Development of an on-site diagnostic LAMP assay for rapid differentiation of the invasive pest Phthorimaea absoluta (Meyrick) using insect tissues.

BACKGROUND: The tomato leafminer, Phthorimaea absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a destructive invasive pest that originated in South America and has spread within China since 2017. A rapid method for on-site identification of P. absoluta is urgently needed for interception of this pest across China. RESULTS: We developed a loop-mediated isothermal amplification (LAMP) technique to differentiate P. absoluta from Liriomyza sativae, Chromatomyia horticola, and Phthorimaea operculella using extracted genomic DNA, which was then refined to create an on-site LAMP diagnostic method that can be performed under field conditions without the need for laboratory equipment. CONCLUSION: In the present research, we developed an on-site diagnostic method for rapid differentiation of P. absoluta from other insects with similar morphology or damage characteristics in China. © 2024 Society of Chemical Industry.

Dual mutations in the whitefly nicotinic acetylcholine receptor ß1 subunit confer target-site resistance to multiple neonicotinoid insecticides.

Neonicotinoid insecticides, which target insect nicotinic acetylcholine receptors (nAChRs), have been widely and intensively used to control the whitefly, Bemisia tabaci, a highly damaging, globally distributed, crop pest. This has inevitably led to the emergence of populations with resistance to neonicotinoids. However, to date, there have been no reports of target-site resistance involving mutation of B. tabaci nAChR genes. Here we characterize the nAChR subunit gene family of B. tabaci and identify dual mutations (A58T&R79E) in one of these genes (BTß1) that confer resistance to multiple neonicotinoids. Transgenic D. melanogaster, where the native nAChR Dß1 was replaced with BTß1A58T&R79E, were significantly more resistant to neonicotinoids than flies where Dß1 were replaced with the wildtype BTß1 sequence, demonstrating the causal role of the mutations in resistance. The two mutations identified in this study replace two amino acids that are highly conserved in >200 insect species. Three-dimensional modelling suggests a molecular mechanism for this resistance, whereby A58T forms a hydrogen bond with the R79E side chain, which positions its negatively-charged carboxylate group to electrostatically repulse a neonicotinoid at the orthosteric site. Together these findings describe the first case of target-site resistance to neonicotinoids in B. tabaci and provide insight into the molecular determinants of neonicotinoid binding and selectivity.

Discovery of new triterpenoids from Leptopus clarkei and their antiproliferative activity on cancer cells.

Two new triterpenoids (1-2), along with six known analogues (3-8) were obtained from the dried whole plant of Leptopus clarkei. Compound 1 is a 3,4-seco-lupane-type triterpenoid, and compound 2 is a phenylpropanoid-conjugated pentacyclic triterpenoid possessing trans-p-coumaroyl unit attached to oleanane-type skeleton. This is the first report on chemical investigation of the L. clarkei, and the triterpenoid derivatives were found in this plant for the first time. The structures of the new compounds were unequivocally elucidated by HRESIMS and 1D/2D NMR data. Additionally, the isolated compounds were evaluated for theircytotoxicities against four cancer cell lines including HepG2, MCF-7, A549 and HeLa. Notably, compound 2 exhibited the most significant antiproliferative activity with IC50 less than 20 µM for four cancer lines.

The Genetic Diversity of White-Backed Planthoppers (Sogatella furcifera) between Myanmar and Yunnan Province of China.

In order to clarify the migration route and the source of white-backed planthopper (WBPH) (Sogatella furcifera) between Myanmar and Yunnan Province, China, we collected six populations throughout Myanmar and five populations around the border areas in Yunnan Province, China. A total of 790 base pairs in the mtDNA COI genes from 416 individuals were obtained. A total of 43 haplotypes were identified, among which 37 were unique haplotypes, and the remaining 6 were shared among different populations. Two common shared haplotypes (H_1 and H_2) had a widespread distribution in all populations and accounted for 88.8% of the total haplotype frequency, suggesting a high-level gene flow among the Myanmar and Yunnan populations. Bayesian skyline plot (BSP) analysis results indicated that the effective population size of WBPH expanded between about 10,000 and 7000 years ago, and S. furcifera might follow the post-LGM (Last Glacial Maximum) expansion pattern. Based on the total migrant (Nem) value, it can be deduced that north and northeast Myanmar were the primary migration sources for WBPH populations in the southwest and south Yunnan regions. This study aims to contribute to the sustainable regional management of this important rice pest and provide new insights into the genetic diversity of WBPH in Southeast Asia.

LncRNA HOTAIR Enhances Epithelial-to-mesenchymal Transition to Promote the Migration and Invasion of Liver Cancer by Regulating NUAK1 via Epigenetic Inhibition miR-145-5p Expression.

LncRNA HOTAIR play important roles in the epigenetic regulation of carcinogenesis and progression in liver cancer. Previous studies suggest that the overexpression of HOTAIR predicts poor prognosis. In this study, through transcriptome sequencing data and in vitro experiments, we found that HOTAIR were more highly expressed and there is significantly positive relationship between HOTAIR and NUAK1 in liver cancer tissues and cell lines. miR-145-5p was downregulated and showed negative correlation with HOTAIR and NUAK1. Transfect Sh-HOTAIR, LZRS-HOTAIR, miR-145 mimic, miR-145 inhibitor to change the expression of HOTAIR and miR-145-5p. The addition of HTH-01-015 inhibits the expression of NUAK1. HOTAIR knockdown, miR-145-5p upregulation and NUAK1 inhibition all repressed migration, invasion and metastasis and reversed the epithelial-to-mesenchymal transition in SNU-387 and HepG2 cells. We also showed that HOTAIR recruiting and binding PRC2 (EZH2) epigenetically represses miR-145-5p, which controls the target NUAK1, thus contributing to liver cancer cell-EMT process and accelerating tumor metastasis. Moreover, it is demonstrated that HOTAIR crosstalk with miR-145-5p/NUAK1 during epigenetic regulation. Our findings indicate that HOTAIR/miR-145-5p/NUAK1 axis acts as an EMT regulator and may be candidate prognostic biomarker and targets for new therapies in liver cancer.

Application of Microhaplotypes in Sibling Kinship Testing.

OBJECTIVES: To investigate the efficacy of different numbers of microhaplotype (MH) loci and the introduction of different reference samples on the identification of full sibling, half sibling and differentiation between full sibling and half sibling kinships, and to explore the effect of changing mutation rate on sibling testing. METHODS: First, a family map involving three generations was established, and four full sibling identification models, five half sibling identification models and five models distinguishing full and half siblings were constructed for different reference samples introduced. Based on the results of the previous study, two sets of nonbinary SNP-MH containing 34 and 54 loci were selected. Based on the above MH loci, 100 000 pairs of full sibling vs. unrelated individuals, 100 000 pairs of half sibling vs. unrelated individuals and 100 000 pairs of full sibling vs. half sibling were simulated based on the corresponding sibling kinship testing models, and the efficacy of each sibling kinship testing model was analyzed by the likelihood ratio algorithm under different thresholds. The mutant rate of 54 MH loci was changed to analyze the effect of mutation rate on sibling identification. RESULTS: In the same relationship testing model, the systematic efficacy of sibling testing was positively correlated with the number of MH loci detected. With the same number of MH loci, the efficacy of full sibling testing was better than that of uncle or grandfather when the reference sample introduced was a full sibling of A, but there was no significant difference in the identification efficacy of the four reference samples introduced for full sibling and half sibling differentiation testing. In addition, the mutation rate had a slight effect on the efficacy of sibling kinship testing. CONCLUSIONS: Increasing the number of MH loci and introducing reference samples of known relatives can increase the efficacy of full sibling testing, half sibling testing, and differentiation between full and half sibling kinships. The level of mutation rate in sibling testing by likelihood ratio method has a slight but insignificant effect on the efficacy.

Pyrifluquinazon baseline susceptibility and inhibition of Tomato chlorosis virus transmission by Bemisia tabaci.

BACKGROUND: Tomato chlorosis virus (ToCV) is associated with tomato yellow leaf disorder diseases in more than 20 countries. ToCV can be transmitted in a semipersistent manner by whitefly vectors such as Bemisia tabaci. Controlling the vector pests by using chemical insecticides is an efficient and effective approach to reduce and interrupt the virus transmission. Pyrifluquinazon is a new pyridine azomethine derivative, showing insecticidal toxicity to sucking pests by disturbing their feeding behavior. However, limited attention has been paid to the performance of pyrifluquinazon against B. tabaci and ToCV transmission. RESULT: This study showed the lethal concentration of 50% (LC50 ) values of pyrifluquinazon to 22 B. tabaci field populations ranged from 0.54 to 2.44 mg L-1 , and the baseline susceptibility of B. tabaci to pyrifluquinazon was 1.24 mg L-1 with a 95% confidence limit of 0.35-1.85 mg L-1 . Pyrifluquinazon and afidopyropen did not show cross-resistance to dinotefuran and pymetrozine in B. tabaci, which both inhibited the feeding activities of B. tabaci. The antifeedant concentration of 50% (AFC50 ) values at 48 h were 0.70 mg L-1 for pyrifluquinazon and 2.13 mg L-1 for afidopyropen. Foliar application of pyrifluquinazon and afidopyropen reduced the ToCV transmission by 40.91% and 33.33%, respectively and significantly decreased the ToCV loads in tomato plants under laboratory conditions. CONCLUSION: These results provided new information about the effects of modulators of the vanilloid-type transient receptor potential channel on the toxicity to B. tabaci and inhibition of ToCV transmission. © 2023 Society of Chemical Industry.

Heterogeneous distribution of Cardinium in whitefly populations is associated with host nuclear genetic background.

Inherited bacterial symbionts are very common in arthropods, but infection frequency can vary widely among populations. Experiments and interpopulation comparisons suggest that host genetic background might be important in explaining this variation. Our extensive field investigation showed that the infection pattern of the facultative symbiont Cardinium was heterogeneous across geographical populations of the invasive whitefly Bemisia tabaci Mediterranean (MED) in China, with genetic nuclear differences evident in 2 of the populations: 1 with a low infection rate (SD line) and 1 with a high infection rate (HaN line). However, whether the heterogeneous frequency of Cardinium is associated with the host genetic background remains poorly understood. Here, we compared the fitness of the Cardinium-infected and uninfected sublines with similar nuclear genetic backgrounds from SD and HaN lines, respectively, and further determine whether host extranuclear or nuclear genotype influenced the Cardinium-host phenotype by performing 2 new introgression series of 6 generations between SD and HaN lines (i.e., Cardinium-infected females of SD were backcrossed with uninfected males of HaN, and vice versa). The results showed that Cardinium provides marginal fitness benefits in the SD line, whereas Cardinium provides strong fitness benefits in the HaN line. Further, both Cardinium and the Cardinium-host nuclear interaction influence the fecundity and pre-adult survival rate of B. tabaci, whereas the extranuclear genotype does not. In conclusion, our results provide evidence that Cardinium-mediated fitness effects were closely associated with the host genetic background, which provides a fundamental basis for understanding the mechanism underlying the heterogeneous distribution of Cardinium in B. tabaci MED populations across China.

Novel Avian Influenza Virus (H5N1) Clade 2.3.4.4b Reassortants in Migratory Birds, China.

Two novel reassortant highly pathogenic avian influenza viruses (H5N1) clade 2.3.4.4b.2 were identified in dead migratory birds in China in November 2021. The viruses probably evolved among wild birds through different flyways connecting Europe and Asia. Their low antigenic reaction to vaccine antiserum indicates high risks to poultry and to public health.

Ecological Factors Associated with the Distribution of Bemisia tabaci Cryptic Species and Their Facultative Endosymbionts.

The sweetpotato whitefly, Bemisia tabaci species complex, comprises at least 44 morphologically indistinguishable cryptic species, whose endosymbiont infection patterns often varied at the spatial and temporal dimension. However, the effects of ecological factors (e.g., climatic or geographical factors) on the distribution of whitefly and the infection frequencies of their endosymbionts have not been fully elucidated. We, here, analyzed the associations between ecological factors and the distribution of whitefly and their three facultative endosymbionts (Candidatus Cardinium hertigii, Candidatus Hamiltonella defensa, and Rickettsia sp.) by screening 665 individuals collected from 29 geographical localities across China. The study identified eight B. tabaci species via mitochondrial cytochrome oxidase I (mtCOI) gene sequence alignment: two invasive species, MED (66.9%) and MEAM1 (12.2%), and six native cryptic species (20.9%), which differed in distribution patterns, ecological niches, and high suitability areas. The infection frequencies of the three endosymbionts in different cryptic species were distinct and multiple infections were relatively common in B. tabaci MED populations. Furthermore, the annual mean temperature positively affected Cardinium sp. and Rickettsia sp. infection frequencies in B. tabaci MED but negatively affected the quantitative distribution of B. tabaci MED, which indicates that Cardinium sp. and Rickettsia sp. maybe play a crucial role in the thermotolerance of B. tabaci MED, although the host whitefly per se exhibits no resistance to high temperature. Our findings revealed the complex effects of ecological factors on the expansion of the invasive whitefly.

Genome-Wide Identification and Expression Analysis of the Cytochrome P450 Gene Family in Bemisia tabaci MED and Their Roles in the Insecticide Resistance.

The whitefly, Bemisia tabaci MED (Hemiptera: Aleyrodidae), is an omnivorous agricultural pest, which causes huge economic losses to agriculture and is highly resistant to many pesticides. The overexpression of cytochrome P450 may play an important role in host adaptation and insecticide resistance in B. tabaci MED. Therefore, the present study systematically analyzed the cytochrome P450 gene family at the genome-wide level to understand its function in B. tabaci MED. Our analysis identified 58 cytochrome P450 genes in B. tabaci MED, among which 24 were novel. Phylogenetic analysis revealed broad functional and species-specific diversification in B. tabaci MED P450, suggesting the role of multiple P450 genes in detoxifying. Reverse transcription-real time quantitative PCR (RT-qPCR) showed that CYP4CS2, CYP4CS5, CYP4CS6, CYP4CS8, CYP6DW4, CYP6DW5, CYP6DW6, CYP6DZ8, and CYP6EN1 genes increased significantly after two days of exposure to imidacloprid. Interestingly, all nine genes belonged to the CYP4 and CYP6 families. A decrease in the expression of five genes (CYP6DW4, CYP6DW5, CYP6DW6, CYP6DZ8, and CYP4CS6) via RNA interference (RNAi) resulted in a significant increase in the mortalities of whiteflies when exposed to imidacloprid. These results indicate that the overexpression of the P450 genes may play an essential role in imidacloprid tolerance of B. tabaci MED. Thus, the present study provides basic information on P450 genes in B. tabaci MED, which will further help elucidate the insecticide resistance mechanism in the agricultural pest whitefly.

Prevalence, evolution, replication and transmission of H3N8 avian influenza viruses isolated from migratory birds in eastern China from 2017 to 2021.

The continued evolution and emergence of novel influenza viruses in wild and domestic animals poses an increasing public health risk. Two human cases of H3N8 avian influenza virus infection in China in 2022 have caused public concern regarding the risk of transmission between birds and humans. However, the prevalence of H3N8 avian influenza viruses in their natural reservoirs and their biological characteristics are largely unknown. To elucidate the potential threat of H3N8 viruses, we analyzed five years of surveillance data obtained from an important wetland region in eastern China and evaluated the evolutionary and biological characteristics of 21 H3N8 viruses isolated from 15,899 migratory bird samples between 2017 and 2021. Genetic and phylogenetic analyses showed that the H3N8 viruses circulating in migratory birds and ducks have evolved into different branches and have undergone complicated reassortment with viruses in waterfowl. The 21 viruses belonged to 12 genotypes, and some strains induced body weight loss and pneumonia in mice. All the tested H3N8 viruses preferentially bind to avian-type receptors, although they have acquired the ability to bind human-type receptors. Infection studies in ducks, chickens and pigeons demonstrated that the currently circulating H3N8 viruses in migratory birds have a high possibility of infecting domestic waterfowl and a low possibility of infecting chickens and pigeons. Our findings imply that circulating H3N8 viruses in migratory birds continue to evolve and pose a high infection risk in domestic ducks. These results further emphasize the importance of avian influenza surveillance at the wild bird and poultry interface.

Infection density pattern of Cardinium affects the responses of bacterial communities in an invasive whitefly under heat conditions.

Communities of bacteria, especially symbionts, are vital for the growth and development of insects and other arthropods, including Bemisia tabaci Mediterranean (MED), a destructive and invasive insect pest. However, the infection density patterns and influence factors of bacteria in whiteflies, which mainly include symbionts, remain largely unclear. To reveal the different density patterns of Cardinium in B. tabaci MED populations and the impacts of high temperatures on whiteflies with different Cardinium density infection patterns, 2 isofemale lines isolated from B. tabaci MED from the same geographical population of China and from B. tabaci MED collected from other countries and locations were examined using several techniques and methods, including fluorescence in situ hybridization (FISH), quantitative real-time polymerase chain reaction (qPCR), 16S rRNA gene sequencing, and 2b-RAD sequencing. The results showed that there were 2 different infection density patterns of Cardinium in B. tabaci MED (including 1 high-density pattern and 1 low-density pattern). For whiteflies with low-density Cardinium, conventional PCR could not detect Cardinium, but the other techniques confirmed that there was a low level of Cardinium within hosts. High temperature significantly decreased the diversity of bacterial communities: the relative titer of Cardinium increased but the density of Rickettsia decreased in the isofemale line with high-density Cardinium. However, high temperature did not influence the diversity and symbiont density in the line with low-density Cardinium. Moreover, high temperature influenced the functions of bacterial communities in whiteflies with high-density Cardinium but did not affect the bacterial functions in whiteflies with low-density Cardinium. Our results provide novel insights into the complex associations between symbionts and host insects.

Resistance to dinotefuran in Bemisia tabaci in China: status and characteristics.

BACKGROUND: Bemisia tabaci (Gennadius) is a serious agricultural pest worldwide. Neonicotinoids are the most important new class of synthetic insecticides used in the management of B. tabaci. However, B. tabaci populations have developed resistance to various active ingredients in neonicotinoids following long-term and widespread application. RESULTS: Dinotefuran exhibited high toxicity against most B. tabaci field populations. One population (Din-R) with a high level of resistance to dinotefuran (255.6-fold) was first identified in the field. The Din-R population exhibited medium to high levels of resistance to all the tested neonicotinoid insecticides and a high level of resistance to spinetoram. Genetic inheritance analysis revealed that resistance to dinotefuran was incompletely recessive and polygenic. The synergist piperonyl butoxide significantly increased the toxicity of dinotefuran to Din-R. P450 activity in the Din-R population was 2.19-fold higher than in the susceptible population. RNA-sequencing analysis showed that 12 P450 genes were significantly upregulated in the Din-R population, of which CYP6DW5, CYP6JM1 and CYP306A1 were found to exhibit more than 3.00-fold higher expression in Din-R when using a reverse transcription quantitative real-time polymerase chain reaction. Expression of eight P450 genes was obviously induced by dinotefuran, and CYP6DW5 showed the highest expression level. After knockdown of CYP6DW5 in Din-R, the toxicity of dinotefuran increased significantly. CONCLUSION: P450 had a crucial role in dinotefuran resistance in B. tabaci, and CYP6DW5 was involved in the resistance. These results provide important information for the management of resistance in B. tabaci and improve our understanding of the resistance mechanism of dinotefuran. © 2022 Society of Chemical Industry.

Expression profile of CYP402C1 and its role in resistance to imidacloprid in the whitefly, Bemisia tabaci.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cosmopolitan insect pest causing serious damage to crop production. Cytochromes P450 (CYPs) of B. tabaci are widely known to be involved in the metabolic resistance to a variety of insecticides, continuously increasing the difficulty in controlling this pest. In this study, four P450 genes (CYP6CM1, CYP6CX1, CYP6CX3, and CYP402C1) in B. tabaci exhibited correlations with the resistance to imidacloprid. We have focused on trying to understand the function and metabolism capacity of CYP402C1. The expression profiles of CYP402C1 were examined by reverse transcription quantitative real-time PCR and fluorescence in situ hybridizations. Its role in resistance to imidacloprid was investigated by RNA interference, transgenic Drosophila melanogaster, and heterologous expression. The results showed that CYP402C1 was highly expressed in the active feeding stages of B. tabaci, such as nymphs and female adults. CYP402C1 was mainly expressed in midguts of nymphs and adults, especially in the filter chamber. Knockdown of CYP402C1 significantly decreased the resistance of B. tabaci to imidacloprid by 3.96-fold (50% lethal concentration: 186.46 versus 47.08 mg/L). Overexpression of CYP402C1 in a transgenic D. melanogaster line (Gal4 > UAS-CYP402C1) significantly increased the resistance to imidacloprid from 12.68- to 14.92-fold (129.01 and 151.80 mg/L versus 1925.14 mg/L). The heterologous expression of CYP402C1 showed a metabolism ability of imidacloprid (imidacloprid decreased by 12.51% within 2 h). This study provides new insights for CYP402C1 function in B. tabaci and will help develop new strategies in B. tabaci control and its insecticide resistance.

Cardinium infection alters cotton defense and detoxification metabolism of its whitefly host.

Field monitoring revealed that the infection ratio of the bacterial symbiont Cardinium in the whitefly (Bemisia tabaci MED) was relatively low in northern China. However, the role of this symbiont and the symbiont-whitefly-host plant interaction mechanism are poorly understood. We investigated the influence of Cardinium on the competitiveness of the host whitefly and the physiological interaction between the host plants and host whiteflies. Cardinium-infected whiteflies were displaced by uninfected whiteflies after 5 generations, which showed that Cardinium infection reduced whitefly competitiveness. The defense response genes of cotton significantly decreased under infestation by infected whiteflies compared to uninfected whiteflies. The expression of detoxification metabolism genes, especially the uridine 5'-diphospho-glucuronyltransferase and P450 genes, in infected whiteflies significantly decreased. These results demonstrated that Cardinium could inhibit the defense response of the host plant and decrease the detoxification metabolism ability of the host whitefly. The reduced competitiveness of infected whiteflies may be associated with the inhibition of the whitefly detoxification metabolism by Cardinium, resulting in the reduced performance of infected whiteflies. However, Cardinium infection can suppress plant defenses, which may benefit both infected and uninfected whiteflies when they coexist. This research illustrates the symbiont-whitefly-host plant interaction mechanism and the population dynamics of the whitefly.

Both symbionts and environmental factors contribute to shape the microbiota in a pest insect, Sogatella furcifera.

Introduction: Bacterial symbionts are prevalent in arthropods globally and play a vital role in the fitness and resistance of hosts. While several symbiont infections have been identified in the white-backed planthopper Sogatella furcifera, the impact of environmental factors on the microbiota within S. furcifera remains elusive. Methods: In this study, a total of 142 S. furcifera individuals from 18 populations were collected from 14 locations across six countries (China, Thailand, Myanmar, Cambodia, Vietnam, and Laos) analyzed with 2bRAD-M sequencing, to examine the effects of symbionts on the microbiota in the S. furcifera population, as well as the vital effects of environmental factors on the bacterial communities. Results and discussion: Based on the results, in S. furcifera, the presence of symbionts Wolbachia and Cardinium negatively influenced the abundance of other bacteria, including Enterobacter, Acinetobacter, and Lysinibacillus, while Wolbachia infection significantly decreased the diversity of the microbial community. Moreover, several environmental factors, including longitude, latitude, temperature, and precipitation, affected the abundance of symbionts and microbiota diversity in S. furcifera. These results collectively highlight the vital role of Wolbachia in S. furcifera microbiota, as well as the intricate effects of environmental factors on the bacterial communities of S. furcifera.

Predicting potential global distribution and risk regions for potato cyst nematodes (Globodera rostochiensis and Globodera pallida).

Potato cyst nematodes (PCNs), golden (yellow) cyst nematode (Globodera rostochiensis, gPCN) and pale (white) cyst nematode (G. pallida, pPCN), are important invasive pests in many countries and regions where they can cause significant yield and economic loss for agriculture. Prediction and identification of habitats suitable for PCNs are critical for developing biosecurity strategies, both pre and post border, to maximise the potential for early elimination should an incursion occur. To date, the potential global distribution of PCNs has not been thoroughly studied. Therefore, this study conducted a species distribution model to illustrate the potential global distribution of PCNs and risk regions. In this study, the Maximum Entropy Model (Maxent) associated with the Geographic Information System (GIS) was employed to reveal the potential distribution of the gPCN and pPCN. In addition to bioclimate, soil quality was also included in the model. The global cultivated lands, whether the susceptible hosts were present or not, were used to assess the maximum potential risk regions. The limitation factors for PCNs distribution were also assessed. Results showed that 66% of the global land surface was suitable for gPCN or pPCN or both, and both species can colonise more than 75% of the global cultivated lands. The coldest quarter's mean temperature and precipitation were critical limitations in unsuitable regions. In summary, the global risk maps of PCNs contribute valuable additional information that complements previous national/regional distribution predictions. The results of this distribution research will contribute practical support for decision-makers and practitioners to implement biosecurity strategies from a global perspective, that incorporate prevention or promptly enforce control practices to limit the damage caused by future incursions.