Pervasive fungicide resistance in Botrytis from strawberry in Norway: Identification of the grey mould pathogen and mutations

Control of grey mould, caused by Botrytis spp., is a major challenge in open field strawberry production. Botrytis was isolated from plant parts collected from 19 perennial strawberry fields with suspected fungicide resistance in the Agder region of Norway in 2016. Resistance to boscalid, pyraclostrobin and fenhexamid was high and found in 89.1%, 86.0% and 65.4% of conidia samples, respectively. Multiple fungicide resistance was common; 69.6% of conidia samples exhibited resistance to three or more fungicides. Botrytis group S and B. cinerea sensu stricto isolates were obtained from 19 and 16 fields, respectively. The sdhB, cytb, erg27 and mrr1 genes of a selection of isolates were examined for the presence of mutations known to confer fungicide resistance to boscalid, pyraclostrobin, fenhexamid and pyrimethanil plus fludioxonil, respectively. Allele-specific PCR assays were developed for efficient detection of resistance-conferring mutations in cytb. Among B. cinerea isolates, 84.7%, 86.3% and 61.3% had resistance-conferring mutations in sdhB, cytb and erg27, respectively. A triplet deletion in mrr1, resulting in ΔL497, commonly associated with the multidrug resistance phenotype MDR1h, was detected in 29.2% of Botrytis group S isolates. High frequencies of resistance to several fungicides were also detected in Botrytis from both imported and domestically produced strawberry transplants. Fungicide resistance frequencies were not different among fields grouped by level of grey mould problem assessed by growers, indicating factors other than fungicide resistance contributed to control failure, a fact that has important implications for future management of grey mould.     

Modeling and Simulation of Food Products in Superchilling Technology

The food superchilling process is of increasing importance because of its benefit in achieving food quality and extending shelf life of food products. The rate of the superchilling process is critical to the products’ quality and to the productivity of the process, and therefore the superchilling dynamics are of extreme importance. The objective of this work was to develop a one-dimensional implicit finite difference numerical model for predicting partial freezing time necessary to achieve an optimal degree of superchilling in foods and to validate the model experimentally. The evaluation of degree of superchilling was determined using finite slab and measured by using a calorimetry method. There is a good level of agreement between numerical simulation and laboratory experimental results.     

A mycobacterial coinfection in a dog suspected on blood smear

A 4‐year‐old neutered female crossbred Shepherd was referred for a history of 10 days of anorexia, polyuria, polydipsia, polyadenomegaly, and diarrhea. On physical examination, the dog appeared quiet, responsive, and apyretic, with generalized and severe lymphadenomegaly. Hematologic abnormalities included neutrophilic leukocytosis with left shift, and lymphopenia. Blood smears revealed intracytoplasmic bacilli negatively stained with May–Grünwald–Giemsa in neutrophils and monocytes. Lymph node smears revealed pyogranulomatous adenitis with calcified deposits and many negative‐staining rod structures, both within the cytoplasm of neutrophils and macrophages, and free in the background. An acid‐fast stain (Ziehl–Neelsen) confirmed the diagnosis of mycobacterial infection. The dog was euthanized for public health and ethical reasons, and the postmortem examination revealed severe and generalized granulomatous and necrotizing lymphadenitis, panniculitis, and hepatitis, and infiltration of epithelioid macrophages in the lungs, colon, and spleen. Numerous acid‐fast bacilli, consistent with mycobacterial infection, were observed both in the cytoplasm of epithelioid macrophages and giant cells, and free in the background. Mycobacterium bovis was first confirmed by conventional PCR of organ extracts. Mycobacterium avium was detected in a culture of the same organs. Further PCR amplifications and sequencing revealed a coinfection with 2 different species of mycobacterium, one belonging to the Mycobacterium avium complex and the other to the Mycobacterium tuberculosis complex.     

Occurrence of weak mutators among avian pathogenic Escherichia coli (APEC) isolates causing salpingitis and peritonitis in broiler breeders

A collection of 46 avian pathogenic Escherichia coli (APEC) isolates was examined for the presence of mutators by determining the rate of mutation to rifampicin resistance. The collection included 34 E. coli isolates obtained in pure culture from chronic lesions of salpingitis and peritonitis in 34 broiler breeders, of which 12 were associated with the development of secondary septicemia. Twelve additional isolates were obtained from a clonal outbreak (ST95) of E. coli peritonitis syndrome (EPS), the lesions of which changed gradually over time into a subacute/chronic form. The hypothesis of the present study was that mutation rates would be higher for chronic infection isolates than for isolates from acute infections/exacerbations. The distribution of mutation rates followed a pattern similar to that found for other clinical isolates of E. coli, with a modal/median value of 1.47 × 10⁻⁸. Of the 46 isolates, 24% (n = 11) were weakly hypermutable (2.00 × 10⁻⁸ ≤ μ < 2.00 × 10⁻⁷), however, no strong mutators were detected (μ ≥ 2.00 × 10⁻⁷). Chronic salpingitis isolates had the highest proportion (45%, P = 0.001) of weak mutators and also, significantly higher mutation rates (P = 0.003) compared to isolates that caused septicemia (4%). In addition, mutation rates were significantly lower among ST95 isolates (P < 0.0005), and among isolates from the same clonal group as ST95 (P = 0.027), when compared to isolates from other groups. Although a clear association with the time phase of infection (as lesions of EPS became more chronic) could not be observed (ρ = 0.523, P = 0.081), a higher frequency of weak mutators among chronic infection isolates suggests that increased mutation rates play a role in adaptation of APEC to long-term persistence in an infected host environment.     

Ecology, history and silviculture of Scots pine (Pinus sylvestris L.) in western Norway - a literature review

Results from a literature review on pinewood ecology, silviculture,genetics, aspects of history and forest resources of Scots pine(Pinus sylvestris L.) in western Norway are presented. The pinewoodscover 40 per cent of the forested land, 0.31 million ha. Duringthe last 75 years, the area has increased by 17 per cent andthe growing stock has risen from 10 to 34 million m³. The impactof man in previous times was very marked, and has had a significantinfluence on the present forest conditions. The pronounced climaticgradients mixed with the topographic variation – fromthe coastal plains via the fjord systems to the high mountains– is reflected in rather steep gradients in the pine forestvegetation. Various floristic elements can be distinguished,from oceanic via the suboceanic in the outer islands to thethermophytic, boreonemoral and boreal elements in the innerfjord districts and valleys. The introduction of spruce (Piceaspp.) plantations on 10–15 per cent of former native pineforests has not negatively affected the bird fauna at the landscapescale. Although not particular species rich, the pine forestsharbour species usually not found in other forest types. Sofar, most work in the field of silviculture and forest ecologyin the pinewoods of West Norway has been in the form of casestudies. Implications of the results for forestry in the regionare briefly discussed.     

Tolerance to Acid Water Among Strains and Life Stages of Atlantic Salmon (Salmo Salar L.)

Reintroduction of Atlantic salmon (Salmo salar L.) after liming of acidified barren salmon rivers could benefit by choosing acid tolerant strains. Testing different life history stages from fry to smolts of five salmon strains with different acidification history demonstrated strain-specific variation in tolerance to acid aluminum-rich waters for stages from fry to parr. Contrary to expectation, salmon from non-acidified rivers were more tolerant. Differences in sensitivity were found between life history stages. Within a specific life history stage, size-dependent sensitivity was found; small fish being more sensitive to pH while large fish were more sensitive to aluminum. Presmolts showed the same relative tolerance between strains as younger stages. These differences disappeared, however, when the smolt reached full smoltification, probably due to supersensitivity at this stage. Poor water quality during the last period of smoltification and outmigration can thus mask the genetic potential for tolerance to acidic rivers.     

Genetic diversity and associated pathology of Pasteurella multocida isolated from porcine pneumonia

Pasteurella multocida is a widespread respiratory pathogen in pigs associated with atrophic rhinitis and contributing to aggravation of the pulmonary lesions. The aims of the present study were to characterize isolates of P. multocida from porcine bronchopneumonia by pulsed-field gel electrophoresis (PFGE), PCR based capsular typing and multilocus sequence typing (MLST) and to compare clonal complexes outlined with the type of histological lung lesions to investigate if a correlation between clonal lineages and lesions might exist. Isolates of P. multocida were obtained from cases of cranioventrally located porcine bronchopneumonia. All lung lesions were described and classified according to histological lesions. A total of 139 isolates, from lung (n=111), pericardial sac (n=21) and kidney (n=7) of 111 pigs were described using PFGE with ApaI as the restriction enzyme. Furthermore, 20 and 29 isolates were characterized by capsular serotyping and multilocus sequence typing, respectively. PFGE demonstrated 15 different clusters showing 50% or more similarity. All selected isolates were of capsular serotype A and only three main sequence types (ST) were detected among the isolates. Associations were not found between histopathology and clonal complexes of P. multocida. In conclusion, PFGE demonstrated a high diversity of genotypes of P. multocida associated with porcine bronchopneumonia. However, isolates obtained mainly belonged to few STs, indicating that isolates of P. multocida associated with porcine bronchopneumonia originates from a limited number of clonal lineages and therefore might have adapted to porcine hosts. No correlation was demonstrated between genotypes and types of lesions, and extra-pulmonary spreading was only rarely demonstrated.     

Clonality of Enterococcus faecalis associated with amyloid arthropathy in chickens evaluated by multilocus sequence typing (MLST)

The aim of the study was to evaluate the clonality of 21 strains of Enterococcus faecalis associated with arthritis and amyloid arthropathy by multilocus sequence typing (MLST). The strains originated from five countries. Fifteen of the 21 strains demonstrated the same sequence type, ST82, including a reference amyloid arthropathy strain from the Netherlands. Some of the demonstrated sequence types (ST36, ST59 and ST82) have also been described from human clinical samples while other types are reported for the first time. The results confirm previous observations that outbreaks of amyloid arthropathy seem to be clonally related and indicate a wide distribution of the predominant sequence type which was demonstrated in four countries in Europe and in the USA.     

Effect of oxygenation, aeration and pH control on water quality and survival of turbot, Scophthalmus maximus (L.), kept at high densities during transport

The survival of turbot, Scophthalmus maximus (L.), kept at high densities (0.5 kg 1^?1 water) was studied when O₂ was added by oxygenation, aeration, a combination of oxygenation and aeration, or by aeration with control of water pH by the addition of acid. Oxygen instead of air increased the LT₅₀ (lethal time for 50% of the population) from 40 h to 130 h. Aeration in combination with acid addition further increased LT₅₀ to 220 h. The results show that the transport time of live fish can be increased by reducing pH, either by raising carbon dioxide levels or by adding acid, probably because of reduced toxicity of total ammonia (NH₃ and NH₄⁺). In all groups, mortality was recorded at NH₃ concentrations above 0.4 mg 1^?1 NH₃-N.     

Differences between Atlantic halibut (Hippoglossus hippoglossus L.) and turbot (Scophthalmus maximus L.) in tolerance to acute low temperature exposure

Juvenile Atlantic halibut (Hippoglossus hippoglossus L.) and turbot (Scophthalmus maximus L.) acclimated to 8–9 °C sea water were transferred to sea water at 1 °C for 7 days. An immediate spasm-like response was noticed in the turbot after the transfer, but not in the halibut. None of the fish died. The turbot suffered from substantial physiological disturbances after the transfer. Their blood plasma Cl^– concentration increased and muscle water content decreased markedly. The plasma glucose concentration was several times higher than the control level from day one onwards. A decrease in the haematocrit was recorded some hours after transfer, followed by a marked increase on day 7. In halibut, the plasma Cl^– concentration fell somewhat during the first days, but returned thereafter to its pretransfer level. The muscle water content was unchanged. Both haematocrit and plasma glucose concentration were unchanged until day 7, when both were significantly higher. The differences in the response to low-temperature challenge between halibut and turbot are probably related to genetic differences in temperature tolerance, which reflect differences in the distribution of the species.