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A summary of information on the rust Puccinia psidii Winter (guava rust) with emphasis on means to prevent introduction of additional strains to Hawaii

The neotropical rust fungus Puccinia psidii ( P. psidii ) was originally described from the host common guava in its native Brazil but has been found since on hosts throughout the myrtle family (Myrtaceae), including a dramatic host jump to nonnative Eucalyptus plantations. Most rust fungi are able to live only on a very narrow range of host species. P. psidii is unusual both for having a broad host range and for the intensity of its damage to susceptible young growth. This rust first got a foothold in the United States in Florida more than three decades ago. The U.S. Department of Agriculture (USDA) has since considered it a nonactionable, nonreportable pest. Hawaii and Florida are the only two states with native species in the myrtle family. Over a period of 30 years, this rust has done little damage to any of the scattered native Myrtaceae in Florida, although the host range of the rust has gradually grown to about 30 mostly nonnative species in the family, apparently because of increasing genetic variety of the rust by repeated introductions. However, Florida’s native Myrtaceae are among the roughly 1,100 neotropical species that are largely resistant to P. psidii . The 3,000 species of non-neotropical Myrtaceae of the Pacific, Australia, Asia, and Africa are expected to prove much more vulnerable to P. psidii . Little is known about the genetics or genetic strains of P. psidii , although existing literature shows that there are numerous strains that have differential ability to infect suites of host plants. The rust was first recorded in the state of Hawaii on Oahu in April 2005 and quickly spread throughout the Hawaiian Islands. The main concern in Hawaii became the potential threat to ohia, Metrosideros polymorpha (Myrtaceae), the endemic forest tree species overwhelmingly important in Hawaii’s nature and culture. The potential ecological consequences of a virulent strain of rust on ohia forests are immense, due to its role as a foundation tree species and the diversity of niches it fills in Hawaii. A single genetic strain of the rust is established in Hawaii, apparently composed of a single genotype lacking sexual reproduction. P. psidii has been found statewide in Hawaii attacking Myrtaceae from near sea level to about 1,200 m elevation in areas with rainfall ranging from 750–5,000 mm. Five of eight native Myrtaceae and at least 15 nonnative species have been observed as hosts of P. psidii in Hawaii. The federally endangered Eugenia koolauensis (nioi) and the nonendangered indigenous species Eugenia reinwardtiana are severely damaged. The introduced (an Asian species) and invasive rose apple, Syzygium jambos , is severely affected at a landscape scale, with widespread crown dieback and many instances of complete tree death. In spite of billions of wind-dispersed rust spores produced from rose apple infestations during 2006 to 2008, adjacent ohia have been little affected to date by the rust strain in Hawaii. Within the elevation range of the rust, P. psidii is found on less than 5 percent of the ohia trees in the wild; on those ohia trees on which the rust is found, it is normally found on less than 5 percent of the leaves. The strain in Hawaii has not attacked many of the species known to be infected by the rust elsewhere, including common guava. On the basis of the very substantial genetic diversity of the much-studied, crop-damaging species of the genus Puccinia , there is good reason to believe that there are at minimum dozens and likely hundreds or thousands of genotypes of P. psidii , likely concentrated in the core range in Brazil but with potential for dispersal by globalization. Multiple genotypes are believed already present in the United States and certain to spread freely in the absence of restrictions. The U.S. Forest Service has initiated a major collaborative project in Brazil to investigate the genetics of susceptibility of Hawaii’s ohia to P. psidii , but initial results will likely not be available for several years. If just one more strain reaches Hawaii, the consequences could be dire for ohia, with each new genotype arriving having an unknown likelihood of increasing damage to ohia; possibilities for mutation and (or) genetic mixing, even with asexual strains, are apparently substantial, based on what is known about other Puccinia species. Investigations are needed to clarify rust-nioi relationships. However, it is likely that keeping out new strains of P. psidii may be important for long-term survival of nioi as well as for the health of ohia forest. The source of Hawaii’s initial invasion by P. psidii is uncertain but is strongly suspected to have been decorative foliage of species in the myrtle family from the mainland United States, most likely California, where there had been outbreaks of this rust on cultivated myrtle in 2005. In 2006–7, Maui’s Hawaii Department of Agriculture (HDOA) inspectors intercepted several P. psidii infected shipments of foliage myrtle, shipped from several California counties. Recognizing the huge threat of the rust to Hawaii’s one million acres of ohia forests, and consequently to Hawaii’s watersheds and biodiversity, Hawaii’s Board of Agriculture unanimously approved an interim rule in August 2007 banning importation of plants in the myrtle family from “infested areas,” specified as South America, Florida, and California. However, the interim rule has not been made permanent by HDOA, and the department has stated that it needs further information to formulate a long-term rule that imposes appropriate measures. Rust spores can survive for 2 to 3 months, and the pathogen can be transported to Hawaii on Myrtaceae from anywhere in the world through the United States mainland. There is much geographic reshuffling of flowers and foliage among the far-flung firms in the trade, especially for bouquet making. Because P. psidii is a nonactionable and nonreportable pest in the United States, foliage and flowers of the myrtle family can move freely into the country (usually but not necessarily always through the ports of Miami or Los Angeles), and from state to state. Currently, the State of Hawaii regulates incoming plant material in the family Myrtaceae by visual inspection. Inspection capacity and latent (asymptomatic) infections limit the ability to detect the rust. New molecular tests could improve detection efficiency, but the cost and the time required to process samples currently precludes their routine use in ports of entry. Interdiction, which has effectively kept coffee rust ( Hemileia vastatrix ) out of Hawaii for 120 years, offers the strongest protection for Hawaii’s native ecosystems from P. psidii . Interdiction of Myrtaceae from the continental United States could have the important supplementary benefit of preventing establishment in Hawaii of other very significant pests of multiple species of Myrtaceae that are already in the country, including: the Eugenia psyllid Trioza eugeniae (Hemiptera: Psyllidae); Chrysophtharta m-fuscum , the Eucalyptus tortoise beetle (Coleoptera: Chrysomelidae); Leptocybe invasa , the blue gum chalcid wasp (Hymenoptera: Chalcidae); and the fungal pathogens Mycosphaerella molleriana (Ascomycota: Mycosphaerelliaceae, crinkle leaf disease of Eucalyptus spp.) and Neofusicoccum parvum (Ascomycota: Botryosphaeriaceae), currently causing serious damage to Syzygium paniculatum in south Florida nurseries. Each of these pests would be likely to cause very significant damage to native and (or) cultivated Myrtaceae in Hawaii. Each of these pests is a prime candidate for transport by the foliage and (or) nursery stock pathways from Florida and California into Hawaii. Hawaii Department of Agriculture has a clear mandate to protect Hawaii’s natural environment, forestry and cultivated Myrtaceae. Principles of the World Trade Organization’s Treaty on Sanitary and Phytosanitary Measures and the International Plant Protection Convention are consistent with the right of Hawaii to take action. The current threat of P. psidii and the other five serious threats to Myrtaceae are primarily posed by the importation of infected plants from the continental United States; however, that may change in the future. If Hawaii were to decide to take a stand (through State regulation) to protect its native and introduced Myrtaceae, there is a possibility that USDA would consider Federal regulation of Myrtaceae from foreign countries.

Hawaii

Black abalone surveys at Naval Base Ventura County, San Nicolas Island, California: 2019, annual report

The U.S. Geological Survey Western Ecological Research Center’s Santa Cruz Field Station, Santa Cruz, California, has been funded by the U.S. Navy to continue monitoring a suite of intertidal black abalone sites at San Nicolas Island, California. The nine rocky intertidal sites were established in 1980 by Glenn VanBlaricom (then of the U.S. Fish and Wildlife Service) to study the potential impact of translocated sea otters on the intertidal black abalone population at the island. The sites were monitored from 1981 to 1997, usually annually or semi-annually. Monitoring resumed in 2001, and regular annual monitoring cycles have been conducted at the sites since then. The study sites became particularly important, from a management perspective, after a virulent disease decimated black abalone populations throughout southern California beginning in the mid-1980s. The disease, withering syndrome, was first observed on San Nicolas Island in 1992 and during the next few years reduced the population there by approximately 99 percent. The species was subsequently listed as endangered under the Endangered Species Act in 2009. The subject of this report is the 2019 monitoring cycle of the sites and how the current status fits into the long-term data at San Nicolas Island. Since 2001, the monitored population has increased nearly tenfold to approximately 8.7 percent of the pre-disease level. This increase has resulted from generally higher levels of recruitment than seen in the first two decades of monitoring, punctuated by a few high recruitment events. Most of the population growth has been at two of the nine sites (sites 7 and 8). This pattern continued in 2019 with increasing numbers at sites 7 and 8 and the highest number of abalone counted and measured island-wide since 1996. However, counts declined at six of the sites during the last year and the increases in counts at sites 7 and 8 barely offset these losses. Recruitment rates have fallen since a peak in 2017 but 2019 continued to show some additional recruitment. The distance between adjacent black abalone, a metric relevant to potential reproduction, has decreased substantially since it was first consistently measured in 2005. Although sand burial can have devastating localized consequences to black abalone, the sand cover data we collected was not sufficient to suggest an obvious temporal or site-based pattern to sedimentation, and there is no indication that this was a factor in any of the declines recorded in 2019. Continued monitoring of these sites can provide island biologists with species trends to aid in adaptive management of the resource.

California

Black abalone surveys at Naval Base Ventura County, San Nicolas Island, California—2020, annual report

The U.S. Geological Survey monitors a suite of intertidal black abalone sites at San Nicolas Island, California, in cooperation with the U.S. Navy, which owns the island. The nine rocky intertidal sites were established in 1980 to study the potential impact of translocated sea otters on the intertidal black abalone population at the island. The sites were monitored from 1981 to 1997, usually annually or biennially. Monitoring resumed in 2001 and has been completed annually since then. At the time of this report, the work is conducted by the Western Ecological Research Center’s Santa Cruz Field Station, Santa Cruz, California. The study sites became particularly important, from a management perspective, after a virulent disease decimated black abalone populations throughout southern California beginning in the mid-1980s. The disease, withering syndrome, was first observed on San Nicolas Island in 1992 and during the next few years, it reduced the population there by more than 99 percent. The species was subsequently listed as endangered under the Endangered Species Act in 2009. The subject of this report is the 2020 monitoring cycle of the sites and how the current status fits into the long-term data at San Nicolas Island. Since 2001, the monitored population has increased twelvefold to approximately 9.6 percent of the pre-disease level. This increase has resulted from generally higher levels of recruitment than seen in the first two decades of monitoring, punctuated by a few unexplained high recruitment events. Most of the population growth has been at two of the nine sites (sites 7 and 8). This pattern continued in 2020, but with increasing numbers at all sites and the highest number of abalone counted and measured island-wide since 1993. Recruitment rates have fallen since a peak in 2017, but 2020 continued to show moderate levels of additional recruitment. The distance between adjacent black abalone has decreased substantially since it was first consistently measured in 2005, potentially indicating that the abalone are close enough to one another to reproduce successfully. Sand burial can have devastating localized consequences to black abalone, but there is evidence suggesting that they may be able to escape periodic sand inundation if suitable refugia exist. These data suggest that monitoring can inform adaptive management of the resource by base resource managers.

California

Black Abalone surveys at Naval Base Ventura County, San Nicolas Island, California—2021, annual report

The U.S. Geological Survey monitors a suite of intertidal black abalone sites at San Nicolas Island, California, in cooperation with the U.S. Navy, which owns the island. The nine rocky intertidal sites were established in 1980 to study the potential effect of translocated sea otters on the intertidal black abalone population at the island. The sites were monitored from 1981 to 1997, usually annually or biennially. Monitoring resumed in 2001 and has been completed annually since then. Since 2018, the work has been carried out by the U.S. Geological Survey Western Ecological Research Center. The study sites became particularly important, from a management perspective, after a virulent disease decimated black abalone populations throughout southern California beginning in the mid-1980s. The disease, withering syndrome, was first observed on San Nicolas Island in 1992 and during the next few years, it reduced the population there by more than 99 percent. The species was subsequently listed as endangered under the Endangered Species Act in 2009. The subject of this report is the 2021 survey of the sites and how the measured population status compares to the long-term data (collected over several decades) at San Nicolas Island. During the last two decades, the total monitored black abalone population at the island has grown approximately ten-fold after the disease related decline, from about 200 to more than 2,000 abalone. Since it was first consistently measured in 2005, the mean distance between adjacent black abalone has decreased substantially from approximately 50 centimeters to less than 15 centimeters, indicating that abalone are close enough together at several of the sites to reproduce successfully. The 2021 counts were the first since 2016 to indicate a possible decline in the monitored population at San Nicolas Island. Although still more than ten times the population counted in 2001, counts on the transects dropped by 13.6 percent from the survey count in 2020. The most significant decline was the loss of 341 abalone, from the previous count of 547, on a transect that since 2002 had the highest count of all 44 transects. Between 2020 and 2021, there were increases and decreases at the sites and the transects at each site. Although the 2021 count was lower than the 2020 count, it was the second highest since 1997. Based on the number of small abalone counted, recruitment rates were similar to most years since 2008 and higher than the rates observed before the population declines resulting from withering syndrome.

California

Black abalone surveys at Naval Base Ventura County, San Nicolas Island, California—2022 annual report

The U.S. Geological Survey monitors a suite of intertidal black abalone ( Haliotis cracherodii ) sites at San Nicolas Island, California, in cooperation with the U.S. Navy, which owns the island. The nine rocky intertidal sites were established in 1980 to study the potential effect of translocated southern sea otters ( Enhydra lutris nereis ) on the intertidal black abalone population at San Nicolas Island. The sites were monitored, typically annually or biennially, from 1981 to 1997. Monitoring resumed in 2001 and has been completed annually thereafter. Since 2018, the monitoring has been carried out by the U.S. Geological Survey Western Ecological Research Center. The study sites became particularly important from a management perspective after a virulent disease decimated black abalone populations throughout southern California beginning in the mid-1980s. The disease, withering syndrome, was first observed on San Nicolas Island in 1992, and during the next few years, withering syndrome reduced the black abalone population on San Nicolas Island by more than 99 percent. The black abalone was subsequently listed as endangered under the Endangered Species Act in 2009. The subject of this report is the 2022 survey of the sites and the status of the measured population of black abalone in comparison to long-term patterns (based on data collected since 1981) at San Nicolas Island. Between the years 2000 and 2022, the total monitored black abalone population on the island has grown from roughly 200 to more than 2,000, approximately a ten-fold increase following the disease-related decline. Since it was first consistently measured in 2005, the distance between adjacent black abalone has decreased substantially from approximately 50 centimeters to less than 15 centimeters, indicating that black abalone are sufficiently close together at several of the sites to reproduce successfully. The total black abalone count in 2022 was 2,156, which was 7.9 percent lower than the total count in 2020 but 6.6 percent higher than in 2021. There were increases and decreases among the sites and transects within each site in 2022, but six of the nine sites had higher counts than in the previous year. The 2022 count is one of the highest since 1993, second only to the 2020 count. In 2022, the annual recruitment rate, defined as the percentage of measured black abalone with a shell length of 3 centimeters or less, was the second highest recorded, only slightly less than in 2017.

California

Black abalone surveys at Naval Base Ventura County, San Nicolas Island, California—2023 annual report

The U.S. Geological Survey monitors a suite of intertidal black abalone ( Haliotis cracherodii ) sites at San Nicolas Island, California, in cooperation with the U.S. Navy, which owns the island. The nine rocky intertidal sites were established in 1980 to study the potential effect of translocated sea otters on the intertidal black abalone population at the island. The sites were monitored from 1981 to 1997, typically annually or biennially. Monitoring resumed in 2001 and has been completed annually thereafter. Since 2018, the work has been carried out by the U.S. Geological Survey Western Ecological Research Center. The study sites became particularly important, from a management perspective, after a virulent disease decimated black abalone populations throughout southern California beginning in the mid-1980s. The disease, withering syndrome ( Candidatus Xenohaliotis californiensis ), was first observed on San Nicolas Island in 1992 and over the next few years, withering syndrome reduced the black abalone population on San Nicolas Island by more than 99 percent. In 2009, the black abalone subsequently was listed as endangered under the Endangered Species Act. The subject of this report is the 2023 survey of the sites and the status of the measured population in comparison to long-term patterns (based on data collected since 1981) at San Nicolas Island. Between the years 2000 and 2023, the total monitored black abalone population at the island has grown from roughly 200 to more than 2,500 abalone following disease-related decline. Since it was first consistently measured in 2005, the average distance between adjacent black abalone has decreased substantially from approximately 50 centimeters to less than 15 centimeters, indicating that abalone are sufficiently close together at several of the sites to reproduce successfully. The total abalone count in 2023 was 2,570, which was 19.2 percent higher than in 2022 and the highest count since 1993. All nine sites had higher counts in 2023 than in the previous year. Over 25 percent of the black abalone counted in 2023 were classified as recruits, defined as having a shell length of 3 centimeters or less.

California

Pathogenic bacteria and microbial-source tracking markers in Brandywine Creek Basin, Pennsylvania and Delaware, 2009-10

The City of Wilmington, Delaware, is in the downstream part of the Brandywine Creek Basin, on the main stem of Brandywine Creek. Wilmington uses this stream, which drains a mixed-land-use area upstream, for its main drinking-water supply. Because the stream is used for drinking water, Wilmington is in need of information about the occurrence and distribution of specific fecally derived pathogenic bacteria (disease-causing bacteria) and their relations to commonly measured fecal-indicator bacteria (FIB), as well as information regarding the potential sources of the fecal pollution and pathogens in the basin. This study focused on five routinely sampled sites within the basin, one each on the West Branch and the East Branch of Brandywine Creek and at three on the main stem below the confluence of the West and East Branches. These sites were sampled monthly for 1 year. Targeted event samples were collected on two occasions during high flow and two occasions during normal flow. On the basis of this study, high flows in the Brandywine Creek Basin were related to increases in FIB densities, and in the frequency of selected pathogen and source markers, in the West Branch and main stem of Brandywine Creek, but not in the East Branch. Water exceeding the moderate fullbody-contact single-sample recreational water-quality criteria (RWQC) for Escherichia coli (E. coli) was more likely to contain selected markers for pathogenic E. coli ( eaeA , stx 1 , and rfb O157 gene markers) and bovine fecal sources ( E. hirae and LTIIa gene markers), whereas samples exceeding the enterococci RWQC were more likely to contain the same pathogenic markers but also were more likely to carry a marker indicative of human source ( esp gene marker). On four sample dates, during high flow between October and March, the West Branch was the only observed potential contributor of selected pathogen and bovine source markers to the main stem of Brandywine Creek. Indeed, the stx 2 marker, which indicates a highly virulent type of pathogenic E. coli , was found only in the West Branch and main stem at high flow but was not found in the East Branch under similar conditions. However, it must be noted that throughout the entire year of sampling there were occasions, during both high and normal flows, when both the East and West Branches were potential contributors of pathogen and microbial-source tracking markers to the main stem. Therefore, this study indicates that under selected conditions (high flow, October through March), West Branch Brandywine Creek Basin was the most likely source of elevated FIB densities in the main stem. These elevated densities are associated with more frequent detection of selected pathogenic E. coli markers ( rfb O157 stx 1 ) and are associated with MST markers of bovine source. However, during other times of the year, both the West Branch and East Branch Basins are acting as potential sources of FIB and fecally derived pathogens.

Pennsylvania, Delaware

Preliminary investigation of groundwater quality near a Michigan cemetery, 2016–17

The potential effect of cemetery leachate on groundwater quality in the United States has rarely been studied. Nutrients and other constituents associated with decomposition and burial processes (such as embalming) have the potential to reach shallow groundwater and could affect nearby drinking-water sources. The objective of this preliminary investigation was to evaluate the potential effect of cemetery leachate on shallow groundwater quality near Mt. Hope Cemetery in Ingham County, Lansing, Michigan, which is within the Well-head Protection Area for the City of Lansing. The constituents measured in this study include nutrients, trace metals, formaldehyde, fecal indicator bacteria, bacterial pathogen genes, contaminants of emerging concern (including pharmaceuticals, personal care products, and wastewater indicator compounds), and age-dating compounds. Three monitoring wells were installed 7 to 12 feet below land surface downgradient from the cemetery and sampled quarterly for 1 year. A fourth well (Fenner) was sampled to determine groundwater conditions outside the potential effects of cemetery leachate; samples from this well were collected near the water table. Nitrogen and phosphorus compounds were present at higher concentrations in two of the three monitoring wells (wells C1 and C3) than in the Fenner well. Formaldehyde and pharmaceuticals were not detected in any of the wells; however, several trace metals, including arsenic, manganese, and aluminum, were present in high concentrations, with arsenic concentrations typically exceeding the U.S. Environmental Protection Agency (EPA) drinking-water standard. Several wastewater indicator compounds, including atrazine, phenol, p-cresol, camphor, and skatole, were detected in the monitoring wells. Microbial data indicate the presence of staphylococci, enterococci, and Escherichia coli (E. coli) , with the highest concentrations being measured in the same two monitoring wells that exhibited elevated concentrations of nutrients in the groundwater (wells C1 and C3). Several bacterial pathogen genes were detected, including several Enterococcus species (spp.)— vanB (vancomycin-resistant enterococci), shiga-toxin-producing E. coli genes (including eaeA [attachment virulence trait] and stx1 [moderate toxin]), and the E. coli 16s ribosomal RNA (rDNA) gene ( E. coli species marker). These results were similar to results of studies conducted in Canada, Australia, and the United Kingdom, in which concentrations of bacteria, metals, and nutrients were elevated in groundwater near cemeteries.

Michigan

Quality of water in the upper Ohio River basin and at Erie, Pennsylvania

This paper discusses the quality of water on the most important tributaries of Ohio River in Pennsylvania, New York, West Virginia, and Maryland, and the nature of the water supply at Erie, Pa. The amount and character of the pollution is described and the results of drinking contaminated water as shown by typhoid statistics are indicated. The conditions on the tributaries of Ohio River in Ohio are discussed in Water-Supply and Irrigation Paper No. 79, United States Geological Survey, pages 129-187. The water supplies and sewerage of small towns high up toward the head of a large drainage system do not in many cases receive the attention they should. Epidemics of a waterborne disease which affect large municipalities near the mouth of the river and therefore attract attention must necessarily have their origin in the pollution of the watershed above. It is evident, therefore, that adequate sanitation of the small towns and a water supply as carefully guarded as that of a large city would prevent disease at its very source and be far less expensive than the costly battles which are waged against epidemics in huge centers of population after disease has broken out. Typhoid fever statistics for small towns in this section are seldom available and are more or less unreliable at best. The few figures given show the existence of virulent typhoid fever in most towns of the drainage areas in certain years, and as these towns drain into the streams the liability ofthe water to infection is evident. The significance of typhoid fever death rates will be better understood from the statistics presented below, which have been collated from a number of cities having excellent water supplies.

Water Supply Paper

Spray vaccination: A method for the immunization of fish

An economical, efficacious vaccine delivery system for immunizing fish has been developed which employs a liquid spray apparatus operated at pressures up to 7.0 kg/cm 2 (0 to 100 lb/in 2 ). A bacterin consisting of formalin-killed Vibrio anguillarum culture was both antigenic and immunogenic when sprayed on coho salmon(Oncorhynchus kisutch)and rainbow trout(Salmo gairdneri). The technique, referred to as spray(shower)vaccination, was found to confer higher levels of immunity against virulent V. anguillarum than oral vaccination.

Fish Pathology

Bat flight and zoonotic viruses

Bats are sources of high viral diversity and high-profile zoonotic viruses worldwide. Although apparently not pathogenic in their reservoir hosts, some viruses from bats severely affect other mammals, including humans. Examples include severe acute respiratory syndrome coronaviruses, Ebola and Marburg viruses, and Nipah and Hendra viruses. Factors underlying high viral diversity in bats are the subject of speculation. We hypothesize that flight, a factor common to all bats but to no other mammals, provides an intensive selective force for coexistence with viral parasites through a daily cycle that elevates metabolism and body temperature analogous to the febrile response in other mammals. On an evolutionary scale, this host–virus interaction might have resulted in the large diversity of zoonotic viruses in bats, possibly through bat viruses adapting to be more tolerant of the fever response and less virulent to their natural hosts.

Emerging Infectious Diseases

Neutralizing monoclonal antibodies recognize antigenic variants among isolates of infectious hematopoietic necrosis virus

eutralizing monoclonal antibodies were developed against strains of infectious hematopoietic necrosis virus (IHNV) from steelhead trout Salmo gairdneri in the Deschutes River of Oregon, chinook salmon Oncorhynchus tshawytscha in the Sacramento River of California, and rainbow trout Salmo gairdneri reared in the Hagerman Valley of Idaho, USA. These antibodies were tested for neutralization of 12 IHNV isolates obtained from salmonids in Japan, Alaska, Washington, Oregon, California, and Idaho. The antibodies recognized antigenic variants among the isolates and could be used to separate the viruses into 4 groups. The members of each group tended to be related by geographic area rather than by source host species, virulence, or date of isolation.

Diseases of Aquatic Organisms

Emergence of MD type infectious hematopoietic necrosis virus in Washington State coastal steelhead trout

Infectious hematopoietic necrosis virus (IHNV) occurs in North America as 3 major phylogenetic groups designated U, M, and L. In coastal Washington State, IHNV has historically consisted of U genogroup viruses found predominantly in sockeye salmon Oncorhynchus nerka . M genogroup IHNV, which has host-specific virulence for rainbow and steelhead trout O. mykiss , was detected only once in coastal Washington prior to 2007, in an epidemic among juvenile steelhead trout in 1997. Beginning in 2007 and continuing through 2011, there were 8 IHNV epidemics in juvenile steelhead trout, involving 7 different fish culture facilities in 4 separate watersheds. During the same time period, IHNV was also detected in asymptomatic adult steelhead trout from 6 coastal watersheds. Genetic typing of 283 recent virus isolates from coastal Washington revealed that the great majority were in the M genogroup of IHNV and that there were 2 distinct waves of viral emergence between the years 2007 and 2011. IHNV type mG110M was dominant in coastal steelhead trout during 2007 to 2009, and type mG139M was dominant between 2010 and 2011. Phylogenetic analysis of viral isolates indicated that all coastal M genogroup viruses detected in 1997 and 2007 to 2011 were part of the MD subgroup and that several novel genetic variants related to the dominant types arose in the coastal sites. Comparison of spatial and temporal incidence of coastal MD viruses with that of the rest of the Pacific Northwest indicated that the likely source of the emergent viruses was Columbia River Basin steelhead trout.

Washington

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