Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit

The ellagic acid, total phenolic, and vitamin C contents in four raspberry cultivars (Heritage, Autumn Bliss, Rubi, and Zeva) grown in Spain were detected and quantified by HPLC in fresh, just frozen, and stored fruits at -20 degrees C for a one year period. Ellagic acid [207-244 mg kg(-)(1) of fresh weight (fw)], total phenolic (137-1776 mg kg(-)(1) of fw), and vitamin C (221-312 mg kg(-)(1) of fw) contents in raw material were higher in the late cultivars Zeva and Rubi than in the early cultivars Autumn Bliss and Heritage. The freezing process slightly affected the values of extracted ellagic acid, total phenolic, and vitamin C content. At the end of long-term frozen storage (12 months), no significant change of total phenolic content extracted was observed, but significant decreases of 14-21% in ellagic acid and of 33-55% in vitamin C were quantified. Free radical scavenging capacity measured as antiradical efficiency (AE) depends on the seasonal period of harvest. Late cultivars, Rubi (6.1 x 10(-)(4)) and Zeva (10.17 x 10(-)(4)), showed higher AE than early cultivars, Heritage (4.02 x 10(-)(4)) and Autumn Bliss (4.36 x 10(-)(4)). The freezing process produced a decrease of AE values in the four cultivars ranging between 4 and 26%. During the frozen storage, the AE values reached after the freezing process remained unchanged.     

Partial characterization of polyphenol oxidase activity in raspberry fruits.

A partial characterization of polyphenol oxidase (PPO) activity in raspberry fruits is described. Two early cultivars harvested in May/June (Heritage and Autumm Bliss) and two late cultivars harvested in October-November (Ceva and Rubi) were analyzed for PPO activity. Stable and highly active PPO extracts were obtained using insoluble poly(vinylpyrrolidone) (PVP) and Triton X-100 in sodium phosphate, pH 7.0 buffer. Polyacrylamide gel electrophoresis of raspberry extracts under nondenaturing conditions resolved in one band (R(f)()(1) = 0.25). Raspberry PPO activity has pH optima of 8.0 and 5.5, both with catechol (0.1 M). Maximum activity was with D-catechin (catecholase activity), followed by p-coumaric acid (cresolase activity). Heritage raspberry also showed PPO activity toward 4-methylcatechol. Ceva and Autumm Bliss raspberries showed the higher PPO activity using catechol as substrate.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Effect of different methods of administration of ivermectin on its efficacy against the shedding of gastrointestinal nematode eggs by gazelles

Seven different methods of administering ivermectin to gazelles were compared: subcutaneous injection, direct oral administration, administration in individual feeds, administration in a herd feed, direct oral administration of a second ivermectin formulation, administration in individual water supplies, and administration in the herd's water supply. The first five treatments were effective, as monitored by faecal egg count reduction tests, and administration in individual feeds or in a herd feed avoided the need to capture the animals, with the attendant risk of mortality. Of the factors associated with the recipients (species, sex, age and inbreeding coefficient) age was the only significant factor for the efficacy of the treatment. Oral or subcutaneous, individual or collective, and direct or indirect administrations were equally satisfactory for the treatment of all the parasite groups studied. Only when parasitic problems were due to Nematodirus species did direct administration to individual animals appear to be preferable.     

Experimental infection with Tritrichomonas suis in heifers.

Nine heifers were intravaginally challenged with 9.3x10(6) Tritrichomonas suis reference strains. Vaginal mucus and serum samples were collected weekly 4 weeks post-inoculation. Vaginal mucus was cultured for T. suis and sera was tested by ELISA against whole cell antigens for T. suis and Tritrichomonas foetus. All vaginal mucus cultures were T. suis-negative during the experiment. ELISA values for both antigens were similar and differences were not significant (P>0.05). Positive control serum samples from one heifer vaccinated against T. foetus showed anti-T. suis ELISA values. We concluded that T. suis intravaginal inoculation induced a low level of serum immune response in heifers measured by ELISA and both protozoa probably share a common antigen. However, under the experimental conditions of this trial, colonization of the heifers' genital tract was not possible in any of the nine animals.     

Determination and persistence of imazalil in post-harvest-treated apples during cold storage

A method to determine imazalil (allyl 1 -(2, 4-dichlorophenyl)-2-imidazol- 1-ylethyl ether) on apples using high performance liquid chromatography is described. After harvest, fruits were immersed in an aqueous suspension (1 g litre-1) and cold stored (0-2°C) at 85-90% r.h. Samples were taken monthly and imazalil was determined in the peel, outer pulp, inner pulp and on whole fruit. Residues were 3.8-4.9 mg imazalil kg-1 and decreased during storage. Most imazalil was found on the peel and amounts in the pulp decreased toward the core.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.