Control line failure in Angiostrongylus vasorum point-of-care serology test in dogs with angiostrongylosis due to suspected hook effect.

OBJECTIVES: Angiostrongylosis is a significant differential for a diverse range of clinical signs in dogs, many of whom present acutely and sometimes with fatal consequences. Point-of-care diagnostic assays include a commercially available Angiostrongylus vasorum qualitative direct lateral flow assay. MATERIALS AND METHODS: Case records from one referral centre from dogs with an invalid A. vasorum lateral flow assay, comprising an absent control line alongside a visible test line, were reviewed. As control line failure was hypothesised to be due to antigen excess; where available the A. vasorum lateral flow assay was repeated using dilutions of the original serum. RESULTS: Six dogs had an invalid A. vasorum lateral flow assay result. Five dogs had presented with acute-onset, severe clinical disease consistent with angiostrongylosis, and one dog was a clinically healthy in-contact. Clinical suspicion of angiostrongylosis was confirmed using alternative diagnostic testing and/or response to treatment. Repetition of the A. vasorum lateral flow assay, in four cases, using diluted plasma (10% to 12.5% v/v) resulted in the appearance of a control line alongside the visible test line. CLINICAL SIGNIFICANCE: A heavy burden of A. vasorum infection resulting in angiostrongylosis should be suspected in dogs with compatible clinical signs and an invalid A. vasorum lateral flow assay result due to control failure alongside a visible test line. Repetition of the test with a diluted serum may be considered to account for the hook effect, also known as the postzone phenomenon, as a possible cause.

Prevalence and risk factors for common respiratory pathogens within a cohort of pet cats in the UK.

OBJECTIVES: Feline herpesvirus (FHV), feline calicivirus (FCV) and Chlamydia felis are common causes of upper respiratory tract disease (URTD) in cats. Their prevalence in the UK pet cat population has not been reported and little is known regarding the risk factors for their oral carriage. METHODS: Total nucleic acid was extracted from owner-collected buccal swabs (n=600) from cats enrolled in a self-selected longitudinal cohort study. Duplex quantitative PCRs for the detection of FHV and C. felis genomic DNA and reverse-transcriptase quantitative PCRs for the detection of FCV genomic RNA were performed. Duplicates, swabs with insufficient host DNA/RNA, and cats with missing data were excluded. Selected epidemiological data were interrogated using univariable and multi-variable logistic regression modelling to identify risk factors. RESULTS: Data from 430 cats were included in the final statistical model. Of these, 2.1% (n=9/430; 95% CI 1.0% to 3.9%) were positive for FHV, 13.3% (n=57/430; 95% CI 10.2% to 16.8%) positive for FCV and 1.2% (n=5/430; 95% CI 0.4% to 2.7%) positive for C. felis. FCV co-infection was present in five (44%) FHV-positive cats and three (60%) C. felis-positive cats. FCV carriage was more frequent in purebred cats (odds ratio 2.48; 95% CI 1.37 to 4.49) and in cats with current or historical clinical signs compatible with URTD (odds ratio 2.98; 95% CI 1.22 to 7.27). CLINICAL SIGNIFICANCE: FCV was the most frequently encountered URTD pathogen in this sample of cats; this should be noted for disinfectant choice. In cats suspected of having FHV or C. felis infection, assessment for co-infection with FCV is recommended.

Clinical presentation, diagnostic investigations, treatment protocols and outcomes of dogs diagnosed with tick-borne diseases living in the United Kingdom: 76 cases (2005-2019).

OBJECTIVES: To report the presence of tick-borne diseases in dogs living in the United Kingdom. MATERIALS AND METHODS: Dogs with a final diagnosis of tick-borne diseases made between January 2005 and August 2019 at seven referral institutions in the United Kingdom were included in the study. RESULTS: Seventy-six dogs were included: 25 were diagnosed with ehrlichiosis, 23 with babesiosis, eight with Lyme borreliosis and six with anaplasmosis. Fourteen dogs had co-infections with two or three pathogens. Except for those dogs with anaplasmosis and Lyme borreliosis, most dogs with tick-borne diseases had a history of travel to or from endemic countries. However, three dogs with ehrlichiosis, and one dog each infected with Babesia canis and Babesia vulpes did not have any history of travel. A variety of non-specific clinical signs and laboratory abnormalities were reported. Targeted treatment was successful at achieving clinical remission in 64 (84%) dogs. CLINICAL SIGNIFICANCE: Even in non-endemic areas, veterinary surgeons should consider tick-borne diseases in dogs with compatible clinical presentation and laboratory findings and especially where there is a history of travel. As autochthonous transmission of tick-borne-pathogens does occur, an absence of travel should not rule out tick-borne diseases. Specific diagnostic testing is required to confirm infection, and this enables prompt targeted treatment and often a positive outcome.

Adverse childhood experiences and global mental health: avenues to reduce the burden of child and adolescent mental disorders.

Mental disorders are one of the largest contributors to the burden of disease globally, this holds also for children and adolescents, especially in low- and middle-income countries. The prevalence and severity of these disorders are influenced by social determinants, including exposure to adversity. When occurring early in life, these latter events are referred to as adverse childhood experiences (ACEs).In this editorial, we provide an overview of the literature on the role of ACEs as social determinants of mental health through the lenses of global mental health. While the relation between ACEs and mental health has been extensively explored, most research was centred in higher income contexts. We argue that findings from the realm of global mental health should be integrated into that of ACEs, e.g. through preventative and responsive psychosocial interventions for children, adolescents and their caregivers. The field of global mental health should also undertake active efforts to better address ACEs in its initiatives, all with the goal of reducing the burden of mental disorders among children and adolescents globally.

Inflammatory Mediators in the Mesenteric Lymph Nodes, Site of a Possible Intermediate Phase in the Immune Response to Feline Coronavirus and the Pathogenesis of Feline Infectious Peritonitis?

Feline infectious peritonitis (FIP) is an almost invariably fatal feline coronavirus (FCoV)-induced disease thought to arise from a combination of viral mutations and an overexuberant immune response. Natural initial enteric FCoV infection may remain subclinical, or result in mild enteric signs or the development of FIP; cats may also carry the virus systemically with no adverse effect. This study screened mesenteric lymph nodes (MLNs), the presumed first site of FCoV spread from the intestine regardless of viraemia, for changes in the transcription of a panel of innate immune response mediators in response to systemic FCoV infection and with FIP, aiming to identify key pathways triggered by FCoV. Cats with and without FIP, the latter with and without FCoV infection in the MLN, were compared. Higher expression levels in FIP were found for toll-like receptors (TLRs) 2, 4 and 8. These are part of the first line of defence and suggest a response to both viral structural proteins and viral nucleic acid. Expression of genes encoding inflammatory cytokines and chemokines, including interleukin (IL)-1ß, IL-6, IL-15, tumour necrosis factor (TNF)-α, CXCL10, CCL8, interferon (IFN)-α, IFN-ß and IFN-γ, was higher in cats with FIP, consistent with inflammatory pathway activation. Expression of genes encoding transcription factors STAT1 and 2, regulating signalling pathways, particularly of the interferons, was also higher. Among cats without FIP, there were few differences between virus-positive and virus-negative MLNs; however, TLR9 and STAT2 expression were higher with infection, suggesting a direct viral effect. The study provides evidence for TLR involvement in the response to FCoV. This could open up new avenues for therapeutic approaches.

Adjuvant bevacizumab for melanoma patients at high risk of recurrence: survival analysis of the AVAST-M trial.

Background: Bevacizumab is a recombinant humanised monoclonal antibody to vascular endothelial growth factor shown to improve survival in advanced solid cancers. We evaluated the role of adjuvant bevacizumab in melanoma patients at high risk of recurrence. Patients and methods: Patients with resected AJCC stage IIB, IIC and III cutaneous melanoma were randomised to receive either adjuvant bevacizumab (7.5 mg/kg i.v. 3 weekly for 1 year) or standard observation. The primary end point was detection of an 8% difference in 5-year overall survival (OS) rate; secondary end points included disease-free interval (DFI) and distant metastasis-free interval (DMFI). Tumour and blood were analysed for prognostic and predictive markers. Results: Patients (n=1343) recruited between 2007 and 2012 were predominantly stage III (73%), with median age 56 years (range 18-88 years). With 6.4-year median follow-up, 515 (38%) patients had died [254 (38%) bevacizumab; 261 (39%) observation]; 707 (53%) patients had disease recurrence [336 (50%) bevacizumab, 371 (55%) observation]. OS at 5 years was 64% for both groups [hazard ratio (HR) 0.98; 95% confidence interval (CI) 0.82-1.16, P = 0.78). At 5 years, 51% were disease free on bevacizumab versus 45% on observation (HR 0.85; 95% CI 0.74-0.99, P = 0.03), 58% were distant metastasis free on bevacizumab versus 54% on observation (HR 0.91; 95% CI 0.78-1.07, P = 0.25). Forty four percent of 682 melanomas assessed had a BRAFV600 mutation. In the observation arm, BRAF mutant patients had a trend towards poorer OS compared with BRAF wild-type patients (P = 0.06). BRAF mutation positivity trended towards better OS with bevacizumab (P = 0.21). Conclusions: Adjuvant bevacizumab after resection of high-risk melanoma improves DFI, but not OS. BRAF mutation status may predict for poorer OS untreated and potential benefit from bevacizumab. Clinical Trial Information: ISRCTN 81261306; EudraCT Number: 2006-005505-64.

ADHD symptoms and body composition changes in childhood: a longitudinal study evaluating directionality of associations.

BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) is linked to increased risk of overweight/obesity among children and adults. Studies have also implicated obesity as a risk factor for ADHD. However, no studies have evaluated bidirectional, longitudinal associations between childhood fat mass and ADHD symptom severity. OBJECTIVES: We investigate bidirectional associations between ADHD symptoms and measures of body composition between ages 1.5 and 9. We further examine effects of specific eating patterns linked to ADHD on associations between symptom severity and body composition. METHODS: The study utilized data from children (N = 3903) participating in the Generation R cohort (Netherlands). Children were enrolled at birth and retained regardless of ADHD symptoms over time. Cross-lagged and change models examined bidirectional associations between body composition (body mass index/dual-energy X-ray absorptiometry) and ADHD symptoms at four time points in childhood. RESULTS: A child with a clinically concerning ADHD symptom z-score two standard deviations above the mean at age 6 would be expected to experience about 0.22 kg greater fat mass gain measured via dual-energy x-ray absorptiometry between ages 6 and 9, even if they displayed healthy eating patterns (95% CI: 0.11 - 0.28, p < 0.001). Conversely, fat mass at any age did not predict worse ADHD symptoms later. CONCLUSIONS: Beginning in early childhood, more ADHD symptoms predict higher fat mass at later ages. We did not find evidence of a reverse association. Based on these and prior findings, lifestyle counselling during treatment for children with a diagnosis of ADHD should be considered, even if they are diagnosed in early childhood and do not yet have a body mass index of clinical concern.

Epigenetic profiling of ADHD symptoms trajectories: a prospective, methylome-wide study.

Attention-deficit/hyperactivity disorder (ADHD) is a prevalent developmental disorder, associated with a range of long-term impairments. Variation in DNA methylation, an epigenetic mechanism, is implicated in both neurobiological functioning and psychiatric health. However, the potential role of DNA methylation in ADHD symptoms is currently unclear. In this study, we examined data from the Avon Longitudinal Study of Parents and Children (ALSPAC)-specifically the subsample forming the Accessible Resource for Integrated Epigenomics Studies (ARIES)-that includes (1) peripheral measures of DNA methylation (Illumina 450k) at birth (n=817, 49% male) and age 7 (n=892, 50% male) and (2) trajectories of ADHD symptoms (7-15 years). We first employed a genome-wide analysis to test whether DNA methylation at birth associates with later ADHD trajectories; and then followed up at age 7 to investigate the stability of associations across early childhood. We found that DNA methylation at birth differentiated ADHD trajectories across multiple genomic locations, including probes annotated to SKI (involved in neural tube development), ZNF544 (previously implicated in ADHD), ST3GAL3 (linked to intellectual disability) and PEX2 (related to perixosomal processes). None of these probes maintained an association with ADHD trajectories at age 7. Findings lend novel insights into the epigenetic landscape of ADHD symptoms, highlighting the potential importance of DNA methylation variation in genes related to neurodevelopmental and peroxisomal processes that play a key role in the maturation and stability of cortical circuits.

DNA methylation and substance-use risk: a prospective, genome-wide study spanning gestation to adolescence.

Epigenetic processes have been implicated in addiction; yet, it remains unclear whether these represent a risk factor and/or a consequence of substance use. Here, we believe we conducted the first genome-wide, longitudinal study to investigate whether DNA methylation patterns in early life prospectively associate with substance use in adolescence. The sample comprised of 244 youth (51% female) from the Avon Longitudinal Study of Parents and Children (ALSPAC), with repeated assessments of DNA methylation (Illumina 450k array; cord blood at birth, whole blood at age 7) and substance use (tobacco, alcohol and cannabis use; age 14-18). We found that, at birth, epigenetic variation across a tightly interconnected genetic network (n=65 loci; q<0.05) associated with greater levels of substance use during adolescence, as well as an earlier age of onset amongst users. Associations were specific to the neonatal period and not observed at age 7. Key annotated genes included PACSIN1, NEUROD4 and NTRK2, implicated in neurodevelopmental processes. Several of the identified loci were associated with known methylation quantitative trait loci, and consequently likely to be under significant genetic control. Collectively, these 65 loci were also found to partially mediate the effect of prenatal maternal tobacco smoking on adolescent substance use. Together, findings lend novel insights into epigenetic correlates of substance use, highlight birth as a potentially sensitive window of biological vulnerability and provide preliminary evidence of an indirect epigenetic pathway linking prenatal tobacco exposure and adolescent substance use.

Degenerative Encephalopathy in Nova Scotia Duck Tolling Retrievers Presenting with a Rapid Eye Movement Sleep Behavior Disorder.

BACKGROUND: Neurodegenerative diseases are a heterogeneous group of disorders characterized by loss of neurons and are commonly associated with a genetic mutation. HYPOTHESIS/OBJECTIVES: To characterize the clinical and histopathological features of a novel degenerative neurological disease affecting the brain of young adult Nova Scotia Duck Tolling Retrievers (NSDTRs). ANIMALS: Nine, young adult, related NSDTRs were evaluated for neurological dysfunction and rapid eye movement sleep behavior disorder. METHODS: Case series review. RESULTS: Clinical signs of neurological dysfunction began between 2 months and 5 years of age and were progressive in nature. They were characterized by episodes of marked movements during sleep, increased anxiety, noise phobia, and gait abnormalities. Magnetic resonance imaging documented symmetrical, progressively increasing, T2-weighted image intensity, predominantly within the caudate nuclei, consistent with necrosis secondary to gray matter degeneration. Abnormalities were not detected on clinicopathological analysis of blood and cerebrospinal fluid, infectious disease screening or urine metabolite screening in most cases. Postmortem examination of brain tissue identified symmetrical malacia of the caudate nuclei and axonal dystrophy within the brainstem and spinal cord. Genealogical analysis supports an autosomal recessive mode of inheritance. CONCLUSIONS AND CLINICAL IMPORTANCE: A degenerative encephalopathy was identified in young adult NSDTRs consistent with a hereditary disease. The prognosis is guarded due to the progressive nature of the disease, which is minimally responsive to empirical treatment.

Maternal depression symptoms, unhealthy diet and child emotional-behavioural dysregulation.

BACKGROUND: Maternal depression and unhealthy diet are well-known risk factors for adverse child emotional-behavioural outcomes, but their developmental relationships during the prenatal and postnatal periods are largely uncharted. This study sought to examine the inter-relationships between maternal depression symptoms and unhealthy diet (assessed during pregnancy and postnatal periods) in relation to child emotional-behavioural dysregulation (assessed at the ages of 2, 4 and 7 years). METHOD: In a large prospective birth cohort of 7814 mother-child pairs, path analysis was used to examine the independent and inter-related associations of maternal depression symptoms and unhealthy diet with child dysregulation. RESULTS: Higher prenatal maternal depression symptoms were prospectively associated with higher unhealthy diet, both during pregnancy and the postnatal period, which, in turn, was associated with higher child dysregulation up to the age of 7 years. In addition, during pregnancy, higher maternal depression symptoms and unhealthy diet were each independently associated with higher child dysregulation up to the age of 7 years. These results were robust to other prenatal, perinatal and postnatal confounders (such as parity and birth complications, poverty, maternal education, etc.). CONCLUSIONS: Maternal depression symptoms and unhealthy diet show important developmental associations, but are also independent risk factors for abnormal child development.

Environmental risk, Oxytocin Receptor Gene (OXTR) methylation and youth callous-unemotional traits: a 13-year longitudinal study.

Youth with high callous-unemotional traits (CU) are at risk for early-onset and persistent conduct problems. Research suggests that there may be different developmental pathways to CU (genetic/constitutional vs environmental), and that the absence or presence of co-occurring internalizing problems is a key marker. However, it is unclear whether such a distinction is valid. Intermediate phenotypes such as DNA methylation, an epigenetic modification regulating gene expression, may help to clarify etiological pathways. This is the first study to examine prospective inter-relationships between environmental risk (prenatal/postnatal) and DNA methylation (birth, age 7 and 9) in the prediction of CU (age 13), for youth low vs high in internalizing problems. We focused on DNA methylation in the vicinity of the oxytocin receptor (OXTR) gene as it has been previously implicated in CU. Participants were 84 youth with early-onset and persistent conduct problems drawn from the Avon Longitudinal Study of Parents and Children. For youth with low internalizing problems (46%), we found that (i) OXTR methylation at birth associated with higher CU (age 13) as well as decreased experience of victimization during childhood (evocative epigenetic-environment correlation; birth-age 7), (ii) higher prenatal parental risks (maternal psychopathology, criminal behaviors, substance use) associated with higher OXTR methylation at birth and (iii) OXTR methylation levels were more stable across time (birth-age 9). In contrast, for youth with high internalizing problems, CU were associated with prenatal risks of an interpersonal nature (that is, intimate partner violence, family conflict) but not OXTR methylation. Findings support the existence of distinct developmental pathways to CU.

Feline blood genotyping versus phenotyping, and detection of non-AB blood type incompatibilities in UK cats.

OBJECTIVES: The aim of this study was to determine the agreement between AB blood phenotyping and genotyping and determine whether non-AB blood type incompatibilities exist in UK cats. METHODS: Blood samples underwent phenotyping (A, B or AB) using microplate agglutination, and genotyping (AA, Ab or bb) using pyrosequencing of a fragment of the cytidine monophospho-N-acetylneuraminic acid hydroxylase gene. Non-AB blood type incompatibilities were investigated by cross-matching against reference blood of the same phenotype. RESULTS: Of 112 cats tested, 86 (77%) were blood phenotype A, 19 (17%) type B and 7 (6%) type AB. Genotype and initial phenotype agreed in 96% (107 of 112) of cats, but 5 were discordant; these were all B phenotype with either AA (n=2) or Ab (n=3) genotype. Two of the five cats had repeat blood samples tested: one was reclassified as phenotype A; the other remained phenotype B. Two cats had incompatibilities on minor cross-match, but these were attributed to phenotyping errors. CLINICAL SIGNIFICANCE: Unknown mutation(s) associated with phenotype B, resulting in false AA or Ab genotyping, were evident in a small number of cases in this study. No conclusive evidence for non-AB blood type incompatibilities was found.

Non-ribosomal phylogenetic exploration of Mollicute species: new insights into haemoplasma taxonomy.

Nine species of uncultivable haemoplasmas and several Mycoplasma species were examined by partial sequencing of two protein-encoding housekeeping genes. Partial glyceraldehyde-3-phosphate dehydrogenase (gapA) and heat shock protein 70 (dnaK) gene sequences were determined for these Mollicute species; in total nine gapA sequences and ten dnaK sequences were obtained. Phylogenetic analyses of these sequences, along with those of a broad selection of Mollicute species downloaded from GenBank, for the individual genes, and for the gapA and dnaK concatenated data set, revealed a clear separation of the haemoplasmas from other species within the Mycoplasma genus; indeed the haemoplasmas resided within a single clade which was phylogenetically detached from the pneumoniae group of Mycoplasmas. This is the first report to examine the use of gapA and dnaK, as well as a concatenated data set, for phylogenetic analysis of the haemoplasmas and other Mollicute species. These results demonstrate a distinct phylogenetic separation between the haemoplasmas and Mycoplasmas that corresponds with the biological differences observed in these species, indicating that further evaluation of the haemoplasmas' relationship with the Mycoplasma genus is required to determine whether reclassification of the haemoplasmas is necessary.

Developing effective health and safety training materials for workers in beryllium-using industries.

OBJECTIVE: Despite reduced workplace exposures, beryllium sensitization and chronic beryllium disease still occur. Effective health and safety training is needed. METHODS: Through an Occupational Safety and Health Administration (OSHA) Targeted Topic Training grant and company partners, we developed a training program. Evaluation and validation included knowledge and training reaction assessments and training impact survey. RESULTS: We describe herein the iterative, five-pronged approach: (1) needs assessment; (2) materials development; (3) pilot-testing, evaluation, and material revisions; (4) worker training; and (5) evaluation and validation. Mean posttraining test score increased 14% (82% to 96%; P < 0.005) and were unchanged at 90-day follow-up (94%; P = 0.744). In addition, 49% reported making changes in work practices. CONCLUSIONS: The use of a five-pronged training program was effective and well received and resulted in improved work practices. These materials are available on the OSHA Web site.

Haemoplasmas: lessons learnt from cats.

The haemotropic mycoplasmas (haemoplasmas) are a group of bacteria that can induce anaemia in a wide variety of mammals, including domestic cats and wild felids. Different feline haemoplasma species of varying pathogenicity exist, with the more pathogenic Mycoplasma haemofelis (Mhf) capable of inducing severe haemolytic anaemia, whilst 'Candidatus Mycoplasma haemominutum' (CMhm) and 'Candidatus Mycoplasma turicensis' (CMt) are infrequently associated with clinical disease. Chronic haemoplasma infections are common and cats are frequently infected by two or more haemoplasmas, complicating the clinical picture. The natural route of transmission of haemoplasma infection between cats has not yet been determined; however, experimental transmission has been demonstrated via both oral and parenteral administration of infected blood. To date the haemoplasmas have been unable to be cultured in vitro, and accurate diagnosis is currently reliant on detection of bacterial DNA using PCR assays. Treatment of clinical haemoplasmosis is focussed on supportive care in combination with empirical treatment with antimicrobials (tetracyclines or fluoroquinolones). A significant number of asymptomatic cats are positive for haemoplasma infection. These cats may play a role in the maintenance of haemoplasma infection within a population, and need to be considered when choosing potential blood donors. Use of PCR assays has provided an accurate method of diagnosing haemoplasma infection and quantifying response to therapy, including in non-feline host animals, as presumed zoonotic haemoplasma infections are now being documented. Recent advances in genome sequencing techniques have allowed the whole genome sequences of the feline haemoplasmas Mhf and CMhm to be derived, as well as a number of non-feline haemoplasma species. These data have aided the identification of antigens for use in the development of serological tests, allowed the proteomic study of haemoplasmas and provided clues as to how the haemoplasmas can persist within the host. Future areas of study include investigation of their zoonotic potential, mechanisms of immune system evasion and transmission of these emerging pathogens.

A parsimonious model of lineage-specific expansion of MADS-box genes in Physcomitrella patens.

KEY MESSAGE: The MADS-box gene family expanded in the lineage leading to the moss, Physcomitrella patens , mainly as a result of polyploidisations and/or large-scale segmental duplication events and to a lesser extent by tandem duplications. Plant MADS-box genes comprise a large family best known for the roles of type II MIKC (C) genes in floral organogenesis, but also including type II MIKC* genes, some of which have been implicated in male gametophytic development, and type I genes, a few of which are involved in ontogeny of female gametophytes, seeds and embryos. Genome-wide analyses of the MADS-box family in angiosperms have revealed numeric predominance of type I and MIKC (C) genes and cross-species phylogenetic clustering of the Mα, Mß and Mγ subtypes of type I genes and of 12 major subgroups of MIKC (C) genes. The genome sequence of Physcomitrella patens has facilitated investigation of its full complement of 26 MADS-box genes, including 6 MIKC (C) genes, 11 MIKC* genes, seven type I genes and two pseudogenes. A much higher degree of similarity in sequence and architecture within the MIKC (C) and MIKC* gene subtypes exists in Physcomitrella than in Arabidopsis. Furthermore, MADS-box and K-box sequence is highly conserved between the MIKC (C) and MIKC* subgroups in Physcomitrella. Nine MIKC* genes and two MIKC (C) genes are located in pairs or triplets on individual DNA scaffolds. Phylogenetic gene clustering, gene architectures and gene linkages (directly determined from examination of the genome sequence) underpin a parsimonious model of two tandem duplications and three segmental duplication events, which can account for lineage-specific expansion of the MADS-box gene family in Physcomitrella from 4 members to 26. Two of these segmental duplication events may be indicative of polyploidisations, one of which has been postulated previously.

Phylogenetic analysis of feline coronavirus strains in an epizootic outbreak of feline infectious peritonitis.

BACKGROUND: Feline coronavirus (FCoV) infection is common. In a small percentage of cats, FCoV infection is associated with the fatal disease feline infectious peritonitis (FIP). Genetically distinct virulent and avirulent strains of FCoV might coexist within a cat population. OBJECTIVES: To determine whether the strains of FCoV in FIP-affected cats are closely related or genetically distinct from the fecally derived strains of FCoV in contemporary-asymptomatic cats during an epizootic outbreak of FIP. ANIMALS: Four cats euthanized because of FIP and 16 asymptomatic cats. METHODS: This prospective outbreak investigation was initiated during an outbreak of FIP in cats within or rehomed from a rescue/rehoming center. Postmortem samples were collected from cats with FIP and contemporaneous fecal samples from asymptomatic cats. RNA was purified from tissue and fecal samples, FCoV gene fragments were reverse transcribed, PCR-amplified using novel primers, and sequenced. Sequences were aligned with ClustalW and compared with published FCoV sequences. RESULTS: FCoV RNA was detected in all 4 FIP cat postmortem samples and in 9 of the 16 fecal samples from contemporary-asymptomatic cats. Novel primers successfully amplified fragments from 4 regions of the genome for all FCoV-positive samples. Phylogenetic analysis showed that the FIP-associated strains of FCoV from the outbreak were very closely related to the fecally derived strains of FCoV from contemporary-asymptomatic cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Sequence analysis provided no evidence that genetically distinct virulent and avirulent strains of FCoV were present during this FIP outbreak.