CYTOPATHOGENICITY OF INFLUENZA VIRUS (H4 N3)

Monolayer tissue cultures of chicken embryo fibroblast ( CEF ) cells infected with avian influenza virus isolate were examined by the hematoxylie and eosin (H&E) staining and indirect immunoperoxidase test for studying the cytopathogenic effect of the virus. Cytopathological changes which occurred in the uncleus of infected cells included nuclear and nucleolar hypertropy, chromatin margination and intranuclear inclusions. The most striking cytoplasmic change were the presence of perinuclear. eosinophilic inclusions at 22-36 hours post inoculation ( p. i.). Vacuolization, and granulation were also observed. Indirect immunoperoxidase ( IP ) test demonstrated the localization of influenza virus antigens in infected cells. A positive peroxidase reaction observed in the nucleus and cytoplasm were similar to those shown hematoxyline and eosin staining.

Detection and Molecular Characterization of Two Gammaherpesviruses from Pantesco Breed Donkeys during an Outbreak of Mild Respiratory Disease

Equid and asinine gammaherpesviruses (GHVs; genus Percavirus) are members of the Herpesviridae family. Though GHVs have been reported in horse populations, less studies are available on gammaherpesviral infections in donkeys. This study reports the co-infection with two GHVs in Pantesco breed donkeys, an endangered Italian donkey breed. Samples (n = 124) were collected on a breeding farm in Southern Italy from 40 donkeys, some of which were healthy or presented erosive tongue lesions and/or mild respiratory signs. Samples were analysed by using a set of nested PCRs targeting the DNA polymerase, glycoprotein B, and DNA-packaging protein genes, and sequence and phylogenetic analyses were performed. Twenty-nine donkeys (72.5%) tested positive, and the presence of Equid gammaherpesvirus 7 and asinine herpesvirus 5 was evidenced. In 11 animals, we found evidence for co-infection with viruses from the two species. Virions with herpesvirus-like morphology were observed by electron microscopic examination, and viruses were successfully isolated in RK-13-KY cell monolayers. The histological evaluation of tongue lesions revealed moderate lympho-granulocytic infiltrates and rare eosinophilic inclusions. The detection of GHVs in this endangered asinine breed suggests the need long-life monitoring within conservation programs and reinforces the need for further investigations of GHV’s pathogenetic role in asinine species.

Characterization of Urothelial Inclusions in Male Wistar Han Rats Treated Orally With the Novel α2A-Adrenoceptor Agonist Tasipimidine

Intracellular inclusions were observed in urinary bladder epithelium of male Wistar rats, following oral treatment with high doses of the α2A-adrenoceptor agonist tasipimidine for 28 days. No cell death or inflammation was associated with the brightly eosinophilic inclusions. Electron microscopy (EM) studies showed that the inclusions represented intact or fragmented red blood cells (RBC) resulting from erythrophagocytosis, further supported by the presence of iron in urothelial cells. In addition, scattered iron-positive macrophages were observed in the submucosa and muscle layer, indicating microvascular leakage, as no major hemorrhage was evident. Despite the presence of inclusions, the urothelium showed normal uroplakin III distribution, normal cell turnover, and an absence of α-2u-globulin. It is, therefore, concluded that the inclusions were not associated with urothelial damage or increased renewal of the epithelium. This finding shows also that urothelial cells have the capability to phagocytize and break down RBCs originating from submucosal microvascular leakage. Similar changes were not observed in tasipimidine-treated beagle dogs (28 days), suggesting these findings were rat specific. The leakage of RBCs into the urothelium is suggested to be a consequence of exaggerated pharmacology leading to vasoconstriction of submucosal blood vessels in combination with transient increased bladder distension and pressure.

The clinical, histologic, and genotypic spectrum of SEPN1-related myopathy

ObjectiveTo clarify the prevalence, long-term natural history, and severity determinants of SEPN1-related myopathy (SEPN1-RM), we analyzed a large international case series.MethodsRetrospective clinical, histologic, and genetic analysis of 132 pediatric and adult patients (2–58 years) followed up for several decades.ResultsThe clinical phenotype was marked by severe axial muscle weakness, spinal rigidity, and scoliosis (86.1%, from 8.9 ± 4 years), with relatively preserved limb strength and previously unreported ophthalmoparesis in severe cases. All patients developed respiratory failure (from 10.1±6 years), 81.7% requiring ventilation while ambulant. Histopathologically, 79 muscle biopsies showed large variability, partly determined by site of biopsy and age. Multi-minicores were the most common lesion (59.5%), often associated with mild dystrophic features and occasionally with eosinophilic inclusions. Identification of 65 SEPN1 mutations, including 32 novel ones and the first pathogenic copy number variation, unveiled exon 1 as the main mutational hotspot and revealed the first genotype–phenotype correlations, bi-allelic null mutations being significantly associated with disease severity (p = 0.017). SEPN1-RM was more severe and progressive than previously thought, leading to loss of ambulation in 10% of cases, systematic functional decline from the end of the third decade, and reduced lifespan even in mild cases. The main prognosis determinants were scoliosis/respiratory management, SEPN1 mutations, and body mass abnormalities, which correlated with disease severity. We propose a set of severity criteria, provide quantitative data for outcome identification, and establish a need for age stratification.ConclusionOur results inform clinical practice, improving diagnosis and management, and represent a major breakthrough for clinical trial readiness in this not so rare disease.

Atypical presentation of genital molluscum contagiosum mimicking genital warts

A man in his 20s with a history of unprotected sex with a commercial sex worker three months before presented with a solitary slowly-growing, well-defined, non-tender, oval, verrucous lesion on the mucosal aspect of his prepuce. Dermoscopy revealed a polylobular, pinkish-white, amorphous structure in the centre with a surrounding crown of vessels that did not cross the centre of the lobules, and histopathology showed keratinocytes containing intracytoplasmic faintly granular eosinophilic inclusions and peripherally-pushed nuclei. A diagnosis of agminated genital molluscum contagiosum was reached on the basis of the above findings. Molluscum contagiosum can present with several atypical morphologies mimicking other dermatoses. Agminated molluscum contagiosum can closely mimic a genital wart especially if it is involving the ano-genital region. Awareness among clinicians regarding the atypical presentations of molluscum contagiosum is imperative for early diagnosis and management of this infectious condition.

Rabies-like neuronal inclusions in a dog with meningoencephalomyelitis and neuritis of unknown origin

A four-year-old intact female pointer dog from a hunting plantation in South Georgia, USA, developed acute neurological signs. The referring veterinarian suspected rabies or pseudorabies; the dog was euthanased and submitted to the Tifton Veterinary Diagnostic and Investigational Laboratory for necropsy. No gross anatomical changes were present. Portions of brain were submitted for rabies virus fluorescent antibody test with a negative result. Histopathology revealed a marked lymphoplasmacytic meningoencephalitis with numerous intracytoplasmic eosinophilic inclusions within neurons, still raising concerns for rabies meningoencephalitis. Rabies immunohistochemistry was then performed on the brain, yielding a negative result. Brain samples were also negative for canine distemper virus and herpesvirus (pan-herpesvirus primers) PCR. Electron microscopy did not reveal viral particles within the inclusions. Similar intraneuronal pseudoinclusions have been previously reported in non-rabid dogs and cats. Such inclusions are a diagnostic challenge, especially in animals with central neurological signs and/or meningoencephalitis.

Microscopic changes in the colon of puppies at experimental infection with parvovirus isolator cultivated in heterologous cell culture

The article presents the results of the study of histological changes in the cecum, and colon of dogs for experimental infection with parvovirus enteritis. The histological examination of the small intestine, selected from corpses (n = 5) of puppies, dental labradorus with unborn, was infected with field isolator of parvovirus cultured on heterologous cell cultures (kidney kidney hamster (BHK-21), rabbit kidney (RK-13) and the renal mumps (SPEV). The presence of parvovirus, without any other association in experimental animals, is confirmed by the ELISA method and the solid-phase ELISA system. The purpose of this work was to study and characterize the microscopic changes in the colon for experimental contamination of dogs by parvovirus enteritis. The work was carried out at the Faculty of Veterinary Medicine of Zhytomyr National Agroecological University (ZNAEU). Anatomy of the animals was carried out in the special laboratory of the Department of Anatomy and Histology. The material of the study was pathological material taken during the pathoanatomical dissect of the puppies (n = 5), after experimental infection and euthanasia. The section of the cecum and the сolon, were investigated. The main method used in the work was a histological study, and a description of the microstructural changes in the tissues of the organs. Histological studies of the bladder and intestine showed that microscopic changes in their walls were segmental. Some parts had well preserved the spots. Only moderate uneven swelling of the submucosal base, an increase in its blood vessels, and edema and partial lysis of collagen fibers are registered. In other areas, destruction was recorded. In some cases, only the upper part of the crypt disappeared. The cript stroma was disorganized and suppressed, while in the nuclei of a sufficiently large number of epithelial cells in the upper crypts, eosinophilic inclusions were recorded. In the large intestine, puppies for experimental infection with parvovirus isolate, cultured in the heterologous culture of the lesions, are localized in the cecum and the colon and have a segmental nature: in some areas the crypts is well preserved, while in others the destruction of the crypts is recorded. Local reaction of the system of specific immunity is characterized by hypertrophy of isolated and congested lymphoid nodes. There are small cells of lymphocyte necrosis. In the nuclei of the epithelial cytoplasm, as well as in the nuclei of the lymphocytes, eosinophilic inclusions of the corpuscles are detected.

Clinical and neuropathological features of ALS/FTD with TIA1 mutations

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) represent a disease continuum with common genetic causes and molecular pathology. We recently identified mutations in the T-cell restricted intracellular antigen-1 (TIA1) protein as a cause of ALS +/− FTD. TIA1 is an RNA-binding protein containing a low complexity domain (LCD) that promotes the assembly of membrane-less organelles, such as stress granules (SG). Whole exome sequencing of two family members with fALS/FTD revealed a novel missense mutation in the TIA1 LCD (P362L). Subsequent screening identified five more TIA1 mutations in six additional ALS patients, but none in controls. All mutation carriers presented with weakness, behavioral abnormalities or language impairments and had a final diagnosis of ALS +/− FTD. Autopsy on five TIA1 mutation carriers showed widespread neurodegeneration with TDP-43 pathology. Round eosinophilic inclusions in lower motor neurons were a consistent feature. Cellular assays revealed abnormal SG dynamics in the presence of TIA1 mutations. In summary, missense mutations in the LCD of TIA1 are a newly recognized cause of ALS/FTD with TDP-43 pathology and strengthen the role of RNA metabolism in the pathogenesis in this disease.

Confirmation and Sequence analysis of N gene of PPRV in South Xinjiang, China

ABSTRACT In China, Peste des petits ruminants (PPR) was officially first reported in 2007. From 2010 until the outbreak of 2013, PPRV infection was not reported. In November 2013, PPRV re-emerged in Xinjiang and rapidly spread to 22 P/A/M (provinces, autonomous regions and municipalities) of China. In the study, suspected PPRV-infected sheep in a breeding farm of South Xinjiang in 2014 were diagnosed and the characteristics of complete sequence of N protein gene of PPRV was analyzed. The sheep showed PPRV-infected signs, such as fever, orinasal secretions increase, dyspnea and diarrhea, with 60% of morbidity and 21.1% of fatality rate. The macroscopic lesions after autopsy and histopathological changes were observed under light microscope including stomatitis, broncho-interstitial pneumonia, catarrhal hemorrhagic enteritis and intracytoplasmic eosinophilic inclusions in multinucleated giantcell in lung. The formalin-fixed mixed tissues samples were positive by nucleic acid extraction and RT-PCR detection. The nucleotide of N protein gene of China/XJNJ/2014 strain was extremely high homology with the China/XJYL/2013 strain, and the highest with PRADESH_95 strain from India in exotic strains. Phylogenetic analysis based on complete sequence of N protein gene of PPRV showed that the China/XJNJ/2014 strain, other strain of 2013-2014 in this study and Tibetan strains all belonged to lineage Ⅳ, but the PPRV strains of 2013-2014 in this study and Tibetan strains were in different sub-branches.

An outbreak of sheep pox in an organized farm of Tamil Nadu, India

Outbreak of sheep pox was noticed in flocks of Khilakaraisal in an organized government farm of Tirunelveli district during 2013. Mortality and morbidity rates were 4.17% and 80.00%, respectively. Both the sexes of sheep between 6 months to 2 years old were affected. DNA extracted from the scab specimens (n=5) were amplified using Capri pox specific p32 gene primers and yielded 192 bp product. In order to differentiate between sheep pox and goat pox PCR – RFLP was carried out. Digestion of PCR product with EcoRI enzyme yielded two fragments with a size of 123 bp and 63 bp which confirmed the presence of sheep pox virus. Histopathological examination of tissues revealed extensive hydropic degeneration of proliferating epidermal cells with presence of large number of intracytoplasmic eosinophilic inclusion bodies in the keratinocytes of epidermal cells in skin. Lung revealed large infiltration of lymphocytes and macrophages with presence of intracytoplasmic eosinophilic inclusions in the macrophages, proliferation of type II pneumocytes were prominent along with bronchiolar epithelial hyperplasia. The outbreak was controlled by effective preventive measures like vaccination of healthy animals, affected animals were treated with a course of antibiotic for 7 days, contaminated areas were disinfected and restricted the movement of animals from unaffected areas to infected areas.