Invasive alien ants(IAAs)are among the most aggressive,competitive,and widespread invasive alien species(IAS)worldwide.Wasmannia auropunctata,the greatest IAAs threat in the Pacific region and listed in“100 of the wo...Invasive alien ants(IAAs)are among the most aggressive,competitive,and widespread invasive alien species(IAS)worldwide.Wasmannia auropunctata,the greatest IAAs threat in the Pacific region and listed in“100 of the world’s worst IAS”,has established itself in many countries and on islands worldwide.Wild populations of W.auropunctata were recently reported in southeastern China,representing a tremendous potential threat to China’s agricultural,economic,environmental,public health,and social well-being.Estimating the potential geographical distribution(PGD)of W.auropunctata in China can illustrate areas that may potentially face invasion risk.Therefore,based on the global distribution records of W.auropunctata and bioclimatic variables,we predicted the geographical distribution pattern of W.auropunctata in China under the effects of climate change using an ensemble model(EM).Our findings showed that artificial neural network(ANN),flexible discriminant analysis(FDA),gradient boosting model(GBM),Random Forest(RF)were more accurate than categorical regression tree analysis(CTA),generalized linear model(GLM),maximum entropy model(MaxEnt)and surface distance envelope(SRE).The mean TSS values of ANN,FDA,GBM,and RF were 0.820,0.810,0.843,and 0.857,respectively,and the mean AUC values were 0.946,0.954,0.968,and 0.979,respectively.The mean TSS and AUC values of EM were 0.882 and 0.972,respectively,indicating that the prediction results with EM were more reliable than those with the single model.The PGD of W.auropunctata in China is mainly located in southern China under current and future climate change.Under climate change,the PGD of W.auropunctata in China will expand to higher-latitude areas.The annual temperature range(bio7)and mean temperature of the warmest quarter(bio10)were the most significant variables affecting the PGD of W.auropunctata in China.The PGD of W.auropunctata in China was mainly attributed to temperature variables,such as the annual temperature range(bio7)and the mean temperature of the warmest quarter(bio10).The populations of W.auropunctata in southern China have broad potential invasion areas.Developing strategies for the early warning,monitoring,prevention,and control of W.auropunctata in southern China requires more attention.展开更多
Agrometeorological disasters severely impact agriculture in Heilongjiang Province.Flood is one of the main agrometeorological disasters in Heilongjiang Province.The temporal change in flood events in Heilongjiang Prov...Agrometeorological disasters severely impact agriculture in Heilongjiang Province.Flood is one of the main agrometeorological disasters in Heilongjiang Province.The temporal change in flood events in Heilongjiang Province from 1986to 2015 was studied using Mann-Kendall and Morlet wavelet methods,respectively.The results of Mann-Kendall analysis showed that the disaster rates of flood gradually stabilized from 1986 to 2015 with a confidence level of 99%.The Morlet wavelet variance analysis revealed that disaster rates of flood changed periodically at time scales of 3a,7a and 18a in Heilongjiang Province during1986-2015.The dominant period of the variation of flood disaster rate was about 18a over the past 30 years.The flood disaster rates were indicated in a positive phase during the period of 2016-2020 by the fitting curve of Morlet wavelet analysis.The annual average flood disaster indexes of single station,during 1986-2015 years were calculated,according to the precipitation data at 31 stations in Heilongjiang Province and the GIS software was used to analyze the spatial change in flood disasters in Heilongjiang Province from1986-2015.The results demonstrated that the southwest area of Heilongjiang Province was highly hazardous region of flood.The flood indices in the northern part of Songnen Plain and southwest of Heilongjiang Province presented the increment trends.展开更多
Agricultural non-point source pollution is increasingly an important issue affecting surface water quality.Currently,the majority of the studies on nitrogen loss have focused on the agricultural field scale,however,th...Agricultural non-point source pollution is increasingly an important issue affecting surface water quality.Currently,the majority of the studies on nitrogen loss have focused on the agricultural field scale,however,the response of surface water quality at the watershed scale into the nitrogen loss at the field scale is poorly understood.The present study systematically reviewed the critical processes and major factors that nitrogen transport from farm fields to surface water bodies.The critical processes of farmland nitrogen entering surface water bodies involve the processes of nitrogen transport from farmland to ditches and the transformation processes of nitrogen during migration in ditches/rivers.Nitrogen transport from farmland to ditches is one of the prerequisites and critical processes for farmland nitrogen transport to surface water bodies.The transformation of nitrogen forms in ditches/rivers is an intermediate process in the migration of nitrogen from farmland to surface water bodies.Nitrogen loss from farmland is related to soil storage and exogenous inputs.Therefore,nitrogen input management should not only consider the current input,but also the contribution of soil storage due to the historical surpluses.Ditches/rivers have a strong retention capacity for nitrogen,which will significantly affect the process of farmland nitrogen entering surface water bodies.The factors affecting nitrogen transformation in river/ditches can be placed in four categories:(1)factors affecting hydraulic retention time,(2)factors affecting contact area,(3)factors affecting biological activity,and(4)forms and amount of nitrogen loading to river/ditches.Ditch systems are more biologically(including plants and microbes)active than rivers with biological factors having a greater influence on nitrogen transformation.When developing pollution prevention and control strategies,ecological ditches can be constructed to increase biological activity and reduce the amount of surplus nitrogen entering the water body.The present research should be valuable for the evaluation of environment impacts of nitrogen loss and the non-point source pollution control.展开更多
Cadmium(Cd)contamination in paddy soils means that the rice produced there may be unsafe for human consumption.A hydroponic study was conducted to enrich rice seedlings with zinc(Zn)or manganese(Mn),and the uptake and...Cadmium(Cd)contamination in paddy soils means that the rice produced there may be unsafe for human consumption.A hydroponic study was conducted to enrich rice seedlings with zinc(Zn)or manganese(Mn),and the uptake and transport characteristics of Cd in these Zn-and Mn-rich seedlings were subsequently investigated using a greenhouse pot trial.The results showed that hydroponic cultivation in 10-50μmol L^(-1) Zn(ZnSO_(4)·7 H_(2)O)or50-250μmol L^(-1) Mn(MnSO_(4)·H_(2)O)for 30 d had no significant impact on rice growth,while the accumulation of Zn and Mn was 7.31-18.5 and 25.4-47.7 times higher,respectively,than in the control(no Zn or Mn addition).The accumulation of Cd in the Zn-and Mn-rich rice plants was 26.3%-38.6% and 34.4%-44.5% lower than that in the control,respectively,and the translocation factors of Cd from roots to shoots also decreased by 23.3%-41.3% and 25.3%-37.0%,respectively,after transplanting to Cd-contaminated soils.Furthermore,the relative expression levels of OsIRT1(Oryza sativa iron-regulated transporter 1)were downregulated by 40.1%-59.3% and 16.0%-25.9%,respectively,in the Zn-and Mn-rich seedling roots.This downregulation may indicate a possible mechanism contributing to the reductions in Cd absorption.Field experiments confirmed that the Zn-and Mn-rich seedlings produced brown rice(unpolished rice grains)with significantly decreased concentrations of Cd(34.2%-44.4%).This study provides an innovative method for reducing the food safety risks from rice grown on slightly to moderately Cd-contaminated paddy soils.展开更多
The management of livestock waste is an effective way to achieve emission reduction and carbon fixation in agriculture and rural areas.At present,aerobic composting and anaerobic fermentation are widely used in livest...The management of livestock waste is an effective way to achieve emission reduction and carbon fixation in agriculture and rural areas.At present,aerobic composting and anaerobic fermentation are widely used in livestock waste treatment technology.In this study,pig manure management was taken as an example,a comprehensive environmental load index was constructed to quantitatively evaluate the environmental impacts of global warming,environmental acidification,eutrophication,and photochemical ozone synthesis during aerobic composting and anaerobic fermentation based on the life cycle assessment.The results showed that the potential values of aerobic composting and anaerobic fermentation were similar,and the order was global warming,environmental acidification,eutrophication,and photochemical ozone synthesis.Anaerobic fermentation contributed more to global warming,while aerobic composting contributed more to environmental acidification,eutrophication,and photochemical ozone synthesis.In addition,the environmental load index of aerobic composting was significantly higher than that of anaerobic fermentation.There were certainly regional differences in the environmental load index,and the environmental impact effect of anaerobic fermentation was low and more environmentally friendly.These findings provided a technical basis for livestock manure management in different regions of China,which was conducive to promoting animal husbandry emission reduction and carbon sequestration.展开更多
Semiochemical-based management strategies are important for controlling bark beetles,such as invasive Red Turpentine Beetle(Denroctonus valens),the causal agent for mass mortality of pine trees(Pinus spp.)in China.It ...Semiochemical-based management strategies are important for controlling bark beetles,such as invasive Red Turpentine Beetle(Denroctonus valens),the causal agent for mass mortality of pine trees(Pinus spp.)in China.It has been previously shown that the pheromone verbenone regulates the attack density of this beetle in a dose-dependent manner and that the gut bacteria of D.valens are involved in verbenone production.However,molecular functional verification of the role of gut bacteria in the pheromone production of D.valens is still lacking.To better understand the molecular function of gut bacterial verbenone production,we chose a facultative anaerobic gut bacterium(Enterobacter xiangfangensis)of D.valens based on its strong ability to convert cis-verbenol to verbenone,as shown in our previous study,and investigated its transcriptomics in the presence or absence of cis-verbenol under anaerobic conditions(simulating the anoxic environment in the beetle's gut).Based on this transcriptome analysis,aldehyde dehydrogenase(ALDH1)was identified as a putative key gene responsible for verbenone production and was knocked-down by homologous recombination to obtain a mutant E.xiangfangensis strain.Our results show that these mutants had significantly decreased the ability to convert the monoterpene precursor to verbenone compared with the wild-type bacteria,indicating that ALDH1 is primarily responsible for verbenone conversion for this bacterium species.These findings provide further mechanistic evidence of bacterially mediated pheromone production by D.valens,add new perspective for functional studies of gut bacteria in general,and may aid the development of new gene silencing-based pest management strategies.展开更多
In order to study the distribution of soil microplastics in the plastic film mulched farmland,the fluidization-centrifugation secondary density flotation method was optimized and improved to obtain the samples.The mai...In order to study the distribution of soil microplastics in the plastic film mulched farmland,the fluidization-centrifugation secondary density flotation method was optimized and improved to obtain the samples.The main components of microplastics were analyzed by Fourier spectrometer,and the surface morphology,porosity,particle size and abundance distribution characteristics of microplastics were studied by electron microscopy.The results showed that plastic mulch is the main source of microplastics.Its morphology mainly consists of fragments,fibers and particles.The size of debris microplastics was larger,with an average of 1.6300 mm,mainly distributed in the 0-300 mm cultivation layer while the size of particle micro plastic was smaller,with an average of 0.1400 mm.The width of fiber microplastic was 5-20μm,but the length could reach 0.2000-2.0000 mm,with an average particle size of 0.9200 mm.Fiber and granular microplastics could be seen in each soil layer.The surface layer of microplastics has a large number of pores characterized with a length of 50μm and a width of about 5-10μm,which might be an easy oxidation site for microplastics.This oxidation is continuous,making microplastics continuously decompose into smaller particles.The abundance of microplastics is negatively correlated with soil depth.The average abundance of microplastics is 161.50±5.20 pieces/100 g in 0-300 mm soil layer,which is their main enrichment area.However,the average abundance of microplastics decreases to 11.20±1.10 pieces/100 g in 400-800 mm soil depth.Moreover,the average particle size of microplastics is also linearly negatively correlated with soil depth.Microplastics with smaller particle size are easier to migrate as they pass through soil pores under the action of water and fertilizer.The research can provide s reference for understanding plastic mulch pollution.展开更多
To the Editor: Bacterial vaginosis (BV) is the most prevalent form ofvaginitis among women of reproductive age, affecting 8-23% of women globally. Clinically, BV is typically diagnosed using Amsel's criteria and t...To the Editor: Bacterial vaginosis (BV) is the most prevalent form ofvaginitis among women of reproductive age, affecting 8-23% of women globally. Clinically, BV is typically diagnosed using Amsel's criteria and the Nugent scoring system. However, these methods are inaccurate in many cases. In this study, we explored the feasibility of using length-heterogeneity-polymerase chain reaction (LH-PCR) for diagnosis of BV.展开更多
基金supported by the National Key R&D Program of China(2021YFC2600400)the Technology Innovation Program of the Chinese Academy of Agricultural Sciences(caascx-2017-2022-IAS)the Key R&D Program of Yunnan Province,China(202103AF140007)。
文摘Invasive alien ants(IAAs)are among the most aggressive,competitive,and widespread invasive alien species(IAS)worldwide.Wasmannia auropunctata,the greatest IAAs threat in the Pacific region and listed in“100 of the world’s worst IAS”,has established itself in many countries and on islands worldwide.Wild populations of W.auropunctata were recently reported in southeastern China,representing a tremendous potential threat to China’s agricultural,economic,environmental,public health,and social well-being.Estimating the potential geographical distribution(PGD)of W.auropunctata in China can illustrate areas that may potentially face invasion risk.Therefore,based on the global distribution records of W.auropunctata and bioclimatic variables,we predicted the geographical distribution pattern of W.auropunctata in China under the effects of climate change using an ensemble model(EM).Our findings showed that artificial neural network(ANN),flexible discriminant analysis(FDA),gradient boosting model(GBM),Random Forest(RF)were more accurate than categorical regression tree analysis(CTA),generalized linear model(GLM),maximum entropy model(MaxEnt)and surface distance envelope(SRE).The mean TSS values of ANN,FDA,GBM,and RF were 0.820,0.810,0.843,and 0.857,respectively,and the mean AUC values were 0.946,0.954,0.968,and 0.979,respectively.The mean TSS and AUC values of EM were 0.882 and 0.972,respectively,indicating that the prediction results with EM were more reliable than those with the single model.The PGD of W.auropunctata in China is mainly located in southern China under current and future climate change.Under climate change,the PGD of W.auropunctata in China will expand to higher-latitude areas.The annual temperature range(bio7)and mean temperature of the warmest quarter(bio10)were the most significant variables affecting the PGD of W.auropunctata in China.The PGD of W.auropunctata in China was mainly attributed to temperature variables,such as the annual temperature range(bio7)and the mean temperature of the warmest quarter(bio10).The populations of W.auropunctata in southern China have broad potential invasion areas.Developing strategies for the early warning,monitoring,prevention,and control of W.auropunctata in southern China requires more attention.
基金Supported by China Clean Development Mechanism Project(2014101)。
文摘Agrometeorological disasters severely impact agriculture in Heilongjiang Province.Flood is one of the main agrometeorological disasters in Heilongjiang Province.The temporal change in flood events in Heilongjiang Province from 1986to 2015 was studied using Mann-Kendall and Morlet wavelet methods,respectively.The results of Mann-Kendall analysis showed that the disaster rates of flood gradually stabilized from 1986 to 2015 with a confidence level of 99%.The Morlet wavelet variance analysis revealed that disaster rates of flood changed periodically at time scales of 3a,7a and 18a in Heilongjiang Province during1986-2015.The dominant period of the variation of flood disaster rate was about 18a over the past 30 years.The flood disaster rates were indicated in a positive phase during the period of 2016-2020 by the fitting curve of Morlet wavelet analysis.The annual average flood disaster indexes of single station,during 1986-2015 years were calculated,according to the precipitation data at 31 stations in Heilongjiang Province and the GIS software was used to analyze the spatial change in flood disasters in Heilongjiang Province from1986-2015.The results demonstrated that the southwest area of Heilongjiang Province was highly hazardous region of flood.The flood indices in the northern part of Songnen Plain and southwest of Heilongjiang Province presented the increment trends.
基金financially supported by the Key S&T Special Project of Yunnan Province (202202AE090034)the National Natural Science Foundation of China (42107410)+3 种基金the Science and Technology Project of Hebei Education Department (BJ2021026)the Hebei Financial Aid Program for Introduced Overseas Scholars (C20200330)the Taishan Industry Leading Talents High-Efficiency Agriculture Innovation Project (LJNY202125)the Expert Workstation of Dali
文摘Agricultural non-point source pollution is increasingly an important issue affecting surface water quality.Currently,the majority of the studies on nitrogen loss have focused on the agricultural field scale,however,the response of surface water quality at the watershed scale into the nitrogen loss at the field scale is poorly understood.The present study systematically reviewed the critical processes and major factors that nitrogen transport from farm fields to surface water bodies.The critical processes of farmland nitrogen entering surface water bodies involve the processes of nitrogen transport from farmland to ditches and the transformation processes of nitrogen during migration in ditches/rivers.Nitrogen transport from farmland to ditches is one of the prerequisites and critical processes for farmland nitrogen transport to surface water bodies.The transformation of nitrogen forms in ditches/rivers is an intermediate process in the migration of nitrogen from farmland to surface water bodies.Nitrogen loss from farmland is related to soil storage and exogenous inputs.Therefore,nitrogen input management should not only consider the current input,but also the contribution of soil storage due to the historical surpluses.Ditches/rivers have a strong retention capacity for nitrogen,which will significantly affect the process of farmland nitrogen entering surface water bodies.The factors affecting nitrogen transformation in river/ditches can be placed in four categories:(1)factors affecting hydraulic retention time,(2)factors affecting contact area,(3)factors affecting biological activity,and(4)forms and amount of nitrogen loading to river/ditches.Ditch systems are more biologically(including plants and microbes)active than rivers with biological factors having a greater influence on nitrogen transformation.When developing pollution prevention and control strategies,ecological ditches can be constructed to increase biological activity and reduce the amount of surplus nitrogen entering the water body.The present research should be valuable for the evaluation of environment impacts of nitrogen loss and the non-point source pollution control.
基金jointly sponsored by the National Key Technology Research and Development Program of China(No.2015BAD05B04)the Natural Science Foundation of Jiangxi Province,China(No.20202BAB215016)+3 种基金the Foundation of Jiangxi Educational Committee,China(No.GJJ191707)the Science and Technology Service Network Program of Chinese Academy of Sciences(STS Program)the Regional Soil Pollution Control Program of the Ministry of Agriculture and Rural Affairs of Chinathe Agro-Environmental Protection Program of Jiangxi Province,China。
文摘Cadmium(Cd)contamination in paddy soils means that the rice produced there may be unsafe for human consumption.A hydroponic study was conducted to enrich rice seedlings with zinc(Zn)or manganese(Mn),and the uptake and transport characteristics of Cd in these Zn-and Mn-rich seedlings were subsequently investigated using a greenhouse pot trial.The results showed that hydroponic cultivation in 10-50μmol L^(-1) Zn(ZnSO_(4)·7 H_(2)O)or50-250μmol L^(-1) Mn(MnSO_(4)·H_(2)O)for 30 d had no significant impact on rice growth,while the accumulation of Zn and Mn was 7.31-18.5 and 25.4-47.7 times higher,respectively,than in the control(no Zn or Mn addition).The accumulation of Cd in the Zn-and Mn-rich rice plants was 26.3%-38.6% and 34.4%-44.5% lower than that in the control,respectively,and the translocation factors of Cd from roots to shoots also decreased by 23.3%-41.3% and 25.3%-37.0%,respectively,after transplanting to Cd-contaminated soils.Furthermore,the relative expression levels of OsIRT1(Oryza sativa iron-regulated transporter 1)were downregulated by 40.1%-59.3% and 16.0%-25.9%,respectively,in the Zn-and Mn-rich seedling roots.This downregulation may indicate a possible mechanism contributing to the reductions in Cd absorption.Field experiments confirmed that the Zn-and Mn-rich seedlings produced brown rice(unpolished rice grains)with significantly decreased concentrations of Cd(34.2%-44.4%).This study provides an innovative method for reducing the food safety risks from rice grown on slightly to moderately Cd-contaminated paddy soils.
基金This work was financially supported by the Public Welfare Industry(Agriculture)Research Funding Project(Grant No.201303091).
文摘The management of livestock waste is an effective way to achieve emission reduction and carbon fixation in agriculture and rural areas.At present,aerobic composting and anaerobic fermentation are widely used in livestock waste treatment technology.In this study,pig manure management was taken as an example,a comprehensive environmental load index was constructed to quantitatively evaluate the environmental impacts of global warming,environmental acidification,eutrophication,and photochemical ozone synthesis during aerobic composting and anaerobic fermentation based on the life cycle assessment.The results showed that the potential values of aerobic composting and anaerobic fermentation were similar,and the order was global warming,environmental acidification,eutrophication,and photochemical ozone synthesis.Anaerobic fermentation contributed more to global warming,while aerobic composting contributed more to environmental acidification,eutrophication,and photochemical ozone synthesis.In addition,the environmental load index of aerobic composting was significantly higher than that of anaerobic fermentation.There were certainly regional differences in the environmental load index,and the environmental impact effect of anaerobic fermentation was low and more environmentally friendly.These findings provided a technical basis for livestock manure management in different regions of China,which was conducive to promoting animal husbandry emission reduction and carbon sequestration.
基金supported by the National Natural Science Foundation of China(grant numbers:32088102,32061123002,and 32101529)National Key Research and Development Program of China(grant number:2021YFC2600100)the Program of the Natural Science Foundation of Hebei Province of China(grant number:C2019204244).
文摘Semiochemical-based management strategies are important for controlling bark beetles,such as invasive Red Turpentine Beetle(Denroctonus valens),the causal agent for mass mortality of pine trees(Pinus spp.)in China.It has been previously shown that the pheromone verbenone regulates the attack density of this beetle in a dose-dependent manner and that the gut bacteria of D.valens are involved in verbenone production.However,molecular functional verification of the role of gut bacteria in the pheromone production of D.valens is still lacking.To better understand the molecular function of gut bacterial verbenone production,we chose a facultative anaerobic gut bacterium(Enterobacter xiangfangensis)of D.valens based on its strong ability to convert cis-verbenol to verbenone,as shown in our previous study,and investigated its transcriptomics in the presence or absence of cis-verbenol under anaerobic conditions(simulating the anoxic environment in the beetle's gut).Based on this transcriptome analysis,aldehyde dehydrogenase(ALDH1)was identified as a putative key gene responsible for verbenone production and was knocked-down by homologous recombination to obtain a mutant E.xiangfangensis strain.Our results show that these mutants had significantly decreased the ability to convert the monoterpene precursor to verbenone compared with the wild-type bacteria,indicating that ALDH1 is primarily responsible for verbenone conversion for this bacterium species.These findings provide further mechanistic evidence of bacterially mediated pheromone production by D.valens,add new perspective for functional studies of gut bacteria in general,and may aid the development of new gene silencing-based pest management strategies.
基金This work was supported by National Natural Science Foundation of China(Grant No.32060288)the Key R&D Projects of Xinjiang Production and Construction Corps(Grant No.2019AB007)the Special Project for Agricultural Ecological Environment Protection of Ministry of Agriculture and Rural Areas of China(2110402).
文摘In order to study the distribution of soil microplastics in the plastic film mulched farmland,the fluidization-centrifugation secondary density flotation method was optimized and improved to obtain the samples.The main components of microplastics were analyzed by Fourier spectrometer,and the surface morphology,porosity,particle size and abundance distribution characteristics of microplastics were studied by electron microscopy.The results showed that plastic mulch is the main source of microplastics.Its morphology mainly consists of fragments,fibers and particles.The size of debris microplastics was larger,with an average of 1.6300 mm,mainly distributed in the 0-300 mm cultivation layer while the size of particle micro plastic was smaller,with an average of 0.1400 mm.The width of fiber microplastic was 5-20μm,but the length could reach 0.2000-2.0000 mm,with an average particle size of 0.9200 mm.Fiber and granular microplastics could be seen in each soil layer.The surface layer of microplastics has a large number of pores characterized with a length of 50μm and a width of about 5-10μm,which might be an easy oxidation site for microplastics.This oxidation is continuous,making microplastics continuously decompose into smaller particles.The abundance of microplastics is negatively correlated with soil depth.The average abundance of microplastics is 161.50±5.20 pieces/100 g in 0-300 mm soil layer,which is their main enrichment area.However,the average abundance of microplastics decreases to 11.20±1.10 pieces/100 g in 400-800 mm soil depth.Moreover,the average particle size of microplastics is also linearly negatively correlated with soil depth.Microplastics with smaller particle size are easier to migrate as they pass through soil pores under the action of water and fertilizer.The research can provide s reference for understanding plastic mulch pollution.
基金This work was supported by grants from the National Natural Science Foundation of China (No. 81200411) and the Specialized Research Fund for the Doctoral Program of Higher Education Project of the New Teacher (No. 20120001120037).
文摘To the Editor: Bacterial vaginosis (BV) is the most prevalent form ofvaginitis among women of reproductive age, affecting 8-23% of women globally. Clinically, BV is typically diagnosed using Amsel's criteria and the Nugent scoring system. However, these methods are inaccurate in many cases. In this study, we explored the feasibility of using length-heterogeneity-polymerase chain reaction (LH-PCR) for diagnosis of BV.