Study of the genetic variability of porcine circovirus type 2 detected in Serbia and Slovenia

Recent variants of porcine circovirus type 2 (PCV2) were obtained from tissues of domestic pigs with porcine circovirus associated disease and from randomly selected wild boar samples from Serbia and Slovenia. A 450-base-pair nucleotide sequence was obtained by PCR from the ORF2. The derived nucleotide and amino acid sequences were aligned and compared to the corresponding region of closely related PCV2 sequences determined in previous years and retrieved from the GenBank. The 30 Serbian and 17 Slovenian PCV2 sequences clustered into three previously determined genotypes (PCV2a: 7), (PCV2b: 38) and (PCV2d: 2). Three major variable regions, concerning 29 amino acid position substitutions within the ORF2, were observed, which further supports the segregation of the detected strains into three separate genotypes. This study indicates that PCV2b is the predominant genotype in Serbia and Slovenia and the detected PCV2 strains are closely related to those previously described in Europe and in other parts of the world.     

Special issue “Deepen knowledge in plant pathology for innovative agroecology”

This study aimed to determine the genetic variability of isolates of rotting of pineapple fruitlet core in Brazil on the states of Paraiba, Pernambuco and Rio Grande do Norte, based on phylogenetic analysis of the RPB2 gene, morphocultural markers and aggressiveness of the isolates. The phylogenetic reconstruction of maximum parsimony and bayesian inference of the isolates were performed. Twenty-seven phylogenetic lineages were characterized with morphocultural markers on potato dextrose agar and synthetic nutrient-poor agar. The aggressiveness of these lineages were characterized in leaves and fruits of pineapple ‘Pérola’ cultivar. The Phylogenetic reconstruction showed close relationship between isolates of this study and phylogenetic lineages of F. guttiforme, F. ananatum and F. oxysporum by analysis of RPB2 gene. Phylogenetic lineages of this study shared significant morphocultural markers similar to those described for this species. Overall, the lineages related with Fusarium oxysporum Species Complex were more aggressive to the fruits of the Fusarium lineages related with Fusarium fujikuroi Species Complex. It is possible that F. ananatum and F. guttiforme or other lineages closely related to these species or F. oxysporum are present in the fields causing pineapple rotting fruitlet core in fruits of pineapple ‘Pérola’ in Brazil. The used markers determined high genetic variability in isolates analyzed in leaves and pineapple fruit ‘Pérola’ in the states of Paraíba, Pernambuco and Rio Grande do Norte of Brazil and the pathogenic lineages analyzed were better adapted to the fruits than to the detached leaves in pineapple ‘Pérola’ analyzed.     

Chemical and nutritional characteristics of Cannabis sativa L. co-products

Cannabis sativa L. is an annual herbaceous plant. It was used for centuries to obtain different products. In the last century, hemp cultivation was forbidden due to the psychoactive effects of tetrahydrocannabinol acid (THCA). In the last years, new strains, characterized by high cannabidiolic acid (CBDA) and low THCA level, were developed renewing the interest in hemp cultivation to obtain food or to extract essential oils from flowers. All these processes produce many residues with different chemical–physical characteristics. In order to evaluate their potential use also in animal nutrition, some hemp co-products were evaluated. Two different co-products of seed processes (flour and oil) and two co-products obtained trimming the flowers, differing in granulometry were used. The samples were analysed for chemical composition and evaluated in vitro using the gas production technique with buffaloes' ruminal inoculum. All hemp co-products showed interesting nutritional characteristics, such as crude protein content always higher than 20% on a dry matter basis, and high neutral detergent fibre concentration partially lignified. The in vitro gas production parameters at 120 h of incubation showed quite low fermentability testified by the low organic matter degradability and cumulative gas volume (OMD from 28.09 to 45.64% and OMCV from 110 to 164 ml/g, respectively). Also, the methane produced after 24 h of incubation was particularly low (from 1.78 to 11.73 ml/g dOM). These results could be due to the high lipid and ash amounts or to the CBDA content that probably affected the CH₄ formation processes. According to preliminary results obtained by this study, it is possible to hypothesize that these co-products could be useful to mitigate the methane production into the rumen. Further studies are necessary in order to evaluate the correct inclusion into the diet for ruminants.     

Effects of bentonite on wool growth and nitrogen metabolism in fauna-free and faunated sheep.

Two experiments were carried out with sheep that originated from a fauna-free flock and were fed a soybean meal-corn silage diet with or without a bentonite supplement. One-half of the sheep fed each diet in each experiment were faunated with a mixed population of ruminal protozoa, whereas the other half of the sheep remained fauna-free until the end of both experiments. Wool growth and daily gain were measured in Exp. 1. (eight rams per treatment), which lasted 110 d, and the metabolic effects in the rumen and intestinal tract of protozoa and dietary bentonite supplement were tested with cannulated wethers (four wethers per treatment) in Exp. 2. The results of Exp. 1 showed decreased wool growth (P less than .05) due to the presence of protozoa in the rumen. Dietary supplementation with bentonite partly offset the decreased wool growth in sheep with protozoa, but there were no effects of dietary bentonite and no protozoa x bentonite interaction (P greater than .05). Daily gain was decreased by the dietary bentonite (P less than .05) supplement but was not affected (P greater than .05) by the ruminal presence of protozoa. In Exp. 2, protozoa increased (P less than .01) the ruminal concentrations of ammonia and decreased (P less than .05) the acetic:propionic acid molar ratio. Fractionation of N in the duodenal digesta flowing from the stomach to the small intestine showed that protozoa decreased (P less than .05) the flow of nonammonia N and bacterial N, and there was a protozoa x bentonite interaction for these effects (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Contamination history of suspended river sediments accumulated in oxbow lakes over the last 25 years

Background, aims, and scope  Embankment of meandering river systems in many industrial areas results in the formation of artificial oxbow lakes that may act as perennial or intermittent traps for river sediments. Their deposits can be dated using a combination of historical and stratigraphic data, providing a good means to study historical records of contamination transported by rivers. Contamination history over the last few decades is of special significance for Central and Eastern Europe as it can reflect high pollutant levels in the second half of the twentieth century and the subsequent improvement after the fall of the Iron Curtain. The purpose of this study was to investigate recent sediments of an oxbow lake of the Morava River, Czech Republic, their stratigraphic records, sediment architecture, and history of contamination. Materials and methods  Seven ground-penetrating radar (GPR) profiles and three sediment cores up to 4 m deep were studied. The stratigraphy of the cores was inferred from visible-light spectrophotometry, X-ray radiography, grain size analysis, and semiquantitative modal analysis of sandy fractions. The sediments were dated using the ¹³⁷Cs mass activity and combinations of stratigraphic and historical data. The cores were sampled for concentrations of heavy metals and persistent organic pollutants. Wet sampled, lyophilized, and sieved sediments were extracted and analyzed for heavy metals by inductively coupled plasma mass spectrometry (ICP-MS) of aqua regia leachate and for persistent organic pollutants by gas chromatography (GC-ECD and GC-MS). Results  Three distinct sedimentary sequences (S1, S2, and S3) were identified. The basal sequence S1 represents river channel sediments deposited before the formation of the oxbow lake, most likely before the 1930s. The boundary between the S1 and S2 sequence correlates with the level of sediment dredging from 1981 evidenced from historical data. The overlying sequences S2 and S3 represent a postdredging sediment wedge, which progrades into the lake. ¹³⁷Cs dating revealed a distinct Chernobyl 1986 peak at ∼150 cm depth inferring sedimentation rates up to 7.7 cm/year. Sediment contamination abruptly increased from the pre-1930s deposits to the post-1981 deposits. The concentration levels increased two to five times for Pb, As, Zn, and Cu, about 10 to 15 times for Cr, Sb, and Hg, up to 34 times for Cd, and 25 to 67 times for DDTs, PCBs, and PAHs. The concentrations of most contaminants remained approximately constant until the late 1980s when they started to decrease slowly. The decreasing trends were most prominent for heavy metals and anthracene, less prominent for DDTs, and almost absent for PCBs and most PAHs. Discussion  Different temporal and spatial patterns for various contaminants make it possible to distinguish between anthropogenic point sources from local industry (anthracene, Cr, Cd), possible diffuse sources (most PAHs), and geological background (V, Co, Ni, and Mo). The observed recent trends in heavy metal and persistent organic pollutant contamination are generally consistent with data from other Central European rivers. The roughly balanced contamination levels in sediments from the lake and the adjacent river channel suggest that the oxbow lake deposits reflect immediate levels of the contamination bound to suspended particulate matter passing through the river. Conclusions  The investigated oxbow lake accumulated suspended sediment from Morava River, developing a thick sedimentary body. The sediments offer a good time framework to study historical contamination of the river on a decade time scale. Continuous contamination trends can be traced back to the early 1980s. The results show that stratigraphic analysis of cores has a good potential for identification of uninterrupted historical trends and unconformities, e.g., due to dredging. Recommendations and perspectives  Oxbow lakes may provide an alternative to floodplains and reservoir deposits when studying river contamination history. High sedimentation rates of up to 7.7 cm/year offer a very good stratigraphic resolution, making it possible to study contamination patterns on annual or even seasonal time scales. On the other hand, oxbow lakes may represent long-term contamination stores, which are unlikely to be redistributed by river erosion and, hence, may possess significant environmental risks for the farther future.     

Use of soybean peroxidase in chemiluminescent enzyme-linked immunosorbent assay

A procedure for the production of conjugates of soybean peroxidase (SbP) oxidized by sodium periodate and anti-mouse IgG antibody (Ab) was optimized. A sandwich chemiluminescent enzyme-linked immunosorbent assay (ELISA) for determination of mouse IgG using SbP and specific Ab was developed, and SbP-catalyzed oxidation of luminol was carried out in the absence of any enhancer. Comparison of conjugates produced by labeling Ab by soybean and horseradish peroxidases in the chemiluminescent ELISA showed that in the case of SbP a rate of emission decay formed through luminol oxidation was significantly lower. Application of the soya enzyme allowed the development of the immunoassay with improved sensitivity and a wider linear range.     

Genetic typing of bovine viral diarrhoea virus: most Slovenian isolates are of genotypes 1d and 1f

A selection of 43 bovine viral diarrhoea viruses isolated from mainly persistently infected cattle on 23 Slovenian farms between 1997 and 2001 were characterised genetically. Viral RNA was extracted from infected cell cultures, reverse transcribed and amplified by PCR with primers targeting the 5'-UTR and the N(pro) gene, followed by direct sequencing of purified PCR products obtained for both genomic regions. The N(pro) sequences provided the best genetic resolution, and gave also higher statistical support for phylogenetic classification of the viruses. Thirty-eight of the Slovenian isolates were of genetic subtypes 1d and 1f, four were 1b, and one subtype 1g. No BVDV type 2 viruses were found. This genetic prevalence matched those previously reported for neighbouring countries, as opposed to findings reported for more distant European countries, e.g. France, Spain and the UK. From eight cattle herds several virus isolates were analysed; with one exception all isolates from each herd were of the same genetic group. Extended sequencing of the N(pro) and part of the C gene of virus isolates with identical 5'-UTR sequences allowed differentiation between isolates obtained at different times from one herd.