Phytoplasmas infecting sour cherry and lilac represent two distinct lineages having close evolutionary affinities with clover phyllody phytoplasma

Phytoplasmas infecting sour cherry and lilac in Lithuania were found to represent two lineages related to clover phyllody phytoplasma (CPh), a subgroup 16SrI-(R/S)C (formerly 16SrI-C) strain exhibiting rRNA interoperon sequence heterogeneity. 16S rDNAs amplified from the cherry bunchy leaf (ChBL) and lilac little leaf (LcLL) phytoplasmas were identical or nearly identical to those of operon rrnA and operon rrnB, respectively, of CPh. There was no evidence of 16S rRNA interoperon sequence heterogeneity in either LcLL or ChBL phytoplasma. Based on collective RFLP patterns of 16S rDNA, ChBL was classified in subgroup 16SrI-R, and LcLL was classified in new subgroup 16SrI-S. The ribosomal protein (rp) gene sequences from LcLL phytoplasma were identical to those of CPh, and strain LcLL was classified in rp subgroup rpI-C. By contrast, rp gene sequences from ChBL phytoplasma differed from those of subgroup rpI-C; based on RFLP patterns of rp gene sequences, ChBL was classified in new rp subgroup rpI-O. Single nucleotide polymorphisms (SNPs), designated here by a new SNP convention, marked members of rp subgroup rpI-C, and distinguished LcLL and CPh from ChBL and other non-rpI-C phytoplasmas in group 16SrI. The results raise questions concerning phytoplasma biodiversity assessment based on rRNA genes alone and encourage the supplemental use of a single copy gene in phytoplasma identification and classification, while drawing attention to a possible role of horizontal gene transfer in the evolutionary history of these lineages.     

Biochemical Characterization of Cassiopea andromeda (Forsskål, 1775), Another Red Sea Jellyfish in the Western Mediterranean Sea

Increasing frequency of native jellyfish proliferations and massive appearance of non-indigenous jellyfish species recently concur to impact Mediterranean coastal ecosystems and human activities at sea. Nonetheless, jellyfish biomass may represent an exploitable novel resource to coastal communities, with reference to its potential use in the pharmaceutical, nutritional, and nutraceutical Blue Growth sectors. The zooxanthellate jellyfish Cassiopea andromeda, Forsskål, 1775 (Cnidaria, Rhizostomeae) entered the Levant Sea through the Suez Canal and spread towards the Western Mediterranean to reach Malta, Tunisia, and recently also the Italian coasts. Here we report on the biochemical characterization and antioxidant activity of C. andromeda specimens with a discussion on their relative biological activities. The biochemical characterization of the aqueous (PBS) and hydroalcoholic (80% ethanol) soluble components of C. andromeda were performed for whole jellyfish, as well as separately for umbrella and oral arms. The insoluble components were hydrolyzed by sequential enzymatic digestion with pepsin and collagenase. The composition and antioxidant activity of the insoluble and enzymatically digestible fractions were not affected by the pre-extraction types, resulting into collagen- and non-collagen-derived peptides with antioxidant activity. Both soluble compounds and hydrolyzed fractions were characterized for the content of proteins, phenolic compounds, and lipids. The presence of compounds coming from the endosymbiont zooxanthellae was also detected. The notable yield and the considerable antioxidant activity detected make this species worthy of further study for its potential biotechnological sustainable exploitation.     

Structure and hydraulic properties of the boreal clay soil under differently managed buffer zones

Vegetated buffer zones (BZs) between arable fields and bodies of water are commonly established to reduce erosion and run‐off of particle‐bound nutrients. Functioning of a BZ depends on soil structure, as it is important for water infiltration. Therefore, it is vital to understand how varying management practices affect soils of BZs. We studied the structural and hydraulic properties of three differently managed BZs established in a boreal Vertic Stagnic Cambisol (clay, 51%). The three management practices for vegetation were as follows: natural with no treatment, harvested yearly and grazed by cattle. We used bulk density and macroporosity, together with a pore geometry index (air permeability per unit air‐filled porosity), to describe the soil structural properties. Hydraulic properties were measured at different length scales by means of an aggregate sorptivity test, saturated hydraulic conductivity of the core samples and field‐saturated hydraulic conductivity. Vegetation management markedly affected the physical properties in the top 5 cm of the soil. Properties were least favourable for infiltration at the grazed site, with the greatest bulk density, least macroporosity and hydraulic conductivity or greatest pore tortuosity. In general, spatial variation in zones with restricted and good hydraulic conductivity together with reduced aggregate sorptivity in the deeper horizons made the soil prone to preferential flow when initially dry. Prolonged wetness, on the other hand, reduced saturated hydraulic conductivity significantly, resulting in surface run‐off. Harvesting was considered the best management practice due to its inherent capacity for reducing the soil nutrient content and because it has minor implications for soil physical properties.     

Phosphorus exchange properties of European soils and sediments derived from them

The sorption and desorption of phosphorus (P) from eroding soil particles in land runoff are important processes contributing to agriculturally‐driven eutrophication. We investigated the P‐exchange properties and related chemical characteristics of contrasting European agricultural soils and sediment material eroded from them under indoor (small‐scale) and outdoor (larger‐scale) rainfall simulations. Quantity‐intensity (Q/I) relationships revealed large variation in equilibrium P concentrations at zero net P sorption (EPC₀) (0–10.3 mg l⁻¹) and instantly labile P (−Q₀, the amount of P to be desorbed to obtain a P equilibrium concentration of 0 mg l⁻¹) (2–75 mg kg⁻¹), both correlating closely with Al‐bound P and the P saturation degree of Al oxides (DPS_Alox). Maximum P sorption (Q_max) (43–515 mg kg⁻¹) also correlated most closely with Al_ox. The indoor and outdoor rainfall simulations produced sediments with different P sorption properties: in the indoor simulation (less kinetic energy, constant slope), the sediments had larger EPC₀ values, and usually larger −Q₀ values, than the sediments in the outdoor simulation (greater kinetic energy, variable slopes). Furthermore, the P exchange properties of the sediments differed from those of the bulk soil depending on the enrichment of soil P‐sorption components (Fe/Al oxides, clay). The outdoor simulation indicated that sites with gentle slopes produced sediments that were more enriched with Al_ox, Fe_ox, Mn_ox and organic C than those with steeper slopes. In this study, when the bulk soil had an initial EPC₀ greater than 1.3 mg l⁻¹, the outdoor rainfall simulation produced sediment with smaller EPC₀ and vice versa, indicating that, depending on the P status of the bulk soil, the sediment material was acting as source or sink for P during transport. However, on the basis of their EPC₀ values, most eroding sediments might be expected to desorb, rather than adsorb, P when entering surface water.     

Development of a mouse model to determine the systemic activity of potential flea-control compounds.

Probe studies were performed to determine if the cat flea (Ctenocephalides felis), the most common ectoparasite of companion animals, will feed on laboratory mice and, if so, to incorporate this into a small animal assay to detect systemically active compounds. Consequently, a protocol was developed which incorporated acepromazine maleate to temporarily sedate various strains of mice and allow fleas a window of time to feed undisturbed. For validation of the model, CD-1 mice were dosed per os with seven known insecticides at 30, 10 and 1mg/kg. Mice were sedated with 0.0125 ml acepromazine maleate intraperitoneally, and infested with fleas. After 2h, fleas were removed, one-third were examined immediately to confirm the occurrence of feeding, and 77% were found to have ingested a blood meal. The remaining fleas were incubated for 24h to determine mortality. Nitenpyram, the active ingredient in Capstar, was highly active (>94%) at 1mg/kg. Selamectin, the active ingredient in Revolution, was very active (86%) at 10mg/kg, but inactive at 1mg/kg. Fipronil, the active ingredient of Frontline Topspot, was very active (83%) at 30 mg/kg, moderately active (54%) at 10mg/kg and inactive at 1mg/kg. Cythioate, the active ingredient in Proban, and nodulisporic acid, a recently discovered oral insecticide, were moderately active (64 and 55%, respectively) at 10mg/kg, but both were inactive at 1mg/kg. Lufenuron and ivermectin exhibited no efficacy at any level tested. These findings suggest that this mouse model can effectively identify systemic flea-control leads and, subsequently, reduce the use of large animals in research.     

Neoskrjabinolepis merkushevae sp. n. (Cyclophyllidea: Hymenolepididae), a new cestode from shrews from the Palaearctic region

Neoskrjabinolepis (Neoskrjabinolepidoides) merkushevae sp. n. is described from the shrew Sorex minutus L. (type host) from Belarus. The new species is characterised by rostellar hooks 35-37 microm long, provided with small epiphyseal thickening of the handle; short (35-40 microm) cirrus consisting of a basal part with claw-shaped spines, a parabasal part with thin needle-shaped spines and an unarmed distal part; cirrus-sac slightly passing into median field; and 12-16 eggs per gravid uterus. In addition to the type host and the type locality, the host range of the new species includes Sorex araneus L. and there are geographical records from Bulgaria, Lithuania and North-East Altay (Russia).     

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 .     

Simple Technologies for Converting Rest Raw Materials of Atlantic Salmon (Salmo salar) into High-Quality,Valuable, and Tasty Feed Ingredients

Rest raw materials as viscera, heads, and frames from farmed Atlantic salmon (Salmo salar) were hydrolyzed with the use of endogenous enzymes and the commercial enzymes Protamex and a mixture of Papain and Bromelain. Composition of the prehydrolysis mixture clearly influenced the quality of final hydrolysate and process kinetics. An increased proportion of viscera increased the amount of endogenous enzymes, which influenced hydrolysis kinetics, the extent of hydrolysis, and increased bitterness. Commercial enzymes, in addition to endogenous enzymes, are not always more yield efficient or economically beneficial but can be used to improve the taste and to ease the separation of oil from the hydrolysate fraction. Hydrolysis of denatured proteins in rest raw materials was hardly detectable. Optimum temperature should therefore be selected to avoid protein denaturation and simultaneously maintain high enzymatic activity. Hydrolysates preferably contain a low concentration of oil, and this work shows that high-quality oil can be separated before hydrolysis by mild thermal treatment. The previous separation of oil did not influence hydrolysate yield and decreased the concentration of lipids in the final hydrolysate. The initial separation of oil also increased productivity of the hydrolysis reactor, due to the reduction of hydrolysis volume.     

Effects of organic and conventional fertilization on the occurrence of Brevicoryne brassicae L. and its natural enemies in white cabbage

Abstract

Recent years have witnessed a rapid growth in organic farming in Lithuania. We aimed to compare the occurrence of Brevicoryne brassicae L., its primary parasitoid Diaeretiella rapae (McIntosh) and predators from Coccinellidae and Cecidomyiidae families in non-fertilized, organically (with cow manure) and synthetically fertilized white cabbage crops. Occurrence of B. brassicae was higher on organically fertilized plants. Parasitoid D. rapae had the strongest effect in reducing the aphid population, as other predators occurred only in small densities. Quantity of parasitized aphids by D. rapae was higher on manure fertilized plants. In non-fertilized cabbage plants seasonal parasitization per cent of B. brassicae by D. rapae increased along with the expected lower aphid pressure compared with fertilized plants and reached on average 28.5%. D. rapae reduced the population of cabbage aphid by on average 15.5% in manure fertilized and ?12.9% in synthetically fertilized plants. The regression showed a positive relationship between the increase of the number of B. brassicae and activity of D. rapae. No coccinellids were found in aphid colonies on synthetically fertilized plants. Different fertilization did not have any significant effect on the incidence of Aphidoletes aphidimyza Rond. In conclusion, organic fertilization increased the population of cabbage aphid more than its natural enemies and the abundance of natural enemies was too small to provide effective reduction of this pest.     

Novel techniques for in vivo hemolysis studies in guinea pigs

The in vivo toxic-hemolytic studies using small experimental animals are complicated by difficulties in preventing hemolysis during repeated collection of blood specimens and in measuring hemoglobin concentration in small amounts of plasma sample. To solve these problems we tried to develope the new techniques for the in vivo hemolysis studies using guinea pigs. The hemolysis accident was minimized to 2.75 mg/dl by collecting the blood directly into heparinized microhematocrit tubes by small longitudinal incision in the auricular artery. The hemoglobin in a small amount of sample (10 microliters) was determined by the new analytical system using a microflow spectrophotometer with a modified cyanmethemoglobin method. The standard curve of the hemoglobin concentration in the system revealed a line of Y = 1.8X + 0.79 (r = 0.999), CV < 1% with a minimum detectable concentration of 1.25 mg/dl. By using the new techniques, it was found that the plasma hemoglobin concentration in normal animals were 7.27 +/- 0.44 mg/dl (mean +/- S.E.). The in vivo hemolytic activity of saponin was observed dose-dependently at doses of 30-50 mg/kg, i.v. in the guinea pigs. It is concluded that the present techniques are useful for in vivo hemolytic studies in small experimental animals such as guinea pigs.