Influence of increased lead concentration on changes in microbiota composition and life history traits in Drosophila melanogaster and Drosophila subobscura

Mikrobiota višećelijskih organizama je predstavljena raznovrsnom zajednicom mikroorganizama koji imaju uticaj na brojne osobine domaćina i u interakciji su sa promjenama sredine. Poremećaji u ravnoteži između mikrobiote i domaćina mogu dovesti do smanjenja adaptivne vrijednosti, što implicira da je održavanje homeostaze mikrobiote važan aspekt biologije domaćina. Cilj ovog istraživanja je bio da se utvrdi diverzitet mikrobiote vrsta Drosophila melanogaster i Drosophila subobscura sa različitih staništa iz prirode i u laboratorijskim uslovima na standardnom i olovnom supstratu, te da se ispita korelacija promjena u sastavu mikrobiote sa osobinama životne istorije. Sastav mikrobiote je određen sekvenciranjem sljedeće generacije V3-V4 regiona bakterijskog gena za 16S rRNK. Dobijeni rezultati ukazuju na to da standardni supstrat gajenja kod obje vrste dovodi do smanjenja diverziteta mikrobiote u odnosu na prirodnu populaciju. Izloženost povišenim koncentracijama olova je dovela do smanjenja diverziteta mikrobiote, pri čemu je D. subobscura pokazala značajnije promjene u odnosu na D. melanogaster. U mikrobioti D. subobscura je identifikovan bakterijski rod Komagataeibacter, čija velika zastupljenost na supstratu sa olovom ukazuje na to da bi ovaj rod mogao biti značajan u prevazilaženju stresa iz životne sredine, kao i da postoji kontrola sastava mikrobiote od strane domaćina. Značajne razlike u preživljavanju od jaja do adulta su uočene kod D. melanogaster u pogledu sastava supstrata i porijekla populacije, dok je dužina razvića značajno varirala kod D. subobscura u pogledu porijekla populacija. Razlike među polovima u dužini razvića su uočene kod D. melanogaster na olovnom supstratu, što implicira da uticaj pojedinih faktora na osobine životne istorije može biti specifičan za vrstu.Microbiota of multicellular organisms is represented by a diverse community of microorganisms that have an impact on numerous host traits and interact with environmental changes. Disturbances in the balance between the microbiota and the host can lead to a decrease in adaptive value, which implies that maintaining microbiota homeostasis is an important aspect of host biology. The aim of this study was to determine the diversity of microbiota of Drosophila melanogaster and Drosophila subobscura from different habitats, nature and laboratory conditions, on the standard and the lead substrate, and to examine the correlation between the changes in microbiota composition and life history traits. The composition of microbiota was determined by next-generation sequencing of the V3-V4 regions of the 16S rRNA gene. The obtained results indicate that rearing on the standard substrate in both species leads to a decrease in the diversity of the microbiota compared to the natural population. Exposure to increased lead concentrations led to a decrease in microbiota diversity, with D. subobscura showing more significant changes compared to D. melanogaster. The bacterial genus Komagataeibacter was identified in the D. subobscura's microbiota, whose high prevalence on the lead substrate indicates that this genus could be important in overcoming environmental stress, and that the microbiota composition is controlled by the host. Significant differences in egg-to-adult viability were observed in D. melanogaster in terms of substrate composition and population origin, while developmental time varied significantly in D. subobscura in terms of population origin. Sex differences in developmental time were observed in D. melanogaster on the lead substrate, implying that the influence of individual factors on life history traits may be species-specific

COMPARISON OF TWO DIFFERENT MULTIPLEX SYSTEMS IN CALCULATING KINSHIP AMONG CLOSE RELATIVES

This study compares the results obtained using two multiplex systems, PowerPlex® 16 System and PowerPlex® Fusion System, to evaluate the probability of a specific kinship relationship between the offspring of three pairs of identical twins, such as full kinship (siblings), first-degree relatives (first cousins) and half-siblings. Genomic DNA was isolated and amplified from buccal swab and selected short tandem repeat (STR) markers were detected. Electropherograms were generated and analyzed for all persons, using two multiplex systems. Paternity testing for every nine offspring of six examined couples was performed and in all cases the probability that the alleged father is the true father, was over 99.9999%. Kinship analyses were performed setting up two different hypotheses and calculating the likelihood ratio (LR) and kinship probability. Determining the degree of kinship between persons who were full siblings, likelihood ratio showed the highest values contrary to other two types of kinship. Kinship analyses between first cousins showed a higher probability that the examined persons are half-siblings, rather than they are first cousins. In most cases, the introduction of additional seven loci included in PowerPlex® Fusion System increased the values of average likelihood ratios. It is recommendable to use over 20 STR loci in complex kinship analyses

Life History Traits in Two Drosophila Species Differently Affected by Microbiota Diversity under Lead Exposure

Life history traits determine the persistence and reproduction of each species. Factors that can affect life history traits are numerous and can be of different origin. We investigated the influence of population origin and heavy metal exposure on microbiota diversity and two life history traits, egg-to-adult viability and developmental time, in Drosophila melanogaster and Drosophila subobscura, grown in the laboratory on a lead (II) acetate-saturated substrate. We used 24 samples, 8 larval and 16 adult samples (two species × two substrates × two populations × two sexes). The composition of microbiota was determined by sequencing (NGS) of the V3–V4 variable regions of the 16S rRNA gene. The population origin showed a significant influence on life history traits, though each trait in the two species was affected differentially. Reduced viability in D. melanogaster could be a cost of fast development, decrease in Lactobacillus abundance and the presence of Wolbachia. The heavy metal exposure in D. subobscura caused shifts in developmental time but maintained the egg-to-adult viability at a similar level. Microbiota diversity indicated that the Komagataeibacter could be a valuable member of D. subobscura microbiota in overcoming the environmental stress. Research on the impact of microbiota on the adaptive response to heavy metals and consequently the potential tradeoffs among different life history traits is of great importance in evolutionary research

Influence of increased lead concentration on changes in microbiota composition and life history traits in Drosophila melanogaster and Drosophila subobscura

Mikrobiota višećelijskih organizama je predstavljena raznovrsnom zajednicom mikroorganizama koji imaju uticaj na brojne osobine domaćina i u interakciji su sa promjenama sredine. Poremećaji u ravnoteži između mikrobiote i domaćina mogu dovesti do smanjenja adaptivne vrijednosti, što implicira da je održavanje homeostaze mikrobiote važan aspekt biologije domaćina. Cilj ovog istraživanja je bio da se utvrdi diverzitet mikrobiote vrsta Drosophila melanogaster i Drosophila subobscura sa različitih staništa iz prirode i u laboratorijskim uslovima na standardnom i olovnom supstratu, te da se ispita korelacija promjena u sastavu mikrobiote sa osobinama životne istorije. Sastav mikrobiote je određen sekvenciranjem sljedeće generacije V3-V4 regiona bakterijskog gena za 16S rRNK. Dobijeni rezultati ukazuju na to da standardni supstrat gajenja kod obje vrste dovodi do smanjenja diverziteta mikrobiote u odnosu na prirodnu populaciju. Izloženost povišenim koncentracijama olova je dovela do smanjenja diverziteta mikrobiote, pri čemu je D. subobscura pokazala značajnije promjene u odnosu na D. melanogaster. U mikrobioti D. subobscura je identifikovan bakterijski rod Komagataeibacter, čija velika zastupljenost na supstratu sa olovom ukazuje na to da bi ovaj rod mogao biti značajan u prevazilaženju stresa iz životne sredine, kao i da postoji kontrola sastava mikrobiote od strane domaćina. Značajne razlike u preživljavanju od jaja do adulta su uočene kod D. melanogaster u pogledu sastava supstrata i porijekla populacije, dok je dužina razvića značajno varirala kod D. subobscura u pogledu porijekla populacija. Razlike među polovima u dužini razvića su uočene kod D. melanogaster na olovnom supstratu, što implicira da uticaj pojedinih faktora na osobine životne istorije može biti specifičan za vrstu.Microbiota of multicellular organisms is represented by a diverse community of microorganisms that have an impact on numerous host traits and interact with environmental changes. Disturbances in the balance between the microbiota and the host can lead to a decrease in adaptive value, which implies that maintaining microbiota homeostasis is an important aspect of host biology. The aim of this study was to determine the diversity of microbiota of Drosophila melanogaster and Drosophila subobscura from different habitats, nature and laboratory conditions, on the standard and the lead substrate, and to examine the correlation between the changes in microbiota composition and life history traits. The composition of microbiota was determined by next-generation sequencing of the V3-V4 regions of the 16S rRNA gene. The obtained results indicate that rearing on the standard substrate in both species leads to a decrease in the diversity of the microbiota compared to the natural population. Exposure to increased lead concentrations led to a decrease in microbiota diversity, with D. subobscura showing more significant changes compared to D. melanogaster. The bacterial genus Komagataeibacter was identified in the D. subobscura's microbiota, whose high prevalence on the lead substrate indicates that this genus could be important in overcoming environmental stress, and that the microbiota composition is controlled by the host. Significant differences in egg-to-adult viability were observed in D. melanogaster in terms of substrate composition and population origin, while developmental time varied significantly in D. subobscura in terms of population origin. Sex differences in developmental time were observed in D. melanogaster on the lead substrate, implying that the influence of individual factors on life history traits may be species-specific

Organization of Laboratory for Monitoring Security in the Food Industry in Order to Detect the Presence of Allergens

Allergens are substances that cause allergic reactions. Allergic reactions differ from person to person in a sensitive and specific response to the presence of the same allergen. Groceries that often cause allergies are cow’s milk, eggs, fish, crustaceans and shellfish, wheat, soy, peanuts, walnuts, almonds, hazelnuts and strawberries.Organisation is the main factor for the success and the quality of a research in food industry laboratories, in order to detect the presence of allergens. All kinds of equipments are needed, as well as professional staff to perform the tests. Allergen testing in the food industry is often performed using biochemical and separation methods. For analysis of deoxyribonucleic acid (DNA), the most suitable method is polymerase chain reaction (PCR) and electrophoresis. In our laboratory, we use immunological methods for qualitative and quantitative testing of allergens and we have two accredited methods: Enzyme-Linked Immunosorbent Assays (ELISA) and High Performance Liquid Chromatography (HPLC). It is also necessary that stuff have adequate competence in handling the specific equipment, performing tests, evaluating the results and signing test reports and calibration certificates, have adequate competences. Laboratory have to prove that have been fulfiled all the requirements for validation. Validation includes: specification of requirements, characterization of method, verification that requirements can be fulfilled using the method.The results of each test are presented in form of a report, which has to be correct, clear, unambiguous, objective and must include all the informations required by the client

Estimation of antioxidative potential of thyme (Thymus alpestris L.)

Family Lamiaceae (labia) includes a large number of cosmopolitan speciesincluding Thymus alpestris (Thyme) characterized by the presence of essential oilsand phenolic compounds. It represents a perennial semi-shrubby plant. Theantioxidant potential of thyme is due to the presence of polyphenolic acids andflavonoids. The topic of this paper is the extraction of thyme (Thymus alpestris)with 40% ethanol and 50% ethanol, using Soxhlet method (circulatoryextraction), after which, the content of phenolics and flavonoids was determinedin order to confirm the antioxidant potential of this plant species. The resultsshow that 40% ethanol extract has the highest content of phenolics andflavonoids and the highest antioxidant potential, resultingly

Association of Dietary Patterns with MRI Markers of Hepatic Inflammation and Fibrosis in the MAST4HEALTH Study

Whereas the etiology of non-alcoholic fatty liver disease (NAFLD) is complex, the role of nutrition as a causing and preventive factor is not fully explored. The aim of this study is to associate dietary patterns with magnetic resonance imaging (MRI) parameters in a European population (Greece, Italy, and Serbia) affected by NAFLD. For the first time, iron-corrected T1 (cT1), proton density fat fraction (PDFF), and the liver inflammation fibrosis score (LIF) were examined in relation to diet. A total of 97 obese patients with NAFLD from the MAST4HEALTH study were included in the analysis. A validated semi-quantitative food frequency questionnaire (FFQ) was used to assess the quality of diet and food combinations. Other variables investigated include anthropometric measurements, total type 2 diabetes risk, physical activity level (PAL), and smoking status. Principal component analysis (PCA) was performed to identify dietary patterns. Six dietary patterns were identified, namely “High-Sugar”, “Prudent”, “Western”, “High-Fat and Salt”, “Plant-Based”, and “Low-Fat Dairy and Poultry”. The “Western” pattern was positively associated with cT1 in the unadjusted model (beta: 0.020, p-value: 0.025) and even after adjusting for age, sex, body mass index (BMI), PAL, smoking, the center of the study, and the other five dietary patterns (beta: 0.024, p-value: 0.020). On the contrary, compared with low-intake patients, those with medium intake of the “Low-Fat Dairy and Poultry” pattern were associated with lower values of cT1, PDFF, and LIF. However, patients with a “Low-Fat Dairy and Poultry” dietary pattern were negatively associated with MRI parameters (cT1: beta: −0.052, p-value: 0.046, PDFF: beta: −0.448, p-value: 0.030, LIF: beta: −0.408, p-value: 0.025). Our findings indicate several associations between MRI parameters and dietary patterns in NAFLD patients, highlighting the importance of diet in NAFLD