Evaluation of Petrifilm™ EC method for enumeration of E. coli from soil

Aims: To evaluate the suitability of commercially available Petrifilm? EC plates for enumeration of Escherichia coli from soil. Methods and Results: A confirmed E. coli strain isolated from liquid swine manure was inoculated into sterilized sandy clay loam and loam soils at the concentrations of 10², 10³, 10⁵ CFU g^?1 of soil. The efficiency of recovery on Petrifilm? EC plates for soils spiked with E. coli was compared with standard membrane filtration techniques on m‐FC basal medium supplemented with 3‐bromo‐4‐chloro‐5‐indoyl‐β‐d ‐glucopyranoside (BCIG) and most probable numbers (MPN) techniques in E. coli medium with 4‐methylumbelliferyl‐β‐d ‐glucuronide (EC‐MUG) broth. Petrifilm? EC and m‐FC (BCIG) methods were then assessed for the ability to recover E. coli from field soils applied with swine manure. No significant differences (P > 0·05) were observed between Petrifilm? EC, m‐FC (BCIG) and MPN methods for the recovery of E. coli from spiked samples, irrespective of soil type. However, recovery of E. coli from manure‐applied field soil samples showed a significant difference (P < 0·05) between the Petrifilm? EC method and the m‐FC method in enumerating E. coli possibly as a result of false positives on m‐FC. Conclusion: The Petrifilm? EC method is suitable for the enumeration of E. coli from soil with a detection limit of 10 CFU g^?1 soil. Significance and Impact of the Study: The commercially available Petrifilm? EC method is comparatively low cost, easy to use method for the enumeration of E. coli from soil without the need for further confirmation tests.     

Energetics of Acidianus ambivalens growth in response to oxygen availability

All life requires energy to drive metabolic reactions such as growth and cell maintenance; therefore, fluctuations in energy availability can alter microbial activity. There is a gap in our knowledge concerning how energy availability affects the growth of extreme chemolithoautotrophs. Toward this end, we investigated the growth of thermoacidophile Acidianus ambivalens during sulfur oxidation under aerobic to microaerophilic conditions. Calorimetry was used to measure enthalpy (ΔH_inc) of microbial activity, and chemical changes in growth media were measured to calculate Gibbs energy change (ΔG_inc) during incubation. In all experiments, Gibbs energy was primarily dissipated through the release of heat, which suggests enthalpy‐driven growth. In microaerophilic conditions, growth was significantly more efficient in terms of biomass yield (defined as C‐mol biomass per mole sulfur consumed) and resulted in lower ΔG_inc and ΔH_inc. ΔG_inc in oxygen‐limited (OL) and oxygen‐ and CO₂‐limited (OCL) microaerophilic growth conditions resulted in averages of ?1.44 × 10³ kJ/C‐mol and ?7.56 × 10² kJ/C‐mol, respectively, and average ΔH_inc values of ?1.11 × 10⁵ kJ/C‐mol and ?4.43 × 10⁴ kJ/C‐mol, respectively. High‐oxygen experiments resulted in lower biomass yield values, an increase in ΔG_inc to ?1.71 × 10⁴ kJ/C‐mol, and more exothermic ΔH_inc values of ?4.71 × 10⁵ kJ/C‐mol. The observed inefficiency in high‐oxygen conditions may suggest larger maintenance energy demands due to oxidative stresses and a preference for growth in microaerophilic environments.     

Changes in behavior are unable to disrupt a trophic cascade involving a specialist herbivore and its food plant

Changes in ecological conditions can induce changes in behavior and demography of wild organisms, which in turn may influence population dynamics. Black brant (Branta bernicla nigricans) nesting in colonies on the Yukon–Kuskokwim Delta (YKD) in western Alaska have declined substantially (~50%) since the turn of the century. Black brant are herbivores that rely heavily on Carex subspathacea (Hoppner's sedge) during growth and development. The availability of C. subspathacea affects gosling growth rates, which subsequently affect pre‐ and postfledging survival, as well as size and breeding probability as an adult. We predicted that long‐term declines in C. subspathacea have affected gosling growth rates, despite the potential of behavior to buffer changes in food availability during brood rearing. We used Bayesian hierarchical mixed‐effects models to examine long‐term (1987–2015) shifts in brant behavior during brood rearing, forage availability, and gosling growth rates at the Tutakoke River colony. We showed that locomotion behaviors have increased (β = 0.05, 95% CRI: 0.032–0.068) while resting behaviors have decreased (β = ?0.024, 95% CRI: ?0.041 to ?0.007), potentially in response to long‐term shifts in forage availability and brood density. Concurrently, gosling growth rates have decreased substantially (β = ?0.100, 95% CRI: ?0.191 to ?0.016) despite shifts in behavior, mirroring long‐term declines in the abundance of C. subspathacea (β = ?0.191, 95% CRI: ?0.355 to ?0.032). These results have important implications for individual fitness and population viability, where shifts in gosling behavior putatively fail to mitigate long‐term declines in forage availability.     

RPW8 promoter is involved in pathogen‐And stress‐inducible expression from Vitis pseudoreticulata

Based on previous cloning of VpRPW8‐e, we obtained a 1,126 bp VpRPW8‐e promoter sequence in this study. A large number of TATA‐boxes, CAAT‐boxes, and other cis‐acting elements were predicted including light‐responsive elements, hormone‐responsive elements, stress‐responsive elements, and growth‐ and development‐associated elements within the promoter sequence. To further investigate the function of this promoter, we examined its activity in response to biotic and abiotic stress. The VpRPW8‐e promoter was strongly activated by Plasmopara viticola infection, and activation also occurred when the orientation of the promoter was reversed, although to a lesser extent. Deletion analysis showed that the ?1,126 to ?475 bp region of VpRPW8‐e promoter had high activity. A promoter fragment 5′ deleted to ?475 bp (P_?475) was activated in response to heat and cold stress, and even more strongly in response to Phytophthora capsici and salicylic acid (SA). Furthermore, Transgenic Nicotiana benthamiana were generated, VpRPW8‐e driven by P_?475 enhanced resistance to Ph. capsici in N. benthamiana. Based on these results, the ?475 bp region was deduced to be an indispensable part of the VpRPW8‐e promoter. VpRPW8‐e promoter is involved in pathogen‐ and stress‐inducible expression.     

Molecular characterization of phytoplasma diseases of pepper in Turkey

During autumn, an extensive survey was conducted in pepper (Capsicum annum L.) in intensive cultivation areas of four provinces in southeastern Turkey (Adana, Kahramanmara?, Mersin and ?anl?urfa) in order to identify the causal agent (s) of phytoplasma‐like symptoms (chlorosis, little‐leaf, short internodes and stunting). DNA amplification by PCR and RFLP analysis using EcoRI restriction enzyme confirmed the presence of phytoplasmas in ?anl?urfa and Mersin, and consequently their possible association with the symptoms. Sequencing and phylogenetic analysis revealed that the isolate from ?anl?urfa had 99% sequence identity with “Candidatus Phytoplasma trifolii” (16SrVI) and is a member of the clover proliferation group (16SrVI‐A). Additionally, the isolate from Mersin had 96% sequence identity with “Candidatus Phytoplasma asteris” (16SrI). Importantly, gene sequence of the Mersin isolate shared <97.5% similarity to previously discovered “Ca. Phytoplasma” species. Consequently, the phytoplasma detected from Mersin could represent a new “Ca. Phytoplasma” species and to our knowledge, this is the first report of asteris‐like phytoplasmas infecting pepper in Turkey.     

Global pattern and controls of biological nitrogen fixation under nutrient enrichment: A meta‐analysis

Biological nitrogen (N) fixation (BNF), an important source of N in terrestrial ecosystems, plays a critical role in terrestrial nutrient cycling and net primary productivity. Currently, large uncertainty exists regarding how nutrient availability regulates terrestrial BNF and the drivers responsible for this process. We conducted a global meta‐analysis of terrestrial BNF in response to N, phosphorus (P), and micronutrient (Micro) addition across different biomes (i.e, tropical/subtropical forest, savanna, temperate forest, grassland, boreal forest, and tundra) and explored whether the BNF responses were affected by fertilization regimes (nutrient‐addition rates, duration, and total load) and environmental factors (mean annual temperature [MAT], mean annual precipitation [MAP], and N deposition). The results showed that N addition inhibited terrestrial BNF (by 19.0% (95% confidence interval [CI]: 17.7%?20.3%); hereafter), Micro addition stimulated terrestrial BNF (30.4% [25.7%?35.3%]), and P addition had an inconsistent effect on terrestrial BNF, i.e., inhibiting free‐living N fixation (7.5% [4.4%?10.6%]) and stimulating symbiotic N fixation (85.5% [25.8%?158.7%]). Furthermore, the response ratios (i.e., effect sizes) of BNF to nutrient addition were smaller in low‐latitude (<30°) biomes (8.5%?36.9%) than in mid‐/high‐latitude (≥30°) biomes (32.9%?61.3%), and the sensitivity (defined as the absolute value of response ratios) of BNF to nutrients in mid‐/high‐latitude biomes decreased with decreasing latitude (p ≤ 0.009; linear/logarithmic regression models). Fertilization regimes did not affect this phenomenon (p > 0.05), but environmental factors did affect it (p < 0.001) because MAT, MAP, and N deposition accounted for 5%?14%, 10%?32%, and 7%?18% of the variance in the BNF response ratios in cold (MAT < 15°C), low‐rainfall (MAP < 2,500 mm), and low‐N‐deposition (<7 kg ha^?1 year^?1) biomes, respectively. Overall, our meta‐analysis depicts a global pattern of nutrient impacts on terrestrial BNF and indicates that certain types of global change (i.e., warming, elevated precipitation and N deposition) may reduce the sensitivity of BNF in response to nutrient enrichment in mid‐/high‐latitude biomes.     

ICAM‐1 and MKL‐1 polymorphisms impose considerable impacts on coronary heart disease occurrence

This study was aimed to explore the correlation of intercellular adhesion molecule‐1 (ICAM‐1) K469E and megakaryoblastic leukaemia factor‐1 (MKL‐1) ?184C/T polymorphisms with the susceptibility to coronary heart disease (CHD) in the Chinese Han population. 100 CHD patients and 91 healthy people that had no blood connection with each other were enrolled in this case‐control study. ICAM‐1 and MKL‐1 polymorphisms were genotyped by polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) approach. Multiple logistic regression was used to analyse the correlation between polymorphisms of ICAM‐1 and MKL‐1 and CHD susceptibility. Differences of genotype and allele frequencies of the two SNPs between case and control groups were analysed by chi‐square test. Odds ratios (ORs) and 95% confidence intervals (CIs) were indicated relative susceptibility of CHD. The distributions of ICAM‐1 and MKL‐1 polymorphisms in each group conformed to Hardy‐Weinberg equilibrium (HWE). After adjusting for traditional risk factors, the TT genotype frequency of MKL‐1 ?184C/T polymorphism was found significantly higher in case group than in control group (P < .05). Meanwhile, T allele frequency increased in case group compared with control group, and the differences had statistical significance (P = .04, OR = 2.34, 95% CI = 1.34‐5.26). Logistic regression analysis in this study proved that smoking, hypertension, diabetes and triglyceride (TG) were all risk factors for CHD ICAM‐1 K469E polymorphism has no association with the onset of CHD. But MKL‐1 ?184C/T polymorphism is associated with the risk of CHD and T allele might be a susceptibility factor for CHD.     

Reversing expansion of Calamagrostis epigejos in a grassland biodiversity hotspot: Hemiparasitic Rhinanthus major does a better job than increased mowing intensity

Questions

Can hemiparasitic Rhinanthus major originating from a local population suppress the competitive clonal grass Calamagrostis epigejos and reverse its expansion in species‐rich semi‐natural grasslands? Does sowing seeds of R. major facilitate restoration of target meadow vegetation? Is R. major more beneficial for biodiversity restoration/conservation than increased mowing intensity, a conventional measure to suppress C. epigejos?

Location

?ertoryje National Nature Reserve, Bílé Karpaty (White Carpathians) Protected Landscape Area, Czech Republic.

Methods

We conducted a before‐after‐control‐impact experiment in meadow patches heavily infested by C. epigejos: eight blocks, each containing four plots with four treatment combinations: (1) traditional management, i.e. mowing once in summer, (2) mowing in summer and autumn (3) mowing in summer and seed sowing of R. major, (4) mowing in summer and autumn and seed sowing of R. major. Above‐ground biomass of C. epigejos and vegetation composition of each of the plots were monitored every year from 2013 to 2016. To assess the effects of treatments, we analysed biomass production of C. epigejos, herb layer cover and vegetation composition.

Results

Both sowing R. major and an additional autumn meadow cut significantly suppressed C. epigejos. Their effects were additive and of comparable size. Both treatments also had significant but markedly different effects on community composition. Rhinanthus major facilitated directional community composition change towards the regional Brachypodio‐Molinetum meadows. In contrast, increased mowing intensity significantly decreased frequency of threatened species, which however may have also been influenced by R. major.

Conclusions

Sowing of autochthonous R. major seeds was demonstrated as an efficient tool to suppress C. epigejos and facilitate community restoration. It can be combined with an additional meadow cut to further accelerate decline of the grass. The additional cut should however be used as a short‐term practice (1–2 years) only to minimize potential negative effects of its long‐term application on some threatened plant species. The effects of R. major are comparable to those of Rhinanthus alectorolophus reported previously. As a species occurring naturally in species‐rich dry grasslands, R. major has a broader and longer‐term application potential than R. alectorolophus in ecological restoration and conservation of these communities.     

Including albedo in time‐dependent LCA of bioenergy

Albedo change during feedstock production can substantially alter the life cycle climate impact of bioenergy. Life cycle assessment (LCA) studies have compared the effects of albedo and greenhouse gases (GHGs) based on global warming potential (GWP). However, using GWP leads to unequal weighting of climate forcers that act on different timescales. In this study, albedo was included in the time‐dependent LCA, which accounts for the timing of emissions and their impacts. We employed field‐measured albedo and life cycle emissions data along with time‐dependent models of radiative transfer, biogenic carbon fluxes and nitrous oxide emissions from soil. Climate impacts were expressed as global mean surface temperature change over time (?T) and as GWP. The bioenergy system analysed was heat and power production from short‐rotation willow grown on former fallow land in Sweden. We found a net cooling effect in terms of ?T per hectare (?3.8 × 10^–11 K in year 100) and GWP₁₀₀ per MJ fuel (?12.2 g CO₂e), as a result of soil carbon sequestration via high inputs of carbon from willow roots and litter. Albedo was higher under willow than fallow, contributing to the cooling effect and accounting for 34% of GWP₁₀₀, 36% of ?T in year 50 and 6% of ?T in year 100. Albedo dominated the short‐term temperature response (10–20 years) but became, in relative terms, less important over time, owing to accumulation of soil carbon under sustained production and the longer perturbation lifetime of GHGs. The timing of impacts was explicit with ?T, which improves the relevance of LCA results to climate targets. Our method can be used to quantify the first‐order radiative effect of albedo change on the global climate and relate it to the climate impact of GHG emissions in LCA of bioenergy, alternative energy sources or land uses.     

The decomposition of triphenylimidazole‐para‐acetate follows specific base catalysis and can be conveniently followed by fluorescence

The kinetics of the decomposition reaction of 4‐(4,5‐diphenyl‐1H‐imidazol‐2‐yl)phenyl acetate ( 1 ) in basic alcoholic media was investigated, using a simple fluorescence (FL) spectrophotometric procedure. The process was conveniently studied using FL, since the triphenylimidazole‐derived ester 1 and its reaction products (the corresponding phenol 2 and phenolate 2 ^? ) are all highly fluorescent (Φ_FL > 37%). By carefully selecting excitation and emission wavelengths, observed rate constants k₁ in the order of 10^?3 to 10^?2 s^?1 were obtained from either reactant consumption (λ_ex = 300 nm, λ_em = 400 nm) or product formation (λ_ex = 350 nm, λ_em = 475 nm); these were shown to be kinetically equivalent. Intensity‐decay time profiles also gave a residual FL intensity parameter, shown to be associated to the distribution of produced species 2 and 2 ^? , according to the basicity of the medium. Studying the reaction in both methanol (MeOH) and isopropanol (iPrOH), upon addition of HO^?, provided evidence that the solvent's conjugate base is the active nucleophilic species. When different bases were used (tBuO^?, HO^?, DBU and TEA), bimolecular rate constants k_bim ranging from 4.5 to 6.5 L mol^?1 s^?1 were obtained, which proved to be non‐dependent on the base pK_aH, suggesting specific base catalysis for the decomposition of 1 in alcoholic media.     

High‐efficient and precise base editing of C•G to T•A in the allotetraploid cotton (Gossypium hirsutum) genome using a modified CRISPR/Cas9 system

The base‐editing technique using CRISPR/nCas9 (Cas9 nickase) or dCas9 (deactivated Cas9) fused with cytidine deaminase is a powerful tool to create point mutations. In this study, a novel G. hirsutum‐Base Editor 3 (GhBE3) base‐editing system has been developed to create single‐base mutations in the allotetraploid genome of cotton (Gossypium hirsutum). A cytidine deaminase sequence (APOBEC) fused with nCas9 and uracil glycosylase inhibitor (UGI) was inserted into our CRISPR/Cas9 plasmid (pRGEB32‐GhU6.7). Three target sites were chosen for two target genes, GhCLA and GhPEBP, to test the efficiency and accuracy of GhBE3. The editing efficiency ranged from 26.67 to 57.78% at the three target sites. Targeted deep sequencing revealed that the C→T substitution efficiency within an ‘editing window’, approximately six‐nucleotide windows of ?17 to ?12 bp from the PAM sequence, was up to 18.63% of the total sequences. The 27 most likely off‐target sites predicted by CRISPR‐P and Cas‐OFFinder tools were analysed by targeted deep sequencing, and it was found that rare C→T substitutions (average < 0.1%) were detected in the editing windows of these sites. Furthermore, whole‐genome sequencing analyses on two GhCLA‐edited and one wild‐type plants with about 100× depth showed that no bona fide off‐target mutations were detectable from 1500 predicted potential off‐target sites across the genome. In addition, the edited bases were inherited to T1 progeny. These results demonstrate that GhBE3 has high specificity and accuracy for the generation of targeted point mutations in allotetraploid cotton.     

Metabolic engineering of Escherichia coli for agmatine production

Agmatine is a kind of important biogenic amine. The chemical synthesis route is not a desirable choice for industrial production of agmatine. To date, there are no reports on the fermentative production of agmatine by microorganism. In this study, the base Escherichia coli strain AUX4 (JM109 ?speC ?speF ?speB ?argR) capable of excreting agmatine into the culture medium was first constructed by sequential deletions of the speC and speF genes encoding the ornithine decarboxylase isoenzymes, the speB gene encoding agmatine ureohydrolase and the regulation gene argR responsible for the negative control of the arg regulon. The speA gene encoding arginine decarboxylase harboured by the pKK223‐3 plasmid was overexpressed in AUX4, resulting in the engineered strain AUX5. The batch and fed‐batch fermentations of the AUX5 strain were conducted in a 3‐L bioreactor, and the results showed that the AUX5 strain was able to produce 1.13 g agmatine L^?1 with the yield of 0.11 g agmatine g^?1 glucose in the batch fermentation and the fed‐batch fermentation of AUX5 allowed the production of 15.32 g agmatine L^?1 with the productivity of 0.48 g agmatine L^?1 h^?1, demonstrating the potential of E. coli as an industrial producer of agmatine.     

Isolation and characterization of eight microsatellite loci for the Neotropical freshwater catfish Pimelodella chagresi (Teleostei: Pimelodidae)

We report eight (CA)_10?35 unlinked microsatellite loci from the Neotropical freshwater catfish, Pimelodella chagresi (Siluriformes: Pimelodidae). These loci were characterized with 23 individuals collected in Panama. Number of alleles per locus varied from 7 to 23 (mean = 12.9) and observed heterozygosity ranged from 0.522 to 0.909 (mean = 0.732). These loci will be used to investigate the existence of cryptic species within the P. chagresi clade, and to study fine‐scale population structure.     

Preliminary evidence for a two‐for‐one deal: Wetland restoration for a threatened frog may benefit a threatened bat

Habitat restoration is an integral feature of wildlife conservation. However, funding and opportunities for habitat restoration are limited, and therefore, it is useful for targeted restoration to provide positive outcomes for non‐target species. Here, we investigate the possibility of habitat creation and management benefitting two threatened wetland specialists: the Green and Golden Bell Frog (Litoria aurea) and the Large‐footed Myotis (Myotis macropus). This study involved two components: (i) assessing co‐occurrence patterns of these species in a wetland complex created for the Green and Golden Bell Frog (n = 9) using counts, and (ii) comparing foraging activity of Large‐footed Myotis in wetlands with low and high aquatic vegetation (n = 6 and 7, respectively) using echolocation metres. Since Large‐footed Myotis possesses a unique foraging behaviour of trawling for aquatic prey, we hypothesised that foraging activity of this species would be higher in wetlands with low aquatic vegetation coverage. Additionally, we provide observations of its potential prey items. We identified one created wetland where both species were found in relatively high numbers, and this wetland had a permanent hydrology, was free of the introduced fish Gambusia (Gambusia holbrooki) and had low aquatic vegetation coverage. We also found that Myotis feeding activity was significantly higher in low aquatic vegetation coverage wetlands (x? = 65.72 ± 27.56 SE) compared to high (x? = 0.33 ± 0.33 SE, P = 0.0000). Although this is a preliminary study, it seems likely that Green and Golden Bell Frog and Large‐footed Myotis would gain mutual benefit from wetlands that are constructed to be permanent, that are Gambusia free, low in aquatic vegetation coverage, and are located in close to suitable roosting habitat for Large‐footed Myotis. We encourage adaptive aquatic vegetation removal for Green and Golden Bell frog as this may have benefits for Large‐footed Myotis. The evidence suggests that the former may be a suitable umbrella species for the latter.     

Winter hardiness of Miscanthus (II): Genetic mapping for overwintering ability and adaptation traits in three interconnected Miscanthus populations

Miscanthus × giganteus (M×g) is the primary species of Miscanthus for bioenergy feedstock production. The current leading biomass cultivar, M×g ‘1993‐1780’, is insufficiently adapted in temperate regions with cold winters such as USDA hardiness zone 5 (average annual minimum temperature of ?28.9 to ?23.3°C) or lower. Three interconnected Miscanthus F₁ populations that shared a common parent were planted in a replicated field trial at Urbana, IL (hardiness zone 5b; average annual minimum temperature of ?26.1 to ?23.3°C) in spring 2011. The winter of 2013–2014 was especially cold in Urbana, with a minimum soil temperature at 10 cm of ?6.2°C and a minimum air temperature of ?25.3°C, giving us an opportunity to evaluate hardiness on established year‐3 plants. The parent in common to all three populations, M. sinensis ssp. condensatus ‘Cosmopolitan’, is native to maritime southern Japan, and in Urbana, it is winter‐damaged most years. In contrast, the three other parents, M. sacchariflorus ‘Robustus’ (MapA), M. sinensis ‘Silberturm’ (MapB), and M. sinensis ‘November Sunset’ (MapC), are typically winter hardy in Urbana. Nearly all MapA progeny plants survived and grew vigorously in spring 2014, whereas in MapB and MapC, many progeny plants did not survive the winter, and most of the survivors were severely damaged, with poor vigor. Negative correlations between overwintering ability and spring regrowth date and autumn dormancy date suggested that the genotypes most likely to survive winters were those that emerged early in spring and/or went dormant early in autumn. Using joint‐population analysis, we identified 53 quantitative trait loci (QTLs) for nine adaptation traits, including nine QTLs for overwintering ability and 11 for spring hardiness scores. Many biologically intuitive candidate genes were observed within or near the QTLs detected in this study, suggesting their validity and potential for further study.     

Survivorship of geographic Pomacea canaliculata populations in responses to cold acclimation

1. Pomacea canaliculata, a freshwater snail from South America, has rapidly established natural populations from south to north subtropical region in China, since its original introductions in the 1980s. Low temperature in winter is a limiting factor in the geographic expansion and successfully establishment for apple snail populations. There have been some studies on population level of low temperature tolerance for P. canaliculata, yet little is quantified about its life‐history traits in responses to cold temperatures. Whether these responses vary with the acclimation location is also unclear. We investigated the survivorship and longevity of P. canaliculata in responses to cold temperatures and examine whether these responses vary with the location and snail size. We hypothesized that survival of the snails depends on their shell height and the level of low temperature, and P. canaliculata population from the mid-subtropical zone may exhibit the highest viability over the cold thermal range.
2. We sampled P. canaliculata populations from five latitude and longitude ranges of subtropical China: Guangzhou population in southernmost (SM‐GZ), three populations of Yingtan (MR‐YT), Ningbo (MR‐NB), Ya'an (MR‐YA) in midrange, and Huanggang population in northernmost (NM‐HG) subtropical zone. For each P. canaliculata population, survival and longevity at six cold acclimation temperature levels (12, 9, 6, 3, 0, and ?3°C) were quantified, and the effects of location and shell height were examined.
3. The MR‐YA population from mid-subtropical zone of China exhibited the highest survival rate and prolonged survival time regardless of the temperature acclimation treatments, whereas the SM‐GZ population from southern subtropical was the most sensitive to cold temperatures, particular temperatures below 9°C. No individuals of the SM‐GZ population could survive after stressed for 30 days (3°C), 5 days (0°C) and 2 days (?3°C), respectively. For each experimental P. canaliculata population held at 3, 0, and ?3°C, individuals with intermediate shell height of 15.0–25.0 mm had significantly higher survivals.
4. The results highlight a request of a more thorough investigation on acclimation responses in each of the life table demographic parameters for P. canaliculata, and pose the question of whether natural selection or some genetic changes may have facilitated adaptation in invasive locations.

     

The efficacy and safety of azithromycin in asthma: A systematic review

Azithromycin is a potential therapeutic choice for asthma control, which is a heterogeneous airway inflammatory disease. Because of variable findings, we intend to evaluate the therapeutic effect and safety of azithromycin in asthma. Databases, including PubMed, EMBASE, Cochrane, and CNKI until 31 December 2017, were searched to identify available randomised controlled trials regarding azithromycin treatment for asthma. We identified seven studies involving 1520 cases that met our criteria. The mean difference for lung function (FEV₁, FVC, PEF), symptom assessment (ACQ, AQLQ), airway inflammation, and risk ratios for adverse events were extracted. Chi‐square and I² tests were applied to evaluate the heterogeneity among the studies towards each index with a random effect model or a fixed effect model. Pooled analysis shows that azithromycin administration results in no significant improvement in FEV₁ (MD: 0.09, 95% CI ?0.10 to 0.29, P = 0.36), PEF (MD: 11.76; 95% CI, ?2.86 to 26.38, P = 0.11), total airway inflammatory cells (MD: ?0.29; 95% CI, ?1.38 to 0.80, P =  0.60), ACQ (MD: 0.05; 95% CI, ?0.08 to 0.19, P = 0.44), and AQLQ (MD: 0.12; 95% CI, ?0.02 to 0.26, P =  0.10). Moreover, no significant difference was detected in adverse events (Risk ratio 0.99; 95% CI, 0.82‐1.19, P = 0.90). These findings demonstrate no beneficial clinical outcome of azithromycin in asthma control, and we propose that further prospective cohorts are warranted.