Prevalence of selected cardiotropic pathogens in the myocardium of adult dogs with unexplained myocardial and rhythm disorders or with congenital heart disease.

OBJECTIVE: To determine the prevalence of nucleic acid from selected cardiotropic pathogens in endomyocardial biopsy samples from dogs with unexplained myocardial and rhythm disorders (UMRD) and compare prevalence with that for a group of control dogs with congenital heart disease (CHD). ANIMALS: 47 client-owned dogs. PROCEDURES: Right ventricular endomyocardial biopsy was performed in dogs with UMRD (dilated cardiomyopathy [n = 25], atrioventricular block [6], and nonfamilial ventricular [4] and supraventricular arrhythmias [2]) or CHD (10) that required right ventricular catheterization. Biopsy samples were evaluated histologically, and PCR assays were used for detection of nucleic acid from 12 pathogens. RESULTS: 197 biopsy samples were collected from dogs with UMRD (n = 172) or CHD (25). At least 1 pathogen was detected in 21 of 37 (57%; 95% confidence interval [CI], 41% to 71%) dogs with UMRD, and canine coronavirus was detected in 1 of 10 (10%; 95% CI, 2% to 40%) dogs with CHD. Dogs with UMRD were significantly more likely than dogs with CHD to have pathogens detected in biopsy samples (OR, 11.8; 95% CI, 1.3 to 103.0). The most common pathogens in dogs with UMRD were canine distemper virus, canine coronavirus, canine parvovirus 2, and Bartonella spp. No pathogens were detected in available blood samples from dogs with pathogens detected in biopsy samples. CONCLUSIONS AND CLINICAL RELEVANCE: Detection of nucleic acids from selected cardiotropic pathogens in myocardial tissue from dogs with UMRD suggested a possible association between the 2. Further studies are needed to explore whether this association is causative or clinically important. (J Am Vet Med Assoc 2019;255:1150-1160).

Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in small rodents and attached ticks in the Northern Apennines, Italy.

Ticks and ear biopsies were collected from wild small rodents in 2011 and 2012 in the northern Apennines (Italy), up to 1650 m above sea level. Apodemus spp. (n = 83) and Myodes glareolus (n = 22) were infested by Ixodes ricinus (192 larvae and two nymphs), Dermacentor marginatus (179 larvae and 29 nymphs), and Ixodes trianguliceps (three larvae and two nymphs). We detected several Borrelia burgdorferi sensu lato (s.l.) genospecies (B. afzelii, B. burgdorferi sensu stricto, B. garinii, B. lusitaniae, B. valaisiana) in I. ricinus and skin biopsies. The most common genospecies found in I. ricinus was B. valaisiana, while it was B. lusitaniae in tissues. Spotted fever group (SFG) rickettsiae (Rickettsia monacensis, R. slovaca and R. raoultii) infected I. ricinus, D. marginatus and rodent tissues. Rickettsia slovaca was the Rickettsia species most frequently found in our samples. Coinfections by B. burgdorferi s.l. and SFG rickettsiae indicate an overlap of transmission cycles and potential risk for humans to be infected by multiple pathogens, resulting in more severe symptoms. The findings of B. lusitaniae and R. slovaca in bank voles, and of B. valaisiana in small rodents, open new questions about host-pathogen interactions. In addition, our results highlight the importance of small rodents as data sources for studying tick-borne pathogens.

Pectin-honey hydrogel: Characterization, antimicrobial activity and biocompatibility.

BACKGROUND: Novel pectin-honey hydrogels have been developed and characterized as medical device. Ideally, a wound dressing should maintain optimal fluid affinity, permit moisture evaporation, protect the wound from microbes, and have shape-conformability, biocompatibility, and antibacterial activity. OBJECTIVE: A novel, simple and fast method to produce pectin-honey wound dressings is described. METHODS: The properties of these pectin-honey hydrogels were investigated, including swelling ability, water vapour transmission rate, hydrogen peroxide production, methylglyoxal content and antibacterial activity. Biocompatibility was assessed by proliferation assays using cultured fibroblast cells and by in vivo study with subcutaneous and intraperitoneal implantation in rats. RESULTS: Hydrogel showed a good water vapour transmission rate, fluid uptake and were not cytotoxic for fibroblasts. The hydrogel demonstrated good antibacterial activity toward clinically relevant pathogens, including S. aureus and E. coli. Biocompatibility was confirmed by the measurement of plasma levels of interleukin (IL)1 beta, IL-6, tumour necrosis factor (TNF) alpha, and prostaglandin (PG)E2. No histological changes were observed. CONCLUSIONS: The presence of a natural active component, conformability, and complete resorbability are the main characteristics of this new biocompatible biomaterial that is well tolerated by the body, possibly improves healing, may be used for surgical complications prevention, with a simple and inexpensive production process.

How Behavioral Changes Can Indicate Serious Cerebral Pathology: A Case Report of Concomitant Olfactory Neuroblastoma and Distemper Virus Encephalitis in a Swiss Shepherd Dog.

Behavioral alterations in dogs are not easy to understand and cure. The situation is more difficult when an encephalitis due to Canine Distemper Virus (CDV) and a concomitant olfactory neuroblastoma are present. This case report deals with the story of a 5-year-old Swiss shepherd dog with behavioral changes, seizures, epistaxis and ataxia. Following clinical and laboratory exams, a suspected diagnosis of CDV infection was hypothesized, and a therapy based on Ω-interferon was administered. Every supporting therapy failed and the worsening of the clinical conditions led to the euthanasia of the patient. A neoformation in the right frontal lobe was found post mortem. Histopathology and immunohistochemistry investigation showed a non-suppurative demyelinating encephalitis, suggestive of CDV infection, and a desmoplastic epithelioid olfactory neuroblastoma. To the best of authors' knowledge, this is the first clinical pathological report of a non-suppurative encephalitis due to CDV infection and olfactory neuroblastoma in a dog.

Pro-inflammatory cytokines and structural biomarkers are effective to categorize osteoarthritis phenotype and progression in Standardbred racehorses over five years of racing career.

BACKGROUND: Joint impact injuries initiate a progressive articular damage finally leading to post-traumatic osteoarthritis (PTOA). Racehorses represent an ideal, naturally available, animal model of the disease. Standardbred racehorses developing traumatic osteoarthritis of the fetlock joint during the first year of their career were enrolled in our study. Age-matched controls were contemporarily included. Biomarker levels of equine osteoarthritis were measured in serum and synovial fluid (SF) at baseline, and repeated yearly over the next 4 years of training (from T1 to T4). The effect of time and disease on the biomarker concentrations were analysed, and their relationship with clinical and radiographic parameters were assessed. We hypothesized that the kinetics of pro-inflammatory cytokines and structural biomarkers of joint disease would demonstrate progression of degenerative joint status during post-traumatic osteoarthritis and clarify the effect of early joint trauma. RESULTS: The concentrations of IL1-ß, IL-6, TNF-α in the SF of PTOA group peaked at T0, decreased at T1, and then progressively increased with time, reaching levels higher than those observed at baseline starting from T3. CTXII and COMP levels were similar in PTOA and control horses at baseline, and increased in serum and synovial fluid of PTOA horses starting from T2 (serum and synovial CTXII, and serum COMP) or T3 (synovial COMP). The percentual change of TNF-α in the SF of the affected joints independently contributed to explaining the radiological changes at T3 vs T2 and T4 vs T3. CONCLUSIONS: Temporal changes of selected biomarkers in STBRs with an acute episode of traumatic fetlock OA demonstrated that long-term increased concentrations of inflammatory cytokines, type II collagen fragments and COMP, in the SF and serum, are related to PTOA. Based on the observed decrease in inflammatory merkers at T1, we hypothesize that the progression of PTOA could be effectively modulated by proper treatment strategies. Annual variations of synovial concentration of TNF-α can reliably predict radiographic progression of PTOA.

Molecular analysis and associated pathology of beak and feather disease virus isolated in Italy from young Congo African grey parrots (Psittacus erithacus) with an "atypical peracute form" of the disease.

This study is the first report on the genetic and pathogenic characterization of beak and feather disease virus (BFDV) occurring in Italy. Twenty BFDV strains isolated in Italy from juvenile Congo African grey parrots (Psittacus erithacus) were investigated. Seventeen strains showed an "atypical peracute form" (aPF) of the disease, and three a chronic form (CF). The birds with aPF had been weaned, were independent as far as food and protection were concerned and apparently were without lesions. The gene coding for the putative coat protein was amplified in all isolates while the BFDV genome was sequenced completely in 10 samples, eight of them belonging to aPF affected birds and two from CF of the disease. All full genomes clustered into the J strain of BFDV, where two new subtypes were identified. Recombination analyses showed evidence of genetic exchanges in two BFDV genomes. In addition, a correlation between viral isolate and origin of the breeding material was shown, while an association between the genetic features of the virus and the clinical form was not observed. Histologically, apoptosis was detected frequently in aPF samples and sporadically in CF samples. Interestingly, BFDV antigens were detected in the nuclei and cytoplasm of such apoptotic cells. The data presented here support the hypothesis that, in the absence of a defined BFDV genetic variant accountable for a specific clinical form of psittacine beak and feather disease, differences in the apoptotic rate between aPF and CF are strictly host related.

Colostrum and milk can transmit jaagsiekte retrovirus to lambs.

Ovine pulmonary adenocarcinoma (OPA) is a contagious disease caused by jaagsiekte sheep retrovirus (JSRV). In the three studies performed, we have obtained data of the importance of colostrum/milk (C/M) in the transmission of JSRV. In the first study, a group of sheep from a flock with a long history of OPA, samples from colostrum and peripheral blood leucocytes (PBLs) were collected. Two specific PCRs (U3-LTR and env of the JSRV) were carried out. Using U3PCR 8/34 sheep were positive in colostrum whereas with envPCR 7/34 were positive. From these animals only one was positive with U3PCR in the PBLs. Evidence of the transmission of JSRV infection by C/M was obtained in two more separate studies. In the second study, PBLs from five lambs from JSRV+ ewes and two from JSRV-ewes were tested by the U3PCR. They were fed C/M by their mothers during 3 months and slaughtered 7 months after birth. Three out of five lambs from the JSRV+ sheep become PBL positive at 3-4 months old and the other two were also positive at 4-6 months of age. One lamb of the JSRV-sheep became also PBL positive at an age of 3 months. In the third study, a group of lambs from JSRV negative mothers were fed with C/M from JSRV+ sheep and housed in separate unit. For comparison, another group of the same origin and maintained in another different unit, were fed with C/M containing a JSRV virus preparation. All lambs were blood sampled monthly and JSRV infection was detected as early as 15 days and several times onwards in both groups. Control groups fed with C/M from JSRV free flock and JSRV blood test negative sheep were always negative. Together these results indicate that suckling is an important natural transmission route for JSRV.

Genetic characterization of small ruminant lentivirus in Italian mixed flocks: evidence for a novel genotype circulating in a local goat population.

In this study, characterization of the gag gene of small ruminant lentiviruses was carried out in Italian mixed flocks. The nearly complete gag gene was amplified and sequenced. Within genotype A, subtype A1 and a novel subtype, A8, were found in goats, and another novel subtype, A9, was found in both sheep and goats. Subtype B1 was found in both host species and subtype B2 was identified only in sheep. A novel, highly divergent sequence was obtained from goats in two epidemiologically related flocks and is proposed to represent a novel genotype, E. Major epitopes of matrix and capsid antigen were highly divergent, suggesting that serological identification of animals infected with genotype E may have been missed by using currently available diagnostic tests. A recombinant subunit ELISA, based on genotype E-specific epitopes, was developed and a third independent flock carrying this genotype was identified, based on serology.

Antibody response in sheep experimentally infected with different small ruminant lentivirus genotypes.

Two groups of sheep were experimentally infected by intratracheal route with two small ruminant lentivirus (SRLV) isolates belonging to different genotypes (It-561 genotype A3 and It-Pi1 genotype B2). Seroconversion was evaluated using recombinant homologous and heterologous matrix protein/capsid antigen fusion protein. Results clearly indicate that seroconversion against homologous antigen was detected well in advance as regards heterologous antigen in both groups, although the advantage of using homologous antigen was less evident in detecting seroconversion against the caprine arthritis encephalitis virus (CAEV)-like strain, compared with the maedi-visna virus (MVV)-like infection. Commercially available ELISAs detect CAEV-like seroconversion earlier than MVV-like infection suggesting a closer relationship between CAEV-like isolate and the antigen used in the latter ELISA tests. Seven recombinant subunits developed from matrix protein and capsid antigen of strain K1514 (prototype A1) were used to better define the antibody response in sheep infected with It-561 isolate. Two animals clearly reacted against type specific epitopes in the early stage of infection. This study highlights the relative insensitivity of gag encoded cross-reacting epitopes during the early stage of infection and suggests the development of novel diagnostic tests based on both genotype specific antigens.

Relevance of Akt phosphorylation in cell transformation induced by Jaagsiekte sheep retrovirus.

Expression of the JSRV envelope (Env) is sufficient to transform immortalized rodent fibroblasts. A putative docking site for the PI3-K kinase (Y(590)-X-X-M(593)) in the cytoplasmic tail of the transmembrane domain of the JSRV Env is a major determinant of viral-induced cell transformation. Akt is constitutively phosphorylated in rodent fibroblasts transformed by the JSRV Env. However, recent data suggest that Y590 and M593 are not necessary for JSRV Env-induced transformation of the immortalized chicken fibroblasts cell line DF-1. In this study we found that JSRV-induced transformation of DF-1 cells is Akt-independent. In addition, a replication-competent avian vector expressing the JSRV Env (RCASBP(A)+JE) was also able to induce transformation of primary chicken embryo fibroblasts (CEF). Vectors expressing JSRV Env Y590 mutants were still able to induce CEF cells transformation but not as efficiently as the vectors expressing the wild-type Env. In CEF cells, as in DF-1 cells, only the expression of the wild-type Env induced constitutive phosphorylation of Akt. Thus, in chicken cells, the degree of transformation induced by the JSRV Env is maximum in the presence of Y590 and Akt phosphorylation. We addressed the significance of Akt phosphorylation in rat 208F cells transformed by the JSRV Env and showed that Akt is indeed activated and shows kinase activity. Inhibitors of the PI-3K/Akt pathway reproducibly decreased the transformation efficiency of the JSRV Env. In vivo, we found phosphorylated Akt only in nasal tumors induced by the enzootic nasal tumor virus (ENTV), a JSRV-related beta-retrovirus. No evidence of Akt phosphorylation was found in lung tumor sections of sheep affected by pulmonary adenocarcinoma. As a whole, these results suggest that the activation of the PI-3K/Akt pathway contributes to the process of JSRV-induced cell transformation but most likely is not the primary determinant both in vitro and in vivo.