BACKGROUND:The salmon louse, Lepeophtheirus salmonis, is an ectoparasitic copepod which feeds on the mucus, skin and blood of salmonid fish species. The parasite can persist on the surface of the fish without any effective control being exerted by the host immune system. Other ectoparasitic invertebrates produce compounds in their saliva, excretions and/or secretions which modulate the host immune responses allowing them to remain on or in the host during development. Similarly, compounds are produced in secretions of L. salmonis which are thought to be responsible for immunomodulation of the host responses as well as other aspects of crucial host-parasite interactions.METHODS:In this study we have identified and characterised the proteins in the excretory/secretory (E/S) products of L. salmonis using LC-ESI-MS/MS.RESULTS:In total 187 individual proteins were identified in the E/S collected from adult lice and pre-adult sea lice. Fifty-three proteins, including 13 serine-type endopeptidases, 1 peroxidase and 5 vitellogenin-like proteins were common to both adult and pre-adult E/S products. One hundred and seven proteins were identified in the adult E/S but not in the pre-adult E/S and these included serine and cysteine-type endopeptidases, vitellogenins, sphingomyelinase and calreticulin. A total of 27 proteins were identified in pre-adult E/S products but not in adult E/S.CONCLUSIONS:The assigned functions of these E/S products and the potential roles they play in host-parasite interaction is discussed.

Parasitic infections by the salmon louse, Lepeophtheirus salmonis (Krøyer), cause huge economic damage in salmon farming in the northern hemisphere, with combined treatment costs and production losses in 2014 having been estimated at US$ 350 million for Norway (annual production 1.25 million tonnes). The control of L. salmonis relies significantly on medicinal treatments, supplemented by non-pharmacological approaches. However, efficacy losses have been reported for several delousing agents, including the pyrethroid deltamethrin. The aim of the present study was to analyse the genetic basis of deltamethrin resistance in L. salmonis. Deltamethrin median effective concentrations (EC50) were 0.28 μg L-1 in the drug susceptible L. salmonis strain IoA-00 and 40.1 μg L-1 in the pyrethroid resistant strain IoA-02. IoA-00 and IoA-02 were crossed to produce families spanning one parental and three filial generations (P0, F1-F3). In three families derived from P0 crosses between an IoA-00 sire and an IoA-02 dam, 98.8% of F2 parasites (n = 173) were resistant, i.e. remained unaffected after exposure to 2.0 μg L-1 deltamethrin. F3 parasites from these crosses showed a deltamethrin EC50 of 9.66 μg L-1. In two families of the inverse orientation at P0 (IoA-02 sire x IoA-00 dam), 16.7% of F2 parasites were resistant (n = 84), while the deltamethrin EC50 in F3 animals was 0.26 μg L-1. The results revealed a predominantly maternal inheritance of deltamethrin resistance. The 15,947-nt mitochondrial genome was sequenced and compared among six unrelated L. salmonis strains and parasites sampled from wild salmon in 2010. IoA-02 and three further deltamethrin resistant strains, established from isolates originating from different regions of Scotland, showed almost identical mitochondrial haplotypes. In contrast, the mitochondrial genome was variable among susceptible strains and L. salmonis from wild hosts. Deltamethrin caused toxicity and depletion of whole body ATP levels in IoA-00 but not IoA-02 parasites. The maternal inheritance of deltamethrin resistance and its association with mitochondrial haplotypes suggests that pyrethroid toxicity in L. salmonis may involve molecular targets encoded by mitochondrial genes.

Few studies have focussed on the health and immunity of triploid Atlantic salmon and therefore much is still unknown about their response to commercially significant pathogens. This is important if triploid stocks are to be considered for full-scale commercial production. This study aimed to investigate and compare the response of triploid and diploid Atlantic salmon to an experimental challenge with Neoparamoeba perurans, causative agent of amoebic gill disease (AGD). This disease is economically significant for the aquaculture industry. The results indicated that ploidy had no significant effect on gross gill score or gill filaments affected, while infection and time had significant effects. Ploidy, infection and time did not affect complement or anti-protease activities. Ploidy had a significant effect on lysozyme activity at 21 days post-infection (while infection and time did not), although activity was within the ranges previously recorded for sal-monids. Stock did not significantly affect any of the parameters measured. Based on the study results, it can be suggested that ploidy does not affect the manifestation or severity of AGD pathology or the serum innate immune response. Additionally, the serum immune response of diploid and triploid Atlantic salmon may not be significantly affected by amoebic gill disease.

Barley protein concentrate (BPC) was tested as a protein source in the diets of Atlantic salmon post-smolts. Fish were fed one of four experimental diets consisting of a fish meal/soya protein concentrate control (CT) along with two feeds supplemented with increasing levels of BPC replacing the more costly SPC. A fourth diet partially replaced FM in the high BPC diet with a liquid fish protein concentrate (FPC) made from fish trimmings. No significant differences were observed in terms of growth at the end of the 12-week feeding period, although the protein efficiency ratio (g gain g−1 protein consumed) was significantly lower for the control compared to fish fed diets containing either BPC only or FPC and BPC. This suggests that diets containing BPC had a beneficial effect when compared to the control diet. Furthermore, the lack of any detriment to fish growth in diets containing BPC suggests there are no significant issues regarding any negative effects of potential antinutritional factors which can otherwise be the case with other plant origin products. The data presented in this study indicate that BPC and FPC are products which could be of benefit to salmon culture, and related species, in providing a valuable new raw material to the industry.

Dissemination Statement This publication may be re-used free of charge in any format or medium. It may only be reused accurately and not in a misleading context. For material must be acknowledged as SARF copyright and use of it must give the title of the source publication. Where third party copyright material has been identified, further use of that material requires permission from the copyright holders concerned. Disclaimer The opinions expressed in this report do not necessarily reflect the views of SARF and SARF is not liable for the accuracy of the information provided or responsible for any use of the content. Title: Evaluation of sensitivity to chemotherapeutants in successive generations of Lepeoptheirus salmonis from a resistant population.

This study investigated the effect on egg quality of dietary supplementation of Atlantic cod broodstock with the carotenoid astaxanthin (ASTA). Duplicate groups of farm-reared Atlantic cod broodstock were fed either a control diet with no added ASTA, or an ASTA supplemented diet (73.7 mg/kg dry weight; Carophyll Pink (R)) for 2 months prior to peak spawning. The results indicated that ASTA uptake into eggs from the broodstock diet was highly efficient. Fish fed the diet supplemented with ASTA produced fewer batches of eggs, but the mean number per batch of eggs spawned/kg female was higher, and numbers of floating eggs and numbers of fertilised eggs per kg female in each batch were also significantly improved. A correlation between the egg ASTA content and fertilisation success of individual batches was identified. This improvement in egg quality demonstrated the potential value of ASTA supplementation of broodstock diets for cod. ASTA supplementation produced a 20% increase in the number of eggs per batch spawned, a 37% increase in the number per batch of floating eggs per kg female and a 47% increase in the number per batch of fertilised eggs per kg female. These results clearly demonstrate significant benefits of ASTA supplementation of cod broodstock feeds in terms of improved egg quality and larval production. (C) 2008 Elsevier B.V. All rights reserved.

The depletion of emamectin B-1a in the edible tissues of rainbow trout (Oncorhynchus mykiss) was studied at two temperatures following treatment with emarnectin benzoate (Slice((R))) in feed. Fish approaching market size (400-1500 g) were held in tanks supplied with temperature-controlled seawater at 6 +/- 1 degrees C (cold water) or 15 +/- 1 degrees C (warm water). In each study the medicated group was offered feed containing emamectin benzoate at a nominal dose rate of 50 mu g kg(-1) fish day(-1) for 7 days and the control group was offered unmedicated feed. Actual dose rates, calculated from growth rate and feed consumption data, and measured emamectin benzoate concentrations in feed, were 88.6% nominal in the cold water study (96.6% adjusted for feed assay recovery) and 96.8% nominal in the warm water study (105.1% adjusted for feed assay recovery).Concentrations of emamectin B-1a were determined in fillet samples collected at intervals from 6 h to 77 days post-treatment in the cold water study and 6 h to 49 days post-treatment in the warm water study. In the cold water study, mean emamectin B-1a residues ranged from 81.8 +/- 44.5 ng g(-1) at 1 day post-treatment (102.3 +/- 55.7 ng g(-1) adjusted for recovery) to 13.7 +/- 10.5 ng g(-1) at 77 days post-treatment (17.2 +/- 13.1 ng g(-1)). In the warm water study, mean residue concentrations ranged from 64.5 +/- 50.3 ng g(-1) at 6 h post-treatment (80.7 +/- 62.9 ng g(-1) adjusted for recovery) to 1.6 +/- 1.6 ng g(-1) at 49 days post-treatment (2.0 +/- 2.0 ng g(-1)). In the cold water study, residues in skin and muscle were also determined separately. On average, emarnectin Bla concentrations in skin were approximately 1.8 times higher than in muscle.Measured residue levels ranged widely and no detectable residues were found in at least a few individual fish at all time points. This high variability was considered to be due to differences in medicated feed consumption within the experimental population. Depletion of emamectin was faster at 15 degrees C than at 6 degrees C. In both studies the depletion curve showed a small secondary peak at around 90 degree-days. This observation is consistent with recirculation of the compound from a body store. (c) 2006 Elsevier B.V. All rights reserved.

In recent years, interest in the intensive culture of Atlantic cod (Gadus morhua) has increased dramatically due to several factors including a reduced supply of cod from capture fisheries, a high market price for wild cod and the suitability of the species for culture. One of the major problems facing the industry has been the high cost and unreliability of live feeds, specifically the live feed Artemia. The main objective of this project was to determine whether Artemia use could be reduced or replaced completely with two novel microparticulate diets (MPD's), without negatively compromising growth, survival performance and tank hygiene under simulated commercial conditions.The experiment consisted of four treatments, a live feed control treatment (group A), a 50% Artemia replacement treatment with MPD-1 (group B), a 100% Artemia replacement treatment with MPD-1 (group C) and a 100% Artemia replacement treatment with MPD-2 (group D). All treatments were run in triplicate. Growth performances, development (standard length, eye diameter, myotome height and wet weight), water quality (bacteriology and spectrophotometry) and survival were measured throughout the duration of the trial.The results of the experiment indicate that the treatments containing Arteinia (groups A and B) both achieved significantly higher growth rates than treatments that did not contain Artemia (groups C and D). The highest survival rates achieved at 70 dph were in treatments A and D (13.8%+/- 0.7% and 14.2%+/- 2.1% respectively) when compared to treatments B and C (11.8%+/- 0.3% and 5.5%+/- 1.3% respectively). Survival was also significantly higher in treatment B than in treatment C. This study demonstrates that while the best growth and survival rates are still achieved when cod larvae are fed Artemia, combining live feeds and commercially available MPD's (co-feeding) can produce comparable growth and survival rates thus potentially reducing the reliance on live feeds. However the complete replacement of Artemia with MPD's still significantly reduced growth potential suggesting that the nutritional composition of MPD's, requires further investigation. (c) 2006 Elsevier B.V. All rights reserved.

Lipids and essential fatty acids, particularly the highly unsaturated fatty acids, 20:5n-3 (eicosapentaenoic acid; EPA), 22:6n-3 (docosahexaenoic acid; DHA) and 20:4n-6 (arachidonic acid, AA) have been shown to be crucial determinants of marine fish reproduction directly affecting fecundity, egg quality, hatching success, larval malformation and pigmentation. In Atlantic cod (Gadus morhua L.) culture, eggs from farmed broodstock can have much lower fertilization and hatching rates than eggs from wild broodstock. The present study aimed to test the hypothesis that potential quality and performance differences between eggs from different cod broodstock would be reflected in differences in lipid and fatty acid composition. Thus eggs were obtained from three broodstock, farmed, wild/fed and wild/unfed, and lipid content, lipid class composition, fatty acid composition and pigment content were determined and related to performance parameters including fertilization rate, symmetry of cell division and survival to hatching. Eggs from farmed broodstock showed significantly lower fertilization rates, cell symmetry and survival to hatching rates than eggs from wild broodstock. There were no differences in total lipid content or the proportions of the major lipid classes between eggs from the different broodstock. However, eggs from farmed broodstock were characterized by having significantly lower levels of some quantitatively minor phospholipid classes, particularly phosphatidylinositol. There were no differences between eggs from farmed and wild broodstock in the proportions of saturated, monounsaturated and total polyunsaturated fatty acids. The DHA content was also similar. However, eggs from farmed broodstock had significantly lower levels of AA, and consequently significantly higher EPA/AA ratios than eggs from wild broodstock. Total pigment and astaxanthin levels were significantly higher in eggs from wild broodstock. Therefore, the levels of AA and phosphatidylinositol, the predominant AA-containing lipid class, and egg pigment content were positively related to egg quality or performance parameters such as fertilization and hatching success rates, and cell symmetry.

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

Atlantic cod is a species with a high biological potential for aquaculture. Nowadays, as a result of the decline of natural stocks and increase in market value of cod, it is regarded as an alternative species with high potential for commercial farming. Therefore, one of the priorities for development of a profitable commercial activity is formulation of cost-effective diets.The present work was developed with the aim of testing four extruded diets differing in the levels of protein (48% or 58%) and lipid (12% and 16%). A second objective was to verify whether the producers could profit from the lipid-rich livers characteristic of farmed cod, exploiting it as a source of cod liver oil for human health purposes.A 16-week growth trial was conducted with juvenile cod (233 g, average total weight) using three replicates per treatment. Length measurements were performed every 4 weeks and samples of muscle and liver were taken for total lipid, lipid classes and fatty acid determination, proximate analysis and liver histology. The effect of the diets was evaluated based on growth (SGR and GF3), feeding performance (FCE, PER), condition, hepatosomatic index and biochemical composition.The tested diets induced good performance results, with a protein sparing effect by lipid and a more efficient use of protein in diets with a lower protein content being clear at 12 weeks but not so obvious after 16 weeks of culture. It is the authors' opinion that the 48/16 diet appeared to be the best compromise between growth, feed utilisation and cost. The diets with a higher lipid level induced a higher HSI and lipid content in the liver, The muscle and particularly the liver FA profile, reflected that of the diets. The FA profile of farmed cod liver oil showed promising properties, despite having a slightly higher content of saturated fatty acids and linoleic acid, compared to "natural" cod liver oil. (C) 2001 Elsevier Science B.V. All rights reserved.

E. (1992) Pathological changes in kidneys of dogs with natural Leishmania infection. Veterinary Parasitology 45, 33-47 POLI, A., ABRAMO, F., MANCIANTI, F., NIGRO, M., PIERI, S. & BIONDA, A. (1991) Renal involvement in canine leishmaniasis. Nephronl 57,444-452 PUMAROLA, M., BREVIK, L., BADIOLA, J., VARGAS, A., DOMINGO, M. & FERRER, L. (1991) Canine leishmaniasis associated with systemic vasculitis in two dogs. Journal of Comparative Pathology 105, 279-286 REAGAN, W. J. & REBAR, A. H. (1995) Platelet disorders. In Textbook of Veterinary Internal Medicine. Philadelphia, W. B. Saunders. pp 1964-1976 SAINZ, A., TESOURO, M. A., RODRIGUEZ, F., MAYORAL, I. & MAZZUCHELLI, F. (1995) Seroprevalence of Ehrlichia canis infections in police dogs in Spain. Preventive Veterinary Medicine 23, 179-182 SLAPPENDEL, R. J. (1988) Canine leishmaniasis. A review based on 95 cases in the Netherlands. Veterinary Quarterly 10, 1-16 SLAPPENDEL, R. J. (1989) Disseminated intravascular coagulation. In Kirk's Current Veterinary Therapy X. Small Animal Practice. Philadelphia, W. B. Saunders. pp 451-457 SLAPPENDEL, R. J. & FERRER, L. (1998) Leishmaniasis. In Infectious Diseases of the Dog and Cat. Philadelphia, W. B. Saunders. pp 450-458 SLAPPENDEL, R. J., FRIELINK, R. A., MOL, J. A., NOORDZIJ, A. & HAMER, R. (1992) An enzyme-linked immunosorbent assay (ELISA) for von Willebrand factor antigen (vWf-Ag) in canine plasma. Veterinary Immunology and Immunopathology 33, 145-154 TABOADA, J. & MERCHANT, S. R. (1995) Protozoal and miscellaneous infections. In Textbook of Veterinary Internal Medicine. Philadelphia, W. B. Saunders. pp 384-397 TESOURO, M. A. (1984) Aspectos clinicos y laboratoriales de la leishmaniosis canina. Estudio epizootiologico en la provincia de Madrid. PhD thesis, Universidad Comlutense de Madrid, Madrid TESOURO, M. A., RODRIGUEZ, F., SAINZ, A. & JIMENEZ, F. (1992) Contribuciones al diagnostico y patocronia de la leishmaniosis canina (I). Informacion Veterinaria 125,40-46 VALLADARES, J. E., RUIZ DE GOPEGUI, R., RIERA, C., ALBEROLA, J., GALLEGO, M., ESPADA, Y., PORTUS, M. & ARBOIX, M. (1998) Study of haemostatic disorders in experimentally induced leishmaniasis in Beagle dogs. Research in Veterinary Science 64, 195-198 VARELA, F. (1992) Immunopatologia de la leishmaniosis canina: bases teoricas y aspectos practico. Premios Fundacion Purina, Columna Ediciones SA. pp11 -54 WARDROP, K. J., DHEIN, C. R., FRENIER, S. & MEYERS, K. M. (1989) Altered hemostasis in a dog with chronic renal failure. Journal oftheAmerican Animal Hospital Association 25, 325-329 WERNER, L. L., TURNWALD, G. H. & BARTA, 0. (1994) Immunologic and plasma protein disorders. In Small Animal Clinical Diagnosis by Laboratory Methods. Philadelphia, W. B. Saunders. pp 253-272 WILLIS, S. E., JACKSON, M. L., MERIC, S. M. & ROUSSEAUX, C. G. (1989) Whole blood platelet aggregation in dogs with liver disease. American Journal of Veterinary Research 50, 1893-1897

Atlantic salmon weight range 289-484 g and rainbow trout weight range 166-387 g , held in seawater, were fed medicated diets containing graded levels of emamectin benzoate for 7 days. y1 y1 Nominal dose rates were 0, 100, 250 and 500 mg kg body weight day equivalent to 0, 2, 5 . and 10 times recommended dose rate for treatment of sea lice . Fish were observed for a further 7 days to determine the effects of treatment. Calculated actual dose rates, compensated for feed concentrations determined by chemical analysis, feed intake and weight gain during the course of the study, were 0, 70, 173 and 356 m gk g y1 body weight dayy1 for Atlantic salmon, and 0, 88, 218 and 413 m gk g y1 body weight dayy1 for rainbow trout. No mortality was observed, which could be related to treatment and unequivocal signs of toxicity were recorded only at the highest dose used in these studies. Signs of emamectin benzoate toxicity in both species included lethargy, dark coloration and inappetance. Atlantic salmon also showed loss of coordination. Fish in the high-dose groups exhibiting signs of toxicity showed no evidence of recovery during the 7-day post-treatment period. No pathognomonic signs of emamectin benzoate toxicity were identified

Atlantic salmon (weight range 289–484 g) and rainbow trout (weight range 166–387 g), held in seawater, were fed medicated diets containing graded levels of emamectin benzoate for 7 days. Nominal dose rates were 0, 100, 250 and 500 μg kg−1 body weight day−1 (equivalent to 0, 2, 5 and 10 times recommended dose rate for treatment of sea lice). Fish were observed for a further 7 days to determine the effects of treatment. Calculated actual dose rates, compensated for feed concentrations determined by chemical analysis, feed intake and weight gain during the course of the study, were 0, 70, 173 and 356 μg kg−1 body weight day−1 for Atlantic salmon, and 0, 88, 218 and 413 μg kg−1 body weight day−1 for rainbow trout. No mortality was observed, which could be related to treatment and unequivocal signs of toxicity were recorded only at the highest dose used in these studies. Signs of emamectin benzoate toxicity in both species included lethargy, dark coloration and inappetance. Atlantic salmon also showed loss of coordination. Fish in the high-dose groups exhibiting signs of toxicity showed no evidence of recovery during the 7-day post-treatment period. No pathognomonic signs of emamectin benzoate toxicity were identified during gross necropsy or histopathological examination.

Pseudomyxoma peritonei is a condition characterized by the production of large amounts of mucopolysaccharide by a neoplastic epithelium. Although surgical debulking and removal of the mucinous ascites may be attempted, complete removal of the material is often impossible. Intraperitoneal lavage with 10% dextrose in water (D10W) has been advocated to prevent reaccumulation of the mucus and complications such as bowel obstruction requiring repeat laparotomy. We describe a patient undergoing operation for a large abdominopelvic mass. At laparotomy, a mucinous cystadenocarcinoma of the ovary was found with a great deal of tenacious, mucinous ascites and peritoneal implants. In an effort to more efficiently remove the mucus and prevent subsequent reaccumulation, intraperitoneal irrigation with 10% dextrose in water (D10W) was performed. The patient, who gave no history of prior glucose intolerance, was soon thereafter found to be profoundly hyperglycemic (serum glucose >500 mg/dl). She was treated with insulin and recovered without evident sequelae. Practitioners should be aware of this potentially dangerous complication associated with intraperitoneal dextrose instillation.

The α-inhibin gene has been shown in knockout mouse models to be a suppressor of granulosa tumorigenesis in the mouse. To determine if α-inhibin has the same function in humans, we have assessed the frequency of loss of heterozygosity (LOH) of the α-inhibin gene locus on chromosome 2q in 17 human granulosa cell tumors and 36 epithelial ovarian cancers. LOH was detected in 12 of 36 (33.3%) epithelial tumors but in only 1 of 17 (6%) granulosa cell tumors. These data suggest that in contrast to the suggestions from the mouse model α-inhibin does not function as a granulosa cell tumor suppressor gene in the human. Furthermore, analysis of the TP53 gene in the granulosa cell tumors failed to detect either LOH or point mutations, indicating that they have a developmental pathway distinct from that of epithelial ovarian tumors.

Plasma and liver vitamin E concentrations in a population of farmed Atlantic salmon, Salmo salar L., fed commercial diets were measured by high-performance liquid chromatography. In healthy fish fed diets containing 160-210 mg kg−1all-racα-tocopheryl acetate, vitamin E accumulated in the liver and plasma. Over a 20-month period, beginning 4 months prior to seawater transfer, mean vitamin E concentrations ranged from 51 to 754 μg g−1 wet tissue in liver and from 7 to 68 μg mL−1 in plasma. In liver, a sharp increase in vitamin E concentrations was recorded between 6 and 10 weeks after transfer. In plasma a similar increase occurred between 4 and 5 months post transfer. Total lipid and polyunsaturated fatty acid concentrations were also measured in these tissues. Liver total lipid concentrations fell during the period of smoltification, but there was no relationship between vitamin E and either total lipid or total polyunsaturated fatty acid concentrations in liver or plasma.

Twenty-four general-practice trainees were assessed at the beginning and end of their training year. They improved significantly in their factual recall and problem-solving ability. Sixteen young principals who had had an unstructured training for general practice were assessed at the same time and were initially of the same standard but their results showed no difference over the same year.