Post-vaccinal distemper-like disease in two dog litters with confirmed infection of vaccine virus strain.

Modified live canine distemper virus (CDV) vaccines are widely used and considered both safe and effective. Although there are occasional literature reports of suspected vaccine-induced disease, there are none where the vaccine strain has been identified in affected tissues. Here we describe two such cases in different litters. In litter A, five of ten puppies presented with fever, anorexia, vomiting, and diarrhea a few days post-vaccination. Four puppies died or were euthanized, and autopsy revealed atypical necrosis of the lymphoid tissue. In litter B, two of five puppies developed typical neurological signs some months post-vaccination and autopsy revealed encephalitis. In all cases, affected organs tested positive for CDV on immunohistochemistry, and CDV RNA extracted from the lesions confirmed the presence of vaccine strain. Since multiple puppies from each litter were affected, it cannot be excluded without further studies that some undiagnosed inherited immunodeficiency disorder may have been involved.

Highly pathogenic avian influenza A(H5N1) virus infection on multiple fur farms in the South and Central Ostrobothnia regions of Finland, July 2023.

Since mid-July 2023, an outbreak caused by highly pathogenic avian influenza A(H5N1) virus clade 2.3.4.4b genotype BB is ongoing among farmed animals in South and Central Ostrobothnia, Finland. Infections in foxes, American minks and raccoon dogs have been confirmed on 20 farms. Genetic analysis suggests introductions from wild birds scavenging for food in farm areas. Investigations point to direct transmission between animals. While no human infections have been detected, control measures are being implemented to limit spread and human exposure.

Becoming confidently competent: a qualitative investigation of training in cognitive functional therapy for persistent low back pain.

BACKGROUND: Physiotherapists trained to deliver biopsychosocial interventions for complex musculoskeletal pain problems often report difficulties in confidence and competency at the end of training. Cognitive Functional Therapy (CFT) is an individualized biopsychosocial intervention and understanding the facilitators and barriers to training in CFT will help inform future training programs. This study aimed to explore physiotherapists' and trainers' perceptions of the process of developing competency in CFT. METHODS: A cross-sectional qualitative design using interviews of 18 physiotherapists and two trainers investigated training in CFT for persistent LBP via reflexive thematic analysis. RESULTS: Physiotherapists reported undergoing a complex behavior change process during training. Four themes emerged: 1) Pre-training factors; 2) Behavior change process; 3) Physiotherapy culture and context; and 4) Confident competence and beyond. Key components included graduated practice exposure linked to experiential learning with feedback and clear competency guidelines. Pre-training and contextual factors were facilitators or barriers depending on the individual. Physiotherapists supported ongoing learning, even after competency was achieved. CONCLUSIONS: This study provides insight into the processes of change during progress toward competency in CFT. It highlights facilitators and barriers to competency including physiotherapy culture and the clinical environment. The study also describes important educational components, including experiential learning and clinical integration, which may be used to inform future post-graduate training.

An adventurous learning journey. Physiotherapists' conceptions of learning and integrating cognitive functional therapy into clinical practice.

Background: Recent low back pain guidelines recommend a BPS approach to the management of disabling low back pain. However, the most effective way of teaching physiotherapists to implement these approaches remains unknown. The present qualitative study aimed to explore physiotherapists' conceptions of learning and integrating Cognitive Functional Therapy (CFT) into clinical practice in Finnish primary health care.Methods: We interviewed 22 physiotherapists, who participated in four to six days of CFT workshops. A phenomenographic approach was used to explore the variation in the physiotherapists' conceptions of this process.Results: Four themes emerged from the data: 1) membership of work community; 2) learning journey; 3) transition to new working methods; and 4) professional role as a physiotherapist. These themes varied in four categories of description: recognizing difference of the new approach, toward integrating the new approach, waking up to explore, commitment to new approach and expanding application of new approach. The critical aspects between the categories that enabled the integration of CFT into clinical practice were ability to overcome resistance and to change views, being shaken and ability to critically reflect on one's work, support from the work community and becoming convinced, creativity, multidisciplinary collaboration and continuous learning.Conclusion: The participants' responses to the training varied greatly, suggesting that for some, the training was insufficient to support adequate changes in their practice behavior; whereas for others, the training was a life changing experience.

Patients' conceptions of undergoing physiotherapy for persistent low back pain delivered in Finnish primary healthcare by physiotherapists who had participated in brief training in cognitive functional therapy.

PURPOSE: To explore the conceptions of patients with persistent low back pain (LBP) of undergoing physiotherapy delivered in Finnish primary healthcare by physiotherapists who had participated in brief training in Cognitive Functional Therapy (CFT). METHODS: As part of a feasibility implementation study exploring CFT in management of LBP in the Finnish primary healthcare system, we interviewed nine patients from four geographical areas in Finland after receiving care. We used a phenomenographic approach to explore the variation in their conceptions. RESULTS: The analysis revealed four descriptive categories: "hung out to dry," "stuck," "making sense and taking control," and "holistic approach to care and living," that varied based on six themes. CONCLUSIONS: Although the participants accepted this approach to care well, there was significant variation in patients' conceptions. Restricted access to care within the healthcare system and a lack of social support led some of them to feel they had been left alone to suffer with their pain. On the other hand, based on the results of this study, positive experiences of physiotherapy and good collaboration with the physiotherapist, wider social support outside of physiotherapy, a better understanding of the multidimensional nature of pain and the acquisition of self-management skills were reported as positive aspects of undergoing physiotherapy that may be related to positive treatment outcomes.Implications for rehabilitationThe participants of this study saw undergoing physiotherapy delivered within biopsychosocial framework as different from their previous physiotherapy experiences and for some the process ended with feeling empty-handed and for others it could be a turning point in their lives.This study encourages the health care providers to create flexible care pathways and ongoing support for more vulnerable individuals so that they don't feel abandoned by the system.Professionals could also pay attention to building strong therapeutic alliance, help patients understand pain in biopsychosocial framework, take into consideration social support networks of the patients, and support patients toward effective self-management strategies.

Training of Physical Therapists to Deliver Individualized Biopsychosocial Interventions to Treat Musculoskeletal Pain Conditions: A Scoping Review.

OBJECTIVE: Current guidelines recommend management of musculoskeletal pain conditions from a biopsychosocial approach; however, biopsychosocial interventions delivered by physical therapists vary considerably in effectiveness. It is unknown whether the differences are explained by the intervention itself, the training and/or competency of physical therapists delivering the intervention, or fidelity of the intervention. The aim was to investigate and map the training, competency assessments, and fidelity checking of individualized biopsychosocial interventions delivered by physical therapists to treat musculoskeletal pain conditions. METHODS: A scoping review methodology was employed, using Arksey and O'Malley's framework. Seven electronic databases were searched between January and March 2019, with a bridge search completed in January 2020. Full-text peer-reviewed articles, with an individualized biopsychosocial intervention were considered, and 35 studies were included. RESULTS: Reporting overall was sparse and highly variable. There was a broad spectrum of training. More sophisticated training involved workshops combining didactic and experiential learning over longer durations with supervision and feedback. Less sophisticated training was brief, involving lectures or seminars, with no supervision or feedback. Competency assessments and fidelity testing were underperformed. CONCLUSIONS: Training in some interventions might not have facilitated the implementation of skills or techniques to enable the paradigm shift and behavior change required for physical therapists to effectively deliver a biopsychosocial intervention. Lack of competency assessments and fidelity checking could have impacted the methodological quality of biopsychosocial interventions. IMPACT: This study highlighted problematic reporting, training, assessment of competency, and fidelity checking of physical therapist-delivered individualized biopsychosocial interventions. Findings here highlight why previous interventions could have shown small effect sizes and point to areas for improvement in future interventions. These findings can help inform future research and facilitate more widespread implementation of physical therapist-delivered biopsychosocial interventions for people with musculoskeletal pain and thereby improve their quality of life.

Effectiveness of training in guideline-oriented biopsychosocial management of low-back pain in occupational health services - a cluster randomized controlled trial.

OBJECTIVE: This study aimed to investigate the effectiveness of brief training in the guideline-oriented biopsychosocial management of low-back pain (LBP) in occupational health services using a cluster-randomized design. A small sample of physiotherapists and physicians from the intervention units (N=12) were given three- to seven-day training focusing on the biopsychosocial management of LBP, while professionals in the control units (N=15) received no such training. METHODS: Eligible patients with LBP, with or without radicular pain, aged 18-65, were invited to participate. A web-based questionnaire was sent to all recruited patients at baseline, three months and one year. The primary outcome measure was disability (Oswestry Disability Index, ODI) over one year. Between-group differences were analyzed using linear and generalized linear mixed models adjusted for baseline-response delay as well as variables showing between-group imbalance at baseline. RESULTS: The final study sample comprised 234 and 81 patients in the intervention and control groups, respectively at baseline, and 137 and 47 patients, respectively, at one year. At baseline, the mean duration of pain was longer in the intervention group (P=0.017), and pain-related fear concerning physical activity was lower (P=0.012). We observed no significant difference between the groups' primary outcome measure (adjusted one-year mean difference in the ODI: 2.3; 95% confidence interval -1.0-5.7; P=0.175) or most secondary outcomes. CONCLUSIONS: Brief training in guideline-oriented biopsychosocial management of LBP for occupational health professionals did not appear to be effective in reducing patients' symptom over one-year follow-up compared to treatment as usual.

Evaluation of training in guideline-oriented biopsychosocial management of low back pain in occupational health services: Protocol of a cluster randomized trial.

BACKGROUND: To prevent low back pain (LBP) from developing into a prolonged disabling condition, clinical guidelines advocate early stage assessment, risk-screening, and tailored interventions. Occupational health services recommend guideline-oriented biopsychosocial screening and individualized assessment and management. However, it is not known whether training a limited number of health care professionals improves the management process. The primary objective of this study is to investigate whether training in the biopsychosocial practice model is effective in reducing disability. Furthermore, we aim to evaluate health-economic impacts of the training intervention in comparison to usual medical care. METHODS: The occupational health service units will be allocated into a training or control arm in a two-arm cluster randomized controlled design. The training of occupational physiotherapists and physicians will include the assessment of pain-related psychosocial factors using the STarT Back Tool and the short version of the Örebro Musculoskeletal Pain Screening Questionnaire, the use of an evidence-based patient education booklet as part of the management of LBP, and tailored individualized management of LBP according to risk stratification. The control units will receive no training. The study population will include patients aged 18-65 with nonspecific LBP. The primary outcome is a patient-reported Oswestry Disability Index from baseline to 12 months. By estimating group differences over time, we aim to evaluate the effectiveness of the training intervention in comparison to usual medical care, and to undertake an economic evaluation using individual patients' health care records (participant-level data) and the participating units' registries (cluster-level data). In addition, through interviews and questionnaires, we will explore the health care professionals' conceptions of the adoption of, the barriers to, and the facilitators of the implementation of the practice model. DISCUSSION: The evaluation of training in the guideline-oriented biopsychosocial management of LBP in occupational health services is justified because LBP represents an enormous burden in terms of work disability.

Susceptibility of rainbow trout to three different genogroups of infectious pancreatic necrosis virus.

Infectious pancreatic necrosis (IPN) is a globally distributed viral disease that is highly prevalent in rainbow trout Oncorhynchus mykiss farms in Finland. Seven genogroups (1-7) of infectious pancreatic necrosis virus (IPNV) exist, of which genogroup 5 (serotype Sp) is generally considered to be the most virulent in European salmonid farming. In Finland, 3 genogroups (2, 5 and 6) have been detected. Genogroup 2 is the most widespread and to date is the only genogroup associated with clinical disease in field observations. A bath challenge model infection trial was conducted to investigate the potential pathogenicity of the existing Finnish IPNV genogroups on IPNV-negative rainbow trout fry. Three Finnish IPNV isolates, a positive control (a Norwegian genogroup 5 isolate previously associated with high virulence in Atlantic salmon Salmo salar) and a negative control were used, and mortality was recorded daily for 8 wk. The Finnish IPNV genogroup 5 isolate caused the highest cumulative mortality, and the genogroup 2 isolate also caused elevated mortalities. The genogroup 6 isolate caused only low mortality, and the positive control treatment showed negligible mortality. Fish exposed to the Finnish genogroup 2 and 5 isolates had IPN-associated lesions, while no lesions were noted in the other treatment groups. These results indicate that Finnish IPNV genogroup 5 is potentially the most virulent IPNV genogroup for Finnish rainbow trout. Interestingly, the Norwegian IPNV genogroup 5 isolate caused only a subclinical IPN infection, providing further evidence for a host species-dependent, virus isolate-related difference in virulence in IPNV genogroup 5. The results also support the continuation of legislative disease control of IPNV genogroup 5 in Finnish inland waters.

Physiotherapists' perceptions of learning and implementing a biopsychosocial intervention to treat musculoskeletal pain conditions: a systematic review and metasynthesis of qualitative studies.

Clinical practice guidelines recommend a biopsychosocial approach for the management of musculoskeletal pain conditions, but physiotherapists have reported feeling inadequately trained and lacking in confidence to deal with psychosocial issues. Although a growing number of studies are exploring physiotherapists' perceptions of biopsychosocial training, the results have not been synthesized. Therefore, the aim of this systematic review and metasynthesis of qualitative studies was to explore physiotherapists' perceptions of learning and implementing a biopsychosocial intervention to treat musculoskeletal pain conditions. A search of the electronic databases: MEDLINE, EMBASE, CINAHL, ERIC, PsycInfo, SportDiscus, and Sociological abstracts identified eligible studies. We included full-text qualitative and mixed-methodology studies published in English, which investigated physiotherapists' perceptions of learning and implementing biopsychosocial interventions. Twelve studies involving 113 participants met the inclusion criteria, and a thematic synthesis was conducted. The quality of the included studies was appraised using the Clinical Appraisal Screening Program. Four main themes emerged from the data: changed understanding and practice, professional benefits, clinical challenges, and learning requirements. The results of this study indicate that although the physiotherapists reported a shift towards more biopsychosocial and person-centered approaches, the training interventions did not sufficiently help them feel confident in delivering all the aspects. Planning future implementation interventions and training physiotherapists through a biopsychosocial approach should focus on adequate training and individualized mentoring related to psychosocial factors, and discussion of role boundaries, patient expectations, and organizational factors such as time constraints and referral pathways.

Association of STarT Back Tool and the short form of the Örebro Musculoskeletal Pain Screening Questionnaire with multidimensional risk factors.

The Short form of the Örebro Musculoskeletal Pain Screening Questionnaire (ÖMPSQ-short) and the STarT Back Tool (SBT) have been developed to screen for risk factors for future low back pain (LBP) -related disability and work loss respectively. The aim of this study was to investigate the accordance of the two questionnaires and to evaluate the accumulation of risk factors in the risk groups of both screening tools in a large population-based sample. The study population consisted of 3079 participants of the Northern Finland Birth Cohort 1966 who had reported LBP over the previous 12 months and had SBT and ÖMPSQ-short data. We evaluated the association of depressive and anxiety symptoms (Hopkins symptom check list-25, Generalized anxiety disorder 7 questionnaire, and Beck's Depression Inventory 21), psychological features (Fear-Avoidance Beliefs Questionnaire), lifestyle characteristics (BMI, smoking, alcohol abuse, physical inactivity) and social factors (education level) with the SBT and ÖMPSQ-short risk groups. The high-risk groups of both questionnaires were associated (p < 0.001) with depressive and anxiety symptoms and fear-avoidance beliefs. In addition, adverse lifestyle factors accumulated in the higher risk groups, especially from the ÖMPSQ-short. Agreement between the two questionnaires was moderate for men and fair for women.

Transcultural adaption and preliminary evaluation of "understanding low back pain" patient education booklet.

BACKGROUND: Low back pain (LBP) is the number one cause of disability globally. LBP is a symptom associated with biological, psychological and social factors, and serious causes for pain are very rare. Unhelpful beliefs about LBP and inappropriate imaging are common. Practitioners report pressure from patients to provide inappropriate imaging. A recently developed patient education and management booklet, 'Understanding low back pain', was designed to target previously identified barriers for reducing inappropriate imaging. The booklet includes evidence-based information on LBP and supports communication between patients and practitioners. Our aim was to 1) describe the translation process into Finnish and 2) study patients' and practitioners' attitudes to the booklet and to evaluate if it improved patients' understanding of LBP and practitioners' ability to follow imaging guidelines. METHODS: We translated the booklet from English to Finnish. Preliminary evaluation of the booklet was obtained from LBP patients (n = 136) and practitioners (n = 32) using web-based questionnaires. Open-ended questions were analysed using thematic analysis. RESULTS: Approximately half of the patients reported that reading the booklet helped them to understand LBP, while a third thought it encouraged them to perform physical activity and decreased LBP-related fear. Eighty percent of practitioners reported that the booklet helped them to follow imaging guidelines. In addition, practitioners reported that they found the booklet helpful and that it decreased the need for imaging. CONCLUSIONS: The booklet seemed to be helpful in LBP management and in decreasing the need for LBP imaging according to patients and practitioners. Further research on the clinical effectiveness of the booklet in controlled study settings is needed. TRIAL REGISTRATION: ISRCTN, ISRCTN14389368, Registered 4 April 2019 - Retrospectively registered; ISRCTN11875357, Registered 22 April 2019 - Retrospectively registered.

An Outbreak of Rabbit Hemorrhagic Disease in Finland.

Rabbit hemorrhagic disease (RHD) was detected in European rabbits ( Oryctolagus cuniculus) for the first time ever in Finland in 2016. Reports of dead feral rabbits in Helsinki started to accumulate from April 2016. The Finnish Food Safety Authority Evira received the first animals in late April, and the main necropsy finding was severe, acute necrotizing hepatitis. Genetic material from RHD virus (RHDV) was detected in the liver and was further characterized as RHDV2. The Finnish virus did not group with RHDV strains from a concurrent outbreak in neighboring Sweden, suggesting another origin. The outbreak peaked in May and lasted until August, after which sightings of both live and dead rabbits became rare. No major outbreaks in domestic rabbits were observed, although infection in one pet rabbit was confirmed.

From "Non-encounters" to autonomic agency. Conceptions of patients with low back pain about their encounters in the health care system.

Low back pain is a considerable health problem which affects people around the world, causing major healthcare costs. The use of qualitative research methods enables us to describe and understand patients' experience of, and attitudes to, healthcare. The aim of the present phenomenographic study was to identify and describe the contextual nature of the conceptions of patients with low back pain of their encounters in the HCS. Seventeen patients with chronic or episodic low back pain classified as "high risk" were interviewed in open recall interviews, using videos of patients' initial physiotherapy sessions that had been recorded previously. The data were analysed using the phenomenographic method. Patients' conceptions of their clinical journey were formulated by a variety of themes: convincing care, lifestyle change, participation, reciprocality and ethicality of encounters. The themes varied in four categories: "non-encounters", seeking support, empowering collaboration and autonomic agency. The results showed a range of clinical interactions - from very negative and disempowering, to empowering and life changing. The key differences between the first and second categories were professionals "being present" and patients starting to understand their low back pain. Between the second and third category, the key aspects were strong therapeutic alliance and the active participation of the patient. Finally, the key differences between the third and fourth categories were the patient being in charge and taking responsibility while knowing that help was available if required. The results may help in improving the care of patients with low back pain.

Molecular characterisation of infectious pancreatic necrosis viruses isolated from farmed fish in Finland.

Infectious pancreatic necrosis virus (IPNV) has been isolated annually since 1987 from salmonids without clinical signs at coastal fish farms in Finland. In the inland area, viral isolations were rare until 2012, when IPNV was detected at several freshwater fish farms. Between 2013 and 2015, the infection spread and IPNV was continuously isolated from several farms, both inland and on the coast. The aim of this study was to genetically characterise the IPNV isolates collected from Finnish coastal and inland fish farms over the last 15 years, and to detect genetic changes that may have occurred in the virus populations during the study period. The partial VP2 gene sequence from 88 isolates was analysed. In addition, a complete genomic coding sequence was obtained from 11 isolates. Based on the genetic analyses, Finnish IPNV isolates belong to three genogroups: 2, 5 and 6. The genetic properties of the isolates appear to vary between inland farms producing juveniles and food fish farms in the coastal region: the inland farms harboured genogroup 2 isolates, whereas at coastal farms, all three genogroups were detected. Little genetic variation was observed within the Finnish genogroup 2 and 5 isolates, whereas among the genogroup 6 isolates, two subgroups were detected. All isolates studied demonstrated amino acid patterns in the viral VP2 gene previously associated with avirulence. However, increased mortality was detected at some of the farms, indicating that more research is needed to clarify the relationship between the pathogenicity and genetic properties of IPNV isolates from different genogroups.

Genome Sequence of a Ranavirus Isolated from Pike-Perch Sander lucioperca.

The pike-perch iridovirus (PPIV) was isolated in Finland from apparently healthy pike-perch fingerlings during routine disease surveillance. Our phylogenomic analysis revealed that PPIV is the first fish member of a clade of ranaviruses previously described from European and Chinese amphibians.

Infectious pancreatic necrosis virus (IPNV) strain with genetic properties associated with low pathogenicity at Finnish fish farms.

Infectious pancreatic necrosis (IPN) is a contagious viral disease of fish that causes economic losses in aquaculture worldwide. In Finland, IPN virus (IPNV) has been isolated since 1987 from adult fish showing no signs of clinical disease at fish farms located in the coastal areas of the Baltic Sea. The inland area of Finland, however, remained free of IPN until 2012, when fish on several rainbow trout farms were diagnosed IPNV-positive. The fish mortalities detected at the farms were low, but clinical signs and histopathological changes typical for IPNV infection were seen in juvenile salmonids. IPNV was isolated at high water temperatures up to 22°C. In 2013 and 2014, IPNV detections continued at inland farms, indicating that infections have spread. The aim of this study was to describe the epidemiology of the outbreak and to characterise the Finnish inland IPNV isolates using histopathological, immunohistochemical and genetic approaches. In order to determine the epidemiological origin of the inland IPNV infections, the partial viral capsid protein (VP2) gene sequences of the inland IPNV isolates were compared with the sequences of the isolates from the coastal farms. Based on the genetic analysis, the inland isolates belong to IPNV Genogroup 2 (Serotype A3/Ab), and the origin of the isolates appears to be one or several coastal fish farms.

Characterization of perch rhabdovirus (PRV) in farmed grayling Thymallus thymallus.

Two Finnish fish farms experienced elevated mortality rates in farmed grayling Thymallus thymallus fry during the summer months, most typically in July. The mortalities occurred during several years and were connected with a few neurological disorders and peritonitis. Virological investigation detected an infection with an unknown rhabdovirus. Based on the entire glycoprotein (G) and partial RNA polymerase (L) gene sequences, the virus was classified as a perch rhabdovirus (PRV). Pairwise comparisons of the G and L gene regions of grayling isolates revealed that all isolates were very closely related, with 99 to 100% nucleotide identity, which suggests the same origin of infection. Phylogenetic analysis demonstrated that they were closely related to the strain isolated from perch Perca fluviatilis and sea trout Salmo trutta trutta caught from the Baltic Sea. The entire G gene sequences revealed that all Finnish grayling isolates, and both the perch and sea trout isolates, were most closely related to a PRV isolated in France in 2004. According to the partial L gene sequences, all of the Finnish grayling isolates were most closely related to the Danish isolate DK5533 from pike. The genetic analysis of entire G gene and partial L gene sequences showed that the Finnish brown trout isolate ka907_87 shared only approximately 67 and 78% identity, respectively, with our grayling isolates. The grayling isolates were also analysed by an immunofluorescence antibody test. This is the first report of a PRV causing disease in grayling in Finland.

Expression analysis of immune response genes in fish epithelial cells following ranavirus infection.

Ranaviruses (family Iridoviridae) are a growing threat to fish and amphibian populations worldwide. The immune response to ranavirus infection has been studied in amphibians, but little is known about the responses elicited in piscine hosts. In this study, the immune response and apoptosis induced by ranaviruses were investigated in fish epithelial cells. Epithelioma papulosum cyprini (EPC) cells were infected with four different viral isolates: epizootic haematopoietic necrosis virus (EHNV), frog virus 3 (FV3), European catfish virus (ECV) and doctor fish virus (DFV). Quantitative real-time PCR (qPCR) assays were developed to measure the mRNA expression of immune response genes during ranavirus infection. The target genes included tumour necrosis factor α (TNF-α), interleukin-1ß (IL-1ß), ß2-microglobulin (ß2M), interleukin-10 (IL-10) and transforming growth factor ß (TGF-ß). All ranaviruses elicited changes in immune gene expression. EHNV and FV3 caused a strong pro-inflammatory response with an increase in the expression of both IL-1ß and TNF-α, whereas ECV and DFV evoked transient up-regulation of regulatory cytokine TGF-ß. Additionally, all viral isolates induced increased ß2M expression as well as apoptosis in the EPC cells. Our results indicate that epithelial cells can serve as an in vitro model for studying the mechanisms of immune response in the piscine host in the first stages of ranavirus infection.

Quantitation of ranaviruses in cell culture and tissue samples.

A quantitative real-time PCR (qPCR) based on a standard curve was developed for detection and quantitation of ranaviruses. The target gene for the qPCR was viral DNA polymerase (DNApol). All ten ranavirus isolates studied (Epizootic haematopoietic necrosis virus, EHNV; European catfish virus, ECV; European sheatfish virus, ESV; Frog virus 3, FV3; Bohle iridovirus, BIV; Doctor fish virus, DFV; Guppy virus 6, GV6; Pike-perch iridovirus, PPIV; Rana esculenta virus Italy 282/I02, REV282/I02 and Short-finned eel ranavirus, SERV) were detected with the qPCR assay. In addition, two fish cell lines - epithelioma papulosum cyprini (EPC) and bluegill fry (BF-2) - were infected with four of the isolates (EHNV, ECV, FV3 and DFV), and the viral quantity was determined from seven time points during the first three days after infection. The qPCR was also used to determine the viral load in tissue samples from pike (Esox lucius) fry challenged experimentally with EHNV.