Rapid detection and identification of ‘Candidatus Phytoplasma pini’‐related strains based on genomic markers present in 16S rRNA and tuf genes

In order to devise a method for rapid detection of ‘Candidatus (Ca.) Phytoplasma pini’ and for distinguishing it rapidly from other phytoplasmas, we carried out preliminary sequencing of Lithuanian ‘Ca. Phytoplasma pini’ strain PineBL2 using Illumina (NGS) technology and targeted sequencing employing universal phytoplasma primers. We focused on two resulting chromosomal segments that contained a 16S rRNA gene and a translation elongation factor EF‐TU gene (tuf), respectively. Based on alignments of the ‘Ca. Phytoplasma pini’ gene sequences with the corresponding sequences of other phytoplasmas, we designed new primer pairs for PCR‐based detection of ‘Ca. Phytoplasma pini’. Because ‘Ca. Phytoplasma pini’ strains are expected to reside in the pine phloem in a very low titre, one might expect that they could be detected only by nested PCR. By contrast, the primers and PCR protocols designed in the current work enabled rapid direct PCR detection and identification of ‘Ca. Phytoplasma pini ’ by amplifying a 484 bp 16S rDNA segment and a 513 bp tuf gene fragment that contain regions unique to this phytoplasma .     

Histomorphometry and μCT scan analysis of osteoporosis in spayed female dogs

Background:Both humans and small animals suffer from similar metabolic and structural diseases that impact the musculoskeletal system; however, instead of studying animal disease in its own right, animals are more often used as models for research into various human ailments, such as osteoporosis. There are few studies indicating that animals may suffer from osteoporosis, which raises the question of why small animals, which we believe to be equally susceptible, receive so little attention. With this research, we hope to draw the attention of researchers to the fact that the examination of animals for this disease is just as important as the examination of humans; human osteoporosis research receives a great deal of attention, while animals and their health are neglected.Aims:We aimed to analyze the bone volume fraction (BV/TV) and thickness of first (L1) and second (L2) lumbar vertebrae samples from five cadavers using histomorphometric analysis. In addition, we aimed to investigate one cadaver using microcomputed tomography (μCT) imaging.Methods:The L1 and L2 vertebrae from five dog carcasses were used to evaluate the BV/TV and the trabecular thicknesses. We used precise sampling criteria, and also developed a methodological approach to the study of the vertebrae. Using semi-automated methods, we performed histomorphometric analysis and μCT data analysis.Results:We used five dog cadavers in this research. During the histomorphometry study, we observed that the lowest L1 BV/TV ratio was 7.88% and the highest was 23.08%. The L2 vertebrae BV/TV ranged from 11.58% to 23.7%. The L1 and L2 lumbar trabeculae thicknesses were also measured. L1’s smallest trabecula was 17.34 microns and its largest was 31.88. The L2 vertebrae trabecula thickness was 18.76–30.75 microns. BV/TV and trabecular thickness were positively correlated (and vice versa). The two-tailed p value was less than 0.00001. This difference is statistically significant. After μCT analysis, we discovered regions in the vertebral body with low porosity; these cavities are usually filled with connective tissue. The bone tissue in these areas is more vulnerable, meaning fracture risk has increased.Conclusions:Animals should not just be considered as models for osteoporosis in humans, but also as potential patients. A single test, such as histomorphometry, may not be sufficient; more advanced technology, such as μCT, is required, since it reveals the pores that make the vertebral column more brittle and susceptible to fracture.     

secA gene suitability for fast and easy identification of Phytoplasmas by RFLP analysis

European Journal of Plant Pathology - In this study we investigated the secA gene as possible marker to supplement the classification scheme based on 16S rRNA sequences. We cloned and sequenced...     

Dynamic response of tree growth to changing environmental pollution

Long-term investigations of Scots pine (Pinus sylvestris L.) growth were carried out in the vicinity of one of the biggest air pollution sources in Lithuania—mineral fertilizers plant “Achema.” It is detected that initial stages of the dynamic tree response to the external stress factors in the polluted environment with an increased quantity of nitrogen compounds have started with a stimulation stage, followed by a gradual transition to a depression of growth. The recovery of the damaged stands took place along with the reduced environmental pollution, and the overdraft of the “normal” annual increment was a characteristic feature of all the investigated stands. This phase is still continuing for the most damaged stands. A very high individual variability of the tree growth response to the environmental impact is a characteristic feature of the damaged Scots pine stands. The homeostatic mechanisms of the survived trees stipulated reaching approximately the same or even higher growth rate as prior to the depression period, and the tree growth rate before the depression period can serve as the most powerful predictor of tree growth recovery capacity under the reduced environmental pollution. Crown defoliation is the next most important predictor of individual tree recovery capacity. Lower stand density and lower competition cause higher recovery capacity of damaged trees. The conclusion is made that a reduction in the environmental pollution on the local and regional scale and especially a decrease in emissions and deposition of sulfur and nitrogen compounds caused the recovery of damaged forests. These trends should be taken into account while analyzing and modeling forest dynamics. Interaction of environmental pollution and climatic factors is very important for the response of tree growth to the environmental stressors.     

Asymptomatic weeds are frequently colonised by pathogenic species of Fusarium in cereal‐based crop rotations

Several Fusarium species cause harmful cereal diseases, such as fusarium head blight and crown rot, which, during pathogenesis, may result in significant grain yield and quality losses. Several species of agricultural weed are believed to be alternative and reservoir hosts for Fusarium spp.; however, studies have not comprehensively evaluated those weed species in cropping systems that may harbour these fungi. The objective of this study was to determine weed species in cereal‐based crop rotations that are asymptomatically colonised by Fusarium spp. We sampled all species of weed present in fields that were managed under six different crop sequences in 2015 and 2016. The study yielded 2326 single‐spore isolates of Fusarium spp. derived from various organs of asymptomatic weeds. Isolates were identified morphologically and then confirmed using PCR with species‐specific primers and/or sequencing of tef1α gene fragments. Isolates of nine Fusarium spp. were obtained from 689 of the 744 individuals collected that represented 56 weed species. Each weed species harboured at least one species of Fusarium, and >80% were colonised by 3–9 Fusarium spp. In total, we identified 27 dicotyledonous weed species that were previously undocumented as Fusarium hosts and 251 new weed × Fusarium species combinations were revealed. Consequently, there is a greater risk of negative Fusarium impacts on cereal crops than was previously thought. We suggest effective weed management and inversion soil tillage may help mitigate these impacts.     

The seasonal variability of air pollution effects on pine conditions under changing climates

The multiple-stressor effects of air pollution, nutrient and water availability are the key issues of present forest ecosystem research. However, too little is known about the seasonal effect of pollutants on tree crown defoliation and their interaction with changes in climate. Therefore, data on seasonal variation in air pollution, including surface ozone, deposition of acidifying compounds and meteorological conditions, were tested against pine defoliation to identify the periods when the effect of the considered contaminants is most pronounced. The findings of the study revealed that a higher level of air concentrations of acidifying pollutants and their deposition was observed during the dormant period, with the exception of only ΣNH4⁺ air concentrations and their monthly deposition. An increase in precipitation over the vegetation period and mean monthly temperature from September to December, as well as a decrease in temperature and precipitation over the remaining months of the dormant period represented the climate change condition over the 14-year period in the observed region. Detected changes in the considered parameters during the dormant period were found to be most significant to changes in pine crown defoliation. Therefore, we concluded that climate changes, if they occur by the detected scenarios, should mitigate the negative effect of air pollutants and acid deposition on pine crown condition.     

Comparative Analysis of the Major Metabolites of Ukrainian Saffron Samples by HPLC

Plant Foods for Human Nutrition - Crocus sativus L. is one of the most valuable crops, which stigmas are used as the food supplement and herbal medicine. In Ukraine, the cultivation of C. sativus...     

Prevalence and epidemiological features of bovine viral diarrhoea virus infection in Lithuania

The objectives of the present work were to estimate the level of bovine viral diarrhoea virus (BVDV) infection in cattle herds at the different Lithuanian districts and to determine factors influencing the course of BVDV infection. The studies were explored in 147 intensive dairy cattle breeding herds from 27 different Lithuanian regions in 1997-2001. BVDV infection was diagnosed in all investigated regions. The existing variations in the structure of cattle population determined different distribution patterns of BVDV infection. The number of seropositive animals ranged from 11.9 to 100%. It must be pointed out that 29.9% of the herds were not infected with BVDV and in 32.7% of the herds from 70 to 100% of cattle were seropositive to BVDV. A positive correlation between the number of seropositive cattle, and the size of herds and age of animals was determined. Sex of animal had no influence on the prevalence of BVDV. It was estimated that the annual incidence risk of infection with BVDV decreases with the animal age.