CABI/EPPO distribution maps of plant pests and plant diseases and their important role in plant quarantine

Never before has the need for accurate distribution data for plant pests been so important. CAB International (CABI) and EPPO are international organizations with a long history and strong involvement in collating and disseminating information on the global distribution of plant pests. Distribution Maps of Plant Pests and Distribution Maps of Plant Diseases, first published in 1951 and 1942, respectively, are respected, referenced sources of such data, expertly compiled and validated and used by plant health organizations around the world. They have been joint CABI/EPPO publications since 1997, and provide an essential complement to expanding knowledge on plant pest distribution. The Distribution Maps continue to be the most authoritative sources of information on the presence and extent of specific plant pests, sourced from the 4.5 million records in CAB Abstracts as a basis and numerous other sources. They also feed directly into CABI's Crop Protection Compendium (CPC) and EPPO's Plant Quarantine Data Retrieval System (PQR) databases. Their history, compilation and value are discussed.     

Investigation of Senescence in Canine Fibroblasts

Introduction and Aims:  Primary and secondary cell cultures of canine dermal fibroblasts were used to test the suitability of several senescence-linked cellular markers to characterize the chronological age of dogs. In further studies these markers will be used to test the hypothesis that senescence-linked changes at the cellular level are representative for the observed inverse relationship of body mass and life expectancy in different dog breeds.
Methods:  In this study standardized skin samples of Beagles were used to analyse the cell morphology and several senescence-linked parameters as Cumulative Population Doubling Level, Highest Population Doubling Level, Culture Doubling Time and Life Span Completed to determine the replicative potential of fibroblasts. The samples were divided by the age of the donors into three groups ('young', 'middle' and 'old').
Results:  There are notable differences between the replicative potential of young and old Beagles at the cellular level. Preliminary results show that in fibroblast cultures obtained from the young group the highest population doubling level is higher and the culture doubling time is shorter then in cultures of the old group. Cultures from the young and middle-aged group cannot be distinguished by the measured parameters. For final results the complete statistical analysis of sampled data has to be carried out.
Conclusions:  Our results demonstrate a relationship between the chronological age of the skin donor and the replicative potential of the dermal fibroblasts. Therefore cellular senescence-linked markers could serve as suitable tools to compare the age of individual dogs of different breeds.     

Computed Tomography of the Lungs of the Dog by a Six-generation CT Scanner, Intravenous Contrast Medium and Different Windows

Computed tomography (CT) is a modern technique of image diagnosis particularly recommended in human medicine to evaluate the existence of pulmonary pathological changes such as neoplasms, metastasis, interstitial infiltrates, etc. In veterinary medicine, however, few anatomical and clinical CT studies in the dog have used apparatus of the latest generation, including injection of intravenous contrast and application of regional specific CT windows with different window width (WW) and window level (WL) to evaluate the lungs, the pulmonary vessels and the bronchial structures. This methodology allows the obtaining of clear CT images with high capacity of tissue discrimination and different shades of attenuation. In this work we have planned a tomographic study of the lungs of the dog by using a six-generation spiral CT scanner (Toshiba Ex Vision), belonging to the private Medical Institute of Radiology 'Irion' of Porto Alegre, Brazil. Four mixed-breed mature dogs (4–6 years, 15–20 kg) were used, two males and two females. The dogs were anaesthetized and kept in a maximum inspiration when obtaining the images. Dogs were placed in a stretcher in a ventral or sternal recumbency. Previously, the contrast urografin® was injected in the cephalic vein. Different CT windows were applied in order to increase the quality of the images: pulmonary window (WW 928; WL -680), high-resolution pulmonary window (WW 1085; WL -750), and soft tissue window (WW 652; WL -34). The use of intravenous contrast, different CT windows and a modern CT apparatus produced excellent images of the pulmonary parenchyma, the pleural cavity, the pulmonary veins, the lobular rami of the pulmonary artery and the lobular bronchi.     

An improved protocol for the production of AFLPTM markers in complex genomes by means of capillary electrophoresis

The amplified fragment‐length polymorphism (AFLP) technology is a recently introduced method to investigate genomes of different complexity, from microbial to higher organisms. It is applied to purposes as diverse as identification of species, strain and varieties, investigation of genetic diversity within and between populations, simple and complex trait mapping, and construction of linkage and physical maps. This technology has been designed on the use of primers labelled with radioactivity and on AFLP fragment separation on sequencing gel. We show that the original EcoRI/TaqI AFLP protocol does not perform appropriately when transferred to fluorescent labelling and capillary electrophoresis (CE), and propose an improved protocol for the production of high‐quality AFLP markers in fish, rodents and artiodactyles by means of the Beckman‐Coulter CEQ2000 automatic DNA sequencer. In addition, we describe the procedure routinely used in our laboratory to obtain binary matrices from AFLP profiles with the aid of Genographer free‐share software (vers. 1.6.0, J.J. Benham, Montana State University), able to elaborate original fragment data and convert them to standard graphical formats for phylogenetic analyses. Comparison with radioactive AFLPs in goats confirmed the reliability of the protocol developed for CE. In fact, 107 fragments generated by two primer combinations and identified by both techniques were attributed the same scoring. Compared with traditional methods, the use of capillary systems and automated analysis increases data throughput and scoring reliability, decreasing the overall experimental error.     

The coding sequence of the ASIP gene is identical in nine wild-type coloured cattle breeds

The aim of this study was to ascertain the role of the Agouti signaling peptide (ASIP) gene coding region in the Agouti locus variation within wild‐type coat colour in cattle. We determined the Extension genotype in 241 individuals from six Spanish and three French brown cattle breeds representative of wild‐type coat variation. Polymerase chain reaction–single‐strand conformation polymorphism (PCR‐SSCP) analysis was carried out in individuals of each Extension genotypes within the same breed in an attempt to identify variants in the three coding exons of the ASIP gene. No SSCP variants were found. Results were confirmed by sequencing the coding exons of the ASIP gene in 20 individuals. Our results suggest that the ASIP coding region does not play a central role in coat colour variation in cattle.     

Quantitative analysis of production traits in saltwater crocodiles (Crocodylus porosus): II. age at slaughter

Crocodile morphometric (head, snout‐vent and total length) measurements were recorded at three stages during the production chain: hatching, inventory [average age (±SE) is 265.1 ± 0.4 days] and slaughter (average age is 1037.8 ± 0.4 days). Crocodile skins are used for the manufacture of exclusive leather products, with the most common‐sized skin sold having 35–45 cm in belly width. One of the breeding objectives for inclusion into a multitrait genetic improvement programme for saltwater crocodiles is the time taken for a juvenile to reach this size or age at slaughter. A multivariate restricted maximum likelihood analysis provided (co)variance components for estimating the first published genetic parameter estimates for these traits. Heritability (±SE) estimates for the traits hatchling snout‐vent length, inventory head length and age at slaughter were 0.60 (0.15), 0.59 (0.12) and 0.40 (0.10) respectively. There were strong negative genetic (?0.81 ± 0.08) and phenotypic (?0.82 ± 0.02) correlations between age at slaughter and inventory head length.     

Commercial Seed Lots Exhibit Reduced Seed Dormancy in Comparison to Wild Seed Lots of Echinacea purpurea

Seed germination patterns were studied in E. purpurea (L.) Moench grouped by seed source, one group of seven lots from commercially cultivated populations and a second group of nine lots regenerated from ex situ conserved wild populations. Germination tests were conducted in a growth chamber in light (40 μmol·m(-2)·s(-1)) or darkness at 25 °C for 20 days after soaking the seeds in water for 10 minutes. Except for two seed lots from wild populations, better germination was observed for commercially cultivated populations in light (90% mean among seed lots, ranging from 82% to 95%) and in darkness (88% mean among seed lots, ranging from 82% to 97%) than for wild populations in light (56% mean among seed lots, ranging from 9% to 92%) or in darkness (37% mean among seed lots, ranging from 4% to 78%). No germination difference was measured between treatments in light and darkness in the commercially cultivated populations, but significant differences were noted for treatments among wild populations. These results suggest that repeated cycles of sowing seeds during cultivation without treatments for dormancy release resulted in reduced seed dormancy in E. purpurea.     

Effects of Clinical Mastitis on Ovarian Function in Post-partum Dairy Cows

Mastitis-induced ovarian abnormalities were studied in a field trial. At 1-3 day after calving, > or = 2 parity cows not affected with chronic recurrent mastitis and yielding < 400,000/ml somatic cell count (SCC) individual milk in the previous lactation, were enrolled in the study. Thereafter milk samples were collected three times weekly for 95-100 day for progesterone (P4) assay. Individual P4 profiles were used to monitor ovarian cyclicity. When mastitis was diagnosed in the first 80 day post-partum (pp), clinical signs were recorded and scored, and aseptic milk samples were taken to identify the mastitis pathogens. Depending on the isolated pathogens the cows were blocked into one of the three sub-groups affected by either Gram-positive (GP), or Gram-negative (GN) bacteria, or of those with no detected pathogens (NDP). Cows suffering from any type of mastitis between days 15 and 28 (n = 27) showed a delay in the onset of ovarian cyclicity, and estrus was postponed compared to cows affected during the first 14 day pp (n = 59) and controls (n = 175) (38.6 +/- 2.3 vs 33.4 +/- 2.1 and 32.0 +/- 1.0 day, respectively, for onset of ovarian cyclicity and 90.7 +/- 2.5 vs 80.2 +/- 2.8 and 83.9 +/- 2.1 day, respectively, for estrus; both p < 0.05). The percentage of cows ovulating by day 28 was lower in those affected by mastitis between days 14 and 28 compared to cows between days 1 and 14 and controls (22.2% vs 47.5 and 50.3%, respectively; p < 0.05). A significantly higher rate of premature luteolysis was observed in GN + NDP compared to GP mastitis and healthy cows (46.7% vs 8.3 and 2.0%, respectively; p < 0.001). If the mastitis outbreak occurred during the follicular phase, the duration of this cycle segment was lengthened in GN + NDP mastitis compared to GP mastitis and healthy cows (10.8 +/- 0.9 vs 7.9 +/- 0.1 and 7.2 +/- 0.1, respectively; p < 0.001). The results indicate that mastitis can affect the resumption of ovarian activity in pp dairy cows. Mastitis may also impair reproduction also in cyclic cows: this effect can be the consequence of premature luteolysis or a prolonged follicular phase.     

Relation between Physical Properties of the Zona Pellucida and Viability of Bovine Embryos after Slow-freezing and Vitrification

In vitro-produced bovine morulae/blastocyst embryos (n = 119) were slow-frozen and vitrified and the physical alterations of the zona pellucida (ZP) was observed by scanning electron microscopy (SEM) to find an explanation for the loss of developmental capacity of the embryos after freezing/thawing. A control group was provided, in which embryos (n = 38) were neither frozen nor vitrified. Embryos were in vitro-cultured in a standard CO2 Heraeus incubator and their viability was assessed 24 and 48 h after the start of culture, evaluating their morphological aspect. After 24 h of culture, embryo survival rate for slow-freezing/thawed (n = 23), vitrified/thawed (n = 20) and control embryos (n = 20) was 39, 27 and 90%, and 35, 14 and 65% after 48 h of culture, respectively. For evaluation of physical changes occurring in ZP, 20 embryos were slow-frozen, 18 were vitrified and 18 were used as control. All embryos were fixed, dried and examined under an SEM. Embryo's diameter, as well as the number of pores and their diameter was measured in squares of 6.4 microm width. We observed that, on average, the diameter of the embryos (92.26 +/- 10.15 microm) did not differ significantly among all embryos. As far as the diameter of the pores in the outer surface of the ZP is concerned, the results revealed a significant difference (P < 0.05) between control (0.48 +/- 0.0025 microm), slow-frozen (0.34 +/- 0.0007 microm) and vitrified (0.27 +/- 0.0006 microm) embryos. For the number of pores, statistical differences (p < 0.05) were observed between control and vitrified embryos (45.4 +/- 7.3 vs 38.2 +/- 8.2). It is possible that ZP functions as a barrier which is positive when dealing with pathogens, but is harmful when nutrients were supplied from the outside, especially at 48 h of culture. Results indicate that the steps of cryopreservation cause alterations in ZP, with irreversible damage on the further developmental competence of bovine embryos.