Distemper In Dogs Research Articles

Prevalence of canine distemper in dogs referred to Veterinary Hospital of Ferdowsi University of Mashhad, Mashhad, Iran.

Canine distemper virus (CDV) is responsible for high morbidity and mortality in dogs worldwide. Epidemiological study of canine distemper can help to control and treat the disease in any area. This study aimed to investigate the prevalence of CDV in dogs referred to the Veterinary Hospital from September 23, 2018 to September 22, 2019. Dogs with at least two clinical signs of canine distemper underwent blood tests, rapid test kit from the eye and cerebrospinal fluid (CSF), and RT-PCR from whole blood and/or CSF samples. Out of 1212 referred dogs, 112 dogs were suspected to have canine distemper of which 90 underwent RT-PCR and rapid test kits. The disease prevalence was 4.04% (49/1212) and 7.44% (49/659) according to the total number of referring dogs and number of referring sick dogs, respectively. The distemper fatality rate was 69.57% (32/46). Seventy percent of distemper positive cases were under 12 months old and 52.08% were under 6 months old. Female dogs were more susceptible than males; however, the fatality rate of males was more than females. Of distemper positive dogs, 91.84% were unvaccinated. The highest prevalence (71.43%) of dogs diagnosed with CDV occurred during the cold seasons. It is concluded that canine distemper is endemic in the geographical area of Mashhad and its prevalence rate in dogs referred to the Veterinary Hospital of Ferdowsi University of Mashhad is 4.04% and its fatality rate is 69.57%. This indicates that a significant number of dogs may die if they develop distemper despite treatment.

Некоторые аспекты эпизоотологии, патогенеза и лечения парвовирусного энтерита собак

The nosoprofile of infectious pathology in dogs is widespread in the conditions of the Klintsovskaya veterinary hospital. The most common disease was bordetellosis, which tended to decrease. Draws attention to the susceptibility of dogs to parvovirus enteritis, the incidence of which was consistently high for 3 years (22,4–23,2 %). The lowest rate of diseased animals was observed from such viral infections as dog distemper, coronavirus enteritis and rotavirus infection. In the age aspect, the nosoprofile of parvovirus enteritis is more common in young dogs under 1 year old, they account for 69,6 % of all cases. In seasonal dynamics, the disease is most often recorded in the spring-summer period. In the blood of dogs with parvovirus enteritis, the morphological composition was characterized by a decrease in erythrocytes and leukocytes, but there was an increase in hemoglobin concentration. On the 1st and 8th days of the experiment, there was a decrease in erythrocytes compared to the control by 47,7 and 33,2 %, respectively, and the content of leukocytes in this period was lower than the control values by 2,6 and 1,3 times. The hemoglobin concentration was higher than the control analogues by 37,3 and 20,8 %. The leukoformula showed an increase in stab neutrophils and lymphocytes. In the blood of dogs with parvovirus enteritis, a decrease in the content of total protein, glucose and cholesterol was recorded. Thus, the total protein on the 1st and 8th days tended to decrease throughout the experiment by 19,1 and 28,1 %, respectively. Glucose had a downward trend, this was especially noticeable on the 8th day of the study (33,3 %). The decrease in cholesterol was most marked on the 1st day of the experiment (31,1 %). The applied complex therapy using the probiotic lactobifadol forte contributed to the normalization of the clinical status on the 7th day, while in the control group using the traditional treatment regimen it ended on the 9th day.

Evaluation of the Efficacy and Safety of Silver Nanoparticles in the Treatment of Non-Neurological and Neurological Distemper in Dogs: A Randomized Clinical Trial.

Canine distemper is caused by canine distemper virus (CDV), a multisystemic infectious disease with a high morbidity and mortality rate in dogs. Nanotechnology represents a development opportunity for new molecules with antiviral effects that may become effective treatments in veterinary medicine. This study evaluated the efficacy and safety of silver nanoparticles (AgNPs) in 207 CDV, naturally infected, mixed-breed dogs exhibiting clinical signs of the non-neurological and neurological phases of the disease. Group 1a included 52 dogs (experimental group) diagnosed with non-neurologic distemper treated with 3% oral and nasal AgNPs in addition to supportive therapy. Group 1b included 46 dogs (control group) diagnosed with non-neurological distemper treated with supportive therapy only. Group 2a included 58 dogs with clinical signs of neurological distemper treated with 3% oral and nasal AgNPs in addition to supportive therapy. Group 2b included 51 dogs (control group) diagnosed with clinical signs of neurological distemper treated with supportive therapy only. Efficacy was measured by the difference in survival rates: in Group 1a, the survival rate was 44/52 (84.6%), versus 7/46 in Group 1b (15.2%), while both showed clinical signs of non-neurological distemper. The survival rate of dogs with clinical signs of neurological distemper in Group 2a (38/58; 65.6%) was significantly higher than those in Control Group 2b (0/51; 0%). No adverse reactions were detected in experimental groups treated with AgNPs. AgNPs significantly improved survival in dogs with clinical signs of neurological and non-neurological distemper. The use of AgNPs in the treatment of neurological distemper led to a drastic increase in the proportion of dogs recovered without sequels compared to dogs treated without AgNPs. The evidence demonstrates that AgNP therapy can be considered as a targeted treatment in dogs severely affected by canine distemper virus.

A network theoretic method for the basic reproductive number for infectious diseases

Abstract When an outbreak of an infectious disease occurs, whether it is COVID‐19 in humans, pine wilt in trees or canine distemper in dogs, quick and decisive actions need to be taken to contain it. One tool that can help both the scientific and applied management communities understand the infection risk during an outbreak is the basic reproductive number, . In this paper, we use a network method to calculate and analyse the basic reproductive number, specifically flow network theory. We convert traditional compartmental models to flow networks and then apply the fundamental Max‐Flow Min‐Cut Theorem to calculate the basic reproductive number. We show that this method is equivalent to the traditional next generation matrix method for the calculation of , and thus a valid alternative. Then we provide step‐by‐step instructions and illustrate how to apply this method to epidemic models. The current methods available for calculating are complicated, requiring mathematical training. This can act as a barrier to understanding and cause delays in a real‐time response. Our new approach drastically reduces the mathematical complexity of the calculation and is far more accessible to the broader scientific community. It also allows for novel insights and can be applied to models/situations where traditional methods fail.

Clinical and hematological evaluation in dogs with myoclonus derived from canine distemper supplemented with vitamin D3

In dogs, the synthesis of vitamin D in the skin is considered inefficient, making dietary supplementation the main source of this vitamin for these animals. In humans, there are established values for 25-hydroxyvitamin D (25(OH)D) deficiency, insufficiency and sufficiency levels, however in dogs, the serum concentrations of these values are not well established. The purposes of this study were to evaluate the 25(OH)D serum levels in dogs carrying myoclonus as sequelae of distemper, to evaluate the response to vitamin D levels on oral supplementation, to evaluate PTH, calcium, phosphorus, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood count and leukogram levels, in addition to conduct clinical observations of myoclonus. Venous blood samples were collected from nine dogs carrying myoclonus derived from distemper, however with no other clinical or laboratorial change, of varied breeds and same age group (1 - 8 years old). Screening laboratory tests were performed to attest to the health of the animals in a 30-day period and the collections were divided into three periods: days 0, 15 and 30. After the initiation of treatment, the animals underwent physical and laboratorial evaluations every 15 days, for 90 days, completing a total of 120 days. The dose used for oral supplementation of vitamin D3 was 1000IU/kg administered every day, once a day, during the entire experimental period. For clinical evaluation, parameters of anatomical distribution, speed and rhythm, and distribution of myoclonic changes over time were observed. The laboratory results were subjected to analysis of variance and, when significant (P<0.05), submitted to regression analysis. Descriptive statistics were used to analyze clinical results. There was a significant difference in blood concentrations of 25(OH)D, PTH, calcium and phosphorus, however there was no significant effect of vitamin D on the other parameters evaluated. It was possible to conclude that the dose of vitamin D3 used was sufficient to increase 25(OH)D serum levels in the blood, to levels of sufficiency, having influence on PTH, phosphorus and calcium levels, not changing the other hematological and clinical parameters evaluated. However, the dose and duration of the treatment used did not change the myoclonus derived from distemper in dogs.

Evaluation of infection with N protein-specific Immunoglobulin M and G in naturally occurring distemper in dogs

In dogs, canine distemper has a worldwide distribution with high morbidity/mortality, despite the widespread usage of vaccines and has no specific treatment. In susceptible animals with the canine distemper virus, respiratory, gastrointestinal and nervous system disorders, immunosuppression and cutaneous lesions can also be seen. Especially puppies and unvaccinated dogs are prone to get the viral infection. IgM and IgG antibodies constitute the major component of the natural antibodies produced during the primary and secondary antibody response that have long been recognised to inhibit viral infections. In the present study, the presence of the viral N protein-specific IgM and IgG was investigated by indirect ELISA in naturally infected dogs. Moreover, the rate of outbreaks in naturally infected dogs was shown by the detection of new and re-infections. In the Western Mediterranean region, blood serum samples were collected from 50 unvaccinated dogs for the mentioned infection between 2015 and 2017. At 0–12 months, in the dogs with clinical symptoms, the indirect ELISA detected 4% acute, 54% early convalescent, 40% late convalescent and 2% no infections phases. The clinical manifestations were studied in four main groups follow as: respiratory, gastrointestinal, nervous and cutaneous symptoms. The evaluation showed that the canine distemper virus N protein-specific antibodies detection by the indirect ELISA is quick and safe in naturally infected dogs. In conclusion, the method is very useful for the pre-diagnosis of the disease when evaluated together with the clinical symptoms. It helps to distinguish acute and convalescent (early/late) phases. Distinguishing these phases of infection is important for monitoring the spread of the outbreaks and identifying the risk of severe forms of canine distemper.

Systemic Candidosis in Dogs Associated with Canine Distemper Virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidiasis associated to canine distemper in dogs, highlighting the main epidemiological, clinical and pathological aspects.Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were strongly positive for the anti-Candida albicans polyclonal antibody in the immunohistochemistry. All 3 dogs were concomitantly infected by the canine distemper virus, and 2 of the animals also presented other comorbidities.Discussion: The diagnosis of systemic candidiasis was made based on the morphotintorial characteristics of the agent and confirmed by immunohistochemistry. The yeasts of the genus Candida live as commensals, but when there is an imbalance of the normal microbiota or the immune system of the host is impaired, the yeasts can convert themselves into opportunistic pathogenic microorganisms. The concomitant infection with the canine distemper virus may have favored the infection and proliferation of the agent in the tissues, as the virus causes immunosuppression. The extensive vascular lesions associated to fungi in the vascular lumen and wall observed in the affected organs suggest hematogenous dissemination causing acute infarctions. The clinical signs of systemic candidosis are extremely variable and reflect the affected organs and hardly ever the infections are considered in the differential diagnoses. The dogs presented neurological clinical signs compatible with canine distemper, however the other clinical alterations were unspecific and insufficient to correlate with a secondary infection and none of the cases diagnosed in the necroscopic examination suggested systemic candidiasis during the clinical investigation. The histochemical methods used in this study contributed effectively for the prior diagnosis of candidosis, making evident the variable morphological characteristics of the fungi. It is concluded that the systemic candidosis in dogs with canine distemper affects young and adult animals with predominantly neurological and unspecific clinical signs resulting from necrossupurative and vascular lesions in several organs, including the articular involvement little described in dogs and testicular lesions only reported in humans.

Linking pollution and infectious disease

Thousands of seals died along the coasts of the heavily polluted Baltic Sea in the late 1980s. Scientists traced the deaths to a virus similar to the one that causes distemper in dogs. Last year, the same virus struck hundreds of seals in Maine. In both instances, researchers believe that persistent organic pollutants, such as polychlorinated biphenyls (PCBs), dioxins, and furans, played an indirect role in the seals’ demise. The seals are one example of a phenomenon of increasing importance to toxicologists: the interplay between exposure to environmental contaminants and infectious disease. More than two decades ago, researchers reported that exposure to low levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most toxic of the dioxins, decreases resistance to an influenza virus in mice (Fundam. Appl. Toxicol. 1996, DOI: 10.1006/faat.1996.0004). Scientists have since shown that exposure to other chemicals, including perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA), mercury, and...

Ocorrência de parvovirose e cinomose em cães no Planalto Catarinense

This study determined the occurrence of parvovirus and distemper in dogs treated at the Veterinary Clinics Hospital (HCV) of Universidade Estadual de Santa Catarina (UDESC). For such, 3198 medical records filed in the HCV, from February 2013 to July 2015, were reviewed. From the records, 145 (4.53%) dogs were diagnosed with parvovirus (76 [2.38%]) or distemper (69 [2.15%]). The most common clinical signs of parvovirus were gastrointestinal signs (76 [100%]), having hematochezia as the most commonly observed (74 [97.37%]); for distemper, neurological signs (48 [69.56%]) were most frequently described and myoclonus had the highest occurrence (31 [64.58%]). Regarding complete blood count, the most observed variation in parvovirus was leukopenia by neutropenia and lymphopenia (20 [71.43%]). Distemper showed lymphopenia (28 [59.57%]). It was also observed that the occurrence of distemper and parvovirus were higher according to some selection criteria: breed, age, outdoor access, having contact with other animals and vaccination status. Young non-vaccinated dogs with outdoor access had the highest occurrence ratio. This study showed that viral infectious diseases presented low occurrence, and parvovirus was the most reported agent.

Update on canine distemper virus (CDV) strains of Arctic-like lineage detected in dogs in Italy.

Canine distemper virus (CDV) is the etiologic agent of distemper in dogs. It exhibits an elevated potential of crossing species barriers, infecting a wide range of wild and domestic carnivores. Of its encoding genes, hemagglutinin (H) shows high heterogeneity, and it was used to determine the relationship between CDV strains due to its variability and key role in determining cell tropism, host shift, and in eliciting a protective immune response. This study analysed the full-length H gene sequence of Arctic-like CDV strains collected from dogs in Italy during a period in which an increased activity of CDV diffusion was observed. The common amino acid changes and features of Arctic-like CDV strains collected from 2011 to 2016 in Europe were described, providing an updated analysis of the genomic features. A comparison with CDV vaccine strains was carried out to evaluate the increased genomic difference with CDV Arctic-like field strains. This study provides a complete and updated analysis of the current spreading strains of Arctic-like lineage and the main amino acid variations in the hemagglutinin gene sequence circulating in Italy. Moreover, it provides novel information regarding the evolution of the most recent CDV Arctic-like lineage strains collected in Europe.

Infectious Progression of Canine Distemper Virus from Circulating Cerebrospinal Fluid into the Central Nervous System

In the current study, we generated recombinant chimeric canine distemper viruses (CDVs) by replacing the hemagglutinin (H) and/or phosphoprotein (P) gene in an avirulent strain expressing enhanced green fluorescent protein (EGFP) with those of a mouse-adapted neurovirulent strain. An in vitro experimental infection indicated that the chimeric CDVs possessing the H gene derived from the mouse-adapted CDV acquired infectivity for neural cells. These cells lack the CDV receptors that have been identified to date (SLAM and nectin-4), indicating that the H protein defines infectivity in various cell lines. The recombinant viruses were administered intracerebrally to 1-week-old mice. Fatal neurological signs of disease were observed only with a recombinant CDV that possessed both the H and P genes of the mouse-adapted strain, similar to the parental mouse-adapted strain, suggesting that both genes are important to drive virulence of CDV in mice. Using this recombinant CDV, we traced the intracerebral propagation of CDV by detecting EGFP. Widespread infection was observed in the cerebral hemispheres and brainstems of the infected mice. In addition, EGFP fluorescence in the brain slices demonstrated a sequential infectious progression in the central nervous system: CDV primarily infected the neuroependymal cells lining the ventricular wall and the neurons of the hippocampus and cortex adjacent to the ventricle, and it then progressed to an extensive infection of the brain surface, followed by the parenchyma and cortex. In the hippocampal formation, CDV spread in a unidirectional retrograde pattern along neuronal processes in the hippocampal formation from the CA1 region to the CA3 region and the dentate gyrus. Our mouse model demonstrated that the main target cells of CDV are neurons in the acute phase and that the virus spreads via neuronal transmission pathways in the hippocampal formation. CDV is the etiological agent of distemper in dogs and other carnivores, and in many respects, the pathogenesis of CDV infection in animals resembles that of measles virus infection in humans. We successfully generated a recombinant CDV containing the H and P genes from a mouse-adapted neurovirulent strain and expressing EGFP. The recombinant CDV exhibited severe neurovirulence with high mortality, comparable to the parental mouse-adapted strain. The mouse-infectious model could become a useful tool for analyzing CDV infection of the central nervous system subsequent to passing through the blood-cerebrospinal fluid barrier and infectious progression in the target cells in acute disease.

Silver nanoparticles composition for treatment of distemper in dogs

Canine distemper is caused by the canine distemper virus (CDV), a paramyxovirus of the genus Morbillivirus, with a worldwide distribution. Canine distemper is a multisystem disease, potentially fatal. Affected organisms may develop clinical or subclinical infections, among them respiratory, gastrointestinal and of the nervous system. In the present work, we use a veterinary pharmaceutical composition of silver nanoparticles (AgNPs) for the treatment of distemper in dogs with neurological and non-neurological symptoms. AgNPs treatment of animals with non-neurological distemper provides a very high rate of recovery without sequelae. However, in dogs with neurological symptoms, AgNPs treatment was not able to reverse the disease.

Clinical, pathological, and immunohistochemistry characterization of toxoplasmosis in dogs with distemper in the semiarid region of Paraíba, Brazil

<p>The objective of this study was to describe the clinical, pathological, and immunohistochemistry characteristics of five cases of toxoplasmosis, an infection often associated with distemper in dogs. From January 2000 to December 2012, a retrospective study was performed analyzed dogs with distemper in the semiarid region of Paraíba. We evaluated this sample to focus on individuals who presented with concomitant structures in protozoa characteristics, and performed immunohistochemistry (IHC) tests using polyclonal anti-<em>Toxoplasma gondii </em>antibody. In all cases, the clinical signs were similar including digestive changes, as well as respiratory, neurological, and ocular lesions, suggesting an infection of canine distemper virus. The diagnosis of distemper was confirmed on histopathological analysis depending on the presence of intranuclear and intracytoplasmic eosinophilic inclusion bodies in different tissues. Histopathological examination also revealed the characteristic presence of parasitic cysts <em>T. gondii </em>in the brain in four cases, and in the lung in one case. The brain cysts were associated with multifocal areas of malacia and lung there was alveolar septa thickening due to infiltration of macrophages, lymphocytes, and plasma cells, with moderate proliferation of type II pneumocytes and coalescing multifocal areas of necrosis. These cysts are characterized by round and strongly basophilic structures, measuring approximately 5 to 70 ?m, delimited by thin wall, stained by hematoxylin and eosin, and immunomarked as brown by immunohistochemistry (IHC) using the chromogen DAB. The diagnosis of toxoplasmosis associated with infection by canine distemper virus in the five case studied was based on microscopic findings and confirmed by immunohistochemistry. Toxoplasmosis should be included in the differential diagnosis of dogs with severe progressive systemic signs, especially when respiratory and neurological involvement is suspected.</p>

Clinical, pathological, and immunohistochemistry characterization of toxoplasmosis in dogs with distemper in the semiarid region of Paraíba, Brazil

The objective of this study was to describe the clinical, pathological, and immunohistochemistry characteristics of five cases of toxoplasmosis, an infection often associated with distemper in dogs. From January 2000 to December 2012, a retrospective study was performed analyzed dogs with distemper in the semiarid region of Paraíba. We evaluated this sample to focus on individuals who presented with concomitant structures in protozoa characteristics, and performed immunohistochemistry (IHC) tests using polyclonal anti-Toxoplasma gondii antibody. In all cases, the clinical signs were similar including digestive changes, as well as respiratory, neurological, and ocular lesions, suggesting an infection of canine distemper virus. The diagnosis of distemper was confirmed on histopathological analysis depending on the presence of intranuclear and intracytoplasmic eosinophilic inclusion bodies in different tissues. Histopathological examination also revealed the characteristic presence of parasitic cysts T. gondii in the brain in four cases, and in the lung in one case. The brain cysts were associated with multifocal areas of malacia and lung there was alveolar septa thickening due to infiltration of macrophages, lymphocytes, and plasma cells, with moderate proliferation of type II pneumocytes and coalescing multifocal areas of necrosis. These cysts are characterized by round and strongly basophilic structures, measuring approximately 5 to 70 ?m, delimited by thin wall, stained by hematoxylin and eosin, and immunomarked as brown by immunohistochemistry (IHC) using the chromogen DAB. The diagnosis of toxoplasmosis associated with infection by canine distemper virus in the five case studied was based on microscopic findings and confirmed by immunohistochemistry. Toxoplasmosis should be included in the differential diagnosis of dogs with severe progressive systemic signs, especially when respiratory and neurological involvement is suspected.

Systemic acanthamoebiasis associated with canine distemper in dogs in the semiarid region of Paraíba, Brazil

Infections by free-living amoebae can cause systemic disease in animals and humans. We describe the epidemiological, clinical and pathological aspects of disseminated acanthamoebiasis associated with canine distemper in three dogs of the semiarid region of Paraíba, Northeastern Brazil. Affected dogs developed progressive neurological and respiratory signs that progressed to death within in two to 20 days. Gross lesions were irregular and with yellow-reddish nodules randomly distributed in the lungs, heart, kidneys, spleen, lymph nodes, adrenals, and intestine. One dog had foci of malacia in the parietal cortex and another one in nucleus of brain basis. Histologically, pyogranulomas with areas of necrosis and hemorrhage in all organs affected were observed, associated with myriads of intralesional amoebic trophozoites. All three cases were concomitant canine distemper, that possibly triggered immunosuppression in the dogs. The diagnosis was performed through microscopic findings of infection by free-living amoebae and confirmed Acanthamoeba sp. by immunohistochemistry

Virological and serological findings in dogs with naturally occurring distemper

Canine distemper virus (CDV) is the cause of a severe and highly contagious disease in dogs. The unpredictable and variable course of CDV-related disease may hamper correct diagnosis of infection and makes it crucial the collection of samples suitable for laboratory confirmation. In the present study we were able to follow the disease in two dogs infected naturally, collecting different biological matrices during the entire period of infection. By real time RT-PCR, viral RNA was detected and quantified, suggesting that urine and rectal swabs would be useful for ante-mortem diagnosis of distemper in dogs, regardless of the clinical stage and form of the illness.

Antimicrobial activity, toxicity and selectivity index of two biflavonoids and a flavone isolated from Podocarpus henkelii (Podocarpaceae) leaves.

BackgroundDifferent parts of Podocarpus henkelii have been used in many cultures around the world to treat ailments such as cholera, stomach diseases, rheumatism, cancer, canine distemper in dogs and gall sickness in cattle. The aim of this study was to evaluate the biological activity and toxicity of isolated compounds from Podocarpus henkelii after an earlier study indicated a promising activity in crude extracts against viral pathogens of veterinary importance.MethodsThe antibacterial and antifungal activity of two biflavonoids 7, 4’, 7”, 4”’-tetramethoxy amentoflavone (TMA), isoginkgetin (IGG) and podocarpus flavone–A (PFA) isolated from the leaves of Podocarpus henkelii were determined using a serial microplate dilution method with tetrazolium violet as growth indicator. The cytotoxicity of compounds TMA and IGG were determined on different cell types using a tetrazolium-based colorimetric cellular assay (MTT). The Ames test was used to determine their mutagenic activities.ResultsTMA had reasonable antifungal activity against Aspergillus fumigatus (MIC = 30 μg/ml). IGG had a wide spectrum of activity against four bacterial and two fungal pathogens with much higher selectivity index values obtained for A. fumigatus and Cryptococcus neoformans (SI > 30). PFA had a broad spectrum of activity against Enterococcus faecalis and Pseudomonas aeruginosa (SI > 15) and less activity against the two fungal pathogens. In both the cytotoxicity assays and Ames mutagenicity test using Salmonella typhimurium strains TA98 and TA100, TMA and IGG had no deleterious effect on the different cell types and did not induce mutations in the Ames test.ConclusionAlthough the antimicrobial activities of the isolated compounds were not that exciting, the compounds had no cytotoxic activity at the highest concentration (1000 μg/ml) tested against all three cell lines. IGG was the most active against E. coli, S. aureus, A. fumigatus and C. neoformans, exhibiting both antibacterial and antifungal activity with good selectivity index values. PFA had a broad spectrum of activity against E. faecalis and P. aeruginosa. The two compounds isolated had low toxicity and no genotoxic activity in the Ames test.

Whole genome sequence analysis of the arctic-lineage strain responsible for distemper in Italian wolves and dogs through a fast and robust next generation sequencing protocol

Dynamic surveillance and characterization of canine distemper virus (CDV) circulating strains are essential against possible vaccine breakthroughs events. This study describes the setup of a fast and robust next-generation sequencing (NGS) Ion PGM™ protocol that was used to obtain the complete genome sequence of a CDV isolate (CDV2784/2013). CDV2784/2013 is the prototype of CDV strains responsible for severe clinical distemper in dogs and wolves in Italy during 2013. CDV2784/2013 was isolated on cell culture and total RNA was used for NGS sample preparation. A total of 112.3Mb of reads were assembled de novo using MIRA version 4.0rc4, which yielded a total number of 403 contigs with 12.1% coverage. The whole genome (15,690bp) was recovered successfully and compared to those of existing CDV whole genomes. CDV2784/2013 was shown to have 92% nt identity with the Onderstepoort vaccine strain. This study describes for the first time a fast and robust Ion PGM™ platform-based whole genome amplification protocol for non-segmented negative stranded RNA viruses starting from total cell-purified RNA. Additionally, this is the first study reporting the whole genome analysis of an Arctic lineage strain that is known to circulate widely in Europe, Asia and USA.

Clinical syndromes of nervous distemper in dogs initially presented without conventional evidences of CDV infection

Apesar da pratica de vacinacao, o virus da cinomose canina (CDV) ainda e um importante agente infeccioso que leva a doenca neurologica na populacao canina em todo o mundo. Infelizmente, o diagnostico clinico da encefalomielite pela cinomose e dificil nos casos apresentados sem evidencias clinicas convencionais de infeccao pelo CDV, tais como sinais sistemicos e mioclonia. Portanto, o objetivo deste estudo foi avaliar os deficits neurologicos e as sindromes clinicas da apresentacao neurologica da cinomose em caes portadores de doenca neurologica apresentados na ausencia de evidencias convencionais de infeccao pelo CDV. Foram prospectivamente acompanhados caes que apresentaram doenca do sistema nervoso central, onde a abordagem diagnostica ante mortem excluiu outras afeccoes que nao cinomose (etiologia toxica, traumatica, degenerativa, neoplasica, etc), sendo incluidos neste estudo os animais que vieram a obito (morte natural ou eutanasia, apesar do tratamento medico) e foram necropsiados. Dez de 35 caes avaliados foram diagnosticados no post-mortem com encefalomielite pelo CDV pela deteccao de RNA viral pelo RT-PCR no tecido nervoso e observacao de lesoes neuroparenquimatosas compativeis com cinomose. De acordo com os sinais neurologicos, a historia clinica e a idade de apresentacao, os caes foram agrupados em tres sindromes clinicas da forma neurologica da cinomose: i) encefalite pela cinomose canina em caes imaturos (CDEID) ( n = 3); ii) encefalomielite multifocal pela cinomose em caes adultos (MDEMD) ( n = 6); e iii) sindrome similar a encefalite do cao velho (ODE-like) ( n = 1). Os respectivos deficits neurologicos esperados para cada uma das sindromes aqui apresentadas foram discutidos.

Recombinant Canine Distemper Virus Strain Snyder Hill Expressing Green or Red Fluorescent Proteins Causes Meningoencephalitis in the Ferret

The propensity of canine distemper virus (CDV) to spread to the central nervous system is one of the primary features of distemper. Therefore, we developed a reverse genetics system based on the neurovirulent Snyder Hill (SH) strain of CDV (CDV(SH)) and show that this virus rapidly circumvents the blood-brain and blood-cerebrospinal fluid (CSF) barriers to spread into the subarachnoid space to induce dramatic viral meningoencephalitis. The use of recombinant CDV(SH) (rCDV(SH)) expressing enhanced green fluorescent protein (EGFP) or red fluorescent protein (dTomato) facilitated the sensitive pathological assessment of routes of virus spread in vivo. Infection of ferrets with these viruses led to the full spectrum of clinical signs typically associated with distemper in dogs during a rapid, fatal disease course of approximately 2 weeks. Comparison with the ferret-adapted CDV(5804P) and the prototypic wild-type CDV(R252) showed that hematogenous infection of the choroid plexus is not a significant route of virus spread into the CSF. Instead, viral spread into the subarachnoid space in rCDV(SH)-infected animals was triggered by infection of vascular endothelial cells and the hematogenous spread of virus-infected leukocytes from meningeal blood vessels into the subarachnoid space. This resulted in widespread infection of cells of the pia and arachnoid mater of the leptomeninges over large areas of the cerebral hemispheres. The ability to sensitively assess the in vivo spread of a neurovirulent strain of CDV provides a novel model system to study the mechanisms of virus spread into the CSF and the pathogenesis of acute viral meningitis.