Morphological and molecular characterisation of Aporocotyle margolisi Smith, 1967 (Digenea: Aporocotylidae) from the North Pacific hake Merluccius productus (Ayres) (Gadiformes: Merlucciidae) off Oregon, USA.

Aporocotylid blood flukes conspecific with Aporocotyle margolisi Smith, 1967 were collected from the bulbus arteriosus of the North Pacific hake Merluccius productus (Ayres). This study revisits the morphology of A. margolisi, including drawings, measurements and scanning electron microscopy images, and provides for the first time molecular data for the large subunit of the ribosomal RNA (28S rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) genes for this species. A 28S rDNA phylogenetic study of A. margolisi, and all available Aporocotyle spp., was also performed. The distribution range of A. margolisi is extended to the Pacific coast of the USA. We provide a morphological comparison of Aporocotyle spp. from the Pacific coast in North America as well as other Aporocotyle spp. infecting hake. Comparisons with the original description revealed that the new specimens of A. margolisi were considerably larger with respect to all morphological features, except for shorter spines. Molecular results showed a close relationship between A. margolisi and A. argentinensis Smith, 1969 from the Argentine hake Merluccius hubbsi Marini. The phylogenetic relationships of Aporocotyle spp. point to a possible co-speciation of hakes species and these blood fluke parasites.

Parasites as biological tags of marine, freshwater and anadromous fishes in North America from the Tropics to the Arctic.

Parasites have been considered as natural biological tags of marine fish populations in North America for almost 75 years. In the Northwest Atlantic, the most studied species include Atlantic cod (Gadus morhua), Atlantic herring (Clupea harengus) and the redfishes (Sebastes spp.). In the North Pacific, research has centred primarily on salmonids (Oncorhynchus spp.). However, parasites have been applied as tags for numerous other pelagic and demersal species on both the Atlantic and Pacific coasts. Relatively few studies have been undertaken in the Arctic, and these were designed to discriminate anadromous and resident salmonids (Salvelinus spp.). Although rarely applied in fresh waters, parasites have been used to delineate certain fish stocks within the Great Lakes-St Lawrence River basin. Anisakid nematodes and the copepod Sphyrion lumpi frequently prove useful indicators in the Northwest Atlantic, while myxozoan parasites prove very effective on the coast and open seas of the Pacific Ocean. Relative differences in the ability of parasites to discriminate between fish stocks on the Pacific and Atlantic coasts may be due to oceanographic and bathymetric differences between regions. Molecular techniques used to differentiate populations and species of parasites show promise in future applications in the field.

Widespread and increasing near-bottom hypoxia in the coastal ocean off the United States Pacific Northwest.

The 2021 summer upwelling season off the United States Pacific Northwest coast was unusually strong leading to widespread near-bottom, low-oxygen waters. During summer 2021, an unprecedented number of ship- and underwater glider-based measurements of dissolved oxygen were made in this region. Near-bottom hypoxia, that is dissolved oxygen less than 61 µmol kg-1 and harmful to marine animals, was observed over nearly half of the continental shelf inshore of the 200-m isobath, covering 15,500 square kilometers. A mid-shelf ribbon with near-bottom, dissolved oxygen less than 50 µmol kg-1 extended for 450 km off north-central Oregon and Washington. Spatial patterns in near-bottom oxygen are related to the continental shelf width and other features of the region. Maps of near-bottom oxygen since 1950 show a consistent trend toward lower oxygen levels over time. The fraction of near-bottom water inshore of the 200-m isobath that is hypoxic on average during the summer upwelling season increases over time from nearly absent (2%) in 1950-1980, to 24% in 2009-2018, compared with 56% during the anomalously strong upwelling conditions in 2021. Widespread and increasing near-bottom hypoxia is consistent with increased upwelling-favorable wind forcing under climate change.

Comparison and evaluation of Renibacterium salmoninarum quantitative PCR diagnostic assays using field samples of Chinook and coho salmon.

Renibacterium salmoninarum is a Gram-positive bacterium causing bacterial kidney disease (BKD) in susceptible salmonid fishes. Several quantitative PCR (qPCR) assays to measure R. salmoninarum infection intensity have been reported, but comparison and evaluation of these assays has been limited. Here, we compared 3 qPCR primer/probe sets for detection of R. salmoninarum in field samples of naturally exposed Chinook and coho salmon first identified as positive by nested PCR (nPCR). Additional samples from a hatchery population of Chinook salmon with BKD were included to serve as strong positive controls. The 3 qPCR assays targeted either the multiple copy major soluble antigen (msa) genes or the single copy abc gene. The msa/non-fluorescent quencher (NFQ) assay amplified R. salmoninarum DNA in 53.2% of the nPCR positive samples, whereas the abc/NFQ assay amplified 21.8% of the samples and the abc/TAMRA assay 18.2%. The enzyme-linked immunosorbent assay (ELISA) successfully quantified only 16.4% of the nPCR positive samples. Although the msa/NFQ assay amplified a greater proportion of nPCR positive samples, the abc/NFQ assay better amplified those samples with medium and high ELISA values. A comparison of the geometric mean quantity ratios highlighted limitations of the assays, and the abc/NFQ assay strongly amplified some samples that were negative in other tests, in contrast to its performance among the sample group as a whole. These data demonstrate that both the msa/NFQ and abc/NFQ qPCR assays are specific and effective at higher infection levels and outperform the ELISA. However, most pathogen studies will continue to require multiple assays to both detect and quantify R. salmoninarum infection.

Effect of Nanophyetus salmincola and Bacterial Co-Infection on Mortality of Juvenile Chinook Salmon.

The freshwater trematode Nanophyetus salmincola has been demonstrated to impair salmonid immune function and resistance to the marine pathogen Vibrio anguillarum, potentially resulting in ocean mortality. We examined whether infection by the parasite N. salmincola similarly increases mortality of juvenile Chinook Salmon Oncorhynchus tshawytscha when they are exposed to the freshwater pathogens Flavobacterium columnare or Aeromonas salmonicida, two bacteria that juvenile salmonids might encounter during their migration to the marine environment. We used a two-part experimental design where juvenile Chinook Salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., and then challenged with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare infection was higher in N. salmincola-parasitized fish than in nonparasitized fish. In contrast, cumulative percent mortality from A. salmonicida infection did not differ between N. salmincola-parasitized and nonparasitized groups. No mortalities were observed in the N. salmincola-parasitized-only and control groups from either challenge. Our study demonstrates that a relatively high mean intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae can alter the outcomes of bacterial infection in juvenile Chinook Salmon, which might have implications for disease in wild fish populations.

Temporal changes in the prevalence of parasites in two Oregon estuary-dwelling fishes.

The parasite faunas of juvenile English sole (Parophrys vetulus) in 1971-1972 and staghorn sculpin (Leptocottus armatus) in 1971 from Yaquina Bay, Oregon, were compared with faunas found in the same estuary in 1997-2000 (English sole) and 1999-2000 (staghorn sculpin). The 7 most commonly occurring parasites in 1971 were compared with the same species observed during the same month and sampling sites in 1997-2000. Multivariate community analysis of juvenile English sole parasites supported the suggestion that the 1971 parasite data were representative of the early-1970s time period. Four of the parasite species infecting English sole and 6 of those infecting staghorn sculpins had significantly lower prevalences in 1997-2000. Parasite species with significantly lower prevalences also had reduced intensity levels. One parasite (Glugea stephani) of English sole increased in prevalence in the 1997-2000 samples in association with the warm estuarine temperatures during the 1997 El Niño year. Although the causes for the changes in occurrence of other parasites were not determined, ecological changes in Yaquina Bay that may have influenced parasite ecology include apparent changes in the estuary ichthyofauna that occurred between the sampling periods. Such changes could be associated with increases in the number of California sea lions (Zalophus californianus) subsequent to establishment of the Marine Mammal Protection Act in 1972.

Recent increase in Nybelinia surmenicola prevalence and intensity in Pacific hake (Merluccius productus) off the United States west coast.

A larval marine cestode was found in 82.0% of 834 Pacific hake (Merluccius productus) stomachs collected from 341 trawl stations along the United States west coast during the summers of 2008 and 2009. Morphology and DNA sequencing was used to identify the cestode as Nybelinia surmenicola. In an examination of 131 Pacific hake stomachs collected from the same region in 1999, N. surmenicola prevalence was 35.1%. The results from a general linear model suggested that their prevalence is influenced by year and latitude, Pacific hake size, and sex. Mean intensity of N. surmenicola in 2008-2009 was 20.22 (±1.13 SE) and was positively related to Pacific hake length and the latitude of collection. Year-1 Pacific hake (<27 cm length) had significantly lower prevalence and intensity of N. surmenicola compared to older and larger fish. Pacific hake collected south of Point Conception, California (32.5 to 35°N) had lower prevalence and intensity of N. surmenicola compared to those collected in northern latitudes (35.1 to 48.4°N). Higher N. surmenicola prevalence in Pacific hake in recent years suggests food-web fluctuations in the northern California current ecosystem caused by changes in ocean transport of zooplankton or pelagic fish distributions and warrants future monitoring as a metric for ecosystem change.

Population structure of three species of Anisakis nematodes recovered from Pacific sardines (Sardinops sagax) distributed throughout the California Current system.

Members of the Anisakidae are known to infect over 200 pelagic fish species and have been frequently used as biological tags to identify fish populations. Despite information on the global distribution of Anisakis species, there is little information on the genetic diversity and population structure of this genus, which could be useful in assessing the stock structure of their fish hosts. From 2005 through 2008, 148 larval anisakids were recovered from Pacific sardine (Sardinops sagax) in the California Current upwelling zone and were genetically sequenced. Sardines were captured off Vancouver Island, British Columbia in the north to San Diego, California in the south. Three species, Anisakis pegreffii, Anisakis simplex 'C', and Anisakis simplex s.s., were identified with the use of sequences from the internal transcribed spacers (ITS1 and ITS2) and the 5.8s subunit of the nuclear ribosomal DNA. The degree of nematode population structure was assessed with the use of the cytochrome c oxidase 2 (cox2) mitochondrial DNA gene. All 3 Anisakis species were distributed throughout the study region from 32°N to 50°N latitude. There was no association between sardine length and either nematode infection intensity or Anisakis species recovered. Larval Anisakis species and mitochondrial haplotype distributions from both parsimony networks and analyses of molecular variance revealed a panmictic distribution of these parasites, which infect sardines throughout the California Current ecosystem. Panmictic distribution of the larval Anisakis spp. populations may be a result of the presumed migratory pathways of the intermediate host (the Pacific sardine), moving into the northern portion of the California Current in summer and returning to the southern portion to overwinter and spawn in spring. However, the wider geographic range of paratenic (large piscine predators), and final hosts (cetaceans) can also explain the observed distribution pattern. As a result, the recovery of 3 Anisakis species and a panmictic distribution of their haplotypes could not be used to confirm or deny the presence of population subdivision of Pacific sardines in the California Current system.

Mortality of coho salmon (Oncorhynchus kisutch) associated with burdens of multiple parasite species.

Multiple analytical techniques were used to evaluate the impact of multiple parasite species on the mortality of threatened juvenile coho salmon (Oncorhynchus kisutch) from the West Fork Smith River, Oregon, USA. We also proposed a novel parsimonious mathematical representation of macroparasite distribution, congestion rate, which (i) is easier to use than traditional models, and (ii) is based on Malthusian parameters rather than probability theory. Heavy infections of Myxobolus insidiosus (Myxozoa) and metacercariae of Nanophyetus salmincola and Apophallus sp. occurred in parr (subyearlings) from the lower mainstem of this river collected in 2007 and 2008. Smolts (yearlings) collected in 2007-2010 always harboured fewer Apophallus sp. with host mortality recognised as a function of intensity for this parasite. Mean intensity of Apophallus sp. in lower mainstem parr was 753 per fish in 2007 and 856 per fish in 2008, while parr from the tributaries had a mean of only 37 or 13 parasites per fish, respectively. Mean intensity of this parasite in smolts ranged between 47 and 251 parasites per fish. Over-dispersion (variance to mean ratios) of Apophallus sp. was always lower in smolts compared with all parr combined or lower mainstem parr. Retrospective analysis based on smolt data using both the traditional negative binomial truncation technique and our proposed congestion rate model showed identical results. The estimated threshold level for mortality involving Apophallus sp. was at 400-500 parasites per fish using both analytical methods. Unique to this study, we documented the actual existence of these heavy infections prior to the predicted mortality. Most of the lower mainstem parr (approximately 75%) had infections above this level. Heavy infections of Apophallus sp. metacercariae may be an important contributing factor to the high over-wintering mortality previously reported for these fish that grow and develop in this section of the river. Analyses using the same methods for M.insidiosus and N. salmincola generally pointed to minimal parasite-associated mortality.