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At least 199 records · Page 11Linked to original sources

Susceptibility of ocean- and stream-type Chinook salmon to isolates of the L, U, and M genogroups of infectious hematopoietic necrosis virus (IHNV)

This study examined the susceptibility of Chinook salmon Oncorhynchus tshawytscha to viral strains from the L, U, and M genogroups of infectious hematopoietic necrosis virus (IHNV) present in western North America. The goal of this investigation was to establish a baseline understanding of the susceptibility of ocean- and stream-type Chinook salmon to infection and mortality caused by exposure to commonly detected strains of L, U, and M IHNV. The L IHNV strain tested here was highly infectious and virulent in both Chinook salmon populations, following patterns previously reported for Chinook salmon. Furthermore, ocean- and stream-type Chinook salmon fry at 1 g can also become subclinically infected with U and M strains of IHNV without experiencing significant mortality. The stream-type life history phenotype was generally more susceptible to infection and suffered greater mortality than the ocean-type phenotype. Between the U and M genogroup strains tested, the U group strains were generally more infectious than the M group strains in both Chinook salmon types. Substantial viral clearance occurred by 30 d post exposure, but persistent viral infection was observed with L, U, and M strains in both host populations. While mortality decreased with increased host size in stream-type Chinook salmon, infection prevalence was not lower for all strains at a greater size. These results suggest that Chinook salmon may serve as reservoirs and/or vectors of U and M genogroup IHNV.

Diseases of Aquatic Organisms↗

Comparative susceptibilities of selected California Chinook salmon and steelhead populations to isolates of L Genogroup Infectious Hematopoietic Necrosis Virus (IHNV)

Salmonid species demonstrate varied susceptibility to the viral pathogen infectious hematopoietic necrosis virus (IHNV). In California conservation hatcheries, juvenile Chinook salmon ( Oncorhynchus tshawytscha) have experienced disease outbreaks due to L genogroup IHNV since the 1940s, while indigenous steelhead (anadromous O. mykiss) appear relatively resistant. To characterize factors contributing to the losses of California salmonid fish due to IHNV, three populations of Chinook salmon and two populations of steelhead native to California watersheds were compared in controlled waterborne challenges with California L genogroup IHNV isolates at viral doses of 10 4 –10 6 pfu mL −1 . Chinook salmon fry were moderately to highly susceptible (CPM = 47–87%) when exposed to subgroup LI and LII IHNV. Susceptibility to mortality decreased with increasing age and also with a higher temperature. Mortality for steelhead fry exposed to two IHNV isolates was low (CPM = 1.3–33%). There was little intraspecies variation in susceptibility among populations of Chinook salmon and no differences in virulence between viruses strains. Viral persistence was demonstrated by the isolation of low levels of infectious IHNV from the skin of two juvenile Chinook salmon at 215 d post exposure. The persistence of the virus among Chinook salmon used for stocking into Lake Oroville may be an explanation for the severe epidemics of IHN at the Feather River hatchery in 1998–2002.

California↗

Temperature variation and host immunity regulate viral persistence in a salmonid host

Environmental variation has important effects on host–pathogen interactions, affecting large-scale ecological processes such as the severity and frequency of epidemics. However, less is known about how the environment interacts with host immunity to modulate virus fitness within hosts. Here, we studied the interaction between host immune responses and water temperature on the long-term persistence of a model vertebrate virus, infectious hematopoietic necrosis virus (IHNV) in steelhead trout ( Oncorhynchus mykiss ). We first used cell culture methods to factor out strong host immune responses, allowing us to test the effect of temperature on viral replication. We found that 15 ∘ "> ∘ C water temperature accelerated IHNV replication compared to the colder 10 and 8 ∘ "> ∘ C temperatures. We then conducted in vivo experiments to quantify the effect of 6, 10, and 15 ∘ "> ∘ C water temperatures on IHNV persistence over 8 months. Fish held at 15 and 10 ∘ "> ∘ C were found to have higher prevalence of neutralizing antibodies compared to fish held at 6 ∘ "> ∘ C. We found that IHNV persisted for a shorter time at warmer temperatures and resulted in an overall lower fish mortality compared to colder temperatures. These results support the hypothesis that temperature and host immune responses interact to modulate virus persistence within hosts. When immune responses were minimized (i.e., in vitro) virus replication was higher at warmer temperatures. However, with a full potential for host immune responses (i.e., in vivo experiments) longer virus persistence and higher long-term virulence was favored in colder temperatures. We also found that the viral RNA that persisted at later time points (179 and 270 days post-exposure) was mostly localized in the kidney and spleen tissues. These tissues are composed of hematopoietic cells that are favored targets of the virus. By partitioning the effect of temperature on host and pathogen responses, our results help to better understand environmental drivers of host–pathogen interactions within hosts, providing insights into potential host–pathogen responses to climate change.

Pathogens↗

Detection and genetic characterization of red-spotted grouper nervous necrosis virus and a novel genotype of nervous necrosis virus in black sea bass from the U.S. Atlantic coast

Nervous necrosis virus (NNV) causes a neurologic disease in a wide range of marine fish and poses serious disease risks to marine aquaculture worldwide. Little is known about the presence of NNV along the Atlantic coast of the United States, aside from the presence of barfin flounder nervous necrosis virus (BFNNV) in coldwater species in the northern part of this range. Herein we conducted surveillance for NNV from 2020 to 2022 in the mid-Atlantic region of the United States in black sea bass Centropristis striata , a serranid fish that is found throughout the eastern U.S. coast. Molecular detection methods have identified and characterized red-spotted grouper nervous necrosis virus (RGNNV) sequences at low prevalence throughout the years. Further, in 2022, a higher prevalence of a novel NNV genotype, tentatively named black sea bass nervous necrosis virus (BSBNNV), was characterized for the first time. Though virus isolation was unsuccessful, this study was the first to genetically identify NNV in this region and in this species. These findings highlight the need for further research on NNV to understand epidemiology and virulence in the context of marine fisheries and an emerging marine aquaculture industry in the United States.

New Jersey↗

Immunity to fish rhabdoviruses

Members of the family Rhabdoviridae are single-stranded RNA viruses and globally important pathogens of wild and cultured fish and thus relatively well studied in their respective hosts or other model systems. Here, we review the protective immune mechanisms that fish mount in response to rhabdovirus infections. Teleost fish possess the principal components of innate and adaptive immunity found in other vertebrates. Neutralizing antibodies are critical for long-term protection from fish rhabdoviruses, but several studies also indicate a role for cell-mediated immunity. Survival of acute rhabdoviral infection is also dependent on innate immunity, particularly the interferon (IFN) system that is rapidly induced in response to infection. Paradoxically, rhabdoviruses are sensitive to the effects of IFN but virulent rhabdoviruses can continue to replicate owing to the abilities of the matrix (M) protein to mediate host-cell shutoff and the non-virion (NV) protein to subvert programmed cell death and suppress functional IFN. While many basic features of the fish immune response to rhabdovirus infections are becoming better understood, much less is known about how factors in the environment affect the ecology of rhabdovirus infections in natural populations of aquatic animals.

Viruses↗

Procedures and rationale for marking sea otters captured and treated during the T/V Exxon Valdez oil spill

Four methods were used for marking sea otters ( Enhydra lutris ) captured and treated during the response to the T/V Exxon Valdez oil spill. Colored and numbered flipper tags were placed on each sea otter that was brought to the otter treatment centers. These tags allowed individual recognition and permitted the tracking of individuals through the treatment and holding processes. Recovery of tagged carcasses may provide a crude measure of the fates of rehabilitated otters. Seven sea otters were instrumented with radio transmitters attached to flipper tags as part of a pilot release program. The results of that study were inconclusive. Forty-five sea otters were implanted with radio transmitters as part of a study to assess the fate of rehabilitated sea otters. Specific objectives of the study include estimating survived rates and monitoring the reproductive success of the sample of rehabilitated sea otters and comparing the results with similar variables in a control population. Transponder chips were injected in the perianal region of all but seven of the instrumented sea otters, and all were tagged with red flipper tags. The advantages and disadvantages of each type of tag are discussed.

Alaska↗

Characterization of poxviruses from forest birds in Hawaii

Two strains of avian pox viruses were isolated from cutaneous lesions in Hawaiian crows ( Corvus hawaiiensis ) examined in 1994 and a third from a biopsy obtained in 1992 from an infected bird of the Apapane species ( Himatione sanguinea ) by inoculation of the chorioallantoic membranes (CAM) of developing chicken embryos. The resulting proliferative CAM lesions contained eosinophilic cytoplasmic inclusion bodies characteristic of pox virus infection. The pathogenicity of these three viruses in domestic chickens was mild as evidenced by the development of relatively minor lesions of short duration at the sites of inoculation. Their virulence in this host was similar to that of a fowlpox virus (FPV) vaccine strain and contrasted greatly with the ability of two field strains of FPV to produce extensive proliferative lesions. One of the Hawaiian crow pox virus isolates as well as the one originating from the Apapane species could be propagated in two secondary avian cell lines, QT-35 and LMH. A comparison of the restriction fragment length polymorphisms (RFLP) of the genomes of the two cell line-adapted viruses, generated by Eco RI digestion, revealed a limited degree of similarity. Moreover, neither profile was comparable to those of the two field isolates of FPV, which were almost indistinguishable from each other. Thus, based on the genetic distinctness of the two Hawaiian bird viruses, they appear to represent different strains of avipoxvirus.

Hawaii↗

Persistence of Pasteurella multocida in wetlands following avian cholera outbreaks

Avian cholera, caused by Pasteurella multocida, affects waterbirds across North America and occurs worldwide among various avian species. Once an epizootic begins, contamination of the wetland environment likely facilitates the transmission of P. multocida to susceptible birds. To evaluate the ability of P. multocida serotype-1, the most common serotype associated with avian cholera in waterfowl in western and central North America, to persist in wetlands and to identify environmental factors associated with its persistence, we collected water and sediment samples from 23 wetlands during winters and springs of 1996a??99. These samples were collected during avian cholera outbreaks and for up to 13 wk following initial sampling. We recovered P. multocida from six wetlands that were sampled following the initial outbreaks, but no P. multocida was isolated later than 7 wk after the initial outbreak sampling. We found no significant relationship between the probability of recovery of P. multocida during resampling and the abundance of the bacterium recovered during initial sampling, the substrate from which isolates were collected, isolate virulence, or water quality conditions previously suggested to be related to the abundance or survival of P. multocida. Our results indicate that wetlands are unlikely to serve as a long-term reservoir for P. multocida because the bacterium does not persist in wetlands for long time periods following avian cholera outbreaks.

Journal of Wildlife Diseases↗

Using amplified fragment length polymorphism analysis to differentiate isolates of Pasteurella multocida serotype 1

Avian cholera, an infectious disease caused by the bacterium Pasteurella multocida , kills thousands of North American wild waterfowl annually. Pasteurella multocida serotype 1 isolates cultured during a laboratory challenge study of Mallards ( Anas platyrhynchos ) and collected from wild birds and environmental samples during avian cholera outbreaks were characterized using amplified fragment length polymorphism (AFLP) analysis, a whole-genome DNA fingerprinting technique. Comparison of the AFLP profiles of 53 isolates from the laboratory challenge demonstrated that P. multocida underwent genetic changes during a 3-mo period. Analysis of 120 P. multocida serotype 1 isolates collected from wild birds and environmental samples revealed that isolates were distinguishable from one another based on regional and temporal genetic characteristics. Thus, AFLP analysis had the ability to distinguish P. multocida isolates of the same serotype by detecting spatiotemporal genetic changes and provides a tool to advance the study of avian cholera epidemiology. Further application of AFLP technology to the examination of wild bird avian cholera outbreaks may facilitate more effective management of this disease by providing the potential to investigate correlations between virulence and P. multocida genotypes, to identify affiliations between bird species and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission.

California, Iowa, Missouri, Nebraska, Utah, Wiscon↗

Immunization of black-tailed prairie dog against plague through consumption of vaccine-laden baits

Prairie dogs ( Cynomys spp.) are highly susceptible to Yersinia pestis and, along with other wild rodents, are significant reservoirs of plague for other wildlife and humans in the western United States. A recombinant raccoon poxvirus, expressing the F1 antigen of Y. pestis , was incorporated into a palatable bait and offered to three groups ( n =18, 19, and 20) of black-tailed prairie dogs ( Cynomys ludovicianus ) for voluntary consumption, either one, two, or three times, at roughly 3-wk intervals. A control group ( n =19) received baits containing raccoon poxvirus without the inserted antigen. Mean antibody titers to Y. pestis F1 antigen increased significantly in all groups ingesting the vaccine-laden baits, whereas the control group remained negative. Upon challenge with virulent Y. pestis , immunized groups had higher survival rates (38%) than the unimmunized control group (11%). The mean survival time of groups ingesting vaccine-laden baits either two or three times was significantly higher than that of animals ingesting vaccine-laden baits just one time and of animals in the control group. These results show that oral immunization of prairie dogs against plague provides some protection against challenge at dosages that simulate simultaneous delivery of the plague bacterium by numerous (3–10) flea bites.

Journal of Wildlife Diseases↗

The innate immune response may be important for surviving plague in wild Gunnison's prairie dogs

Prairie dogs (Cynomys spp.) are highly susceptible to Yersinia pestis, with ≥99% mortality reported from multiple studies of plague epizootics. A colony of Gunnison's prairie dogs (Cynomys gunnisoni) in the Aubrey Valley (AV) of northern Arizona appears to have survived several regional epizootics of plague, whereas nearby colonies have been severely affected by Y. pestis. To examine potential mechanisms accounting for survival in the AV colony, we conducted a laboratory Y. pestis challenge experiment on 60 wild-caught prairie dogs from AV and from a nearby, large colony with frequent past outbreaks of plague, Espee (n = 30 per colony). Test animals were challenged subcutaneously with the fully virulent Y. pestis strain CO92 at three doses: 50, 5,000, and 50,000 colony-forming units (cfu); this range is lethal in black-tailed prairie dogs (Cynomys ludovicianus). Contrary to our expectations, only 40% of the animals died. Although mortality trended higher in the Espee colony (50%) compared with AV (30%), the differences among infectious doses were not statistically significant. Only 39% of the survivors developed moderate to high antibody levels to Y. pestis, indicating that mechanisms other than humoral immunity are important in resistance to plague. The ratio of neutrophils to lymphocytes was not correlated with plague survival in this study. However, several immune proteins with roles in innate immunity (VCAM-1, CXCL-1, and vWF) were upregulated during plague infection and warrant further inquiry into their role for protection against this disease. These results suggest plague resistance exists in wild populations of the Gunnison's prairie dog and provide important directions for future studies.

Arizona↗

Retrospective analysis of the epidemiologic literature, 1990–2015, on wildlife-associated diseases from the Republic of Korea

To assess the status of research on wildlife diseases in the Republic of Korea (ROK) and to identify trends, knowledge gaps, and directions for future research, we reviewed epidemiologic publications on wildlife-associated diseases in the ROK. We identified a relatively small but rapidly increasing body of literature. The majority of publications were focused on public or livestock health and relatively few addressed wildlife health. Most studies that focused on human and livestock health were cross-sectional whereas wildlife health studies were mostly case reports. Fifteen diseases notifiable to the World Organisation for Animal Health were identified and 21 diseases were identified as notifiable to either the Korean Ministry of Health, Welfare, and Family Affairs or the Korean Ministry of Agriculture. Two diseases were reported as occurring as epidemics; highly pathogenic avian influenza (HPAI) and virulent Newcastle disease. Six diseases or disease agents were described in the literature as emerging including HPAI, rabies, Babesia microti , avian coronaviruses, scrub typhus, and severe fever thrombocytopenia syndrome virus. The diseases for which there were the largest number of publications were HPAI and rabies. The majority of wildlife-associated zoonotic disease publications focused on food-borne parasitic infections or rodent-associated diseases. Several publications focused on the potential of wildlife as reservoirs of livestock diseases; in particular, water deer ( Hydropotes inermis ) and wild boar ( Sus scrofa ). In contrast, there were few publications on diseases of concern for wildlife populations or research to understand the impacts of these diseases for wildlife management. Increased focus on prospective studies would enhance understanding of disease dynamics in wildlife populations. For the high-consequence diseases that impact multiple sectors, a One Health approach, with coordination among the public health, agricultural, and environmental sectors, would be important. This type of review can provide useful information for countries or regions planning or implementing national wildlife health programs.

Journal of Wildlife Diseases↗

Application of diagnostic tests for mycoplasmal infections of desert and gopher tortoises with management recommendations

Mycoplasmosis is a transmissible upper respiratory tract disease that has affected plans for management and conservation of wild desert and gopher tortoises in the United States. Although impact of mycoplasmosis on populations of desert and gopher tortoises is unknown, increased prevalence of seropositive animals as well as field observations of clinically ill tortoises have occurred in association with declining populations. In order to help in the identification of potentially infected animals, three tests have been developed to diagnose mycoplasmal infections of tortoises: 1) direct mycoplasmal culture; 2) detection of mycolplasmal chromosomal DNA by polymerase chain reaction (PCR); and 3) detection of anti-Mycoplasma antibodies in tortoise plasma by enzyme-linked immunosorbent assay (ELISA). Each test provides different, complementary information that collectively can be used to define tortoise mycoplasmal infection status. The types of samples required, the predictive value, interpretation, and cost vary among tests. These assays have been used for epidemiological surveys and in decision making for relocation, repatriation, or captive management of tortoises to minimize the risk of outbreaks of mycoplasmal respiratory disease and spread of the causative agent of this disease. Certain features of mycoplasmal infections of tortoises and other animals create a diagnostic dilemma. Multiple Mycoplasma species can cause respiratory disease with identical clinical presentations. Further, individual strains of a given species may vary with respect to their virulence potential, and some species may be commensals rather than pathogens. Current diagnostic tests may not differentiate among mycoplasmal species or strains or permit determination of pathogenicity of individual isolates. Thus, the information provided by testing is not a simple 'positive' vs. 'negative' issue. While these tests provide much needed information on the exposure of tortoise populations to Mycoplasma species, they do not provide a complete picture of the overall health status of individual tortoises or populations. Unfortunately, test results are often used to make life and death decisions concerning disposition of tortoises being displaced by land development without a complete understanding of the limitations of the diagnostic tests or any consideration of other infectious agents that might be present.

AZ, CA, FL, NV, UT↗

Multi-species patterns of avian cholera mortality in Nebraska's rainwater basin

Nebraska's Rainwater Basin (RWB) is a key spring migration area for millions of waterfowl and other avian species. Avian cholera has been endemic in the RWB since the 1970s and in some years tens of thousands of waterfowl have died from the disease. We evaluated patterns of avian cholera mortality in waterfowl species using the RWB during the last quarter of the 20th century. Mortality patterns changed between the years before (1976 - 1988) and coincident with (1989 - 1999) the dramatic increases in lesser snow goose abundance and mortality. Lesser snow geese (Chen caerulescens caerulescens) have commonly been associated with mortality events in the RWB and are known to carry virulent strains of Pasteurella multocida, the agent causing avian cholera. Lesser snow geese appeared to be the species most affected by avian cholera during 1989 - 1999; however, mortality in several other waterfowl species was positively correlated with lesser snow goose mortality. Coincident with increased lesser snow goose mortality, spring avian cholera outbreaks were detected earlier and ended earlier compared to 1976 - 1988. Dense concentrations of lesser snow geese may facilitate intraspecific disease transmission through bird-to-bird contact and wetland contamination. Rates of interspecific avian cholera transmission within the waterfowl community, however, are difficult to determine.

Nebraska↗

Spatial dynamics of bar-headed geese migration in the context of H5N1

Virulent outbreaks of highly pathogenic avian influenza (HPAI) since 2005 have raised the question about the roles of migratory and wild birds in the transmission of HPAI. Despite increased monitoring, the role of wild waterfowl as the primary source of the highly pathogenic H5N1 has not been clearly established. The impact of outbreaks of HPAI among species of wild birds which are already endangered can nevertheless have devastating consequences for the local and non-local ecology where migratory species are established. Understanding the entangled dynamics of migration and the disease dynamics will be key to prevention and control measures for humans, migratory birds and poultry. Here, we present a spatial dynamic model of seasonal migration derived from first principles and linking the local dynamics during migratory stopovers to the larger scale migratory routes. We discuss the effect of repeated epizootic at specific migratory stopovers for bar-headed geese ( Anser indicus ). We find that repeated deadly outbreaks of H5N1 on stopovers during the autumn migration of bar-headed geese could lead to a larger reduction in the size of the equilibrium bird population compared with that obtained after repeated outbreaks during the spring migration. However, the opposite is true during the first few years of transition to such an equilibrium. The age-maturation process of juvenile birds which are more susceptible to H5N1 reinforces this result.

Journal of the Royal Society Interface↗

Multi-state analysis of the impacts of avian pox on a population of Serins (Serinus serinus): The importance of estimating recapture rates

Disease is one of the evolutionary forces shaping populations. Recent studies have shown that epidemics like avian pox, malaria, or mycoplasmosis have affected passerine population dynamics, being responsible for the decline of some populations or disproportionately killing males and larger individuals and thus selecting for specific morphotypes. However, few studies have estimated the effects of an epidemic by following individual birds using the capture-recapture approach. Because avian pox can be diagnosed by direct examination of the birds, we are here able to analyze, using multistate models, the development and consequences of an avian pox epidemic affecting in 1996, a population of Serins (Serinus serinus) in northeastern Spain. The epidemics lasted from June to the end of November of 1996, with a maximum apparent prevalence rate > 30% in October. However, recapture rate of sick birds was very high (0.81, range 0.37-0.93) compared to that of healthy birds (0.21, range 0.020-32), which highly inflated apparent prevalence rate. This was additionally supported by the low predicted transition from the state of being uninfected to the state of being infected (0.03, SE 0.03). Once infected, Serin avian pox was very virulent with (15-day) survival rate of infected birds being of only 0.46 (SE 0.17) compared to that of healthy ones (0.87, SE 0.03). Probability of recovery from disease, provided that the bird survived the first two weeks, however, was very high (0.65, SE 0.25). The use of these estimates together with a simple model, allowed us to predict an asymptotic increase to prevalence of about 4% by the end of the outbreak period, followed by a sharp decline, with the only remaining infestations being infected birds that had not yet recovered. This is in contrast to the apparent prevalence of pox and stresses the need to estimate recapture rates when estimating population dynamics parameters. ?? 2004 Museu de Cie??ncies Naturals.

Animal Biodiversity and Conservation↗

Economic analysis of the proposed rule to prevent arrival of new genetic strains of the rust fungus Puccinia psidii in Hawai'i.

Since its first documented introduction to Hawai‘i in 2005, the rust fungus P. psidii has already severely damaged Syzygium jambos (Indian rose apple) trees and the federally endangered Eugenia koolauensis (nioi). Fortunately, the particular strain has yet to cause serious damage to ‘ōhi‘a, which comprises roughly 80% of the state’s native forests and covers 400,000 ha. Although the rust has affected less than 5% of Hawaii’s ‘ōhi‘a trees thus far, the introduction of more virulent strains and the genetic evolution of the current strain are still possible. Since the primary pathway of introduction is Myrtaceae plant material imported from outside the state, potential damage to ‘ohi‘a can be minimized by regulating those high-risk imports. We discuss the economic impact on the state’s florist, nursery, landscaping, and forest plantation industries of a proposed rule that would ban the import of non-seed Myrtaceae plant material and require a one-year quarantine of seeds. Our analysis suggests that the benefits to the forest plantation industry of a complete ban on non-seed material would likely outweigh the costs to other affected sectors, even without considering the reduction in risk to ‘ōhi‘a. Incorporating the value of ‘ōhi‘a protection would further increase the benefit-cost ratio in favor of an import ban.

Hawaii↗

Field performance of cucurbit and tomato plants infected with a nonpathogenic mutant of Colletotrichum magna (teleomorph: Glomerella magna; Jekins and Winstead)

Path-1 is a UV-induced non-pathogenic mutant of a virulent Colletotrichum magna isolate that establishes mutualistic symbioses with cucurbit and tomato species. Under laboratory conditions, this mutualism results in plant growth enhancement, drought tolerance, and disease protection against fungal pathogens. This study focuses on the efficacy of this symbiosis and the symbiotic lifestyle expressed by path-1 under field conditions in the absence of disease stress. The effects of colonization by path-1 on fruit yields and growth was measured in field plots with four cucurbit species including four watermelon cultivars, and two tomato cultivars, over four growing seasons. The persistence of the symbiosis, extent of colonization, and path-1 transmission were also assessed. Yields from path-1 infected plants were equivalent to or greater than yields from non-inoculated control plants and path-1 systemically colonized plants throughout each growing season. Path-1 also increased the growth rates of tomato plants and was not transmitted to uncolonized plants. The results indicate that there are no metabolic costs of this symbiosis and the symbiosis is maintained under field conditions.

Symbiosis↗