Transboundary Spread Research Articles

Developing Predictive Models and Early Warning Systems for Invading Pathogens: Wheat Rusts.

Innovations in aerobiological and epidemiological modeling are enabling the development of powerful techniques to infer connectivity networks for transboundary pathogens in ways that were not previously possible. The innovations are supported by improved access to historical and near real-time highly resolved weather data, multi-country disease surveillance data, and enhanced computing power. Using wheat rusts as an exemplar, we introduce a flexible modeling framework to identify characteristic pathways for long-distance spore dispersal within countries and beyond national borders. We show how the models are used for near real-time early warning systems to support smallholder farmers in East Africa and South Asia. Wheat rust pathogens are ideal exemplars because they continue to pose threats to food security, especially in regions of the world where resources for control are limited. The risks are exacerbated by the rapid appearance and spread of new pathogenic strains, prodigious spore production, and long-distance dispersal for transboundary and pandemic spread.

Prevalence of Bacterial Diseases in Ornamental Fish Rearing Units of Kolathur, Tamil Nadu, India

Background: Ornamental fishkeeping is a widely embraced hobby globally. In India, Kolathur, Chennai Tamil Nadu is an important ornamental fish hub involved in trading of ornamental fishes. As a lucrative industry, the ornamental fish trade faces significant challenges due to diseases caused by various pathogens, impacting market value and causing economic losses. Methods: A surveillance study was conducted in Kolathur, Chennai, from December 2022 to August 2023. Ornamental fish samples exhibiting clinical signs of bacterial diseases were collected from the farms and retail outlets. Bacterial isolation and identification techniques, including conventional and PCR detection, were employed to isolate and identify bacterial strains from infected fish samples. Result: Out of 196 fish samples, 51 (29.4%) were found infected with bacterial pathogens. The prevalent bacterial pathogens included Aeromonas veronii (47%), A.caviae (17%), Pseudomonas aeruginosa (15%), Streptococcus sp. (11.3%), Mycobacterium fortuitum (5.6%) and Shewanella sp. (3.7%). Different fish families exhibited varying susceptibility to bacterial diseases, with cyprinids being the most affected (56.6%). The findings stress the importance of adhering to health regulations to prevent transboundary spread and suggest the development of targeted management strategies to sustain the health of ornamental fish populations.

Single-Gene Phylogeny of Brucella Melitensis in Milk Samples of Ewes in Kalar District, Iraqi Kurdistan Region

The present study aimed to investigate the frequency rate of ovine brucellosis in Kalar district, Iraqi Kurdistan Region and to identify Brucella species in the milk of seropositive ewes via DNA sequencing and phylogenetic analyses. Blood samples were collected from 300 ewes and subjected to the Rose Bengal Plate Test (RBPT), and milk samples of the seropositive animals were collected and underwent a PCR assay using genus- and species-specific primers. Eventually, the positive PCR products were processed for nucleotide sequencing and single-gene phylogenetic analyses. The RBPT revealed a high percentage (11.33%) of seropositive ewes (34 out of 300), and the PCR assay revealed 35.3% (12/34) Brucella genus-positive DNA templates. At the species level, 58.3% (7/12) of templates were positive for the melitensis-specific primers and negative for the abortus and ovis-specific primers, whereas the other five were negative for all species-specific primers used in this study (abortus, melitensis and ovis). The nucleotides sequence of Brucella strains detected in this study showed a 99-100% homology with eight GenBank-published B. melitensis strains, and the single gene phylogenetic analysis showed a close relationship between these strains and the B. melitensis biovar 3 strain NI, China. The high percentage of seropositive ewes showed that brucellosis represents a major obstacle to the animals’ production improvement and a major risk to public health in the Iraqi Kurdistan Region and the single gene phylogenetic analysis confirms its transboundary spread and attracts attention for more strict control programs.

Characterization, histopathology and immunogenicity of the lumpy skin disease virus isolated during 2019-20 in Bangladesh.

Lumpy skin disease (LSD) is a highly contagious vector-borne viral disease of cattle. LSD has emerged in Bangladesh in 2019, causing significant economic losses due to its high morbidity and mortality. This research was designed to isolate, identify, and assess the immunogenicity of LSD virus (LSDV) using nodular tissue samples obtained from affected cattle during the 2019-20 outbreak across nine districts of Bangladesh. To determine the presence of LSDV in nodular tissues, we initially used iiPCR and PCR, followed by histopathological examination. 151 were positive via iiPCR and PCR among the 180 collected samples. The PCR positive 151 samples were then inoculated into 10-day-old embryonated chicken eggs via the CAM route to isolate LSDV, confirmed through PCR. Subsequently, partial sequencing and phylogenetic analysis of the P32 gene were performed to determine the origin of the circulating LSDV strain. The immunogenicity of selected LSDV strains was assessed through an ELISA test. The PCR results revealed a distinct positive band at 192 bp in both the nodular tissue samples and the LSDV isolated from chicken embryo inoculations. Microscopic analysis of the nodular lesions revealed thickening of the epidermis, ballooning degeneration of keratinocytes, and proliferation of follicular epithelia. Additionally, mononuclear infiltration was observed at the demarcation line between infected and healthy tissue, with necrosis of muscular tissues beneath the epidermis. The LSDV isolate from Bangladesh exhibited a close genetic relationship with LSDV strains isolated from neighboring and other regional countries including India, Myanmar, and Mongolia. This observation strongly suggests the possibility of a transboundary spread of the LSD outbreak in Bangladesh during 2019-2020. The results of the immunogenicity test showed that the serum antibody titer remained at a protective level for up to 18 months following secondary immunization with inactivated LSDV antigen. This finding suggests that the inactivated LSDV antigen could be a potential vaccine candidate to protect cattle in Bangladesh against LSDV. In conclusion, our research successfully isolated, identified, and characterized LSDV in cattle nodular tissues from the 2019-20 outbreak in Bangladesh. Furthermore, it provided insights into the probable origin of the circulating strain and investigated a potential vaccine candidate to protect cattle in the region from LSDV.

Isolation and molecular characterization of lumpy skin disease virus from Tamil Nadu, India during the outbreaks from 2020 to 2022.

Lumpy skin disease (LSD) caused by LSD virus is a WOAH notifiable, high-impact, transboundary poxviral disease of bovines. The first official report of LSDV in India is from Odisha state during August 2019. Since then, cases have been reported from many states including Tamil Nadu, a Southern state of India. The present study deals with isolation and molecular characterization of LSDV from Tamil Nadu during the period August 2020 to July 2022. LSDV was isolated in embryonated chicken eggs (ECE) and BHK 21 cells and was characterized based on P32, RPO30, and GPCR genes. The phylogenetic analysis revealed that Tamil Nadu isolates from India are closely related to other Indian strains, Kenyan strains and strains from neighboring countries such as Bangladesh, Nepal, and Myanmar confirming the common exotic source for the transboundary spread across borders. The presence of unique signature of amino acid (aa) at specific positions (A11, T12, T34, S99, and P199) in the GPCR sequence confirmed the identity of LSDV. A twelve nucleotide (nt94-105) insertion and corresponding aa (TILS) at 30-33 position was found in GPCR sequence and characteristic amino acid proline at 98 position (P98) in the RPO30 gene sequence of our isolates was similar to strains from Bangladesh, Nepal, and Myanmar. Further, dissimilarity of our isolates from Neethling like vaccine strains confirms the circulation of virulent filed strains responsible for the outbreaks.

Phytosanitary management of ICARDA’s germplasm seed collections for safe movement and better future use

Abstract Germplasm exchange from international genebanks and breeding programs is vital for successful crop improvement programs. More than 10,000 different accessions of wheat, barley, lentil, faba bean, chickpea, grasspea, and pasture and forage crops are distributed by the International Center for Agricultural Research in the Dry Areas (ICARDA) every year to around 70 countries. New accessions are added to the germplasm collections in the Center's genebank and utilized in the breeding programs. Recent years have witnessed an increasing global concern about the loss of plant genetic resources because of conflicts, human pandemic diseases, extreme weather events, pest and disease outbreaks, and natural calamities such as earthquakes, floods, etc., which led to disrupting access to germplasm and undermining social protection systems. Safety duplication of germplasm collections held in genebanks in other institutions, including international, regional, and national genebanks, as well as the Svalbard Global Seed Vault (SGSV), is one of the essential measures to safeguard germplasm and also to replenish any lost accessions and resume use and distribution of seeds to users internationally. Germplasm distribution procedures are conducted per International Plant Protection Convention phytosanitary regulations to avoid the transboundary spread of seed-borne pests and pathogens. The ICARDA’s Seed Health Laboratory exercises maximum efforts to maintain the “phytosanitary clean” health status of germplasm during regeneration, conservation, distribution and ensure compliance with phytosanitary regulations in international germplasm distributions to guarantee minimum loss of genetic resources. These efforts include the development of new methods to detect and manage seed-borne pathogens. An increase in global awareness to preserve germplasm for current and future use is crucial to combat climate challenge, malnutrition, and food insecurity.

Molecular and Pathological Characterization of Classical Swine Fever Virus Genotype 2 Strains Responsible for the 2013-2018 Outbreak in Colombia.

Classical swine fever (CSF) is a highly contagious transboundary viral disease of domestic and wild pigs. Despite mass vaccination and continuous eradication programs, CSF remains endemic in Asia, some countries in Europe, the Caribbean and South America. Since June 2013, Northern Colombia has reported 137 CSF outbreaks, mostly in backyard production systems with low vaccination coverage. The purpose of this study was to characterize the virus responsible for the outbreak. Phylogenetic analysis based on the full-length E2 sequence shows that the virus is closely related to CSF virus (CSFV) genotype 2.6 strains circulating in Southeast Asia. The pathotyping experiment suggests that the virus responsible is a moderately virulent strain. The 190 nucleotide stretch of the E2 hypervariable region of these isolates also shows high similarity to the CSFV isolates from Colombia in 2005 and 2006, suggesting a common origin for the CSF outbreaks caused by genotype 2.6 strains. The emergence of genotype 2.6 in Colombia suggests a potential transboundary spread of CSFV from Asia to the Americas, complicating the ongoing CSF eradication efforts in the Americas, and emphasizes the need for continuous surveillance in the region.

Whole-genome sequencing for One Health surveillance of antimicrobial resistance in conflict zones: a case study of Salmonella spp. and Campylobacter spp. in the West Bank, Palestine.

Antimicrobial resistance (AMR) is a critical global concern driven by the overuse, misuse, and/or usage of inadequate antibiotics on humans, animals' agriculture, and as a result of contaminated environments. This study is the first One Health survey in the Middle East that incorporated whole-genome sequencing (WGS) to examine the spread of AMR in Campylobacter spp. and Salmonella spp. This cross-sectional study was conducted to examine the role of AMR at the human-animal-environmental interface and was performed in Ramallah/Al-Bireh and Jerusalem governorates of the central West Bank, Palestine. In 2021 and 2022, a total of 592 samples were collected and analyzed. From a total of 65 Campylobacter jejuni and 19 Salmonella spp. isolates, DNA was extracted for WGS using Oxford Nanopore Technologies MinION platform. We found that the dominant serotypes of C. jejuni and Salmonella enterica were present in chicken manure, chicken meat sold in markets, and feces of asymptomatic farm workers, with high genetic similarities between the isolates regardless of origin. Additionally, our results showed rapid strain turnover in C. jejuni from the same sites between 2021 and 2022. Most of the positive Salmonella spp. samples were multidrug-resistant (MDR) S. enterica serovar Muenchen carrying the plasmid of emerging S. infantis (pESI) megaplasmid, conferring resistance to multiple antibiotics. Our findings highlight the spread of MDR foodborne pathogens from animals to humans through the food chain, emphasizing the importance of a One Health approach that considers the interconnections between human, animal, and environmental health. IMPORTANCE Prior to this study, there existed hardly an integrated human-animal-environmental study of Salmonellosis and Campylobacteriosis and related AMR in Middle Eastern countries. The few existing studies lack robust epidemiological study designs, adequate for a One Health approach, and did not use WGS to determine the circulating serotypes and their AMR profiles. Civil unrest and war in Middle Eastern countries drive AMR because of the breakdown of public health and food security services. This study samples simultaneously humans, animals, and the environment to comprehensively investigate foodborne pathogens in the broiler chicken production chain in Palestine using WGS. We show that identical serotypes of C. jejuni and S. enterica can be found in samples from chicken farms, chicken meat sold in markets, and asymptomatic broiler chicken production workers. The most striking feature is the rapid dynamic of change in the genetic profile of the detected species in the same sampling locations. The majority of positive Salmonella spp. samples are MDR S. enterica serovar Muenchen isolates carrying the pESI megaplasmid. The results demonstrate a close relationship between the S. enterica serovar Muenchen isolates found in our sample collection and those responsible for 40% of all clinical Salmonella spp. isolates in Israel as previously reported, with a sequence identity of over 99.9%. These findings suggest the transboundary spread of MDR S. enterica serovar Muenchen strains from animals to humans through the food chain. The study underscores the importance of combining integrated One Health studies with WGS for detecting environmental-animal-human transmission of foodborne pathogens that could not be detected otherwise. This study showcases the benefits of integrated environmental-animal-human sampling and WGS for monitoring AMR. Environmental samples, which may be more accessible in conflict-torn places where monitoring systems are limited and regulations are weak, can provide an effective AMR surveillance solution. WGS of bacterial isolates provides causal inference of the distribution and spread of bacterial serotypes and AMR in complex social-ecological systems. Consequently, our results point toward the expected benefits of operationalizing a One Health approach through closer cooperation of public and animal health and food safety authorities.

Home ranges and movement patterns of wild boars (Sus scrofa) at the northern edge of the species’ distribution range

In Finland, the wild boar (Sus scrofa L.) lives on the northernmost edge of the species’ distribution range, and the population mainly originates from individuals immigrating from Russia. Most of the current population lives in the southeastern part of the country, and the wild boar is steadily expanding further. To develop effective risk and population management in novel northern wild boar regions, detailed information about the species’ local ecology is required. To estimate wild boar movement patterns, 17 adult wild boars were monitored using GPS collars from May 2020 to September 2022 in the core region of the current distribution. The average total home ranges of wild boars (87.1 ± 17 km2 MCP, 33 ± 5.5 SE km2 95% KDE) were larger compared with studies from southern latitudes. The length of nocturnal activity times varied seasonally. All studied individuals at the border zone (N = 15) showed continuous transboundary movements, with home range core areas located mainly on the Russian side. Wild boar locations were predominantly in the Russia border zone, especially resting sites during the daytime. Most locations in Finland were from night-time feeding excursions. Our study shows that, although home ranges are large, adult wild boars are relatively sedentary also in northern latitudes. However, the movement capacity of the species enables the transboundary spread of diseases such as African swine fever. Our results provide information for risk management and emphasize the importance of transboundary collaboration in the monitoring and management of common wild boar populations.

Host range, severity and trans boundary transmission of Orf virus (ORFV).

Contagious ecthyma in small ruminants is a zoonotic disease caused by Orf virus (ORFV) in the genus Parapoxvirus that can be deadly to its natural hosts. It causes significant losses worldwide, and commonly infects humans. However, the literature about its comparative severity in sheep and goat hosts is misleading; and while contagious ecthyma has been shown to occur in camels and transmit to humans, there is confusion as to whether ORFV is responsible. Camels are important from a 'One Health' perspective as they have been implicated as a reservoir host for the virus causing Middle East Respiratory Syndrome (MERS), which has a case fatality rate of 35% in humans. We compared ORFV gene sequences and mortality data from the West Bank in Palestine, where ORFV has not been reported previously, with data from the region. Surprisingly, we found that infections of camels that had been attributed to ORFV were more closely related to a different member of the genus Parapoxvirus. Two Middle East ORFVs isolated from humans were unrelated and sat alongside sheep and goat derived sequences on two distinct ORFV lineages of a maximum likelihood B2L gene tree. One of the viral lineages bifurcated to produce a monophyletic group of goat-derived ORFVs characterized uniquely by a glycine at amino acid reside 249. We found that serine is the ancestral allele shared between ORFV infections of sheep and also two closely related Parapoxviruses (PCPV and CCEV), indicating that the glycine allele represents a more recent shift in virus host range adaptation to goats. Furthermore, and contrary to some reports that ORFV is more severe in goats than in sheep, we observed median mortality of up to 24.5% in sheep, but none in goats. We also identified trans-boundary spread of ORFV between the West Bank and Israel.

The Spread of Peste Des Petits Ruminants Virus Lineage IV in West Africa.

Monitoring the transboundary spread of peste des petits ruminants (PPR) virus is an essential part of the global efforts towards the eradication of PPR by 2030. There is growing evidence that Lineage IV is becoming the predominant viral lineage, replacing Lineage I and II in West Africa. As part of a regional investigation, samples collected in Burkina Faso, Côte d'Ivoire, Guinea and Ghana were screened for the presence of PPRV. A segment of the nucleoprotein gene from positive samples was sequenced, and phylogenetic analysis revealed the co-circulation of Lineage II and IV in Burkina Faso, Côte d'Ivoire and Guinea, and the identification of Lineage IV in Ghana. These data will be of importance to local and regional authorities involved in the management of PPRV spread.

Partial validation of a TaqMan quantitative polymerase chain reaction for the detection of the three genotypes of Infectious spleen and kidney necrosis virus.

Megalocytiviruses (MCVs) are double-stranded DNA viruses known to infect important freshwater and marine fish species in the aquaculture, food, and ornamental fish industries worldwide. Infectious spleen and kidney necrosis virus (ISKNV) is the type species within the genus Megalocytivirus that causes red seabream iridoviral disease (RSIVD) which is a reportable disease to the World Animal Health Organization (WOAH). To better control the transboundary spread of this virus and support WOAH reporting requirements, we developed and partially validated a TaqMan real-time qPCR assay (ISKNV104R) to detect all three genotypes of ISKNV, including the two genotypes that cause RSIVD. Parameters averaged across 48 experiments used a 10-fold dilution series of linearized plasmid DNA (107-101 copies), carrying a fragment of the three-spot gourami iridovirus (TSGIV) hypothetical protein revealed that the assay was linear over 7 orders of magnitude (107-101), a mean efficiency of 99.97 ± 2.92%, a mean correlation coefficient of 1.000 ± 0.001, and a limit of detection (analytical sensitivity) of ≤10 copies of TSGIV DNA. The diagnostic sensitivity and specificity for the ISKNV104R qPCR assay was evaluated and compared to other published assays using a panel of 397 samples from 21 source populations with different prevalence of ISKNV infection (0-100%). The diagnostic sensitivity and specificity for the ISKNV104R qPCR assay was 91.99% (87.28-95.6; 95% CI) and 89.8% (83.53-94.84). The latent class analysis showed that the ISKNV104R qPCR assay had similar diagnostic sensitivities and specificities with overlapping confidence limits compared to a second TaqMan qPCR assay and a SYBR green assay. This newly developed TaqMan assay represents a partially validated qPCR assay for the detection of the three genotypes of the species ISKNV. The ISKNV104R qPCR assay once fully validated, will serve as an improved diagnostic tool that can be used for ISKNV surveillance efforts and diagnosis in subclinical fish to prevent further spread of MCVs throughout the aquaculture and ornamental fish industries.

Presence of Anaplasma phagocytophilum Ecotype I in UK Ruminants and Associated Zoonotic Risk.

Anaplasma phagocytophilum is the causative agent of tick-borne fever in sheep, pasture fever in cattle, and granulocytic anaplasmosis in humans. The increasing prevalence and transboundary spread of A. phagocytophilum in livestock, ticks, and wildlife in the UK poses a potential zoonotic risk that has yet to be estimated. Several ecotypes of A. phagocytophilum show variable zoonotic potential. To evaluate the possible risk associated with the transmission of A. phagocytophilum from ruminants to humans, the ecotype was determined by sequencing the groEL gene from 71 positive blood and tissue samples from UK ruminants. Thirty-four groEL sequences were obtained, fourteen of which were identified in multiple samples. Of the 13 nucleotide polymorphisms identified through pairwise comparison, all corresponded to synonymous substitutions. The subsequent phylogenetic estimation of the relationship with other European/world isolates indicated that all the groEL sequences clustered with other ecotype I sequences. The presence of ecotype I closely reflects that observed in ruminants in continental Europe and suggests a lower risk of zoonotic transmission from this reservoir.

Leveraging livestock movements to urban slaughterhouses for wide-spread Rift Valley fever virus surveillance in Western Kenya

Rift Valley fever virus (RVFV) is an economically devastating, zoonotic arbovirus endemic across Africa with potential to cause severe disease in livestock and humans. Viral spread is primarily driven by movement of domestic ruminants and there is a high potential for transboundary spread. Despite influx of livestock to urban areas in response to the high demand for meat and animal products, RVFV has not been detected in any urban center. The objectives of this study were to determine the feasibility of assessing risk of RVFV introduction to urban Kisumu, Kenya, by testing slaughtered livestock for RVFV exposure and mapping livestock origins. Blood was collected from cattle, sheep, and goats directly after slaughter and tested for anti-RVFV IgG antibodies. Slaughterhouse businessmen responded to a questionnaire on their individual animals' origin, marketplace, and transport means. Thereafter, we mapped livestock flow from origin to slaughterhouse using participatory methods in focus group discussions with stakeholders. Qualitative data on route choice and deviations were spatially integrated into the map. A total of 304 blood samples were collected from slaughtered livestock in October and November 2021. Most (99%) of animals were purchased from 28 different markets across eight counties in Western Kenya. The overall RVFV seroprevalence was 9% (19% cattle, 3% in sheep, and 7% in goats). Migori County bordering Tanzania had the highest county-level seroprevalence (34%) and 80% of all seropositive cattle were purchased at the Suba Kuria market in Migori County. Road quality and animal health influenced stakeholders' decisions for choice of transport means. Overall, this proof-of-concept study offers a sampling framework for RVFV that can be locally implemented and rapidly deployed in response to regional risk. This system can be used in conjunction with participatory maps to improve active livestock surveillance and monitoring of RVFV in Western Kenya, and these methods could be extrapolated to other urban centers or livestock diseases.

Characterization and epidemiological analysis of Vibrio parahaemolyticus isolated from different marine products in East China

Vibrio parahaemolyticus is a major foodborne pathogen with a wide distribution in the world that causes economic and public health problems. Here, we isolated 152 V. parahaemolyticus strains from shellfish, shrimp, crab, and snails from 5 provinces in East China, and analyzed the genetic diversity, population structure, and virulence profiles of these isolates. Our results showed that the 152 V. parahaemolyticus strains belong to 84 different sequence types (STs), of which 69 (82.14 %) STs and 60 alleles were newly identified. The thermostable direct hemolysin (TDH) and the TDH-related hemolysin (TRH) were present in 4 V. parahaemolyticus isolates (2.63 %) respectively, while toxRS/new, a distinctive toxRS sequence that was associated with pandemic V. parahaemolyticus strains, is present in 52 isolates (34.21 %). Thereinto, both the ZJ11 and ZJ12 strains measure up to the standard of toxRS/new+, tdh+, and trh-, which was widely used marker for the rapid screening of pandemic strains, and thus these strains may have the risk of infectious outbreaks. In addition, we observed that all the 152 V. parahaemolyticus isolates encode type III secretion systems 1 and type VI secretion system (T6SS) 2, while 119 isolates (78.29 %) of them also contain T6SS1. The genetic relatedness of our isolates to the human V. parahaemolyticus collection was explored, which showed that ST6, ST79, ST162, ST1060, and ST1061 were all identified in both human isolates (7 isolates) uploaded in the PubMLST database and our marine products isolates (7 isolates). Our findings expand the views of the genetic diversity of V. parahaemolyticus and will contribute to understanding the potential risk of the transboundary spread of this bacterium.

Role of wild birds in the spread of clade 2.3.4.4e H5N6 highly pathogenic avian influenza virus into South Korea and Japan.

H5Nx highly pathogenic avian influenza viruses (HPAIVs) have caused transboundary epizootics in poultry and wild birds. In 2016, the H5N6 subtype of clade 2.3.4.4e HPAIVs caused multiple outbreaks in Asia, including China, Japan, Korea, and Vietnam. However, the geographical spread pattern of 2.3.4.4e H5N6 HPAIV has not been clearly identified. To better understand the emergence and transmission history of 2.3.4.4e H5N6 HPAIV, we investigated the underlying epidemiologic processes associated with this viral spread by performing a Bayesian phylogeography analysis. The results revealed that wild waterfowl played a central role in the transboundary spread of clade 2.3.4.4e H5N6 HPAIV into both endemic and non-endemic countries, causing multiple incursions of the 2.3.4.4e H5N6 HPAIV into South Korea, Japan, and Vietnam. In our analysis, Guangdong province, China was estimated to be the most probable site where 2.3.4.4e H5N6 HPAIVs emerged prior to the transboundary transmissions. Continued genomic surveillance in both wild birds and poultry would be necessary for monitoring of HPAIV incursions. In addition, enhanced biosecurity would be key to preventing the HPAIV spread by minimizing contact between domestic poultry and wild birds.

A highly adaptable platform powered by CRISPR-Cas12a to diagnose lumpy skin disease in cattle

Lumpy skin disease (LSD) in cattle, a transboundary viral disease of cattle once restricted to Africa, has been spreading to many European and Asian countries in the past decade with huge economic losses. This emerging worldwide threat to cattle warrants the development of diagnostic methods for accurate disease screening of suspected samples to effectively control the spread of LSD. In this study, we integrated pre-amplification and three kinds of sensor systems with CRISPR and therefore established an LSD diagnosis platform with highly adaptable and ultra-sensitive advantages. It was the first CRISPR-powered platform that could identify lumpy skin disease virus from vaccine strains of goat pox virus and sheep pox virus. Its limit of detection (LOD) was one copy/reaction after introducing PCR or recombinase-aided amplification (RAA). Moreover, this platform achieved a satisfactory overall agreement in clinical diagnoses of 50 samples and its reproducibility and accuracy were superior to other qPCR methods we tested. The whole diagnostic procedure, from DNA extraction to the results, could complete in 5 h with a total cost of 1.7–9.6 $/test. Overall, this CRISPR-powered platform provided a novel diagnostic tool for portable, ultra-sensitive, rapid, and highly adaptable disease screening of LSD and may be an effective method to control this transboundary disease's spread.

Seroprevalence of Enzootic Teschen Disease in the Wild Boar Population in Ukraine.

Teschen disease is an acute fatal enterovirus encephalomyelitis of pigs, characterized by a range of central nervous system disorders. The cause of porcine enterovirus encephalomyelitis is the picornavirus porcine teschovirus-1 (PTV-1). There are at least 12 disctinct serotypes of PTVs, where PTV-2 to PTV-12 serogroups are associated with other forms of disease (Talfan disease or poliomyelitis suum) or benign enzootic paresis. Combined, PTVs have been found to have a high seroprevalence, up to 65%, in healthy pig populations in Europe. PTVs have also been detected in wild boar, including the divergent PTV-13 serogroup; wild suids may represent a sylvatic reservoir capable of carrying the virus long distances. In Ukraine, Teschen disease is widespread and causes lethal disease in domestic pigs. To understand temporal and geographical distribution of Teschen disease virus (PTV-1) in wild boar in Ukraine (2001-2013), we analyzed seroprevalence of 6840 blood serum samples from hunted suids using a virus microneutralization assay. A total of 1364 samples (19.9%) were seropositive, with average antibody titer ratios 5.89 ± 0.03 log2 (range 5-12 log2). Teschen seroprevalence was temporally and geographically concentrated in the northern and western regions of Ukraine, corresponding to forested regions (polissya) and overlapping with wild boar populations and habitats, suggesting endemicity in wild boar. The virus sporadically emerged in central, southern, and eastern forested regions, suggesting long-distance movement of infected wild suids. Thus, wild boar should be monitored for potential transboundary spread in forested and mountain regions and spillover of PTVs to domestic swine populations.

Azole-resistant Aspergillus fumigatus as an emerging worldwide pathogen.

Aspergillus fumigatus, a ubiquitous pathogen, causes aspergillosis in humans, especially in immunodeficient patients. Azoles are frontline antifungal drugs for treating aspergillosis. The recent global emergence of azole resistance in A. fumigatus has become a serious problem worldwide. It has arisen through two routes: long-term azole medical therapy, called the patient route, and the use of azole fungicides in its habitats especially for agricultural activities, called the environmental route. Resistant strains developed through the latter route show cross-resistance to medical azoles because of the identical molecular target Cyp51A between azole compounds used for medical treatment and agricultural disease control. In azole-resistant strains arising through the environmental route, A. fumigatus is observed frequently to possess mutations in the cyp51A gene linked to tandem repeats in the promoter region such as TR34 /L98H and TR46 /Y121F/T289A. The results of microsatellite genotyping analyses of resistant A. fumigatus strains have suggested a transboundary spread of this microorganism in many countries. Diverse actors are involved in the global highway of transmission. Therefore, the matter must be addressed as a "One Health" issue. This review presents a background of azole resistance in A. fumigatus and introduces newly discovered difficulties generated as this pathogen spreads worldwide.

Cover Image

Cover caption: The cover image is based on the Editorial Feed: A new pathway for the domestic and transboundary spread of viral pathogens of veterinary significance by Scott Dee and Gordon Spronk, https://doi.org/10.1111/tbed.14349.