Non-abscised aborted sweet cherry fruits are vulnerable to fruit decaying fungi and may be sources of infection for healthy fruits

During three growing seasons (1999–2001), disease incidence of non-abscised aborted and normally developing (sound) sweet cherry fruits were investigated in a research orchard at Ullensvang, western Norway. To reveal possible fungal infections, aborted and sound fruits of two cultivars (cv. Van for three years and cv. Lapins for two years) were harvested over four to seven weeks in an experimental orchard during the green fruit phase and incubated at 20°C in moisture saturated air for 7 days. The most frequently observed fungal pathogens were Monilinia laxa, Colletotrichum gloeosporioides, and Botrytis cinerea. The mean of all observations (±standard deviation) over three years, showed that aborted and sound fruits had a disease incidence of 51.9% (±31.8) and 5.2% (±9.7), respectively, after seven days incubation. In 24 of 25 trials aborted fruits had significantly higher disease incidence than sound fruits after incubation. In one season, when fruits were collected from two commercial orchards, aborted fruits had much higher disease incidence than sound fruits (a mean of 6.5 and 4.5 times higher incidence for the two orchards, respectively). Time of fruit abortion varied with the years (two years observation) and the two cultivars, but the major abortion took place between the fourth and eighth week after anthesis. A higher disease incidence and more rapid disease development in non-abscised aborted fruits indicate that they are more vulnerable to fungal colonisation than sound fruits and may thus be potential incoulum sources for neighbouring, healthy fruits.     

Evaluation of β-(1 → 3, 1 → 6)-glucans and High-M alginate used as immunostimulatory dietary supplement during first feeding and weaning of Atlantic cod (Gadus morhua L.)

Stimulation of the non-specific defence enhances the disease resistance and growth, and has good potentials as a measure for increased microbial control in juvenile production of marine fish and shellfish. So far, the most commonly used immunostimulants are β-(1 → 3, 1 → 6)-glucans, and in this study the stimulatory potential of a β-(1 → 3, 1 → 6)-glucan of marine origin, the storage polysaccharide from the marine diatom Chaetoceros mülleri, was examined. The glucan (chrysolaminaran) was extracted from cultures of C. mülleri, and used as a dietary supplement in two first feeding experiments with larvae of Atlantic cod Gadus morhua L. In one experiment the microalgal glucan was compared to the commercial yeast-glucan product MacroGard®, and in the other to an alginate with a high content of mannuronic acid (High-M alginate) isolated from Durvillaea antarctica. The stimulants were given via rotifers, and weaning to formulated feed was initiated at day 17 or 18 after hatching. The survival ± SEM at day 27 after hatching was 24.5 ± 2.0%, 14.8 ± 4.5% and 13.1 ± 1.4% for the groups fed C. mülleri-glucan, yeast glucan and for the control, respectively, in the first experiment. The group fed C. mülleri-glucan group had higher survival compared to the control (P < 0.05) group, whereas the yeast glucan had no positive effect on the survival (p > 0.05). The dry weights of the groups at day 27 were low, with 203.2 ± 52.2, 165.2 ± 43.4 and 198.5 ± 58.1 μg per larva for the C. mülleri-glucan, yeast glucan and control groups, respectively. In the second experiment the survival in the period of feeding formulated feed (days 18-30) were 44.6 ± 4.3%, 44.7 ± 1.3%, and 33.8 ± 4.1% survival for the C. mülleri-glucan, High-M alginate and control group, respectively. The cod larvae fed C. mülleri-glucan reached an average weight of 531.6 ± 17.2 μg at day 30, which was significantly higher (p < 0.05) than the control group that had an average of 473.6 ± 3.5 μg. The larvae fed High-M alginate had an average weight of 470.3 ± 31.6 μg per larva at day 30, and not significantly different from the control (p > 0.05). The early weaning to formulated diet had detrimental effect on the growth of the larvae. In both experiments the C. mülleri-glucan group was the only group showing a positive growth rate in the period of weaning to dry feed. The microbial conditions in larval gut and water were monitored with respect to total colony forming units on Marine agar, and Vibrio- and Pseudomonas-like species on selective agars (TCBS and marine Pseudomonas Agar with CFC-supplement). The larvae were rapidly colonised after hatching, but no or weak effects of the stimulants were observed on the colonisation rates or the composition. The total CFU varied from 10¹ to 10² CFU per μg larva after initiation of the first feeding. The percentages of Pseudomonas-like bacteria increased throughout the period, whereas the levels of Vibrio-like bacteria were low and stable. The chrysolaminaran from the diatom C. mülleri was shown to be a promising candidate for use as an immunostimulatory feed additive, and which should be further explored.     

Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae

Theaim of this study was to determine whether it is possible to influence thespecies composition of the gut microflora in 70-days old halibut larvae(Hippoglossus hippoglossus L.) by addition of bacteriabioencapsulated in Artemia franciscana in two shortpulses.Two Vibrio strains, PB 1-11 and PB 6-1 were used. Twotreatments received each strain alone, a third treatment received the twobacterial strains in mixture, whereas in a control treatment no bacteria wereadded. Five fish tanks were included in each treatment. The bacteria werebioencapsulated in Artemia franciscana metanauplii duringashort-term incubation in bacterial suspensions of the specific strains, andwereadded in two pulses, on day 1 and day 10 of the experiment. Addition ofbacteriadid not increase the total number of colony-forming units (CFU) in the larvalgut. The total CFU in the water was lower in tanks added bioencapsulatedbacteria than in the control treatment (p < 0.05). Animmunocolony blot assay was used to measure the numbers of the specificbacteriaadded in samples of water, A. franciscana, and fishlarvae.At the end of the experiment on day 13, the strain PB 1-11 reached on average10% of the total CFU when added alone and 25% when added in mixture with PB6-1.Strain PB 6-1 reached at the end of the experiment 26.8% of the total CFU whenadded alone and 24.2% when added in mixture with PB 1-11, whereas thepercentages of PB 1-11 and PB 6-1 in the control fish were 12.7% and 10.8% ofthe total CFU, respectively. However, the differences between the experimentaltreatments compared with the control group were not significant(p > 0.05). The addition of bioencapsulated bacteria intwo pulses within a ten days period was therefore not sufficient to influencethe species composition of the microflora of the halibut larvae.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on sweet cherry leaves

Leaves of sweet cherry, exposed to either paraquat or freezing to quickly senesce the leaf tissue, were incubated in about 100% RH at 25°C for 6 d. Sporulating colonies of Colletotrichum acutatum, the cause of anthracnose, developed on up to 100% of the paraquat-treated and frozen leaves, and on none of the untreated controls. Number of leaves and leaf area containing C. acutatum on naturally infected leaves increased over time from May to September. Mean incidence of C. acutatum on leaf blades on fruit spurs and vegetative shoots from eight orchard/year samplings were 41 and 33%, respectively. Secondary conidiation (formation of short hyphae and new conidia) from conidia applied to detached leaves took place 6 h after inoculation, but only up to 3% of the conidia formed new conidia. It may be concluded that asymptomatic sweet cherry leaves frequently host C. acutatum and may be a potential inoculum source for cherry fruit.     

Inter‐individual variation in the dominant intestinal microbiota of reared Atlantic cod (Gadus morhua L.) larvae

The diversity of the microbiota in 15 Atlantic cod (Gadus morhua L.) larvae from three different hatcheries with variable food type and water treatment was characterized with culture dependent and independent approaches. The culture‐dependent approach was phenotypic characterization of bacterial isolates based on 16 tests, and the culture‐independent approach was terminal restriction fragment length polymorphism (T‐RFLP) of PCR amplified 16S rRNA. Both approaches revealed large inter‐individual differences in the microbiota of cod larvae. Data from real time PCR indicate substantially lower variation in bacterial number larva^?1 both within and between hatcheries than the data on colony forming units, resulting in an estimated four orders of magnitude variation in culturability between larvae. Considerably higher richness and diversity were observed with T‐RFLP than with phenotypic characterization, and no correlation between the two approaches was found for diversity indexes calculated at the individual level. Dissimilarity in the composition of the microbiota between individuals from the same rearing environment was comparable to the dissimilarities between hatcheries. The large inter‐individual variation is important to consider when designing experiments, as pooling of individuals will completely mask this variability. We conclude that colonization of individual cod larvae by bacteria seems to be a stochastic and not a deterministic process, which is not well understood.     

A NEW TYPE OF RADIOGRAPHIC BONE REMODELING IN THE DISTAL RADIAL AND ULNAR METAPHYSIS IN 54 NEWFOUNDLAND DOGS

In this longitudinal observational study, 118 growing Newfoundland dogs were followed with sequential radiographic examination of the right front limb, which were scheduled at ages 3, 4, 6, 12, 18, and 24 months to record evidence of skeletal changes. All affected dogs had evidence of changes at 6 months and dogs included in this study had radiographic examination at 6 months and at least two other scheduled examinations that included either 4 or 12 months. The dogs were privately owned and had individualized nutrition and environment with no maintenance protocol required by the project leaders. Irregularities in bone remodeling in the distal radius and ulna were seen in radiographs of 54 of 118 (45.8%) dogs. These irregularities differ from changes previously described in the Newfoundland dog. The reported irregularities were seen as islands of reduced opacity outlined by thickened, radiopaque osseous trabeculae, which were aligned with the axis of stress. All the 54 dogs with irregularities during some phase of development consistently had changes at 6 months of age. Reorganization of the changes in the bone was slow, and residual changes were visible at 18-24 months of age in many cases. The significance of these findings may lie in their potential for misinterpretation if they had been discovered in the presence of clinical signs such as any lameness or growth arrest. Radiographic appearance indicates disturbances in the bone maturation. The etiopathogenesis is unclear. Nutritional, environmental and genetic factors have to be studied.     

64,XX, SRY-, and ZFY-Negative Icelandic Horse Likely to Be True Hermaphrodite

A 4-year-old Icelandic horse, considered to be a mare, showed stallion-like behavior in a group of mares. On clinical examination, the horse turned out to have an enlarged erectable phallic clitoris. Ultrasound examination showed a normal-sized left ovary covered with numerous small follicular cysts and a compact testis-like tissue in place of the right ovary. The karyotype was normal for a mare (64,XX), and the horse was found to be negative for the Y chromosome–specific markers SRY, ZFY, and EIF1AY. This case indicates that the intersexual phenotype may be caused by autosomal recessive mutation, resulting in defects in cortisol biosynthesis rather than transferal of Y chromosome male–specific genes. This is the first report of an intersexual phenotype in an Icelandic horse that is likely to be a true hermaphrodite because of female sex chromosomes and a mixture of female and male gonads and external genitals.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on apple leaves

Fungi within the Colletotrichum acutatum species complex occur asymptomatically on plant parts of many different plant species. Leaves from apple orchards in southern Norway were sampled, frozen for five hours and incubated for six days to reveal presence of asymptomatic infections of C. acutatum. Number of leaves (incidence) and leaf area covered (severity) with conidial masses of C. acutatum were assessed biweekly on cv. Aroma from late May to late September during three growing seasons. The first finding of conidial masses occurred in the second half of July, and there was a higher incidence occurring in August and September. Sampling of leaves from fruit spurs and vegetative shoots of cvs. Aroma and Elstar showed that conidial masses of C. acutatum developed on leaves on both shoot types, and there was no difference in incidence between these two types. The fungus was detected on leaves from six of eight commercial orchards of cv. Aroma over three years, with a mean incidence of 5.5 %. After storage, bitter rot was found on apple fruit from all eight orchards. There was no correlation between incidence of conidial masses of C. acutatum on leaves and on fruit. In all orchards and seasons investigated, incidence and severity on leaves varied from 0 to 67 % and 0 to 85 %, respectively. The discovery of apple leaves containing conidial masses of C. acutatum clearly indicate for leaves as a potential source of inoculum for fruit infections.     

Apple rootstocks may become infected by Neonectria ditissima during propagation

Introduction and purpose: The ability of apple rootstocks to become infected by Neonectria ditissima, the cause of European canker, was studied over two years.

Materials and methods: Rootstocks B9 and M9 with a size suitable for grafting (6-10 mm stem diameter, termed rootstocks), and smaller sized rootstocks (<5 mm stem diameter, termed transplants) of B9, M9, M26, MM106 and Antonovka were inoculated with N. ditissima at different times, either with contaminated map pins or with spore suspensions. In addition, the rootstocks were either defeathered (side shoots removed), topped (top shoot headed) or both, to create wounds that would normally occur during propagation, while wounds on transplants were made by removing leaves.

Results and discussion: One month after inoculation, slightly sunken canker lesions had developed around the inoculation points of the map pins or wounds. No lesions developed on the non-inoculated controls. Map pin inoculation resulted in 30% to 89% infection and spore suspension sprayed on wounds from 5% to 45% infection. When the cankered areas were split open, brown lesions with necrotic tissue due to infection by N. ditissima appeared. The transplants of M9, M26 and MM106 inoculated with contaminated map pins in 2014 developed necrosis on 40% to 67% of the plants, but there were no differences in the incidence or severity among the different types. On the transplants of B9, Antonovka and M9 inoculated in 2015, there was more necrosis on B9 (42%) than on Antonovka (11%) and more sporulating lesions on B9 (29%) than on M9 (9%) or on Antonovka (4%).

Conclusion: It can be concluded that rootstocks used for apple trees may become infected by N. ditissima, and wounds should thus be protected during propagation.     

Size Distributions and Concentrations of Spheroidal Carbonaceous Fly-Ash Particles (SCPs) in Lake Sediments as an Aid to Detecting Locally Deposited Atmospheric Pollution

Size distributions and concentrations of spheroidal carbonaceous fly-ash particles (SCPs) > 5 μm have been counted in surface sediments from 20 lakes along a 62 km west-east transect in the Bergen area, Norway. Increased concentrations of SCPs are found in lakes immediately downwind, closest to the city. These lakes also have a dominance of larger particles (> 10 μm) than further downwind where smaller particles (5–10 μm) dominate. The pattern of SCPs > 5 μm is compared with a previous study of anthropogenic Pb and SCPs > 3 μm in the same sediments. Along the transect there is a difference in pattern between the concentrations of SCPs > 5 μm and the patterns of anthropogenic Pb and SCPs > 3 μm. The result indicates that increased altitude and rainfall probably explain the increased concentrations of anthropogenic Pb and SCPs > 3μm in the eastern part of the transect. Metals are in the size range for which atmospheric residence times are greatest, whereas particles > 5 μm in diameter are of the size thought to be effectively removed from the atmosphere by sedimentation. This may explain, at least partially, why there is a difference between the pattern of SCPs > 5 μm concentrations and anthropogenic Pb and SCPs > 3 μm concentrations. This study shows that SCPs > 5 μm in lake sediments can potentially provide a better signal of locally-deposited atmospheric pollution than trace elements from the same source. A SCP distribution dominated by large particles (> 10 μm) is found up to 20 km downwind of the city. Within this area, temporal SCP size distribution patterns can help detect local-deposited atmospheric pollution.