A statistical approach to evaluate compost genotoxicity

Parallel experiments were carried out to evaluate the induction of genotoxic effects in Vicia faba seedlings treated both with aqueous and organic extracts (dimethyl sulphoxide 5%) of 13 composts, or grown on soil samples to which different doses of the same composts were added. Pearsons linear correlation coefficient and multiple regression were used to study how these two different methodological approaches (treatment with the extracts versus direct contact with the composts in soil) agree. In addition, multiple regression analysis was used to investigate the role of heavy metals and humic substances in genotoxic effects and to construct a model to be used to accurately predict potential genotoxicity of the composts. Experimental data obtained at the highest application rate of compost on soil were found to fit the model and evidenced the role of the humic substances in modulating the potential genotoxicity of the different composts.     

WHOLE BODY COMPUTED TOMOGRAPHIC CHARACTERISTICS OF SKELETAL AND CARDIAC MUSCULAR METASTATIC NEOPLASIA IN DOGS AND CATS

Muscular metastatic neoplasia has been reported to be rare in domestic animals, however previous studies were based primarily on necropsy findings. The purpose of this retrospective study was to describe whole body computed tomography (CT) characteristics of confirmed muscular metastases in a cohort of dogs and cats presented for oncology evaluation. Medical records of 1201 oncology patients were reviewed. Included animals underwent pre and postcontrast whole body CT, and CT‐guided tru‐cut biopsy or fine needle aspiration of one or more metastatic lesions. Twenty‐one dogs and six cats met inclusion criteria, representing 2.08% of all canine oncology patients and 3.1% of all feline oncology patients. Mean age was 9.6 years. Postcontrast CT characteristics included well‐demarcated, oval‐to‐round lesions with varying enhancement patterns: ring enhancing (n = 16), heterogeneously enhancing (n = 8), or homogeneously enhancing (n = 5). Five animals showed concurrent and varying nodular patterns. In seven cases (five dogs and two cats), one single muscular nodule was observed. In 20 cases, two or more lesions were observed. In two cases, cardiac hypodense nodules were observed in the postcontrast CT, while appearing isodense in the precontrast study. Necropsy confirmed neoplasia in both of them. Locations of muscular metastases included epaxial/paraspinal muscles of the cervical, thoracic, and lumbar spine (n = 18), superficial muscles of the thoracic wall (n = 13), scapular/shoulder region (n = 3), hind limb (n = 3), and abdominal wall muscles (n = 1). Findings supported the use of pre and postcontrast whole body CT for oncologic staging in dogs and cats, especially for primary tumors characterized by a high metastatic rate.     

Bioaccumulation potential of dietary arsenic, cadmium, lead, mercury, and selenium in organs and tissues of rainbow trout (Oncorhyncus mykiss) as a function of fish growth

The distribution and potential bioaccumulation of dietary arsenic, cadmium, lead, mercury, and selenium in organs and tissues of rainbow trout (Oncorhyncus mykiss Walbaum, 1792), a major aquaculture species, was studied in relation to fish growth over a period of >3 years. Fish were reared under normal farming conditions, that is, fed a standard fish food and exposed to negligible levels of waterborne trace elements. The age-related variations in the content of each trace element in gills, kidney, liver, muscle, and skin were studied through nonparametric regression analysis. A buildup of all elements in all tissues and organs was observed, but due to dilution with growth, the concentrations did not increase, except in a few cases such as cadmium and mercury in liver and kidney. In muscle tissue, the concentrations of mercury, lead, and selenium did not alter significantly with growth, whereas cadmium increased but remained at exceedingly low levels. The concentration of arsenic in muscle tissue peaked at 14 months and then decreased in adult specimens. Arsenic speciation by high-performance liquid chromatography--inductively coupled plasma mass spectrometry revealed that arsenic in muscle was almost exclusively present in the form of nontoxic arsenobetaine. Application of a mercury mass balance model gave predicted concentrations in agreement with measured ones and showed that in farmed rainbow trout the ratio of mercury concentrations in feed and in fish is about 1:1. Therefore, rainbow trout does not approach the limits established for human consumption even when reared with feed at the maximum permitted levels. These findings highlight the low bioaccumulation potential of toxic trace elements such as cadmium, lead, and mercury in rainbow trout following dietary exposure. On the other hand, selenium concentrations in muscle (about 0.2 microg g (-1) of fresh weight) show that rainbow trout may be a good source of this essential element.     

Deactivation of ferrylmyoglobin by vanillin as affected by vanillin binding to β-lactoglobulin

Vanillin was found to be efficient as a deactivator of ferrylmyoglobin with a second-order rate constant of k(2) = 57 ± 1 L mol(-1) s(-1) for reduction to metmyoglobin with ΔH(?) = 58.3 ± 0.3 kJ mol(-1) and ΔS(?) = -14 ± 1 J mol(-1) K(-1) in aqueous pH 7.4 solution at 25 °C. Binding to β-lactoglobulin (βLG) was found to affect the reactivity of vanillin at 25 °C only slightly to k(2) = 48 ± 2 L mol(-1) s(-1) (ΔH(?) = 68.4 ± 0.4 kJ mol(-1) and ΔS(?) = 17 ± 1 J mol(-1) K(-1)) for deactivation of ferrylmyoglobin. Binding of vanillin to βLG was found to have a binding stoichiometry vanillin/βLG > 10 with K(A) = 6 × 10(2) L mol(-1) and an apparent total ΔH° of approximately -38 kJ mol(-1) and ΔS° = -55.4 ± 4 J mol(-1) K(-1) at 25 °C and ΔC(p, obs) = -1.02 kJ mol(-1) K(-1) indicative of increasing ordering in the complex, as determined by isothermal titration microcalorimetry. From tryptophan fluorescence quenching for βLG by vanillin, approximately one vanillin was found to bind to each βLG far stronger with K(A) = 5 × 10(4) L mol(-1) and a ΔH° = -10.2 kJ mol(-1) and ΔS° = 55 J mol(-1) K(-1) at 25 °C. The kinetic entropy/enthalpy compensation effect seen for vanillin reactivity by binding to βLG is concluded to relate to the weakly bound vanillin oriented through hydrogen bonds on the βLG surface with the phenolic group pointing toward the solvent, in effect making both ΔH(?) and ΔS(?) more positive. The more strongly bound vanillin capable of tryptophan quenching in the βLG calyx seems less or nonreactive.     

Chemical composition of the essential oil and supercritical CO2 extract of Commiphora myrrha (Nees) Engl. and of Acorus calamus L

Volatile concentrates from the oleo-gum resin of Commiphora myrrha (Nees) Engl. and from the rhizomes of Acorus calamus were isolated by supercritical extraction with carbon dioxide. The volatile oil of myrrh was obtained at 9.0 MPa and 50 degrees C and at a CO2 flow of 1.5 kg/h. Acorus calamus was extracted at 9.0 MPa and 45 degrees C and at a CO2 flow of 1.6 kg/h. In both cases, an oil devoid of cuticular waxes was obtained with a single depressurization stage. The SFE myrrh oil had a yield, Y, of 3.2%. Its main components, identified and quantified by GC/MS, were furanoeudesma-1,3-diene, 34.9%; lindestrene, 12.9%; curzerene, 8.5%; and germacrone, 5.8%. The essential oils from the same starting material by hydrodistillation, HD, (Y = 2.8%) and by steam distillation, SD, (Y = 0.4%) were quite similar to the SFE extract. The main components of the SFE oil of A. calamus (Y = 3.5%) were acorenone, 13.4%; iso-acorone, 11.6%; (Z)-sesquilavandulol, 11.0%; dehydroxy isocalamendiol, 7.7%; and beta-asarone, 5.5%. The comparison with hydrodistilled (Y = 1.8%) and steam distilled (Y = 1.0%) oils revealed large differences in the content of iso-acorone and crypto-acorone.     

Degradation of alachlor in natural and sludge-amended soils, studied by gas and liquid chromatography coupled to mass spectrometry (GC-MS and HPLC-MS)

Alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl)acetamide] is an herbicide used worldwide. The relative rates of disappearance of alachlor, the formation kinetics of alachlor ethane sulfonic acid (ESA), and the formation of other degradation products in two different soils (a soil with natural organic matter and a sludge-amended soil) has been studied. For such a purpose, soil samples were spiked with alachlor at 2.5 mg kg(-1), concentration generally applied in agricultural soils, and were submitted to sunlight, simulating natural field conditions. Extracts were analyzed by GC-MS and HPLC-MS in scan mode. A good correlation was observed between both techniques, and HPLC-MS allowed the determination of two eluting peaks corresponding to the two stereoisomeric forms of alachlor ESA. Degradation of alachlor in the two soils followed first-order kinetics. Half-life in the natural soil was 4.2 +/- 0.1 days, and half-life in the sludge-amended soil was 5.8 +/- 0.8 days. The higher half-life observed in the sludge-amended soil was attributed to the higher sorption of alachlor to this soil compared to the natural soil. The degradation of alachlor in both soils gave rise to the production of alachlor ESA. Its concentration increased during the incubation period, and after 27 days, its concentration was about 0.59 mg kg(-1) in the natural soil and 0.37 mg kg(-1) in the sludge-amended soil. The other two alachlor transformation products were identified using GC-MS, and the abundance of these degradation products increased while alachlor was degraded.     

Kinetic Approach to Nixtamalization of Corn Pericarp

The nixtamalization of the pericarp isolated from grains of corn was studied with 2, 5, 10, 20 and 40% Ca(OH)₂ based upon the mass of the pericarp. For 2 and 5% Ca(OH)₂ a small loss of the mass of the pericarp takes place quickly which is accompanied by an abrupt fall of the concentration of the OH^‐ ions. A suitable kinetic model for the decrease of the mass of the pericarp is a two phases exponential equation. The second phase is the slowest and it depends on the quantity of Ca(OH)₂ employed.     

Ultrastructural and biomolecular detection of Rickettsiales‐like organisms in tissues of rainbow trout with Red Mark Syndrome

Red mark syndrome (RMS) and US strawberry disease (US SD) are skin disorders affecting rainbow trout farmed in Europe and USA. The disease etiology has not yet been established. In spite of specific investigations, identifying Rickettsia‐like organism (RLO)‐ and Midichloria‐like organism (MLO)‐related DNA in affected individuals, these pathogens have never been observed. We performed histological, ultrastructural and biomolecular analysis on skin and spleen samples of trout with RMS. Examination by TEM revealed the presence of intracytoplasmic microorganisms resembling Rickettsiales within macrophages, fibroblasts and erythrocytes. The microorganisms were oval or short rod shaped (400–800 nm in length and 100–200 nm in width) and often showed a cell wall similar to Gram‐negative bacteria. PCR analysis for Rickettsiales supported these findings: 53% of affected trout were positive by both PCR and TEM The primers RiFCfw‐RiFCrev were used to anneal both the RLO 16S DNA sequence and the MLO 16S DNA sequence. For this reason, and in agreement with previous studies confirming the presence of Rickettsiales‐related DNA in trout with RMS, we assume that TEM detected microorganisms morphologically consistent with bacteria belonging to Rickettsiales order and could be considered as possible causative agents of RMS.     

Early life history of the black anglerfish Lophius budegassa Spinola, 1807 in the Mediterranean Sea using otolith microstructure

The early life history of the black anglerfish, Lophius budegassa was investigated by otolith (lapilli) increment analysis. Samples of demersal juvenile L. budegassa ranging from 54 to 196 mm total length were collected during bottom trawl surveys in the central Adriatic Sea. By counting increments presumed to be deposited daily in the lapillar otoliths, 88 specimens of L. budegassa were successfully aged. Age estimates of juveniles ranged between 79 and 204 days, indicating that probably the pelagic phase of this species is relatively short and settlement occurs at less than 3 months of life. The analysis of check marks in the core area of lapilli enabled us to determine the period of endogenous feeding, which would last between 15 and 24 days after hatching. Back-calculated hatching dates and, consequently, the spawning season of L. budegassa in the Adriatic Sea was spread over a long period, lasting at least from February to June. The length at age relationship gave an estimate of mean growth rate of approximately 0.8–1.02 mm/day, indicating a faster growth rate of 0+ juveniles L. budegassa than previously thought. The implications of these findings on age estimates discrepancies between previous ageing studies on L. budegassa carried out using different calcified structure (sagittae or illicia) are discussed.