In humans, genes coding for theα2-adrenoceptors are polymorphic resultingin variationsin the pharmacodynamic response to α2-agonists. Our aim was to assess the presence of polymorphismsin the gene encoding the α2A-adrenoceptor in dogs.
Dogswhichpresented to the veterinary schoolhospital forelectivesurgical procedures were enrolled. Ethical approvalwas obtainedandinformed consent was provided bythe owners. The waste blood from pre-anaesthetic bloodsampleswas retained for DNA extraction and geneticanalyses. The α2A-adrenoceptorwas amplified using the polymerase chain reaction (PCR)in two parts and sequenced with Sanger sequencing.The prevalence of polymorphismswas determined.
A total of 72 dogsof 43breeds, 15 crossbreds and oneof an unknown breedwere included in the study. Median body weight (range) was 15.6 (2.4–50.3) kg, with 32males and 40 females. Samples from five dogs could not be amplified. Only three polymorphic sitesin the gene were present, one synonymous transition as a heterozygote in the coding region in two dogs, one transition as a heterozygote in the non-coding regionin one dog, and one homozygous 22 base pairdeletion in the non-coding region in one dog. The overall nucleotide diversity was only 0.00004.
In this group of dogsof various breeds, nopolymorphismsaffecting the protein structure of the α2A-adrenoceptor were detected. However, a very GC-rich section of the gene could not be analysed. Further work is neededto determine whether this part of the genemay alter the response to administrationof α2-adrenoceptor agonists in dogs.
An increased incidence of bacterial pneumonia (BP) has been reported in Irish Wolfhounds (IWHs), and it is typical for the disease to recur repeatedly. The aetiology remains unclear, but the development of bronchiectasis in the absence of clinical findings indicative of interstitial lung disease is common in affected IWHs. This observation led us to the hypothesis that the root cause could be at the bronchial level.
This study describes the post-mortem histopathological changes of the lungs and bronchi and evaluates bronchial cartilage structure and composition in IWHs (n = 10) with previous pneumonias compared with control IWHs (n = 5) and other large breed control dogs (n = 5) without a history of respiratory disease. Histological evaluation of lung samples was performed with haematoxylin-eosin (HE), Masson’s trichrome and Prussian Blue staining as well as with immunohistochemical staining (IHC) for CD3, CD79a antigens and pan-cytokeratin (CK). Bronchial cartilage proteoglycan content was evaluated in safranin ‘O’ (SO) staining and chondrocyte apoptosis by IHC to cleaved caspase-3. Image analysis was used to measure the bronchial cartilage to lumen area ratio as well as the SO staining intensity and caspase-3 positive cell count in digital whole slide images.
According to this study, the most consistent histological finding in the lungs of IWHs with recurrent BP is a chronic lymphoplasmacytic inflammation in the peribronchial and perivascular pulmonary interstitium, along with remodelling of the bronchial walls. The bronchial cartilage to lumen area ratio was significantly lower in all IWHs than in control dogs of other breeds (P = 0.011), suggesting less cartilage support in the bronchi of the IWH breed. The SO intensity of the bronchial cartilage samples was significantly higher (P = 0.008) in affected IWHs than in control dogs of other breeds. The caspase-3 staining of large bronchial cartilages showed significantly less apoptotic chondrocytes in affected IWHs than in all control dogs (P = 0.045).
Based on these findings, altered cartilage metabolism, as a breed-related feature or secondary to chronic inflammation, may play a role in the pathogenesis of bronchiectasis development and recurrent lung infections in this breed.
Mycobacteriosis is a potential zoonotic disease routinely diagnosed in histology by Ziehl Neelsen (ZN) staining. Long standing antibiotic treatment can cause failure of the ZN staining to reveal the mycobacteria due to a disrupted bacterial capsule. This poster describes a canine case of pulmonary mycobacteriosis with loss of acid-fast bacterial properties.
A 3-year-old intact border collie bitch with severe lobar pneumonia, non-responsive to antibiotic treatment of several weeks duration was referred to the University of Helsinki Veterinary Teaching Hospital in June 2023 and underwent surgical lobectomy of the middle right lung lobe. The lobe was sent for pathological examination to identify the cause of the pneumonia. HE, ZN, modified ZN; Grocott's methenamine silver stain (GMS) and in-situ hybridization (ISH) with probe detecting bacterial 16S rRNA were performed. Microbiological culture was performed from the lung biopsy, bronchoalveolar lavage (BAL) fluid and thoracic cavity effusion as well.
HE staining revealed severe confluent chronic pyogranulomatous pneumonia in the resected lung lobe. ZN and modified ZN staining were negative in all sections. GMS revealed bacillus-like structures within the granulomas and chromogenic ISH confirmed these to be bacteria. Bacterial growth was seen in the BAL fluid after six days incubation and in the biopsy and pleuran effusion after two weeks incubation. The cultured bacterium was identified as Mycobacterium thermoresistibile by mass spectrometry.
This case highlights the importance of molecular methods for detecting bacterial infection, in addition to standard histochemical stains, in cases with a history of prolonged antibiotic treatment and a persistent tissue lesion suggestive of infection. This case also emphasizes the importance of communication between practitioners and pathologist.
Approximately third of agility dogs sustain sport-related injuries. We aimed to evaluate the effects of jump height on the landing kinetics.
Fifteen Border Collies (wither height 44-56 cm) performed bar jumps in a straight-line at 80%, 100% and 120% of the dogs’ height. Impulses of fore- and hindlimb pairs were calculated from forces measured by two force plates (AMTI). Trunk horizontal velocity at touch-down of trailing forelimb, landing distance and landing angle were calculated from reflective markers (Vicon Nexus, 13 cameras). Mean values were calculated from ≥2 trials/jump height. Repeated measures ANOVA was used for statistical analysis.
Trunk horizontal velocity at touch-down of trailing forelimb decreased with increasing jump height (p<0.001), landing angle became steeper (p<0.001), and landing distance increased (p<0.001). Mean differences between heights 80% and 120% were -0.86 m/s in horizontal velocity, 7.20 degrees in landing angle, and 0.58 m in landing distance. As jump height increased, the vertical impulse increased in forelimb and hindlimb pairs (p<0.001). Mean differences in vertical impulse between heights 80% and 120% were 0.42 Ns/kg in forelimbs and 0.17 Ns/kg in hindlimbs. The decelerative horizontal impulse was not affected by jump height in either limb pair (p>0.26). The accelerative horizontal impulse increased with jump height in forelimb pair (p<0.001) and hindlimb pair (p=0.002) with mean difference between heights 80% and 120% being 0.092 Ns/kg in forelimbs and 0.067 Ns/kg in hindlimbs.
Increased horizontal propulsion and vertical impulses of both limb pairs at higher jump heights may have an impact on sport-related injuries.
Administering medications to cats at home often poses challenges for owners, highlighting the importance of easy product administration in feline medicine. In two separate clinical field trials, we evaluated the usability of two different pregabalin formulations. These studies involved client-owned cats with a history of anxiety and fear associated with veterinary visits and/or travelling.
The first pilot study employed a cross-over design and used the pregabalin 20 mg/ml oral solution formulation (Lyrica®) registered for human use. This study involved administering doses of 5 and 10 mg/kg to 11 cats, with a total of 22 administrations. In the second study, a more concentrated pregabalin 50 mg/ml oral solution formulation (Bonqat®), specifically developed and flavoured for cats, was administered once at the label dose of 5 mg/kg to 108 cats. Owners evaluated the usability of both formulations using a 4-point rating scale (very easy, easy, somewhat difficult, very difficult), and the results were evaluated statistically.
Most owners found administering the human-registered formulation to be somewhat difficult (68%) or very difficult (5%), while some assessed it to be easy (14%) or very easy (14%). The volume of the treatment administered varied from 0.9 to 3.5 ml per administration. In contrast, the majority of owners assessed the administration of the cat-specific formulation as very easy (48%) or easy (31%), and only some as somewhat difficult (17%) or very difficult (5%). The range of dose volumes for this formulation was between 0.2 and 0.8 ml per cat. The difference between usability assessments of the two formulations was statistically significant (p<0.0001).
In conclusion, the novel feline-specific oral formulation was found to be easier to administer compared to the formulation registered for human use. The good usability of the cat-specific product can likely be attributed to its small dosing volume (0.1 ml/kg) and flavoured liquid formulation.
Brachycephalic obstructive airway syndrome (BOAS) is major welfare issues concerning many flat-faced dog breeds. Signs include respiratory difficulties, sleep disordered breathing and exercise and heat intolerance. Although various breeding schemes against BOAS has been implemented in recent years, their impact on breed health remains unknown. The aim of the current study was to evaluate Finnish Kennel Club (FKC) BOAS screening test results and estimate heritability of veterinary assessed BOAS grade.
FKC BOAS screening test includes 1000m exercise test with recovery and body temperature assessment, clinical BOAS grading that evaluates severity of upper respiratory signs before and after exercise, and evaluation of nostril stenosis. BOAS grade and level of nostril stenosis are recorded using four grade scale (no, mild, moderate, severe). Heritability analysis for BOAS grade was done by using parent–offspring regression model.
The majority (80%) of dogs (n=957) participating in FKC BOAS screening test were English Bulldogs, French Bulldogs, and Pugs. Percentage of their litters that had at least one parent BOAS tested increased from 17-34% in 2018 to 89-100% in 2022. The exercise test was most frequently failed by English Bulldogs (11%), followed by French Bulldogs (4%) and Pugs (3%). None of the other breeds tested failed the exercise part. Altogether 77% of tested dogs were BOAS- (i.e. had no or mild signs), but less than half of the dogs had open or only mildly stenotic nostrils. Proportion of moderate to severe nostril stenosis was highest (71%) in Pugs. Estimates of heritability for BOAS grade were separately calculated for the three most common breeds and for all dogs, and the estimates were moderate to high, ranging from 0.39 to 0.58.
Our results indicated that genetic improvement can be obtained, and severity of BOAS signs in offspring reduced by using clinical respiratory sign i.e. BOAS grading. Exercise test alone did not identify all dogs with moderate or severe BOAS signs. As BOAS is a complex disease and breed differences exist, it is advisable to evaluate exercise tolerance, thermoregulation, and nostril stenosis in addition to BOAS grading in all breeding animals.
Vatinoxan, an alpha2-adrenoceptor antagonist that poorly penetrates the blood-brain barrier, is used in veterinary medicine to attenuate the cardiovascular effects of medetomidine. The hemodynamic effects of the vatinoxan/medetomidine combination have not been previously studied in rats. We evaluated a combination of medetomidine (0.25 mg/kg) and midazolam (2 mg/kg; MM) with or without vatinoxan (5 mg/kg; MMV) in a randomized cross-over design using 10 adult male Wistar rats. We hypothesized that MMV would shorten the onset of sedation and provide reliable sedation with less hypertension and bradycardia compared with MM. Drugs were administered subcutaneously in the neck. Rats were maintained in a temperature-controlled chamber and provided with flow-by oxygen. The time to loss of the righting reflex was measured. Oscillometric mean arterial pressure (MAP) was monitored with a tail cuff for 55 minutes after injection. Pulse rate (PR) and oxygen saturation were monitored via pulse oximeter and recorded until 55 minutes after injection. All rats were administered atipamezole (1.25 mg/kg) and flumazenil (0.2 mg/kg) at the end of each trial. Compared to MM, time to loss of the righting reflex was significantly reduced (p=0.050) with MMV. PR was maintained significantly higher (p<0.05) and MAP significantly lower with vatinoxan (p<0.05), without causing hypotension (<60 mmHg). In conclusion, subcutaneous vatinoxan significantly shortened the onset of sedation and mitigated medetomidine-associated hypertension and bradycardia in Wistar rats administrated medetomidine and midazolam, showing potential for a safer and more efficient sedation of healthy, adult, male Wistar laboratory rats.
Rabbits kept under ultraviolet B (UVB)-irradiaNon respond with increasing serum total vitamin D levels. As UVB exposure increases also the ergocalciferol (vitamin D2) concentraNon in hay, it has been unknown whether irradiaNon of the animals or their feed contributes more to serum vitamin D concentraNons. Twenty-four female New Zealand White rabbits were divided into three groups for a four-week study period: the control group (C) received no UVB-exposure and non-irradiated hay (baseline ergocalciferol (vitamin D2) concentraNon of hay 2.05 μg/100 g dry maBer). The direct exposure group (D) was provided with 12 hours of UVB-irradiaNon daily and fed the same hay as group C in shaded areas to prevent UVB-irradiaNon thereof. The indirect exposure group (I) did not receive direct UVB-irradiaNon but was fed with hay that was similarly treated as the animals from group D (vitamin D2 concentraNon of hay 4.55 μg/100 g dry maBer). Serum 25(OH)D2, 25(OH)D3, ionised calcium, total calcium, phosphorus, and magnesium concentraNons were measured weekly. No systemaNc effect existed on serum mineral concentraNons. Serum 25(OH)D2 concentraNons were significantly higher in group I compared to groups C and D from the second week onwards. 25(OH)D3 levels increased only in group D, with significant differences to both other groups from the third week onwards, yet at lower magnitudes than the noted increase of 25(OH)D2 in group I. Total 25(OH)D levels were highest in group I, intermediate in group D and lowest in group C. Serum total 25(OH)D concentraNon was more affected by UVB-irradiaNon of rabbits’ feed than by direct irradiaNon of the animals. If rabbit serum total 25(OH)D levels should be managed, diet manipulaNon rather than animal UVB-exposure appears to be more effecNve.
Chronic inflammatory rhinitis (CIR) is among the most common causes of chronic nasal signs in dogs. The diagnosis of CIR is obtained by excluding other causes for chronic nasal signs, such as dental disease, aspergillosis and neoplasia, and histopathologic analysis of nasal biopsies. To our knowledge, there are no studies reporting the histopathological changes in the nasal mucosa of healthy dogs. Since the nasal cavity is in contact with inhaled microbes and particles, it is indicated to compare the histopathological findings in dogs without nasal signs to those with CIR to better understand the clinical relevance of inflammatory changes.
The purpose of this study was to describe the histopathological findings in the nasal biopsies of dogs without nasal signs compared to dogs with CIR. Study groups included control dogs without nasal signs euthanized for reasons unrelated to this study and donated for research purposes (n = 20) and previously collected archived nasal biopsies from dogs diagnosed with CIR (n = 20). The diagnosis of CIR was based on typical clinical presentation and computed tomography, rhinoscopy and histopathology findings indicative of CIR.
Nasal biopsy specimens were stained with H&E, Masson, PAS, Gram and evaluated with light microscopy. Inflammatory cell count was calculated for three random high-power fields (2.37 mm2). The number of lymphocytes and plasma cells was increased (P <0.001) in dogs with CIR (median 92, range 11-658) when compared to control dogs (median 19, range 1-57). The number of neutrophilic granulocytes (median 9, range 1-48; median 1, range 0-3; P <0.001) was increased in dogs with CIR. The number intraepithelial leukocytes (median 9, range 0-96; median 1, range 0-3; P <0.001) was increased and the epithelium was thicker in dogs with CIR. On the contrary, the number of visible goblet cells was decreased in dogs with CIR.
In conclusion, there is a small population of inflammatory cells in the nasal biopsies also in dogs without nasal signs. Many factors are altered in chronic inflammatory rhinitis, including the number of inflammatory cells, and epithelial alterations that are likely to play a role in the pathophysiology of the disease.
Medetomidine is a common sedative drug in dogs, and it can be combined with vatinoxan to reduce its cardiovascular effects. However, safety and efficacy of either medetomidine or medetomidine-vatinoxan has not been evaluated in puppies.
Healthy puppies aged between 6 – 8 weeks scheduled for elective brainstem auditory-evoked response (BAER) testing were recruited for the study. Puppies were randomised within each litter to receive either intramuscular medetomidine 10 μg kg-1 and butorphanol 0.2 mg kg-1(group MB), or medetomidine 10 μg kg-1, vatinoxan 0.2 mg kg-1 and butorphanol 0.2 mg kg-1(group MVB). Heart rate (HR) and non-invasive MAP were measured at baseline (BL) and 5 minute interval after sedation. Paired samples Student t-test was used to compare means.
All the puppies (n = 32 were sufficiently sedated for BAER testing within 5 minutes after injection. At BL, HR was 178 ± 14 and 172 ± 20 bpm (p = 0.335), and MAP was 104 ± 25 and 104 ± 18 mmHg (p = 0.973) in group MB and MVB, respectively. Between 5 – 15 minutes, HR was significantly higher in group MVB (142 ± 28, 154 ± 27 and 157 ± 17 bpm) than in group MB (109 15, 113 14 and 119 13 bpm) (p < 0.001). Whereas MAP was significantly lower in group MBV (99 8, 82 7 and 77 7 mmHg)than in group MB (111 12, 97 9 and 91 7 mmHg) (p < 0.001). No hypotension (MAP < 65 mmHg) was detected in any animal. All the puppies recovered from sedation uneventfully.
Medetomidine-butorphanol, with or without vatinoxan, is safe and effective sedative combination for puppies. The combination of medetomidine-butorphanol and vatinoxan maintains heart rate closer to baseline value.
In this study the current prevalence of endoparasites in Finnish dogs and cats was examined. Data from studied animals was collected for risk factors linked to Toxocara infections. Fecal samples from 664 dogs and 379 cats (including 46 shelter cats), over six months old, were examined with quantitative flotation method (Mini-Flotac®) using mainly zinc sulphate flotation solution, and magnesium sulphate for some samples. Fecal samples from 396 dogs and 89 cats were also examined with Baermann method in order to detect nematode larvae.
The overall prevalence of endoparasites was 3.5% in dogs, 3.6% in pet cats and 41.3% in shelter cats. Strongylids (1.7%; probably pseudoparasites due to coprophagy) were most common findings in dogs followed by Toxocara (0.9%), trematodes (0.4%), Eucoleus spp. (0.3%), and Taenia spp., Trichuris vulpis, Isospora spp. and Crenosoma vulpis (0.2% each). Toxocara was the most prevalent finding in pet cats (3.3%) and shelter cats (34.8%). Taenia (0.6%) and trematode (0.3%) eggs were also found in pet cats. Shelter cats were also found to be infected with Eucoleus spp. (13.0%), Isospora spp. (10.9%), Taenia spp. (8.7%) and Toxoplasma gondii/Hammondia hammondii (2.2%).
Dogs that were most likely infected with Toxocara were under twelve months old. In cats, shelter background, male gender, being mixed-breed, being dewormed two to four times a year and having history of previous parasitic infections was associated with higher Toxocara infection prevalence. The background data collected from the questionnaire revealed that 31.2% of studied dogs were dewormed or their fecal sample was analyzed less than once a year or never, which are both unacceptable, even though the prevalence is low.
Endoparasitic prevalence in Finnish dogs and cats has reduced since past studies and is currently very low. The Eucoleus prevalence in shelter cats was unexpectedly high.
Considering that dogs originate from carnivorous wolves; it may be questioned whether high amounts of carbohydrates are beneficial to their health. Low-carbohydrate raw meat-based diets (RMBDs) are increasing in popularity, but it is unknown how they affect dogs’ energy metabolism in comparison to a traditional high-carbohydrate kibble diet. We aimed to report the changes in energy metabolism biomarkers and bodyweight in dogs that ate either a kibble diet or a RMBD.
A diet intervention study including 46 client-owned Staffordshire Bull Terriers was performed. The dogs were fed either a high-carbohydrate kibble diet, or a low-carbohydrate RMBD for 50-188 days (median 136 days or 4.5 months). Before and after the trial, fasting blood samples were taken for analysis of glucose, glycosylated hemoglobin (HbA1c), insulin, glucagon, triglycerides, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), and 3-hydroxybutyrate (BHB). In addition, the homeostatic model assessment of insulin resistance (HOMA-IR) and the triglyceride glucose (TyG) index were calculated and analyzed, and bodyweight was evaluated. Comparisons were made between the two diet groups at baseline and end, and between baseline and end within both diet groups.
Feeding a kibble diet to pet dogs for 4.5 months significantly increased HbA1c (p=0.026), cholesterol (p<0.001), BHB (p=0.018), and bodyweight (p=0.018). Feeding a RMBD significantly decreased glucose (p=0.026), glucagon (p=0.004), cholesterol (p=0.027), and TyG index (p=0.033), and increased BHB (p<0.001). At the end of the trial, the kibble fed dogs had significantly higher levels of triglycerides (p=0.006), total cholesterol (p<0.001), HDL (p<0.001), LDL (p=0.014), and VLDL (0.007) compared to the RMBD fed dogs, and the RMBD fed dogs had significantly lower glucagon (p=0.004) and TyG index (p=0.017), as well as higher BHB (p=0.005) compared to the kibble fed dogs. No significant changes in insulin and HOMA-IR were found.
Based on the results of this study it can be concluded that a kibble diet, and a RMBD, respectively, have different effects on energy metabolism biomarkers in dogs. More research is needed to understand if this affects dogs’ health.
Hyaluronan (HA) is a non-sulfated glycosaminoglycan, which is a major component of extracellular matrix. There is increasing evidence, that HA is involved in both protective and pathological responses in lung inflammation and repair. During lung injury, low-molecular- weight HA is produced and contrary to physiological HA, it exhibits pro-inflammatory properties. Deposition of HA has been documented in varying lung diseases in humans and in experimental animal models, but the role of HA in canine lung diseases is yet undiscovered. Our hypothesis was, that bronchoalveolar lavage fluid (BALF) HA expression is altered in canine inflammatory bronchial diseases.
Privately owned dogs diagnosed with chronic bronchitis (CB, n=12) and eosinophilic bronchopneumopathy (EBP n=12) as well as healthy control dogs (n=11) were included in this study. Diagnosis was based on blood work, thoracic imaging, bronchoscopy and BALF cytology and culture. HA was measured in BALF samples using a sandwich-like ELISA. HA concentrations between different groups were compared with independent samples Kruskal-Wallis test and correlations were assessed using Spearman’s correlation coefficients.
BALF HA was significantly (P<0.001) increased in dogs with EBP (median 42.9 ng/ml, interquartile range [IQR] 31.4-186.7 ng/ml) when compared to dogs with CB (median 6.2 ng/ml, IQR 3.4-16.6 ng/ml) or healthy dogs (median 3.9 ng/ml, IQR 3.2-5.6 ng/ml). Additionally, HA was positively correlated to both blood eosinophil count (P=0.002, r= 0.451) and BALF eosinophil percentage (P=0.001, r=0.471).
These results show that HA concentration in BALF increases in canine eosinophilic lung disease, similarly, as reported in human allergic asthma. This indicates that HA has a role in EBP pathophysiology and allows further studies investigating the potential of HA as a diagnostic biomarker or as a therapeutic target.
Periodontal disease is a common inflammatory condition caused by dental plaque accumulation. Traditional treatments often involve mechanical plaque removal and adjunctive synthetic antimicrobials, which can cause irritation and disbalance in the oral microbiota. The aim of this study was to evaluate in vitro the anti-inflammatory and anti-plaque efficacy of spruce resin extract and pinoresinol, a compound found at the highest concentration in spruce resin extract. In addition, biocompatibility of the spruce resin extract-based toothpaste was evaluated for use in oral care formulations.
The anti-inflammatory activity was assessed using U937 macrophage-like cells stimulated with 1 μg/mL lipopolysaccharide (LPS). Cells were treated with spruce resin extract and pinoresinol at concentrations of 1%, 5%, and 20%. Cytokine levels of IL-1β, TNF-α, and MMP-3 were quantified using multiplex bead-based Luminex™ technology after 24 hours. Consequently, a toothpaste formulation containing 20% spruce resin extract was tested for its ability to inhibit biofilm formation. Glass rods were incubated with human saliva and exposed to the test formulation. Plaque growing on the roughened glass rods was quantified by dry weight (g) and the total viable bacterial counts (CFU). Finally, the biocompatibility of the toothpaste was evaluated using human oral epithelium (HOE) and gingival epithelium (HGE) 3D models. The models were exposed to the formulation for 3 minutes and 1 hour. Cell viability was determined using the MTT assay.
Treatment with spruce resin extract at 20% concentration resulted in a significant reduction of IL-1β, TNF-α, and MMP-3 levels compared to the LPS-only control (p ≤ 0.05), with efficacy comparable to that of dexamethasone. However, a significant increase of IL-1β in 20% pinoresinol-treated cells was observed compared to 20% vehicle-treated cells, suggesting that the anti-inflammatory effect of the extract was not solely due to pinoresinol. The anti-plaque assay demonstrated a 73% reduction in plaque biofilm biomass and a 90% reduction in CFU counts with spruce resin extract-based toothpaste compared to untreated controls, with efficacy slightly lower than that of Corsodyl mouthwash (containing chlorhexidine digluconate 0.2 % w/v), which achieved an 82% reduction in plaque biofilm biomass and a 99% reduction in CFU counts. The spruce resin extract-based toothpaste was also significantly more effective compared to Virbac C.E.T. Enzymatic toothpaste which achieved a 24% reduction in plaque biofilm biomass and a 15 % reduction in CFU counts. Biocompatibility testing demonstrated that the 20% spruce resin extract toothpaste did not exhibit corrosive effects, with cell viability remaining at 76% in oral epithelium (HOE) and 61% in gingival epithelium (HGE) after 3 minutes of incubation (OECD guidelines require at least 50%), and 33% in HOE and 46% in HGE after 1 hour of incubation (OECD guidelines require at least 15%).
The findings confirm the effectiveness of spruce resin extract 20% as a potent anti-inflammatory and anti-plaque agent. The study also establishes the biocompatibility of the formulation, suggesting that the toothpaste is non-corrosive and safe for oral use. These results support the potential use of spruce resin extract in developing natural, biocompatible oral care products.
Human Noroviruses (HuNoVs), which are members of the Caliciviridae family, are a common cause of viral foodborne outbreaks. The main ingredient of smoothies are berries which can be a source of HuNoVs. Smoothies are well-liked due to their delicious and nourishing features. The aim of our study was to determine the persistence of murine norovirus (MuNoV) in berry smoothies with milk or without milk. Since HuNoV is difficult to cultivate in cell culture lab we have used MuNoVs. We have used raspberry and blueberry smoothies in our experiments on various time points and temperatures (0 h - 4°C, 1 h - 4°C, 1 h -RT, 1 h - 37°C, 24 h - 4°C, 24 h - RT, and 24 h - 37°C). MuNoVs (MNV-1) and RAW 264.7 cells were used in the experiments. The smoothie was made in a blender using blueberries or raspberries, semi-skimmed milk, and some banana. In one experiment, PBS was used in place of milk. Samples were prepared in 10-fold dilutions and added to the cells. TCID50 assay was used to assess the virus load and cytopathic effect of the cells was examined using microscopy. There was a decrease in the MuNoV infection in the raspberry smoothie at 37°C as compared to 4°C and 21°C. TCID50/ml values stayed mostly consistent about ~ 6 log10 at 4°C and 21°C but reduced to ~ 5.5 log10 at 37°C after 24h. A faster decline in MuNoVs was also observed when PBS was substituted for milk in the blueberry smoothie. If NoV-contaminated berries are used in the smoothies and the ingredients are not heat-treated, they could result in foodborne illnesses. As a first finding, we found that milk might shield MuNoVs in blueberry smoothie from inactivation.
Research into microbial metabolism is essential to understand spoilage. In this study, the quality of modified atmosphere packaged (40% CO2: 60% N2) broiler was investigated during storage at 4°C and 6°C. The products were assessed daily by sensory evaluation and the study was terminated on the day after the sensory panel rated the samples to be spoiled. In addition to sensory evaluation, bacterial counts were determined daily using selective/specific media for the major bacterial groups, and samples were taken for nontargeted metabolomics analysis (ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry, UPLC-QTOF-MS). Spoilage was detected by sensory evaluation on day 8 at 6°C and three days later (day 11) at 4°C. The odor of the spoiled broiler was described as a sour. Also sulphur odor was observed in broiler stored at 6°C. Total bacterial levels reached log6 on days 7 and 5 and the log8 on days 12 and 7 at storage temperatures of 4°C and 6°C, respectively. Lactic acid bacteria (LAB) dominating at both storage temperatures, while enterobacteria were present at notable levels (> log 4) only in the broiler stored at the higher storage temperature. A total of 31712 metabolites were identified, mostly small peptides, amino acids, and polyamines. Over half of the observed metabolite differences (Spearman correlation, q < 0.05) were related to sensory evaluation (18438) and bacterial growth (15616). Most of these features (14653) were common to both variables. A total of 10930 metabolites varied with time, while only 29 differences were related to temperature. In both temperature groups, the concentration of inosine-5'-monophosphate (IMP) decreased with time, reaching a very low level in samples stored at 6°C. This suggests microbial utilization of ribose, which is a common feature for meat LAB. Since storage temperature significantly affected the microbiota, we also expected a greater impact on the metabolite profile. Particularly interesting are the changes in amines. Among biogenic amines, the amount of tyramine increased, especially at 6°C. Spermidine was only formed at 6°C, and further research will investigate its relationship with bacterial growth. The volatile profile of the samples will be examined in future studies.
While most food business operators (FBOs) comply with the legislation, non-compliances are detected in 15% of food control inspections in Finland. Additionally, 6% of FBOs violate food safety legislation repeatedly. As these FBOs cause a persistent health risk for consumers, inspectors should have more effective control tools available.
The purpose of this study was to identify challenges in the control of FBOs repeatedly violating food safety legislation. Additionally, the aims were to understand how control measures should be developed and to identify the training needs of inspectors.
A questionnaire regarding the control of FBOs repeatedly violating food safety was sent to all inspectors in the local food control units (N=62) in Finland.
Answers were received from 95 inspectors from 30 (48%) food control units and most of the respondents (80%) considered FBOs repeatedly violating food safety legislation as a fairly or very big problem. The most common challenges reported by inspectors included lack of knowledge of the FBO regarding legislation (87% of respondents), food hygiene risks (86%), and differences in risk perception between FBO and inspector (72%). Respondents wanted to improve their skills in the use of both soft measures such as motivating FBOs (53%) and hard measures such as coercive measures (59%) to reduce non-compliances. Compared to inspectors with sufficient time, the respondents with insufficient time for inspection and guidance had more FBOs with repeated violations to inspect (p=0.03), more challenges with FBO’s negative attitudes (p=0.04) and wanted more training on coercive measures (p=0.02).
The results can be used to develop food control strategies, as this study shows that improving effectiveness of the control of FBOs repeatedly endangering food safety requires multiple improvements. Some inspectors need more time for inspections whereas others need training in the use of both soft and hard methods.
Yersinia pseudotuberculosis is a psychrotrophic, facultative anaerobic, foodborne pathogen, whose reservoirs and epidemiology are partially unknown. The bacterium has caused several yersiniosis outbreaks in Finland, especially via carrots, lettuce, and raw milk. The aim of our study was to analyse the population genetics of Finnish Y. pseudotuberculosis strains using a WGS approach. We studied 93 strains isolated between 1974 and 2022 originating from wild animals (n=27), food production animals (n=20), companion animals (n=18), and humans (n=12), as well as various niches in the food chain (n=16) including raw milk, carrots and lettuce processing. All strains were serogrouped in silico with BLAST. Sequence types (STs) and core genome sequence types (CTs) were determined using schemes of seven housekeeping genes (Achtman scheme) and 1,553 core genes (EnteroBase), respectively.
We found four serogroups, six STs and 72 CTs. The most frequent ST-serogroup combinations were ST42-O:1a (n=36), ST43-O:1b (n=20), and ST19-O:3 (n=11). The virulence gene profiles were related with the serotypes and STs. Y. pseudotuberculosis strains isolated from humans had same STs with some strains of wild, companion and production animal origin as well as strains originating from raw milk industry. According to the CTs, six human strains had differences of six (0.37%), eight (0.52%), ten (0.64%), 11 (0.71%), 12 (0.77%) and 12 (0.77%) alleles with sporadic strains isolated from a guinea pig, hare, wild bird, guinea pig, cat, and hare, respectively. Production, companion and wild animals shared highly related CTs. Most pig strains (77%) were ST19-O:3 or ST583-O:3 and belonged to a pig-specific group with only one cat strain but no human or wildlife strains. However, some ST42 and ST43 strains were isolated both from pigs and humans.
In conclusion, highly related Y. pseudotuberculosis strains are found in humans, wild animals, companion animals and production animals. Hence, various animals can be sources of Y. pseudotuberculosis for humans. However, more studies are needed to understand the population genetics of strains isolated from humans and the emergence of Y. pseudotuberculosis outbreaks.
The rise of antimicrobial resistant bacteria is a global One Health threat with serious health risks if not addressed accordingly. Especially carbapenemase-producing Enterobacterales can cause life-threatening infections. One potential way to monitor the levels of antimicrobial resistant bacteria is by wastewater-based surveillance.
The aim of this study was to characterize multidrug-resistant bacteria isolated from wastewater influx samples taken approximately bi-monthly from February 2022 to March 2023 from ten different Finnish wastewater treatment plants (WWTPs). Samples were taken as part of Wastewater-based Surveillance as Pandemic Preparedness Tool (WastPan) consortium project. Bacteria was isolated by selective culturing (CHROMagar mSuperCarba and MRSA), isolates were identified using MALDI-TOF MS and DNA was extracted prior to this study. A total of 186 presumptive carbapenemase-producing isolates, Acinetobacter baumannii (n=8), Citrobacter freundii (n=92), Klebsiella pneumoniae (n=52), Escherichia coli (n=29) and Enterobacter cloacae (n=5), were screened for five of the most common carbapenemase-encoding genes blaKPC, blaNDM, blaOXA-48-like, blaVIM and blaIMP by PCR from the extracted DNA. The spa types of presumptive methicillin-resistant Staphylococcus aureus (MRSA) isolates (n=7) were determined by Sanger sequencing of the spa gene fragments amplified by PCR.
For the presumptive carbapenemase-producing Enterobacterales, a total of 68% of isolates (126 of 186) carried some of the tested carbapenemase-encoding genes. None of the isolates were positive for more than one resistance gene simultaneously. Among the five screened genes, blaKPC was the most prevalent at 41% (n=76), followed by blaOXA-48-like and blaNDM, at 15% (n=27) and 10% (n=19), respectively. Four isolates were positive for blaVIM and none for blaIMP. In total, 60 isolates (32%) tested negative for all carbapenemase genes. Statistical analysis (Kruskal-Wallis test) indicated no significant differences in carbapenemase-gene prevalence across the ten WWTPs (p > 0.05). Detected presumptive MRSA spa types were t304 (n=2), t026 (n=2), t002 (n=2) and t1339 (n=1). In conclusion, municipal wastewater contains a plethora of multidrug-resistant pathogens, especially C. freundii and K. pneumoniae expressing blaKPC. blaKPC is the most prevalent carbapenemase-encoding gene family in clinical samples and MRSA spa-type t304 has a clinical significance in Finland which both seems to be reflected to wastewater. However, these isolates should be further studied with whole-genome sequencing to further understand the genomic epidemiology of carbapenemase-producing Enterobacterales and MRSA. In general, wastewater surveillance has potential in antimicrobial resistance surveillance. Nevertheless, to gain further insight into these isolated bacteria and their antimicrobial resistance profiles, more in-depth analysis should be completed.
Food control inspections are an essential control measure to ensure compliance, and it is important to understand possible temporal patterns of inspection outcomes to develop food control. Seasonality of inspection outcomes and reasons for it are poorly known and need to be further investigated.
The aim of this study was to assess possible seasonal patterns in the compliance level of food service establishments in Finland, and to evaluate potential reasons explaining them.
We analysed inspection reports (n=86 223) of food service establishments in Finland from the years 2016–2021. Seasonal patterns of the overall inspection grades were analysed with monthly seasonal index analysis. Furthermore, we compared inspection grades of individual inspection items (n=37) between summer and the other time of the year with the chi-squared/Fisher’s exact test. Our analysis showed that the monthly seasonal indices were highest in June–August (1.13–1.17) and lowest in October–May (0.92–0.96) meaning that non-compliances were most common in the summer months. Establishments that operate only in summer did not explain the differences.
Furthermore, significant differences (p<0.05) in inspection grades were seen in 23/37 items, which all had inferior grades in summer compared to the other time. The items were related to a wide range of topics, some possibly related to warmer ambient temperature during summer (e.g., temperature management and vermin control), and some likely not, such as monitoring employees' health status, verification of hygiene proficiency, and hand hygiene.
Non-compliances were revealed to increase in summer in multiple items of which many have been associated with an increased risk of foodborne illnesses. The seasonality of non-compliances concerns year-round operating food service establishments, and it should be acknowledged by food business operators and also considered by inspectors in the timing of inspections.
Thermocautery disbudding is a common practice in dairy husbandry worldwide performed primarily for safety purposes. However, it elicits severe acute pain in calves resulting in noticeable behavioral and physiological responses. Hence, effective pain alleviation is crucial to mitigate suffering and promote animal welfare. This study aimed to explore the effects of levomethadone and ketamine on post-disbudding pain and activity in calves compared to xylazine alone.
Twenty-eight dairy calves aged 21 ± 5 days and weight 61.2 ± 9.4 kg were randomly assigned to receive three different sedation regimens as follows: Group XL (n = 10), levomethadone (0.05 mg kg-1) and xylazine (0.1 mg kg-1); group XK (n = 9), ketamine (1 mg kg-1) and xylazine (0.1 mg kg-1); and group X (n = 9), xylazine (0.2 mg kg-1) given intramuscularly 30 minutes before disbudding. Cornual nerve blocks (procaine HCl, 4.5 mg kg-1) and NSAID (meloxicam, 0.5 mg kg-1) were subcutaneously administered 15 minutes after sedation. The pain was subjectively assessed with a multidimensional pain score (0–21; 0 indicating no pain and 21 indicating severe pain). Pressure pain thresholds around the horn bud were measured with algometry (8–80 Newtons) before sedation and at specific intervals thereafter. Additionally, calf activity was monitored using two accelerometers between midnight and 6 am before and after disbudding. Differences between groups were analyzed using the Kruskal-Wallis test followed by Bonferroni corrections, whereas paired T-tests were used to compare activities within each group.
Four hours post-disbudding, the pain score was significantly higher with XK compared to X and XL [14 (11–16) versus 10 (7–14) and 10 (8–12), p = 0.045 and 0.008, respectively]. Pressure pain thresholds were significantly higher with XL than with X and XK [17 (12–24) versus 8 (8–31) and 8 (8–15) Newtons, p = 0.01] 24 hours after disbudding. While no significant differences were observed in overall calf activity between treatment groups, resting duration was decreased (302 ± 20 versus 278 ± 18 min night-1, p = 0.02) and standing duration was increased (58 ± 20 versus 82 ± 18 min night-1, p = 0.02) in group X post-disbudding.
In conclusion, the combination of levomethadone and xylazine appeared more effective in alleviating post-disbudding pain in calves than xylazine alone or xylazine combined with ketamine.
On-farm medication of sows is common. Complex medication management and use process of farms may contribute to medication safety risks, referring to safety risks associated with the use of medicines (e.g., using unclean equipment). These are largely unstudied. Our aim was to investigate the risks associated with the medication management and use process of sows during medications. This comprised implementation of medications, storage, equipment use, instructions and recording at farm level.
A convenience sample of 14 Finnish sow herds were recruited to an observation study in spring 2023. Herd size ranged from 150-1150 sows. Representatives of sow farms signed an informed consent and completed an electronic pre-questionnaire. The lead author visited the herds once and collected the observation data together with the person responsible for sow medications using a semi-structured observation form. All data were analyzed using descriptive statistics.
The pre-questionnaire respondents were farm owners (86%) and workers (14%) responsible for sow medications. The workers were mainly the ones who started, administered, and monitored the treatments. According to the responses, 36 % of the farms had non-Finnish speaking workers. All farms (n=14) had only veterinary medicines in the refrigerator, the temperature of which was within the reference values on 10 farms (71%). Five farms (36%) monitored the refrigerator temperature daily and five farms (36%) weekly. Five farms (36%) marked the opening date to a medication package. Look-alike and sound-alike (LASA) medications were used in 14% of the farms. Automatic medication syringes were used in 11 farms (79%) out of which nine (81%) had a functional syringe washing facility. Three farms (27%) said that they regularly disinfect their syringes. Eight farms (57%) changed the needle between sick sows. Written medication instructions were available in the piggery office for workers on eight farms (57%) and complete medication records on 11 farms (79%).
Several medication safety risks were identified on the farms. Based on observations, there is room for improvement in several areas, including employee guidance, medication instructions, storage, equipment hygiene and record keeping. Larger-scale future studies are needed for risk identification and to develop systems-based strategies to proactively manage risks before harm occurs.
Castration and local anaesthetic injections are painful for piglets. Restraining pigletsfor these procedures further exacerbates stress. We compared the impact of two different restraining devices: tubular (TUBE) and hanging (HANGING), on piglets’ behavioural reactions, body temperatures, and vocalizations during handling, local anaesthetic injections, and castration.
Male piglets (179, aged under five days) were randomised to either TUBE orHANGING and to receive either local anaesthesia (LA, procaine with adrenaline) or sham handling (SHAM) before castration. Three observers scored piglets’ behavioural reactions (0=no reaction, 10=the worst reaction imaginable) at four time points: when the piglet was restrained for the first time (FIRST), during local anaesthetic injections or sham handling (LOCAL), during the second time piglet was confined (SECOND), and during castration (CASTRATION). The effects of the treatments were analysed using mixed models. Results regarding body temperatures and vocalizations are being analysed.
Piglets reacted more when restrained in the TUBE compared to the HANGING device (4.6 ± 0.3 for TUBE vs. 2.8 ± 0.3 for HANGING (p<0.001) at FIRST and 4.8 ± 0.3 for TUBE vs. 3.3 ± 0.3 for HANGING (p<0.001) at SECOND). Local anaesthesia elicited stronger reactions than sham handling (p<0.001, LA 5.4 ± 0.3 vs. SHAM 3.8 ± 0.3), but reduced piglets’ reactions during castration (p<0.001,LA 4.9 ± 0.2 vs. SHAM 8.1 ± 0.2).
The pain-alleviating effect of local anaesthesia was highlighted, as was the pain of local anaesthetic administration. The restraining method affects piglets’ reactions, potentially affecting the validity of the assessment of local anaesthesia efficacy.
When milk production starts high-energy feeding starts and cows become prone to stress and metabolic and infective diseases. Coping depends on the facilities, for example length of the feeding fence and group size. Cows high in hierarchy might do better in the battle of resources. This could potentially affect rumen pH, rumination time and milk production. The aim was to study the associations between hierarchy of dairy cows, rumen pH, rumination time and milk production during transition period until 100 DIM.
Before calving, we inserted boluses (SmaXtech animal care®, Austria) that measured pH of the reticulum and a rumination index using online technique for 20 cows. Automatic milking systems measured milk production and cow weight. Hierarchy of cows was determined in two occasions (first from 33 to 78 DIM and second from 94 to 139 DIM). At this point, 9 first calvers and 8 older cows remained in the herd. The observation time was 12 and 8 hours, using video recording as an aid. An index of success in competitive interactions was calculated for each cow by dividing the number of replacements by the total number of interactions (either being replaced from the feeding fence or replacing another cow from it). The cows were fed mixed-ration supplemented with concentrates up to 7 kg from the milking robot. Dry cows were fed total mixed ration consisting of grass silage and hay complemented with concentrates. We described associations between hierarchy, reticular pH, rumination, and milk production.
Eleven cows belonged to low-success group (LH) and 6 cows were higher in hierarchy (HH). This result correlated well with age; all first-calvers and 2 older cows were LH. Cows experienced a decline in reticular pH after calving. The pH was normalized by 10 DIM to 6,9 (HH) and 6,87 (LH) compared to 6,7 before calving. The nadir seemed lower for HH cows (mean 6,4 and mean of the lowest values at 2 DIM 6,16) compared to LH cows (6,6 and 6,3). After 15 DIM, milk yield of HH cows was 44,5 kg milk/day, achieving a peak in 25 and starting to decline after 50 DIM. LH cows achieved a stable level of 34,5 kg milk 3 weeks post partum. After calving, HH cows weighed 706 (632 – 787) kg and lost ̴10 kg during 30 days. LH cows weighed 586 (498 – 684) kg, and lost 12kg during 14 days. However, the increase of feed intake and milk yield partially mask weight change. There were no big differences in rumination time between the groups (both roughly 600 min/day), although the nadir reached 325 min for HH cows at 1 DIM, compared to 360 min in LH cows.
High level of fiber in feed and several feeding times keep rumen pH at optimal level and rumination time high. However, there is a risk of a short period of subacute rumen acidosis immediately after calving specially with dominant cows. They are older, heavier cows with higher milk yield and production peak than primiparous cows low in hierarchy.
The wooden breast (WB) myopathyaffecting the large broiler breast muscles (pectoralis major) is a difficult problem in the poultry industry.Albeit eating WB meat is not a human health risk, it poses a threat to animal welfare.Severe WB observed at slaughter is associated with an impairment of gait scores, pointing to probable pain or distress to the living bird. It also causes economical losses and increased environmental load due to valuable breast meat condemnation. Globally,the prevalence of severe WB at slaughter is 5-10 % or higher.
Macroscopically WB myopathy manifests as bilateral palpable focal cranial to diffuse hardness, swelling and paleness of the breast muscle. Fibrinousexudate and hemorrhagingon the muscle surface may be present in extreme cases. Histologic characteristicsof WBinclude multifocal polyphasic myodegeneration with interstitial connective tissue accumulation and/or fibrosisas well asvariableperivascular inflammatory cell infiltrates, altered myofiber architecture, fat deposition and abnormal thickening of perimysial connectivetissue.The myodegeneration typically starts as a local lesionin the cranial part of theP. major affecting principally the superficial muscle.
The etiopathogenesis of myodegeneration in the WB myopathy is still unknown but hypoxia and HIF-1αpathway in the muscle have been linked to the changes. We hypothesized that in WB myopathy vascular development lags the muscle fiber growth leading to inadequate muscle perfusion and hypoxia. In this study, blood flow and vascularization of the broiler breast muscle were studied employing repeated PET/CT imaging utilizing two radioactive tracers. Biopsiestaken after each scanareused to relateand complementthe imaging findings withhistological muscle lesions.
In our study altogether 10 birds were imagedand sampled under anesthesiain Turku PET center three times: at the age of 14, 28 and 42 days using a PET Vision device. For the imaging,oxygen-15 labelled water (radio water) and [68Ga]-NODAGA-RGD tracer were used. Radio water was administeredas an intravenous bolus and dynamic PET was performed over 6 minutes. Then the [68Ga]Ga-NODAGA-RGD was injected as an intravenous bolus followed by dynamic PET over 20 minutes. Images are analyzed using Carimas 2.10software.
Amuscle biopsywas takenunder anesthesiafrom theprimarily affected muscle site at the first two timepoints and after euthanasia at the last timepoint.Muscle sampleswereevaluated for WB lesions.A monoclonalαvß3antibody will be usedto evaluateRGD-binding to developing vasculature.IBA1antibody for evaluating the inflammatory cell infiltrates,HIF1αantibody to detect tissue hypoxiaandIGF1R antibody for evaluation of IGF1R expression in the muscle.
The PET/CT imaging using selected tracers turned out to be a viable method to evaluate muscle perfusion and vascularization in growing broilers. Combination ofthe imaging study andhistology will expectantly allow us to correlate the breast muscle blood circulation to the advancement of the histological lesions. Preliminary results indicatedifferences between the perfusion in differentpartsof the muscle.Initial histological evaluation displays varying severities of WB changes, which gives us a valuableinsight for the underlying vascular development in disease progression.
During 2012-2015, we performed a research project on infectious hoof diseases in Finland. In this project, we visited dairy herds with an acute outbreak of interdigital phlegmon (IP). On the herd visits, we sampled IP cows and from the bacteriological swab samples Fusobacterium necrophorum was cultured and detected by PCR. In 2021, a selection of these F. necrophorum isolates were further characterized using whole genome sequencing (WGS). We wanted to investigate if the Finnish F. necrophorum isolates were genetically similar and if a particular infectious strain was spreading between the herds.
In our research project, we visited altogether 19 commercial dairy herds with an outbreak of IP. In these herds, we focused on IP and investigated mainly cows in an acute stage of IP (acute IP), which was diagnosed if a symmetric swelling and possible ulceration appeared in the interdigital cleft. For WGS , we selected isolates from those herds where we had detected at least two acute IP cows and had isolated F. necrophorum from IP lesions. In all, we had 27 F. necrophorum isolates from 27 cows in 7 herds: 25 IP cows, 1 digital dermatitis and 1 white line abscess.
The isolates were subjected to WGS at the Finnish Food Authority. DNA was isolated using the DNeasy Blood and Tissue kit (Qiagen, Germany). Libraries were prepared using the Illumina DNA Prep® library preparation kit (Illumina, USA) and sequenced on MiSeq (Illumina, USA). Good quality raw reads were assembled using Velvet software. The schema for an ad hoc gene-by-gene analysis included 1931 target genes and was defined using the Ridom SeqSphere+ software version 8 (Ridom, Germany). The core genome multilocus sequence typing (cgMLST) analysis was performed and a minimum spanning tree (MST) was created for the visualization of the differences.
Our findings revealed distinct clustering of Finnish F. necrophorum isolates from the seven herds, with each cluster being specific to its respective herd. Within a single herd, allellic differences between the isolates varied within a range of 0 to 2. In contrast, allelic differences varied from 21 to 870 between the samples isolated from separate herds.
Contrary to the presence of a single highly pathogenic infectious strain, our study identified herd-specific strains of F. necrophorum among acute IP cases in Finnish dairy cattle herds. This suggests a localized distribution pattern rather than widespread transmission of a particular virulent strain. FutherWGS-based research on F. necrophorum from an IP outbreak herd, exploring variations within the herd and potential virulence genes, would be fascinating.
Crayfish plague is a lethal disease to most freshwater crayfish species, except for North American crayfish species, which function as natural carriers of the crayfish plague agent, the oomycete Aphanomyces astaci. The detectionof A. astaciused to be dependent on the isolation of the agent, followed by pathogenicity test, until molecular diagnostic methods became available. Today, quantitative real-time PCR (qPCR) methods are most used for the diagnosis of crayfish plague. However, new discoveries of Aphanomycesspecies, such as A. fennicus, have demonstrated that false positive results are possible with the established internal transcribed spacer (ITS) based qPCR.Several PCR methods have been developed for detection of A. astaci, some of which amplify genotype-specific regions. Isolation of the pathogen remains a necessity for further research, although it is best suited for samples from acute disease cases. We compared three PCR methods and developed a MALDI-TOF MSmethod for the quick identification of isolates from cultures.Selected samples from a crayfish plague positive group of noble crayfish were evaluated by three PCR methods: the establishedITS-based qPCR, an improved ITS-based qPCR, and a genotyping PCR method.
For the MALDI-TOF MS analysis, an A.astacireference strain was used to create a mass spectrum library against which A. astacistrains of several genotypes. Protein-rich eluent was extracted from liquid cultures using a slightly modified formic acid and acetonitrile extraction commonly used with filamentous fungi.
Severalother oomycete species in Finnish Food Authority culture collection were compared to assess the specificity of the method.
The results of the three PCR methods from same crayfish samples are presented and compared.Using MALDI-TOF MS pure cultures of different A. astaci genotypes were reliably identified as A. astacifrom other Aphanomyces species and several other oomycete species tested.
Although molecular methods have improved the diagnostics of crayfish plague, the possibility of unknown genetic diversity of oomycetes in aquatic environment can affect the reliability of genetic methods. A method best fit for purpose should be chosen. MALDI-TOF MS appeared suitable as an additional diagnostic tool for A. astaciidentification of isolated cultures, with the caveat that the culture needs to be as pure as possible.
Rainbow trout gastroenteritis (RTGE) has been reported in Finland since 2010. RTGE is summer enteric syndrome and has been repeatedly isolated especially in RAS systems from 0-1 year old rainbow trout and has serious effects on production. In microscopically and histologically Segmented Filamentous Bacteria (SFB) are observed in intestine of fish in association with RTGE. The SFB has not been able to be cultivated or isolated in traditional methods, which has hindered studies of its association as agent of RTGE. With molecular methods SFB has been included in “Candidatus arthromitus” bacterial group. The aim of this study was to confirm presence of C. arthromitus in microbiota in rainbow trout with RTGE by comparing it for healthy fish. Furthermore, studies of affect of fish diseases on fish microbiota are scarce and microbiota changes in intestine affected by RTGE was studied. All together 39 Rainbow trout were collected from several fish farms with and without RTGE symptoms. Pathological changes and SFB present in intestine was studied microscopically. Intestine samples were taken for DNA extraction and sequenced with NGS method using Illumina MiSeq with 16S V3-V4 primers. From rainbow trout with clinical symptoms of RTGE and SFB confirmed on intestine with microscopic observation, also C. arthromitus bacteria was confirmed abundantly in intestine. From healthy fish C. arthromitus was not sequenced. Microbiota of healthy fish intestine had abundantly bacteria from phyla proteobacteria, while bacteria from proteobacterium phylum were less in intestine of RTGE fish. Furthermore, Clostridia bacterial group was more abundant in RTGE fish than healthy fish. This study shows that RTGE infection changes microbiota of rainbow trout by abundant presence of C. arthromitus but also by reducing proteobacteria, which many are considered beneficial for healthy intestine microbiota of fish.
Jalkapohjamuutokset ovat munintakanoilla merkittävä hyvinvointiongelma. Jalkapohjamuutoksia ovat hyperkeratoosi eli sarveistuminen ja eriasteiset jalkapohjatulehdukset ja -paiseet. Kuivikkeen kosteus ja likaisuus altistavat jalkapohjatulehduksille ja orsien kosteus lisää jalkapohjatulehduksien vakavuutta.
Epäsopivat orsimuodot ja ritilärakenteet aiheuttavat lisääntynyttä painerasitusta jalkapohjiin altistaen jalkapohjavaurioille. Munintakanajalosteilla on myös erilainen alttius eri jalkapohjamuutoksille. Huolimatta jalkapohjamuutosten yleisyydestä ja niiden hyvinvointiongelmasta munantuotannossa, harrastekanoilla jalkapohjamuutosten esiintyvyyttä ei tietääksemme ole aikaisemmin tutkittu.
Tutkimuksen tarkoituksena oli selvittää jalkapohjamuutosten esiintyvyyttä suomalaisilla harrastekanoilla ja jalkapohjamuutosten esiintyvyyteen vaikuttavia tekijöitä. Tutkimus tehtiin suomalaisissa yksityishenkilöiden pitämissä harrastekanaloissa. Jalkapohjamuutoksettutkittin eläviltä linnuilta silmämääräisesti ja arviointiasteikkona sovellettiin viisiportaista broilerien Welfare Quality (WQ)-asteikkoa. Tuloksissa huomioitiin muuan muassa ikä, sukupuoli ja rotu. Kanarodut jaettiin maatiaisiin, puhdasrotuisiin, jalosteisiin ja sekarotuisiin. Maatiaiseksi luokiteltiin suomalaiset alkuperäiskanakannat, puhdasrotuisiksi ulkomaalaiset tuontirodut ja alkuperäisrodut, jalosteiksi tuotannossa käytettävät munintakanajalosteet ja sekarotuisiksi edellä mainittujen rotujen sekoitukset ja kanat, joiden rotu oli tuntematon. Kanalan kuivikkeen kunto arvioitiin kolmesta eri kohdasta: juomakuppien kohdalta, orsien alta sekä vapaalta alalta broilereiden WQ:n viisiportaisella asteikolla.
Tutkimukseen osallistui 14 harrastekanalaa ja yhteensä 193 lintua, joista 140 oli kanoja, 49 kukkoja ja neljän linnun sukupuoli jäi tutkimushetkellä epävarmaksi. Maatiaisia oli 115, puhdasrotuisia 36, jalosteita 10 ja sekarotuisia 32. Nuorin lintu oli 6 viikon ja vanhin 8 vuoden ikäinen.
Yli puolella tutkituista kanoista jalkapohjat arvioitiin terveiksi (jalkapohja-arvioiden keskiarvo 0,7 ± 0,07). Yleisin jalkapohjamuutos oli ihon sarveistuminen ja vakavinta jalkapohjapaisemuutosta esiintyi vähän. Sekarotuisten jalkapohjat olivat terveemmät kuin jalosteilla (p=0,001), puhdasrotuisilla (p=0,003) ja maaaisilla (p=0,013). Kukoilla jalkapohjamuutoksia todettiin vähemmän kuin kanoilla (keskiarvo kukoilla 0,42 ± 0,13 ja kanoilla 0,82 ± 0,08; p=0,009).
Puolella kanaloista oli käytössä pääsääntöisesti kerran vuodessa vaihdettava kestokuivike. Kestokuivike oli usein oljen ja turpeen sekoitus. Muissa kanaloissa kuivikemateriaali oli joko kutterinpuru, olkipelletti tai sahanpuru. Kuivikepohja vaihdettiin hajun, kosteuden ja likaisuuden perusteella useamman kerran vuodessa. Valtaosassa kanaloita kuivike todettiin kuivaksi (kuivikkeiden kunnon keskiarvo 0,74 ± 0,23). Kuivikemateriaali tai kestokuivikkeen käyttö ei vaikuttanut kuivikepohjan kuntoon eri arviointikohdissa. Kuivikemateriaalilla tai kestokuivikkeen käytöllä ei ollut vaikutusta jalkapohjaterveyteen.
Ikä vaikutti jalkapohjaterveyteen, mutta parittainen vertailu ei löytänyt johdonmukaisia eroja ikäryhmien välillä.
Vakavia jalkapohjapaiseita todettiin harrastekanoilla vähemmän kuin tuotantokanatutkimuksissa. Parempi jalkapohjaterveys voisi selittyä tutkimukseen osallistuneiden harrastekanaloiden kuivalla kuivikepohjalla. Selitys kanarotujen välisiin eroihin jalkapohjaterveydessä saattaisi löytyä perimästä ja sen vaikutuksesta jalkapohjien kokoon ja muninnan tehokkuuteen. Jalosteilla todettu huono jalkapohjaterveys voisi osittain johtua tehokkaasta tuotannosta verrattuna vähemmän muniviin rotuihin. Harrastekanaloissa myös munintakanajalosteiden tuotanto jää alhaisemmaksi kuin tuotantokanaloissa johtuen mm. vähemmän tehokkaasta ruokinnasta. Tämä voisi selittää paremman jalkapohjaterveyden kuin samoilla jalosteilla tuotanto-olosuhteissa.
Muninnan puuttuminen voi osaltaan myös selittää kukkojen parempaa jalkapohjaterveyttä. Kukoilla suurempi jalkapohja ja paksumpi anturan rasvapatja saattavat myös vähentää jalkapohjiin kohdistuvaa painerasitusta ja suojata vaurioilta. Kukkojen jalkapohjaterveyttä on tutkittu vain vähän ja yleensä tutkimuksissa on keskityy broileriemoparvien siitoskukkoihin vaihtelevin tuloksin. Kukkojen paremmasta jalkapohjaterveydestä ei tietääksemme ole aikaisemmin raportoitu. Jalkapohjamuutokset lisääntyvät iän myötä ja jalkapohjatulehduksia -ja paiseita voi ilmetä eniten munantuotantohuipun aikaan. Tutkimuksissa on kuitenkin tutkittu vain samanikäisiä, vanhimmillaan 100 viikon ikäisiä munintakanaparvia. Harrastekanaloissa parvien ikähaarukka oli laaja ja siksi tutkimuksen tuloksia ei voi suoraan verrata aiempien tutkimusten tuloksiin.
Tutkimus tarjoaa uutta tietoa harrastekanojen jalkapohjaterveydestä ja osoittaa, että jalkapohjaterveyteen vaikuttavat sukupuoli ja rotu. Harrastekanojen jalkapohjaterveydestä ja jalkapohjamuutosten syntyyn vaikuttavista tekijöistä tarvitaan lisätutkimusta.
Non-steroidal anti-inflammatory drugs (NSAIDs) are the most commonly prescribed analgesics in veterinary medicine. NSAIDs are thought to influence the composition and function of the gut microbiota. As little is known about the recovery time of the gut microbiome post NSAID-treatment, we compared feces microbiome of sows treated with NSAIDs to that of control sows.
Twenty-two sows (parity range 1-5) on one farm were divided into three groups based on their NSAID medication history: never treated (control group, n=9), treated within a year (n=7), and over a year before sampling (n=6). Feces samples were cultured for Lactobacillaceae, which were identified with a colony PCR, and species were identified by comparing 16S PCR product sequences to the National Center for Biotechnology Information’s (NCBI) Basic Local Alignment Search Tool (BLAST) database with 96% accuracy.
Kruskal-Wallis (SPSS) was used to find a difference in Lactobacillaceae species richness (observed) and total count (log10) between the groups.
The Kruskal-Wallis test was found to be significant for culturable Lactobacillaceae species richness (H = 5.92, p = 0.05). The significant difference was seen between the control group and the sows treated within a year before sampling (p = 0.02 and adj. p = 0.05).
The total count of Lactobacillaceae tended to be different between the same two groups before adjusting the p-value (p=0.06).
These results shed light on the role of NSAIDs in the swine gut microbiome. In this study, the reduction of Lactobacillaceae seems to be linked to the use of NSAIDs. During the first year post treatment, richness of culturable species decreased significantly. A tendency for decrease was seen also in the total Lactobacillaceae count. While we acknowledge, that pain relief is vital to improve animal wellbeing, the sows might benefit from probiotic support during and after NSAID treatment to reinforce the recovery of the gut microbiota.
Tail biting is a common, multifactorial behavioural problem in pigs. Although the phenomenon is mostdramatic in fattening pigs, the behaviour often begins in the nursery, and should be prevented already at that stage. This study aimed to establish risk factors for tail lesions in weaners.We performed one-day visits to ten Finnish farms, rearing undocked pigs between approximately 4 weeks to 30 kg. Tail health was observed in in altogether 81 rooms with 1217 pens in terms of 1) lesions (TL, visible lesion, not requiring skin penetration), 2) hanging (HaT) and 3) healed, shortened tails (HST). Each pen was categorized according to the prevalence of each in no, ≤ 10% or > 10% of pigs. The data was expressed on room level as the percentage of pens in each variable. A risk assessment protocol was completed in 35 rooms representing three different age groups.
Tail health data was reduced into three principal components characterizing the different types of tail findings in the sample, together explaining 84.5% of variance in the data. The first component was named “Acute high-prevalence tail biting” (HIGHPREV) as it was loaded on by >10% TL (load 0.75) and >10% Hat (0.45). The component “Underlying tail biting” (UNDERLYING) was loaded on by ≤ 10% (0.68) and >10% HaT (0.71). The component “Acute low-frequency tail biting” (LOWPREV) included ≤10% TL (0.88) and ≤10% HaT (0.46).
Linear mixed models were used to analyse effects of risk factors on room-level tail health measured by component scores. A higher score indicated a higher prevalence of findings. The farm effect was controlled for. HIGHPREV increased with increasing age (p<0.001) and a decreasing lying area temperature (p=0.033).
Effects of enrichment objects were unexpected, as lack of objects was favourable (p=0.014), as was “dirty objects in some pens” as compared to “clean objects in all pens” (p=0.015). The latter may be explained by more use of objects in the “dirty” case. Pens without objects probably had more other types of enrichment.
LOWPREV was unaffected by age or any other risk factor than room size (p=0.02), for which larger was unfavourable. The farm effect was significant (p<0.001), indicating that factors other than those included in the protocol were important.
UNDERLYING was decreased by increasing space allowance (p=0.009) and better resource accessibility for disadvantaged individuals (p=0.02), as well as an interaction between age and availability of feed (p=0.007). The latter was categorized as 1) feed always available (i.e. never emptying feeder, ALW), 2) simultaneous (i.e. long trough with space for all individuals, SIM), and 3) restricted (RES). According to the interaction there was an effect only in the oldest age group (5.-6. nursery week), when ALW was more favourable than both SIM and especially RES. This effect is probably caused by crowding.Resource availability and environmental conditions were the most influential factors predicting tail health in weaners, indicating that it still is relevant to educate farmers on these basic concepts. Farmspecific factors appears, however, also to be important.
Hengitystietulehdusten oireisiin käytetään vasikkakasvattamoissa paljon mikrobilääkkeitä. Virusten ja bakteerien aiheuttamia tulehduksia ei perinteisesti ole pyritty erottamaan, vaan hoito kohdistetaan jälkimmäiseen. Tämä on vastuullisen mikrobilääkekäytön näkökulmasta ongelmallista. Ultraääni soveltuu erittäin hyvin vasikoiden keuhkosairauksien kenttädiagnostiikkaan. Tämän tutkimuksen tarkoituksena oli selvittää yhteyksiä kliinisten oireiden ja keuhkojen ultraäänilöydösten välillä.
174 tapauksen aineisto kerättiin kolmessa vasikkakasvattamossa lokakuun 2022 ja maaliskuun 2023välillä. Otoksessa oli sekä potilaita että tutkimustarkoituksessa valittuja eläimiä. Ilman kliinisiä hengitystietulehduksen oireita tutkittuja kontrollieläimiä oli 31,6%. Otos painoittui 1-2 kuukauden ikäisiin vasikoihin vaihteluvälin ollessa noin yhdestä viikosta kuuteen kuukauteen. Eläimen kliiniset oireet havainnoitiin ennen ultraäänitutkimusta, joka tehtiin Butterfly IQ+ Vet -laitteella keuhkoasetuksin. Keuhkoista tutkittiin kraniaaliosa mahdollisimman laajasti, mediaaliosa kokonaan sekä kaudaalisesti 2-3 kylkiluuväliä. Löydökset kirjatiin heti tai noin puolessa tapauksista myöhemmin videokuvaan perustuen.
Koko aineiston löydöksiin perustuen tehtiin seitsemän ultraäänidiagnoosia (myöhemmin”diagnoosi”). Tiivistymälöydöksen kohdalla tehtiin jaottelu todennäköisen aiheuttajan mukaan suurimpaan läpimittaan perustuen: 1) Bakteeripneumoniassa (44% eläimistä) yli 1,5 cm, 2) bakteeri/viruspneumoniassa (6,6%) 1-1,5 cm ja 3) viruspneumoniassa (8,4%) alle 1 cm. 4) Pleuramuutos-diagnoosi (8,4%) asetettin, jos pleuralinja oli selvästi epätasainen, rikkonainen ja/tai paksuuntunut. 5) Lievä pleuramuutos (19,2%) tarkoitti havaittavaa epätasaisuutta. 6) White lung-diagnoosissa (3,6%) kuvaa hallitsivat ödeemasta kertovat kuvavääristymät (ns. B-lines). Löydösten puutteessa kirjattin 7) Normaali (9,6%). Useita löydöksiä sisältävissä tapauksissa valittiin pienimmän numeron diagnoosi.
Kliinisten oireiden ja diagnoosin yhteyksiä tutkittiin Kruskal-Wallisin testillä kategoristen muuttujien, ja mediaanitestillä rektaalilämmön kohdalla. Parittaiset testit tulkittiin Bonferroni-korjauksen jälkeen.
Rektaalilämpö (n=80), sierainvuodon esiintyvyys (asteikolla kyllä/ ei, n=163) tai korvien asento (asteikolla normaali/ ei, n=157) eivät eronneet tilastollisesti diagnoosiryhmien välillä. Yskäoire oli tavallisempi diagnoosilla Bakteeripneumonia (70,8%:lla diagnoosin eläimistä havaittiin yskä) kuin Lieviä pleuramuutoksia (17,9%, p<0,001, n=117, asteikko kyllä/ei). Silmävuodon ja diagnoosin välillä oli yhteys (p=0,049, n=167, asteikko kyllä/ei), mutta parittaisissa testeissä ei ollut merkitseviä eroja. Silmävuodon esiintyvyys oli matala Bakteeri- sekä Bakteeri/viruspneumoniassa (4,1 ja 0,0%), sen ollessa Normaali-diagnoosissakin 12,5%. Hengitystapa (Kruskal-Wallis p=0,004, n=104) arvioitiin epänormaaliksi useammin Bakteeripneumoniassa (91,1%) verrattuna sekä Lieviin pleuramuutoksiin (50,0%, parittaisen testin p=0,01) eä Normaaliin (40,0%,p=0,02). Korkea hengitystiheys, tässä datassa yli 45 kertaa minuutissa, oli ei-merkitsevästi yhteydessä diagnoosiin (p=0,05, n=151). Korkea hengitystiheys oli tavallisinta Bakteeripneumoniassa (46,3%) ja muuttui odotetulla tavalla diagnoosin vakavuuden mukaan. Hengitysäänten muutokset neliportaisella asteikolla normaaleista (0) voimakkaasti muuttuneisiin (3) olivat yhteydessä diagnoosiin (p<0,001, n=158). Bakteeripneumonia-diagnoosin sisällä painottuivat kategoriat 1-3, kun Lievien pleuramuutosten ja Normaalin ryhmissä painottuivat 0-1 (parittaisten testien p≤0,003).
Saadut tulokset ovat varsin tarkka toisinto ulkomaisista tutkimuksista: oireet antavat hengitystiediagnostiikassa suuntaa, mutta ne ovat vastuullisen antibioottikäytön näkökulmasta epätarkat. Kirjallisuuden suositusten perusteella vain diagnoosi Bakteeripneumonia olisi ollut tutkimushetkellä mikrobilääkehoidon tarpeessa. Bakteeripneumonia olisi tässä datassa löytynyt hyvin muuuneen hengitystavan tai kohtuudella yskäoireen perusteella. Nämä oireet olivat kuitenkin yleisiä muissakin diagnoosityhmissä. Jopa (keuhkoiltaan) Normaaleilla havaittiin yleisesti yskää (25%), sierainvuotoa (25%) muuttunutta hengitystapaa (40%) ja lievästi tai kohtuullisesti muuttuneita hengitysääniä (50,0% ja 7,1%). Ylähengitystietulehdus saattaa ainakin osittainselittää näitä löydöksiä. Tiivistymiä on saattanut jäädä löytymättä, sillä arviolta 5% vasikoiden bronkopneumonioista ei ulotu keuhkojen pinnalle eivätkä siten ole havaittavissa, eikä keuhkoja tutkittu aivan kokonaan.
Tutkimustuloksen perusteella lääkityspäätösten tekeminen oireisiin perustuen saattaa aiheuttaamikrobilääkkeiden liikakäyttöä. Hengitysteiden virusinfektion bakteeripneumoniaksi etenemisen todennäköisyydestä vasikalla tarvitaan enemmän tietoa.
Tutkimusta ovat tukeneet Suomen Eläinlääketieteen Säätiö, Atria Oyj ja Helsingin Yliopisto.
In Finland dairy bull calves are typically transported at an early age from dairy farms either to specialized calf rearing farms or to fatening farms. Transportime is a known risk factor for increased morbidity in calves. Bovine Respiratory Disease (BRD) is one of the main health concerns in the calf rearing units causing increased use of antibiotics and high mortality in pre-weaned calves. It is often associated with transport stress. The aim of the study was to find out how transport time affects the time until the first antimicrobial treatment of the calves that were transported to a rearing unit.The data was collected from transporting bull calves in seven different batches from multiple dairy farms to one rearing unit between January and August 2017. According to the national guidelines, only clinically healthy calves were accepted for transportation. In the vehicles, free access to water and bedding were offered. The rearing unit operated on an all-in all-out basis and the calves were transported to the finishing unit at average age of five months. The transport time from loading a calf at the dairy farm to unloading it at the rearing unit was recorded. The calves were examined by veterinarians one to three days after arrival to the rearing unit and BRD was diagnosed by the BRD scoring system. Antibiotic treatments administered at the rearing unit for each calf were recorded.
Statistical analysis was performed by using STATA/MP 17 for Windows (StataCorp LP, Texas, USA). A p- value < 0.05 was considered statistically significant. The batch was used as a random factor in a model. A linear mixed regression model was used to study the association between transport time and time to the first antimicrobial treatment. Vaccination status, breed, BRD diagnosis at the clinical examination and age and weight of a calf at the arrival were independent variables in the model used. The calves treated with antibiotics at the rearing unit were included in the linear regression model.
Altogether 333 bull calves (174 AY, 159 HOL) were enrolled in the study. At the time of transportation to the rearing unit, they were 10 to 34 days old (mean age 17.0 days, median age 16.0 days). Transport time varied from 45 min to 13 h 25 min (average transport time 6 h 42 min, median transport time 6 h 38 min). 210 (105 AY, 105 HOL) calves were treated with antibiotics at least once during the study period. The time to the first anbioc treatment varied from 0 days to 93 days (average time 21 days, median time 15 days). The longer transport time (hours) decreased the time to the first antibiotic treatment (days) (coef -0.942, p=0.044.). Other variables studied were not significant. Though the transportation distances in Finland are moderate, the longer transport time decreased the time until the first antimicrobial treatment. The longer transport time might have effect on subclinical respiratory infecons becoming clinical diseases.
Proliferative enteropathy is caused by Lawsonia intracellularis (LI), which is a common enteric pathogen found in pig production worldwide and, according to Finnish Food Authority*, also in Finnish pigs. However, so far, a LI prevalence study has not been conducted in the Finnish conventional pig herds.
In 2022, two studies were conducted with the objective to investigate the prevalence (study 1) and level of LI and Brachyspira pilosicoli (BP) as well as the LI seroprevalence in Finnish grower and slaughter pigs (study 2), respectively.
In study 1, one faecal sock sample was collected from batches of pigs weighing 25-35 kg in 36 farms with a history of diarrhea outbreaks. No antimicrobial treatments were given at time of sampling. qPCR was performed (at L&F, Veterinary Laboratory, Denmark) to determine the bacterial shedding of LI and BP, having both qPCR tests with lower detection limit of 3 and 2 log(10) copies/gram feces, respectively.
In study 2, blood samples were collected at three major Finnish slaughterhouses by random sampling during one-two working days (100 samples from 10 farms in 1 slaughterhouse; 150-155 samples from 15 farms in each of 2 slaughterhouses). Samples were analysed for LI antibodies by ELISA (Svanova®) at CDS, The Netherlands; cut-off value for positive samples at ≥30% inhibition was used.
Prevalence and mean excretion levels in positive fecal samples (median; range) were 61.1% and 5.13 (5.29; 3.04-6.05) for LI and 44.4% and 4.04 (4.17; 2.00-6.40) for BP, both log(10) copies/gram of feces. On average 217 pigs were sampled in each batch (median: 180; range: 40-588). In total, 405 samples from 45 farms were collected for LI seroprevalence. Farm seroprevalence was 100%, whereas pig prevalence was 93.6% (variation between slaughterhouses: 91.3-96.8%). Using faecal sock samples, both LI and BP were commonly found in Finnish grower pigs, though LI was more common than BP. Mean excretion of LI were found at levels known to cause proliferative lesions in the intestine and to potentially cause a loss in productivity. All finisher farms were seropositive for LI with a high prevalence, suggesting that infection becomes apparent during the grower-finisher period.
*Finnish Food Authority; Animal Diseases in Finland 2021. August 2021. foodauthority.fi
Campylobacter species are commonly found in the intestines of asymptomatic sheep, but in some cases, it can cause symptoms, including spontaneous abortions. There is no comprehensive information on the prevalence of Campylobacter in sheep in Finland. However, studies from other countries suggest sheep as an important source of human Campylobacter infections. The aim of this study was to comprehensively investigate the prevalence and genotype distribution of Campylobacter in sheep farms and whether the prevalence of Campylobacter in sheep differs between animals of different age groups, mainland versus waterside pastures, or different times of the year. A total of 231 composite fecal samples were studied from 19 sheep farms—ten farms with waterside pastures and nine farms with pastures without direct access to surface water. Samples were taken on four separate occasions on each farm: February, May, August, and November in 2023. Furthermore, the sheep were divided into five sampling groups: ewes, rams, lambs (less than 4 months old), ewe lambs (4-12 months old), and ram lambs (4-12 months old). Campylobacter spp. were isolated using both a direct culture on mCCDA at 37°C and enrichment using Preston broth followed by culture onto mCCDA selective agar at 41.5°C. Preliminary genus-specific identification was performed using real-time PCR. DNA isolated from pure Campylobacter spp. cultures (n=43) from 30 samples were sent for whole-genome sequencing. Genotyping for Campylobacter jejuni and Campylobacter coli was conducted using multilocus sequence typing (MLST), and other Campylobacter species were identified using Microbial Genomes Atlas Online (MiGA) and a core genome phylogeny against type strains from all validly published and recently described sp. nov. of Campylobacter. The overall prevalence of Campylobacter spp. in sheep was 19%, being lowest in spring (10% in February, 1.7% in May) and highest in autumn (36% in August, 28% in November). C. jejuni was the most common species representing 58% (n=25) of the sequenced isolates, followed by Campylobacter ovis 26% (n=11), C. coli 7% (n=3), other Campylobacter sp. 4.7% (n=2), Campylobacter lari 2.3% (n=1) and Campylobacter upsaliensis 2.3% (n=1). We identified 8 different sequence types (ST) among the C. jejuni isolates, including one novel ST. The most common ST was ST-1701, comprising 36% of the C. jejuni isolates from three farms. All C. coli isolates were identified as ST-827, while C. lari was ST-21. Animals of different age groups from individual sheep farms mostly exhibited different STs and different Campylobacter species at each sampling, indicating a diverse and dynamic Campylobacter population at the sheep farms. In conclusion, a clear seasonal variation was observed in the prevalence of Campylobacter in sheep feces, with lower prevalence during the indoor feeding period in spring and significantly higher prevalence in autumn suggesting Campylobacter carriage is associated with grazing and increased exposure to the environment. The funding for the study came from the Lammaslaidun-project funded by Makera (Agricultural Development Fund by The Ministry of Agriculture and Forestry).
Suomessa on tehty tautisuojauksen arviointikäyntejä sikatiloilla vuodesta 2019 lähtien (Biocheck.Ugent®- arviointikäynti, joka kehitetty Gentin Yliopistossa). Vuosittaisen Biocheck-arviointikäynnin puuttuminen on vaikuttanut pitopaikan terveysluokitukseen Sikavassa vuodesta 2021 alkaen. Sikavassa oli vuoden 2023 lopussa 711 aktiivista pitopaikkaa, joista 669:lle oli tehty vähintään yksi Biocheck -arviointikäynti. Sikavan pitopaikkojen kokonaistulosten keskiarvo on parantunut joka vuosi, mutta joissain tautisuojauksen perusasioissa olisi edelleen parantamisen varaa.
Sellaisten pitopaikkojen määrä, joissa on tautisulku, on lisääntynyt joka vuosi. Yli kolmasosalla pitopaikoista (38 %) ei kuitenkaan ole sellaista tautisulkua, joka erottelisi selkeästi puhtaan ja likaisen puolen. Toimimaton tautisulku voi mahdollistaa taudinaiheuttajien pääsyn sikalaan. Yllättävän monella pitopaikalla (37 %) kävijöiden ei tarvitse pestä ja/tai desinfioida käsiä ennen eläintilaan menoa ja tältä osin tilanne suomalaisilla pitopaikoilla ei ole juuri parantunut. Tämä lisää tarttuvien tautien, kuten influenssan ja jopa salmonellan leviämisen riskiä. Tilakeskuksen rakennuksien seinustojen siisteys on parantunut vuosi vuodelta, mutta edelleen 36 %:lla pitopaikoista seinustoilla on kasvillisuutta tai säilytetään tavaraa. Tämä lisää haittaeläinten esiintyvyyden riskiä ja juuri haittaeläimien epäillään olevan lähes kaikkien viime vuosien salmonellatapauksien taustalla. Edelleen 34 %:lla pitopaikoista ei ole käytössä lastaustilaa ja viime vuosina kehitystä tämän suhteen ei ole tapahtunut. Yhdellä kerralla tyhjennettävässä sikalassa lastaustila ei ole niin tärkeä. Vuoroimetystä tai apuemakkoa käytetään, kun porsaita syntyy enemmän verrattuna emakon imetyskykyyn. Jos pahnueita tasataan, pitää huolehtia siitä, että porsaita siirretään vain tarvittaessa ja yhtä porsasta ei siirretä kuin kerran. Siirrot ovat pikkuporsaille hyvin kuormittavia ja altistavat sairauksille. Biocheck-käyntien tuloksien mukaan jopa 12 %:lla pitopaikoista siirretään porsaita useammin kuin kerran. Huolestuttavasti näyttää myös siltä, että siirtelyt ovat viime vuosina olleet lisääntymään päin.
Vain 41 %:lla pitopaikoista vieroitettujen porsaiden osastot olivat kertatäyttöisiä (mukana pitopaikat, joilla vieroitetuille on erilliset osastot). Lihasikojen osastot olivat kertatäyttöisiä 57 %:lla sellaisista pitopaikoista, joilla on lihasikoja. Jatkuvatäyttöisissä osastoissa erätaukopesua ja -desinfiointia ei pystytä tekemään. 19 % pitopaikoista on ilmoittanut vuonna 2023, että pitopaikan läheisyydessä (10 km) on havaittu villisikoja. Näistä tiloista vain 5 % (6 kpl) on aidannut pitopaikan tuotantorakennukset. Jos afrikkalainen sikarutto (ASF) leviäisi Suomen villisikoihin, voisi tauti levitä niistä tuotantosikaloihin. Tämä riski kasvaa, jos villisiat pääsevät tilan läheisyyteen ja henkilöliikenteen tautisuojauksessa tai rehujen ja kuivikkeiden säilytyksessä on puutteita tautisuojaukseen liittyen. Sikatiloilla on hyvä olla suunnitelma aitaamiselle, sillä jos ASF todettaisiin Suomessa villisioissa, komission määräämällä tartuntavyöhykkeellä tuotantorakennusten tulee olla aidattuja.
Sikavassa olevien pitopaikkojen tautisuojauksen taso on hienosti noussut vuosittain, mutta monessa pitopaikassa on vielä parannettavaakin. Säännöllisten Biocheck-arviointikäyntien avulla löydämme pitopaikkojen kehityskohteita ja voimme varmistua, että kehitystä tapahtuu. Näin saamme kehitettyä yksittäisten pitopaikkojen sisäisen ja ulkoisen tautisuojauksen tasoa ja pidämme yllä Suomen hyvää tautitilannetta.
Utaretulehdus on laajasti levinnyt ja taloudellisesti merkittävä lehmien hyvinvointia ja maidontuotantoa vähentävä tartuntatauti. Suurin osa lypsylehmillä käytetyistä mikrobilääkehoidoista käytetään utaretulehduksen hoitoon. Kansainvälisesti mikrobilääkehoitojen käyttöä pyritään vähentämään lääkeresistenssin hillitsemiseksi. Utaretulehduksen ennaltaehkäisyssä lypsyn jälkeinen vedinten käsittely vedinkastoaineella on merkittävä toimenpide vedinten iholla elävien utaretulehdusmikrobien eliminoinniksi. Nykyiset vedinkaston tehoaineet voivat kuitenkin olla silmiä tai ihoa ärsyttäviä, jodipitoiset yhdisteet ovat myrkyllisiä vesieliöille ja klooripohjaiset yhdisteet voivat muodostaa myrkyllisiä kaasuja. Variksenmarjan solukon sisällä elävistä bakteereista eristetyt antimikrobiset peptidit (myöh. peptidi) voisivat toimia tehoaineena vedinkastoliuoksessa. Niillä voi olla myös käyttökohteita mikrobilääkkeiden korvaajana kotieläintuotannossa. Tutkimuksessa selvitettiin peptidien utaretulehdusmikrobien kasvunestävää tehoa vedinkastoliuoksessa laboratorio-olosuhteissa.
Viiden peptidin bakteriostaattista vaikutusta tutkittiin utaretulehdusta aiheuttaville mikrobeille Staphylococcus aureus, haemolyticus, simulans, epidermis ja chromogenes, Streptococcus uberis, Streptococcus dysgalactiae, Candida albicans ja Cryptococcus neoformans. Mikrobien liuoskasvatukset tehtiin kuoppalevyillä pääosin Bioscreen C° Pro (Growth Curves Ltd., Suomi) -laitteessa, joka kasvatuksen aikana mittasi kasvatusliuosten sameuden joka 15 minuutti. Kaikki mikrobikasvatukset tehtiin 37 °C:ssa ja mikrobien alkukonsentraationa käytettiin 500 000 bakteerisolua/ml. Peptidejä ja verrokkina gentamisiiniliuosta lisättiin eri pitoisuuksissa (0–128 μg/ml) kuoppalevylle kasvatusliuoksiin. Kasvuston mitattua sameutta käytettiin mikrobikonsentraation ja pienimmän mikrobin kasvua kokonaan 24 tunniksi estävän peptidipitoisuuden (MIC-arvo) määrittämiseen. Peptidien yhteisvaikutusta klooriheksidiinin kanssa tutkittiin lisäämällä kuoppalevylle eri pitoisuuksia peptidejä (0–16 μg/ml) ja klooriheksidiiniliuosta (0–2 μg/ml) Staphylococcus aureus kasvatusliuokseen. Peptidien ja klooriheksidiinin MIC-arvo kasvustoissa määritettiin 24 ja 48 tunnin kasvatuksen jälkeen sekä näytteissä, joihin oli lisätty vain yhtä tutkittavista yhdisteistä, että katsoen minkälaisien pitoisuuksien yhdistelminä yhdisteet estivät mikrobin kasvun. MIC-arvojen avulla laskettiin FIC-arvo (Fractional inhibitory concentration) ja FIC-indeksi (FICI), jotka kertovat vaikuttivatko tutkitut yhdisteet yhdessä enemmän kuin erikseen. Yhteisvaikutusta Staphylococcus aureus kasvuun tutkittiin tarkentavasti kuoppadiffuusiokokeessa. Bakteerilaimennoksen pipetoinnin jälkeen elatusainemaljalle tehtiin teräväkärkisellä, steriilillä agarporalla kuopat, joihin pipetoitiin klooriheksidiiniliuosta (0–2 μg/ml) eri peptidipitoisuuksissa (0–200 μg/ml) ja verrokkina gentamisiiniliuosta (5 μg/ml). Bakteerin kasvua havainnoitiin ja estorenkaan (mm) säde mitattiin 24 tunnin inkubaatioajan jälkeen. Peptidien säilyvyysaikoja eri vedinkastokoostumuksissa tutkittiin säilyttämällä liuoksia huoneenlämmössä ja jääkaappilämpötilassa (4 °C) viikon sekä yhden, kahden, kolmen ja kuuden kuukauden ajan. Säilytysaikojen jälkeen peptidi-vedinkastoliuokset lisättiin Staphylococcus aureus kasvatusliuokseen ja seurattiin jälleen viljelmien sameutta. Verrokkina kokeessa käytettiin vedinkastoliuoksia ilman peptidejä sekä puhdasta peptidien puskuriliuosta.
Tutkituista peptideistä kaksi olivat tehokkaimpia ja toimivat laaja-alaisimmin testattuja mikrobeja vastaan. Niiden MIC-arvot testatuille mikrobeille olivat 1–8 μg/ml vastaten WHO:n tärkeimpien antimikrobilääkkeiden listalla olevan gentamisiinin MIC-arvoa. Peptidien ja klooriheksidiinin FICI oli ≤0,5 osoittaen Staphylococcus aureus bakteerin kasvua estävän synergisen vaikutuksen kuoppalevykokeessa. Synerginen vaikutus todettiin myös kuoppadiffuusiokokeessa estorenkaiden ollessa 2–4 mm. Peptidit säilyttivät tehokkaasti mikrobien kasvunestävän vaikutuksensa erilaisissa vedinkastokoostumuksissa ja säilytyslämpötiloissa kuuden säilytyskuukauden jälkeenkin. Parhaiten teho säilyi peptidien ja klooriheksidiini-pohjaisen vedinkaston yhteisliuoksessa.
Antimikrobiset peptidit voivat toimia vedinkastoliuoksen utaretulehdusmikrobien kasvunestävänä yhdisteenä. Jatkotutkimuksia tarvitaan antimikrobisten peptidien muiden käyttökohteiden, jotka mahdollistavat haitallisten desinfiointiyhdisteiden ja mikrobilääkkeiden käytön vähentämisen kotieläintuotannossa, kehittämiseksi sekä peptidien tehon osoittamiseksi kenttäkokeissa. Peptidien hyödyntämiseksi kaupallisissa tuotteissa niiden tulee myös läpäistä EU:n valvontaviranomaisten hyväksymismenettelyt, jotka vaihtelevat käyttötarkoituksen mukaan.
The aim was to investigate the effect of an alpha2-adrenoceptor antagonist (vatinoxan) on serum insulin and blood glucose (BG) concentrations in horses after intravenous glucose administration. Nine Finnhorses were assigned to two intravenous treatments in cross-over design: 1. saline (SAL) and 2. vatinoxan HCl (0.2 mg/kg; VAT). The horses were fasted overnight. Immediately after administration of an intravenous glucose bolus (150 mg/kg), the treatment was given as a single IV bolus via jugular catheter. Venous blood samples were taken for insulin and BG before glucose and until 300 minutes after treatment. Blood glucose was analyzed immediately with a glucometer (Alpha-TRAK II). Serum was frozen until analyzed for insulin by an ELISA kit. The differences between treatments were evaluated with RM ANCOVA.
Administration of IV glucose (SAL) resulted in a rapid increase in serum insulin and BG. The peak concentrations (mean insulin 30.9±9.3 μIU/mL; mean glucose 12.3±0.9 mmol/L) were detected at 15 min after treatment. After VAT, peak concentrations (mean insulin 40.9±15.3 μIU/mL; mean glucose 11.0±1.2 mmol/L) were detected at 15 min.
The increase from baseline in insulin was significantly larger at 15 min after VAT (p = 0.04) than after SAL. The increase from baseline in BG was significantly less (p=0.005) after VAT than after SAL. Vatinoxan alleviated the increase in BG after IV glucose administration. Therefore, alpha2-adrenoceptors may play a role in regulation of equine carbohydrate metabolism, but more research is warranted.
This study was approved by the National Animal Experimentation Board of Finland. The authors would like to thank Finnish Foundation of Veterinary Research and Erkki Rajakoski foundation (Hippos ry) for funding this study and Vetcare Ltd. for providing vatinoxan and funding for the experiments.
Parapoxviruses (PPV) cause skin and mucous membrane lesions in several animal species, and are often zoonotic. Equine PPV (EqPPV) is initially described in humans in the United States and later in a severely sick horse in Finland in 2013–2015. In 2021–2022, a large-scale pustulo-vesicular pastern dermatitis outbreak occurred in horses around Finland. This study aimed to identify and analyze the causa:ve agent, describe the disease, and prepare for future epidemics by studying disease transmission and developing diagnostics.
Samples to identify causative agent were taken from skin lesions of acutely ill horses during the outbreak. Epidemiological survey was conducted to gain understanding on the epidemiology, clinical signs, treatment, distribution, risk factors, and zoonotic potential.
Skin samples from 26 horses representing 11 stables were subjected to bacteriological, histological and virological analysis. Epidemiological survey was conducted online and horse farms in Finland were invited to partocipate.
EqPPV was identified as a probable cause of the epidemic and co-infections with several potentiallypathogenic and zoonotic bacteria were observed. Almost complete virus genome with little sequencevariation was sequenced and annotated from several samples. Histopathologically, suppurative and ulcerative dermatitis was diagnosed. Cases were reported in entire Finland. Having racehorses and contact with racecourse were significant risk factors for the infection. Skin symptoms consistent with PPV infection werereported in humans in case stables significantly more open than in control stables.
Samples from the humans taking care of sick horses and presenting typical skin signs for PPV infection were not obtained. Therefore, zoonotic potential of EqPPV could not be confirmed.
The disease spread quickly among racehorses in the entire country in winter 2021-2022. EqPPV had previously only been described in one horse in 2013 and had not been shown to cause epidemics, making it mostly unknown to racing community and clinicians. Reports of skin lesions in humans and zoonotic nature of other PPVs makes this disease a potential zoonosis, highlighting a need for OneHealth approach. The epidemic pinpointed a lack of preparedness to epidemics caused by new pathogens and this study, together with efficient communica:on, enables a beTer preparedness for future outbreaks.
Alpha-2 adrenoceptor agonists are widely used for equine sedation regardless of their side effects. The aim of the study was to investigate the effects of detomidine infusion with and without a peripherally acting alpha-2 adrenoceptor antagonist, vatinoxan, on blood glucose (BG) and insulin concentrations.
Eight Finnhorses were assigned to two 4-hour infusions: detomidine (0.01 mg/kg + 0.015 mg/kg/h IV) (DET) and a combination of DET and vatinoxan (0.15 mg/kg + 0.05 mg/kg/h IV) (DET+VAT) using cross-over design. Blood samples were taken before, during and 4 hours after the infusion at one hour interval. Blood glucose was analyzed immediately after collection with a glucometer (AlphaTRAK2). Serum insulin concentration was analyzed with equine-optimized ELISA (Mercodia equine insulin ELISA).
Mean BG peaked (17.0±2.2 mmol/L) at the end of DET infusion and was higher than with DET+VAT (10.0±1.6 mmol/L, p<0.001). During DET infusion, median insulin concentration was lower than limit of quantification (1.15 μIU/mL, min-max <1.15-1.49 μIU/mL) and lower than with DET+VAT (2.24 μIU/mL, min-max <1.15-4.58 μIU/mL, p=0.018). Insulin concentration did not reach the baseline within 4 hours after the infusion in either group.
In this study, vatinoxan alleviated the detomidine-induced increase in BG and decrease in serum insulin concentration during a 4-hour infusion and may therefore be a beneficial addition to equine sedation protocols. In humans, long-term glycemic variability has been associated with adjunctive diseases in diabetic patients. Additionally acute glucose fluctuations have been shown to cause oxidative stress. However, it is unknown whether a long-lasting hyperglycemia and hypoinsulinemia are detrimental for horses and therefore further studies are warranted.
Equine asthma (EA) is a common noninfectious inflammatory airway disease associated with airwayneutrophilia andairway remodeling. Matrix metalloproteinase 9 (MMP 9) is an enzyme which is predominantly derived fromneutrophils and considered a potential promotor of airway remodeling. Previously, increased activity of MMP 9 has been found in bronchoalveolar lavage fluid (BALF) of asthmatic horses with airway neutrophilia.The aim was to study the expression of MMP9 in endobronchial biopsies (EBB) of horses with naturally occurringasthma of variable degree.
EBB and BALF were obtained from 20 horses with asthma and 19 control horses. BALF wasanalyzed for cytology, whereas EBB were graded for remodeling and stained immunohistochemically for MMP 9.The BALF neutrophil percentage (SD) in horses with asthma was 11.3 (15.9) and in controls 4.4 (5.9), andthe average [range] EBB remodeling score was 8 [5–11] and 6 [2–9], respectively. EBB samples were positive for MMP 9with epithelial intracytoplasmic labelingin both groups of horses.
In this group of horses with naturally occurring EA, the bronchial MMP9 protein expression visualized with immunohistochemistry did not appear to be associated with the extent of airway neutrophilia or airway remodeling.The results will contribute to the study on the role of bronchial epithelium in the pathophysiology of EA.
The purpose of the study was to describe iatrogenic histopathological abnormalities in two equine globes enucleated shortly after therapy with subconjunctival amphotericin B injections.
Clinical records of horses treated for presumed or diagnosed keratomycosis with subconjunctival amphotericin B injections followed by histopathology of the treated globe in the University of Helsinki Veterinary Teaching Hospital between 2017 and 2023 were retrospectively reviewed.
A 9-year-old warmblood mare and a 9-year-old warmblood gelding with clinical findings consistent with a deep corneal stromal abscess were included. In both cases, 0.2 ml of 5 mg/ml amphotericin B (Fungizone, Cheplapharm Arzneimittel GmbH, Germany) reconstitute was administered in the subconjunctival space every other day as a part of the therapeutic protocol with broad-spectrum combination of antimicrobial and antifungal agents for the suspected corneal fungal infection. In both cases, the response to therapy was deemed insufficient and surgical intervention with the aim to salvage the globe was discussed. Both owners declined further treatment. The mare underwent enucleation 4 days after the third injection. The gelding was euthanized according to owner’s decision 8 hours after the second injection.
A deep stromal abscess caused by hyphated fungi was confirmed with histopathology in the affected globes in both cases. Both globes showed a focally extensive subconjunctival and episcleral area of hemorrhagic necrosis with fibrin exudation and cellular debris derived from karyorrhectic neutrophilic granulocytes in the area of the previous injections. In the periphery of the lesion, fibroblast proliferation, neovascularisation, fibrinoid vasculitis and occasional organizing arterial thrombi were appreciated. Special staining for bacteria (Gram) and fungal organisms (GMS; PAS) did not reveal infectious agents in the area affected by necrosis following injections.
In conclusion, subconjunctival 5 mg/ml amphotericin B injections may cause focally extensive sterile necrosis at the injection site. This potential adverse effect should be taken into consideration in clinical decision-making process. Further studies regarding safety and efficacy of this therapy are required.
Adequate image quality in equine echocardiography requires stillness from the patient. Sedation with alpha₂-adrenoceptor agonists may alter cardiac measurements. An alpha₂‐adrenoceptor antagonist, vatinoxan does not cross the blood-brain-barrier. Therefore, it alleviates the peripheral effects of detomidine, while the central sedative effects remain unaffected. This makes vatinoxan a potential addition to sedative protocols for echocardiography in restless horses.
Standard transthoracic M-mode echocardiography of the left ventricle was performed to evaluate internal diameter in systole and diastole (LVIDs, LVIDd) and fractional shortening (FS) at the level of papillary muscles in 6 horses (median age 13 years; range 5-21) without clinical evidence of cardiac disease. Horses with an audible murmur ≥ 2/6, elevated cardiac troponin I or evidence of general illness were excluded. The echocardiographic examination was performed pre-sedation and 10 minutes post-sedation with detomidine HCl 10 μg/kg and vatinoxan HCl 200 μg/kg IV. The effects of sedation were evaluated with T-test or Wilcoxon signed rank test.
Median heart rate was 40 (range 36-44) pre-sedation and 38 (range 36-42) 15 minutes post-sedation. Mean ± SD LVIDs, LVIDd and FS were 5.32 cm ± 0.48, 9.84 cm ± 0.72, 45.92% ± 2.91 pre-sedation and 5.27cm ± 0.66, 9.54 cm ± 0.69, 44.62% ± 6.52 post-sedation, respectively. No significant differences were noticed between any pre- and post-sedation values.
In conclusion, pre- and post-sedation HR, LVID and FS values were comparable in horses sedated with detomidine and vatinoxan. Echocardiographic measurements might be reliably performed under detomidine-vatinoxan sedation in horses.
Background: Sedation in horses is typically achieved using alpha-2 adrenoceptor agonists, although their use is associated with multiple side effects. A peripheral alpha-2 adrenoceptor antagonist, vatinoxan has been shown to alleviate some of them.
Objectives: To evaluate the effects of vatinoxan infusion on gastrointestinal motility in horses sedated with detomidine infusion.
Study Design: Experimental blinded cross-over study.
Methods: Eight horses received two 4-hour infusions: detomidine (0.01 mg/kg + 0.015 mg/kg/h IV) (DET) and a combination of detomidine and vatinoxan (0.15 mg/kg + 0.05 mg/kg/h IV) (DET+VAT). Horses were tubed with plastic marker balls before the start of the infusion. Borborygmi score was monitored during and after the infusion. Excretion of balls, faecal moisture and faecal output were repeatedly monitored for 72 hours after the infusion.
Results: Borborygmi score remained significantly higher during DET+VAT infusion and the following hour compared to DET (p<0.05). Median cumulative weight of faeces during the first 8 hours after the end of infusion was significantly higher for DET+VAT [6.25kg (3.52-8.65)] than DET [2.85kg (1.7-6.6)] (p=0.007). The balls were excreted significantly faster after DET than after DET+VAT infusion (p=0.019). Faecal moisture ranged from 73.5 to 86.0% and did not differ between treatments.
Conclusions: Combining vatinoxan with detomidine significantly improved borborygmi score. Horses treated with DET+VAT passed more faeces in the initial period after the infusion although the excretion of balls was faster with DET. Therefore, vatinoxan may benefit gastrointestinal motility in horses treated with detomidine infusion, although the results were somewhat conflicting.
This research project was kindly sponsored by Vetcare Oy Finland.
Hevosten tulehduksellinen suolistosairaus (inflammatory bowel disease, IBD) on melko tutkittu sairaus, jota on vaikea diagnosoida ja hoitaa. Hoitovasteeseen ja ennusteeseen vaikuttavat tekijät ovat huonosti tunnettuja, vaikka IBD-epäilyjen määrä on lisääntynyt viime vuosien aikana. Tämän tutkimuksen tarkoituksena oli kuvata IBD:n varalta tutkittujen suomalaisten hevosten yleispiirteet, kliiniset löydökset, diagnostiset tutkimukset, peräsuolibiopsian löydökset sekä hoitovaste.
Tutkimukseen valittiin vuonna 2022 IBD:n varalta tutkitut hevoset, joista oli saatavilla hyvänlaatuinen peräsuolibiopsia. Hevosten yleistiedot, oireet, kliiniset ja histologiset löydökset sekä hoitovaste kerättiin retrospektiivisesti omistajilta sekä osallistuvilta klinikoilta. Muuttujien vaikutusta oireisiin, hoitovasteeseen ja biopsiatulokseen arvioitiin tilastollisin menetelmin (khiin neliö -testi, merkitsevyystaso p-arvo < 0,05).
Tutkimukseen valikoitui 152 hevosta. Yleisimmät kliiniset oireet olivat alentunut suorituskyky (68 %), epämääräinen kipu (43 %) sekä ärtyneisyys/aggressiivisuus (41 %). Erilaisia suolisto-oireita todettiin yhteensä 65 % hevosista, joista yleisin oli ripuli (30 % hevosista). Hevosten oireilun suhteen käytösmuutos oli tilastollisesti merkitsevästi yhteydessä siihen, että peräsuolibiopsiassa todettiin muutoksia (p = 0,002). Ultraäänitutkimuksessa muutoksia ohutsuolessa todettiin 66 % hevosista, ja mahahaava todettiin 37 % hevosista. Vain 12 % hevosista oli tehty ontumatutkimus IBD diagnostiikan yhteydessä. Kesäkaudella 81 % hevosista laidunsi osa- tai kokoaikaisesti, ja laiduntavilla hevosilla oireilu merkitsevästi väheni laidunkauden aikana (p = 0,002). Laiduntaminen ei kuitenkaan vaikuttanut hevosen hoitovasteeseen (p = 0,214), eikä peräsuolibiopsian löydös eronnut merkitsevästi niillä hevosilla, jotka oireilivat myös laidunkauden aikana (p = 0,422). Peräsuolibiopsioista 35 % todettiin eosinofiilinen, 18 % lymfoplasmasytäärinen ja 6 % neutrofiilinen tulehdus, lisäksi 40 % ei todettu tulehdusmuutoksia. Tulehdusmuutoksista 78 % oli lieviä ja 20 % kohtalaisia. Tulehdusmuutoksia nähtiin yleisimmin 5–8-vuotiailla hevosilla, kun taas 9–12-vuotiailla hevosilla todettiin vähiten muutoksia peräsuolibiopsiassa.
Yleisimmin IBD-epäillyn hevosen hoitona oli käytetty ruokintamuutosta yhdistettynä lääkehoitoon (68 %). Glukokortikoidit olivat ensivaiheen lääke 73 % hevosista ja lääkitystä muutettiin 28 % hevosista. Hoitovaste arvoitiin hyväksi lähes puolella potilaista (49 %). Kliiniset oireet ja löydökset, biopsialöydös sekä hoitotyyppi eivät olleet yhteydessä hoitovasteeseen. Hevosilla, joilla lääkitystä muutettiin, hoitovaste oli merkitsevästi huonompi (p = 0,011).
IBD-diagnostiikkaa, mukaan lukien peräsuolibiopsian ottoa, tehdään Suomessa usein hevosille, joilla on alentunut suorituskyky, epämääräistä kipuilua tai ärtyneisyyttä/aggressiivisuutta ja joissa ontumatutkimusta tai gastroskopiaa ei ole suoritettu. Nämä oireet ovat epäspesifisiä ja sopivat lukuisaan muuhunkin sairauteen IBD:n lisäksi. Tässä tutkimuksessa vain harva tekijä vaikutti IBD-epäilyssä hoitovasteeseen hevosilla, joten hoitovasteen ennustettavuutta on vaikea parantaa tällä hetkellä käytössä olevilla tutkimuksilla ja hoitovaihtoehdoilla. Kuitenkin usealla tutkimusaineiston hevosella oli vain yksittäinen suolisto-oire eikä hoitovasteessa tai peräsuolibiopsian muutoksissa nähty merkitseviä eroja niissä hevosissa, joilla oli suolisto-oireita verrattaessa hevosiin, joilla ei todettu selviä suolisto-oireita. Näiden perusteella IBD-diagnostiikka voi olla tarpeen myös tapauksissa, jossa suolisto-oireet ovat lieviä tai vähäisiä. Tässä tutkimuksessa peräsuolibiopsian löydöksellä ei ollut yhteyttä hoitovasteeseen, joten peräsuolibiopsian tulos ei yksin ratkaise hoidon tarvetta.
Pulssioksimetriaa veren happisaturaation mittaamisessa on tutkittu hereillä olevilla varsoilla vain vähän eikä menetelmä ole niillä rutiinikäytössä. Pulssioksimetrialla olisi hapetuksen arvioinnissa monia etuja valtimoverinäytteeseen verrattuna; menetelmä olisi kivuton ja ei-invasiivinen, ja happisaturaatiota voitaisiin seurata reaaliaikaisesti. Tämän prospektiivisen kliinisen tutkimuksen tarkoituksena oli arvioida pulssioksimetrian käytettävyyttä, validiteettia ja luotettavuutta happisaturaation mittaamisessa hereillä olevilla varsoilla kahdesta mittauskohdasta (huuli ja takavatsan ihopoimu).
Tutkimuksessa käytettiin kolmeakymmentäkahta alle kuukauden ikäistä varsaa Helsingin yliopiston Yliopistollisessa hevossairaalassa. Yhdeksällätoista varsoista ei ollut hengitys- ja verenkiertoelimistön ongelmia, ja kolmellatoista oli keuhkokuume. Pulssioksimetrimittaukset tehtiin kolmena toistona varsan huulesta ja/tai ihopoimusta (mittausten kokonaismäärä huulesta n=196 ja ihopoimusta n=338) päivittäin varsojen sairaalassaoloaikana. Lisäksi kuudellatoista varsalla mittaukset tehtiin samassa mittaussessiossa sekä huulesta että ihopoimusta mittauskohtien keskinäistä vertailua varten (n=58 mittausta). Varsoilta otettiin myös valtimoverinäytteitä pulssioksimetrimittausten yhteydessä takajalan metatarsaalivaltimosta (n=97). Huulesta ja ihopoimusta samassa mittaussessiossa mitattuja happisaturaatioarvoja verrattiin keskenään, ja happisaturaatioarvoja molemmista mittauskohdista verrattiin myös valtimoverinäytteen perusteella laskettuun happisaturaatioarvoon. Lisäksi pulssioksimetrimittausten toistettavuutta arvioitiin useiden mittaajien (n=5) osalta.
Pulssioksimetrillä mitattu veren happisaturaatio oli keskimäärin (± keskihajonta) 1,8 (±3,3) prosenttiyksikköä korkeampi huulesta ja 5,7 (±4,3) prosenttiyksikköä korkeampi ihopoimusta kuin laskennallinen saturaatio. Kun saman mittaussession saturaatiomittaustuloksia huulesta ja ihopoimusta vertailtiin varsakohtaisesti, ihopoimun mittaukset olivat 2,4 (±2,4) prosenttiyksikköä korkeampia kuin huulen mittaukset.
Pulssioksimetrimittausten toistettavuus oli kohtalainen tai hyvä ja parani merkittävästi, kun kauimpana keskimmäisestä mittausarvosta ollut mittaustulos poistettiin. Useimmiten ensimmäinen mittaustulos oli poikkeavin. Varsat sietivät pulssioksimetrimittaukset hyvin, mutta hereillä olevilla varsoilla liikkeestä johtuvat kosketusongelmat aiheuttivat satunnaisia teknisiä haasteita. Hereillä olevilla varsoilla pulssioksimetria huulesta (mutta ei ihopoimusta) on käyttökelpoinen ja validi menetelmä happisaturaation määrittämiseen kliinisessä työssä. Toistomittaukset lisäävät menetelmän luotettavuutta.
Keuhkokuume on merkittävä sairastuvuuden ja kuolleisuuden syy vastasyntyneillä varsoilla. Näiden varsojen selviytymiseen liittyvä tekijöitä sairaalaolosuhteissa ei ole kuitenkaan vielä juurikaan selvitetty. Retrospektiivisen kliinisen tutkimuksemme tarkoituksena oli tutkia mahdollisia ennustetekijöitä, jotka liittyvät keuhkokuumetta sairastavien varsojen selviytymiseen Helsingin yliopiston Yliopistollisessa hevossairaalassa vuosina 2012–2023.
Tutkimukseen osallistui viisikymmentä alle kuukauden ikäistä keuhkokuumetta sairastavaa varsaa. Varsat jaettiin ryhmiin selviytymisen perusteella (selviytyneet, n=38, ja kuolleet/lopetetut, n=12). Useita kliinisiä ja laboratoriomuuttujia tutkittiin kuoleman riskitekijöinä logistisella regressioanalyysillä yksi- ja monimuuttujamalleilla (ikä, paino, rotu, tamman kantoaika, kliiniset parametrit, tulehdusarvot, veren vastaainetaso sairaalaan tuotaessa, veriviljely, valtimoveren happiosapaine, sairaalahoidon pituus sekä muut mahdolliset diagnoosit mukaan lukien mm. sepsis). Jos muuttujalla oli ennustepotentiaalia varsan selviytymisen suhteen regressioanalyysissä, sen diagnostinen arvo määritettiin receiver operating characteristic (ROC) -käyrän avulla.
Tulokset osoittivat, että yksimuuttuja-analyysissä selviytymisen vetosuhde (OR; 95 % CI) oli negatiivisesti yhteydessä korkeampaan hengitystaajuuteen toisena sairaalahoitopäivänä (1,32; 1,07–1,62; p=0,009) ja positiiviseen veren bakteeriviljelyyn (12,08; 1,88–77,67; p=0,009). Selviytymisen vetosuhde oli positiivisesti yhteydessä lämminveriseen rotuun (0,11; 0,01–0,96; p=0,046) ja pidempään sairaalahoitoon (0,59; 0,40–0,86; p=0,006). Monimuuttuja-analyysissä selviytymisen vetosuhde oli negatiivisesti yhteydessä korkeampaan hengitystaajuuteen toisena sairaalahoitopäivänä (1,36; 1,07–1,71; p=0,011). ROC-analyysissä optimaalinen raja-arvo hengitystaajuudelle oli 55/min (sensitiivisyys 75,0 % ja spesifisyys 76,3 %).
Ennustearvojen perusteella hengitystaajuus <55/min toisena sairaalahoitopäivänä suosi selviytymistä. Muilla tutkituilla muuttujilla ei todettu yhteyttä selviytymiseen. Matalampi hengitystaajuus sairaalahoidon alkuvaiheessa on siis selviytymistodennäköisyyttä lisäävä tekijä keuhkokuumetta sairastavilla varsoilla, sillä korkea hengitystaajuus lisää kuoleman todennäköisyyttä (vetosuhde nousee 36 %). Tutkimuksen havainnot voivat auttaa tunnistamaan jo sairaalahoidon alkuvaiheessa varsoja, joilla on lisääntynyt kuolemanriski. Tämä voi helpottaa hoidon suunnittelua ja ennusteen antamista varsan omistajalle.
Parapoxviruses (PPVs) infect various animal species and humans worldwide. Equine parapoxvirus (EqPPV), first identified in one sick horse in Finland in 2013, was recently recognized as the sixth known PPV species. EqPPV is potentially zoonotic. During the winter of 2021−2022, EqPPV caused a widespread pastern dermatitis outbreak in racehorses in Finland, with earlier reports of similar, unconfirmed outbreaks in 2015 and 2019. This study aimed to develop a serological test for EqPPV and investigate the immune response and seroprevalence of the virus using both clinical and archived equine serum samples. We developed an immunofluorescent assay based on recombinant envelope proteins, and validated it with western blot analysis. The results showed that EqPPV provokes a rapid immune response within days of clinical signs. Two horses were tested one year after a PCR-verified EqPPV infection, and they retained stable IgG titres. Seroprevalence in archived samples peaked in 2015 and 2022, suggesting that EqPPV may have caused a previous outbreak. These findings imply that EqPPV is a re-emerging pathogen capable of causing significant epidemics.
Equine insect bite hypersensitivity (IBH) is an allergic skin disease caused by the allergens in the saliva of Culicoides spp. The IgE-mediated reactions are characterised with chronic seasonal pruritic dermatitis. Pruritis often results to severe skin lesions and significantly impairs the well-being of horses. IBH is thought to be multifactorial; morbidity is influenced by both environmental and genetic factors.
DNA samples were collected from Finnhorses with typical, recurrent (at least 2 consecutive years) IBH symptoms (age minimum 3 years) and healthy controls (age minimum 10 years). The symptoms included pruritus, skin thickening, hair loss and scaling. A case–control genome-wide association study was performed using the Illumina’s Equine 80K Genotyping BeadChip containing 65157 single nucleotide polymorphisms (SNPs). PLINK v1.9 software were used for association analysis by applying the Fisher's exact test and for pruning and quality control.
DNA samples were available from 72 cases (mean age ± SD 11.5 ± 5.9) and 72 controls (mean age ± SD 15.9 ± 4.7). The lowest P-value was found in SNP BIEC2_1080912 (P = 9.241x10-6, OR = 3.719) which is in chromosome 9.
Protein coding gene close to the SNP with the lowest P-value is a potential candidate gene for IBH. Candidate gene has role in the formation and regulation of several immune cells. Additionally, it promotes immune exhaustion. Further studies are required to clarify the role of this candidate gene in IBH in Finnhorses.
