Plasma protein fractions in free-living white-tailed eagle (Haliaeetus albicilla) nestlings from Norway.

BACKGROUND: Capillary electrophoresis of plasma proteins has shown great potential as a complementary diagnostic tool for avian species. However, reference intervals for plasma proteins are sparse or lacking for several free-living avian species. The current study reports electrophoretic patterns and concentrations of plasma proteins determined for 70 free-living white-tailed eagle (Haliaeetus albicilla) nestlings from two locations in Norway (Steigen and Smøla) in order to establish reference values for this subpopulation using capillary electrophoresis. The nestlings were between 44 and 87 days of age, and the plasma protein concentrations were investigated for age, sex, year (2015 and 2016) and location differences. To our knowledge, this is the first report of reference intervals of plasma proteins analysed by capillary electrophoresis in free-living white-tailed eagle nestlings. RESULTS: The plasma protein concentrations (% of total protein, mean ± SE) were determined for prealbumin (13.7%, 4.34 ± 0.15 g/L), albumin (46.7%, 14.81 ± 0.24 g/L), α1-globulin (2.4%, 0.74 ± 0.03 g/L), α2-globulin (11.7%, 3.72 ± 0.06 g/L), ß-globulin (15.9%, 5.06 ± 0.08 g/L) and γ-globulin (9.6%, 3.05 ± 0.09 g/L). Significant differences were found between the two locations for prealbumin, α2- and γ-globulins. No significant differences were found between the two sampling years or sexes, and no effect of age was found for any of the plasma proteins. However, prealbumin levels were several folds higher than previously reported from adults of closely related birds of prey species. There were no other studies on capillary electrophoresis of nestling plasma available for comparison. CONCLUSION: Significant differences were found between sampling locations for prealbumin, α2- and γ-globulins, which may indicate differences in inflammatory or infectious status between nestlings at the two locations. Sampling year, sex or age had no significant effect on the plasma protein concentrations. These results provide novel data on plasma protein concentrations by capillary electrophoresis and may be useful for evaluation of health status in free-living white-tailed eagle nestlings.

Case report: Haemophagocytic histiocytic sarcoma in an english setter.

A 4-year-old English setter presented with a 1-week history of anorexia, lethargy and occasional vomiting. Blood analysis revealed moderate regenerative anaemia, mild monocytosis, thrombocytopaenia, hypoproteinaemia, hypoglobulinaemia, hypocholesterolaemia and increased C-reactive protein. On ultrasonography, the spleen had multifocal hypoechoic lesions. Fine needle aspirates from the spleen and liver showed marked extramedullary haematopoiesis, an increased number of histiocytes, haemosiderin deposits and erythrophagocytosis. A tentative diagnosis of haemophagocytic histiocytic sarcoma (HHS) was made, and the owners elected euthanasia. On autopsy, the liver and spleen were enlarged. The spleen had an uneven surface and a yellow-tan spotted appearance. Histologically, the red pulp was highly cellular and dominated by erythroid cells, as well as a population of larger polygonal cells and aggregates of histiocytes. HHS was confirmed by CD11d immunolabelling. This represents the first documented case of HHS in an English setter.

Haematological and serum biochemical values in Norwegian sled dogs before and after competing in a 600 km race.

BACKGROUND: Long-distance racing is known to cause alterations in haematological and serum biochemical parameters in sled dogs. Given that finishing status reflects the physical condition in dogs completing a race, such variations will mainly be the result of physiological adaption achieved during endurance exercise. However, changes observed in withdrawn dogs may indicate pathological conditions. The aim of this study was to reveal changes in haematological and serum biochemical values in sled dogs participating in a long-distance race, with emphasis on the withdrawn dogs. Sixty-five sled dogs participated in a clinical prospective cohort study: 46 dogs competed in the 600 km race (25 finishing and 21 withdrawn dogs), and 19 dogs served as controls. Blood sampling was performed early in the training season and after the race. RESULTS: When compared to control dogs, both withdrawn and finishing dogs showed significant increases in neutrophil count, C-reactive protein, blood urea nitrogen and sodium/potassium ratio. Significant decreases were found in erythrocytes and eosinophil cell count, and in haematocrit, haemoglobin, total protein, albumin, globulin, creatinine, potassium and calcium levels. Finishing dogs presented significant increases in white blood cells, large unstained cells, monocyte count and cortisol level compared to control dogs. In contrast, withdrawn dogs had significant elevations in alanine aminotransferase and alkaline phosphatase activity, as well as parameters associated with muscle metabolism, such as aspartate aminotransferase, creatine kinase and phosphorus concentration. CONCLUSIONS: Competing sled dogs experienced minor changes in blood parameters in general, mainly revealing the same pattern among withdrawals and finishers. This might indicate that numerous changes simply reflect physiological adaption due to endurance exercise. However, the serum concentration of muscle enzymes was significantly increased only in the withdrawals, and were well above reference ranges. This reflects muscle degradation, which could be the main cause of performance failure in some of the withdrawals.

Hematologic values in calves during the first 6 months of life.

BACKGROUND: Age-related changes in hematologic values are known to occur in many species. Few published studies include repeated measurements of hematologic parameters in calves during the first months of life. OBJECTIVE: The aim of the present study was to monitor hematologic values by sequential measurements from birth to 6 months of age in 15 healthy calves of the Norwegian Red breed, and compare the results to reference intervals for adult, lactating dairy cows. METHODS: Fifteen clinically healthy calves were sampled every week during the first 5 weeks of life and every month thereafter until 6 months of age. Hematologic values were measured using the ADVIA 120 hematology system. Reference intervals were determined for 75 healthy adult cows of the same breed. RESULTS: Compared with adult reference intervals, the MCV was lower and the RBC count was higher in calves throughout the investigation period. Hemoglobin concentration stayed largely within the adult reference interval. Mean MCHC was lower than adult values for 5 weeks, then increased and reached adult values by weeks 10-12. The mean lymphocyte count for calves reached adult reference values at weeks 6-8, and the mean monocyte count increased steadily until weeks 14-16. For most leukocytes, interindividual variation was larger during the first 5-8 weeks of life. The mean platelet count for calves was higher than the adult reference interval until weeks 19-21 of age. CONCLUSIONS: Age-specific reference intervals for calves from birth to 6 month of age are needed for RBC count, MCV, MCHC, red cell distribution width, and platelet and lymphocyte counts.

NCR1 is an activating receptor expressed on a subset of canine NK cells.

Defining NK cells has been challenging in many veterinary species. Although several groups have described putative NK cell populations, there is still no consensus on a definition of NK cells in the dog. In the present study, canine NK cells are characterized as CD3(-)GranzymeB(+) cells, further divided into a NCR1(+) and a NCR1(-) subset. All dogs examined displayed both subsets in blood, although of quite variable magnitude. Following vaccination an increase was observed in the CD3(-) NCR1(-) cell population in blood, but not in the CD3(-) NCR1(+) population. Non-B non-T cell cultures stimulated with IL-2 and IL-15 were dominated by CD3(-)GranzymeB(+) cells after approximately 2 weeks and a large proportion of the CD3(-)GranzymeB(+) cells expressed NCR1. IL-12 stimulation lead to a further upregulation resulting in an almost uniform expression of NCR1. The cultured cells expressed MHC class II, showed a variable expression of CD8 and were negative for CD4 and CD21. The cultures were able to kill known NK cell targets, and NCR1 was shown to be a major activating receptor. A large proportion of the NCR1(+) cells, but none of the NCR1(-) cells, produced IFNγ in response to IL-12 stimulation. These results show that NCR1 defines two subsets of canine NK cells, likely to represent different activation stages, and that NCR1 acts as an activating receptor on canine NK cells.

NCR1+ cells in dogs show phenotypic characteristics of natural killer cells.

No specific markers for natural killer (NK) cells in dogs have currently been described. NCR1 (NKp46, CD355) has been considered a pan species NK cell marker and is expressed on most or all NK cells in all species investigated except for the pig which has both a NCR1(+) and a NCR1(-) population. In this study peripheral blood mononuclear cells (PBMC) from 14 healthy dogs, 37 dogs with a clinical diagnosis, including a dog diagnosed with LGL leukemia, and tissue samples from 8 dogs were evaluated for NCR1(+) expression by a cross reacting anti bovine NCR1 antibody. CD3(-)NCR1(+) cells were found in the blood of 93 % of healthy dogs and comprised up to 2.5 % of lymphocytes in PBMC. In a selection of healthy dogs, sampling and immunophenotyping were repeated throughout a period of 1 year revealing a substantial variation in the percentage of CD3(-)NCR1(+) over time. Dogs allocated to 8 disease groups had comparable amounts of CD3(-)NCR1(+) cells in PBMC to the healthy individuals. All organs examined including liver, spleen and lymph nodes contained CD3(-)NCR1(+) cells. Circulating CD3(-)NCR1(+) cells were further characterized as CD56(-)GranzymeB(+)CD8(-). A CD3(+)NCR1(+) population was observed in PBMC in 79 % of the healthy dogs examined representing at the most 4.8 % of the lymphocyte population. In canine samples examined for CD56 expression, CD56(+) cells were all CD3(+) and NCR1(-). To our knowledge, this is the first examination of NCR1 expression in the dog. The study shows that this NK cell associated receptor is expressed both on populations of CD3(+) and CD3(-) blood lymphocytes in dogs and the receptor is found on a CD3(+) GranzymeB(+) CD8(+) leukemia. Our results support that CD56 is expressed only on CD3(+) cells in dogs and shows that NCR1 defines a different CD3(+) lymphocyte population than CD56(+)CD3(+) cells in this species. CD3(-)NCR1(+) cells may represent canine NK cells.

Listeria monocytogenes associated kerato-conjunctivitis in four horses in Norway.

Listeria monocytogenes has been reported to cause various infectious diseases in both humans and animals. More rarely, ocular infections have been reported. To our knowledge, only two cases of Listeria keratitis have been described in horses. We report kerato-conjunctivitis in four Norwegian horses associated with L. monocytogenes. Clinically, all cases were presented with recurrent unilateral kerato-conjunctivitis. L. monocytogenes bacteria were isolated from swab samples from all cases, and cytology carried out in 3 cases was indicative of L. monocytogenes infection. The present report describes the first known cases in which L. monocytogenes has been isolated from keratitic lesions in horses in Norway. A potential risk factor may be feeding of silage or haylage, but other sources of infection cannot be ruled out. The phenotypic features including antimicrobial susceptibility and serotype of the isolates are described. Laboratory detection of L. monocytogenes demands extra caution since only low numbers of bacteria were detected in the eye-swabs, probably due to the low volume of sample material and the intracellular niche of the bacterium. A general poor response to treatment in all these cases indicates that clinicians should pay extra attention to intensity and duration of treatment if L. monocytogenes is identified in connection with equine kerato-conjunctivitis.

Hematological shift in goat kids naturally devoid of prion protein.

The physiological role of the cellular prion protein (PrP(C)) is incompletely understood. The expression of PrP(C) in hematopoietic stem cells and immune cells suggests a role in the development of these cells, and in PrP(C) knockout animals altered immune cell proliferation and phagocytic function have been observed. Recently, a spontaneous nonsense mutation at codon 32 in the PRNP gene in goats of the Norwegian Dairy breed was discovered, rendering homozygous animals devoid of PrP(C). Here we report hematological and immunological analyses of homozygous goat kids lacking PrP(C) (PRNP(Ter/Ter) ) compared to heterozygous (PRNP (+/Ter)) and normal (PRNP (+/+)) kids. Levels of cell surface PrP(C) and PRNP mRNA in peripheral blood mononuclear cells (PBMCs) correlated well and were very low in PRNP (Ter/Ter), intermediate in PRNP (+/Ter) and high in PRNP (+/+) kids. The PRNP (Ter/Ter) animals had a shift in blood cell composition with an elevated number of red blood cells (RBCs) and a tendency toward a smaller mean RBC volume (P = 0.08) and an increased number of neutrophils (P = 0.068), all values within the reference ranges. Morphological investigations of blood smears and bone marrow imprints did not reveal irregularities. Studies of relative composition of PBMCs, phagocytic ability of monocytes and T-cell proliferation revealed no significant differences between the genotypes. Our data suggest that PrP(C) has a role in bone marrow physiology and warrant further studies of PrP(C) in erythroid and immune cell progenitors as well as differentiated effector cells also under stressful conditions. Altogether, this genetically unmanipulated PrP(C)-free animal model represents a unique opportunity to unveil the enigmatic physiology and function of PrP(C).

Dirofilaria repens infection in a dog imported to Norway.

Dirofilaria repens infection was diagnosed in a dog that had been imported to Norway from Hungary three years previously. The dog was a four-year-old castrated male mixed-breed dog and presented for examination of two masses on the right thoracic wall. Fine needle sampling from the subcutaneous nodules and subsequent cytological examination revealed a high number of microfilariae and a pyogranulomatous inflammation. At re-examination approximately 3 weeks later, both masses had apparently disappeared spontaneously, based on both inspection and palpation. However, examination of peripheral blood by a modified Knott's test revealed a high number of unsheathed microfilariae with mean length of 360 µm and mean width of 6-7 µm, often with the classic umbrella handle appearance of D. repens. Polymerase chain reaction and sequencing confirmed the D. repens diagnosis. Subcutaneous dirofilariosis caused by D. repens is probably the most common cause of human zoonotic dirofilariosis in Europe, but currently is rarely encountered in northern countries such as Norway. However, travelling, import and relocation of dogs have increased, and thus the geographical range of these parasites is likely to increase from traditionally endemic southern regions. Increasing numbers of autochthonous cases of D. repens infections in dogs have been reported in eastern and central Europe. Although infection with D. repens often induces only mild signs or subclinical infections in dogs, they nevertheless represent a reservoir for zoonotic transmission and thus a public health concern, and, in addition, due to the long prepatent period and the high frequency of subclinical infections or infections with unspecific clinical signs, could easily be missed. Lack of experience and expectation of these parasites may mean that infection is underdiagnosed in veterinary clinics in northern countries. Also, predicted climate changes suggest that conditions in some countries where this infection is currently not endemic are likely to become more suitable for development in the intermediate host, and thus the establishment of the infection in new areas.

Climate and environmental change drives Ixodes ricinus geographical expansion at the northern range margin.

BACKGROUND: Global environmental change is causing spatial and temporal shifts in the distribution of species and the associated diseases of humans, domesticated animals and wildlife. In the on-going debate on the influence of climate change on vectors and vector-borne diseases, there is a lack of a comprehensive interdisciplinary multi-factorial approach utilizing high quality spatial and temporal data. METHODS: We explored biotic and abiotic factors associated with the latitudinal and altitudinal shifts in the distribution of Ixodes ricinus observed during the last three decades in Norway using antibodies against Anaplasma phagocytophilum in sheep as indicators for tick presence. Samples obtained from 2963 sheep from 90 farms in 3 ecologically different districts during 1978 - 2008 were analysed. We modelled the presence of antibodies against A. phagocytophilum to climatic-, environmental and demographic variables, and abundance of wild cervids and domestic animals, using mixed effect logistic regressions. RESULTS: Significant predictors were large diurnal fluctuations in ground surface temperature, spring precipitation, duration of snow cover, abundance of red deer and farm animals and bush encroachment/ecotones. The length of the growth season, mean temperature and the abundance of roe deer were not significant in the model. CONCLUSIONS: Our results highlight the need to consider climatic variables year-round to disentangle important seasonal variation, climatic threshold changes, climate variability and to consider the broader environmental change, including abiotic and biotic factors. The results offer novel insight in how tick and tick-borne disease distribution might be modified by future climate and environmental change.

Multi-source analysis reveals latitudinal and altitudinal shifts in range of Ixodes ricinus at its northern distribution limit.

BACKGROUND: There is increasing evidence for a latitudinal and altitudinal shift in the distribution range of Ixodes ricinus. The reported incidence of tick-borne disease in humans is on the rise in many European countries and has raised political concern and attracted media attention. It is disputed which factors are responsible for these trends, though many ascribe shifts in distribution range to climate changes. Any possible climate effect would be most easily noticeable close to the tick's geographical distribution limits. In Norway- being the northern limit of this species in Europe- no documentation of changes in range has been published. The objectives of this study were to describe the distribution of I. ricinus in Norway and to evaluate if any range shifts have occurred relative to historical descriptions. METHODS: Multiple data sources - such as tick-sighting reports from veterinarians, hunters, and the general public - and surveillance of human and animal tick-borne diseases were compared to describe the present distribution of I. ricinus in Norway. Correlation between data sources and visual comparison of maps revealed spatial consistency. In order to identify the main spatial pattern of tick abundance, a principal component analysis (PCA) was used to obtain a weighted mean of four data sources. The weighted mean explained 67% of the variation of the data sources covering Norway's 430 municipalities and was used to depict the present distribution of I. ricinus. To evaluate if any geographical range shift has occurred in recent decades, the present distribution was compared to historical data from 1943 and 1983. RESULTS: Tick-borne disease and/or observations of I. ricinus was reported in municipalities up to an altitude of 583 metres above sea level (MASL) and is now present in coastal municipalities north to approximately 69°N. CONCLUSION: I. ricinus is currently found further north and at higher altitudes than described in historical records. The approach used in this study, a multi-source analysis, proved useful to assess alterations in tick distribution.

First case of babesiosis caused by Babesia canis canis in a dog from Norway.

An Irish setter from the Oslo area was presented to the clinic with signs of babesiosis, a few days after a tick bite. Blood analysis confirmed babesiosis. Microscopic examination of thin blood film revealed large, basophilic, bodies inside erythrocytes, indicative of a large Babesia sp. Molecular analysis using PCR, indicated the presence of a Babesia spp. in the blood. Sequencing and phylogenetic analysis of the PCR fragment revealed a sequence which was 100% identical to Babesia canis canis 18S. As this dog had never been abroad, it can be concluded that this is the first report of an autochthonous infection of B. canis canis in Norway.

Comparison between microscopic and automated differential leukocyte counts in the silver fox (Vulpes vulpes) and the blue fox (Alopex lagopus).

Differential leukocyte (WBC) counts in blood from clinically healthy silver foxes (n=32) and blue foxes (n=37) obtained from an automated hematology analyzer (Technicon H*1 Hematology System) with canine software were compared with microscopic differential WBC counts (M-diff). There was good agreement between the automated differential cell count (A-diff) and the M-diff for neutrophil and lymphocyte percentages. The correlation was lower for monocyte percentages and variable for eosinophil percentages. There was no significant difference between the A-diff and M-diff in either fox species. The A-diff counts were very precise, and may be a good alternative to the traditional M-diff for screening populations of clinically healthy foxes or for studies on stress and animal welfare. Intercept values, however, indicated a constant bias that must be taken into account before interpreting results based on different methods of analysis