Molecular characterization of a novel victorivirus isolated from the phytopathogenic fungus Phaeobotryon rhois.

Phaeobotryon rhois is an important pathogenic fungus that causes dieback and canker disease of woody hosts. A novel mycovirus, tentatively named "Phaeobotryon rhois victorivirus 1" (PrVV1), was identified in P. rhois strain SX8-4. The PrVV1 has a double-stranded RNA (dsRNA) genome that is 5,224 base pairs long and contains two open reading frames (ORF1 and ORF2), which overlap at a AUGA sequence. ORF1 encodes a polypeptide of 786 amino acids (aa) that contains the conserved coat protein (CP) domain of victoriviruses, while ORF2, encodes a large polypeptide of 826 aa that contains the conserved RNA-dependent RNA polymerase (RdRp) domain of victoriviruses. Our analysis of genomic structure, homology, and phylogeny indicated that PrVV1 is a novel member of the genus Victorivirus in the family Totiviridae. This is the first report of the complete genome sequence of a victorivirus that infects P. rhois.

Historical records reveal the distinctive associations of human disturbance and extreme climate change with local extinction of mammals.

Accelerated anthropogenic impacts and climatic changes are widely considered to be responsible for unprecedented species extinction. However, determining their effects on extinction is challenging owing to the lack of long-term data with high spatial and temporal resolution. In this study, using historical occurrence records of 11 medium- to large-sized mammal species or groups of species in China from 905 BC to AD 2006, we quantified the distinctive associations of anthropogenic stressors (represented by cropland coverage and human population density) and climatic stressors (represented by air temperature) with the local extinction of these mammals. We found that both intensified human disturbances and extreme climate change were associated with the increased local extinction of the study mammals. In the cold phase (the premodern period of China), climate cooling was positively associated with increased local extinction, while in the warm phase (the modern period) global warming was associated with increased local extinction. Interactive effects between human disturbance and temperature change with the local extinction of elephants, rhinos, pandas, and water deer were found. Large-sized mammals, such as elephants, rhinos, and pandas, showed earlier and larger population declines than small-sized ones. The local extinction sensitivities of these mammals to the human population density and standardized temperature were estimated during 1700 to 2000. The quantitative evidence for anthropogenic and climatic associations with mammalian extinction provided insights into the driving processes of species extinction, which has important implications for biodiversity conservation under accelerating global changes.

Transmission rate and control efficiency of COVID-19 was lower in warm and wet climate.

COVID-19 has caused huge damage to public health around the world, revealing the influencing factors are essential to take effective control. By using a global dataset covering 617 time series over the world, we estimated the transmission parameters and modeled human and climate effects on COVID-19 transmission. We found that the average transmission rate was lower in warm climate over the world and in wet climate (more precipitation) in Europe. The maximum transmission rate was lower in warm climate in the world, China and USA, and in wet climate in China. The control efficiency in the world, China, and USA was lower in warm and wet condition. In general, our results indicate that warm and wet climate do not favor transmission and human intervention of COVID-19, and COVID-19 transmission rate would be lower in warm and wet seasons or regions than in dry and cold ones.

Molecular characterization of a novel partitivirus isolated from the phytopathogenic fungus Aplosporella javeedii.

Aplosporella javeedii is a pathogenic fungus that causes canker and dieback of jujube in China. In this study, we report a new mycovirus, Aplosporella javeedii partitivirus 1 (AjPV1), isolated from A. javeedii strain NX55-3. The AjPV1 genome contains two double-stranded RNA elements (dsRNA1 and dsRNA2). The size of dsRNA1 is 2,360 bp, and it encodes a putative RNA-dependent RNA polymerase (RdRp), while dsRNA2 is 2,301 bp in length and encodes a putative capsid protein (CP). The sequences of RdRp and CP have significant similarity to those of members of the family Partitiviridae. Sequence alignment and phylogenetic analysis showed that AjPV1 is a new member of the family Partitiviridae that is related to members of the genus Betapartitivirus. To our knowledge, AjPV1 is the first mycovirus reported from A. javeedii.

Modeling analysis reveals the transmission trend of COVID-19 and control efficiency of human intervention.

BACKGROUND: A novel coronavirus disease (COVID-19) has caused huge damage to public health around the world. Revealing the transmission dynamics of COVID-19 and control efficiency is important for containing the spread of the virus. METHODS: By using a logistic growth model, we estimated the transmission parameters of COVID-19 in China and six other countries (Republic of Korea, Iran, Italy, Spain, France and Germany). The transmission parameters represent the maximum daily increase rate in the early stages of the epidemic and the control efficiency under human intervention. The control efficiency was determined by the significant decrease of the daily increase rate in time and cumulative cases. RESULTS: We found the daily increase rate of cumulative cases of COVID-19 decreased significantly in both time and cumulative cases in all countries, but the decreasing trend was not further reduced in other countries except for China and Republic of Korea. The response of the daily increase rate to control measures was much earlier than the number of new cases. CONCLUSIONS: Our results suggested that lockdown at the epicenter and social distancing effectively reduced the spread of COVID-19 in the early stage, but identification and isolation of patients, suspected cases and people with close contact at a community level is essential in further reduction of the daily increase rate of COVID-19.

Climate warming and humans played different roles in triggering Late Quaternary extinctions in east and west Eurasia.

Climate change and humans are proposed as the two key drivers of total extinction of many large mammals in the Late Pleistocene and Early Holocene, but disentangling their relative roles remains challenging owing to a lack of quantitative evaluation of human impact and climate-driven distribution changes on the extinctions of these large mammals in a continuous temporal-spatial dimension. Here, our analyses showed that temperature change had significant effects on mammoth (genus Mammuthus), rhinoceros (Rhinocerotidae), horse (Equidae) and deer (Cervidae). Rapid global warming was the predominant factor driving the total extinction of mammoths and rhinos in frigid zones from the Late Pleistocene and Early Holocene. Humans showed significant, negative effects on extirpations of the four mammalian taxa, and were the predominant factor causing the extinction or major extirpations of rhinos and horses. Deer survived both rapid climate warming and extensive human impacts. Our study indicates that both the current rates of warming and range shifts of species are much faster than those from the Late Pleistocene to Holocene. Our results provide new insight into the extinction of Late Quaternary megafauna by demonstrating taxon-, period- and region-specific differences in extinction drivers of climate change and human disturbances, and some implications about the extinction risk of animals by recent and ongoing climate warming.

Impacts of top predators and humans on the mammal communities of recovering temperate forest regions.

Top predators are important drivers in shaping ecological community structure via top-down effects. However, the ecological consequences and mechanisms of top predator loss under accelerated human impacts have rarely been quantitatively assessed due to the limited availability of long-term community data. With increases in top predator populations in northern China over the past two decades, forests with varying densities of top predators and humans provide an opportunity to study their ecological effects on mammal communities. We hypothesized a priori of conceptual models and tested these using structural equation models (SEMs) with multi-year camera trap data, aiming to reveal the underlying independent ecological effects of top predators (tigers, bears, and leopards) and humans on mammal communities. We used random forest models and correlations among species pairs to validate results. We found that top predator reduction could be related to augmented populations of large ungulates ("large ungulate release") and mesopredators ("mesopredator release"), consistent with observations of mammal communities in other ecosystems. Additionally, top predator reduction could be related to reduced small mammal abundance. Hierarchical SEMs identified three bottom-up pathways from forest quality to human activities, large ungulates, and some small mammals, and five top-down pathways from human activities and top predators to some small mammals, large ungulates, and mesopredators. Furthermore, our results suggest that humans showed predominant top-down effects on multiple functional groups, partially replacing the role of top predators, rather than be mediated by them; effects of humans and top predators appeared largely independent. Effects of humans on top predators were non-significant. This study provides novel insights into the effects of top predators and humans as super-predators on mammal communities in forest ecosystems and presents cues of bottom-up effects that can be translated into actionable management plans for improving forest quality, thereby supporting top predator recovery and work/life activities of local people.

Effects of Group Size on Behavior, Reproduction, and mRNA Expression in Brains of Brandt's Voles.

For social animals, a moderate group size is greatly important to maintain their reproductive success. However, the underlying neurobiological mechanism of group size on behavior and reproduction has rarely been investigated. In this study, we examined the effects of group size (1, 2, 4 pairs of adult male and female voles raised per cage) on behavior and reproduction. Meanwhile, the mRNA expression of stress and reproduction response-related genes in male brains was detected. We found that Brandt's voles (Lasiopodomys brandtii) in the large-sized group fight more severely than those in the small-sized group. Meanwhile, male voles were more anxious than females. The average number of embryos and litters per female in the medium-sized group was significantly higher than that of large-sized group. In male voles, stress- or reproduction-response mRNA expressions were more related to final group size or final density due to death caused by fighting. Our results indicated that a moderate group size was beneficial to the reproductive output of Brandt's voles. Our study highlights the combined effects of stress- or reproduction-related gene expression or behavior in regulating the fitness of voles with different group sizes.

Biological control of Fusarium crown rot of wheat with Chaetomium globosum 12XP1-2-3 and its effects on rhizosphere microorganisms.

Chaetomium globosum is a common plant endophytic fungi that exhibits great biocontrol potential in plant disease. Fusarium crown rot (FCR) is an important disease in wheat that seriously threatens wheat production worldwide. The control effect of C. globosum against wheat FCR remains unclear. In this study, we introduced an identified C. globosum 12XP1-2-3 and tested its biological control potential against wheat FCR. The hypha and fermentation broth exhibited an antagonistic effect against Fusarium pseudograminearum. Results from indoor experiments showed that C. globosum 12XP1-2-3 might delay the onset of symptoms of brown stem base and significantly reduced the disease index (37.3%). Field trials showed that wheat seeds coated with a spore suspension of 12XP1-2-3 grew better than the control seeds, had control effects of 25.9-73.1% on FCR disease, and increased wheat yield by 3.2-11.9%. Analysis of rhizosphere microorganisms revealed that seeds coated with C. globosum ('Cg' treatment) had a greater effect on fungal rather than on bacterial alpha diversity and may improve the health state of rhizosphere microorganisms, as reflected by the significantly increased fungal Shannon index at Feekes 11 and the increased complexity of the bacterial co-occurrence network but decreased complexity of the fungal network. Moreover, the accumulation of beneficial bacteria such as Bacillus and Rhizobium at Feekes 3, and Sphingomonas at Feekes 7 in the 'Cg' treatment may be the important contributions to healthier wheat growth state, significantly reduced relative abundance of Fusarium at Feekes 11, and reduced occurrence of FCR disease. These results provide a basis for further research on the mechanism of action of C. globosum and its application in the biological control of FCR in the field.

Synergistic effects of EP-1 and ivermectin mixture (iEP-1) to control rodents and their ectoparasites.

BACKGROUND: Ectoparasites of rodents play significant roles in disease transmission to humans. Conventional poisoning potentially reduces the population densities of rodents, however, they may increase the ectoparasite loads on the surviving hosts. EP-1 has been shown to have anti-fertility effects on many rodent species, while ivermectin is effective in controlling ectoparasites. In this study, we examined the combined effects of EP-1 and ivermectin mixture (iEP-1) baits on rodents and their corresponding flea/tick loads. RESULTS: In males, the weight of testis, epididymis, and seminiferous vesicle were reduced to less than 33%, 25%, and 17%, respectively, compared to the control group following administration of iEP-1 for 7 days. The weight of the uterus increased by approximately 75%. After 5 days of iEP-1 intake, all ticks were killed, whereas 94% of fleas on mice died after 3 days of bait intake. In the field test near Beijing, the flea index was reduced by more than 90% after 7 days of iEP-1 bait delivery. In a field test in Inner Mongolia, the weights of testis, epididymis, and seminiferous vesicle were significantly reduced by 27%, 32%, and 57%, respectively, 2 weeks after iEP-1 bait delivery. Approximately 36% rodents exhibited obvious uterine oedema accompanied by a weight increase of about 150%. The flea index was reduced by over 90%. CONCLUSION: Our results indicated that iEP-1 is a promising treatment for reducing the abundance of both small rodents and their ectoparasites; this will be effective for managing rodent damage and transmission of rodent-borne diseases associated with fleas and ticks. © 2022 Society of Chemical Industry.

Sustained population decline of rodents is linked to accelerated climate warming and human disturbance.

BACKGROUND: During the past three decades, sustained population decline or disappearance of cycles in small rodents have been observed. Both anthropogenic disturbance and climate warming are likely to be potential drivers of population decline, but quantitative analysis on their distinct effects is still lacking. RESULTS: Using time series monitoring of 115 populations (80 populations from 18 known rodent species, 35 mixed populations from unknown species) from 1980 in China (spanning 20-33 yrs), we analyzed association of human disturbances and climate warming with population dynamics of these rodent species. We found 54 of 115 populations showed a decreasing trend since 1980, and 16 of 115 showed an increasing trend. Human disturbances and climate warming showed significant positive associations with the population declines of most rodent species, and the population declines were more pronounced in habitats with more intensified human disturbance such as cities and farmlands or in high-latitude regions which experienced more increase of temperature. CONCLUSIONS: Our results indicate that the large-scale sustained population decline of small mammals in various ecosystems driven by the rapid increase of both climate warming and human disturbance is likely a signal of ecosystem dysfunction or transition. There is an urgent need to assess the risks of accelerated climate warming and human disturbance imposes on our ecosystems.

Factors influencing range contraction of a rodent herbivore in a steppe grassland over the past decades.

Climate warming and human disturbance are known to be key drivers in causing range contraction of many species, but quantitative assessment on their distinctive and interactive effects on local disappearance is still rare.In this study, we examined the association of climate warming and human disturbance stressors with local disappearance probability of Brandt's voles (Lasiopodomys brandtii) in a steppe grassland in northern China.We used logistic generalized additive models to quantify the relationship between local disappearance probability of Brandt's voles and environmental variables. The year following the last observation year was used to estimate the disappearance threshold of Brandt's voles. We projected the distribution change of Brandt's voles under future climate warming scenarios.We found climate warming attributed to local disappearance and range contraction for southern populations of Brandt's voles from 1971 to 2020. Human stressors and high vegetation coverage increased the probability of local disappearance of voles in years of abundant precipitation. The southern boundary retreated northward at a speed of 99.0 km per decade with the temperature rise of 0.36°C. The disappearance threshold of maximum air temperature of Brandt's voles in the warmest month (27.50 ± 1.61°C) was similar to the lower critical temperature of its thermal neutral zone.Our study suggests that the rapid climate change over the past decades contributed to the range contraction of its southern boundary of this keystone species in the steppe grassland of China. It is necessary to take actions to preserve the isolated populations of Brandt's voles from the effects of accelerated climate change and human disturbance.

Broad-scale climate variation drives the dynamics of animal populations: a global multi-taxa analysis.

Climate is a major extrinsic factor affecting the population dynamics of many organisms. The Broad-Scale Climate Hypothesis (BSCH) was proposed by Elton to explain the large-scale synchronous population cycles of animals, but the extent of support and whether it differs among taxa and geographical regions is unclear. We reviewed publications examining the relationship between the population dynamics of multiple taxa worldwide and the two most commonly used broad-scale climate indices, El Niño-Southern Oscillation (ENSO) and North Atlantic Oscillation (NAO). Our review and synthesis (based on 561 species from 221 papers) reveals that population changes of mammals, birds and insects are strongly affected by major oceanic shifts or irregular oceanic changes, particularly in ENSO- and NAO-influenced regions (Pacific and Atlantic, respectively), providing clear evidence supporting Elton's BSCH. Mammal and insect populations tended to increase during positive ENSO phases. Bird populations tended to increase in positive NAO phases. Some species showed dual associations with both positive and negative phases of the same climate index (ENSO or NAO). These findings indicate that some taxa or regions are more or less vulnerable to climate fluctuations and that some geographical areas show multiple weather effects related to ENSO or NAO phases. Beyond confirming that animal populations are influenced by broad-scale climate variation, we document extensive patterns of variation among taxa and observe that the direct biotic and abiotic mechanisms for these broad-scale climate factors affecting animal populations are very poorly understood. A practical implication of our research is that changes in ENSO or NAO can be used as early signals for pest management and wildlife conservation. We advocate integrative studies at both broad and local scales to unravel the omnipresent effects of climate on animal populations to help address the challenge of conserving biodiversity in this era of accelerated climate change.

Whole-genome sequencing reveals the genetic mechanisms of domestication in classical inbred mice.

BACKGROUND: The laboratory mouse was domesticated from the wild house mouse. Understanding the genetics underlying domestication in laboratory mice, especially in the widely used classical inbred mice, is vital for studies using mouse models. However, the genetic mechanism of laboratory mouse domestication remains unknown due to lack of adequate genomic sequences of wild mice. RESULTS: We analyze the genetic relationships by whole-genome resequencing of 36 wild mice and 36 inbred strains. All classical inbred mice cluster together distinctly from wild and wild-derived inbred mice. Using nucleotide diversity analysis, Fst, and XP-CLR, we identify 339 positively selected genes that are closely associated with nervous system function. Approximately one third of these positively selected genes are highly expressed in brain tissues, and genetic mouse models of 125 genes in the positively selected genes exhibit abnormal behavioral or nervous system phenotypes. These positively selected genes show a higher ratio of differential expression between wild and classical inbred mice compared with all genes, especially in the hippocampus and frontal lobe. Using a mutant mouse model, we find that the SNP rs27900929 (T>C) in gene Astn2 significantly reduces the tameness of mice and modifies the ratio of the two Astn2 (a/b) isoforms. CONCLUSION: Our study indicates that classical inbred mice experienced high selection pressure during domestication under laboratory conditions. The analysis shows the positively selected genes are closely associated with behavior and the nervous system in mice. Tameness may be related to the Astn2 mutation and regulated by the ratio of the two Astn2 (a/b) isoforms.

Climate change affected the spatio-temporal occurrence of disasters in China over the past five centuries.

Climate change may contribute to the spatio-temporal occurrence of disasters. Long-term studies of either homogeneous or heterogeneous responses of historical disasters to climate change are, however, limited by the quality and quantity of the available proxy data. Here we reconstruct spatio-temporal patterns of five types of disasters in China during the period AD 1368-1911. Our analyses of these time series reveal that warmer temperatures decreased the occurrence of disasters in the monsoon-affected parts of central-east China, but it increased the frequency and intensity of disasters along the boundary of arid and humid conditions in parts of southwest and northeast China, probably driven by the interplay among monsoon, westerlies, polar vortex and variation of temperature. Moreover, we show that drought and flood events had cascading effects on the occurrences of locust outbreaks, famine and human epidemics. Our findings suggest that climate can contribute to the spatio-temporal occurrence of disasters, and therefore may contribute to an improvement of China's regional to national risk management of future climate and environmental change.

Molecular Characterization of the First Alternavirus Identified in Fusarium oxysporum.

A novel mycovirus named Fusarium oxysporum alternavirus 1(FoAV1) was identified as infecting Fusarium oxysporum strain BH19, which was isolated from a fusarium wilt diseased stem of Lilium brownii. The genome of FoAV1 contains four double-stranded RNA (dsRNA) segments (dsRNA1, dsRNA 2, dsRNA 3 and dsRNA 4, with lengths of 3.3, 2.6, 2.3 and 1.8 kbp, respectively). Additionally, dsRNA1 encodes RNA-dependent RNA polymerase (RdRp), and dsRNA2- dsRNA3- and dsRNA4-encoded hypothetical proteins (ORF2, ORF3 and ORF4), respectively. A homology BLAST search, along with multiple alignments based on RdRp, ORF2 and ORF3 sequences, identified FoAV1 as a novel member of the proposed family "Alternaviridae". Evolutionary relation analyses indicated that FoAV1 may be related to alternaviruses, thus dividing the family "Alternaviridae" members into four clades. In addition, we determined that dsRNA4 was dispensable for replication and may be a satellite-like RNA of FoAV1-and could perhaps play a role in the evolution of alternaviruses. Our results provided evidence for potential genera establishment within the proposed family "Alternaviridae". Additionally, FoAV1 exhibited biological control of Fusarium wilt. Our results also laid the foundations for the further study of mycoviruses within the family "Alternaviridae", and provide a potential agent for the biocontrol of diseases caused by F. oxysporum.

Catechol amide derivatized polyhydroxylated fullerene as potential chelating agents of radionuclides: Synthesis, reactive oxygen species scavenging, and cytotoxic studies.

Radionuclide internal contamination can induce chemical and radioactive intoxication and produce harmful free radicals in vivo. At present, administration of chelating agents is the most effective treatment against nuclide contamination. However, traditional studies on chelating agents have ignored the damage caused by free radicals to the body. The present study aimed to develop a type of a bifunctional sequestering agent that can chelates nuclides and scavenges free radicals simultaneously. Therefore, a novel catechol amide-derivatized polyhydroxylated fullerene was designed and prepared. The poor water solubility of fullerene was ameliorated by chemically modifying hydrophilic catechol amide and multiple hydroxyl groups, and obtaining high water-soluble fullerene derivatives. The affinities of chelators were investigated via sulfochlorophenol competitive complexing method and antioxidant capacities were examined by electron paramagnetic resonance. The results revealed the good complexation of the designed and synthesized chelating agent with uranyl ions; and its efficiency in scavenging hydroxyl radicals. This chelating agent showed extremely low toxicity and notable protective effect against oxidative stress on A549 cells. Besides, in U(VI)-exposed A549 cells, immediate treatment with catechol amide-derivatized polyhydroxylated fullerene significantly decreased the lactate dehydrogenase (LDH) release by inhibiting the cellular U(VI) intake, promoting the intracellular U(VI) release and inhibiting the production of intracellular reactive oxygen species (ROS). These results suggest that this fullerene derivative may be a valuable in vivo antioxidant and radionuclide decorporation agent.

A Novel Ourmia-Like Mycovirus Confers Hypovirulence-Associated Traits on Fusarium oxysporum.

Fusarium wilt caused by Fusarium oxysporum f. sp. momordicae (FoM) is an important fungal disease that affects the production of bitter gourd. Hypovirulence-associated mycoviruses have great potential and application prospects for controlling the fungal disease. In this study, a novel ourmia-like virus, named Fusarium oxysporum ourmia-like virus 1 (FoOuLV1), was isolated from FoM strain HuN8. The viral genomic RNA is 2,712 nucleotides (nt) in length and contains an open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) using either standard or mitochondrial codes. In strain HuN8, there was also a FoOuLV1-associated RNA segment with 1,173 nt in length with no sequence homology. Phylogenetic analysis showed that FoOuLV1 is a member of the genus Magoulivirus of the family Botourmiaviridae. FoOuLV1 was found to be associated with hypovirulence in FoM. Moreover, FoOuLV1 and its hypovirulence trait can be transmitted horizontally to other FoM strains and also to other formae speciale strains of F. oxysporum. In addition, FoOuLV1 showed significant biological control effect against the bitter gourd Fusarium wilt. To our knowledge, this study reveals the first description of a hypovirulence-associated ourmia-like mycovirus, which has the potential to the biological control of Fusarium wilt.

Human plague system associated with rodent diversity and other environmental factors.

Plague remains a threat to public health and is considered as a re-emerging infectious disease today. Rodents play an important role as major hosts in plague persistence and driving plague outbreaks in natural foci; however, few studies have tested the association between host diversity in ecosystems and human plague risk. Here we use zero-inflated generalized additive models to examine the association of species richness with human plague presence (where plague outbreaks could occur) and intensity (the average number of annual human cases when they occurred) in China during the Third Pandemic. We also account for transportation network density, annual precipitation levels and human population size. We found rodent species richness, particularly of rodent plague hosts, is positively associated with the presence of human plague. Further investigation shows that species richness of both wild and commensal rodent plague hosts are positively correlated with the presence, but only the latter correlated with the intensity. Our results indicated a positive relationship between rodent diversity and human plague, which may provide suggestions for the plague surveillance system.

Quantifying the effects of climate and anthropogenic change on regional species loss in China.

Human-induced environmental and climate change are widely blamed for causing rapid global biodiversity loss, but direct estimation of the proportion of biodiversity lost at local or regional scales are still infrequent. This prevents us from quantifying the main and interactive effects of anthropogenic environmental and climate change on species loss. Here, we demonstrate that the estimated proportion of species loss of 252 key protected vertebrate species at a county level of China during the past half century was 27.2% for all taxa, 47.7% for mammals, 28.8% for amphibians and reptiles and 19.8% for birds. Both human population increase and species richness showed significant positive correlations with species loss of all taxa combined, mammals, birds, and amphibians and reptiles. Temperature increase was positively correlated with all-taxa and bird species loss. Precipitation increase was negatively correlated with species loss of birds. Human population change and species richness showed more significant interactions with the other correlates of species loss. High species richness regions had higher species loss under the drivers of human environmental and climate change than low-richness regions. Consequently, ongoing human environmental and climate changes are expected to perpetuate more negative effects on the survival of key vertebrate species, particularly in high-biodiversity regions.