Wide Host Range Research Articles

Bacillus integrative plasmid system combining a synthetic gene circuit for efficient genetic modifications of undomesticated Bacillus strains

BackgroundOwing to CRISPR-Cas9 and derivative technologies, genetic studies on microorganisms have dramatically increased. However, the CRISPR-Cas9 system is still difficult to utilize in many wild-type Bacillus strains owing to Cas9 toxicity. Moreover, less toxic systems, such as cytosine base editors, generate unwanted off-target mutations that can interfere with the genetic studies of wild-type strains. Therefore, a convenient alternative system is required for genetic studies and genome engineering of wild-type Bacillus strains. Because wild-type Bacillus strains have poor transformation efficiencies, the new system should be based on broad-host-range plasmid-delivery systems.ResultsHere, we developed a Bacillus integrative plasmid system in which plasmids without the replication initiator protein gene (rep) of Bacillus are replicated in a donor Bacillus strain by Rep proteins provided in trans but not in Bacillus recipients. The plasmids were transferred to recipients through a modified integrative and conjugative element, which is a wide host range plasmid-delivery system. Genetic mutations were generated in recipients through homologous recombination between the transferred plasmid and the genome. The system was improved by adding a synthetic gene circuit for efficient screening of the desired mutations by double crossover recombination in recipient strains. The improved system exhibited a mutation efficiency of the target gene of approximately 100% in the tested wild-type Bacillus strains.ConclusionThe Bacillus integrative plasmid system developed in this study can generate target mutations with high efficiency when combined with a synthetic gene circuit in wild-type Bacillus strains. The system is free of toxicity and unwanted off-target mutations as it generates the desired mutations by traditional double crossover recombination. Therefore, our system could be a powerful tool for genetic studies and genome editing of Cas9-sensitive wild-type Bacillus strains.

Genome Sequence Resource of Fusarium graminearum TaB10 and Fusarium avenaceum KA13, Causal Agents of Stored Apple Rot.

The filamentous fungus Fusarium graminearum is a well-known cereal pathogen and F. avenaceum is a pathogen with a wide host range. Recently, both species were reported as causal agents of apple rot, raising concerns about postharvest yield losses and mycotoxin contamination. Here, we report genome assemblies of F. avenaceum KA13 and F. graminearum TaB10, both isolated from fruits with symptoms of apple rot. The final F. avenaceum KA13 genome sequence assembly of 41.7 Mb consists of 34 scaffolds, with an N50 value of 2.2 Mb and 15,886 predicted genes. The total size of the final F. graminearum TaB10 assembly is 36.76 Mb, consisting of 54 scaffolds with an N50 value of 1.7 Mb, and it consists of 14,132 predicted genes. These new genomes provide valuable resources to better understand plant-microbe interaction in stored apple rot disease. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Revisiting fecal metatranscriptomics analyses of macaques with idiopathic chronic diarrhoea with a focus on trichomonad parasites.

Trichomonads, anaerobic microbial eukaryotes members of the phylum Parabasalia, are common obligate extracellular symbionts that can lead to pathological or asymptomatic colonization of various mucosal surfaces in a wide range of animal hosts. Results from previous in vitro studies have suggested a number of intriguing mucosal colonization strategies by Trichomonads, notably highlighting the importance of interactions with bacteria. However, in vivo validation is currently lacking. A previous metatranscriptomics study into the cause of idiopathic chronic diarrhoea in macaques reported the presence of an unidentified protozoan parasite related to Trichomonas vaginalis. In this work, we performed a reanalysis of the published data in order to identify the parasite species present in the macaque gut. We also leveraged the information-rich metatranscriptomics data to investigate the parasite behaviour in vivo. Our results indicated the presence of at least 3 genera of Trichomonad parasite; Tetratrichomonas, Pentatrichomonas and Trichomitus, 2 of which had not been previously reported in the macaque gut. In addition, we identified common in vivo expression profiles shared amongst the Trichomonads. In agreement with previous findings for other Trichomonads, our results highlighted a relationship between Trichomonads and mucosal bacterial diversity which could be influential in health and disease.

PLATYNOTA STULTANA WALSINGHAM, 1884 (LEPIDOPTERA TORTRICIDAE) FOUND IN ITALY, INVASIVE PEST IN EUROPE

Notes on the presence of the Omnivorous Leafroller, Platynota stultana (Lepidoptera Tortricidae), in Italy and its presence and interception in other European countries are reported. Geographical distributions and principal morphological and biological characters are illustrated. Considering the wide host range of P. stultana, this pest has the potential to damage many cultivated and wild plants in the European and Mediterranean Plant Protection Organization (EPPO) region. Its interception in England, the Netherlands, and Germany and its introduction into Spain and Italy clearly demonstrates that it has the potential to enter and establish itself in the Southern and Mediterranean parts of Europe. Key Words: Platynota stultana, Lepidoptera Tortricidae, Italy, Europe, pest risk.

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Staphylococcus virus ΦSA012 has a wide host range and efficient lytic activity. Here, we assessed the biological stability of ΦSA012 against temperature, freeze-thawing, and pH to clinically apply the phage. In addition, inoculation of ΦSA012 through i.p. and i.v. injections into mice revealed that phages were reached the limit of detection in serum and accumulated notably spleens without inflammation at 48h post-inoculation. Furthermore, inoculation of ΦSA012 through s.c. injections in mice significantly induced IgG, which possesses neutralizing activity against ΦSA012 and other Staphylococcus viruses, ΦSA039 and ΦMR003, but not Pseudomonas viruses ΦS12-3 and ΦR18 or Escherichia viruses T1, T4, and T7 in vitro. Immunoelectron microscopic analysis showed that purified anti-phage IgG recognizes the long-tail fiber of staphylococcus viruses. Although S. aureus inoculation resulted in a 25% survival rate in a mouse i.p. model, ΦSA012 inoculation (i.p.) improved the survival rate to 75%; however, the survival rate of ΦSA012-immunized mice decreased to less than non-immunized mice with phage i.v. injection at a MOI of 100. These results indicated that ΦSA012 possesses promise for use against staphylococcal infections but we should carefully address the appropriate dose and periods of phage administration. Our findings facilitate understandings of staphylococcus viruses for phage therapy.

Serendipita indica-A Review from Agricultural Point of View.

Fulfilling the food demand of a fast-growing population is a global concern, resulting in increased dependence of the agricultural sector on various chemical formulations for enhancing crop production. This leads to an overuse of chemicals, which is not only harmful to human and animal health, but also to the environment and the global economy. Environmental safety and sustainable production are major responsibilities of the agricultural sector, which is inherently linked to the conservation of the biodiversity, the economy, and human and animal health. Scientists, therefore, across the globe are seeking to develop eco-friendly and cost-effective strategies to mitigate these issues by putting more emphasis on the use of beneficial microorganisms. Here, we review the literature on Serendipita indica, a beneficial endophytic fungus, to bring to the fore its properties of cultivation, the ability to enhance plant growth, improve the quality of produced crops, mitigate various plant stresses, as well as protect the environment. The major points in this review are as follows: (1) Although various plant growth promoting microorganisms are available, the distinguishing character of S. indica being axenically cultivable with a wide range of hosts makes it more interesting for research. (2) S. indica has numerous functions, ranging from promoting plant growth and quality to alleviating abiotic and biotic stresses, suggesting the use of this fungus as a biofertiliser. It also improves the soil quality by limiting the movement of heavy metals in the soil, thus, protecting the environment. (3) S. indica's modes of action are due to interactions with phytohormones, metabolites, photosynthates, and gene regulation, in addition to enhancing nutrient and water absorption. (4) Combined application of S. indica and nanoparticles showed synergistic promotion in crop growth, but the beneficial effects of these interactions require further investigation. This review concluded that S. indica has a great potential to be used as a plant growth promoter or biofertiliser, ensuring sustainable crop production and a healthy environment.

Current control options and a way towards risk-based control of Toxoplasma gondii in the meat chain

Toxoplasma gondii is a zoonotic protozoan with a wide range of hosts and cosmopolitan distribution. Due to its impact on human and animal health, as well as related economic losses, T. gondii is considered as one of the most significant foodborne pathogens nowadays. Consumption of raw or undercooked meat is a well known risk factor for human infection. However, there is no legislation regarding control of T. gondii in the meat chain, and current meat safety assurance systems (MSAS) have little control over this parasite. Traditional meat inspection is not capable of detecting microscopic cysts of T. gondii in the tissues of slaughter animals, and available laboratory tests are still not sufficiently sensitive or specific to detect infection in individual carcasses. Possible solutions for overcoming these challenges and gaining a higher level of control over T. gondii in the meat chain come in the form of a risk-based meat safety assurance system (RB-MSAS) that incorporates risk-categorisation of farms and targeted interventions, as well as education of all interested parties.

Mycoplasma ovipneumoniae: A Most Variable Pathogen.

Mycoplasma ovipneumoniae, a well-established respiratory pathogen of sheep and goats, has gained increased importance recently because of its detection in wild ruminants including members of the Cervidae family. Despite its frequent isolation from apparently healthy animals, it is responsible for outbreaks of severe respiratory disease which are often linked to infections with multiple heterologous strains. Furthermore, M. ovipneumoniae is characterized by an unusually wide host range, a high degree of phenotypic, biochemical, and genomic heterogeneity, and variable and limited growth in mycoplasma media. A number of mechanisms have been proposed for its pathogenicity, including the production of hydrogen peroxide, reactive oxygen species production, and toxins. It shows wide metabolic activity in vitro, being able to utilize substrates such as glucose, pyruvate, and isopropanol; these patterns can be used to differentiate strains. Treatment of infections in the field is complicated by large variations in the susceptibility of strains to antimicrobials, with many showing high minimum inhibitory concentrations. The lack of commercially available vaccines is probably due to the high cost of developing vaccines for diseases in small ruminants not presently seen as high priority. Multiple strains found in affected sheep and goats may also hamper the development of effective vaccines. This review summarizes the current knowledge and identifies gaps in research on M. ovipneumoniae, including its epidemiology in sheep and goats, pathology and clinical presentation, infection in wild ruminants, virulence factors, metabolism, comparative genomics, genotypic variability, phenotypic variability, evolutionary mechanisms, isolation and culture, detection and identification, antimicrobial susceptibility, variations in antimicrobial susceptibility profiles, vaccines, and control.

Direct and Endophytic Effects of Fungal Entomopathogens for Sustainable Aphid Control: A Review

Entomopathogenic fungi (EPF) are cosmopolitan species of great interest in pest management due to their ability to cause epizooty in soil-dwelling and aboveground insects. Besides their direct effect against a wide host range of serious agricultural insect pests, such as aphids, a major emphasis has been placed on investigating the impact of EPF with endophytic traits (EIPF) on aboveground tripartite interactions between host plants, herbivores and beneficial insects. However, despite their valuable role in biocontrol processes, there is still more to explore about their diverse potential as ecofriendly biological control agents. Herein, we provide an overview of the meaningful role and faced challenges following the use of EPF and EIPF to control aphids.

Identification of a Multidrug Resistant Pseudomonas aeruginosa Isolate Harboring Infrequent Red Fluorescence Plasmid from COPD Patient.

Pseudomonas aeruginosa is a notorious Gram-negative opportunistic pathogen that normally causes acute and chronic infections in a wide range of hosts. In this study, a multi-resistant P. aeruginosa isolate L1a harboring an infrequent plasmid with red fluorescence was obtained from the bronchoalveolar lavage fluid of a patient with chronic obstructive pulmonary disease. The results of susceptibility testing and virulence-related phenotypic identification revealed that P. aeruginosa L1a was resistant to levofloxacin, cefepime, aztreonam, and imipenem and showed significantly stronger capacities for swimming and pyocyanin production than the reference strain PAO1. The genome of P. aeruginosa L1a was assembled into one circular chromosome (6,216,913 bp) and one circular plasmid (9111 bp). P. aeruginosa L1a was found to belong to the multilocus sequence type ST549, and serotype O5, and carried 8 drug resistance genes and 18 multidrug efflux pump-related genes in the chromosomal DNA. The plasmid pL1a harbored a tetracycline resistant gene tetA and a functional red fluorescence protein. This study reports a multidrug resistant P. aeruginosa clinical isolate harboring an infrequent red fluorescence plasmid for the first time.

Serological detection of the latent infection of Brucellosis in calves in Mosul city, Iraq

Brucellosis is a widespread and endemic disease in the Middle East and Mediterranean regions, and it has significant public health and livestock industry concerns. It can affect a wide range of mammalian hosts, including humans. This study aimed to determine the seroprevalence of thelatent infection of brucellosisin calves; Rose Bengal and indirect ELISA tests were conducted for this purpose. The serum samples were collected from 184 local calves of both sexes aged between 1 to 8 months (92 males and 92 females) from different regions of Mosul city. The current study revealed that all samples tested negative with the Rose Bengal test, whereas, in indirect ELISA 5.9%, the samples were against Brucella species. The seroprevalence of Brucella antibodies showed no statistical differences between males and females, which was 5.4 and 6.5%, respectively. This study concluded that the indirect ELISA test showed high diagnostic efficacy in detecting anti-Brucella antibodies in young calves.

PENGARUH CAIRAN PERASAN BROTOWALI (Tinospora cripsa L.) TERHADAP MORTALITAS ULAT GRAYAK (Spodoptera litura L.) DI LABORATORIUM

Armyworm (Spodoptera litura L.) is an insect pest that is difficult to control couse its rapid reproduction and has a wide host range. One of alternative to control by using vegetable pesticides which is brotowali stems (Tinospora crispa L.). This study used a completely randomized design (CRD) consisting of 5 treatments and 5 replications. The treatments were 8 ml, 12 ml, 16 ml, 20 and 40 ml. The observation variable was mortality percentage of S. litura. The results showed that brotowali juice was treated with 40 mL/L water (A4) with a mortality percentage of S. litura 63.67% at 24 hours after application. Brotowali Juice has effect to Spodoptera litura mortality by mortality percentage 63,67%.

Establishment and spread of the invasive mite, Tetranychus gloveri Banks (Prostigmata: Tetranychidae) in Kerala, India

Tetranychus gloveri Banks was first recorded as Tetranychus okinawanus Ehara in India on an ornamental plant, Adenium obesum from Thrissur district, Kerala state, during 2016 (Zeity et al. 2016). Recently, Sharkey et al. (2022) synonymized T. okinawanus with T. gloveri based on morphological and molecular data. In Kerala, T. gloveri has emerged as a predominant species of mite infesting major agricultural and horticultural crops in the district (Arunima et al. 2018). In order to investigate the distribution and host range of the mite species in Kerala, periodical surveys were conducted in different agricultural ecosystems across the state during March 2020 to June 2022. Spider mite infested samples were collected from fruit crops, vegetables, ornamental plants, medicinal plants and other non-crop plants from different localities, kept in polythene bags. The GPS data of the locality and host plants were recorded. In the laboratory, a single gravid female from each sample was used to establish isoline culture providing unique accession number. Male and female specimens from each isoline culture were slide mounted on Hoyer’s medium and morphological characterization of the slide mounted mite specimens were carried to establish the species identity. Characters such as chaetotaxy of hysterosoma and legs, structure of empodium and pattern of dorsal striae between e1 and f1 of female were used for genus level identification, while the shape of male genitalia, aedeagus, was used for species level identification. In this study, T. gloveri was recorded from a wide host range of 35 host plants in 24 plant families viz., Malvaceae, Cucurbitaceae, Fabaceae, Amaranthaceae, Rutaceae, Solanaceae, Musaceae, Moraceae, Anacardiaceae, Caricaceae, Adoxaceae, Rosaceae, Compositae, Gentianaceae, Convolvulaceae, Balsaminaceae, Orchidaceae, Asparagaceae, Goodeniaceae, Apocyanaceae, Euphorbiaceae, Oxalidaceae, Lamiaceae and Pontederiaceae. The mite species was distributed in seven districts covering northern, central and southern regions viz., Wayanad, Malappuram, Thrissur, Palakkad, Ernakulam, Alappuzha and Thiruvananthapuram districts of Kerala. Curry leaf, country kreat (Exacum bicolor), sunflower, Victoria corn plant (Dracaena sp.), Calotropis gigantea, little tree plant (Biophytum sensitivum), holy basil (Ocimum sanctum) and the aquatic pickerel weed (Monochoria vaginalis) are new host records for T. gloveri. The study confirms that the mite species has established and spread across Kerala by widening its host range and expanding its geographical area of distribution, within a short span after its introduction into the state.

Investigating the survival of herpes simplex virus on toothbrushes and surrogate phallic devices.

Herpes simplex virus-1 (HSV-1) is a member of the alphaherpesvirus (alphaherpesvirinae) subfamily, allowing it to affect a wide range of hosts. Herpes simplex virus-1 affects 3.7 billion people, or 67% of the population, under the age of 50. With a vast number of people infected by the virus, everyday objects are often contaminated with this agent. In this study we determined how long HSV-1 can remain viable on contaminated fomites. Fomites were selected for their use near potentially contaminated orifices and variable frequency of sanitization. Toothbrushes and surrogate phallic devices (SPDs) were cut, sterilized, and contaminated. After contaminating the fomites, we collected samples over a 24h period, then used plaque assays to determine viral titers at prescribed time points. The quantity of replication-competent virus present appears to decrease significantly 2h post-contamination, then steadily declines over time, nearing zero at 24h. Our findings suggest that different surfaces influence HSV-1 survival. Proper cleaning must be performed for these types of fomites, especially if shared in an environment where someone with active genital or oral herpes lesions uses one of these fomites shortly after someone else.

Infectious droplet exposure is an inefficient route for SARS-CoV-2 infection in the ferret model.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19) in humans, has a wide host range, naturally infecting felids, canids, cervids, rodents and mustelids. Transmission of SARS-CoV-2 is universally accepted to occur via contact with contaminated secretions from the respiratory epithelium, either directly or indirectly. Transmission via droplet nuclei, generated from a cough or sneeze, has also been reported in several human and experimental animal scenarios. However, the role of droplet transmission at the human-animal interface remains to be fully elucidated. Here, the ferret infection model was used to investigate the routes of infection for the SARS-CoV-2 beta variant (B.1.351). Ferrets were exposed to droplets containing infectious SARS-CoV-2, ranging between 4 and 106 µm in diameter, simulating larger droplets produced by a cough from an infected person. Following exposure, viral RNA was detected on the fur of ferrets, and was deposited onto environmental surfaces, as well as the fur of ferrets placed in direct contact; SARS-CoV-2 remained infectious on the fur for at least 48 h. Low levels of viral RNA were detected in the nasal washes early post-exposure, yet none of the directly exposed, or direct-contact ferrets, became robustly infected or seroconverted to SARS-CoV-2. In comparison, ferrets intranasally inoculated with the SARS-CoV-2 beta variant became robustly infected, shedding viral RNA and infectious virus from the nasal cavity, with transmission to 75 % of naive ferrets placed in direct contact. These data suggest that larger infectious droplet nuclei and contaminated fur play minor roles in SARS-CoV-2 transmission among mustelids and potentially other companion animals.

Effect of limestone dose on the incidence of Rhizoctonia root rot and soybean yield in four soil types

AbstractRhizoctonia root rot (RRR) is a disease caused by the soilborne pathogen Rhizoctonia solani, which kills soybean plants in patches over croplands. Control is difficult because the pathogen survives in the soil and has a wide host range; therefore, cultural control methods might help to diminish disease pressure. The objective of this study was to evaluate liming as a strategy to reduce the incidence of RRR in soybean grown in four soil types: acrisol (Ac), cambisol Ta (CaTa), cambisol Tb (CaTb) and nitisol (Nit). The treatments were arranged in a factorial design of four doses of limestone (0, 3, 6 and 9 t/ha) at two sites within each field (inside and outside a Rhizoctonia patch). The incidence of RRR inside the patch was on average 40.3% whereas outside it was 1.2%. The reduction in disease incidence was 3.9%, 3.7%, 3.6% and 1.8% per 1000 kg of limestone applied for the Ac, CaTa, CaTb and Nit soils, respectively. The highest soybean yield was of 4713 and 3349 kg/ha for the Ac and Nit soils with application of 6.6 and 6.2 t/ha of limestone, respectively. Soybean yield in the cambisols increased linearly with liming, with an increase in yield of 121 and 172 kg/ha in CaTa and CaTb, respectively, for each 1000 kg of limestone applied. Thus, liming reduced the incidence of RRR in all soils and promoted better soybean nutrition, increasing soybean yield. Therefore, liming is a practice that contributes to both soil fertility and disease control.

Climate Change Promotes the Large-Scale Population Growth of Grapholita molesta (Busck) (Lepidoptera: Tortricidae) within Peach Orchards in China

Cosmopolitan agricultural herbivorous pests are provided with a wide range of potential hosts. Therefore, they have high carrying capacity, and can cause extremely severe damage in agroecosystems. Understanding the ecological mechanisms of their population dynamics, especially as they relate to large-scale meteorological variations and geographical landscape influences, can help us to reveal how they became such important pests. The oriental fruit moth, Grapholita molesta, is a typical example of a significant pest distributed on a large scale, which is capable of damaging fruit trees of economic value such as peach, apple, pear, etc. This pest not only occurs in China, but exists on all continents except Antarctica. In order to prevent major pests and diseases, a system of plant protection has been established gradually in peach orchards within the Modern Agro-industry Technology Research System in China (CARS) since 2009. In the system, we collected the monitoring data of G. molesta by using pheromone traps at 17 experimental stations, and then used the corresponding climate data (temperature and precipitation) to explore the link between climate factors using mixed models. The results show that both monthly mean temperature and precipitation had a significant positive correlation with the occurrence of G. molesta. Therefore, global warming with higher levels of precipitation may favor G. molesta, allowing it to outperform other potential pests at the population level in peach orchards, on a large scale.

Role of Goats in the Epidemiology of Coxiella burnetii.

Since its first description in the late 1930s, Q fever has raised many questions. Coxiella burnetii, the causative agent, is a zoonotic pathogen affecting a wide range of hosts. This airborne organism leads to an obligate, intracellular lifecycle, during which it multiplies in the mononuclear cells of the immune system and in the trophoblasts of the placenta in pregnant females. Although some issues about C. burnetii and its pathogenesis in animals remain unclear, over the years, some experimental studies on Q fever have been conducted in goats given their excretion pattern. Goats play an important role in the epidemiology and economics of C. burnetii infections, also being the focus of several epidemiological studies. Additionally, variants of the agent implicated in human long-term disease have been found circulating in goats. The purpose of this review is to summarize the latest research on C. burnetii infection and the role played by goats in the transmission of the infection to humans.

Facultative symbionts are potential agents of symbiont-mediated RNAi in aphids.

Aphids are major crop pests, and they can be controlled through the application of the promising RNA interference (RNAi) techniques. However, chemical synthesis yield of dsRNA for RNAi is low and costly. Another sustainable aphid pest control strategy takes advantage of symbiont-mediated RNAi (SMR), which can generate dsRNA by engineered microbes. Aphid host the obligate endosymbiont Buchnera aphidicola and various facultative symbionts that not only have a wide host range but are also vertically and horizontally transmitted. Thus, we described the potential of facultative symbionts in aphid pest control by SMR. We summarized the community and host range of these facultative symbionts, and then reviewed their probable horizontal transmitted routes and ecological functions. Moreover, recent advances in the cultivation and genetic engineering of aphid facultative symbionts were discussed. In addition, current legislation of dsRNA-based pest control strategies and their safety assessments were reviewed.

Symptomless turnip yellows virus infection causes grain yield loss in lentil and field pea: A three-year field study in south-eastern Australia.

Turnip yellows virus (TuYV) is a damaging virus that is persistently transmitted by aphids and infects a wide range of grain hosts including lentil (Lens culinaris Medik), field pea (Pisum sativum L.) and canola (Brassica napus L., oilseed rape). Although information is available about the effects of TuYV infection on grain yield in canola, data about its impact on yield in pulses is lacking. In this study, field experiments quantifying the effects of TuYV infection on the grain yield of lentil and field pea were conducted over three consecutive years (2018-2020) with varying weather conditions. Plants artificially inoculated with TuYV using viruliferous green peach aphid (Myzus persicae, Sulzer) were grown under typical field conditions in south-eastern Australia. At maturity, grain yield, along with associated grain and plant growth parameters, were measured. Compared to the non-inoculated control treatment, early TuYV infection reduced grain yield by up to 36% in lentil and 45% in field pea, while late TuYV infection had no significant impact on yield. Despite a high incidence of TuYV infection and significant yield losses recorded in inoculated plots, no obvious symptoms of virus infection were observed in the inoculated plots in any of the six experiments; this lack of visible symptoms in lentil and field pea has significant implications for crop health assessments, demonstrating the importance of testing for virus instead of relying solely on the presence of visual symptoms, and may also be leading to an underestimation of the importance of TuYV in pulses in Australia.