Application of fuzzy logic for the simulation of Plasmopara viticola using agrometeorological variables*

A mechanistic model called PLASMO was developed earlier to simulate grapevine downy mildew (Plasmopara viticola) and has been applied in several viticultural areas of Italy since 1988 by the collaboration of several research institutions of Firenze. In this study, a new simulation model based on fuzzy logic has been developed for the same structure (biological cycle of P. viticola). This approach allows classical quantitative information to be used together with qualitative information. Vague concepts can also be handled. Agrometeorological data is used, with an hourly time step, starting from budbreak to the end of the growing season. Air temperature, relative humidity, rainfall and leaf wetness are required. The simulated processes are the growth of grapevine leaf area and the main phases of the biological cycle of the pathogen: incubation, sporulation, germination, spore survival and inoculation. The main epidemiological outputs are timing of infection events and disease intensity. The performance of the model is evaluated and the mechanistic and fuzzy logic approaches are compared.     

B-mode ultrasound examination of canine mammary gland neoplastic lesions of small size (diameter < 2 cm)

Ultrasonography is a valuable tool for the evaluation of neoplastic lesions in the dog and there is a growing interest in the use of this technique for the stadiation of canine mammary tumours. An accurate assessment of small sized nodules facilitates the stadiation of the mammary lesions and helps the clinician in the choice of the most indicated surgical therapy. The aim of this study was to identify those ultrasound criteria that may be useful in discriminating between benign and malignant lesions of small size (diameter smaller than 2 cm). Sixty-two nodules, < 2 cm in larger diameter, belonging to thirty-five bitches presented between January 2012 and February 2014 were evaluated. Tumours were observed by conventional ultrasound and assessed for: shape (regular-irregular), limit (defined-ill-defined), margins (regular-irregular), echogenicity (hypoechoic-isoechoic-hyperecoic), echotexture (homogeneus-heterogeneus), presence of hyperecoic halo, distal acoustic enhancement or shadowing and surrounding tissue alterations. Among the alterations in surrounding tissues, the disruption of the glandular tissue and the increase in echogenicity of the peritumoral tissues were assessed. Thereafter, bitches were subjected to mastectomy and nodules were evaluated histologically. None of the ultasound criteria considered in the current study showed a statistically significant relation with malignancy, except for the presence of alterations in the tissue surrounding the nodules. According to our results, this characteristic may indicate malignancy, however its subjectivity may affect the applicability in clinical practice. In conclusions, conventional ultrasound in bitches had a limited ability in discriminating benign and malignant mammary gland neoplastic lesions of small size (diameter?<?2 cm).     

The influence of progesterone and prostaglandin F2α on decorin and the adhesion of caruncular epithelial cells of bovine placenta at early-mid pregnancy—Part II

One of the most important processes determining the proper course of gestation and its physiological termination in cows is the adhesion of epithelial cells allowing for direct contact of maternal and foetal parts of the placenta. Throughout pregnancy, placental cells are under strict hormonal control, which among others regulates the concentration and activity of specific proteins participating in the extracellular matrix remodelling of foetal membranes. The aim of the study was to evaluate the influence of progesterone and prostaglandin F_2α on the adhesion of epithelial cells at early-mid pregnancy in cows. Additionally, the impact of selected hormones on anti-adhesive properties of decorin was evaluated. Caruncular epithelial cells were isolated from healthy cows during pregnancy, immediately after slaughter. Primary cell cultures derived from the 2nd and 4th month of gestation were used in the experiments. The viability of cells was assessed by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. The activity of metalloproteinases was confirmed by the metalloproteinase assay. Progesterone (10^–5 and 10^–7 mol/L) and prostaglandin F_2α (10^–4_, 10^–5 and 10^–7 mol/L) increased the viability of bovine caruncular epithelial cells in the 2nd month of pregnancy. The treatment with prostaglandin F_2α significantly reduced the number of adherent cells from the 2nd month of gestation at the doses of 10^–4 and 10^–5 mol/L. Both progesterone and prostaglandin F_2α were shown to have an effect of decorin resulting in both a decrease in metalloproteinase activity and an increase in adhesion of cells to fibronectin.     

IMAGING DIAGNOSIS—URETHROVAGINAL FISTULA CAUSED BY A MIGRATING GRASS AWN IN THE VAGINA

A young intact female dog was presented with urinary incontinence. Abdominal ultrasound revealed the presence of hyperechoic linear structures within the cranial vagina suggestive of foreign material. A computed tomography (CT) retrograde vaginourethrogram demonstrated the presence of a fistulous tract between the urethra and vagina. A presumptive diagnosis of urethrovaginal fistula due to migration of foreign material was made. The grass awn was removed with vaginoscopic‐guided retrieval. Fourteen days later, surgical repair of the fistula and an ovariohysterectomy were done. This case report emphasizes the usefulness of CT for diagnosis and precise anatomical localization of genitourinary tract fistulas.     

Jaagsiekte sheep retrovirus found in milk macrophages but not in milk lymphocytes or mammary gland epithelia of naturally infected sheep

Jaagsiekte sheep retrovirus (JSRV) causes ovine pulmonary adenocarcinoma. JSRV can be transmitted via infected colostrum or milk, which contain somatic cells (SCs) harboring JSRV provirus. Nevertheless, the cell types involved in this form of transmission and the involvement of the mammary gland remain unknown. We separated adherent cells (macrophages and monocytes) by plastic adherence, and lymphocytes (CD4+ and CD8+ T cells, and B cells) by flow cytometry, from SCs in milk samples from 12 naturally infected, PCR blood test JSRV–positive, subclinical ewes. These cell populations were tested by PCR to detect JSRV provirus. The ewes were euthanized, and mammary gland samples were analyzed immunohistochemically to detect JSRV surface protein. We did not detect JSRV provirus in any milk lymphocyte population, but milk adherent cells were positive in 3 of 12 sheep, suggesting a potential major role of this population in the lactogenic transmission of JSRV. Immunohistochemistry did not reveal positive results in mammary epithelial cells, pointing to a lack of participation of the mammary gland in the biological cycle of JSRV and reducing the probability of excretion of free viral particles in colostrum or milk.     

Early flower initiation allows ample manipulation of flowering time in cherimoya (Annona cherimola Mill.)

Flower initiation date and readiness to flowering in buds of different age were studied in ‘Fino de Jete’ cherimoya (Annona cherimola) cultivar in order to establish the limits for the manipulation of its flowering date. Flower initiation was analyzed by light and scanning electron microscopy (SEM) collecting axillary buds from May to the following February, whereas the bud readiness to produce perfect flowers was determined by forcing buds of different age to sprout by means of leaf removal and tipping the new growth. SEM images confirm that cherimoya buds are differentiated into flowers almost a year before blooming. In this regard, axillary buds have already formed the sepals when the subtending leaf has just begun unfolding (week 0), while the petals are clearly visible in 1-week-old buds. Sectioning of paraffin-embedded buds illustrate that cherimoya buds are in fact a bud complex that 1 week after its inception comprises 4–5 buds of different size of which the two largest ones are reproductive, while the 2–3 smallest buds often remain undifferentiated at that time. The high capacity of flowering expressed by young buds that have been forced to grow proves that cherimoya meristems are early competent for flowering. No differences in fertility or in the time needed to reach anthesis after leaf removal were found among buds of different ages. Node position had no effect on bud break and flowering potential. The early flower initiation in cherimoya deduced from this work opens a wide temporal window for the experimental manipulation of flowering and harvest dates in this crop.     

Fatty acid profiles, tocopherol contents, and antioxidant activities of heartnut (Juglans ailanthifolia Var. cordiformis) and Persian walnut (Juglans regia L.)

The fatty acid and tocopherol compositions of three heartnut (Juglans ailanthifolia var. cordiformis) varieties (Imshu, Campbell CW1, and Campbell CW3) were examined and compared with those of two Persian walnut (Juglans regia L.) varieties (Combe and Lake). The major fatty acids found in heartnuts and walnuts were identified by gas chromatography as linoleic (18:2n-6), alpha-linolenic (18:3n-3), oleic (18:1n-9), palmitic (16:0), and stearic acid (18:0). Polyunsaturated fatty acids were the main group of fatty acids found in both heartnut and walnut, ranging from 73.07 to 80.98%, and were significantly higher in heartnut than in Persian walnuts (P < 0.001). In addition, heartnuts had significantly higher levels of 18:2n-6 and lower levels of 18:3n-3 compared to the Persian walnuts. gamma-Tocopherol was the main tocopherol homologue present in both types of nuts, followed by delta- and alpha-tocopherol. The highest concentration of gamma-tocopherol was found in Combe Persian walnut at 267.87 mug/g, followed by Lake Persian walnut and Imshu, Campbell CW1, and CW3 heartnut at 205.45, 187.33, 161.84, and 126.46 mug/g, respectively. Tocopherols, particularly the gamma-tocopherol, were found to contribute the most to the strong total antioxidant activities of both walnut and heartnut oils using either the free radical 2,2-diphenyl-1-picrylhydrazyl assay or the photochemiluminescence method.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.