Mass spectrometry of intact V-type ATPases reveals bound lipids and the effects of nucleotide binding

The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.     

Detection, accumulation, distribution, and depletion of furaltadone and nifursol residues in poultry muscle, liver, and gizzard

Nitrofurans were broadly used as an extremely effective veterinary antibiotic especially in pig and poultry production farms. Because of fears of the carcinogenic effects on humans, the nitrofurans were banned from use in livestock production in many countries, including the European Union. The present study examines the accumulation, distribution, and depletion of furaltadone and nifursol and of their tissue-bound metabolites [3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) and 3,5-dinitro-salicylic acid hydrazine (DNSAH), respectively, in poultry edible tissues (muscle, liver, and gizzards) following administration to chickens of therapeutic and subtherapeutic concentrations of both compounds. Nitrofurans determination was performed by high-performance liquid chromatography-diode array detection and liquid chromatography-tandem mass spectrometry, respectively, for feeds and for poultry tissues. Furaltadone and nifursol, in very low concentrations, were found in samples of muscle, liver, and chicken's gizzard collected from slaughtered animals after 5 weeks of treatment and no withdrawal time period. When a withdrawal time period of 3 weeks was respected, no detectable nitrofuran parent compounds was observed in all of the studied matrices. For AMOZ, concentrations of 270 μg/kg in meat, 80 μg/kg in liver, and 331 μg/kg in gizzard were determined after administration of a medicated feed with furaltadone (132 mg/kg), 3 weeks after withdrawal of treatment. For DNSAH, the concentration values obtained are much lower than those observed for AMOZ. For meat, liver, and gizzard, DNSAH concentrations of 2.5, 6.4, and 10.3 μg/kg, respectively, were determined, after administration of a medicated feed with nifursol (98 mg/kg), 3 weeks after withdrawal of treatment. The gizzard could be considered a selected matrix for nitrofuran residues evaluation in poultry, due to its capacity of retaining either nitrofuran parent compounds or metabolites in higher concentrations, regardless of the administered dose or of the respected withdrawal time period.     

Long-term effect of toltrazuril on growth performances of dairy heifers and beef calves exposed to natural Eimeria zuernii and Eimeria bovis infections

The long-term effects of a toltrazuril treatment against natural Eimeria bovis and/or Eimeria zuernii infections were investigated in comparison with diclazuril and untreated controls on two dairy (Italian Friesian breed) and two beef (Chianina breed) farms. At each trial site, 30 calves were allocated into three groups of 10 calves each: T (treated with toltrazuril), D (treated with diclazuril) and C (left untreated). For 40 weeks post-treatment, the calves were weighed and examined clinically and parasitologically. The oocyst counts as well as the number of scour days were significantly lower in the T group than in the D and C groups. Final bodyweights and body condition scores of the T group exceeded those of groups C and D. The results confirmed that toltrazuril was highly efficacious, safe and provided productive benefits in dairy and beef calves.     

Measurement of corneal sensitivity in 20 ophthalmologically normal alpacas

Objective To determine corneal sensitivity in 20 healthy adult alpacas (40 eyes) in order to establish reference values. Design Prospective study. Animals Twenty healthy, adult alpacas. Procedures Corneal sensitivity was determined by the corneal touch threshold (CTT) using a Cochet‐Bonnet esthesiometer. Five different regions of the cornea were evaluated (nasal, ventral, lateral, dorsal, and central). Results Corneal touch threshold values (in mm filament length) obtained from five corneal regions demonstrated varying corneal sensitivities. The central region (34.5 ± 7.1 mm) was the most sensitive, followed by the ventral (29.5 ± 7.2 mm), medial (29.3 ± 7.3 mm), dorsal (25.6 ± 6.3 mm), and lateral (21.8 ± 5.7 mm) regions. All pairwise comparisons of regional differences were statistically significant (P < 0.05), except for the comparison between the ventral and medial regions (P = 0.88). Evaluation of the CCT while controlling for age (2–5 years vs. 6–10 years) demonstrated an increased CTT with an increasing age for the central, ventral, medial and lateral regions. A decrease in the CTT of the dorsal region was noted with increasing age. Conclusion and Clinical Relevance Corneal touch threshold values in 40 eyes of 20 healthy adult alpacas were determined using a Cochet‐Bonnet esthesiometer. This study demonstrated the central corneal region to be most sensitive. Values obtained may serve as reference values in subsequent studies.     

Pathogens of bark beetles (Coleoptera: Curculionidae) in Bulgarian forests

The occurrence and prevalence of bark beetle pathogens in forest stands in Bulgaria were investigated in 944 specimens belonging to 21 bark beetle species. Protozoa, microsporidia, fungi and nematodes occurred in 19 of all investigated species. The infections were found in the gut (nematodes, gregarines, microsporidia), gonads (microsporidia) and hemolymph (nematodes) of the infected insects. Protozoan species (Gregarina typographi, Gregarina spp.) were detected in eight bark beetle species. Morphometric data about G. typographi and Gregarina spp. are presented. The prevalence of the gregarines varied between 1.4% and 64.2%. Microsporidia of the genera Nosema and Chytridiopsis were revealed in three bark beetle species. The prevalence of microsporidia ranged between 1.5% and 11.8%. This is the first report of a microsporidium in Taphrorychus villifrons and of gregarines in T. villifrons, Pityogenes bistridentatus, P. conjunctus, and Orthotomicus erosus. The fungus Beauveria bassiana was found in 3.4% of Hylurgops palliatus specimens. Nematodes (in gut and haemolymph) were revealed in 19 bark beetle species and their prevalence varied between 10% and 98.5%.     

Hydrogeochemistry and Water Balance in the Coastal Wetland Area of “Biviere di Gela,” Sicily, Italy

In the study area physical and chemical factors control the composition of surface and groundwaters, which in turn determine the water quality of the “Biviere di Gela” lake. These factors combine to create diverse water types which change their compositional character spatially as rainfall infiltrates the soil zone, moves down a topographically defined flow path, and interacts with bedrock minerals. Low-salinity waters, which represent the initial stage of underground circulation, start dissolving calcium carbonate from the local rocks. The progressive increase in salinity, characterized by substantially higher Ca, SO₄, Na and Cl concentrations, suggests that dissolution of CaSO₄ and NaCl is an important process during water–rock interactions. The “Biviere di Gela” lake is often separated into two units (Lago Grande and Lago Piccolo). “Lago Grande” water is generally of Na-SO₄-Cl-type, whereas “Lago Piccolo” water is of Na-Cl-SO₄-type. Their total content of dissolved salts varies with season, the amount of rainfall, and inflow of ground and drainage water. Over time, an increasing trend towards greater salinity and also sudden changes in the relative abundances of Cl and SO₄ have been recorded for the “Lago Grande.” The isotope composition of the lake water appears to be affected by inflow of ground and surface waters and also by evaporative loss. The nitrate content of waters from the recharge basin is of particular concern because it contributes to lake eutrophication. The trace element contents do not evidence the presence of any significant metal contamination of lake waters, although a future potential hazard of metals bioaccumulation by the aquatic biota must be taken into consideration. Finally, a water balance for the basin shows that a drop in precipitation of about 20% might be critical for lake survival.     

Suicide vectors for the introduction of genetic markers into Bradyrhizobium strains by site-directed chromosomal integration between repeated sequences (RS-α)

To achieve stable expression of the heterologous and reporter genes in the Bradyrhizobium genome, we constructed suicide plasmids capable of site-directed genomic integration of the gusA, gfp and nifA genes by homologous recombination into non-essential repeated sequences (RS-α), isolated from B. japonicum strain CPAC7 (SEMIA5080). In this report, we describe the strategies to construct the vectors and their use to obtain mutants with site-specific insertions.     

Removal of Copper and Cadmium Ions from Diluted Aqueous Solutions by Low Cost and Waste Material Adsorbents

The sorption of copper and cadmium ions using activated carbon,kaolin, bentonite, diatomite and waste materials such as compost,cellulose pulp waste and anaerobic sludge as sorbents is reported. Equilibrium isotherms were obtained for the adsorption of these metals in single and binary solutions. Bentonite presented the highest adsorption capacities for both copper and cadmium. A competitive uptake was observed when both metals are present; copper being preferentially adsorbed by all materials with theexception of anaerobic sludge. Equilibrium data were fitted toLangmuir and Freundlich models, with satisfactory results for most of the adsorbent-metal systems studied.Of all the adsorbents studied, bentonite and compost presentedthe highest removal efficiencies, reaching 99% for copper whencadmium is also present, for initial solution concentrations ofup to 100 mg L^-1. Anaerobic sludge has a greater preferencefor cadmium, even in the presence of copper, with removal efficiencies of 98% for similar concentrations to those mentioned above.     

Linalool from Lippia alba: study of the reproducibility of the essential oil profile and the enantiomeric purity

A new chemotype of the aromatic Verbenaceae species Lippia alba Mill. N. E. Br. from southeastern Brazil has recently been shown to have a high content of linalool in the leaf essential oil. Vegetative propagation of this chemotype was conducted at six different locations in Brazil, and the variation of the content and the optical purity of linalool in the oils were verified. Yields (0.6-0.9%, hydrodistillation), chemical composition, linalool content, and optical purity of the oils from all the plants were compared, using GC-FID, GC-MS, chiral chromatography, and retention index calculation. No plant exceeded the matrix in linalool content (46.5 to 90.7%), and the chemical profile of the oils was the same for all the samples. Purification of linalool to a content close to 100% was effected by vacuum distillation of the crude oil. Chiral analysis showed exclusively the presence of S-linalool in all the crude oils and in the distilled samples.     

URETEROLITHIASIS AND PAPILLOMA FORMATION IN THE URETER OF A GUINEA PIG

A 5-year-old guinea pig with suspected urolithiasis was presented for radiology and ultrasound examinations of the abdomen. Radiographically, an irregular-shaped mineral opacity was detected in the area of the urinary bladder. Ultrasonographically, pyelectasia of the right kidney, hydroureter with an ureterolith cranial to a thickened ureter wall close to the ureterovesical junction, and a thickened urinary bladder wall were detected. Histopathologically, the thickened ureter wall was found to be a papilloma. The ureter calculus consisted of 100% calcite.