Cervids as sentinel‐species for tick‐borne encephalitis virus in Norway ‐ A serological study

Tick‐borne encephalitis virus (TBEV) is the causative agent of tick‐borne encephalitis (TBE). TBEV is one of the most important neurological pathogens transmitted by tick bites in Europe. The objectives of this study were to investigate the seroprevalence of TBE antibodies in cervids in Norway and the possible emergence of new foci, and furthermore to evaluate if cervids can function as sentinel animals for the distribution of TBEV in the country. Serum samples from 286 moose, 148 roe deer, 140 red deer and 83 reindeer from all over Norway were collected and screened for TBE immunoglobulin G (IgG) antibodies with a modified commercial enzyme‐linked immunosorbent assay (ELISA) and confirmed by TBEV serum neutralisation test (SNT). The overall seroprevalence against the TBEV complex in the cervid specimens from Norway was 4.6%. The highest number of seropositive cervids was found in south‐eastern Norway, but seropositive cervids were also detected in southern‐ and central Norway. Antibodies against TBEV detected by SNT were present in 9.4% of the moose samples, 1.4% in red deer, 0.7% in roe deer, and nil in reindeer. The majority of the positive samples in our study originated from areas where human cases of TBE have been reported in Norway. The study is the first comprehensive screening of cervid species in Norway for antibodies to TBEV, and shows that cervids are useful sentinel animals to indicate TBEV occurrence, as supplement to studies in ticks. Furthermore, the results indicate that TBEV might be spreading northwards in Norway. This information may be of relevance for public health considerations and supports previous findings of TBEV in ticks in Norway.     

Endometrial angiogenesis induced by uterine insufflation with an oxygen–ozone gas mixture in mares

Ozone (O₃) therapy has been used to improve peripheral tissue oxygenation in humans and domestic animals. The goal of the present study was to characterize histological changes in the endometria of healthy equines following tissue exposure to gas mixtures enriched with different concentrations of O₃. Cycling mares without endometrial degeneration were divided into three groups according to treatment (n = 9 mares/group). The uteri from the O₃, ½O₃ and control groups were insufflated for 3 min with gas containing 42, 21 and 0 μg O₃ ml^-1, respectively. Treatments were performed every three days from D0 to D6. Endometrial samples were collected immediately before the first treatment and 24 hr after the last treatment. The following nine histological parameters were evaluated: (i) the number of endometrial blood vessels, (ii) endometrial vascular degree (EVD), (iii) increase rate of blood vessels, (iv) increase rate of EVD, (v) glandular total area, (vi) glandular lumen area, (vii) intraglandular secretion area, (viii) glandular epithelial height and (ix) luminal epithelial height. In the O₃ group, a positive effect from treatment (p < .01) was detected for all vascular parameters (i, ii, iii and iv), glandular total area, intraglandular secretion area and glandular epithelial height. Compared to the control group, the ½O₃ group had greater (p < .01) EVD (84.1 ± 12%) and a higher increase rate of blood vessels (151.9 ± 47.1%). Uterine insufflation with low or intermediate concentrations of the O₂–O₃ gas mixture induced endometrial angiogenesis. Morphometry, but not morphology, of the endometrial glands was affected by local O₃ therapy. These findings would be of great significance for the development of new therapies for infertility in mares.     

Using the scaling behaviour of higher taxa for the assessment of species richness

In this paper, we show that fine-scale data on seed plant diversity exhibit exponential scaling behaviour across the taxonomic hierarchy. One practical implication of this result is that the scaling behaviour of higher taxonomic levels can be used to predict species richness in a given area with reasonable accuracy.     

Comparison of yield, composition, and antioxidant activity of turmeric (Curcuma longa L.) extracts obtained using various techniques

Turmeric extracts were obtained from two lots of raw material (M and S) using various techniques: hydrodistillation, low pressure solvent extraction, Soxhlet, and supercritical extraction using carbon dioxide and cosolvents. The solvents and cosolvents tested were ethanol, isopropyl alcohol, and their mixture in equal proportions. The composition of the extracts was determined by gas chromatography-flame ionization detection (GC-FID) and UV. The largest yield (27%, weight) was obtained in the Soxhlet extraction (turmeric (S), ethanol = 1:100); the lowest yield was detected in the hydrodistillation process (2.1%). For the supercritical extraction, the best cosolvent was a mixture of ethanol and isopropyl alcohol. Sixty percent of the light fraction of the extracts consisted of ar-turmerone, (Z)-gamma-atlantone, and (E)-gamma-atlantone, except for the Soxhlet extracts (1:100, ethanol), for which only ar-turmeronol and (Z)-alpha-atlantone were detected. The maximum amount of curcuminoids (8.43%) was obtained using Soxhlet extraction (ethanol/isopropyl alcohol). The Soxhlet and low pressure extract exhibited the strongest antioxidant activities.     

Development and validation of an ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M1 in milk

A fast and ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M(1) in milk samples has been developed and validated. The method is an indirect competitive type format involving the immobilization of an aflatoxin M(1)-bovine serum albumin conjugate on 384 well black polystyrene microtiter plates and the use of a secondary antibody labeled with horseradish peroxidase detected with a luminol-based substrate. Aflatoxin M(1) standard solutions were prepared in milk-based buffer, and milk samples were analyzed without any cleanup procedure. The limit of quantification was 1 ppt, the coefficient of variation was below 9% for both intra- and interassay precision, and the recovery ranged from 96 to 122%. The method is specific, and other aflatoxins do not significantly cross-react with the antibody. Twenty-four milk samples were analyzed, and a good correlation was observed (y = 0.98x + 1.71, r(2) = 0.98, n = 24) when the data were compared with a reference high-performance liquid chromatography method with a fluorescent detector. The developed method is suitable for an accurate, sensitive, and high-throughput screening of aflatoxin M(1) in milk samples with a reduction of costs and increased detectability, as compared with previously developed immunoassays.     

The effects of grazing management in montado fragmentation and heterogeneity

The Portuguese silvo-pastoral system montado is broadly classified as a High Nature Value (HNV) system since it corresponds to farmland hosting high biodiversity levels, and such biodiversity depends on specific land use practices. However, in recent decades a decline both in the total montado area and in the tree cover density within the montado has been observed, driven mainly by management changes. This decline may result in biodiversity loss. Grazing is a central aspect determining the long-term sustainability of the montado system and it has implications also on the montado structural diversity, particularly on connectivity and heterogeneity, which is crucial for the maintenance of montado HNV. The aim of this paper is to demonstrate how variations in montado structural diversity are correlated with grazing management and its implications on the value of the system for conservation. The empirical data derives from a case study composed of 41 montado farms in two municipalities of the Alentejo region. Data on grazing management, biophysical and spatial factors were collected and several metrics were calculated to assess montado fragmentation and heterogeneity. A multivariate analysis was performed using generalized additive models. Results show that different grazing patterns, depending on stocking density and grazing animal type, are correlated with variations in montado fragmentation and heterogeneity. Particularly, cattle’s grazing is shown to have adverse effects on the montado fragmentation, while sheep grazing is shown to have stronger impacts on the heterogeneity within the montado patches.     

Seroepidemiological and clinical survey of feline immunodeficiency virus infection in northern Italy

Four hundred and thirty-nine feline serum samples from cats with different living conditions in the north of Italy were tested for antibodies to feline immunodeficiency virus (FIV) and for antigen of Feline Leukemia Virus by enzyme-linked immunosorbent assay. A Western blot technique was also used on the positive sera in order to confirm the presence of specific antibodies to FIV. The Western blot enabled the detection of a false positive serum. The prevalence of FIV infection in this population was 12.5% and among the seropositive cats a greater proportion was male (74.5%) than female (25.5%). A correlation between the clinical status and the evolution of the pathology is described together with a score based on the severity of the stomatitis in infected cats. The Western blot patterns of positive samples were then compared with the stage of the pathology. Statistical analysis on the distribution of FIV in stray cats, cats with garden and courtyard access and strictly house-confined cats showed a highly significant risk of the infection in the first group.     

Genetic complexity and multiple infections with more Parvovirus species in naturally infected cats

ABSTRACT: Parvoviruses of carnivores include three closely related autonomous parvoviruses: canine parvovirus (CPV), feline panleukopenia virus (FPV) and mink enteritis virus (MEV). These viruses cause a variety of serious diseases, especially in young patients, since they have a remarkable predilection for replication in rapidly dividing cells. FPV is not the only parvovirus species which infects cats; in addition to MEV, the new variants of canine parvovirus, CPV-2a, 2b and 2c have also penetrated the feline host-range, and they are able to infect and replicate in cats, causing diseases indistinguishable from feline panleukopenia. Furthermore, as cats are susceptible to both CPV-2 and FPV viruses, superinfection and co-infection with multiple parvovirus strains may occur, potentially facilitating recombination and high genetic heterogeneity. In the light of the importance of cats as a potential source of genetic diversity for parvoviruses and, since feline panleukopenia virus has re-emerged as a major cause of mortality in felines, the present study has explored the molecular characteristics of parvovirus strains circulating in cat populations. The most significant findings reported in this study were (a) the detection of mixed infection FPV/CPV with the presence of one parvovirus variant which is a true intermediate between FPV/CPV and (b) the quasispecies cloud size of one CPV sample variant 2c. In conclusion, this study provides new important results about the evolutionary dynamics of CPV infections in cats, showing that CPV has presumably started a new process of readaptation in feline hosts.     

Influence of deep frying on the unsaponifiable fraction of vegetable edible oils enriched with natural antioxidants

The influence of deep frying, mimicked by 20 heating cycles at 180 °C (each cycle from ambient temperature to 180 °C maintained for 5 min), on the unsaponifiable fraction of vegetable edible oils represented by three characteristic families of compounds (namely, phytosterols, aliphatic alcohols, and triterpenic compounds) has been studied. The target oils were extra virgin olive oil (with intrinsic content of phenolic antioxidants), refined sunflower oil enriched with antioxidant phenolic compounds isolated from olive pomace, refined sunflower oil enriched with an autoxidation inhibitor (dimethylpolysiloxane), and refined sunflower oil without enrichment. Monitoring of the target analytes as a function of both heating cycle and the presence of natural antioxidants was also evaluated by comparison of the profiles after each heating cycle. Identification and quantitation of the target compounds were performed by gas cromatography-mass spectrometry in single ion monitoring mode. Analysis of the heated oils revealed that the addition of natural antioxidants could be an excellent strategy to decrease degradation of lipidic components of the unsaponifiable fraction with the consequent improvement of stability.