DIAGNOSTIC SENSITIVITY OF ULTRASOUND,RADIOGRAPHY AND COMPUTED TOMOGRAPHY FOR GENDER DETERMINATION IN FOUR SPECIES OF LIZARDS

Gender determination is frequently requested by reptile breeders, especially for species with poor or absent sexual dimorphism. The aims of the current study were to describe techniques and diagnostic sensitivities of ultrasound, radiography, and computed tomography for gender determination (identification of hemipenes) in four species of lizards. Nineteen lizards of known sex, belonging to four different species (Pogona vitticeps, Uromastyx aegyptia, Tiliqua scincoides, Gerrhosaurus major) were prospectively enrolled. With informed owner consent, ultrasound, noncontrast CT, contrast radiography, and contrast CT (with contrast medium administered into the cloaca) were performed in conscious animals. Imaging studies were reviewed by three different operators, each unaware of the gender of the animals and of the results of the other techniques. The lizard was classified as a male when hemipenes were identified. Nineteen lizards were included in the study, 10 females and nine males. The hemipenes were seen on ultrasound in only two male lizards, and appeared as oval hypoechoic structures. Radiographically, hemipenes filled with contrast medium appeared as spindle‐shaped opacities. Noncontrast CT identified hemipenes in only two lizards, and these appeared as spindle‐shaped kinked structures with hyperattenuating content consistent with smegma. Hemipenes were correctly identified in all nine males using contrast CT (accuracy of 100%). Accuracy of contrast radiography was excellent (94.7%). Accuracy of ultrasound and of noncontrast CT was poor (64.3% and 63.1%, respectively). Findings from the current study supported the use of contrast CT or contrast radiography for gender determination in lizards.     

Irrigation effects on quality, phenolic composition, and selected volatiles of virgin olive oils cv. Leccino

Field-grown olive trees (Olea europaea L. cv. Leccino) were used over two growing seasons to determine the effect of deficit irrigation regimes on virgin olive oil (VOO) quality. Drip irrigation was managed to maintain a predawn leaf water potential (PLWP): (a) higher than -1.1 MPa (full irrigation: FI); (b) between -1.0 and -3.3 MPa (deficit irrigation: DI); (c) higher than -4.2 MPa (severe deficit irrigation: SI). The fruit yield and oil yield of DI trees were over 90% of those of FI treatments in both years, respectively, whereas yields of SI trees ranged from 61 to 76%. The irrigation regime had minor effects on the free acidity, peroxide value, and fatty acid composition of VOO. The concentrations of phenols and o-diphenols in VOO were negatively correlated with PLWP. The concentrations of the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol (3,4-DHPEA-EDA), the isomer of the oleuropein aglycon (3,4-DHPEA-EA), and the dialdehydic form of decarboxymethyl elenolic acid linked to (p-hydroxyphenyl)ethanol (p-HPEA-EDA) were lower in FI than in SI treatments. The concentrations of lignans (+)-1-acetoxipinoresinol and (+)-1-pinoresinol were unaffected by the irrigation regime. The tree water status had a marked effect on the concentration of volatile compounds, such as the C(6)-saturated and unsaturated aldehydes, alcohols, and esters.     

Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples

Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health.     

Lipoxygenase distribution in coffee (Coffea arabica L.) berries

In this paper lipoxygenase (LOX) presence was investigated in coffee berries to determine its involvement in lipid degradative metabolism of plants grown in organic and conventional cultivations. An immunochemical analysis has evidenced a ca. 80 kDa protein, cross-reacting with an anti-LOX antibody, only in the pulp fraction of berries obtained from plants of both cultivations. LOX activity in this fraction could be monitored either as conjugated diene formation or reaction products (determined by HPLC) and was mainly associated with a heavy membrane fraction (HMF, enriched in tonoplast, endoplasmic reticulum, plasma membrane, and mitochondria) and a light membrane fraction (LMF, enriched in plasma membrane and endoplasmic reticulum, with low levels of tonoplast and mitochondria). The LOX activity of LMF from berries of both cultivations showed an optimum at pH 8.0. The HMF exhibited a different activity peak in samples from conventional (pH 8.0) and organic (pH 5.5) cultures, suggesting the presence of different isoenzymes. These findings were also confirmed by variation of the ratio of 9- and 13-hydroperoxides in organic (1:1) and conventional cultivations (1:10), indicating that the organic one was subjected to an oxidative stress in the coffee pulp fraction leading to the expression of an acidic LOX. Such de novo synthesized LOX activity could be responsible for the production of secondary metabolites, which may interfere with the organoleptic profile of coffee.     

Supercritical CO2 extract of Cinnamomum zeylanicum: chemical characterization and antityrosinase activity

The volatile oil of the bark of Cinnamomum zeylanicum was extracted by means of supercritical CO2 fluid extraction in different conditions of pressure and temperature. Its chemical composition was characterized by GC-MS analysis. Nineteen compounds, which in the supercritical extract represented >95% of the oil, were identified. (E)-Cinnamaldehyde (77.1%), (E)-beta-caryophyllene (6.0%), alpha-terpineol (4.4%), and eugenol (3.0%) were found to be the major constituents. The SFE oil of cinnamon was screened for its biological activity about the formation of melanin in vitro. The extract showed antityrosinase activity and was able to reduce the formation of insoluble flakes of melanin from tyrosine. The oil also delayed the browning effect in apple homogenate. (E)-Cinnamaldehyde and eugenol were found to be mainly responsible of this inhibition effect.     

Historical and contemporary habitat use of sperm whales around the Galápagos Archipelago: Implications for conservation

1. Sperm whales have occupied the waters off the Galápagos Islands, Ecuador, for at least the past 200 years. During the 19^th century, they were the target of intensive whaling that severely depleted the population. In recent times, after commercial whaling ended, sperm whales in the region remain vulnerable to multiple threats, especially potential entanglement in fishing gear, which may hinder their ability to recover from the whaling era.
2. As a highly mobile, long-lived species, long-term analysis of the habitat use of sperm whales is necessary to establish effective conservation and management strategies. Here, contemporary (1985–2014) and historical (1830–1850) sperm whale habitat use off the Galápagos Islands was analysed and contrasted to the extent of the Galápagos Marine Reserve (GMR). Contemporary habitat use and its variability over time were modelled as a function of geographic, oceanographic, and topographic variables using generalized additive models.
3. The fine-scale habitat (<50 km) used by sperm whales was associated with topographic (i.e. depth and slope) and oceanographic characteristics (i.e. relative sea surface temperature and standard deviation of sea surface temperature), but these preferences varied over time.
4. While historical and contemporary data indicate that sperm whale habitat primarily occurred within the boundaries of the GMR, in recent years, whales were found up to 30.1% of the time outside the GMR, potentially overlapping with commercial fisheries operating in the area.
5. The dynamic nature of the relationship of this nomadic species with its habitat highlights the need of large-scale conservation efforts across the Eastern Tropical Pacific region, including the wide-scale enforcement of regulations requiring the use of Automatic Identification System in fishing vessels, the promotion of on-board fisheries observer programmes, the development of adaptive management strategies, and international collaboration to identify and mitigate threats.

     

Effect of triploidy on quality traits of shi drum (Umbrina cirrosa L.) until the second rearing year

Carcass and flesh morphometric, reological and chemical traits of triploid (3n) and diploid (2n) shi drum (Umbrina cirrosa L.) were evaluated through 7 months. Three age groups, 17‐, 21‐ and 24‐month‐old fish, were investigated. The effects of ploidy were statistically evaluated and the weight of fish was included in the model as a covariate because triploids grew less than diploids. As expected, fish weight was found to be significantly correlated with all the investigated morphometric traits, but showed a negative correlation with some chemical (pH) and colour traits (lightness) of raw fillet. In comparison with controls, triploid shi drums were characterized by different morphological traits that involved a slender body shape. In triploids, a reduction in condition factor, backbone weight, dressing index and an increase in the agility index were also recorded. When the commercial size (i.e. over 300 g) was achieved, triploids exhibited larger coelomatic and fillet (dorsal white muscle) fat deposition than diploids. Among reological traits, colour and texture were affected by ploidy; raw fillet lightness and cooked flesh tenderness were higher among triploids in all the investigated age groups. As fish were fed with a restricted feeding regimen, fillet fat deposition was supposed to be limited. Thus, the effects of ploidy on reological traits may be only partially explained by lipid fillet amount and are supposed to be more related to different fibre muscle architecture.     

Solid olive residues: insight into their phenolic composition

Solid olive residues (SOR) are byproducts of the olive-milling process, but they have an increasing importance in the pharmaceutical industry due to their rich content of biophenols. Such compounds are studied widely for their antioxidant and antimicrobial activities, but there is a lack of information about their quantitative recovery. This research highlighted the key role played both by the selection of the cultivar and by the degree of olive fruit ripening on the phenolic content on the SOR. The extraction methods were selected to reach the best quantitative results mainly using a safe food solvent. In light of the results the Soxhlet extraction with ethanol could be proposed as preferential for a higher recovery of verbascoside and its analogues.     

Bats in a fragmented landscape: Species composition, diversity and habitat interactions in savannas of Santarém, Central Amazonia, Brazil

Species response to forest fragmentation may strongly vary according to ecological requirements, shape, spatial configuration and connectivity of fragments, and the structure of the surrounding matrix. Bats are natural candidates for studies on forest fragmentation. However, the conclusions of such studies are often contradictory. We present the results of a study on the effects of forest fragmentation on bat communities in Alter do Chão, Santarém area, in Central Amazonia, Brazil, an area composed by forests and fragments surrounded by savanna-like vegetation. We compared the species composition between habitats, tested the effect of size, shape and density of trees of forested areas on the presence species and their relative frequency of captures, and investigated the savannas as an ecological barrier for the dispersion of bats. With an effort of 5678 mistnet-hours, we captured 3740 bats (64 species). Multidimensional Scaling indicated no strong separation between the habitats sampled, however, savanna sites were grouped distinctly of fragments and forest sites. Multiple linear regressions indicated no significant correlation between the number of bat species recorded and the size, shape or tree density in forest sites and fragments. There was a significant correlation between the number of captures and the variables tested, explained by the shape of the sites, but not by size or tree density. In general, sites with bigger shape indices had fewer captures. Ten of the 20 most captured species showed no significant difference between the three habitats. The bat communities in Alter do Chão were not strongly affected by forest fragmentation, and the savannas did not appear to act as an ecological barrier to bats.