Detection of Helicobacter spp. in gastric, fecal and saliva samples from swine affected by gastric ulceration

The aim of this study was to evaluate the presence of Helicobacter (H.) spp. in swine affected by gastric ulceration. Stomachs from 400 regularly slaughtered swine were subjected to gross pathological examination to evaluate the presence of gastric ulcers. Sixty-five samples collected from ulcerated pars esophagea and 15 samples from non-ulcerated pyloric portions were submitted to histopathological and molecular analyses, to detect Helicobacter spp., H. suis and H. pylori by PCR. Feces and saliva swabs were also collected from 25 animals in order to detect in vivo the presence of Helicobacter spp.. Gastric ulcers were detected in 373 cases (93%). The presence of ulcers in association with inflammatory processes was further confirmed by histological examination. Forty-nine percent (32/65) of the ulcerated esophageal portions as well as 53% (8/15) of the non-ulcerated pyloric portions were positive for Helicobacter spp. by PCR. The Helicobacter spp. positive samples were also positive for H. suis, while H. pylori was not detected. These results were confirmed by restriction enzyme analysis. With regard to feces and saliva samples, 15/25 (60%) and 16/25 (64%) were positive for Helicobacter spp. PCR, respectively but all were negative in H. suis and H. pylori specific PCR.     

The effect of climate on the presence of Culicoides imicola in Italy

A model was developed to classify the Italian territories in relation to their suitability to harbour populations of Culicoides imicola and, as a consequence, also able to sustain a bluetongue (BT) epidemic. Italy was subdivided into 3507 10 x 10 km cells. In 546 cells at least one collection was made. The cell was considered the unit for all subsequent analyses. Culicoides were collected using Onderstepoort-type blacklight traps. Some traps were operated weekly at chosen sites; the remainder were moved almost daily to new sites. Only the results obtained during the peak August-November period were used, to exclude bias caused by the seasonality of C. imicola. Climate data for the period 1999-2001 were obtained from 80 weather stations. Multiple logistic regression was performed using the presence or absence of C. imicola in a specific cell as the dependent variable. Annual means of daily values for minimum temperature and minimum relative humidity, and the mean altitude above sea level, were the independent variables. The probability of occurrence of C. imicola in each grid cell was used to create a prediction map for Italy. The model was able to correctly classify 77.5% of the 546 grid cells in which at least one collection had been made. Culicoides imicola was found frequently through much of Sardinia, in parts of southern Italy, and further north along the Tyrrhenian coast, but was absent from along most of the Adriatic coast, and the internal mainland, and from most of Sicily. Six detailed maps are provided. Also mapped are areas where the probability of the occurrence of C. imicola is lower than 5%. This identification of possible mountainous C. imicola-free areas in central Italy could facilitate safer animal trade and transhumance, even if BT infections in traded animals or moving stock, were to go undetected. Needless to say this depends upon no cool-adapted species of Culicoides being involved in the transmission of BT disease.     

Pollen morphology as fertility predictor in hybrid tea roses

Fertility of hybrid tea roses is often reduced due to their interspecific origin but also to intensive inbreeding. New genotypes used as pollen donors represent an economic risk for a breeding programme, as their influence on seed production is unknown. In this study 11 cut rose genotypes were selected from a company database as high fertile or low fertile male parents, according to the number of seeds per hybridisation. Pollen morphology and in vitro germination of the selected genotypes were characterised. Pollen was either small (mean diameter <30 μm), shrunken, and irregular (abnormal), or large (mean diameter >30 μm), elliptical and crossed by furrows (normal). High correlations were found between the number of seeds produced per hybridisation and the pollen diameter (r = 0.94) or the percentage of normal pollen (r = 0.96). In order to evaluate the predictive power of the models, we conducted regression analyses and performed a validation experiment on genotypes not present in the database and without background information on fertility. Pollen diameter and percentage of normal pollen were characterised and fitted in the regression models for seed set predictions. Validation with an independent dataset gave a good prediction for 83.3% of the data. This indicates that using either the mean pollen diameter or the percentage of normal pollen resulted in effective fertility prediction. This tool could enhance the genetic variability in crossings between hybrid tea roses, thus creating possibilities for less economically risky exploitation of new tetraploid genotypes as male parents.     

Use of innovative methods in the eradication of bovine tuberculosis

During final phases of eradication programmes, strains of Mycobacterium sp. not belonging to the tuberculosis complex increase their relative frequency and are responsible for positive skin test reactions. Moreover, the specificity of any indirect diagnostic test, such as the skin test, is never completely accurate, therefore even when tuberculosis infection is completely eradicated, a number of false positive reactions are to be expected. The aim of this paper is to evaluate the performances of traditional isolation/typing techniques, automatic isolation/typing techniques based on fluorimetric detection of bacterial growth (Bactec), skin tests and the -interferon test. Samples examined for the evaluation of test sensitivities originated from 154 infected animals belonging to 32 infected herds. Samples used as negative controls in the evaluation of test specificities originated from 86 animals of nine officially infection-free herds. The automatic isolation/typing technique based on fluorimetric detection of bacterial growth showed higher sensitivity than the traditional isolation typing technique. Moreover, it allowed a safer processing of bacterial cultures, decreasing the risk for laboratory workers. The observed performance of the gamma-interferon test was considered beneficial in that it increased the sensitivity of individual diagnosis within an infected herd, especially in 'problem herds', but its poor specificity did not improve detection of infected herds compared to the skin test.     

The contribution of stem cells to epidermal and hair follicle tumours in the dog

Background –  Although cutaneous stem cells have been implicated in skin tumourigenesis in humans, no studies have been conducted to elucidate the presence and the possible role of stem cells in hair follicle tumours in the dog. Hypothesis –  Stem cell markers are expressed in canine epidermal and follicular tumours and can be used to better understand the biology and origin of these tumours. Animals and Methods –  In the present study, normal skin sections and 44 follicular tumours were retrospectively investigated for the immunohistochemical expression of keratin 15 (K15) and nestin. In addition, 30 squamous cell carcinomas were evaluated for K15 expression. Results –  In normal skin, K15 and nestin were expressed in the outer root sheath cells of the isthmic portion of the hair follicle (bulge region), and K15 expression was also scattered in the basal cell layer of the epidermis. Infundibular keratinizing acanthomas, pilomatricomas and squamous cell carcinomas were mostly negative for K15, trichoblastomas were moderately to strongly positive, tricholemmomas were either negative or strongly positive, and trichoepitheliomas had heterogeneous staining. Nestin expression was generally faint in all follicular tumours. Conclusions and clinical importance –  Our results show that K15 can be a reliable marker for investigating the role of stem cells in hair follicle tumours of the dog, while nestin was judged to be a nonoptimal marker. Furthermore, our study suggests that hair follicle stem cells are present in the bulge region of hair follicles and could possibly play a role in tumourigenesis of canine tumours originating from this portion of the follicle, namely trichoblastomas, tricholemmomas and trichoepitheliomas. The loss of K15 expression in squamous cell carcinomas compared with normal skin suggests that this event could be important in the malignant transformation.     

Physiological host range of Trissolcus mitsukurii,a candidate biological control agent of Halyomorpha halys in Europe

Journal of Pest Science - The invasive stink bug Halyomorpha halys, native to East Asia, is a severe agricultural pest of worldwide importance, and chemical insecticides are largely sprayed for its...     

West Nile Virus Transmission in 2008 in North‐Eastern Italy

After 10 years, West Nile virus (WNV) re‐emerged in Italy in August 2008. As on 31 December 2008, the infection affected eight Provinces in three Regions (Emilia Romagna, Veneto, Lombardy), where a total of 794 cases of WNV infection in 251 equine stables were detected on the basis of the clinical signs and as a result of a serological screening in horses living in the area. Only 4.0% (32/794) of the serologically positive animals showed clinical signs, and the 32 clinical cases were reported in 18 different farms. The observed case‐fatality rate was 15.6% (5/32). The confirmed clinical cases were detected from end August to mid October. Significant levels of positivity by RT‐PCR were also observed in magpies (Pica pica) (9.1%, 95% confidence levels: 6.1–13.4%), carrion crows (Corvus corone) (7.4%, 95% confidence levels: 3.6–14.4%) and rock pigeons (Columba livia) (12.9%, 95% confidence levels: 7.6–21.2%).     

The use of homologous antigen in the serological diagnosis of brucellosis caused by Brucella melitensis

In the European Union the serological diagnosis of brucellosis caused by Brucella melitensis is performed using the heterologous antigen of B. abortus S99. The possible higher sensitivity or ability of an early detection of antibodies by a homologous antigen may prove very useful in the final phases of an eradication programme. Results obtained in sheep experimentally infected by B. melitensis biovar 3 were compared using B. abortus S99, B. melitensis M1, M2 and M3 antigens in the Rose Bengal plate test (RBPT), the complement fixation test (CFT) and an enzyme-linked immunosorbent assay (ELISA) test. Forty-six sheep from an officially brucellosis-free flock were experimentally infected intraconjunctivally with B. melitensis biovar 3. Prior to infection, all animals were tested first against Brucella antibodies, weekly for 2 months post-infection (PI) and then monthly for a further 7 months. All sera were tested against the antigens listed above using RBPT, CFT and ELISA. Using a Bayesian approach, test sensitivities were estimated and compared. Their ability for the early detection of antibodies was evaluated through a regression model based on a logit response model, using the number of days PI as the independent variable and the logit of the fraction of positive animals as the dependent variable. No significant differences were detected among the various antigens used, either in terms of sensitivity or in terms of antibody kinetics; however, the CFT was significantly less sensitive than the RBPT and ELISA and it also showed a lower rate of increase of percentage positive animals (beta-coefficient of regression analysis).     

Comparison of BSE prevalence estimates from EU countries for the period July to December 2001 to the OIE and EU GBR classifications

Consequent upon the bovine spongiform encephalopathy (BSE) crisis, the European Union (EU) Commission enacted various decisions, which demanded that all bovine animals over 30 months of age should be examined by one of the approved rapid tests when slaughtered for human consumption. All cattle over 24 months of age subject to 'special emergency slaughtering' or died on the farm or in transit or suspect of BSE infection should also be examined by one of the approved rapid tests. According to a specific commission decision, Sweden and Finland were to test only a sample of bovine animals over 30 months of age subject to normal slaughter. Testing commenced on 1 January 2001. The authors evaluate the results of more than 5 million tests performed in the second semester 2001 from across the EU. The prevalence of BSE in the risk categories considered (emergency slaughter, fallen stock and healthy slaughtered), and the probability distribution of true-positive, false-positive and false-negative results are estimated by second-order Bayesian analysis. The results of the validation of tests performed in the EU are also considered by estimation of the probability distribution of their sensitivity and specificity. The prevalence of infection estimated in the cattle population of each EU country is compared against the criteria given in the OIE Terrestrial Animal Health Code and is also used to evaluate the consistency of the results of EU Geographical BSE Risk with the actual infection levels in the countries. Finally, the capability of the two current approaches to BSE surveillance (i.e. the testing of all slaughtered and dead cattle as applied in the EU and a surveillance system targeted at animals in risk categories only) to detect the infection in a given population are discussed.