Candidate genes for monitoring hydrogen peroxide resistance in the salmon louse, Lepeophtheirus salmonis.

BACKGROUND: Hydrogen peroxide (H2O2) is one of the delousing agents used to control sea lice infestations in salmonid aquaculture. However, some Lepeophtheirus salmonis populations have developed resistance towards H2O2. An increased gene expression and activity of catalase, an enzyme that breaks down H2O2, have been detected in resistant lice, being therefore introduced as a resistance marker in the salmon industry. In the present study the aim was to validate the use of catalase expression as a marker and to identify new candidate genes as additional markers to catalase, related to H2O2 resistance in L. salmonis. METHODS: A sensitive and an H2O2 resistant laboratory strain (P0 generation, not exposed to H2O2 for several years) were batch crossed to generate a cohort with a wide range of H2O2 sensitivities (F2 generation). F2 adult females were then exposed to H2O2 to separate sensitive and resistant individuals. Those F2 lice, the P0 lice and field-collected resistant lice (exposed to H2O2 in the field) were used in an RNA sequencing study. RESULTS: Catalase was upregulated in resistant lice exposed to H2O2 compared to sensitive lice. This was, however, not the case for unexposed resistant P0 lice. Several other genes were found differentially expressed between sensitive and resistant lice, but most of them seemed to be related to H2O2 exposure. However, five genes were consistently up- or downregulated in the resistant lice independent of exposure history. The upregulated genes were: one gene in the DNA polymerase family, one gene encoding a Nesprin-like protein and an unannotated gene encoding a small protein. The downregulated genes encoded endoplasmic reticulum resident protein 29 and an aquaporin (Glp1_v2). CONCLUSIONS: Catalase expression seems to be induced by H2O2 exposure, since it was not upregulated in unexposed resistant lice. This may pose a challenge for its use as a resistance marker. The five new genes associated with resistance are put forward as complementary candidate genes. The most promising was Glp1_v2, an aquaglyceroporin that may serve as a passing channel for H2O2. Lower channel number can reduce the influx or distribution of H2O2 in the salmon louse, being directly involved in the resistance mechanism.

In vitro effect of garlic extract and metronidazole against Neoparamoeba pemaquidensis, page 1987 and isolated amoebae from Atlantic salmon.

Neoparamoeba pemaquidensis believed to be the most prevalent parasite of Atlantic salmon industry in Australia. In the present study, the in vitro effects of crude extract of garlic and metronidazole were investigated using a primary culture toxicity assay. Garlic extract appeared to be completely effective at killing a cultured strain (NP251002) of Neoparamoeba pemaquidensis in vitro at a dilution of 1:100 with in 24 h. The number of viable Amoebae after using garlic extract in lower dilutions (1:200, 1:400, 1:800, 1:1000) for 24 h, also were significantly lower than in the control group. Garlic extract was also efficacious at killing wild type Amoebae that isolated from the diseased fish showing clinical signs of AGD. Metronidazole had no clear effect against Neoparamoeba pemaquidensis (NP251002) even in a concentration of 50 mg L(-1) for 24 h. However some morphological changes have occurred in metronidazole-treated Amoebae after 5 days of exposure.

Relationship between dose of emamectin benzoate and molting response of ovigerous American lobsters (Homarus americanus).

A widely-prescribed treatment to control sea lice on cultured salmon is the administration of feed medicated with SLICE (active ingredient emamectin benzoate (EMB)). High doses of EMB can disrupt the molt cycle of ovigerous American lobsters, causing them to enter proecdysis prematurely and lose their attached eggs when the shell is cast. To determine the dose response to EMB, lobsters were forced to ingest doses that ranged from 0.05 to 0.39 microg g(-1). A significant proportion of lobsters given doses of 0.39 and 0.22 microg g(-1) (37% and 23%, respectively) molted prematurely, almost a year earlier than the control group. All the lobsters in the 0.05 and 0.12 microg g(-1) groups molted at the normal time and the mean time of molt was similar to that of the control group. Thus, the no-observed-effect level (NOEL) and lowest-observed-effect level (LOEL) of EMB on the molt cycle were 0.12 and 0.22 microg EMB g(-1) lobster, respectively. To acquire the LOEL, a 500-g lobster would have to consume 22 g of salmon feed medicated with SLICE at a level of 5 microg EMB g(-1) feed.

Efficacy of the fumagillin analog TNP-470 for Nucleospora salmonis and Loma salmonae infections in chinook salmon Oncorhynchus tshawytscha.

Oral treatment with fumagillin is effective for controlling various microsporean and myxosporean infections in fish. We tested a synthetic analog of fumagillin, TNP-470 (Takeda Chemical Industries), for its efficacy against 2 microsporean pathogens of salmon: Loma salmonae and Nucleospora salmonis. Chinook salmon Oncorhynchus tshawytscha were experimentally infected with either L. salmonae (per os) or N. salmonis (intraperitoneal, i.p., injection) and held in fresh water at 15 degrees C. Fish were then divided into 3 replicate groups: untreated or treated orally at 1.0 mg or at 0.1 mg drug kg-1 fish d-1. With L. salmonae, the high dose fish had 0.32 xenomas mm-2 of gill tissue compared to controls at 24.5 xenomas per mm2. With N. salmonis infections, untreated fish exhibited 100% infection, showed prominent clinical signs (e.g. renal swelling, anaemia), and high mortality. In contrast, fish treated at 1.0 mg kg-1 showed no clinical signs, and 16% of those treated at 0.1 mg kg-1 showed only mild gross pathological changes. With the treated groups, over 50% of the fish exhibited extremely light infections, even with high dose treatments, but no mortalities were attributed to N. salmonis infections. Uninfected fish treated at 1.0 mg drug kg-1 fish d-1 for 5 wk appeared clinically normal and showed no reduction in growth. However, about half of these fish exhibited atrophy of the renal interstitial hematopoietic tissue.