Elemental concentration in mealworm beetle (Tenebrio molitor L.) during metamorphosis.

Mealworm beetles have been used in numerous experiments as bioindicators. The aim of our experiment was to study the elemental composition in three larvae, pupae and first and second generation adult stages during their life cycle. We selected 180 larvae from a genetically similar population and put them in three groups, in two boxes (60 larvae in each box). Larvae were fed with mashed potato made of the same quality and quantity of potato powder. Then, we selected 10 individuals from each stage to the elemental analysis, using the ICP-OES method. The following elements were analysed in the studied stages: Ca, Cu, Fe, K, Mg, Mn, Na, P, S, Sr and Zn. The results of principal component analysis demonstrated that based on elemental composition, different stages were separated with each other, but in the cases of the three larvae stages, high overlap was found. The results of the GLM ANOVA showed significant differences between the different stages of metamorphosis-based elemental composition. Our results show that the calcium and magnesium were found in a relatively high concentration, while the iron and zinc may be essential elements during the metamorphosis. Our results also show that in insect, the concentration of sodium was higher than in the pupa which may cause by hemolymph. We also demonstrated that the metamorphosis has an effect on the concentration of elements. Our study shows that in the different stages of insects, there are significant changes in the elemental composition of different stages of insects during their metamorphosis.

Effect of high relative humidity on dried Plantago lanceolata L. leaves during long-term storage: effects on chemical composition, colour and microbiological quality.

INTRODUCTION: Modern phytotherapy and quality assurance requires stability data on bioactive metabolites to identify and minimise decomposing factors during processing and storage. A compound's stability in a complex matrix can be different from the stability of the purified compound. OBJECTIVE: To test the stability of iridoids and acteoside and quantify changes in colour and microbiological quality in a common herbal tea, dried P. lanceolata leaves during exposure to high-humidity air. To test the contribution of fungi to metabolite decomposition. METHODOLOGY: Dried P. lanceolata leaves were exposed to atmospheres of different relative humidity (75, 45 and 0%) for 24 weeks. Changes in aucubin and catalpol concentration were determined by CE-MEKC, and those in acteoside on TLC. Colour and chlorophyll-like pigments were measured by different spectrophotometric methods. The number of fungi was monitored; 10 strains were isolated from the plant drug, and their ability to decompose the analytes of interest was tested. RESULTS: During incubation at 75% relative humidity (RH), aucubin, catalpol and acteoside concentrations decreased by 95.7, 97.0 and 70.5%, respectively. Strong shifts were detected in CIELAB parameters a* and b* (browning) as a result of conversion of chlorophyll to pheophytin. Intensive microbial proliferation was also observed. Changes at 45 or 0% RH were typically insignificant. Seven of the 10 isolated fungal strains could decompose both iridoids, and five could decompose acteoside in vitro. CONCLUSION: It was shown that exposure to water results in loss of bioactive molecules of P. lanceolata dried leaves, and that colonising fungi are the key contributors to this loss.

Population dynamics and genetic changes of Picea abies in the South Carpathians revealed by pollen and ancient DNA analyses.

BACKGROUND: Studies on allele length polymorphism designate several glacial refugia for Norway spruce (Picea abies) in the South Carpathian Mountains, but infer only limited expansion from these refugia after the last glaciation. To better understand the genetic dynamics of a South Carpathian spruce lineage, we compared ancient DNA from 10,700 and 11,000-year-old spruce pollen and macrofossils retrieved from Holocene lake sediment in the Retezat Mountains with DNA extracted from extant material from the same site. We used eight primer pairs that amplified short and variable regions of the spruce cpDNA. In addition, from the same lake sediment we obtained a 15,000-years-long pollen accumulation rate (PAR) record for spruce that helped us to infer changes in population size at this site. RESULTS: We obtained successful amplifications for Norway spruce from 17 out of 462 pollen grains tested, while the macrofossil material provided 22 DNA sequences. Two fossil sequences were found to be unique to the ancient material. Population genetic statistics showed higher genetic diversity in the ancient individuals compared to the extant ones. Similarly, statistically significant Ks and Kst values showed a considerable level of differentiation between extant and ancient populations at the same loci.Lateglacial and Holocene PAR values suggested that population size of the ancient population was small, in the range of 1/10 or 1/5 of the extant population. PAR analysis also detected two periods of rapid population growths (from ca. 11,100 and 3900 calibrated years before present (cal yr BP)) and three bottlenecks (around 9180, 7200 and 2200 cal yr BP), likely triggered by climatic change and human impact. CONCLUSION: Our results suggest that the paternal lineages observed today in the Retezat Mountains persisted at this site at least since the early Holocene. Combination of the results from the genetic and the PAR analyses furthermore suggests that the higher level of genetic variation found in the ancient populations and the loss of ancient allele types detected in the extant individuals were likely due to the repeated bottlenecks during the Holocene; however our limited sample size did not allow us to exclude sampling effect.This study demonstrates how past population size changes inferred from PAR records can be efficiently used in combination with ancient DNA studies. The joint application of palaeoecological and population genetics analyses proved to be a powerful tool to understand the influence of past population demographic changes on the haplotype diversity and genetic composition of forest tree species.