Development and evaluation of SCAR markers for a Pseudomonas brassicacearum strain used in biological control of snow mould

Biological control microorganisms have long been promoted as an alternative to conventional pesticides. Before registration of a microbial biocontrol product for commercial sale, it must be evaluated as regards potential spread and persistence after release. In this study, strainspecific sequence-characterized amplified region (SCAR) markers were developed to monitor the biocontrol candidate strain Pseudomonas brassicacearum MA250, which is effective against snow mould (Microdochium nivale). One SCAR marker, OPA2-73, was used in quantitative real-time PCR (Q-PCR) on samples from a climate chamber experiment in which winter wheat seeds were treated with the bacterium or a chemical control agent, or left untreated. The results showed that MA250 persisted for up to 3 weeks after sowing on the kernel residues and also colonized the roots of treated seedlings. Total MA250 cell numbers on biocontrol treated seedlings after three weeks were approximately 10⁶ cells, compared with the original inoculum of 10⁶–10⁷ cells per seed. Corresponding cell numbers of MA250 on chemically treated and untreated seedlings were below the detection limit. This study shows that SCAR marker OPA2-73 is a specific and sensitive tool for monitoring the biocontrol microorganism MA250 in environmental samples.     

Cytochrome P450 1A, 1B, and 1C mRNA induction patterns in three-spined stickleback exposed to a transient and a persistent inducer

Cytochrome P450 1 (CYP1) mRNA induction patterns in three-spined stickleback (Gasterosteus aculeatus) were explored for use in environmental monitoring of aryl hydrocarbon receptor (AHR) agonists. The cDNAs of stickleback CYP1A, CYP1B1, CYP1C1, and CYP1C2 were cloned and their basal and induced expression patterns were determined in the brain, gill, liver and kidney. Also, their induction time courses were compared after waterborne exposure to a transient (indigo) or a persistent (3,3',4,4',5-pentacholorbiphenyl PCB 126) AHR agonist. The cloned stickleback CYP1s exhibited a high amino acid sequence identity compared with their zebrafish orthologs and their constitutive tissue distribution patterns largely agreed with those reported in other species. PCB 126 (100 nM) induced different CYP1 expression patterns in the four tissues, suggesting tissue-specific regulation. Both indigo (1 nM) and PCB 126 (10 nM) induced a strong CYP1 expression in gills. However, while PCB 126 gave rise to a high and persistent induction in gills and liver, induction by indigo was transient in both organs. The number of putative dioxin response elements found in each CYP1 gene promoter roughly reflected the induction levels of the genes. The high responsiveness of CYP1A, CYP1B1, and CYP1C1 observed in several organs suggests that three-spined stickleback is suitable for monitoring of pollution with AHR agonists.     

Increased Sensitivity to Thermal Pain Following a Single Opiate Dose Is Influenced by the COMT val158met Polymorphism

Increased pain sensitivity after opioid administration (opioid-induced hyperalgesia) and/or repeated painful stimuli is an individually varying and clinically important phenomenon. The functional polymorphism (val¹⁵⁸met) of the Catechol-O-methyltransferase (COMT) gene regulates the metabolism of dopamine/noradrenaline. Individuals homozygous for the met¹⁵⁸ allele have been reported to have increased pain sensitivity and there are findings of lower µ-opioid system activation during sustained pain. We hypothesized that met/met individuals would exhibit higher pain sensitization and opioid-induced hyperalgesia in response to repeated pain stimuli and an intravenous injection of an opioid drug.Participants were 43 healthy subjects who went through an experiment where five blocks of pain were induced to the hand using a heat probe. After each stimulus subjects rated the pain on a visual analogue scale (VAS) from 0 mm (no pain) to 100 mm (worst possible pain). Before the second stimulus there was an intravenous injection of a rapid and potent opioid drug.At baseline there was no difference in pain ratings between the COMTval¹⁵⁸met genotypes, F(2, 39)<1. However, a repeated measures ANOVA for all five stimuli revealed a main effect for COMTval¹⁵⁸met genotype, F(2, 36) = 4.17, p = 0.024. Met/met individuals reported significantly more pain compared to val/val, p = 0.010. A pairwise comparison of baseline and the opioid intervention demonstrated that analgesia was induced in all groups (p = 0.042) without a separating effect for genotype (n.s).We suggest that the initial response of the descending pain system is not influenced by the COMTval¹⁵⁸met polymorphism but when the system is challenged the difference is revealed. An important clinical implication of this may be that the COMTval¹⁵⁸met related differences may be more expressed in individuals where the inhibitory system is already challenged and sensitive, e.g. chronic pain patients. This has to be proven in future studies where the impact of the COMTval¹⁵⁸met polymorphism on opioid treatment in patients is addressed.     

Development and validity of a universal empirical equation to predict skin surface temperature on thermal manikins

Clothing evaporative resistance is an important input in thermal comfort models. Thermal manikin tests give the most accurate and reliable evaporative resistance values for clothing. The calculation methods of clothing evaporative resistance require the sweating skin surface temperature (i.e., options 1 and 2). However, prevailing calculation methods of clothing evaporative resistance (i.e., options 3 and 4) are based on the controlled nude manikin surface temperature due to the sensory measurement difficulty. In order to overcome the difficulty of attaching temperature sensors to the wet skin surface and to enhance the calculation accuracy on evaporative resistance, we conducted an intensive skin study on a thermal manikin ‘Tore’. The relationship among the nude manikin surface temperature, the total heat loss and the wet skin surface temperature in three ambient conditions was investigated. A universal empirical equation to predict the wet skin surface temperature of a sweating thermal manikin was developed and validated on the manikin dressed in six different clothing ensembles. The skin surface temperature prediction equation in an ambient temperature range between 25.0 and 34.0 °C is T_sk=34.0–0.0132HL. It is demonstrated that the universal empirical equation is a good alternative to predicting the wet skin surface temperature and facilitates calculating the evaporative resistance of permeable clothing ensembles. Further studies on the validation of the empirical equation on different thermal manikins are needed however.     

Mental Health Services and Public Safety: Substance Abuse Outpatient Visits Were Associated with Reduced Crime Rates in a Swedish Cohort

Substance abuse is related to offending and substance abuse treatment has been associated with reductions in criminal behavior. This cohort study aimed to explore the relationship between participation in substance abuse interventions and general criminal recidivism among offenders with a combination of mental health problems and substance use problems. In total, 150 Swedish offenders with self-reported mental health and substance use problems were followed for approximately three years with regard to participation in substance abuse interventions and criminal recidivism. Participants with at least three planned visits to specialized outpatient substance abuse clinics had a substantially reduced risk of reoffending as compared to those with fewer than three such visits (HR = 0.47, 95% CI 0.29–0.77). For those with at least three planned visits, general criminal recidivism was reduced by 75% during periods of participation in outpatient visits, as compared to periods of non-participation (HR = 0.25, 95% CI 0.11–0.60). For offenders with mental health problems and substance use problems, outpatient substance abuse interventions could be regarded as important from a clinical risk management perspective, and be encouraged.     

Effects of glucose overloading on microbial community structure and biogas production in a laboratory-scale anaerobic digester

This study characterizes the response of the microbial communities of a laboratory-scale mesophilic biogas process, fed with a synthetic substrate based on cellulose and egg albumin, to single pulses of glucose overloading (15 or 25 times the daily feed based on VS). The microbial biomass and community structure were determined from analyses of membrane phospholipids. The ratio between phospholipid fatty acids (PLFAs; eubacteria and eucaryotes) and di-ethers (PLEL; archaea) suggested that methanogens constituted 4-8% of the microbial biomass. The glucose addition resulted in transient increases in the total biomass of eubacteria while there were only small changes in community structure. The total gas production rate increased, while the relative methane content of the biogas and the alkalinity decreased. However, the biomass of methanogens was not affected by the glucose addition. The results show that the microbial communities of biogas processes can respond quickly to changes in the feeding rate. The glucose overload resulted in a transient general stimulation of degradation rates and almost a doubling of eubacterial biomass, although the biomass increase corresponded to only 7% of the glucose C added.     

Elevated [CO2] and growth temperature have a small positive effect on photosynthetic thermotolerance of Pinus taeda seedlings

Key message

Growth temperature had little effect on the response of net photosynthesis to high temperatures (up to 47 °C). On the other hand, elevated [CO ₂ ] increased net photosynthesis at high temperatures.

Abstract

We investigated whether Pinus taeda seedlings grown under elevated CO₂-concentration ([CO₂]) and temperature would be able to maintain positive net photosynthesis (A _net) longer than seedlings grown under ambient conditions when exposed to temperatures up to 47 °C. Additionally, we investigated whether a locally applied temperature increase would yield the same short-term gas exchange response to temperatures up to 47 °C as a naturally occurring latitudinal temperature increase of equal magnitude. Growth conditions were applied for 7 months (February to August) in treatment chambers constructed at two sites in the native range of P. taeda in the southern US. The sites were located 300 km apart along a north–south axis with a natural temperature difference of 2.1 °C. Seedlings were grown under ambient temperature and [CO₂] conditions at both sites. At the northern site, we also applied a temperature increase of 2 °C (T _E), ensuring that this treatment equalled the mean temperature at the southern site. Additionally, at the northern site, we applied a treatment of elevated [CO₂] (C _E). Gas exchange was measured on all plants in walk-in environmentally controlled chambers. Under C _E, there was no difference in A _net of seedlings grown in ambient or ambient +2 °C temperatures at any measurement temperature, while differences were present under ambient [CO₂]. Furthermore, A _net was higher under C _E than under ambient [CO₂]. At 47 °C, A _net was negative in all seedlings except those in the C _E and ambient temperature treatment combination. Seedlings at the northern site in the T _E treatment showed no significant differences in A _net compared with seedlings grown at ambient temperature at the southern site, indicating that the plants responded equally to a manipulated temperature increase and a latitudinal increase in temperature. Our results suggest that elevated [CO₂] increases photosynthetic thermotolerance at high temperature (>41 °C), but this effect diminishes as temperature increases further. Temperature manipulations could provide accurate information on the effect of latitudinal differences in temperature on leaf gas exchange of P. taeda.     

Cage Culture Turbidostat: a Device for Rapid Determination of Algal Growth Rate

The present cage culture turbidostat consists of a growth chamber and a control unit. The microorganisms (photoautotrophic algae) are kept in the growth chamber by porous membranes (pore size 1 to 3 μm) which retain the algae but allow efficient exchange of the growth medium. Flow rate and composition of the medium can therefore be varied independently of algal population density. A reciprocating pumping mode of the medium is introduced to obtain more gentle clearance of membranes than that provided by rotation or stirring in other membrane fermentors. Pulsed light and a light-emitting diode/light-sensitive transistor couple are used to monitor the turbidity of the culture, independent of external light needed for growth. The control unit keeps the turbidity constant by frequent activation of the dilution pump. Theoretical analysis of growth in the turbidostat shows that integrated activation time of the dilution pump is proportional to the growth rate of the organism. Theoretical analysis was also used to determine minimum flow-rate and nutrient concentration of medium to cover the requirement of the algae. Experiments with three different marine diatoms were carried out, and they demonstrated that the growth rate could be determined every hour and that the cultures could be kept at constant turbidity over 10 to 14 days at least.