Aerobic microbial respiration in 86-million-year-old deep-sea red clay

Microbial communities can subsist at depth in marine sediments without fresh supply of organic matter for millions of years. At threshold sedimentation rates of 1 millimeter per 1000 years, the low rates of microbial community metabolism in the North Pacific Gyre allow sediments to remain oxygenated tens of meters below the sea floor. We found that the oxygen respiration rates dropped from 10 micromoles of O(2) liter(-1) year(-1) near the sediment-water interface to 0.001 micromoles of O(2) liter(-1) year(-1) at 30-meter depth within 86 million-year-old sediment. The cell-specific respiration rate decreased with depth but stabilized at around 10(-3) femtomoles of O(2) cell(-1) day(-1) 10 meters below the seafloor. This result indicated that the community size is controlled by the rate of carbon oxidation and thereby by the low available energy flux.     

Diversity of Flavobacterium psychrophilum and the potential use of its phages for protection against bacterial cold water disease in salmonids

Flavobacterium psychrophilum causes rainbow trout fry syndrome (RTFS) and cold water disease (CWD) in salmonid aquaculture. We report characterization of F. psychrophilum strains and their bacteriophages isolated in Chilean salmonid aquaculture. Results suggest that under laboratory conditions phages can decrease mortality of salmonids from infection by their F. psychrophilum host strain. Twelve F. psychrophilum isolates were characterized, with DNA restriction patterns showing low diversity between strains despite their being obtained from different salmonid production sites and from different tissues. We isolated 15 bacteriophages able to infect some of the F. psychrophilum isolates and characterized six of them in detail. DNA genome sizes were close to 50 Kbp and corresponded to the Siphoviridae and Podoviridae families. One isolate, 6H, probably contains lipids as an essential virion component, based on its chloroform sensitivity and low buoyant density in CsCl. Each phage isolate rarely infected F. psychrophilum strains other than the strain used for its enrichment and isolation. Some bacteriophages could decrease mortality from intraperitoneal injection of its host strain when added together with the bacteria in a ratio of 10 plaque-forming units per colony-forming unit. While we recognize the artificial laboratory conditions used for these protection assays, this work is the first to demonstrate that phages might be able protect salmonids from RTFS or CWD.     

Evaluation of a simple means to reduce discard in the Kattegat-Skagerrak Nephrops (Nephrops norvegicus) fishery: Commercial testing of different codends and square-mesh panels

Discarding of fish species in the Kattegat-Skagerrak Nephrops directed fisheries remains at high levels. In this work we have tested four sets of codends pair-wise with the catch comparison technique under full commercial conditions to assess their potential in reducing the catch of undersized commercial species, in particular cod. We investigate the position of a 90 mm square-mesh panel (SMP), the effect of increasing the mesh size from 90 to 120 mm in the SMP, the effect of inserting a 90 mm SMP in an 80 mm codend and the effect of increasing the codend mesh size from 90 to 120 mm in a total of 89 hauls. Inserting the SMP 3–6 m from the codline compared to 6–9 m from the codline, reduced catches of cod above the minimum landing size (MLS) and Nephrops above and below the MLS. Increasing the mesh size in the SMP from 90 to 120 mm increased the catch of cod above the MLS in weight with an overall increase of 12% in the revenue. The effect of installing a 90 mm SMP 6–9 m from the codline in a nominal 80 mm codend had no effect on the catch of cod. Increasing the nominal codend mesh size from 90 to 120 mm reduced the catch of all species below the MLS, except monkfish. Catches of Nephrops above the MLS were, however, reduced by approximately one third and the total loss in revenue was 21%.     

Dietary calcium-phosphorus ratios for growing pigs in relation to serum levels and bone development

Two experiments comprising a total of 80 Danish Landrace pigs in the period 20–90 kg live-weight were carried out to study the effect of dietary levels and ratios of Ca and P on serum values of Ca, inorg. P, and alkaline phosphatase and on the development of degenerative arthritis and atrophic rhinitis.In both experiments each kg feed (Table 1) was supplemented with 600 i. u. vitamin D₃ and 80 p. p. m. Zn.In the first experiment the Ca:P ratio was 1.2 and the amount of Ca increased from 0.48 % to 1.20 % without influencing daily gain, feed conversion, carcass performance (Table 2) or frequency of atrophic rhinitis (Table 3). The Ca:P ratio in scapula was 2.15. Only when the diet was supplemented with 1.20 % Ga and 1.00 % P the bone quality was histomorphologically adequate (Figs. 1–4).In the second experiment various Ca:P ratios (ranging from 0.1 to 3.0) were compared with the recommended level and ratio of 0.72 % Ga and 0.60 % P. Performance of pigs on the normal level was better than for the other pigs (Table 4). Clinically, various degrees of leg-weakness and -deformation occurred, most pronounced in the high Ca — low P group. Very little lameness and no convulsions were observed. Marked hypocalcemia was not found. Unilateral high Ca intake increased serum Ca significantly, and serum alkaline phosphatase slightly, and decreased serum P (Figs. 5–7). Advanced pericytic osteolysis, poor mineralization and generalized osteodystrophia fibrosa were found in the low Ca groups (Figs. 8–13). The groups showed no differences in frequency of atrophic rhinitis (Table 5), and rickets was not observed.The incidence of degenerative joint lesions was not increased in pigs on the unfavourable Ca:P ratios, and depended apparently more on litters than on diets.     

Detection of papillomavirus-DNA in mesenchymal tumour cells and not in the hyperplastic epithelium of feline sarcoids

Abstract We examined 12 formalin-fixed paraffin-embedded feline skin tumours which had the histopathological features of fibropapillomas for the presence of papillomavirus (PV) DNA using touchdown polymerase chain recation (PCR), DNA sequencing and nonradioactive in situ hybridization. Nine of the tumours contained a 102-bp PCR product demonstrated using consensus PV primers that amplify a portion of the L1 gene. The nucleotide sequences are closely related, but not identical to that of ovine PV type 2, rabbit oral PV and reindeer PV. The deduced amino acid sequences had strong homologies with the major capsid protein L1 of deer PV, bovine papillomavirus (BPV) 1 and BPV 2, and European elk PV. Although PV antigens were not detected in any of the tumours by immunohistochemistry, PV DNA was demonstrated in individual mesenchymal cells or cell nests of 4/12 tumours by in situ hybridization. A nonproductive infection of mesenchymal fibroblast-like tumour cells with a papillomavirus would explain the lack of PV antigen expression and the absence of PV DNA in the hyperplastic epithelium. Because these tumours and their pathogenesis are similar to equine sarcoids, we suggest that they should be reclassified as 'feline sarcoids' instead of fibropapillomas.     

Evaluation of a single dose versus a divided dose regimen of danofloxacin in treatment of Actinobacillus pleuropneumoniae infection in pigs

A single versus a divided dose regimen of danofloxacin was evaluated in treatment of porcine Actinobacillus pleuropneumoniae infection using clinical observations combined with biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Twenty hours after experimental infection, the 18 pigs received danofloxacin intravenously as a single dose of 2.5mg/kg or four doses of 0.6 mg/kg administered at 24h intervals. These dosage regimens resulted in similar AUCs of the plasma danofloxacin vs time curve. The maximum concentration was 3.5-fold higher using the single dose regimen, while the time with concentrations above the MIC was 2.5-fold longer using the fractionated regimen. Using the single dose regimen, temperature was normalised 32 h post-infection. In contrast, normalisation was delayed until 44 h post-infection using four low doses and a relapse with elevated temperatures at 52 and 68 h was observed. No other significant differences between the treatments were found, neither regarding clinical, haematological nor biochemical observations. The use of the more convenient single dose regimen was appropriate, as it was at least equivalent to the fractionated regimen.     

Comparisons of sampling procedures and time of sampling for the detection of Salmonella in Danish infected chicken flocks raised in floor systems

Bacteriological follow-up samples were taken from 41 chicken (Gallus gallus) flocks in floor systems, where Salmonella enterica (Salmonella) had been detected either directly in bacteriological samples or indirectly by serological samples. Three types of follow-up samples were compared to each other within each flock: 1) 5 pairs of socks, analysed as 5 samples, 2) 2 pairs of socks, analysed as one sample, and 3) 60 faecal samples, analysed as one pooled sample. Agreement between sampling methods was evaluated by the following statistical tests: 'Kappa', 'The adjusted rand', McNemar's test for marginal symmetry, Proportion of agreement P0, P+, P-, and Odds Ratio. The highest agreement was found between the 2 types of sock sampling, while the lowest agreement was found by comparing 60 faecal samples with 5 pairs of socks. Two pairs of socks analysed as one pool appeared to be just as effective in detecting S. enterica as the 60 faecal samples. In broiler flocks, 5 pairs of socks were used both in the routine samples taken at about 3 weeks of age for the establishment of infection of the flock, and as one of the follow-up samples taken shortly before slaughter age, which means that the only notable differences between the 2 sampling rounds were the age of the broilers and of their litter. S. enterica was detected more frequently in samples from broilers about 3 weeks old, than in similar samples taken from broilers a few days prior to slaughter at ca. 33-40 days of age.     

Maximum-likelihood estimation of sensitivity and specificity of ELISAs and faecal culture for diagnosis of paratuberculosis

The accuracy of three diagnostic tests for paratuberculosis was evaluated using maximum-likelihood estimation of sensitivity and specificity. We also explored the variety of estimates that can be obtained if the tests are to be used in populations of different composition with regard to infection and disease states. Two enzyme-linked immunosorbent assays (ELISAs) were evaluated separately with the faecal culture (FC). The study was carried out as a cross-sectional field study to cover all likely states of infection with Mycobacterium avium subsp. paratuberculosis.The three basic assumptions for the maximum-likelihood technique were evaluated to validate the results. Our accuracy estimates for the ELISAs were not very different from those previously published, but those for faecal culture differed if a different cut-off value was chosen for the ELISA. If faecal culture was used for screening in a Danish dairy region where the median ELISA reading was a measure of the general disease situation, the sensitivity of the faecal culture was 20-25%. If faecal culture was used as a confirmatory test on cows with a high ELISA reading (and thus high level of antibodies), the sensitivity of the faecal culture would be in the range 60-70%. These results emphasise the importance of the composition of a target population before selecting a specific diagnostic test for a given purpose. We concluded that faecal culture is useful for confirmation but not for screening purposes.     

Antimicrobial susceptibility and presence of resistance genes in staphylococci from poultry

The species distribution, susceptibility to 19 antimicrobial agents and presence of selected genes encoding resistance to macrolides, streptogramins and tetracyclines were examined among 118 staphylococcal isolates from infections of poultry in Denmark. Isolates were identified using a combination of conventional biochemical testing and 16S rDNA sequencing. The most common species were Staphylococcus aureus (83), Staphylococcus hyicus (11), Staphylococcus xylosus (9) and Staphylococcus cohnii (6). The isolates were susceptible to most antimicrobials tested. A high frequency of S. aureus (30%) was resistant to ciprofloxacin. Only six (7%) S. aureus isolates and one Staphylococcus saprophyticus were penicillin resistant. Resistance to sulphamethoxazole was observed among 16 (19%) of S. aureus isolates and two coagulase negative staphylococci (CNS). Twenty (24%) of the S. aureus isolates were resistant to erythromycin and 19 of these isolates contained the ermA gene, whereas the remaining isolate contained the ermC gene. Eleven (48%) of the novobiocin resistant CNS were resistant to erythromycin and all these isolates contained the ermA gene. Two isolates identified as S. xylosus, were found to be resistant to streptogramins and both contained the vatB- and the vgaB-genes. Thirty-nine (47%) of the S. aureus isolates, three of nine S. hyicus and eight of the 23 novobiocin resistant CNS were tetracycline resistant and all contained the tet(K) gene. A single S. aureus isolate also contained the tet(M) gene. The present study showed a frequent occurrence of resistance to fluoroquinolones, tetracycline and macrolides among staphylococci isolated from broilers in Denmark, whereas the occurrence of resistance to other antimicrobial agents remains low. Similar genes, encoding resistance to erythromycin, tetracycline and streptogramins to those previously observed, were detected.