A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

Pilot study of the ultrasonographic examination of the intact and transected medial glenohumeral ligament in dogs

Medial glenohumeral ligament injury is commonly reported during medial shoulder joint instability in dogs. Arthroscopy is considered the gold standard procedure, but it is invasive and requires distension of the joint. Ultrasonographic examination of the medial glenohumeral ligament has been studied as a possible, less invasive alternative to arthroscopy however it has not been considered a useful method of assessment due to the interference of the probe with the pectoral muscles. The aims of this prospective analytical randomized pilot study were to develop a standardized ultrasound protocol for visualizing the canine medial glenohumeral ligament and to compare goniometry and ultrasound findings in cadaver dogs with versus without transection of the medial glenohumeral ligament. Nine adult Beagle cadavers (18 shoulders) were used. The first six shoulders were used in a preliminary study to describe an ultrasound technique to identify the medial glenohumeral ligament. Arthroscopy was performed on the remaining 12 shoulders, with six randomly selected medial glenohumeral ligaments from these shoulders, transected during the procedure. Ultrasound examination was performed after each arthroscopic procedure by an ultrasonographer blinded to the patient group. Four medial glenohumeral ligaments (67%) were correctly identified during the preliminary study. Ultrasonographic examination failed to diagnose the transection of all six medial glenohumeral ligaments in the second part of the study. No difference was observed in the ligament thickness between the dogs with and without a transected medial glenohumeral ligament. Dogs with a transected medial glenohumeral ligament had a wider articular space compared to dogs without a transected ligament (P < 0.001), and an articular space wider than 8.2 mm was discriminatory of a transected medial glenohumeral ligament in all the shoulders. In conclusion, the medial glenohumeral ligament could be identified with a medial ultrasonographic approach of the shoulder and a wider articular space can be a sign of a medial shoulder joint instability. Further studies are needed to confirm these preliminary findings in living dogs, with and without shoulder instability.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.     

Processing tomato quality as affected by irrigation scheduling

In Southern Italy the paucity of water is a very important problem and due to the fundamental importance of the water for the processing tomato, in this study the influence of irrigation on processing tomato quality was investigated in the years 2002 and 2003. A hybrid processing tomato (Lycopersicon esculentum L. Mill cv. Ability) was treated with different water regimes obtained combining amount of water and irrigation interval. The effects of the trials were evaluated taking into account the physical and chemical characteristics of the fruits, as well as the content in antioxidants moieties. Furthermore, the relationship among all the quality parameters and the seasonal amount of irrigation water was evaluated.The results, comparing different water regimes obtained combining amount of water and irrigation interval, gave useful indications on the possibility to improve nutritional tomato quality by reducing irrigation water during tomato cropping. In particular, relatively to the Mediterranean areas of tomato cultivation and to the texture of the soil considered in this study, the best compromise between quality and quantity of the processing tomato fruit was achieved both with the extension of the irrigation interval (L40 or L60) and with the limitation of irrigation volume for the second part of the tomato crop cycle (thesis 100-50). Extending the irrigation interval and limiting irrigation volume for the second part of the tomato crop cycle, appeared to be the best management strategy to optimize the yield and nutritional quality of processing tomato.     

Insights into the Effect of a Natural Arsenate Dose on Growth, Nodulation and Redox Metabolism of Soybean Plants

<正>Dear Editor,Arsenic(As) is a harmful metalloid that occurs in soil and water; its concentration varies considerably among geographic regions, with groundwater being the principal source of human contamination(Smedley and Kinniburgh, 2002). Besides the direct contamination effect of drinking water that contains high As concentration, human poisoning may also occur after inges-     

Nutritive and antioxidative potential of fresh and stored pomegranate industrial byproduct as a novel beef cattle feed

Pomegranate peel is a nutritive-rich byproduct whose amounts are extensively growing due to the exponential increase in the production of pomegranate juice and "ready to eat" arils. Pomegranate peel is a rich source for antioxidants and thus may serve in the prevention of cattle diseases and in the improvement of beef products, making it an attractive component in beef cattle diets. The present study aims to evaluate the effect of commonly used storage practices on the nutritive and antioxidative properties of pomegranate peel. In general, storage conditions preserved most antioxidant capacity. Ensiling ambivalently affected the nutritive values of the peel and promoted increased levels of antioxidative components. In addition to polyphenols, nonphenolic components, such as alpha- and gamma-tocopherols, contributed to the total antioxidative capacity, and several minerals found in the peel added to its nutritional value. Dietary supplementation with fresh peels promoted significant increases in feed intake and alpha-tocopherol concentration in the plasma, with positive tendency toward increased weight gain of bull calves. All in all, the nutritive value and the antioxidant capacity of pomegranate peel turn it into a favorable health-promoting constituent of feedlot beef cattle diet.     

Effects of Pre-Stratification Storage Conditions on Black Walnut Seed Post-Stratification Germination Capacity

The objective of this work was to quantify changes in the germination capacity of Juglans nigra seeds following storage and then artificial stratification. Seeds were stored at 5 and –20 °C under 5% and 14% internal moisture content (IMC) for 0, 8, 10, and 12 months under conditions with oxygen or under vacuum. Germination percentage (GP) was affected by storage temperature, IMC, and storage period. Seeds stored with 5% IMC succeeded in germinating regardless of the temperature. Seeds stored with 14% IMC did not germinate (–20 °C) or germinated during storage (5 °C). The GP was reduced with the time of storage in seeds at –20 °C and 5% IMC and this effect was avoided in storage under vacuum. In conclusion, the germination capacity of black walnut seeds did not change for up to 12 months at low above-zero temperature and low IMC (5 °C, 5% IMC), being the most cost-effective alternative for long-term storage of black walnut seeds in commercial nurseries. Vacuum storage avoided the decrease of around 50% in average germination capacity of seeds stored for a year at –20 °C.     

Parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> and role of the gelatinous matrix

Trichoderma (T. asperellum-203, 44 and GH11; T. atroviride-IMI 206040 and T. harzianum-248) parasitism on Meloidogyne javanica life stages was examined in vitro. Conidium attachment and parasitism differed beween the fungi. Egg masses, their derived eggs and second-stage juveniles (J2) were parasitized by Trichoderma asperellum-203, 44, and T. atroviride following conidium attachment. Trichoderma asperellum-GH11 attached to the nematodes but exhibited reduced penetration, whereas growth of T. harzianum-248 attached to egg masses was inhibited. Only a few conidia of the different fungi were attached to eggs and J2s without gelatinous matrix; the eggs were penetrated and parasitized by few hyphae, while J2s were rarely parasitized by the fungi. The gelatinous matrix specifically induced J2 immobilization by T. asperellum-203, 44 and T. atroviride metabolites that immobilized the J2s. A constitutive-GFP-expressing T. asperellum-203 construct was used to visualize fungal penetration of the nematodes. Scanning electron microscopy revealed the formation of coiling and appressorium-like structures upon attachment and parasitism by T. asperellum-203 and T. atroviride. Gelatinous matrix agglutinated T. asperellum-203 and T. atroviride conidia, a process that was Ca²⁺-dependent. Conidium agglutination was inhibited by carbohydrates, including fucose, as was conidium attachment to the nematodes. All but T. harzianum could grow on the gelatinous matrix, which enhanced conidium germination. A biomimetic system based on gelatinous-matrix-coated nylon fibers demonstrated the role of the matrix in parasitism: T. asperellum-203 and T. atroviride conidia attached specifically to the gelatinous-matrix-coated fibers and parasitic growth patterns, such as coiling, branching and appressoria-like structures, were induced in both fungi, similarly to those observed during nematode parasitism. All Trichoderma isolates exhibited nematode biocontrol activity in pot experiments with tomato plants. Parasitic interactions were demonstrated in planta: females and egg masses dissected from tomato roots grown in T. asperellum-203-treated soil were examined and found to be parasitized by the fungus. This study demonstrates biocontrol activities of Trichoderma isolates and their parasitic capabilities on M. javanica, elucidating the importance of the gelatinous matrix in the fungal parasitism.