topic of discussion. Since Phytophthora austrocedrae is the most probable cause, we proposed that the spatial and temporal patterns of disease at small scale should reflect pathogen behavior. We aimed at characterizing the spatial and temporal patterns of diseased trees in different soil types and the effect of microsite variability on diseased trees spatial pattern. The spatial pattern of disease was influenced by soil type and tree density. In clay soils with low disease incidence (ca. 25%), the spatial pattern was random and not influenced by abiotic microsite conditions. When disease incidence increased (ca. 70%), concurring with denser plots, the spatial pattern was clustered, as a result of an infection process, and it was independent of microsite variability. In soils with better drainage conditions, that is, alluvial soils with volcanic ash input and coarse textured volcanic soils, the disease was clustered and associated with flat microtopographies. The progression of the disease at small scale was influenced by soil, precipitation and tree density. The spatial and temporal patterns of disease progression were associated with a contagion process and with environmental variables that affect drainage, coinciding with Phytophthora biology and requirements. Our results concur in pointing at Phytophthora as the cause of A. chilensis disease in the study area. Management practices should be urgently applied in order to minimize the spread of the inoculum.     

Inheritance of coffee leaf rust resistance and identification of AFLP markers linked to the resistance gene

The most important disease of Coffea arabica is coffee leaf rust caused by the fungus Hemileia vastatrix. The purpose of this study was to characterize the inheritance of coffee resistance gene(s) to race II of this pathogen and to identify and map molecular markers linked to this trait. Different populations were used: F₂ (160 plants), BCr (20), and BCs (135), derived from a cross between the resistant genotype Híbrido de Timor UFV 427-15 and the susceptible cultivar Catuaí Amarelo UFV 2143-236 (IAC 30). The segregation analysis showed that the resistance of Híbrido de Timor to race II of the H. vastatrix is conferred by a single dominant gene. The amplification of 176 AFLP (Amplified fragment length polymorphism) primer combinations using bulked segregant analysis (BSA) allowed the identification of three molecular markers linked to the resistance gene. Genetic mapping of these three markers in the F₂ population indicated that they are distributed on both sides, flanking the resistance gene. The markers E.CTC/M.TTT405 and E.CGT/M.TGT300 were found linked to the resistance gene at 8.69 cM (LOD 18.91) and 25.10 cM (LOD 5.37), respectively, while E.CCT/M.TTC230 was localized on the other side of the gene, at 20.50 cM (LOD 6.15). These markers are the first rust resistance markers identified in Híbrido de Timor and can be useful for marker assisted selection in coffee breeding programs.