Oleuropein-Specific-13-Glucosidase Activity Marks the Early Response of Olive Fruits （Olea europaea） to Mimed Insect Attack

Olive fruits are seriously deteriorated by pre and postharvest damage due to the attack of insects, such as Bactrocera olaea, which strongly alters the quality of olives. Defence response in olive fruits injured both by pathogens and by mechanical damages has been associated with the enzyme β-glucosidase, which specifically hydrolyses oleuropein, producing highly reactive aldehyde molecules. In situ detection of ~-glucosidase activity in olive fruit tissues following injury, which simulates Bactrocera oleae punctures, is reported. The assay was performed in two cultivars showing different degrees of susceptibilities to fly infestation. In both cultivars, the histochemical assay for β-glucosidase showed that within 20 min after the injury, a strong ~-glucosidase activity could be observed in the damaged tissues. Thereafter a progressive enzyme inactivation occurred starting from tissues around the boundary of the injury with decrease of the enzyme activity and stopped after 3 h. Whereas the mass of active cells reached a distance of （300±50） μm from the edge of the injury. Biochemical analyses showed that in extracts of the injured fruit, β-glucosidase activity rapidly increased within 20 min from injury, thereafter decreasing and reaching values comparable with those in intact fruits. Following puncture, the oleuropein contents did not change significantly in the high susceptibility cultivar, whereas it rapidly decreased in the cultivar showing low susceptibility. The results strongly suggest that olive fruits susceptible towards fly infestation could be related to the ability of the oleuropein-degrading-β-glucosidase to produce the highly reactive molecules in the damaged tissues. As a consequence of puncture, high level of peroxidase activity was detected. This feature also suggested that this enzyme could play a key role in the defence response against insect injuries.     

Oleuropein-Specific-β-Glucosidase Activity Marks the Early Response of Olive Fruits (Olea europaea) to Mimed Insect Attack

Olive fruits are seriously deteriorated by pre and postharvest damage due to the attack of insects, such as Bactrocera olaea, which strongly alters the quality of olives. Defence response in olive fruits injured both by pathogens and by mechanical damages has been associated with the enzyme β-glucosidase, which specifically hydrolyses oleuropein, producing highly reactive aldehyde molecules. In situ detection of β-glucosidase activity in olive fruit tissues following injury, which simulates Bactrocera oleae punctures, is reported. The assay was performed in two cultivars showing different degrees of susceptibilities to fly infestation. In both cultivars, the histochemical assay for β-glucosidase showed that within 20 min after the injury, a strong β-glucosidase activity could be observed in the damaged tissues. Thereafter a progressive enzyme inactivation occurred starting from tissues around the boundary of the injury with decrease of the enzyme activity and stopped after 3h. Whereas the mass of active celsreached a distance of (300±50)μm from the edge of the injury. Biochemical analyses showed that in extracts of the injured fruit, β-glucosidase activity rapidly increased within 20 min from injury, thereafter decreasing and reaching values comparable with those in intact fruits. Following puncture, the oleuropein contents did not change significantly in the high susceptibility cultivar, whereas it rapidly decreased in the cultivar showing low susceptibility. The results strongly suggest that olive fruits susceptible towards fly infestation could be related to the ability of the oleuropein-degrading-β-glucosidase to produce the highly reactive molecules in the damaged tissues. As a consequence of puncture, high level of peroxidase activity was detected. This feature also suggested that this enzyme could play a key role in the defence response against insect injuries.     

Effects of glandular trichomes on the development of Phytophthora infestans infection in potato (S. tuberosum)

S. berthaulltii Hawkes, a wild potato species, possesses two types of glandular trichomes on its leaf surface: types A and B wich confer resistance to insects. Preliminary results indicated that the presence of glandular trichomes also confers resistance to Phytophthora infestans, the causal agent of late blight. In this paper we report the evaluation of trichome characteristics (density and exudate activities) performed on 8 lines of potato, including the control varieties Desirée and Tropicana, 5 hybrid lines derived from intercrosses and backcrosses between S. tuberosum and S. berthaultii and one accession of S. berthaultii. Whole plants of the 8 genotypes were inoculated with a spore suspension of Phytophthora infestans in order to evaluate their resistance to late blight. The resultant disease incidence correlated negatively with type A trichome density and with polyphenol-oxidase (PPO) activity of type A trichome glands. The removal of type B trichomes exudate by leaf dipping in ethanol did not influence the disease development after artificial inoculation of detached leaves. Type B trichome exudate was extracted and the fraction corresponding to Fatty Acids Sucrose Esters (FASE) was recovered after Thin Layer Chromatography and tested on the fungal mycelium in vitro. The FASE exhibited a slight inhibitory effect on mycelium growth. The results of the experiments are discussed, and several hypotheses regarding the possible role of types A and B trichomes are formulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nitrogen and Phosphorus Levels in Sediments from Tropical Catatumbo River (Venezuela)

Nitrogen (N) and phosphorus (P) concentrations were determined in sediment samples along the bed of Catatumbo river in both Colombian and Venezuelan territories until the river outlet in Maracaibo lake. Total phosphorus was determined by digestion with HCl followed by analysis using the ascorbic acid method and total nitrogen was done using the standard microkjeldahl method plus nitrate-nitrite. Ammonium, orthophosphate and nitrate were determined using standard methods after extraction steps. The mean concentrations along the river bed were found in an interval of 0.035 and 1.492 mg g^-1 dry sed. for nitrogen and 0.027 and 1.039 mg g^-1 dry sed. for phosphorus at 95% confidence level. The mean molar ratio N/P in the river bed was 4.42 and 3.46 for river outlet zones in the lake, which indicates that nitrogen is the limiting nutrient. For comparison with previous results of lake sediments from sites near the river outlet it was concluded that Catatumbo river is a significant source of nutrients to the Maracaibo Lake system because sediment nutrients concentrations from Catatumbo river were higher than the ones in Maracaibo Lake. Statistic studies showed significant differences between countries, zones and similar behaviour in the river bed as related to the affluent rivers.     

Sustainable sediment management in coastal infrastructures through an innovative technology: preliminary results of the MARINAPLAN PLUS LIFE project

Journal of Soils and Sediments - The paper aims to show the preliminary monitoring and field test results of the innovative technology tested in the framework of the MARINAPLAN PLUS LIFE project...     

Achievements and future perspectives of embryo transfer technology in pigs

Commercial embryo transfer (ET) has unprecedented productive and economic implications for the pig sector. However, pig ET has been considered utopian for decades mainly because of the requirements of surgical techniques for embryo collection and embryo deposition into recipients, alongside challenges to preserve embryos. This situation has drastically changed in the last decade since the current technology allows non‐surgical ET and short‐ and long‐term embryo preservation. Here, we provide a brief review of the improvements in porcine ET achieved by our laboratory in the past 20 years. This review includes several aspects of non‐surgical ET technology and different issues affecting ET programmes and embryo preservation systems. The future perspectives of ET technology are also considered. We will refer only to embryos produced in vivo since they are the only type of embryos with possible short‐term use in pig production.     

Efficacy assessment of an intramammary treatment with a new recrystallized enrofloxacin vs ceftiofur and parenteral enrofloxacin in dairy cows with nonsevere clinical mastitis

A recrystallized form of enrofloxacin as dehydrate‐HCl (enro‐C) was assessed for bacteriological and clinical cure efficacies in Holstein‐Friesian cows affected of nonsevere clinical mastitis. Treatments were enro‐C_susp (n = 81), treated with a pharmaceutical suspension of enro‐C/quarter; group enro‐C_pd (n = 80) treated as above, but using enro‐C powder suspended in water; group CF (n = 65), treated with ceftiofur HCl/quarter; and group enro_R (n = 66), treated with standard enrofloxacin solution (5 mg/kg, intramuscular). Cows had a mean milk production of 31 L/day and were 2‐3 lactational periods old. Treatments were administered every 24 hr for 3 days. Groups treated with enro‐C exhibited statistically significant (p > .05) better clinical cure as compared to groups treated with CF or enro_R (95.06%, 96.25%, 67.79%, and 57.55%, for enro‐C_susp, enro‐C_pd, CF, and enro_R, respectively). In contrast, probability of bacteriological cure was not statistically different among treatments. Yet, the outstanding clinical and bacteriological cure rates obtained for enro‐C for nonsevere cases of mastitis is superior to previously reported data for parenteral enrofloxacin and other antibacterial‐intramammary treatments. Impact of using enro‐C on the rate and pattern of bacterial resistance, somatic cell counts and milk electric conductivity, must be studied. Also, the use of enro‐C for complicated cases of mastitis should be studied and milk withdrawal times must be accurately established.     

Antibiotic susceptibility of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolated from subclinical bovine mastitis cases and in vitro efficacy of bacteriophage

Staphylococcus aureus is an opportunistic pathogen that may cause severe infections in livestock, and represents the major cause of mastitis in dairy cows. Currently, instead of using antibiotics, new strategies are sought to reduce this clinical health problem. The aim of this study was to determine the efficacy of phage therapy to kill S. aureus strains obtained from farms located at the State of Guanajuato, México. Thirty-six S. aureus strains from cow milk with subclinical mastitis were isolated and identified, and the susceptibility to antibiotics and four phages also isolated in this work was tested. It was found that more of 90% of S. aureus isolates were not susceptible to six or more antibiotics, and 100% were resistant to penicillin, dicloxacillin, cefotaxime, ampicillin and cephalothin, and 81 and 77%, to tetracycline and cefuroxime, respectively. Fortunately, 100% of S. aureus isolates were susceptible to phages used in this work, which was detected as clear zones using specific phage. It was shown for the first time, that phages used in this study are active against pathogenic S. aureus and might be incorporated into the therapy as an important tool for the control of staphylococcal bovine mastitis, specially to antibiotic-resistant S. aureus strains isolated in farm located at the state of Guanajuato, México; and its use might be extended to other regions inside or outside the country.