Evaluation of different media and a BGM cell culture assay for isolation of Borrelia burgdorferi sensu lato from ticks and dogs

A co-culture assay for isolation of Borrelia burgdorferi sensu lato (sl.) from naturally infected ticks and dogs suspected of Lyme borreliosis (LB) was evaluated using buffalo-green-monkey (BGM) cells as the mammalian component. Four different media were tested for their ability to provide sufficient growth conditions for spirochetes and BGM cells. A total of 176 Ixodes ricinus ticks and 268 specimens from 98 dogs were used to compare cell-free culture with the BGM co-culture. A 1:1 mixture of Barbour-Stoenner-Kelly medium (BSK) and Eagle's minimum essential medium (EMEM) supported the growth of the two test strains, B. burgdorferi sensu stricto B31 and B. valaisiana VS116 to the same extent as BSK medium and the growth as well as the viability of BGM cells in this medium were the same as in EMEM. Using the 1:1 mixture of BSK and EMEM, borrelial growth measured in co-culture with BGM cells did not differ significantly from corresponding values obtained in cell-free cultures. In cell-free culture the isolation rate of B. burgdorferi sl. from ticks was significantly higher in BSK/EMEM 1:1 than in BSK medium (P < 0.01). Co-culture with BGM cells had no significant influence on the isolation rate of borreliae from ticks. However, a significant amount of isolates were obtained by one of the procedures only. Analysing canine specimens accordingly, spirochetes were grown from the blood of one dog after four weeks in BGM cell co-culture. The isolate was classified as B. afzelii by PCR-coupled restriction fragment length polymorphism analysis.     

Genetic similarity of natural populations and plantations of Pinus roxburghii Sarg. in Nepal

? Genetic structures of five population pairs each consisting of one natural population and one neighbouring plantation of Pinus roxburghii Sarg. in Nepal were assessed using nuclear (nSSRs) and chloroplast microsatellite loci (cpSSRs).

? The mean number of alleles at nSSRs loci in natural populations was 5.0 compared to 4.93 in plantations while the average observed heterozygosities were the same in both groups (H ₀ = 0.50). Most of the alleles were common to all the populations, indicating that the populations correspond to a single genetic entity. Similarly forty-seven haplotypes were observed in natural populations compared to fifty haplotypes in plantations. Mean haplotype diversities of natural populations (0.953) and plantations (0.955) were very similar. Genetic diversity of Pinus roxburghii was relatively high with low or no evidence of inbreeding while genetic differentiation among all populations was very low (about 1%).

? The very low differentiation among natural populations indicates efficient long-distance gene flow among populations resulting in homogeneous genetic structures at least at selectively neutral loci. Even though the harvest and production of seeds and seedlings was largely uncontrolled, genetic structures of most plantations show no signs of reduced variation, inbreeding or other negative effects compromising the adaptedness or adaptability of planted forests.

     

Mastication and prescribed fire impacts on fuels in a 25-year old ponderosa pine plantation, southern Sierra Nevada

Due to increases in tree density and hazardous fuel loading in Sierra Nevadan forests, land management is focusing on fuel reduction treatments to moderate the risk of catastrophic fires. Fuel treatments involving mechanical and prescribed fire methods can reduce surface as well as canopy fuel loads. Mastication is a mechanical method which shreds smaller trees and brush onto the surface fuel layer. Little data exist quantifying masticated fuel beds. Despite the paucity of data on masticated fuels, land managers desire fuel loading, potential fire behavior and fire effects such as tree mortality information for masticated areas. In this study we measured fuel characteristics before and after mastication and mastication plus prescribed burn treatments in a 25-year old ponderosa pine (Pinus ponderosa C. Lawson) plantation. In addition to surface fuel characteristics and tree data collection, bulk density samples were gathered for masticated material. Regressions were created predicting masticated fuel loading from masticated fuel bed depth. Total masticated fuel load prior to fire treatment ranged from 25.9 to 42.9 Mg ha⁻¹, and the bulk density of masticated fuel was 125 kg m⁻³. Mastication treatment alone showed increases in most surface fuel loadings and decreases in canopy fuel loads. Masticated treatment in conjunction with prescribed burning reduced both surface and canopy fuel loads. Detailed information on fuel structure in masticated areas will allow for better predictions of fire behavior and fire effects for fire in masticated fuel types. Understanding potential fire behavior and fire effects associated with masticated fuels will allow managers to make decisions on the possibility of mastication to create fuel breaks or enhance forest health.     

Activity Behavior of Horses Housed in Different Open Barn Systems

Housing conditions do not frequently meet the natural needs of horses. German and Swiss studies have shown that merely 16% of horses are kept in group housing systems, but even these vary greatly with different effects on horses’ behavior. The objective of the survey was to determine whether horses show increased activity in barns with various options for movements and functional elements. Free movement stables allow horses to live within a herd, to decide whether to be outside or inside the herd, and to enable them fresh air supply. Data from 25 horses kept in three different open barns (O1–O3) were compared with that from two active barns with functional elements (active barn 1 and active barn 2), for example, automatic feeder, with five horses each. Activity-Lying-Temperature-pedometers fastened to a rear leg of each horse were used to measure activity. Animal data were recorded throughout a test period of 10 days per barn. The results showed a median of 46 to 68 mean daily activity impulses per 10 minutes for horses in stables O1 to O3. In stables active barn 1 to active barn 2, this measure was exceeded considerably with medians of 77 and 151. The activity impulses in the stable systems were significantly different at an error level of α = 0.05. These results support previous studies and show that activity of horses depends on their opportunities for moving. The sectioning of a stable, placement of functional elements, and feeding frequency have considerable effects on the activity level of the horse. It is useful to integrate different functional areas in a group housing system.     

Somatic hybrids of vegetable brassicas as source for new resistances to fungal and virus diseases

Somatic hybrids were produced by PEG-induced symmetric and asymmetric protoplast fusions in order to transfer resistance to Alternaria brassicicola, A. brassicae, Phoma lingam, Plasmodiophora brassicae and Turnip mosaic virus (TuMV) into Brassica oleracea var. capitata (cv. ‘Toskama’) and botrytis (cv. ‘Korso’). As resistance donors, ten species belonging to several genera of the family Brassicaceae including wild relatives were used. Of 2,189 plants (somatic hybrids, partially in vitro cloned) tested, 1,616 (73.8%) were resistant against at least one of the pathogens, indicating that, mostly, a successful resistance transfer has taken place. Five hundred and twenty-two hybrids showed multiple resistances to two, three and, in a single case, to four pathogens. Irrespective of the donor parents used in the fusion, a broad variability in symptom manifestation ranging from 0 (without symptoms) to 9 (highly susceptible) could be observed. With regard to the Alternaria pathogens, Sinapis alba, B. nigra and B. juncea were the most effective resistance donors, whereas fusions with Raphanus sativus resulted in the most hybrids with resistance to clubroot and TuMV. As could be shown especially in asymmetric fusions with S. alba, Barbarea vulgaris and Hesperis matronalis, transferred resistance to a pathogen may not correspond with resistance exhibited by the donor parent. Some combinations in which both parents were highly susceptible, e.g. R. sativus (+) B. oleracea var. capitata, yielded hybrids that exhibited strong resistance, e.g. to A. brassicicola, revealing that a new type of resistance might be occurring. With regard to the Alternaria pathogens, resistance expression was very unstable. Many hybrids into which (also variable) resistance of some donors, such as B. vulgaris, S. alba and B. carinata, was transferred became as highly susceptible as those of which the fusion parents did not show any resistance reaction (e.g. R. sativus). For reliable characterization of the resistance response, hybrids should be subjected to several resistance tests during growth period of the host, at least until flowering.     

Search of environmental descriptors to explain the variability of the bacterial diversity from maize rhizospheres across a regional scale

The regional scale variability of the bacterial community inhabiting the rhizosphere was studied with soil collected from maize fields located in the Santo Domingo Valley (SDV; Baja California Sur, Mexico), a semi-arid agricultural ecosystem of approximately 200 km². The bacterial community structure was visualized by single-strand conformation polymorphism (SSCP) profiles of PCR-amplified partial 16S rRNA genes of directly extracted rhizosphere soil DNA. SSCP profiles of different SDV sites and an external field site in Germany were evaluated for their similarities and the contributing bacteria were characterized by DNA sequence analyses. SSCP profiles from each site were significantly different from the others, as revealed by permutation of pairwise similarities (P < 0.05). In comparison to the German site, SSCP profiles from SDV were more similar to each other despite contrasting soil salinity levels. Correspondence analysis revealed that among SDV sites, salinity levels, soil organic carbon and calcium (Ca²⁺) were most influential on the bacterial community structure. Depending on the phylogenetic group analyzed (Bacteria, Alphaproteobacteria, Pseudomonas), the importance of these soil variables varied. Interestingly, the East–West direction also revealed an effect, suggesting that future explorations of bacterial diversity patterns should also consider landscape topography in search of explaining patterns of bacterial diversity in soils.     

Effectiveness of a novel prostaglandin analog in postpartum mares and mares with early season anestrus

Contents Fifty thoroughbred mares, located at Central-Southern Brazil near Sao Paulo (13 anestrous maiden, 18 anestrous barren mares and 19 postpartum mares) were treated with one to three injections, 48 hrs apart, of 2 mg of the novel PGF analog K 11941*, during the early part of the breeding season. Early season anestrus in these problem mares was compounded by the adverse effects of a drought. Plasma progesterone determinations revealed unexpectedly elevated plasma progesterone levels in 61.5% of the maiden mares, in 77.8% of the barren mares and in 68.4% of the postpartum mares (20 to 25 days postpartum). Treatments with K 11941 initiated luteolysis, heat and ovulations in 91.4, 45.5 an 63.3% of these mares, respectively, but initiated also heat and ovulation in 13% and 33% of mares with baseline progesterone levels. Treatments initiated covert and overt cyclic functions in 17 and 12 of those animals which did not conceive immediately. Fifteen animals with recurrent anestrus received further treatment with K 11941. Ten mares cycling silently were force bred when palpations indicated ovulatory follicles, and 7 conceived. Of the 50 mares treated, 38 became pregnant (76%: maiden mares: 76.9%; barren mares: 77.8% and postpartum mares: 73.7%) with breeding indices of 1.9, 1.36 und 1.57, respectively; and 37 mares gave birth to a live foal (74%), At the same stud, 138 control mares had a foaling rate of 82.6%.