Threats to at-risk salmonids of the Canadian Rocky Mountain region

Trout and charr, members of the salmonid family, have high conservation value but are also susceptible to anthropogenic threats in part due to the specificity of their habitat requirements. Understanding historical and future threats facing these species is necessary to promote their recovery. Of freshwater trout and charr in the Canadian Rocky Mountain region, westslope cutthroat trout (Oncorhynchus clarkii lewisi), bull trout (Salvelinus confluentus; a charr species) and Athabasca rainbow trout (Oncorhynchus mykiss) are of conservation concern. And indeed, range contractions and declining populations are evident throughout much of their ranges. Range contraction was most evident in the southern Alberta designatable unit (DU) of westslope cutthroat trout. Diminished populations were also evident in the downstream watersheds of the Alberta bull trout range, and throughout the Athabasca rainbow trout range. We assessed historical and future threats to evaluate the relative importance of individual threats to each DU and compare their impact among species. Individual threats fall into the broad categories of angling, non-native species and genes, habitat loss and alteration, and climate change. Severity of each threat varies by DU and reflects the interaction between species’ biology and the location of the DU. Severity of threats facing each DU has changed over time, reflecting extirpation of native populations, changes in management and industry best practices, expansion of non-native species and progressing climate change. The overall threat impact for each DU indicates a high probability of substantial and continuing declines and calls for immediate action.     

Verrucosamide,a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD₅₀ of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD₅₀ of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.     

Real-time RT-qPCR assay to detect sequences in the Piscine orthoreovirus-1 genome segment S1 associated with heart and skeletal muscle inflammation in Atlantic salmon

During the last decade, Piscine orthoreovirus was identified as the main causative agent of heart and skeletal muscle inflammation (HSMI) in Atlantic Salmon, Norway. A recent study showed that PRV-1 sequences from salmonid collected in North Atlantic Pacific Coast (NAPC) grouped separately from the Norwegian sequences found in Atlantic Salmon diagnosed with HSMI. Currently, the routine assay used to screen for PRV-1 in NAPC water and worldwide cannot differentiate between the two groups of PRV-1. Therefore, this study aimed at developing a real-time polymerase chain reaction (RT-qPCR) assay to target the PRV-1 genome segments specific for variants associated with HSMI. The assay was optimized and tested against 71 tissue samples collected from different regions including Norway, Chile and both coast of Canada and different hosts farmed Atlantic Salmon, wild Coho Salmon and escaped Atlantic Salmon collected in British Columbia, West Coast of Canada. This assay has the potential to be used for screening salmonids and non-salmonids that may carry PRV-1 potentially causing HSMI.     

A comparison of the use of different swab materials for optimal diagnosis of amoebic gill disease (AGD) in Atlantic salmon (Salmo salar L.)

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab^® (calcium alginate fibre-tipped), Isohelix^® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms’ DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.     

Inhibiting fungus on largemouth bass eggs with copper sulfate and its toxicity to fry and juveniles

This study determined the effectiveness of copper sulfate (CuSO₄) to inhibit fungal growth (caused by Saprolegnia spp.) on largemouth bass (LMB) eggs spawned on/in fiber mats in very high-alkalinity/-hardness waters; experiments also determined the toxicity of CuSO₄ to LMB fry and juveniles. An untreated control and three CuSO₄ concentrations (10, 20, and 40 mg/L) were tested under a flow-through scenario in the effectiveness experiment. Eggs were treated daily until hatching began. Fungal load at the time of hatch had a total area > 3.0 cm² in the untreated controls, total area 1.0 to <2.0 cm² in the 10 and 20 mg/L CuSO₄ treatments, and total area > 1.0 cm² in the 40 mg/L CuSO₄ treatments. Fungus samples were identified as Saprolegnia australis. The 24-hr median lethal concentration (LC50) values on the LMB yolk-sac and swim-up fry were 32.0 and 4.6 mg/L CuSO₄, respectively; the No Observed Effect Concentrations (NOEC) were 16.0 and 0.125 mg/L CuSO₄, respectively. Juvenile LMB were extremely tolerant to CuSO₄, and their 24-hr LC50 value was 185.5 mg/L; the NOEC was 64 mg/L. This study indicates that CuSO₄ can be an important resource for hatcheries to control egg fungus, especially in high-alkalinity/-hardness waters.     

Impact of weather variables and season on sporulation of Phytophthora pluvialis and Phytophthora kernoviae

Phytophthora pluvialis and Phytophthora kernoviae are the causal agents of important needle diseases on Pinus radiata in New Zealand. Little is known about the epidemiology of the diseases, making the development of control strategies challenging. To investigate the seasonality and climatic drivers of sporulation, inoculum traps, consisting of pine fascicles floating on water in plastic containers, were exchanged fortnightly at five sites in P. radiata plantations between February 2012 and December 2014. Sections of needle baits were plated onto selective media and growth of Phytophthora pluvialis and P. kernoviae recorded. To explore the generalizability of these data, they were compared to detection data for both pathogens from the New Zealand Forest Health Database (NZFHDB). Further, equivalent analyses on infection of Rhododendron ponticum by P. kernoviae in Cornwall, UK allowed the comparison of the epidemiology of P. kernoviae across different host systems and environments. In New Zealand, inoculum of P. pluvialis and P. kernoviae was detected between January–December and March–November, respectively. Inoculum of both species peaked in abundance in late winter. The probability of detecting P. pluvialis and P. kernoviae was greater at lower temperatures, while the probability of detecting P. pluvialis also increased during periods of wet weather. Similar patterns were observed in NZFHDB data. However, the seasonal pattern of infection by P. kernoviae in the UK was the opposite of that seen for sporulation in New Zealand. Phytophthora kernoviae was likely limited by warmer and drier summers in New Zealand, but by colder winter weather in the UK. These results emphasize the importance of considering both environmental drivers and thresholds in improving our understanding of pathogen epidemiology.     

Strangulation of the duodenum just oral to its caudal flexura caused by a volvulus

An 8-year-old Haflinger mare with a 10-day-old foal at foot was referred with mild abdominal pain. On the basis of clinical examination, blood and peritoneal fluid analysis, the cause of pain was suspected septic peritonitis. The horse was initially managed medically, with the horse placed in box confinement under close monitoring. No permission was given for an exploratory laparotomy, and it was decided to perform a diagnostic laparoscopy in the standing horse. The laparoscopy revealed a 360-degree anticlockwise partial duodenal volvulus of the caudal descending duodenum, which was manually reduced (hand-assisted laparoscopy). After the release, the mesentery of the duodenum showed an ischaemic band (yellowish white and dark red with a hyperaemic side) and the duodenum had a distinct ischaemic ring (orally and aborally) distinguishing the incarcerated part from the normal duodenum. Ten minutes after the release, the colour remained the same. In conjunction with the owner, it was decided to euthanise the pony. Pathology confirmed the presence of a strangulation ring and the ischaemic condition of the involved duodenum.     

Control of insect pests using trunk injection in a newly established apple orchard

We evaluated the potential for using preplant trunk injections of emamectin benzoate in nonbearing apple trees. Trees were evaluated for pest injury and emamectin residues throughout the planting season and into the following year. Injections into the trunk best delivered emamectin benzoate to the canopy compared with injections into the taproot, and the higher rate reduced insect pests more than the lower rate. In the following year, differences in insect control between trunk and root injections were less pronounced, but the higher rate of emamectin benzoate persisted longer and better reduced pests relative to the other treatments.     

Effect of apple cultivars and storage periods on the virulence of Neofabraea spp.

Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.