Evaluation of the cationic trypsinogen gene for potential mutations in miniature schnauzers with pancreatitis

The purpose of this study was to evaluate the cationic trypsinogen gene in miniature schnauzers for possible mutations. Genetic mutations have been linked with hereditary pancreatitis in humans. Four miniature schnauzers were selected on the basis of a clinical history of pancreatitis. One healthy miniature schnauzer and 1 healthy mixed breed canine were enrolled as controls. DNA was extracted from these canines using a commercial kit. Primers were designed to amplify the entire canine cationic trypsinogen cDNA sequence. A polymerase chain reaction (PCR) was performed and products were purified and sequenced. All sequences were then compared. The healthy control canine, a healthy miniature schnauzer, and the 4 miniature schnauzers with pancreatitis showed identical sequences of the cationic trypsinogen gene to the published sequence. We conclude that, in contrast to humans with hereditary pancreatitis, mutations of the cationic trypsinogen gene do not play a major role in the genesis of pancreatitis in the miniature schnauzer.     

Laboratory diagnosis and disease occurrence in the current African swine fever eradication program in Spain, 1989–1991

The purpose of this study was to determine the rate of decline and geographical distribution by municipality of clinical and subclinical African swine fever (ASF) in the affected areas of Spain. A second aim was to evaluate the performance of diagnostic tests in the Spanish ASF eradication program. Clinical outbreaks were confirmed using both the direct and indirect immunofluorescence test (and if both were negative, by the hemabsorption test). The serological status of swine was determined by an enzyme-linked immunosorbent assay (ELISA) and suspect serum samples were confirmed by the immunoblot assay.

The number of clinical outbreaks (herds) of ASF for 1989, 1990 and 1991 was 170, 347 and 207, respectively. The numbers of municipalities within each affected province experiencing acute outbreaks for the same time periods were 49, 69 and 48, respectively. Serologically diagnosed animals positive for ASF were 1.1% of animals tested in 1989, 0.5% in 1990 and 0.8% in 1991. The corresponding positive predictive values of the standard ELISA test used were 99.0, 97.9 and 98.8, respectively. Similarly, the number of municipalities within each affected province experiencing serologically positive subclinically infected animals was 269, 178 and 147 for each of the years 1989, 1990 and 1991, respectively.     

Review of methodologies and a protocol for the <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of wheat

Since the first report of wheat transformation by Agrobacterium tumefaciens in 1997, various factors that influence T-DNA delivery and regeneration in tissue culture have been further investigated and modified. This paper reviews the current methodology literature describing Agrobacterium transformation of wheat and provides a complete protocol that we have developed and used to produce over one hundred transgenic lines in both spring and winter wheat varieties.     

Effect of bleaching on lipid content and composition of Okinawan corals

Lipid contents and compositions of the bleached Okinawan corals were analyzed for the first time. Bleached corals collected at Sesoko Okinawa, Japan (26°38′N, 127°52′E), after the 1998 bleaching event showed decreased lipid content with a concomitant decline of wax in the lipid composition. The reductionin the lipid content of corals showed diversity depending on their colony morphology. On the whole, corals of massive morphology were relatively rich in lipid even after the bleaching event, and hence lesser reduction in the lipid content compared with the cases for those of branching morphology. Furthermore, there was a positive correlation between lipid content and zooxanthellae density in the bleached corals, suggesting again the importance of symbiont for the supply of lipid to the host cells, and hence for the survival of the bleaching event.     

Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus

In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease—Neutrase^®—was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection.     

Influence of pond fertilization and feeding rate on growth performance, economic returns and water quality in a small-scale cage-cum-pond integrated system for production of Nile tilapia (Oreochromis niloticus L.)

The effects of pond fertilization and feeding rate on growth, economic returns and water quality were investigated to develop a low‐cost cage‐cum‐pond integrated system for production of Oreochromis niloticus (L.). Hand‐sexed male fingerlings averaging 19±0.39 and 32±0.69 g were stocked in cages and open ponds at 150 fish cage^?1 and 2 fish m^?2 respectively. Fish were cultured for 114 days in five triplicate treatments. Cages were installed into ponds and caged fish were fed a 24% protein diet at 3% (T1) and 6% (T2) body weight day^?1 (BWD) without pond fertilization, and 6% BWD with pond fertilization (T3). The open water in the fourth treatment (T4) was not stocked but contained caged fish, which were fed 6% BWD for the first 57 days followed by 3% BWD for the remaining period. Ponds in the control (T5) had no cages and were neither fertilized nor open‐pond fish fed. Feeding rate and pond fertilization significantly (P<0.05) affected fish growth, profitability and water quality among treatments. Fish growth, feed utilization, fish yield, water quality and profits were significantly (P<0.05) better in T3 than the other treatments. It was concluded that fish production and economic returns were optimized at 6% BWD in fertilized ponds.     

Purification and characterization of cysteine proteinase inhibitors from crucian carp <Emphasis Type="Italic">Carassius auratus</Emphasis> eggs

Two cystatins (cst-I and cst-II) were purified from crucian carp eggs by acidification and subsequent ion exchange and molecular sieve chromatography. The molecular masses of cst-I and cst-II analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis were 11.9 and 14.4 kDa, respectively, under reducing conditions and 13.5 and 12.7 kDa, respectively, under non-reducing conditions. The cst-I and cst-II molecules were stable after 30 min of incubation at 60 and 50°C, respectively. There was no significant loss in the inhibitory activity of either cst in the pH range 4–11. These two cystatins were able to affect the proteolysis of papain, cathepsin L, and bromelain, but they were unable to inhibit cathepsin B and trypsin. The partial N-terminal amino acid sequences of both cst inhibitors were homologous and that of cst-I was recognized as NH₂-AGIPGGLVDADINDADVQ. This latter fragment shared 88.9% identity to common carp cystatin and 44.4–55.6% to cystatins of other aquatic animals. Based on these results, we conclude that the two cst inhibitors are members of family II cystatin.     

Influence of rubber trees on leaf-miner damage to coffee plants in an agroforestry system

The coffee leaf-miner (CLM) (Leucoptera coffeella Guérin-Mèneville; Lepidoptera: Lyonetiidae), the main pest of coffee plants, occurs widely throughout the Neotropics where it has a significant, negative economic and quantitative impact on coffee production. This study was conducted in a rubber tree/coffee plant interface that was influenced by the trees to a varying degrees depending on the location of the coffee plants, i.e. from beneath the rubber trees, extending through a range of distances from the edge of the tree plantation to end in a coffee monocrop field. The most severe damage inflicted on coffee plants by the CLM (number of mined leaves) from April, which marks the start of the water deficit period, until September 2003 was in the zone close to the rubber trees, whereas the damage inflicted on plants in the monocropped field was comparable to that on coffee plants grown directly beneath the rubber trees, which received about 25–40 % of the available irradiance (I_r—available irradiation at a certain position divided by the irradiation received in full sunlight, i.e. in the monocrop). From May until July damage caused by the CLM nearly doubled in each month. In midwinter (July), the damage decreased perceptibly from the tree edge toward the open field. From September onward, with the rising air temperatures CLM damage in the coffee monocrop started to increase. Based on these results, we conclude that coffee plants grown in the full sun incurred the most damage only at the end of winter, with warming air temperatures. Coffee plants grown in shadier locations (25–40 % I_r) were less damaged by the CLM, although a higher proportion of their leaves were mined. The rubber trees probably acted as a shelter during the cold autumn and winter seasons, leading to greater CLM damage over a distance outside the rubber tree plantation that was about equal to the height of the trees. Future studies should attempt to relate leaf hydric potential to pest attack in field conditions. More rigorous measurements of shade conditions could improve our understanding of the relationship of this factor to CLM attack.     

Birds like it Corky: the influence of habitat features and management of ‘montados’ in breeding bird communities

In the southwest part of the Iberian Peninsula the dominant land-use are the Portuguese montados and Spanish dehesas, parkland forested areas of anthropogenic origin dominated by cork oak. They form a wooded matrix with open areas, scattered woodlands and undisturbed patches of Mediterranean forest and scrublands. The montados are characterized by a rich bird community. We have focus our study in a multidisciplinary approach, evaluating how management and landscape patterns influence the bird community in order to identify potential threats to its conservation. The study was conducted in the Site of Community Importance of Serra de Monfurado were 70% of the area is cork and holm oaks. We used data from 120 10-min point counts. Using variation partitioning, we determined the independent and joint effects of Forest, Management and Habitat variables. The variation captured in bird assemblage was 65.06%. Most of the explained variation was related to the Habitat and Management variables. The explanatory variables that were high-lighted as important predictors were variables that reflected tree and shrub density and cork removal. Modelling for forest species through generalized linear models (GLM) emphasize that the management plays an important role in the species distribution. The most important variables selected in models reflected cattle grazing and cork removal. Our results point out that the type of management is crucial to maintain the equilibrium in bird community associated to montados. Farmland and forest species will benefit from areas with different tree densities, small patches of Mediterranean scrubs in the understory and correct livestock numbers.