Over thirty species of littoral marine Gammaridea occur along the coasts of the North Atlantic. From one to several species can coexist in a single region. There is an evident, inverse relationship between egg incubation time and temperature (from 14 to > 120 days) and consequent trends in the size of the animals on reaching maturity (from 5 mm in warmer waters to 30 mm in the coldest ones) and in lifespan (from < 6 months to > 5 years). Littoral gammarids are a good example of the shrinking size effect of increasing temperatures and size-related species diversity. In large species the annual cohorts of the population (3 to 5 annual size groups) functionally replace the adults of smaller species. The ongoing warming of the European Arctic seas may extend the distribution limits of boreal species so that more Gammarus species may appear on northern coasts hitherto occupied by just one or at most two species.

Domestic ducks are considered to have been tamed from the mallard or a descendant of the mallard and the spot-billed duck. Domestic ducks show remarkable phenotypic variation in morphology, physiology and behaviour. However, the molecular genetics of the origin and phenotypic variation of ducks are still poorly studied. Here, we present mallard and spot-billed genomes and perform whole-genome sequencing on eight domestic duck breeds and eight wild duck species. Surprisingly, analyses of these data support a model in which domestic ducks diverged from their closest wild lineage (mallard ducks and spot-billed ducks) at the last glacial period (LGP, 100-300 kilo years ago (Kyr)). The wild lineage further speciated into mallard ducks and spot-billed ducks approximately 70 Kyr, whereas the domestic lineage population decreased through the LGP. A scan of wild duck genomes compared with domestic duck genomes identified numerous loci that may have been affected by positive selection in ancestral wild ducks after their divergence from domestic lineages. Function analyses suggested that genes usually affecting organ development and energy metabolism may involve long-distance flight ability. Further selective sweep analyses identified two genes associated with egg production and three genes related to feeding modulation under selection in domestic ducks. These analyses unravel a distinct evolutionary pattern of ducks and two wild duck de novo genomes, thus providing a novel resource for speciation studies.

BLK belongs to the family of SRC kinases (SFKs), and are diagnosed with the aid of the presence of an SH3 and SH2 regulatory domains of N-terminal to the catalytic kinase domain. BLK in signalling has a vast position in transmitting alerts via immunoglobulins and ends in pro-B to pre-B conversion, and in signalling for boom arrest and apoptosis downstream of the B-cellular receptor. We have performed a series of computational analysis on various aspects on BLK viz phylogenetic analysis, domain analysis, secondary structure prediction, charge distribution, prediction of the antigenic region and have executed structural analysis by first structure modelling and then its refinement, and active site prediction for better understanding of the human BLK as a drug target. Our study includes a detailed analysis and graphical representation of different domains, charge distribution, prediction of the antigenic region etc with corresponding sequence and its secondary structure for the pharmacological aspect of BLK which observed that Tulipa suaveolens is the most outed clade in the BLK and Human BLK is found to be very closed to Pongo Abelii through the phylogenetic tree assessment.

Objective: To contribute to the validation of the self-administered ALSFRS-R - European Portuguese version (ALSFRS-R: EP), and to analyse its reliability in order to enable its use by ALS patients in Portugal. Methods: The study was developed in 3 phases. Phase 1: Translation and cultural adaptation of the self-administered ALFRS-R; Phase 2: Content validation by study participants; Phase 3: Analysis of internal consistency and test - retest reliability. Results: According to a panel of experts (N=6), a CVI of 100% were obtained for all ALSFRS-R: EP items. A sample of 18 people with ALS (13 male) fulfilled the test and evaluated its items regarding its clarity, comprehension, difficulty and relevance, obtaining values with an average between 8.6 and 8.9, 8.7 and 8.9, 8.5 and 8.8, and 8.5 and 8.9, respectively. The instrument total score and its subscales presented good internal consistency (Cronbach’s α: from 0.72 to 0.92 in the test; from 0.70 to 0.95 in the retest) and reliability (Kendall tau: from 0.575 to 0.990). Considering the scale total score for the test, no statistical differences were observed between females and males nor between bulbar ALS and medullar ALS. The correlation between the total score and age showed to be significant and negative (-0.53). Conclusions: The Self-Administered ALSFRS-R: EP version was successfully translated, validated, and presented good to excellent reliability results. It will allow its use by Portuguese ALS patients and enabling their health professionals to monitor the disease progression at home.

The article From Electrica to Invariant Automatica, with subtitle System and Invariant Electric Model, compares well-known power equations and other energy equations, regardless of the physical nature of the energy (mechanical, thermal, thermonuclear etc.) and thus the author creates a universal measure for the concept of energy. On this basis, the universal concept of a system is defined as a pair of two elements of energy source and energy consumer. The user is a production facility, at the output of which a product of certain qualities (mechanical dimensions, temperature, pressure, concentration, pH, etc.) appears. The control of the energy flow to it depends functionally on these qualities and, in addition, on the efficiency of the system (speed, flow rate, number of objects per unit of time, etc.). In the second part of the article with subtitle Electromechanical Dualism. The Universality of Energetic Equations, with the help of modern altebra, is proved the versatility of the electrical model of the system

A semiclassical phase-space perspective of band- and topological-insulator regimes of 2D Dirac materials, and normal- and superradiant-phases of atom-field interacting models is given in terms of delocalization, entropies, and quantum correlation measures. From this point of view, the low-energy limit of tight-binding models describing the electronic band structure of topological 2D Dirac materials like phosphorene and silicene with tunable band gaps, share similarities with Rabi-Dicke and Jaynes-Cummings atom-field interaction models, respectively. In particular, the edge state of 2D Dirac materials in the topological insulator phase exhibits a Schrödinger cat structure similar to the ground state of two-level atoms in a cavity interacting with a one-mode radiation field in the superradiant phase. Delocalization seems to be a common feature of topological insulator and superradiant phases.

This is a follow-on paper from the Hutchison group, expanding on some previous work looking at correlations of molecular energy from a variety of levels of theory with results from high-level ab initio calculations. A new addition in this paper is a small set of ML methods, a welcome addition to the forcefield and electronic structure methods usually used in comparisons of this kind.The paper presents some interesting results, but is riddled with missing or misattributed data, typos, grammatical errors (particularly agreements for single and plural nouns) and errors in the references. The paper should be carefully corrected before resubmission.The key omission in the paper is any attempt to provide confidence in the deductions made about the differences in accuracy between the methods compared. Confidence intervals on each of the estimators, estimates of success rates and their errors, and pairwise hypothesis tests, at a minimum, must be added before publication. With this data in hand the new version can make quantitative estimates of the differences between the methods.

Despite ca. seven decades of theoretical elaboration since Peter Medawar’s foundational ‘An Unsolved Problem of Biology’, we argue that the fundamental problem of the evolution of senescence, i.e. the increasing risk of mortality and decline in reproduction with age after maturity, remains unsolved. Theories of senescence predict the inescapability of senescence, or its universality at least among species with a clear germ-soma barrier. Here, using demographic information for 475 multicellular species, we exemplify the discrepancy between these theoretical predictions and currently available data. We derive age-based trajectories of mortality and reproduction whose form cannot be satisfactorily explained by the theories of senescence, and show that species’ may often display senescence for one fitness component but not the other. We propose that theories of senescence must be extended beyond merely individual chronological age; size, the species’ ecological context, and kin selection may all play hidden, yet integral roles in shaping patterns of senescence.

Objective: The subsequent reproductive events induced by early age at menarche (AAM) are tightly linked to immune dysfunction. This study aimed to analyze whether immune functions mediate the association between AAM and subsequent reproductive performance. Design: a follow-up and Mendelian Randomization (MR) study. Setting: A women’s and Children’s hospital in Shenzhen, China. Population: Sixty-eight healthy reproductive Chinese women were admitted to pre-pregnancy physical examinations. Methods: Pre-pregnancy immune functions were analyzed by flow cytometry. Subsequent reproductive performance was studied by a 15-month follow up. The associations of immune functions with AAM or pregnancy status were analyzed. Lastly, the important association was further validated by a two-sample MR test using public data. Main Outcome Measures: Miscarriages, thyroid function at early pregnancy, and metabolic indexes at mid pregnancy. Results: We found that AAM was negatively associated with Tfh1/Tfh2 ratio (Spearman r=-0.283, P=0.019). Moreover, this pre-pregnancy index was positively associated with TSH at early pregnancy (Spearman r=0.363, P=0.032), a risk for spontaneous miscarriage (adjusted Relative risk (RR)=12.25, 95% confidence interval (CI)=1.72-87.46, P=0.013), and a shorter time to miscarriage (42 days vs. 115 days, log-rank P=0.038). Moreover, the MR test showed that 84 AAM-related SNPs can explain 19% of variance in PD1- naïve Tfh cells (Directionality P=4.28×10-10); LIN28B and chromosome 9p12 (LINC01505, TAL2 and TMEM38B) were their share genetic factors. Conclusion: The present study implied that Tfh cells might mediate the process of early AAM-induced reproductive events. Larger population studies and functional studies are warranted. Funding: None.

MYB transcription factors are important in abiotic stress responses; however, the detailed mechanisms are unclear. Tamarix hispida contains multiple MYB genes. The present study characterized T. hispida MYB (ThMYB8) during salt stress using transgenic T. hispida and Arabidopsis assays. ThMYB8 overexpression and ThMYB8 RNAi analysis demonstrated that ThMYB8 enhanced the salt stress tolerance. Transgenic Arabidopsis overexpressing ThMYB8 displayed significantly increased root growth, fresh weight, and seed germination rate compared with that of the wild-type. Physiological parameters analysis in T. hispida and Arabidopsis showed that ThMYB8 overexpressing plants had the lowest levels of O2-, H2O2, cell death, malondialdehyde, and electrolyte leakage. Overexpression of ThMYB8 regulated Na+ and K+ concentrations in plant tissues while maintaining K+/Na+ homeostasis. Analysis using qRT-PCR and ChIP-PCR identified possible downstream ThMYB8-regulated genes. ThMYB8 regulated the expression of ThCYP450-2 (cytochrome p450-2), Thltk (leucine-rich repeat transmembrane protein kinase), and ThTIP (aquaporin TIP) by binding to the MBSI motif (‘CAACTG’) in their promoters. The results indicated that ThMYB8 enhanced salt stress tolerance in T. hispida by regulating gene expression related to the activation of stress-associated physiological changes, such as enhanced reactive oxygen species scavenging capability, maintaining K+/Na+ homeostasis, and decreasing the malondialdehyde content and lipid peroxidation cell membranes.

Objective: Clinical trials provide fundamental evidence used to inform healthcare decisions at patient- and population levels. It is therefore important that trials evaluate outcomes considered relevant by patients and relevant stakeholders. Although validated tools assessing other aspects of trial integrity exist, there is no tool for assessing the breadth and completeness of outcomes being measured. We have developed the Comprehensiveness of Outcome Reporting (COR) Tool to assist systematic reviewers and trialists in evaluating and choosing trial outcomes within the dynamic context of obstetrics. Methods: We identified five core outcome areas – mortality, clinical/physiological, functioning/life-impact, resource-use, and adverse events – from a published taxonomy for outcomes in medical research, and programmed an excel-based tool capable of producing a heatmap to enable users to visualize whether trial outcomes appropriately represent all outcomes areas for both mother and fetus. We used a mock-heatmap to demonstrate the tool’s utility in assessing comprehensiveness of outcome reporting in obstetric trials. Results: This excel-based tool guides users through a series of simple questions regarding the clinical trial(s) being assessed, producing a heatmap output that depicts the spread of reported outcome areas. Trends are readily interpreted with a heatmap, with over/under-reported maternal and fetal-neonatal outcome areas clearly highlighted. Conclusions: Comprehensive reporting of outcomes is necessary to ensure that interventions truly result in improved outcomes in all core areas. The COR Tool will enable systematic reviewers and trialists to determine and select outcomes with more breadth and completeness, encouraging transparency and the drawing of valid clinical conclusions. Funding: None

Abstract Objective To evaluate the clinical diagnostic validity of carbon nanoparticle suspension (CNS) in sentinel lymph node biopsy (SLNB) for assessing lymphatic spread of early-stage cervical cancer. Design A prospective study. Setting and population 356 cases. Methods We enrolled 356 stage Ia2-IIa2 cervical cancer patients to undergo SLNB using CNS followed by systematic pelvic lymphadenectomy. All lymph node specimens were assessed using conventional histopathologic ± pathologic ultrastaging analyses. Main outcome measures SLN detection rate (DR), clinical diagnostic validity, and various related factors were analysed. Results CNS identified 1456 SLNs in 325 patients. The overall SLN DR was 91.29%. A significantly higher DR was found for patients with tumours <20 mm (97.75% vs. 71.91%; p = 0.000). Two patients had false-negative results, accounting for 0.615% of patients who had successful SLN detection. SLNB with CNS had sensitivity of 92.86%, false-negative rate (FNR) of 7.14%, and negative predictive value (NPV) of 99.29%. Importantly, sensitivity (100%), NPV (100%), and FNR (0%) were improved when testing the subgroup of patients with tumours <20 mm (267 cases). There were no observed differences in DR based on pathologic type or grade, stage, depth of stromal invasion, surgical approach, menopausal status, or prior treatment with chemotherapy (p > 0.05). Conclusions SLNB with CNS results in favourable DR, sensitivity, and NPV for women with early-stage cervical cancer with small tumour sizes. SLNB with CNS is safe, feasible, and relatively effective for guiding precise surgical treatment of early-stage cervical cancer. Keywords SLNB, CNS, early-stage cervical cancer

Accurate and testable species delimitation hypotheses are essential for measuring, surveying and managing biodiversity. Today, taxonomists often rely on mitochondrial DNA barcoding to complement morphological species delimitations. Although COI barcoding has largely proven successful in assisting identifications for most animal taxa, there are nevertheless numerous cases where mitochondrial barcodes do not necessarily reflect the species history. For instance, what is regarded as one single species can be associated with two distinct DNA barcodes, which can point either to cryptic diversity or to deep within-species mitochondrial divergences with no reproductive isolation. In contrast, two or more species can share barcodes, for instance due to mitochondrial introgression. These intrinsic limitations of mitochondrial DNA barcoding can only be addressed with nuclear genomic markers, which are expensive, labour intensive, poorly repeatable, and often require high-quality DNA. To overcome these limitations, we examined the use of ultraconserved nuclear genetic elements (UCEs) as a quick and robust genomic approach to address such problematic cases of species delimitation. This genomic method was assessed using six different bee species complexes suspected to harbour cryptic diversity, mitochondrial introgression, or mitochondrial paraphyly. The sequencing of UCEs recovered between 686 and 1860 homologous nuclear loci and provided explicit species delimitation hypotheses in all investigated species complexes. These results provide strong evidence for the suitability of UCEs as a fast method for species delimitation even in recently diverged lineages. Furthermore, this study provided the first conclusive evidence for both mitochondrial introgression among distinct species, and mitochondrial paraphyly within a single bee species.

Objective: High data quality is essential to ensure the validity of clinical and research inferences based on it. However, these data quality assessments are often missing even though these data are used in daily practice and research. Our objective was to evaluate the data quality of our high-resolution electronic database (HRDB) implemented in our pediatric intensive care unit (PICU). Design: A prospective validation study of a HRDB. Setting: A 32-bed pediatric medical, surgical and cardiac PICU in a tertiary care freestanding maternal-child health center in Canada. Population: All patients admitted to the PICU with at least one vital sign monitored using a cardiorespiratory monitor connected to the central monitoring station. Interventions: None Measurements and Main Results: Between June 2017 and August 2018, data from 295 patient days were recorded from medical devices and 4,645 data points were video recorded and compared to the corresponding data collected in the HRDB. Statistical analysis showed an excellent overall correlation (R2=1), accuracy (100%), agreement (bias=0, limits of agreement=0), completeness (2% missing data) and reliability (ICC=1) between recorded and collected data within clinically significant pre-defined limits of agreement. Divergent points could all be explained. Conclusions: This prospective validation of a representative sample showed an excellent overall data quality.

Maternal effects have been well documented for offspring morphology and life history traits in plants and terrestrial animals, yet little is known about maternal effects in corals. Further, few studies have explored maternal effects in gene expression. In a previous study, F1 interspecific hybrid and purebred larvae of the coral species Acropora tenuis and A. loripes were settled and exposed to ambient or elevated temperature and pCO2 conditions for seven months. At this stage, the hybrid coral recruits from both ocean conditions exhibited strong maternal effects in several fitness traits. We conducted RNA-sequencing on samples from the same experiment and showed that gene expression of the hybrid Acropora also showed clear maternal effects. Only 40 genes were differentially expressed between hybrids and their maternal progenitor. In contrast, ~2000 differentially expressed genes were observed between hybrids and their paternal progenitors, and between the reciprocal F1 hybrids. These results indicate that maternal effects in coral gene expression can be long-lasting. Unlike findings from most short-term stress experiments in corals, no genes were differentially expressed in the hybrid nor purebred offspring after seven months of exposure to elevated temperature and pCO2 conditions.

Phaeodactylum tricornutum is a lipid-rich marine diatom that contains a high level of omega-3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA). In an effort to reduce costs for large-scale cultivation of this microalga, we first established a modified BBM medium (0.3 x strength BBM with 90% reduced phosphate content) to replace the traditional F/2 medium. P. tricornutum could grow in extremely low phosphate concentrations (25 µM), without compromising the EPA content. In the presence of sea salts, silicate addition was not necessary for high rate growth, high EPA content or lipid accumulation in this species. Using urea as the sole nitrogen source tended to increase EPA contents (by 24.7%) while not affecting growth performance. The use of sea salts, rather than just sodium chloride led to significantly improved biomass yields (20% increase) and EPA contents (46-52% increase), most likely because it supplied sufficient essential elements such as magnesium. A salinity level of 35 ppt led to significantly higher biomass yields compared to 20 ppt, but salinity had no significant influence on EPA content that reached high average levels of 51.8% of total fatty acids during exponential growth phase at 20 ppt in modified BBM medium with sea salts.

The prediction of future land cover changes is an important step in proper planning and management of watersheds. Various methods exist for this purpose. In this study, land cover changes were investigated in the Hable-Rud River basin in Iran, an arid and semi-arid region, using remote sensing and Geographic Information Systems (GIS). First, a supervised classification technique was applied to Landsat images acquired for 1986, 2000 and 2017 using the maximum likelihood method. Then, using pixel-by-pixel change detection, the land cover changes were predicted for 2017 and 2040 using a Cellular Automata (CA)-Markov model. The descriptive variables used included slope, aspect, elevation, and calculated distances from various land features such as rivers, roads, industrial areas, residential areas, saline land, and land in agricultural production. The predictions for 2017 were validated using the derived map from a Landsat image of 2017 with a resulting standard Kappa index of 0.74. According to the prediction results for 2040, the areas of rangeland and saline land will increase by approximately 6.5% and 2%, respectively, whereas the areas of bare land and agricultural land will decrease by approximately 6% and 2%, respectively. Moreover, the analysis of historical records since 1986 showed that the annual streamflow and precipitation have reduced by almost 44% and 29%, respectively. The reductions, particularly to streamflow, can be attributed largely to agriculture expansion, rapid population growth, and industrial developments. The analysis of the results indicates a need for more effective design, planning, and development of land cover policies for ecosystem protection.

Changes in the catchment scale water balance have important social implications for usable water now and in the future. Stream discharge is also directly related to radionuclides flux in the river water system. The aim of this study was to clarify the water balance in the Chernobyl Exclusion Zone (CEZ) under current and future climate conditions. A catchment scale hydrological model was used with long-term discharge data to project the future trend of radionuclides wash-off from the contaminated catchment at the CEZ in Ukraine. The Sakhan river catchment at the CEZ (51.41°N, 30.00°E) in Ukraine is one of the Pripyat river systems, and has a total surface area of 186.9 km2. We found that under the current climate, 84% of annual input (sum of rainfall and snowmelt) was consumed as evapotranspiration, and discharge was estimated to be 16%. In future climates, annual precipitation is expected to increase. However, a projected increase in the vapor pressure deficit led the consumption of precipitation as evapotranspiration and no significant increase in discharge. The study found that warmer winter and spring temperatures will decrease the snowfall, and increase the rainfall, but it was not enough to increase evapotranspiration. As a result, the peak of discharge shifted from April to March. The increase of future average discharge during the winter and spring came from a combination of (1) increasing rainfall in the winter and spring, and (2) relatively small levels of evapotranspiration, which enhanced the catchment scale water recharge in soil moisture and gave rise to greater discharge during winter and spring. The reduction of extreme river discharge from the hydrological projections could reduce the probability of high radionuclides concentration in the river water system in the future, owing to the reduction of surface runoff water from the contaminated surface soil and/or top layer of floodplain soils in the CEZ.

Bosnia and Herzegovina (BIH) is developing country facing extensive land degradation. As a post-conflict society with underlying poor socio-economic conditions, the value of land and its degradation status are not perceived to be an important problem neither a priority to address. BIH currently exists as a decentralized state, where land and land resources are under exclusive jurisdiction of two entities and one district, rather than state-level legislation. Complex land related administration between entities functioning independently from each other, and thus not sinchronized, leads to frequent political conflicts about land property, and limited data exchange which may exacerbate current land degradation. This paper investigates (i) the institutional setting and policy framework related to land, discussing the effectiveness and limitations of the current policies in this post-conflict society; (ii) stakeholders’ perception of land degradation under such complex institutional and policy structures; and (iii) the current state of land degradation, with a focus on soil erosion as one of the most important indicators in BIH. Communication and cooperation are major challenges for sustainable land management in post conflict societies. The existence of a policy framework is important, but not sufficient if implementation is weak and the perception of decision makers differs. Limited data on the impact of 1992 War on soil and land status represents an additional challenge to combat land degradation. Reliable national data on land degradation are crucial for assessment and development of strategic and policy frameworks devoted to those issues and to raise awareness on how to foster their implementation.

A document by Eugeniusz Gil. Click on the document to view its contents.

The temporal and spatial distribution of water resources over China has changed and may continue changing in the future under ongoing global warming. Scientific water resources management requires reliable forecasting of the change. Meanwhile, the performance of deep learning in achieving it has not been comprehensively explored. To fill this gap, deep learning, i.e., multilayer perceptron (MLP) in this study, is used to study the change of streamflow over China under climate changes. MLP is compared with other machine learning methods for investigating its strengths, and three river basins (i.e., Xiangxi, Jinghe and Zhongzhou) in central, northwestern and southeastern China, respectively are selected to represent hydrologic regimes over China. Four regional climate models are used to drive MLP for forecasting streamflow from 2021 to 2050 under two greenhouse-gas emission scenarios (i.e., RCPs 4.5 and 8.5). Modeling results show that MLP is more accurate than the other methods, especially in terms of peak streamflow volumes. Annual average temperature in the three basins will increase, while precipitation shows different changing trends. The simulation accuracies among the regional climate models (RCMs) are slightly different. Correspondingly, streamflow will increase, and the increments decrease from Jinghe, through Xiangxi, to Zhongzhou River Basins. Due to climate changes, flooding will become more frequent in Jinghe and Xiangxi River Basins, Jinghe River Basin will experience no runoff in winter, and the timing of peak runoffs in Zhongzhou River Basin will move forward. Compared with the RCP 4.5 scenario, the above trends are more obvious under the RCP 8.5 scenario.

The Arctic is warming at an unprecedented rate. One relatively under researched process is how seasonally frozen soils and changes thereof affect the water cycle. As frozen soils thaw, flow pathways within a watershed open, allowing for enhanced hydrologic connectivity between groundwater and rivers. As the connectivity of flow paths increase, the storage-discharge relationship of a watershed changes. The objective of this study is to quantify trends and spatio-temporal differences in the degree of linearity in the storage-discharge relationships for sixteen watersheds within Northern Sweden throughout the years of 1950 and 2018. We demonstrate a clear increase in non-linearity of the storage-discharge relationship over time for all catchments with twelve out of sixteen watersheds (75%) having a statistically significant increase in non-linearity. Springs have significantly more linear storage-discharge relationships than summer for twelve watersheds (75%), which supports the idea that seasonally frozen soil with a low degree of hydrological connectivity have a linear storage-discharge relationship. For the period considered, spring showed the greater change in storage-discharge relationship trends than summer, signifying changes in recessions are occurring during the thawing period. Separate storage-discharge analyses combined with preceding winter conditions demonstrated that especially cold winters with little snow yield springs and summers with more linear storage-discharge relationships. We show that streamflow recession analysis shows ongoing hydrological change of an arctic landscape as well as offers new metrics for tracking the change across arctic and sub-arctic landscapes.

Mulberry intercropped with alfalfa is a popular agroforestry system in China, which can produce high forage yields with high protein. To investigate the advantages of intercropping as well as the responses of mulberry-alfalfa intercropping systems to nitrogen application, we studied changes in the soil microbial communities and physiochemical properties in the rhizosphere of intercropped mulberry and alfalfa under nitrogen application. Nitrogen application increased available nitrogen contents (AN) and activities of urease (SUR) in rhizosphere soil of mulberry and alfalfa, but reduced soil organic canmatter (OM), irrespective of the cropping system (monoculture or intercropping). Nitrogen application increased soil pH in the rhizosphere of mulberry and alfalfa in monoculture, but reduced soil pH in the intercropped system. Nitrogen application and intercropping enhanced the soil water content (SWC) in mulberry rhizosphere soil and decreased SWC in alfalfa rhizosphere soil. We observed significant differences in the microbial communities inhabiting mulberry and alfalfa rhizosphere soils, indicating that mulberry and alfalfa are strategically complementary in terms of carbon sources. The Shannon-Weaver (H’), Simpson index (D), and McIntosh diversity (U) values were higher in rhizosphere soil of intercropped mulberry compared with mulberry in monoculture without nitrogen supply. There were no significant differences between mulberry in monoculture with nitrogen application (MNE) and mulberry intercropped alfalfa without nitrogen (M0). The results of the principal components analysis showed that MNE and M0 clustered in the scattered plots. We found no significant differences for H’ and D between monoculture and intercropping alfalfa systems, irrespective of the nitrogen application. Nitrogen application, intercropping, and intercropping with nitrogen increased the numbers of carbon sources, and relative use rates exceeded 4%. Nitrogen application and intercropping reduced the numbers of available carbon sources in alfalfa. Redundancy discriminatory analysis results suggest that pH and SWC were positively related with mulberry treatments and negatively with the alfalfa treatments.

The complex and enormous diversity of microbiome associated with plant roots is important for plant health and is shaped by numerous factors. This study aimed to unravel the effects of a pesticide-fertilizer combination on the rhizosphere microbiome of field-grown sugarcane. A field trial on sugarcane was conducted in Zhanjian city, Guangdong Province of China, and bulk soil and rhizosphere samples were collected 3 weeks after planting. We examined the effects of clothianidin and/or organic fertilizer treatments on the composition, diversity, and function of rhizosphere microbial communities using 16S rRNA gene and ITS1 gene amplicon sequencing. Compared with the controls (no pesticide or fertilizer used), the microbial community that resulted from treating with the pesticide-fertilizer (SPF) had a higher relative bacterial diversity and abundance, as well as contributing more comprehensive functions to sugarcane. The bacterial and fungal compositions at different taxonomic levels affected by clothianidin in the SPF and SP (with pesticide addition) were different from the effects experienced in the other treatments. With organic fertilizers added to SPF, the abundance of soil beneficial bacteria Bacillus, Paenibacillus, and Brevibacillus were highly improved, as well as the microbial function. It indicated that the SPF treatment could counteract the effects caused by mixing with clothianidin when compared to the SP treatment with only pesticide addition. Moreover, four bacterial genera including Dyella, Sphingomonas, Catenulispora, Mucilaginibacter, and Tumebacillus were significantly increased in the SPF and SP groups, which was reported to degrade clothianidin and could improve the soil health. The findings of the study provide insights into the interaction between the rhizosphere soil microbiome and a pesticide-fertilizer integration that may help improve application for pesticide-fertilizer in sugarcane fields.