Tissue Culture Response of Ornamental and Medicinal Aesculus Species-A Review.

Species of the genus Aesculus are very attractive woody ornamentals. Their organs contain numerous health-promoting phytochemicals. The most valuable of them-aescin-is used in commercial preparations for the treatment of venous insufficiency. The industrial source of aescin is horse chestnut seeds because the zygotic embryos are the main site of its accumulation. Horse chestnut somatic and zygotic embryos contain similar amount of aescin, hence somatic embryos could be exploited as an alternative source of aescin. Somatic embryogenesis, androgenesis and de novo shoot organogenesis were successfully achieved in several Aesculus species, as well as secondary somatic embryogenesis and shoot organogenesis, which enables mass production of embryos and shoots. In addition, an efficient method for cryopreservation of embryogenic tissue was established, assuring constant availability of the plant material. The developed methods are suitable for clonal propagation of elite specimens selected as the best aescin producers, the most attractive ornamentals or plants resistant to pests and diseases. These methods are also useful for molecular breeding purposes. Thus, in this review, the medicinal uses and a comprehensive survey of in vitro propagation methods established for Aesculus species, as well as the feasibility of in vitro production of aescin, are presented and discussed.

Impact of Storage Temperature on Pollen Viability and Germinability of Four Serbian Autochthon Apple Cultivars.

Globalization has drastically reduced the number of autochthon apple cultivars in the Serbian market and most of them have nearly disappeared; however, some of these cultivars, such as Petrovaca, Budimka, Kolacara Pozna, and Kozara, have extraordinary quality, good pomological characteristics, and pest and disease resistance. The present study was conducted to develop a protocol for the storage of pollen for further use in the conservation and breeding of these cultivars. Viability and germination of the mature pollen were tested in vitro, at four storage temperatures (20, 4, -20, and -80°C), right after harvest or 1, 2, 3, 4, 5, and 6 months after storage. Differences in fresh pollen viability and germination between cultivars were statistically significant and ranged from 60 to 88% and 59 to 98%, respectively. Fresh pollen of cv. Budimka showed the highest viability and germination in comparison with other cultivars, especially cv. Kozara. Pollen viability and germination decreased over the storage period, and it was the lowest after 6 months of storage at room temperature in all tested cultivars. Storage at 4°C prolonged the pollen viability and germinability of 1-5 fold, depending on the cultivar and treatment duration; however, the pollen longevity of all cultivars was significantly extended when stored at -20 or -80°C. After 6 months, pollen of cv. Budimka stored at -20 and -80°C showed 14-15 fold higher germination rates in relation to pollen storage at room temperature for the same period. The results of the present study suggest that the pollen of these apple cultivars could be efficiently maintained at -20°C and could be further used for breeding purposes, e.g., for crossings between cultivars that flower at different times of the year.

Variability in somatic embryo-forming capacity of spinach.

High variability in somatic embryo (SE)-forming capacity has previously been observed in several spinach cultivars. Such variability frequently accounted for more variation in embryogenic response of the explants than the factor being investigated. Hence, the variability in embryogenic capacity was examined in the present study at both the population and the single-seedling level, using seeds of spinach cultivar Matador obtained from nine European seed companies. Seed population obtained from Slovenia (Sl) was superior to others, with the highest regeneration frequency (100%) and the highest mean SE number (14.4). A total of 82% of these seedlings had 80-100% of regenerating explants, while in populations with intermediate embryogenic capacity approximately 40% of seedlings had 20-60% of regenerating explants. The explants from the majority of seedlings (52-100%) in the least responsive populations were irresponsive. Furthermore, the explants from Sl seedlings regenerated from 10-20 (43.5%) up to > 20 (27.6%) SEs on average, while the explants from the majority of seedlings belonging to other populations regenerated 1-10 SEs. The present study strongly indicates that the variability of plant material must not be overlooked, because choosing more responsive individuals for one treatment and less responsive ones for another may lead to misinterpretation of the data.

The Effect of a Moderate Exercise Program on Serum Markers of Bone Metabolism in Dogs.

The beneficial effect of physical activity on the musculoskeletal health in dogs is well recognized, but the level of intensity, duration, and frequency of exercise is not fully described. Measurement of serum markers of bone metabolism (bone alkaline phosphatase and osteocalcin as bone formation markers and C-terminal telopeptide as bone resorption marker) during four months of organized moderate-intensity physical training in Labrador retriever and Golden retriever dogs aged between 11.7-24.4 months, showed variations of bone metabolism. Dogs were included in treadmill running sessions for 25 min, three times per week. Blood samples were taken at the beginning of the program (baseline), after two months (mid-term) and at the end of the study after four months. The values of bone alkaline phosphatase and osteocalcin significantly decreased following two months of exercise program. Bone alkaline phosphatase increased by the end of four-month training cycle, but did not reach baseline value. Osteocalcin levels continued to decrease towards the end of the study. C-terminal telopeptide concentrations did not significantly change throughout the study duration. The results of this study show that aerobic exercise of moderate-intensity caused an initial decrease in bone formation followed by an increase of bone alkaline phosphatase and a further decrease of osteocalcin concentration. The response of two formation markers can be explained by the different stage of osteoblast activity that they express. In summary, moderate exercise resulted in no change in bone resorption, and a mild bone formation in young developing dogs.

Heart rate deflection point during incremental test in competitive agility border collies.

The aim of this study was to determine a heart rate deflection point (HRdp) in competitive agility dogs. Fourteen healthy border collies underwent progressive incremental treadmill exercise--modified Conconi test protocol. Heart rate was continuously recorded, and the HRdp was estimated using two methods: subjective and computer aided regression. Maximal heart rate (HRmax), achieved running speed at the anaerobic threshold and at the end of test were also determined. Statistical analysis showed a very high positive correlation between HRdp determined by two methods. The mean HRdp in this research corresponded to 80% of HRmax. The wide range of individual HRdp (162-229 BPM) indicates the need of an individual approach in assessing physiological parameters. To the authors' knowledge, this is the first application of Conconi modified test for estimating anaerobic threshold in agility dogs since the standard for dogs is yet undetermined. Future studies need to detect the most appropriate and reliable technique for its determination as well as its applicability in programming of the optimal training intensity.

Animal assisted therapy and perception of loneliness in geriatric nursing home residents.

Use of animals for therapeutic purposes, animal assisted therapy or AAT is a method for improving quality of life for long-term inpatients. The object of this paper was to evaluate dog companionship as a form of AAT and its effects on perception of loneliness in geriatric nursing home residents. The participants were involved in a six-month program of dog companionship three times weekly for 90 minutes. There were 21 residents included in the program, with a mean age of 80 years. Loneliness was measured by the short version of the UCLA Scale of loneliness. Comparison of test results before and after participation in the program showed that dog companionship reduces the perception of loneliness.

The influence of sex on biochemical markers of bone turnover in dogs.

Biochemical markers of bone turnover have been shown to be useful as inexpensive and noninvasive tools for monitoring skeletal health. The reference range for bone markers in dogs has been set by different age groups. However, other sources of biological variations were not fully investigated in dogs. To explore whether sex influences the interpretation of bone marker data we examined serum bone markers in 33 male and 25 female dogs. The bone markers selected for this study were: bone alkaline phosphatase (BALP) and osteocalcin (OC) as indicators of bone formation, and C-terminal telopeptide (CTx) of type I collagen as marker of bone resorption. All concentrations of bone markers were lower, but still within the reference range reported for dogs. We found statistically significant differences of the median OC and CTx serum concentrations between males and females. The results of this study suggest that there are sex differences in biochemical markers of bone turnover in dogs which should be considered in interpretation of bone marker data.

Serum platelet-activating factor acetylhydrolase and paraoxonase-1 activity in horses infected with Leptospira spp.

The aim of the study was to investigate the effect of infection with Leptospira spp. in horses on activities of platelet-activating factor acetylhydrolase (PAF-AH) and paraoxonase-1 (PON1) considering the anti-inflammatory/anti-oxidative properties of both enzymes. A total of 63 sport and leisure horses' sera were chosen among those routinely serologically examined on leptospirosis during the year 2009 in Croatia. Sera were divided into three groups according to the estimated level of antibody titre against Leptospira spp; group 1: sera serologically negative to leptospirosis (antibody titre<50) (n=21); group 2: sera with residual or anamnestic antibody titre (50-200) (n=23); group 3: sera with high antibody titre (≥1600) indicating recent Leptospira infection (n=19). Serum PAF-AH and PON1 activity was not significantly different between investigated groups (p>0.05). There were no statistical differences in lipid status parameters (p>0.05) among study groups as well. However, significant positive correlations (p<0.05) of PAF-AH and PON1 with total cholesterol and HDL-C were found. Results may indicate low levels of systemic inflammatory response and oxidative stress in horses with subclinical leptospirosis. Further studies with clinically manifested disease are needed to elucidate the potential role of PAF-AH and PON1 as prognostic markers of the leptospirosis outcome.