Combined effects of three insecticides with different modes of action on biochemical responses of the solitary bee Osmia bicornis.

Bees are simultaneously exposed to a variety of pesticides, which are often applied in mixtures and can cause lethal and sublethal effects. The combined effects of pesticides, however, are not measured in the current risk assessment schemes. Additionally, the sublethal effects of pesticides on a variety of physiological processes are poorly recognized in bees, especially in non-Apis solitary bees. In this study, we used a full-factorial design to examine the main and interactive effects of three insecticide formulations with different modes of action (Mospilan 20 SP, Sherpa 100 EC, and Dursban 480 EC) on bee biochemical processes. We measured acetylcholinesterase (AChE), glutathione S-transferase (GST) and esterase (EST) activities, as well as a nonenzymatic biomarker associated with energy metabolism, i.e., ATP level. All studied endpoints were affected by Sherpa 100 EC, and the activities of AChE and EST as well as ATP levels were affected by Dursban 480 EC. Moreover, complex interactions between all three insecticides affected ATP levels, showing outcomes that cannot be predicted when testing each insecticide separately. The results indicate that even if interactive effects are sometimes difficult to interpret, there is a need to study such interactions if laboratory-generated toxicity data are to be extrapolated to field conditions.

Floral resources,energetic value and pesticide residues in provisions collected by Osmia bicornis along a gradient of oilseed rape coverage.

Pollinators in agricultural landscapes are facing global decline and the main pressures include food scarcity and pesticide usage. Intensive agricultural landscapes may provide important food resources for wild pollinators via mass flowering crops. However, these are monofloral, short-term, and may contain pesticide residues. We explored how the landscape composition with a different proportion of oilseed rape (6-65%) around Osmia bicornis nests affects floral diversity, contamination with pesticides, and energetic value of provisions collected by this species of wild bees as food for their offspring. Altogether, the bees collected pollen from 28 plant taxa (6-15 per nest) and provisions were dominated by Brassica napus (6.0-54.2%, median 44.4%, 12 nests), Quercus sp. (1.2-19.4%, median 5.2%, 12 nests), Ranunculus sp. (0.4-42.7%, median 4.7%, 12 nests), Poaceae (1.2-59.9%, median 5.8%, 11 nests) and Acer sp. (0.6-42%, median 18.0%, 8 nests). Residues of 12 pesticides were found in provisions, with acetamiprid, azoxystrobin, boscalid, and dimethoate being the most frequently detected at concentrations up to 1.2, 198.4, 16.9 and 17.8 ng/g (median 0.3, 10.6, 11.3, 4.4 ng/g), respectively. Floral diversity and energetic value of provisions, but not the Pesticide Risk Index depended on landscape structure. Moreover, pollen diversity decreased, and energetic value increased with landscape diversity. Thus, even a structurally simple landscape may provide diverse food for O. bicornis if the nest is located close to a single but resource-diverse patch. Both B. napus and non-crop pollen were correlated with pesticide concentrations.

Congruence between the prioritisation of conservation problems at the local and national scale: an evaluation by environmental scientists in Poland.

The anthropogenic pressure on the environment depends on the spatial scale. It is crucial to prioritise conservation actions at different spatial scales to be cost-efficient. Using horizon scanning with the Delphi technique, we asked what the most important conservation problems are in Poland at local and national scales. Twenty-six participants, PhD students, individually identified conservation issues important at the local and national scales. Each problem was then scored and classified into broader categories during the round discussions. Text mining, cross-sectional analyses, and frequency tests were used to compare the context, importance scores, and frequency of identified problems between the two scales, respectively. A total of 115 problems were identified at the local scale and 122 at the national scale. Among them, 30 problems were identical for both scales. Importance scores were higher for national than local problems; however, this resulted from different sets of problems identified at the two scales. Problems linked to urbanisation, education, and management were associated with the local scale. Problems related to policy, forestry, and consumerism were more frequent at the national scale. An efficient conservation policy should be built hierarchically (e.g. introducing adaptive governance), implementing solutions at a national scale with the flexibility to adjust for local differences and to address the most pressing issues.

Bird feathers as potential sources of pathogenic microorganisms: a new look at old diseases.

This article describes methods of treatment for avian zoonoses, modern antibiotic therapy and drug resistance of selected pathogens, which pose a threat to the population's health. A tabular form has been used to present the current data from the European Union from 2011 to 2017 regarding human morbidity and mortality and the costs incurred by national health systems for the treatment of zoonoses occurring in humans and animals. Moreover, the paper includes descriptions of selected diseases, which indirectly affect birds. Scientists can obtain information regarding the occurrence of particular diseases, their aetiology, epidemiology, incubation period and symptoms caused by dangerous microorganisms and parasites. This information should be of particular interest for people who have frequent contact with birds, such as ornithologists, as well as veterinarians, farm staff, owners of accompanying animals and zoological workers. This paper presents a review used for identification and genetic characterization of bacterial strains isolated from a variety of environmental sources, e.g., bird feathers along with their practical application. We describe the bacterial, viral and fungal serotypes present on avian feathers after the slaughter process. This review also enables us to effectively identify several of the early stages of infectious diseases from heterogeneous avian research material.

Expression of GPD1 and SIP18 genes during rehydration in active dry industrial Saccharomyces cerevisiae cider-making yeast strains (ADY).

In this study we determined the influence of different sugar concentration in media, time of rehydration and type of strain on relative expression level of GPD1 and SIP18 genes of active dry cider-making yeast strains, followed by the assessment of the impact of rehydration on the fermentation process. High expression of SIP18 at the beginning of rehydration was shown to be due to high transcription of the gene during the drying process. High sugar concentrations of media initiated transcription of the GPD1 gene and triggered the cellular glycerol biosynthesis pathway in examined strains. Rehydration time and type of strain showed to have no statistically significant impact on the course of the fermentation; RT qPCR results depended mainly on the time of rehydration and sugar concentration of the medium. This is the first attempt to confront rehydration time and molecular mechanisms acting upon rehydration with the course of the fermentation process.

The Effect of a Newly Developed Oat-Banana Fermented Beverage with a Beta-glucan Additive on ldhL Gene Expression in Streptococcus thermophilus TKM3 KKP 2030p.

The number of people who cannot consume dairy products due to intolerance or allergies to food components is increasing. Consumers are increasingly searching for alternative, nondairy beverages which have an advantageous effect on the body and which stimulate gut microflora. Previous studies have shown that Streptococcus thermophilus TKM3 KKP 2030p can be an efficient starter culture for the fermentation of plant material and can also ensure an acceptable sensory profile and a high number of lactic acid bacteria during 4 week cold storage. The aim of this study was to determine the relative gene expression of the L-lactate dehydrogenase gene (ldhL) in selected strain using the RT qPCR technique at particular time points. The gene expression experiments were conducted in laboratory broth (LABm) and fruit-cereal (OBPromOat) matrix with a beta-glucan additive. Levels of gene relative expression in the selected strain in these two media were correlated with the amount of L-lactate produced. The results showed that the plant matrix stimulated stronger ldhL gene expression in the first 4 h of the experiment (although the synthesis of L-lactate was less than in laboratory broth). This study broadens existing knowledge of the transcriptional mechanisms which arise from the adaptation of the analyzed strain to different habitats during L-lactate synthesis. This could contribute to the development of new alternative food products for people who show intolerance to milk proteins or lactose.

Intra-strains diversity of expression of polymorphic PKS4 gene in comparison in zearalenone production by Fusarium graminearum during in vitro cultivation.

Filamentous fungi belonging to the Fusarium genus are responsible for large economic losses due to their high pathogenicity and toxigenicity. Fusarium sp. may produce variety of mycotoxins, one of them is zearalenone (ZEA). The presence of the PKS4 gene shows the possibility of zearalenone biosynthesis by Fusarium sp. In this study, in four Fusarium graminearum and one Fusarium poae strains the presence of PKS4 genes and ZEA concentrations were determined. The presence of the PKS4 gene was confirmed by classical polymerase chain reaction (PCR) in three of four strains of F. graminearum. One strain with no PKS4 gene detected was found while still producing ZEA. In the present study, a real-time PCR assay has been successfully performed for the relative expression of Fusarium strains based on new designed primers targeting the PKS4 gene involved in ZEA biosynthesis. Result shows that P56/4 strain of F. graminearum has the highest mRNA level, in the range of 12, what correlates to the high production of this mycotoxin. In this study, a real-time PCR assay has been successfully developed for the prediction of the production of ZEA by F. graminearum strains by PCR real-time techniques based on primers targeting the gene, PKS4, involved in ZEA biosynthesis. The special significance was pointed to occurring genes polymorphism.

The sequence diversity and expression among genes of the folic acid biosynthesis pathway in industrial Saccharomyces strains.

Folic acid is an important vitamin in human nutrition and its deficiency in pregnant women's diets results in neural tube defects and other neurological damage to the fetus. Additionally, DNA synthesis, cell division and intestinal absorption are inhibited in case of adults. Since this discovery, governments and health organizations worldwide have made recommendations concerning folic acid supplementation of food for women planning to become pregnant. In many countries this has led to the introduction of fortifications, where synthetic folic acid is added to flour. It is known that Saccharomyces strains (brewing and bakers' yeast) are one of the main producers of folic acid and they can be used as a natural source of this vitamin. Proper selection of the most efficient strains may enhance the folate content in bread, fermented vegetables, dairy products and beer by 100% and may be used in the food industry. The objective of this study was to select the optimal producing yeast strain by determining the differences in nucleotide sequences in the FOL2, FOL3 and DFR1 genes of folic acid biosynthesis pathway. The Multitemperature Single Strand Conformation Polymorphism (MSSCP) method and further nucleotide sequencing for selected strains were applied to indicate SNPs in selected gene fragments. The RT qPCR technique was also applied to examine relative expression of the FOL3 gene. Furthermore, this is the first time ever that industrial yeast strains were analysed regarding genes of the folic acid biosynthesis pathway. It was observed that a correlation exists between the folic acid amount produced by industrial yeast strains and changes in the nucleotide sequence of adequate genes. The most significant changes occur in the DFR1 gene, mostly in the first part, which causes major protein structure modifications in KKP 232, KKP 222 and KKP 277 strains. Our study shows that the large amount of SNP contributes to impairment of the selected enzymes and S. cerevisiae and S. pastorianus produce reduced amounts of the investigated metabolite. The results obtained here yield a list of genetically stable yeast strains which can be implemented as a starter culture in the food industry.

Microbial transglutaminase and its application in the food industry. A review.

The extremely high costs of manufacturing transglutaminase from animal origin (EC 2.3.2.13) have prompted scientists to search for new sources of this enzyme. Interdisciplinary efforts have been aimed at producing enzymes synthesised by microorganisms which may have a wider scope of use. Transglutaminase is an enzyme that catalyses the formation of isopeptide bonds between proteins. Its cross-linking property is widely used in various processes: to manufacture cheese and other dairy products, in meat processing, to produce edible films and to manufacture bakery products. Transglutaminase has considerable potential to improve the firmness, viscosity, elasticity and water-binding capacity of food products. In 1989, microbial transglutaminase was isolated from Streptoverticillium sp. Its characterisation indicated that this isoform could be extremely useful as a biotechnological tool in the food industry. Currently, enzymatic preparations are used in almost all industrial branches because of their wide variety and low costs associated with their biotechnical production processes. This paper presents an overview of the literature addressing the characteristics and applications of transglutaminase.

Correlation between acetic acid resistance and characteristics of PQQ-dependent ADH in acetic acid bacteria.

In this study, we compared the growth properties and molecular characteristics of pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase (ADH) among highly acetic acid-resistant strains of acetic acid bacteria. Gluconacetobacter europaeus exhibited the highest resistance to acetic acid (10%), whereas Gluconacetobacter intermedius and Acetobacter pasteurianus resisted up to 6% of acetic acid. In media with different concentrations of acetic acid, the maximal acetic acid production rate of Ga. europaeus slowly increased, but specific growth rates decreased concomitant with increased concentration of acetic acid in medium. The lag phase of A. pasteurianus was twice and four times longer in comparison to the lag phases of Ga. europaeus and Ga. intermedius, respectively. PQQ-dependent ADH activity was twice as high in Ga. europaeus and Ga. intermedius as in A. pasteurinus. The purified enzymes showed almost the same specific activity to each other, but in the presence of acetic acid, the enzyme activity decreased faster in A. pasteurianus and Ga. intermedius than in Ga. europaeus. These results suggest that high ADH activity in the Ga. europaeus cells and high acetic acid stability of the purified enzyme represent two of the unique features that enable this species to grow and stay metabolically active at extremely high concentrations of acetic acid.

Intensification of lipase biosynthesis as a result of electrofusion of Rhizopus cohnii protoplasts.

Our objective was to obtain products of fusion of the filamentous fungus Rhizopus cohnii Rh.c./1 with an increased capacity for lipase biosynthesis in comparison with the original strain. Protoplasts of auxotrophic mutants of the parent strain Rh.c./1 obtained after UV irradiation of the spores were subjected to electrofusion. We found that the largest number of electrofusion products could be obtained with the use of the following process parameters: 1 or 2 impulses immediately following one another with a field intensity of 200 V/cm and an exposition time of 1000 ms at the stage of dielectrophoresis, 1 impulse with a field intensity of 500 V/cm and an exposition time of 10 ms or 20 ms at the stage of fusion, regulated temperature of 4 degrees C before and after the process, rounding time of ca 20 min. Electrofusion of protoplasts of auxotrophic mutants of the Rh.c./1 strain produced 19 fusion products whose lipase biosynthesis capacity in a liquid medium culture was higher than that of the parent strains. The fusion product labelled XIII-21 was selected as the best strain. Lipase activity obtained after its culture in the liquid medium was ca 3.5 times higher than that obtained after the culture of the original strain Rh.c./1.