Transformation of Petroleum Saturated Hydrocarbons during Soil Bioremediation Experiments

This paper presents transformations of saturated hydrocarbons of petroleum type pollutants during ex situ bioremediation of soil on the pilot heap (halde), during a period of 6 months, within the grounds of Petroleum Refinery Pan?evo (Serbia). Samples for analysis were taken in time intervals of 2 weeks (P₁–P₁₂ samples). Organic substance was extracted by Soxhlet’s method and quantified. Isoprenoid aliphatics, in particular pristane and phytane, and polycyclic aliphatics of sterane and triterpane types in saturated hydrocarbon fractions were analysed by GC-MS (SIM method). Significant amounts of n-alkanes have not been detected. The MS-chromatogram revealed only marginal amounts of pristane and phytane in sample P₁. Pristane and phytane occurred in sample P₈, and in even higher quantities in the final sample P_12. The proceeding bioremediation process was accompanied by the decrease of the relative amounts of pentacyclic terpanes of hopane type, compared to tri- and tetracyclic terpanes. In the initial sample P₁ the distribution of steranes and hopanes follows a pattern, which is characteristic for crude oils. However, their identification by SIM method was not possible in samples P₈ and P₁₂ because of the reduced concentration. The observed changes in the alkane fractions’ compositions may be considered as atypical, referring to the fact that during oil biodegradation under natural conditions, decomposition of isoprenoids occurs much easier and faster than decomposition of polycyclic alkanes of tri-, tetra- and pentacyclic terpane, sterane and diasterane types, after the decomposition of n-alkanes has been almost completed.     

Molecular detection of Theileria sp. in ticks and naturally infected sheep

Seven healthy sheep and 10 sheep diagnosed with piroplasmosis based on clinical signs were tested for the presence of babesiae and theileriae. Using the molecular techniques, two species of theileriae were detected and characterized. Theileria ovis was present mostly in healthy sheep and in Rhipicephalus ticks collected from infected sheep. Theileria sp. OT3 parasite was detected mostly in ill animals which represent additional evidence to the possible pathogenic nature of Theileria sp. OT3. The presence of babesiae in sheep or in ticks was not determined. The results of this study showed that ovine piroplasmosis due to Theileria is present in Southern Croatia. It was concluded that clinical diagnosis of ovine piroplasmosis should be confirmed by molecular analysis in order to identify the species of piroplasm, to select the appropriate treatment and to exclude the threat for public health.     

Sequence analysis of both genome segments of three Croatian infectious bursal disease field viruses

In order to determine the mutations responsible for virulence, three Croatian field infectious bursal disease viruses (IBDV), designated Cro-Ig/02, Cro-Po/00, and Cro-Pa/98 were characterized. Coding regions of both genomic segments were sequenced, and the nucleotide and deduced amino acid sequences were compared with previously reported full-length sequenced IBDV strains. Phylogenetic analysis, based on the nucleotide and deduced amino acid sequences of polyprotein and VP1, was performed. Eight characteristic amino acid residues, that were common to very virulent (vv) IBDV, were detected on polyprotein: 222A, 256I, 294I, 451L, 685N, 715S, 751D, and 1005A. All eight were found in Cro-Ig/02 and Cro-Po/00. C-Pa/98 had all the characteristics of an attenuated strain, except for glutamine on residue 253, which is common for vv, classical virulent, and variant strains. Between less virulent and vvIBDV, three substitutions were found on VP5: 49 G --> R, 79 --> F, and 137 R --> W. In VP1, there were nine characteristic amino acid residues common to vvwIBDV: 146D, 147N, 242E, 390M, 393D, 511S, 562P, 687P, and 695R. All nine residues were found in A-Ig/02, and eight were found in B-Po/00, which had isoleucine on residue 390. Based on our analyses, isolates Cro-Ig/02 and Cro-Po/00 were classified with vv IBDV strains. C-Pa/98 shared all characteristic amino acid residues with attenuated and classical virulence strains, so it was classified with those.     

High prevalence of intestinal zoonotic parasites in dogs from Belgrade, Serbia--short communication

To identify areas of risk for canine-related zoonoses in Serbia, the aim of this study was to provide baseline knowledge about intestinal parasites in 151 dogs (65 household pets, 75 stray and 11 military working dogs) from Belgrade. The following parasites, with their respective prevalences, were detected: Giardia duodenalis (14.6%), Ancylostomatidae (24.5%), Toxocara canis (30.5%), Trichuris vulpis (47.0%) and Taenia-type helminths (6.6%). Of all examined dogs, 75.5% (114/151) were found to harbour at least one parasite species. Of these, mixed infections with up to four species per dog occurred in 44.7% (51/114). Infections with all detected species were significantly higher (p < 0.05) in military working (100%) and stray dogs (93.3%) versus household pets (50.8%). Among all parasites, agents with zoonotic potential including Giardia, Ancylostomatidae and Toxocara were detected in 58.3% (88/151) of all examined dogs with a significant difference (p < 0.05) among the subgroups (100%, 62.7% and 46.2% for military working dogs, stray dogs and household pets, respectively). The high prevalence of zoonotic parasites registered in the dog population from a highly urban area in south-eastern Europe indicates a potential risk to human health. Thus, veterinarians should play an important role in helping to prevent or minimise zoonotic transmission.     

Plasma parathyroid hormone-related peptide and bone metabolism in periparturient dairy cows

The first weeks of lactation in dairy cows are characterised by elevated bone resorption. The connection between lactation and bone metabolism is still much discussed. In this work, changes in the concentration of plasma parathyroid hormone-related peptide (PTHrP) and markers of bone metabolism were studied in Holstein cows and heifers in the dry period and early lactation to determine the role of PTHrP in the relationship between the rate of bone remodelling and the onset of lactation in dairy cows. Blood samples were taken 14 days before calving ('D-14', n = 23) and then on day 10 ('D+10', n = 21) and day 30 after calving ('D+30', n = 23). Using enzyme immunoassay (EIA), the concentrations of PTHrP, parathyroid hormone (PTH), carboxyterminal cross-linked telopeptide of type I collagen (CTX) and oestradiol and the activity of bone specific alkaline phosphatase (BSALP) were determined. The results showed a statistically significant increase in plasma PTHrP (p < 0.005) and CTX (p < 0.0001) in cows on 'D+10' as compared to 'D-14' and CTX on 'D+30' as compared to 'D-14' (p < 0.0001). Significant negative correlations were found between the concentrations of PTHrP and oestradiol (r = -0.29, p < 0.05) and those of CTX and oestradiol (r = -0.54, p < 0.0001). In nonpregnant heifers (n = 6), the concentration of CTX and the activity of BSALP were significantly higher (p < 0.0001) than in dry cows. The observed increments of PTHrP and bone resorption after parturition reveal adaptations of bone metabolism to lactation in dairy cows.     

Comparison of mouse and rabbit model for the assessment of strong PGM-containing oil-based adjuvants

Peptidoglycan monomer (PGM) is an adjuvant active molecule with potential for use in human and veterinary vaccine. PGM's action is short-lived in mammals hence its effects might be limited. Novel PGM-containing oil-based formulations have been developed recently by incorporation of PGM into Montanide ISA720 and ISA206 adjuvants with the aim to prolong and improve immunostimulating activities of PGM. In the present work we studied the efficacy of such novel adjuvant formulations using two different antigens, ovalbumin and snake venom, respectively. Novel formulations were also tested in two experimental models, mice and rabbits. In rabbits the incorporation of PGM into oil-based adjuvants led to overall improvement of antigen-specific IgG response. However, in the mouse model, under experimental conditions used, it was not possible to distinguish differences in antigen-specific IgG response among several strong oil-based adjuvant formulations.     

N. femoralis, N. obturatorius and N. ischiadicus: deviation in creation in the dogs

Two hundred and twelve dog cadavers belonging to different breeds were examined, to investigate the formation of the femoral, obturator and the sciatic nerve. Besides the commonly described formation patterns of the mentioned nerves, some variations were also found. These variations were not gender-related, but on the other hand we discovered a certain correlation between the variations appearing in the formation of the femoral, obturator and the sciatic nerve. In 74.05% of cases, the femoral nerve was formed from ventral branches of the 4th, 5th and 6th lumbar nerve, and 16.98% of the dogs had the nerve formed from ventral branches of the 3rd, 4th and 5th lumbar nerve. Many dogs (i.e. 2.30%) had the femoral nerve formed from the ventral branches of the 5th, 6th and 7th, the 3rd, 4th, 5th and 6th or the 4th, 5th, 6th and 7th lumbar nerve, respectively. In 1.88% of dogs in particular, the femoral nerve was formed from ventral rami of the 4th and 5th lumbar nerve. In 66.98% of the examined dogs, the obturator nerve was formed from the ventral branches of the 4th, 5th and 6th lumbar nerve, followed by 16.59% of the dogs with the obturator nerve formed from the ventral rami of the 4th and 5th lumbar nerve and 9.43% of dogs in which the nerve was formed from the ventral branches of the 5th and 6th lumbar nerve. In 4.71% of dogs, the obturator nerve was formed from the ventral rami of the 4th, 5th, 6th and 7th lumbar nerve, while only 2.30% of the examined dogs had the same nerve formed from the ventral branches of the 5th, 6th and 7th lumbar nerve. The sciatic nerve was formed from ventral branches of the 6th and 7th lumbar nerve and the 1st sacral nerve in 86.79% of the dogs. In 5.18% of cases, the same nerve was formed from a junction of the ventral branches of the 7th lumbar and the 1st and 2nd sacral nerve, and, in the same percentage of cases, it was formed from a junction of ventral branches of the 6th and 7th lumbar nerve and the 1st and 2nd sacral nerve. In 2.83% of the dogs, it was formed from a junction of the ventral branches of the 6th and 7th lumbar and the 1st sacral nerve. The correlation of variations established in the formation of the femoral, obturator and the sciatic nerve was not statistically significant.     

Evidence of scientific research on organic plant breeding: A bibliometric study

Use of varieties bred under organic conditions is essential in order to minimize the yield gap between organic and conventional agriculture. The aim of this study was to analyse research publications related to the topic ‘organic plant breeding’ from the Web of Science database using bibliometric science mapping and visualization tools. The number of analysed documents in the bibliographic dataset was 204 from the 53 sources. The overall trend in the organic plant breeding literature showed that the number of publications increased during the observed time-span. We found that in total, 65 countries and 337 institutions are active in the field of organic plant breeding with a high degree of international collaboration. The top five countries according to the number of publications were the United States, Italy, Germany, France, and Canada, while the most active institutions were Wageningen University, Iowa State University, University of Alberta, University of Copenhagen, and University of Hohenheim. All keywords from the organic plant breeding research in the agronomy category were separated into seven clusters for different research topics. Although there is evident progress viewed through the increased trend in the number of publications, organic plant breeding needs further expansion and development. This is especially through the implementation of novel plant breeding techniques and methods aiming to improve traits that are highly specific to organic conditions.