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Sea lice represent a persistent and growing problem, challenging the resilience and growth of the salmon aquaculture industry. In this Norwegian case study, we studied and discuss how the absence of policy instruments directed at stimulating breeding for lice resistance (LR) might be explained. We found well-documented opportunities for selection progress for LR. Hence, breeding on LR appears with an untapped potential. We discuss how market-based, legal, institutional and interest-based factors can explain the absence of policy instruments stimulating LR breeding. Methodologically, we obtained data from document and literature studies and interviews with key players (salmon breeders, farmers, nongovernmental organizations (NGOs) and governmental bodies in Norway). First, LR is a polygenic trait, which makes it poorly suited for patenting. Furthermore, if only a small proportion of fish farmers choose seeds with higher LR, other operators can easily take on the free-rider role because they will not suffer from reduced gain in growth performance as a result of a much stronger emphasis on LR in the breeding goal. The market is thus not expected to stimulate stronger selection for LR in Norwegian salmon breeding. Second, neither genetic engineering (e.g., gene editing), still struggling with consumer acceptance, nor the uncertainty associated with possible changes in the Norwegian Gene Technology Act stimulate investment in LR via, for example, CRISPR technology. Thirdly, public policy instruments in their entirety have targeted other types of innovations against salmon lice, and none have so far been used to stimulate breeding companies to emphasize LR more strongly in their breeding programmes. From a political point of view, it seems that breeding has been left to the market and the private sector. However, neither the NGOs nor the public seem to be aware of, or pay significant attention to, the breeding potential to improve LR and fish welfare. Fragmented management of the aquaculture sector can camouflage the close ties between political and business interests. The industry is hesitant to invest significantly in long-term breeding targets such as significantly higher genetic LR. This may strengthen the assumption that strong economic interests will reduce the role of science in knowledge-based management. As farmed salmon are increasingly being exposed to stressful delousing treatments, mortality and associated welfare problems have increased significantly. For instance, large fish die more often from cardiomyopathy syndrome (CMS), resulting in growing demand for CMS-resistant salmon. This gives rise to a paradoxical situation: increasing treatments with high mortality and fish welfare issues in farmed salmon, while the lice threat to wild salmon persists., (© 2023 NOFIMA and The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

Background: There has been an ongoing debate in the reproductive field about whether mammalian spermatozoa rely on glycolysis, oxidative phosphorylation or both for their energy production. Recent studies have proposed that human spermatozoa depend mainly on glucose for motility and fertilization but the mechanism behind an efficient glycolysis in human spermatozoa is not well understood. Here, we demonstrate how human spermatozoa utilize exogenous pyruvate to enhance glycolytic ATP production, motility, hyperactivation and capacitation, events that are crucial for male fertility., Methods: Purified human spermatozoa from healthy donors were incubated under capacitating conditions (including albumin, bicarbonate and glucose) and tested for changes in ATP levels, motility, hyperactivation and tyrosine phosphorylation after treatment with pyruvate. The experiments were repeated in the presence of sodium cyanide in order to assess the contribution from mitochondrial respiration. The metabolism of (13)C labeled glucose and pyruvate was traced by a combination of liquid chromatography and mass spectrometry., Results: The treatment of human spermatozoa with exogenous pyruvate increased intracellular ATP levels, progressive motility and hyperactivation by 56, 21 and 130%, respectively. In addition, added pyruvate induced a significant increase in tyrosine phosphorylation levels. Blocking of the electron transport chain did not markedly affect the results, indicating that the mechanism is independent of oxidative phosphorylation. However, the observed effects could be counteracted by oxamate, an inhibitor of lactate dehydrogenase (LDH). Metabolic tracing experiments revealed that the observed rise in ATP concentration resulted from an enhanced glycolytic flux, which was increased by more than 50% in the presence of exogenous pyruvate. Moreover, all consumed (13)C labeled pyruvate added was converted to lactate rather than oxidized in the tricarboxylic acid cycle., Conclusions: Human spermatozoa seem to rely mainly, if not entirely, on glycolysis as the source of ATP fueling the energy-demanding processes of motility and capacitation. The efficient glycolysis is dependent on exogenous pyruvate, which indirectly feeds the accelerated glycolysis with NAD(+) through the LDH-mediated conversion of pyruvate to lactate. Pyruvate is present in the human female reproductive tract at concentrations in accordance with our results. As seen in other mammals, the motility and fertility of human spermatozoa seem to be dictated by the available energy substrates present in the conspecific female.

Background: Post-transcriptional processing of pre-mRNA takes place in several steps and requires involvement of a number of RNA-binding proteins. How pre-mRNA processing is regulated is in large enigmatic. The catalytic (C) subunit of protein kinase A (PKA) is a serine/threonine kinase, which regulates numerous cellular processes including pre-mRNA splicing. Despite that a significant fraction of the C subunit is found in splicing factor compartments in the nucleus, there are no indications of a direct interaction between RNA and PKA. Based on this we speculate if the specificity of the C subunit in regulating pre-mRNA splicing may be mediated indirectly through other nuclear proteins., Results: Using yeast two-hybrid screening with the PKA C subunit Cbeta2 as bait, we identified the G-patch domain and KOW motifs-containing protein (GPKOW), also known as the T54 protein or MOS2 homolog, as an interaction partner for Cbeta2. We demonstrate that GPKOW, which contains one G-patch domain and two KOW motifs, is a nuclear RNA-binding protein conserved between species. GPKOW contains two sites that are phosphorylated by PKA in vitro. By RNA immunoprecipitation and site directed mutagenesis of the PKA phosphorylation sites we revealed that GPKOW binds RNA in vivo in a PKA sensitive fashion., Conclusion: GPKOW is a RNA-binding protein that binds RNA in a PKA regulated fashion. Together with our previous results demonstrating that PKA regulates pre-mRNA splicing, our results suggest that PKA phosphorylation is involved in regulating RNA processing at several steps.

In order to facilitate epidemiological investigations a subdivision of Staphylococcus aureus strains belonging to the 94,96 complex by means of two experimental phages, 16 and 47A, was performed. These phages were selected from the nine experimental phages initially examined because they gave the greatest discrimination. On the basis of reactions with these two phages, 2199 isolates which reacted with phages 94 and 96, and 773 isolates which reacted with phage 96 alone, were each subdivided into two major and two minor groups. Strains with different phage patterns were in a few cases (2/64) isolated from the same deep body site in a patient, and lysogenisation experiments suggested that differences in phage patterns were determined by the presence of prophages. Strains with the phage patterns 94/96 and 96 were found to be unevenly distributed throughout Denmark. This regional distribution suggested that particular strains might predominate in some areas. The extended phage patterns with the experimental phages did not give any retrospectively useful epidemiological information. It is proposed that in future phages 16 and 47A be used for specific investigations into the sources and relatedness of strains involved in small incidents.

484 isolates of P. aeruginosa were obtained from the respiratory tract of 45 out of 70 cystic fibrosis (CF) patients of monthly examinations during one year at the Danish CF Centre. All isolates were serogrouped (O-antigens) and phage typed, and in this way 99 per cent of the isolates could be grouped and/or typed. The isolates from CF patients belonged to many different sero-groups, but 55 per cent were polyagglutinable, and most of these belonged to the 0-3/9 complex. This was significantly different from the distribution into O-groups of isolates obtained from non-CF patients. The results of the combined phage typing and sero-grouping of the isolates revealed the occurrence of 13 clusters of distinct epidemiological types of P. aeruginosa among small groups (2-10 individuals) of CF patients. The predominating endemic strain, which was isolated from 10 (22 per cent) of the patients, belonged to the 0-3/9 complex and was lysed by phage 109, either alone or in combination with a few other phages. Furthermore, in a few cases the eradication of another strain of P. aeruginosa by chemotherapy was followed by colonization of the lungs with the above-mentioned predominating strain, and this strain was associated with high lethality. According to these results, measures should be undertaken to identify and eliminate routes of cross-infection in CF centres in order to diminishe the prevalence of P. aeruginosa infection and thereby reduce the lethality of CF patients.

A total of 6,253 cases of Staphylococcus aureus bacteremia, including 274 (4.4%) endocarditis cases, were registered in Denmark in the period 1975-1984. Patients with hematological malignancies and/or agranulocytosis accounted for 479 of the bacteremia cases. The incidence of endocarditis in this group of patients was only 0.4% as compared to 4.7% in other patients with staphylococcal bacteremia (p less than 0.01). The lower incidence of endocarditis complicating bacteremia in these patients may justify a shorter course of therapy than usually recommended for suspected endocarditis. Patients with hematological malignancies and other patients with agranulocytosis had a higher mortality (49 and 46%, respectively) than other patients with S. aureus bacteremia (33%). The highest mortality was found in patients with multiple myeloma (71%, p less than 0.01), the lowest in patients with acute lymphocytic leukemia (28%, p less than 0.01). The higher mortality in these patients may indicate that empiric antibiotic regimens in granulocytopenic patients should include a specific anti-staphylococcal agent.

In the period 1975-1981, 4060 cases of Staphylococcus aureus bacteremia were recorded in Denmark, and the corresponding strains were examined. The percentage of strains, resistant to penicillin only, rose to 82, and the percentage of multiply-resistant strains fell to five. Newer phage types (94, 96 and 95) increased from 10% to 27% of the material. These strains were usually resistant only to penicillin, but produced large amounts of penicillinase. The ample penicillinase production has also been characteristic for previous epidemic strains, and it is furthermore correlated to mortality. The overall mortality of 34.6% was lower than that of the preceding period. Mortality rates were highest in elderly patients, nosocomial cases, patients with serious primary diseases and endocarditis cases.

During the years 1966-1974, 167,297 strains isolated from 167,297 patients or staff members in Danish hospitals were registered at Staten Seruminstitu. All the strains were phage-typed and examined for production of a 'Tween'-80-splitting enzyme and resistance to mercuric chloride. 158,236 strains were examined for resistance to antibiotics. Since 1968, a steep decrease in the number of strains resistant to three or more antibiotics (multiple-resistant) and in strains of the 83A complex was noticed. In recent years an increase in strains belonging to phage-group I and in those referred to as miscellaneous and non-identified was registered. The increase in the non-typable strains might be explained by the shift of the concentration of the typing phages from 1000 X RTD to 100 X RTD. It is concluded that at least two factors may have contributed to the reduction of the multiple-resistant strains: an altered antibiotic policy, restricting the use of streptomycin and tetracyclines, and an improved hospital hygiene, diminishing the spread of identical strains within the various departments. However, it is emphasized that the consumption of methicillin is still increasing.

In 1979 seven gentamicin (G)-resistant (res.) Staphylococcus aureus strains caused epidemic episodes in eight Danish hospitals. Furthermore, a total of 37 resistant strains were isolated from separate incidents. All the G-res. strains were resistant to kanamycin, sisomicin and tobramycin, five to amikacin and two to cephalothin. None were resistant to netilmicin. Twelve out of the total of 44 strains were multiply-res. (resistant to penicillin, streptomycin and tetracyclines), and 10 of these were also resistant to methicillin (M). All phage groups/complexes were represented, group III by 13, and the 83A complex by seven strains. Multiply-res. strains, resistant to both M and G were all resistant to mercury, but sensitive to arsenate, whereas such strains resistant to either M or G usually have been found resistant to both metals. Twenty-three strains lost resistance to G upon storage, among them only one multiply-res. Loss of resistance did not influence the metal resistance pattern. From one patient only, the various isolates (nine) differed in respect to bacteriological properties. It was concluded, however, that they all were descendants of the same unstable strain. In the majority of the cases treatment with G had preceded isolation of the resistant strain.

Two hundred and forty Danish patients with cystic fibrosis (97% of the total CF population in Denmark) participated in a point-prevalence study of Pseudomonas aeruginosa infection. One hundred and ninety-two patients were treated at the Danish CF centre and 48 patients were treated in other places. The age distribution was significantly different as no patients older than 19 years were found in the non-centre group. Pathogenic bacteria were isolated from the sputum of 96% of the patients. P. aeruginosa was more prevalent in patients from the centre, whereas Staphylococcus aureus was more prevalent in the non-centre group. No difference in serogroup and phage pattern of P. aeruginosa was found. There was a tendency that non-centre treated patients acquired chronic broncho-pulmonary P. aeruginosa infection later, but at the age of 16 years 90% of all patients will be chronically infected. Chronic P. aeruginosa infection was significantly more common in the age group 10-14 years at the centre than outside the centre. It is not possible to prevent chronic P. aeruginosa infection in CF patients treated in small groups and because of the better prognosis of centralized treatment the latter must be recommended.

Early in 1983 an epidemic of a Pseudomonas aeruginosa resistant to aminoglycosides, carbenicillin, ureidopenicillins, ceftazidime, cefsulodin and imipenem occurred in a cystic fibrosis centre. Most of the epidemic could be attributed to a specific nosocomial strain by means of O-grouping and phage-typing. This strain was present in the centre at a low frequency in 1973 and developed resistance during courses of chemotherapy. The epidemic was stopped by isolating patients with the resistant strains. Restrictive and selective use of antibiotics have not been sufficient to eradicate the resistant strains, which persist in 42% of the patients. The extensive use of the third generation cephalosporins in the clinic is probably responsible for inducing and selecting for the resistant strains. Clustering of patients in the centre has facilitated the spread. First-line use of older beta-lactam antibiotics, close bacteriological monitoring and prompt isolation of patients with resistant strains are recommended.

In the period 1969-1974 a decreasing number of Danish hospital departments recorded epidemic occurrences of Staphylococcus aureus strains, and the local spread of strains was less extensive. Multiple-resistant strains of the 83 A complex were succeeded by type 94 strains, resistant to penicillin only. The level of methicillin resistance fell from 19 to 6%. 12% of methicillin resistant strains are now sensitive to streptomycin and/or tetracyclines; they do not represent a few clones, but belong to a wide spectrum of phage types. The changes follow a reduction in the consumption of streptomycin and tetracyclines not of methicillin or other penicillins. As a contrast to the general reduction of combined resistance to streptomycin and tetracyclines, the strains in dermatological departments, where tetracyclines are commonly used, maintain a high degree of resistance to tetracyclines alone.

A comparative study was made on tobramycin combined with either carbenicillin (500 mg/kg/day) or one of the new penicillins: azlocillin or piperacillin (both 300 mg/kg/day) in 50 cystic fibrosis patients with chronic Pseudomonas aeruginosa infection. Average 2-h levels of penicillins in serum were 46 micrograms/ml (piperacillin), 88 micrograms/ml (azlocillin) and 66 micrograms/ml (carbenicillin). The Pseudomonas strains were significantly more sensitive to piperacillin and azlocillin than to carbenicillin (minimal inhibitory concentrations 1.9, 2.3 and 4.2 micrograms/ml). In 21 of 54 treatment course temporary eradication of Pseudomonas was achieved. Improved ventilatory capacity and diminished proteolytic activity in sputum were seen in most patients with or without bacteriological treatment success. Resistant strains - often belonging to other types - appeared in the patients with treatment failure. With increased number of precipitating antibodies against Pseudomonas and with increased minimal inhibitory concentrations, the chance of eradication was smaller. Seven out of 20 treated with azlocillin and 14 out of 30 treated with piperacillin developed fever and exanthema by the end of treatment. Our experience suggests caution in the use of the new penicillins.

Hypotheses for the origin of multiply-resistant coagulase-negative staphylococci from 146 patients undergoing cardiac surgery were tested. All received cephalothin per-operatively. Antibiotic susceptibility testing, phage-typing, bio-typing, and test for Tween-80-splitting enzyme were used to characterize 132 isolates from nose swabs. Seventy-five percent of the pre-operative susceptible isolates were of biotype 1, while biotypes 3 and 4 made up 59% of the post-operative, multiply-resistant isolates. Fifty-three percent of the isolates were typable by phage-typing. Typability of isolates of biotype 1 was high (56%) while almost 75% of biotype 4 were untypable. Susceptible isolates were more often typable than multiply-resistant ones. Of the 146 patients, 105 (72%) carried coagulase-negative staphylococci pre-operatively, only two of them carried multiply-resistant strains. Fifty-nine patients (41%) were colonized with multiply-resistant coagulase-negative staphylococci post-operatively. By combining the results of bio-typing, phage-typing, and test for Tween-splitting enzyme the study made it probable that a maximum of ten patients (6.8%) already carried multiply-resistant strains on admission to the hospital or were carriers of initially susceptible strains which developed multiple-resistance during administration of antibiotics. It therefore seemed likely that most of the patients were deprived of their natural bacterial flora by antibiotic treatment and subsequently colonized post-operatively with multiply-resistant coagulase-negative staphylococci from the environment.

Clinical and bacteriological information on Staphylococcus aureus endocarditis from hospitals all over Denmark in the period 1976-1981 was reviewed in 119 cases, 61 females and 58 males. Patient ages ranged from 16 days to 85 years, with a median age of 63 years. The overall mortality was 71%. The mortality correlated significantly with such factors as age, hospital-acquired infections and resistance to penicillin in infecting strains. Hospital-acquired infections occurred in 38% of the patients. The distribution of phage types among strains isolated from blood cultures from patients with endocarditis corresponded to that of strains from other septicaemia cases. Group I and group III strains and strains of the 94, 96 complex comprised 74% of the phage types of the present material. Infections of the skin were the most common portal of entry for the infecting strains. Apart from drug addicts, of which 11 cases were included, mortality did not correlate with the presence of any underlying diseases.