Conventional and Doppler ultrasound for the differentiation of benign and malignant canine mammary tumours

Objectives : The aim of this study was to evaluate the sensitivity of conventional and Doppler ultrasound for differentiation of benign and malignant mammary tumours in female dogs. Methods : Mammary tumours were evaluated from 60 animals and divided into two distinct groups, group 1 (benign tumours) and group 2 (malignant tumours). The tumours were assessed by conventional ultrasound, Doppler ultrasound mode, histopathology and immunohistochemical detection of vascular endothelial growth factor. Results : Conventional ultrasound examination was found to be ineffective in separating tumours into the two experimental groups. Similarly, using colour‐flow Doppler ultrasound, no correlation was found between the presence of vascularisation and its characteristics between the two groups. Triplex Doppler ultrasound yielded average maximum velocities of 28·71 cm/s for malignant and 19·91 cm/s for benign tumours, which were significantly different (P=0·01). For vascular endothelial growth factor, an average score of 2·22 was found for group 2 and 1·66 for group 1 (P=0·03). Positive correlations were found between vascular endothelial growth factor and presence of vascularisation (P=0·04 and r=0·3658) and between vascular endothelial growth factor and maximum velocity (P=0·03 and r=0·3913). Clinical Significance : Doppler evaluation may be used to predict malignancy of mammary tumours in bitches.     

Serosurvey of Aujeszky's disease virus infection in European wild boar in Spain

Serum samples from 693 hunted wild boar (Sus scrofa) were analysed by means of a blocking ELISA technique, and the mean (se) prevalence of antibodies to Aujeszky's disease virus was 44 (4) per cent. All the seropositive wild boar were from south central Spain, except for one from central Spain, close to the main positive area. In this area, where large game species are increasingly managed for hunting, the seroprevalence was affected by the type of management. More intensively managed populations had a higher prevalence than wild boar living in natural situations, and the seroprevalence increased with the age of the animals; the seroprevalence was higher in females in all age groups. The seroprevalence in males more than one year old peaked after the breeding season, whereas females of the same age had a higher and constant seroprevalence throughout the year.     

Molecular characterization of Mycobacterium tuberculosis complex isolates from wild ungulates in south-central Spain

The role of European wild ungulates in the epidemiology of tuberculosis (TB) is still under discussion. This study describes the geographical distribution and molecular typing of 77 Mycobacterium tuberculosis complex isolates belonging either to M. bovis or to M. caprae, cultivated from hunter harvested red deer (Cervus elaphus) and European wild boar (Sus scrofa) in 24 Spanish localities, and compares them with spoligotypes detected previously in humans, livestock or wild animals, as described in the literature. The distribution of the molecular type patterns suggests that the population of M. tuberculosis complex strains isolated from Spanish wild ungulates is spatially structured despite the lack of important geographical barriers and despite the increasingly frequent wildlife translocations. Red deer and the European wild boar can share the same molecular types in localities in which the M. tuberculosis complex was isolated from both species. Strains of bovine and caprine origin do circulate in the same local wildlife populations. Six out of 11 spoligotypes were similar to types described in human cases. The isolation of TB strains in fenced estates from wild animals that have not had contact with domestic livestock for at least the past two decades, strongly suggests that the M. tuberculosis complex is able to survive in these populations. Therefore, wildlife including cervids and the wild boar need to be considered in the epidemiology and control of tuberculosis.     

Wild boar helminths: risks in animal translocations

The helminth populations found in a group of wild boars collected in central Spain were compared to those in a group of animals imported from a French game farm that produces boars for restocking. Eleven helminth species, including ten nematodes and one acanthocephalan, were found. Gongylonema pulchrum and Macracanthorhynchus hirundinaceus were only detected in autochthonous wild boars, while Oesophagostomum dentatum, Ascaris suum, and Trichuris suis were detected in imported animals only. Autochthonous wild boars were more frequently and more intensely parasitised by Ascarops strongylina than the imported ones. No differences in prevalence nor intensity were found for the species Capillaria garfiai, Globocephalus urosubulatus, Metastrongylus sp., Physocephalus sexalatus and Simondsia paradoxa. To our knowledge, G. urosubulatus, G. pulchrum and S. paradoxa have not previously been described in wild boars in Spain. Our results highlight the risks of translocating wild animals, with regard to their helminth parasites. Until improved control measures are established, it would be wise to avoid long-distance translocations in order to prevent the potential introduction of foreign parasites.     

Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain

Cucurbit powdery mildew caused by Podosphaera fusca limits crop production in Spain. Since its management is strongly dependent on chemicals, the rational design of control programmes requires a good understanding of the fungicide resistance phenomenon in field populations. Fifty single-spore isolates of P. fusca were tested for sensitivity to three quinone-outside inhibiting (QoI) fungicides: azoxystrobin, kresoxim-methyl and trifloxystrobin. Minimum inhibitory concentration (MIC) values for QoI-sensitive isolates were found to range from 0.25 to 10 μg ml⁻¹ for azoxystrobin to 5–25 μg ml⁻¹ for kresoxim-methyl, using a leaf disc-based bioassay. High levels of cross-resistance to QoI fungicides were found. Eleven isolates showed resistance to the three QoI fungicides tested with MIC and EC₅₀ values >500 μg ml⁻¹ resulting in RF values as high as >715 and >1000 for trifloxystrobin and azoxystrobin, respectively. A survey of P. fusca QoI resistance was carried out in different provinces located in the south central area of Spain during the cucurbit growing seasons in 2002, 2003 and 2004. Examination of a collection of 250 isolates for QoI resistance revealed that 32% were resistant to the three fungicides tested; the provinces of Ciudad Real, Córdoba and Murcia being the locations with the highest frequencies of resistance (44–74%). By contrast, no resistance was found in Badajoz, and relatively low frequencies were observed in Almería and Valencia (10–13%). Nearly 50% of resistant isolates were collected from melon plants. Based on these data, recommendations about the use of QoI fungicides for cucurbit powdery mildew management in the sampled areas are made.     

Oocyte morphometry in female dogs (Canis familiaris, Linnaeus, 1758)

In the present study, the ovaries of 15 healthy bitches divided into three groups according to age were analysed by histology and morphometry: group 1 (1-3 years), group 2 (>3-5 years) and group 3 (>5-7 years). After ovariosalpingohysterectomy, the ovaries were fixed, routinely processed for embedding in paraffin and stained with haematoxylin-eosin. The following morphometric parameters were analysed: maximum and minimum diameter (microm), perimeter (microm), area (microm(2)) and roundness of the cytoplasm and nucleus of oocytes from different types of follicles. Significant differences in the cytoplasmic and nuclear parameters of follicular oocytes between the experimental groups were determined by anova and the Tukey test (5%). For the biotechnology of reproduction, the present results showed that in vitro maturation yielded the best performance for oocytes from primordial follicles in group 3 females and for oocytes embedded in secondary and tertiary follicles in group 2 females. In addition, the present findings will help in the diagnosis and prognosis of diseases affecting bitches, and will especially contribute to a better understanding of these cells by researchers in the field of histology and canine reproduction.     

RFLP variability in apricot (Prunus armeniaca L.)

The level of polymorphism of the restriction fragment length polymorphisms (RFLPs) detected by 33 almond genomic and cDNA probes was studied in a set of 52 European and North-American apricot cultivars. Eighteen of these probes were polymorphic and yielded a total of 48 scorable bands, allowing the identification of 45 different phenotypes. Most cultivars (43) had an individually distinguishable RFLP phenotype, and three of the five clusters with the same phenotype contained cultivars that were likely to be synonymous. The group of Spanish cultivars (25) had a lower level of polymorphism than the others, suggesting that bottlenecks may have occurred in the recent history of the apricot that have eroded its genetic variability.     

Free and conjugated polyamines and phenols in raw and alkaline-clarified sugarcane juices.

Sugarcane juice contains a lot of sucrose associated with several monosaccharides, defined as low molecular mass carbohydrates (LMMC), as well as some polysaccharides and glycoproteins, which are defined as mid and high molecular mass carbohydrates (MMMC and HMMC, respectively). These three categories of carbohydrates can be separated by size-exclusion chromatography through Sephadex G-10 and Sephadex G-50 columns, but elution profiles change drastically after juice clarification performed by adjusting the pH value of the juice to 8.0. In addition, polyamines and some phenolics are currently associated with carbohydrate preparations, and the distribution pattern of these conjugates also changes after clarification. Polyamine levels generally decrease after juice clarification. Cadaverine is completely removed from the different carbohydrate preparations, whereas spermidine is the main polyamine occurring in association with sugarcane carbohydrates, as free or acid-soluble form in LMMC preparation or as acid-soluble and -insoluble forms in both MMMC and HMMC preparations. Polyamines, presumably spermidine, conjugate to p-hydroxybenzoic acid in LMMC, mostly to caffeic acid in MMMC, and to syringic acid in HMMC preparations. HMMC-associated polyamines appear in both acid-soluble and -insoluble fractions. Syringic acid also occurs in the LMMC preparation, but juice clarification changes it from acid-soluble to free form, and it coelutes with sucrose.     

Morphological development and growth patterns of the leopard grouper Mycteroperca rosacea during larval development

At 25 °C, metamorphosis in leopard grouper Mycteroperca rosacea larvae took 60 days after hatching. The total length at day 1 was 1.95±0.22 mm and juveniles reached a length of 30.64±0.23 mm at day 60; the increase was approximately linear. We describe eight stages of development during this period. Larvae with the yolk sac attached occur from days 1 to 4 (Stages 1 and 2). The preflexion occurs on days 5–20 (Stages 3 and 4). Bending notochord occurred at day 25 (Stage 5). The other morphological changes that precede the juvenile phase occurred progressively until day 60 (Stages 6–8). Allometric growth in the height and length of the head, trunk length, height and length of the tail and the diameter of the eye compared with the total length showed two distinct stages of growth. Inflexion point, where growth is positive, occurred when larvae reached between 18.75 and 21.59 mm, which corresponds to larvae at days 35–40.     

Effects of dietary live yeast Debaryomyces hansenii on the immune and antioxidant system in juvenile leopard grouper Mycteroperca rosacea exposed to stress

This study investigated the effects of the dietary intake of live yeast Debaryomyces hansenii (CBS 8339) on the immune response of juvenile leopard groupers Mycteroperca rosacea and on their resistance to infection by dinoflagellates Amyloodinium ocellatum.During 4 weeks, juvenile groupers were fed either a diet supplemented with D. hansenii at 10⁶ cfu g^− 1 or a control diet. Both groups were then transferred to a system infected with A. ocellatum and maintained in this stressful environment, for one week. Rearing conditions were favorable to the proliferation of the dinoflagellate and, at 7 days post-infection, the cumulative mortality rate of the control group was 89.5%, suggesting that the yeast-enriched diet conferred protection against the parasite. Only fish with yeast-supplemented diet recovered from the A. ocellatum challenge.The immunological response of both groups of fish was evaluated. Samples were collected at weeks 4 (stressed fish) and 7 (recovering fish) to determine the levels of hemoglobin, plasmatic protein, and immunoglobulin M (IgM) as well as the activity of antioxidant enzymes (SOD and CAT). IgM increased significantly (P ≤ 0.05) in the recovering fish, which had all been fed the yeast diet. A significantly increase was also observed in superoxide dismutase activity in juvenile fish fed the yeast diet; however catalase activity was not affected by the dietary yeast supplement. Overall, fish that received the yeast supplement in their diet have an enhanced immune response as compared with their control counterparts. The present results show that a diet supplemented with D. hansenii stimulates the immune system of juvenile M. rosacea thus enhancing their resistance against A. ocellatum.