Search

Carbonyl-reducing enzymes (CREs) catalyse the reduction of carbonyl groups in many eobiotic and xenobiotic compounds in all organisms, including helminths. Previous studies have shown the important roles of CREs in the deactivation of several anthelmintic drugs (e.g., flubendazole and mebendazole) in adults infected with the parasitic nematode Haemonchus contortus, in which the activity of a CRE is increased in drug-resistant strains. The aim of the present study was to compare the abilities of nematodes of both a drug-susceptible strain (ISE) and a drug-resistant strain (IRE) to reduce the carbonyl group of flubendazole (FLU) in different developmental stages (eggs, L1/2 larvae, L3 larvae, and adults). In addition, the effects of selected CRE inhibitors (e.g., glycyrrhetinic acid, naringenin, silybin, luteolin, glyceraldehyde, and menadione) on the reduction of FLU were evaluated in vitro and ex vivo in H. contortus adults. The results showed that FLU was reduced by H. contortus in all developmental stages, with adult IRE females being the most metabolically active. Larvae (L1/2 and L3) and adult females of the IRE strain reduced FLU more effectively than those of the ISE strain. Data from the in vitro inhibition study (performed with cytosolic-like fractions of H. contortus adult homogenate) revealed that glycyrrhetinic acid, naringenin, mebendazole and menadione are effective inhibitors of FLU reduction. Ex vivo study data showed that menadione inhibited FLU reduction and also decreased the viability of H. contortus adults to a similar extent. Naringenin and mebendazole were not toxic at the concentrations tested, but they did not inhibit the reduction of FLU in adult worms ex vivo. [ABSTRACT FROM AUTHOR]

Poly-β-hydroxybutyrate (PHB) is a potential source of biodegradable plastics that are environmentally friendly due to their complete degradation to water and carbon dioxide. This study aimed to investigate PHB production in the cyanobacterium Synechocystis sp. PCC6714 MT_a24 in an outdoor bioreactor using urban wastewater as a sole nutrient source. The culture was grown in a thin-layer raceway pond with a working volume of 100 L, reaching a biomass density of up to 3.5 g L−1 of cell dry weight (CDW). The maximum PHB content was found under nutrient-limiting conditions in the late stationary phase, reaching 23.7 ± 2.2% PHB per CDW. These data are one of the highest reported for photosynthetic production of PHB by cyanobacteria, moreover using urban wastewater in pilot-scale cultivation which multiplies the potential of sustainable cultivation approaches. Contamination by grazers (Poterioochromonas malhamensis) was managed by culturing Synechocystis in a highly alkaline environment (pH about 10.5) which did not significantly affect the culture growth. Furthermore, the strain MT_a24 showed significant wastewater nutrient remediation removing about 72% of nitrogen and 67% of phosphorus. These trials demonstrate that the photosynthetic production of PHB by Synechocystis sp. PCC6714 MT_a24 in the outdoor thin-layer bioreactor using urban wastewater and ambient carbon dioxide. It shows a promising approach for the cost-effective and sustainable production of biodegradable carbon-negative plastics. Key points: • High PHB production by cyanobacteria in outdoor raceway pond • Urban wastewater used as a sole source of nutrients for phototrophic growth • Potential for cost-effective and sustainable production of biodegradable plastics [ABSTRACT FROM AUTHOR]

Poly-β-hydroxybutyrate (PHB) is a potential source of biodegradable plastics that are environmentally friendly due to their complete degradation to water and carbon dioxide. This study aimed to investigate PHB production in the cyanobacterium Synechocystis sp. PCC6714 MT_a24 in an outdoor bioreactor using urban wastewater as a sole nutrient source. The culture was grown in a thin-layer raceway pond with a working volume of 100 L, reaching a biomass density of up to 3.5 g L−1 of cell dry weight (CDW). The maximum PHB content was found under nutrient-limiting conditions in the late stationary phase, reaching 23.7 ± 2.2% PHB per CDW. These data are one of the highest reported for photosynthetic production of PHB by cyanobacteria, moreover using urban wastewater in pilot-scale cultivation which multiplies the potential of sustainable cultivation approaches. Contamination by grazers (Poterioochromonas malhamensis) was managed by culturing Synechocystis in a highly alkaline environment (pH about 10.5) which did not significantly affect the culture growth. Furthermore, the strain MT_a24 showed significant wastewater nutrient remediation removing about 72% of nitrogen and 67% of phosphorus. These trials demonstrate that the photosynthetic production of PHB by Synechocystis sp. PCC6714 MT_a24 in the outdoor thin-layer bioreactor using urban wastewater and ambient carbon dioxide. It shows a promising approach for the cost-effective and sustainable production of biodegradable carbon-negative plastics. Key points: • High PHB production by cyanobacteria in outdoor raceway pond • Urban wastewater used as a sole source of nutrients for phototrophic growth • Potential for cost-effective and sustainable production of biodegradable plastics [ABSTRACT FROM AUTHOR]

Microalgae are excellent sources of biomass containing several important compounds for human and animal nutrition—proteins, lipids, polysaccharides, pigments and antioxidants as well as bioactive secondary metabolites. In addition, they have a great biotechnological potential for nutraceuticals, and pharmaceuticals as well as for CO2 sequestration, wastewater treatment, and potentially also biofuel and biopolymer production. In this review, the industrial production of the most frequently used microalgae genera—Arthrospira, Chlorella, Dunaliella, Haematococcus, Nannochloropsis, Phaeodactylum, Porphyridium and several other species is discussed as concerns the applicability of the most widely used large-scale systems, solar bioreactors (SBRs)—open ponds, raceways, cascades, sleeves, columns, flat panels, tubular systems and others. Microalgae culturing is a complex process in which bioreactor operating parameters and physiological variables closely interact. The requirements of the biological system—microalgae culture are crucial to select the suitable type of SBR. When designing a cultivation process, the phototrophic production of microalgae biomass, it is necessary to employ SBRs that are adequately designed, built and operated to satisfy the physiological requirements of the selected microalgae species, considering also local climate. Moreover, scaling up microalgae cultures for commercial production requires qualified staff working out a suitable cultivation regime. Key points: • Large-scale solar bioreactors designed for microalgae culturing. • Most frequently used microalgae genera for commercial production. • Scale-up requires suitable cultivation conditions and well-elaborated protocols. [ABSTRACT FROM AUTHOR]

Photosynthesis, growth and biochemical composition of the biomass of the freshwater microalga Chlamydopodium fusiforme cultures outdoors in a thin-layer cascade were investigated. Gross oxygen production measured off-line in samples taken from the outdoor cultures was correlated with the electron transport rate estimated from chlorophyll a fluorescence measurements. According to photosynthesis measurements, a mean of 38.9 ± 10.3 mol of photons were required to release one mole of O2, which is 4.86 times higher than the theoretical value (8 photons per 1 O2). In contrast, according to the fluorescence measurements, a mean of 11.7 ± 0.74 mol of photons were required to release 1 mol of O2. These findings indicate that fluorescence-based photosynthesis rates may not be fully replace oxygen measurements to evaluate the performance of an outdoor culture. Daily gross biomass productivity was 0.3 g DW L−1 day−1 consistently for 4 days. Biomass productivity was strongly affected by the suboptimal concentration at which the culture was operated and by the respiration rate, as the substantial volume of culture was kept in the dark (about 45% of the total volume). As the cells were exposed to excessive light, the photosynthetic activity was mainly directed to the synthesis of carbohydrates in the biomass. In the morning, carbohydrate content decreased because of the dark respiration. Per contra, protein content in the biomass was lower at the end of the day and higher in the morning due to carbohydrate consumption by respiration. The data gathered in these trials are important for the future exploitation of Chlamydopodium fusiforme as a potential novel species in the field of microalgae for the production of bio-based compounds. [ABSTRACT FROM AUTHOR]

In this work, four microalgae species (Chlorella vulgaris, Trachydiscus minutus, Monodopsis sp., and Monoraphidium sp.) were selected and grown in a novel, 30 L short-light path annular-column photobioreactor (AC-PBR). The aim was to test the microalgae growth on a pilot scale and evaluate the potential use as an alternative fish feed in hatcheries based on the content and composition of fatty acids. All microalgae species reached a high biomass density between 3.65 and 5.32 g DW L−1 during a 14-day trial. The highest growth rate of 0.25 ± 0.05 d−1 was calculated for Trachydiscus culture. The highest total content of fatty acids (TFA) was determined for Monodopsis and Trachydiscus biomass as the concentration reached 74.5 ± 6.3 and 72.4 ± 7.6 mg g−1 DW, respectively. These two species contained mostly eicosapentaenoic acid (EPA; C20:5n3), the crucial FA for carnivorous fish whose concentration reached the value of 25.3 ± 0.9% and 31.9 ± 2.0% of TFA for Monodopsis and Trachydiscus, respectively. A high amount of linoleic acid (LA; C18:2n6), another important FA for freshwater aquafeed, was found in Chlorella biomass up to the concentration of 21.9 ± 1.5% of TFA. The present results suggest the feasibility of in-house cultivation of selected microalgae species as a quality live feed source for fish hatcheries. [ABSTRACT FROM AUTHOR]

The microalga Chlamydopodium fusiforme MACC-430 was cultured in two types of outdoor pilot cultivation units—a thin-layer cascade (TLC) and a raceway pond (RWP) placed in a greenhouse. This case study aimed to test their potential suitability for cultivation scale-up to produce biomass for agriculture purposes (e.g., as biofertilizer or biostimulant). The culture response to the alteration of environmental conditions was evaluated in "exemplary" situations of good and bad weather conditions using several photosynthesis measuring techniques, namely oxygen production, and chlorophyll (Chl) fluorescence. Validation of their suitability for online monitoring in large-scale plants has been one of the objectives of the trials. Both techniques were found fast and robust reliable to monitor microalgae activity in large-scale cultivation units. In both bioreactors, Chlamydopodium cultures grew well in the semi-continuous regime using daily dilution (0.20—0.25 day−1). The biomass productivity calculated per volume was significantly (about 5 times) higher in the RWPs compared to the TLCs. The measured photosynthesis variables showed that the build-up of dissolved oxygen concentration in the TLC was higher, up to 125–150% of saturation (%sat) as compared to the RWP (102–104%sat). As only ambient CO2 was available, its shortage was indicated by a pH increase due to photosynthetic activity in the thin-layer bioreactor at higher irradiance intensities. In this setup, the RWP was considered more suitable for scale-up due to higher areal productivity, lower construction and maintenance costs, the smaller land area required to maintain large culture volumes, as well as lower carbon depletion and dissolved oxygen build-up. Key points: • Chlamydopodium was grown in both raceways and thin-layer cascades in pilot-scale. • Various photosynthesis techniques were validated for growth monitoring. • In general, raceway ponds were evaluated as more suitable for cultivation scale-up. [ABSTRACT FROM AUTHOR]

Short-chain dehydrogenases/reductases (SDRs) regulate the activities of many hormones and other signaling molecules and participate in the deactivation of various carbonyl-bearing xenobiotics. Nevertheless, knowledge about these important enzymes in helminths remains limited. The aim of our study was to characterize the SDR superfamily in the parasitic nematode Haemonchus contortus. Genome localization of SDRs was explored, and phylogenetic analysis in comparison with SDRs from free-living nematode Caenorhabditis elegans and the domestic sheep (Ovis aries, a typical host of H. contortus) was constructed. The expression profile of selected SDRs during the life cycle along with differences between the drug-susceptible and drug-resistant strains, were also studied. Genome sequencing enabled the identification of 46 members of the SDR family in H. contortus. A number of genes have no orthologue in the sheep genome. In all developmental stages of H. contortus, SDR1, SDR3, SDR5, SDR6, SDR14, and SDR18 genes were the most expressed, although in individual stages, huge differences in expression levels were observed. A comparison of SDRs expression between the drug-susceptible and drug-resistant strains of H. contortus revealed several SDRs with changed expression in the resistant strain. Specifically, SDR1, SDR12, SDR13, SDR16 are SDR candidates related to drug-resistance, as the expression of these SDRs is consistently increased in most stages of the drug-resistant H. contortus. These findings revealing several SDR enzymes of H. contortus warrant further investigation. [ABSTRACT FROM AUTHOR]

The nematode Haemonchus contortus , a gastrointestinal parasite of ruminants, can severely burden livestock production. Although anthelmintics are the mainstay in the treatment of haemonchosis, their efficacy diminishes due to drug-resistance development in H. contortus. An increased anthelmintics inactivation via biotransformation belongs to a significant drug-resistance mechanism in H. contortus. UDP-glycosyltransferases (UGTs) participate in the metabolic inactivation of anthelmintics and other xenobiotic substrates through their conjugation with activated sugar, which drives the elimination of the xenobiotics due to enhanced solubility. The UGTs family, in terms of the biotransformation of commonly used anthelmintics, has been well described in adults as a target stage. In contrast, the free-living juvenile stages of H. contortus have attracted less attention. The expression of UGTs considerably varies throughout the life cycle of the juvenile nematodes, suggesting their different roles. Furthermore, the constitutive expression in a susceptible strain with two resistant strains shows several resistance-related changes in UGTs expression, and the exposure of juvenile stages of H. contortus to albendazole (ABZ) and ABZ-sulfoxide (ABZSO; in sublethal concentrations) leads to the increased expression of several UGTs. The anthelmintic drug ABZ and its primary metabolite ABZSO biotransformation, tested in the juvenile stages, shows significant differences between susceptible and resistant strain. Moreover, higher amounts of glycosidated metabolites of ABZ are formed in the resistant strain. Our results show similarly, as in adults, the UGTs and glycosidations significant for resistance-related differences in ABZ biotransformation and warrant further investigation in their individual functions. [ABSTRACT FROM AUTHOR]

The efficacy of anthelmintic therapy of farm animals rapidly decreases due to drug resistance development in helminths. In resistant isolates, the increased expression and activity of drug-metabolizing enzymes (DMEs), e.g. cytochromes P450 (CYPs), UDP-glycosyltransferases (UGTs) and P-glycoprotein transporters (P-gps), in comparison to sensitive isolates have been described. However, the mechanisms and circumstances of DMEs induction are not well known. Therefore, the present study was designed to find the changes in expression of CYPs, UGTs and P-gps in adult parasitic nematodes Haemonchus contortus exposed to sub-lethal doses of the benzimidazole anthelmintic drug albendazole (ABZ) and its active metabolite ABZ-sulfoxide (ABZSO). In addition, the effect of ABZ at sub-lethal doses on the ability to deactivate ABZ during consequent treatment was studied. The results showed that contact of H. contortus adults with sub-lethal doses of ABZ and ABZSO led to a significant induction of several DMEs, particularly cyp-2, cyp-3, cyp-6, cyp-7, cyp-8, UGT10B1, UGT24C1, UGT26A2, UGT365A1, UGT366C1, UGT368B2, UGT367A1, UGT371A1, UGT372A1 and pgp-3, pgp-9.1, pgp-9.2, pgp-10. This induction led to increased formation of ABZ metabolites (especially glycosides) and their increased export from the helminths' body into the medium. The present study demonstrates for the first time that contact of H. contortus with sub-lethal doses of ABZ (e.g. during underdose treatment) improves the ability of H. contortus adults to deactivate ABZ in consequent therapy. [ABSTRACT FROM AUTHOR]

Drug-metabolizing enzymes represent the main defense system against xenobiotics in all organisms. Long-term exposure to drugs can lead to changes in the expression of specific enzymes and to development of drug resistance. The previous studies have shown that increased anthelmintics inactivation via increased expression of certain drugmetabolizing enzymes belongs to a significant mechanism of drug resistance in Haemonchus contortus. H. contortus, a gastrointestinal parasite of ruminants, has developed resistance to all used anthelmintics. As short-chain dehydrogenases/reductases (SDRs) catalyze the deactivation of carbonyl-containing anthelmintics (e.g. flubendazole, mebendazole), the increased expression of these SDRs could promote decreased susceptibility of H. contortus to these anthelmintics. For these reasons, the present study was designed to follow the changes in the relative expression of selected SDR genes in adults of H. contortus exposed to different concentrations of flubendazole (FLU). FLU-mediated responses in adults from drug-susceptible strain (ISE) and drug-resistant strain (IRE) were compared. The adult nematodes were incubated ex vivo with or without FLU (0.01 μM, 0.1 μM, 1 μM, and 5 μM) in a culture medium for 4 h or 12 h. After incubation, total RNA was isolated and the expression level of individual SDRs were analyzed using qPCR. The results have shown the variability in the expression of individual SDR genes in H. contortus adults after contact with sub lethal doses of FLU. [ABSTRACT FROM AUTHOR]