Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae.

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N',N″-triacetyl-ß-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.

Influence of Different Diets on Growth and Nutritional Composition of Yellow Mealworm.

Insects are a pathway through which agro-food waste can become a high-quality source of nutrients for both livestock and humans. Yellow mealworm (Tenebrio molitor L., Coleoptera: Tenebrionidae) larvae are currently one of the insect species permitted for consumption, and they are reared on a large scale in Europe. This study evaluated the effect of seven diets containing byproducts such as wheat, rye bran, rapeseed meal, rapeseed cake, flax, and milk thistle cakes on the insect's growth performance, feed conversion ratio (FCR), efficiency of conversion of ingested feed (ECI), nutritional quality of the larvae, and the composition of fats and fatty acids they contained. The lowest FCR based on the fresh and dry basis was 3.32 and 2.01, respectively. The ECI values were statistically different for larvae reared on different experimental diets (mean value 45.7%). As for the nutritional profile, protein and fats ranged from 43.6 to 53.4% d.m. and from 22.3 to 30.0% d.m., respectively. The major fatty acids in all samples were oleic acid (32.97-46.74% of total fatty acids (TFA)), linoleic acid (22.79-38.98% of TFA), and palmitic acid (12.80-17.81% of TFA). This study offers a new opportunity to use and efficiently convert cheap industrial byproducts using yellow mealworms.

Bioconversion Potential of Agro-Industrial Byproducts by Tenebrio molitor-Long-Term Results.

The aim of this study was to compare the growth performance, feed conversion ratio (FCR), and efficiency of ingested feed (ECI) by larvae of the Tenebrio molitor beetle. The growth and development potential of the yellow mealworm was evaluated by using chicken feed (CF), rapeseed meal (RM), wheat bran (WB), and willowleaf sunflower (WS) to obtain inexpensive and various sources of feed. The results showed that the fastest growth with shorter larval development time (74.7 days) was found for insects fed a WB 100 diet. Regarding the final fresh weight of larvae, the highest values were found for larvae grown on WB diets. However, the lowest FCR (1.53 to 1.59) was found for larvae fed RM diets mixed with CF and on the control diet (CF 100). In most cases, it was found that byproduct diets mixed with CF improve the ECI of mealworms, thus contributing to an efficient bioconversion of byproducts into edible sources of nutrients. Thus, except for pure WS as a feed, these byproducts can be used to grow yellow mealworm larvae and may be added to the versatile list of substrates and feed used by small- and large-scale insect producers.

The Future of Climate-Resilient and Climate-Neutral City in the Temperate Climate Zone.

The urban heat island (UHI) effect is the main problem regarding a city's climate. It is the main adverse effect of urbanization and negatively affects human thermal comfort levels as defined by physiological equivalent temperature (PET) in the urban environment. Blue and green infrastructure (BGI) solutions may mitigate the UHI effect. First, however, it is necessary to understand the problem from the degrading side. The subject of this review is to identify the most essential geometrical, morphological, and topographical parameters of the urbanized environment (UE) and to understand the synergistic relationships between city and nature. A four-stage normative procedure was used, appropriate for systematic reviews of the UHI. First, one climate zone (temperate climate zone C) was limited to unify the design guidelines. As a result of delimitation, 313 scientific articles were obtained (546 rejected). Second, the canonical correlation analysis (CCA) was performed for the obtained data. Finally, our research showed the parameters of the UE facilities, which are necessary to mitigate the UHI effect. Those are building density and urban surface albedo for neighborhood cluster (NH), and distance from the city center, aspect ratio, ground surface albedo, and street orientation for street canyon (SC), as well as building height, material albedo, and building orientation for the building structure (BU). The developed guidelines can form the basis for microclimate design in a temperate climate. The data obtained from the statistical analysis will be used to create the blue-green infrastructure (BGI) dynamic modeling algorithm, which is the main focus of the future series of articles.

Antimicrobial Activities Evaluation and Phytochemical Screening of Some Selected Plant Materials Used in Traditional Medicine.

Plant extracts are a source of valuable ingredients that can be used in many industries. This paper presents research on the content of selected bioactive compounds in extracts obtained from various plant materials. Raw materials have a documented use in traditional medicine not only in Poland. The tested plants were: bitter melon (fruit), elderberry (flowers, fruit, leaves), wild rose (fruit, flesh, seeds), mountain ash (fruit), guelder rose (fruit), and sea buckthorn (fruit, leaves, pomace). The main goal of these tests is to indicate the potential raw materials that may constitute an alternative source of bioactive compounds with antimicrobial activity. The plant material was tested for the content of bioactive antioxidant compounds and possible antimicrobial activity. The content of polyphenols (phenolic acids and flavonoids) was analyzed using UPLC/PDA, sterols, organic acids, and other bioactive compounds. The minimum inhibitory concentration (MIC) was determined. The total free phenolic acids (TPC) and total free flavonoids (TFC) of all plant raw materials was varied and ranged from 0.21 (mg RUTE/1 g of extract) to 38.30 mg RUTE/1 g of extract) for TFC. The concentration of sterols was, on average, about 10 mg/1 g of extract. The value of approx. 20 mg/1 g of the extract was recorded for bitter melon and beach rose. The content of organic acids was about 1.5 mg/1 g of the extract to even 13 mg/1 g of the extract for sea buckthorn berries. The most sensitive to the extracts' activity were the following bacteria: M. luteus, P. mirabilis, P. fragii, S. enteritidis, and E. coli. The tested plant materials can be used in various industries as a source of bioactive compounds of an antibacterial nature.

Statistical Review of Quality Parameters of Blue-Green Infrastructure Elements Important in Mitigating the Effect of the Urban Heat Island in the Temperate Climate (C) Zone.

Urban Heat Island (UHI) effect relates to the occurrence of a positive heat balance, compared to suburban and extra-urban areas in a high degree of urbanized cities. It is necessary to develop effective UHI prevention and mitigation strategies, one of which is blue-green infrastructure (BGI). Most research work comparing impact of BGI parameters on UHI mitigation is based on data measured in different climate zones. This makes the implication of nature-based solutions difficult in cities with different climate zones due to the differences in the vegetation time of plants. The aim of our research was to select the most statistically significant quality parameters of BGI elements in terms of preventing UHI. The normative four-step data delimitation procedure in systematic reviews related to UHI literature was used, and temperate climate (C) zone was determined as the UHI crisis area. As a result of delimitation, 173 publications qualified for literature review were obtained (488 rejected). We prepared a detailed literature data analysis and the CVA model-a canonical variation of Fisher's linear discriminant analysis (LDA). Our research has indicated that the BGI object parameters are essential for UHI mitigation, which are the following: area of water objects and green areas, street greenery leaf size (LAI), green roofs hydration degree, and green walls location. Data obtained from the statistical analysis will be used to create the dynamic BGI modeling algorithm, which is the main goal of the series of articles in the future.

How does the content of nutrients in soil affect the health status of trees in city parks?

Trees have multi-aspect influence on the microclimate in urbanised areas. Therefore, it is important to investigate the biotic and abiotic factors affecting their health. The aim of the conducted study was to assess the chemical composition of soils and the nutritional status of lime and horse chestnut trees in selected sites and the influence of these factors on the condition and health of these tree species in urbanised areas. The research was conducted on selected trees (n = 643) growing in different parts of the city. The soils and plants were analysed for the content of macro- and microelements, sodium and heavy metals. A canonical variation analysis (CVA)-the canonical variant of Fisher's linear discriminant analysis (LDA) was used to construct the model. The CVA enabled the creation of 4 CCA models. The research showed that in general, the soil in all the sites of lime and horse chestnut trees was alkalised-at the same time it was characterised by low salinity. Despite the alkaline soil the statistical analysis showed a positive correlation between the content of manganese in the lime leaves and the deterioration of their health. In spite of that due to the satisfactory health status and condition of trees in most locations temporary guide values of nutrients were proposed for trees growing in urbanised areas. The following temporary guide values of nutrients were proposed for the horse chestnut trees (% d. m.): N 2.38%-4.71%, P 0.24%-0.46%, K 1.13%-2.31%, Ca 1.05%-2.12%, Mg 0.16%-0.42%, S 0.12%-0.23%; Fe 89.8-198.8, Zn 17.6-33.1, Cu 7.36-19.61 (mg kg-1 d. m.). The following temporary guide values were proposed for the small-leaved lime-trees (% d. m.): N 2.45%-3.22%, P 0.27%-0.42%, K 1.52%-2.86%, Ca 1.43%-2.02%, Mg 0.19%-0.35%, S 0.19%-0.25%; Fe 137.6-174.3, Zn 20.2-23.8, Cu 8.36-9.79 (mg kg-1 d. m.).