Oxidative stress in the pathogenesis of canine zinc‐responsive dermatosis

Zinc deficiency causes skin diseases both in humans and in animals. The underlying pathogenic mechanisms remain unclear, but a growing body of evidence indicates a role for zinc in skin protection against free radical‐induced oxidative damage. The immunohistochemical expression of heat shock proteins (HSPs; Hsp27, Hsp72, Hsp73 and Hsp90), Cu/Zn superoxide dismutase (SOD), metallothionein (MT), Ki‐67 antigen and active caspase‐3 were evaluated in normal canine skin and in samples from eight dogs with zinc‐responsive dermatosis. All investigated HSPs showed intense cytoplasmic immunostaining in the affected epidermis. Focal nuclear positivity of Hsp72 was also detected in keratinocytes. Although Cu/Zn SOD expression was similar to that observed in normal skin, MT immunoreactivity occurred in both the cytoplasm and the nucleus of basal cells in normal skin but was absent from the affected epidermis. Caspase‐3 activation was also absent in the involved epidermis, which revealed a high Ki‐67 index (a 3.5‐ to 9‐fold increase compared with normal skin). These results support the hypothesis that cellular response to stress, particularly oxidative stress, is involved in the pathogenesis of skin lesions in canine zinc‐responsive dermatosis. The lack of MT immunoreactivity in the affected epidermis may be indicative of low zinc levels, thus resulting in vulnerability to oxidative damage. In contrast, high expression levels of HSPs in skin during zinc deficiency may confer protection against a variety of dangerous stimuli, contributing to inhibition of apoptosis and to cell cycle regulation of proliferating keratinocytes.     

Paraneoplastic leukocytosis in a dog with a renal carcinoma

A 7-year-old male German Shepherd dog in poor body condition had a 3-month history of intermittent hematuria. Nonregenerative anemia, mild leukocytosis, marked hypoalbuminemia, and hematuria were observed. Subsequently, marked neutrophilia and moderate monocytosis were noted; anemia, hypoalbuminemia, and hematuria persisted; and the dog developed disseminated intravascular coagulation. Ultrasonographic examination of the abdomen revealed the presence of an enlarged and irregularly shaped right kidney with a large area of cavitation, and a nephrectomy was performed 30 days after initial examination. Cytologic examination of fine-needle aspirates and imprints of the right kidney revealed a neoplastic cell population suggestive of renal carcinoma. The histopathologic diagnosis was chromophobic cystic-papillary renal carcinoma. The tumor cells expressed granulocytic/macrophage-colony-stimulating factor (GM-CSF), detected by immunohistochemical staining, and elaboration of GM-CSF by the tumor may have mediated the leukocytosis in this dog. Following excision of the tumor, neutrophil and monocyte counts were only mildly increased. The dog died 135 days after initial presentation, and a necropsy was not permitted. Paraneoplastic neutrophilic leukocytosis is an uncommon finding and may be caused by elaboration of CSF by neoplastic cells.     

Effects of tetracycline and zinc on selection of methicillin-resistant Staphylococcus aureus (MRSA) sequence type 398 in pigs

An in vivo experiment was conducted to evaluate the effects of tetracycline and zinc on pig colonization and transmission of methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST) 398. Eight piglets naturally colonized with MRSA ST398 and 8 MRSA-negative piglets of the same age and breed were assigned to three groups treated with tetracycline and zinc (Group 1), zinc (Group 2) or tetracycline alone (Group 3) and one non-treated group (Group 4), each containing two MRSA-positive and two MRSA-negative animals. Two additional non-treated control groups composed of only MRSA-positive (Group 5) and MRSA-negative (Group 6) animals were used to check for stability of MRSA carriage status. Nasal swabs and environmental wipes were collected on Days 0, 7, 14, and 21, and the occurrence of MRSA in each sample was quantified by bacteriological counts on Brilliance? MRSA agar. Significantly higher nasal MRSA counts were observed in the zinc-treated (p=0.015) and tetracycline-treated (p=0.008) animals compared to the non-treated animals. Environmental MRSA counts appeared to increase over time in Groups 1 and 2 but such an increase was not statistically significant. MRSA-negative animals housed with MRSA-positive animals became positive in all groups, whereas the carriage status of the animals in Groups 5 and 6 did not change. This study demonstrates that feed supplemented with tetracycline or zinc increases the numbers of MRSA ST398 in the nasal cavity of pigs. Transmission of MRSA from positive to negative animals housed within the same pen was not influenced by exposure to these agents.     

One dose of a porcine circovirus 2 subunit vaccine induces humoral and cell-mediated immunity and protects against porcine circovirus-associated disease under field conditions

This study investigated the efficacy of a one-dose porcine circovirus 2 (PCV2) subunit vaccine based on the PCV2 Cap protein expressed in a baculovirus system on two different farms at which a history of porcine circovirus-associated disease (PCVD) was present. Morbidity, mortality, average daily weight gain, carcass weight, PCV2 load in serum and vaccine immunogenicity were assessed. Serology to porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae was performed. A double-blind, randomised, and controlled field trial was performed distributing 818 piglets between two treatment groups. At inclusion (weaning at 21 ± 3 days of age), 408 animals (group B) received a 2-mL intramuscular dose of Porcilis PCV(?) (vaccinated group). Controls (group A, 410 pigs) received 2 mL of the adjuvant Diluvac Forte(?) intramuscularly. Weights were recorded at inclusion and at 12 and 26 weeks of age, and the average daily weight gain (ADWG) was calculated. The carcass weights of the pigs from farm 2 were recorded at slaughter (274 days old). All dead animals (died or culled) underwent autopsy to classify them as PMWS-affected or not. At each farm, blood samples were taken from 22 pigs/group for serologic studies. A beneficial effect was found after vaccination with a single dose of a PCV2 Cap vaccine against PCVD. The vaccination reduced the mortality rate and morbidity, reduced PCV2 viremia and viral load, improved productive performances (e.g. ADWG: +70 g/day between 12 and 26 weeks of age when viremia and the specific disease occurred) as well as carcass weight at slaughter age (+4.5 kg). These effects were associated with virologic and clinical protection from the immunogenicity of the vaccine measured as activation of both a humoral and a cellular immune response.     

Molecular weight distributions and linkages in lignocellulosic materials derivatized from ionic liquid media

A novel and reproducible method is described for accurately determining the molecular weight distribution by size exclusion chromatography (SEC) of whole lignocellulosic materials. This approach offers the opportunity to compare the molecular weight distributions of intact milled woods and its component fractions, lignins and holocelluloses, all from the same source, thus highlighting the potential of the technique and the contributions of the individual components to the chromatogram. The method is based on the dissolution of the ball-milled samples in the ionic liquid 1-allyl-3-methylimidazolium chloride ([amim]Cl). Under these homogeneous ionic liquid media, a derivatization reaction was performed with benzoyl chloride in the presence of pyridine. The thoroughly benzoylated wood with its associated carbohydrate and lignin components was found to be completely soluble in the THF SEC eluent with marked UV detector sensitivity. This methodology, when applied to the individually isolated holocellulose and lignin (enzymatic mild acidolysis lignin; EMAL) materials from Norway spruce ( Eucalyptus grandis ) wood and corn stover, offered a better understanding as to the possible ways the lignin and the carbohydrates may interact within these three different species. Finally, the applicability of the methodology is shown for a series of pure cellulosic samples under intense mechanical defibration conditions, offering a visualization of the molecular weight distribution changes induced during the production of nanofibrillated cellulose.     

Evaluation of a commercial intravaginal thermometer to predict calving in a Hungarian Holstein-Friesian dairy farm

In this study, the utility of a commercial intravaginal thermometer was evaluated as an automated method for the prediction of calving in a total of 257 healthy pregnant Holstein–Friesian female cattle. The accuracy and the sensitivity of predicting calving within 48 hr before calving were also evaluated. The intravaginal temperature changes from 72 hr before and up to calving were significantly (p ≤ .001) affected by parity, season (summer vs. autumn), the time of day (8 a.m. or 8 p.m.) and the 6-hr time intervals (38.19°C: first interval 0 to 6 hr before calving vs. 38.78°C: twelfth interval 66 to 72 hr before calving), while the gender (p = .943), and the weight of the calf (p = .610), twinning (p = .300), gestation length (p = .186), foetal presentation (p = .123), dystocia (p = .197) and retention of foetal membranes (p = .253) did not affect it significantly. The sensitivity of the SMS of expecting calving within 48 hr and the positive predictive value were 62.4% and 75%, respectively, while the sensitivity and the positive predictive value for the SMS of expulsion reached 100%. It can be concluded that the investigated thermometer is not able to predict calving within 48 hr accurately; however, imminent calving can be accurately alerted.     

A natural fungicide for the control of <Emphasis Type="Italic">Erysiphe betae</Emphasis> and <Emphasis Type="Italic">Erysiphe cichoracearum</Emphasis>

This study examines the effects of a vegetable fungicide on sugar beet powdery mildew (Erysiphe betae) and cucumber powdery mildew (Erysiphe cichoracearum). The formulations consisting of a dispersion of Brassicaceae meal in vegetable or mineral oils on infected leaves of sugar beet, reared in the greenhouse, and of musk melons cultivated under plastic tunnels, were tested in comparison to each oil taken separately. Both formulations containing Brassicaceae meals, caused 94% of conidia to be distorted while for the untreated group only 2% were distorted. Furthermore, the leaf area infected by E. betae was 56% for untreated plants and 2.7 and 9.9% respectively, for plants treated with meal containing mineral and vegetable oil. Vegetable oil considered separately or with Brassicaceae meals showed no phytotoxicity, while the formulations based on mineral oil showed a significantly lower fresh and dry weight on tomato plants. The low level or absence of phytotoxicity of plants treated with vegetable oil formulations suggests that to improve the efficacy of powdery mildew control, they could be used mixed with sulphur. The efficiency of the vegetable formulations in the powdery mildew control observed during these trials encourages further investigation on other parasitic fungi and foliar pathogens.     

Volatile organic compounds (VOC) as biomarkers for detection of Ceratocystis platani

Ceratocystis platani causes canker stain of plane trees, and it represents a serious disease of Platanus spp. both in the United States and Europe. Current chemical or biological controls do not effectively manage C. platani, so new preventive methods need to be developed in order to limit this pathogen spreading. In this work, we have characterized the main volatile organic compounds (VOC) emitted in vitro from pure cultures of C. platani and other common pathogenic fungal species of hosts plants growing in the same ecosystems as plane trees. We found that C. platani emitted a similar blend of VOC compared with phylogenetically similar species C. populicola. In particular, C. platani was characterized by emission of isoamyl acetate and isobutyl acetate while C. populicola by ethyl acetate and isobutyl acetate, which were not released by any of the other out‐group fungal species grown on the same medium. Moreover, following a targeted approach based on the main VOC found in vitro, we have successfully validated in vivo that VOC uniquely emitted by C. platani (i.e. isobutyl acetate along with isoamyl alcohol) were released from the bark of plane trees following C. platani inoculation. Our results highlight the possibility to exploit VOC emitted specifically by C. platani as biomarkers to recognize Platanus x acerifolia plants infected by this pathogen.     

Adenocarcinoma of the disseminated prostate in a cat

An adenocarcinoma of the disseminated prostate gland with pulmonary, myocardial and renal metastases is described in a 12-year-old, neutered male European cat. Histologically, the tumour was localised in the spongy layer of the prostatic urethra and showed an epithelial alveolar pattern. Considering the anatomic, microscopic and immunohistochemical findings, the tumour was diagnosed as an adenocarcinoma of the disseminated prostate gland. To our knowledge this is the first report of adenocarcinoma of the disseminated prostate gland in a cat.