Volume 8, Issue 6

Type 1 Diabetes mellitus (T1DM) begins with aberrant inflammatory process followed by auto-destruction in genetically susceptible individuals. Therefore, we hypothesized that gain-of-function allelic variants TNF-α-238A, -308A and PTPN22 1858T could be associated not only with T1DM development but also with the clinical outcome in patients of Bosnia and Herzegovina. A total of 402 subjects were enrolled in the association study. SNPs were determined by PCR-RFLP. Data was analyzed by GraphPad Prism and Sigma Stat 3.5 software. Genotypes frequencies at TNF-α-238 and -308 loci were not statistically different between patients and controls. In contrast, distribution of genotypes at the 1858 position of PTPN22 was significantly different, due to higher frequency of gain-of-function gene variants in patients than controls. Moreover, long term glucose regulation (based on HbA1c level) was significantly worse in patients with the risk TNF-α-308A allele than in patients with non-risk (G) allele. However, patients with the risk allele of both genes (TNF-α-308A and PTPN22 1858T) had the worst glycemic control, suggesting that those two work synergistically. In conclusion, in a cohort from Bosnia and Herzegovina TNF-α-308A allele is significantly associated with the worse long-term glucose control, but PTPN22 1858T allele is significantly associated with diabetes development.

Newcastle disease Virus (NDV), a member of the Paramyxoviridae family, and Influenza virus, from the Orthomyxoviridae family, are two main avian pathogens that cause serious economic problems in poultry farming. NDV strains are classified into three major pathotypes: velogenic, mesogenic, and lentogenic. Avian influenza viruses (AIV) are also divided into: low pathogenic (LPAI) and highly pathogenic (HPAI) strains. Both viruses are enveloped, single stranded, negative-sense RNA viruses which give similar symptoms ranging from sub-clinical infections to severe disease, including loss in egg production, acute respiratory syndrome, and high mortality, depending on their level of pathogenicity. This similarity hinders diagnosis when based solely on clinical and post mortem examination. Most of the currently available molecular detection methods are also pathogenspecific, so that more than one RT-PCR is then required to confirm or exclude the presence of both pathogens. To overcome this disadvantage, we have applied a One Step Duplex RT-PCR method to distinguish between those two pathogens. The main objective of the project was to develop a universal, fast, and inexpensive method which could be used in any veterinary laboratory.

The aim of the study was to determine if there is a relationship between serum selenium concentration in dogs and their health, and to assess the relationship between selenium concentration and morphological parameters of blood. Mean serum selenium concentration in dogs ranged from 0.169 to 0.273 mg/ml. Dogs diagnosed with malignant neoplasm had a significantly lower mean concentration of serum selenium compared to healthy dogs and those from the groups studied. The present study showed no statistically significant differences in Se serum content according to sex, age, and food type. Dogs diagnosed with malignant neoplasm had a significantly lower mean serum selenium concentration compared to healthy dogs and those from the other groups analysed, namely with hip dysplasia, allergy and fractures and with non-malignant tumour. Healthy dogs were characterized by the highest mean serum selenium concentration, significantly higher compared to dogs with non-malignant tumour, malignant neoplasm, allergy and fractures. Low levels of selenium contribute to the incidence of neoplasms and allergies and increase the risk of bone fractures in dogs. Additional laboratory tests should be conducted when certain diseases are diagnosed to determine Se concentration in dogs, thus making it possible to take preventive measures or therapeutic action.

Taxonomy of thermophilic, endospore-forming bacteria has evoked a great interest over the past few years. Although a number of taxonomic markers were previously evaluated, their sequences in Geobacillus were too conservative, and identification of more variable markers is needed. Repetitive DNA is one of the promising variable targets in the development of the taxon-specific genotyping and identification schemes in bacteria. The aim of our study was to evaluate the possibility of using repetitive DNA in the taxonomy of Geobacillus. In this paper, we report the analysis of perfect tandem repeats of geobacilli. We focused on the long repeats (with a motif length of ≥20 nucleotides). This choice was based on the assumption that these motifs can be used for the construction of oligonucleotides — primers and probes. Thirty-three Geobacillus genus-specific motifs were identified in our work, fifteen of them were species-specific and fifteen — species cluster -specific. Three of them were genus-, but not species- or species cluster-specific. Some of the motifs were used for the construction of the primer pairs. The primers were validated by PCR. Out of 12 designed primer pairs, 11 were genus-specific and 4 — species-specific. Species-specific primers were successfully constructed for the phylogenetically defined species Geobacillus thermodenitrificans and Geobacillus toebii.

Insects are good models for studying the innate immune response. We report that Galleria mellonella larvae infected with entomopathogenic bacteria Bacillus thuringiensis kurstaki show changes in the level of Hsp90. Our experimental approach was to pre-treat larvae with the Hsp90-binding compound, 17-DMAG, before infection with B. thuringiensis. We show that pre-treated animals display a higher level of immune response. This was mainly manifested by enhanced action of their hemolymph directed toward living bacteria as well as lysozyme activity digesting bacterial peptidoglycan. The observed phenomenon was due to the higher activity of antimicrobial peptides which, in contrast to healthy animals, was detected in the hemolymph of the immunestimulated larvae. Finally, the physiological significance of our observation was highlighted by the fact that G. mellonella pre-treated with 17-DMAG showed a prolonged survival rate after infection with B. thuringiensis than the control animals. Our report points to a role for Hsp90 in the immune response of G. mellonella after infection with B. thuringiensis at the optimal growth temperature.

Aphids are a good model to study insect reaction to habitat change. Temperature is one of the main factors that influences insects. This paper examines the influence of temperature on developmental stages, fecundity, survival rate and demographic parameters of Cinara tujafilina (Hemiptera: Aphidoidea, Lachnidae), connected with decorative plants of the Cupressaceae family. C. tujafilina was reared in a laboratory on T. orientalis at five constant temperatures of 10, 15, 20, 25 and 28°C, 70% humidity and 14L:10D. The pre-reproduction stage varied from 7 at 25°C to 19 days at 10°C. Developmental threshold was assigned at 3.5°C. The longest reproduction stage for the aphids developing was recorded at 25°C, namely 33 days, while the shortest, at the temperature of 10°C, lasted 8 days. At 25°C this species is characterised by the shortest pre-reproduction stage, the highest fecundity, the highest survival rate and the highest demographic parameters, particularly rm (0.17). The results suggest that the optimal temperature for the species is 25°C, and indicate that climatic change will favourably influence its development and increase its role as a pest of decorative plants.

Microalgae are an alternative and sustainable source of lipids that can be used as a feedstock for biodiesel production. Nitrate is a good nitrogen source for many microalgae and affects biomass and lipid yields of microalgae. In this study, the effect of nitrate on cell growth and lipid production and composition in Monoraphidium contortum, Tetraselmis suecica, and Chlorella minutissima was investigated. Nitrate affected the production of biomass and the production and composition of lipids of the three microalgae tested. Increasing the nitrate concentration in the culture medium resulted in increased biomass production and higher biomass productivity. Furthermore, increasing the nitrate concentration resulted in a reduction in lipid content and productivity in M. contortum; however, the opposite effect was observed in T. suecica and C. minutissima cultures. C. minutissima and M. contortum lipids contain high levels of oleic acid, with values ranging from 26 to 45.7% and 36.4 to 40.1%, respectively. The data suggest that because of its high lipid productivity (13.79 mg L−1 d−1) and high oleic acid productivity (3.78 mg L−1 d−1), Chlorella minutissima is a potential candidate for the production of high quality biodiesel.

In vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

In this study the glandular hair morphology, chemical composition and antimicrobial activity of the essential oils from three Ocimum species have been investigated (Ocimum basilicum L. var. Genovese, O. gratissimum and O. tenuiflorum). The indumentum shows little variation among the investigated species with both glandular and non-glandular hairs presents. Glandular hairs on the three species are peltate and capitate (with various cell numbers in the stalk and gland). The samples of essential oils obtained from the plant aerial organs by hydrodistillation have been analyzed by GC-MS. Linalool (65.38%, 74.22%, 38.60%), eugenol (5.26%, 3.47%, 10.20%) and tau-cadinol (8.18%, 3.47%, 10.20%) appear as the main components in Ocimum basilicum L. var. Genovese, O. gratissimum and O. tenuiflorum. The oils also contain lower levels of α-bergamotene, 1,8-cineole, germacrene D, β-ocimene, α-caryophyllene, camphor, and α-guaiene. All essential oils showed antibacterial activity against Staphylococcus aureus and Escherichia coli depending on their concentration. Ocimum basilicum L. var. Genovese oil produced the strongest antibacterial effect on S. aureus and E. coli.