Eosinophilic Intracytoplasmic Inclusion Research Articles

Epidemiologic, clinicopathologic, and diagnostic findings in pet rabbits with myxomatosis caused by the California MSW strain of myxoma virus: 11 cases (2022-2023).

To determine epidemiologic features of naturally occurring myxomatosis in domestic rabbits in California and to characterize clinicopathologic and diagnostic findings. 11 client-owned rabbits, Oryctolagus cuniculus subsp domesticus. A prospective study of pet rabbits with myxomatosis seen at an exotic animal specialty clinic in Santa Cruz county, California, was conducted between January 1, 2022, and December 31, 2023. Rabbits were included in the study if they had bilateral blepharedema and were PCR positive for myxoma virus. All infected rabbits had spent time outdoors. Common clinical signs included bilateral blepharedema (11/11), anogenital edema (10/11), rectal temperature ≥ 39.7 °C (5/9), and sudden death (4/11). Eyelid biopsies from all rabbits (11/11) were positive for myxoma virus by qualitative PCR followed by Sanger sequencing (100% nucleotide identity to strain MSW, also known as California/San Francisco 1950 [Genbank accession KF148065]). Most rabbits had keratinocytes containing eosinophilic intracytoplasmic viral inclusions in biopsies of edematous skin (8/11) and lymphocyte necrosis in the spleen (10/11). Immunohistochemistry identified myxoma virus in samples of skin, heart, lung, ileum, spleen, and lymph node. Clinical signs of myxomatosis caused by the MSW strain of myxoma virus are distinctive but subtle. Cases occur regularly in the Santa Cruz and San Jose regions of California. As infection with this virus is almost 100% fatal and no vaccine is available in the US, owners of domestic rabbits in endemic areas should keep their pets indoors or behind mosquito screens. Myxomatosis is a reportable disease in the US, and the appropriate state or federal agencies should be contacted when outbreaks occur.

Comparative study of non-suppurative meningoencephalitis in cattle in Southern Brazil.

Viral neurologic diseases are common in cattle, although most non-suppurative meningoencephalitis (NSM) remains etiologically unknown. We compared the epidemiological, clinical, and pathological data among 79 cases of rabies, 12 cases of NSM of unknown etiology (NSM-UE), and 8 cases of herpetic meningoencephalitis previously diagnosed in cattle in Southern Brazil. Neurological clinical signs were similar among rabies and NSM-UE and different in cattle with herpetic meningoencephalitis. Only two herpetic meningoencephalitis cases had gross lesions in the central nervous system, characterized by malacia and hemorrhage. Histologically, all three groups had mild to severe multifocal infiltrates of lymphocytes, plasma cells, and macrophages/microglial cells in the Virchow-Robin space, neuropil, and leptomeninges, and gliosis. Other findings included malacia and eosinophilic intracytoplasmic inclusion in rabies, and malacia and intranuclear amphophilic inclusion in herpetic meningoencephalitis. By immunohistochemistry, the predominant inflammatory cells in all cases were T lymphocytes, followed by macrophages/microglial cells, B lymphocytes, and astrocytes. The T lymphocyte count showed statistically significant differences between the diseases. Our results revealed few differences between the groups. Although the etiological agent involved has not been identified in cases of NSM-UE, the characteristics observed in this study showed similarity with viral diseases.

Poster presentationsDP11 Bright, eosinophilic intracytoplasmic inclusion bodies: a rare presentation of acquired epidermodysplasia verruciformis with widespread human papillomavirus infection in a transplant recipient

Poster presentationsDP11 Bright, eosinophilic intracytoplasmic inclusion bodies: a rare presentation of acquired epidermodysplasia verruciformis with widespread human papillomavirus infection in a transplant recipient

Sarcomatoid and Rhabdoid Renal Cell Carcinoma: Clinical, Pathologic, and Molecular Genetic Features.

Renal cell carcinoma (RCC) with sarcomatoid and rhabdoid morphologies has an aggressive biological behavior and a typically poor prognosis. The current 2022 WHO classification of renal tumors does not include them as distinct histologic entities but rather as transformational changes that may arise in a background of various distinct histologic types of RCC. The sarcomatoid component shows malignant spindle cells that may grow as intersecting fascicles, which is reminiscent of pleomorphic undifferentiated sarcoma. The rhabdoid cells are epithelioid cells with eccentrically located vesicular nuclei with prominent nucleoli and large intracytoplasmic eosinophilic inclusions. Studies have shown that RCCs with sarcomatoid and rhabdoid differentiation have distinctive molecular features. Sarcomatoid RCC harbors shared genomic alterations in carcinomatous and rhabdoid components, but also enrichment of specific genomic alterations in the sarcomatoid element, suggesting molecular pathways for development of sarcomatoid growth from a common clonal ancestor. Rhabdoid differentiation also arises through clonal evolution although less is known of specific genomic alterations in rhabdoid cells. Historically, treatment has lacked efficacy, although recently immunotherapy with PD-1/PD-L1/CTLA-4 inhibitors has produced significant clinical responses. Reporting of sarcomatoid and rhabdoid features in renal cell carcinoma is required by the College of American Pathologists and the International Collaboration on Cancer Reporting. This manuscript reviews the clinical, pathologic, and molecular features of sarcomatoid RCC and rhabdoid RCC with emphasis on the morphologic features of these tumors, significance of diagnostic recognition, the molecular mechanisms of tumorigenesis and differentiation along sarcomatoid and rhabdoid lines, and advances in treatment, particularly immunotherapy.

Fatal avian pox in a backyard flock of domestic ducks in Bangladesh

AbstractThis case study describes a fatal avian pox outbreak in a backyard flock of domestic ducks (Anas platyrhynchos domesticus) in Bangladesh. A total of six ducks of varying ages were affected and presented with solid tumour‐like masses on the legs. A 6.5‐month‐old female duck was presented for clinical examination, and a 2 cm3 solid, nodular mass was surgically excised from the left leg. Histopathological examination of the tumour mass revealed marked epidermal hyperplasia with ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies, as well as severe proliferation and necrotising dermatitis, with marked heterophilic and histiocytic infiltration. Electron microscopy revealed numerous dumbbell‐shaped virus particles within the intracytoplasmic inclusion bodies on degenerating keratinocytes, confirming Avipoxvirus infection. This is the first case report of avian pox in ducks in Bangladesh.

Clinicopathological Investigations Among Recurrent Camelpox Outbreaks in Omanis’ Arabian Camels (Camelus dromedarius)

Camelpox remains a widespread viral disease in camelids, with socioeconomic relevance. The present study explored the hematological, biochemical, and histopathological alterations in dromedary-racing camels from the North of Oman infected with camelpox virus diagnosed by real-time PCR. Blood and skin samples were collected from camels with clinical signs and skin lesions (n=4) and from healthy camels (n=3) from 10 different camel herds. The results indicated that the infected camels showed clinical signs, including pyrexia, lacrimation, nasal mucus discharge, affixed and swollen eyelids, emaciation, and pimples on the skin of the head, legs, and abdomen. Hemoglobin, hematocrit, and platelets were significantly greater, with a significant reduction in leukocyte and lymphocyte counts in infected camels than in healthy camels. Infected camels had higher CK and creatinine levels and hepatic-related metabolites, including AST, ALP, AST, GGT, and LDH, than the apparently healthy camels. Histopathological examination of skin scab samples revealed ballooning degeneration of epidermal cells in the presence of typical large eosinophilic intracytoplasmic inclusion bodies and suppurative dermatitis following secondary bacterial infection in all examined infected camels. Camelpox viral DNA was detected using real-time PCR in the blood and skin samples of all infected camels. These findings in dromedary-racing camels associated with a molecular diagnosis of camelpox are described for the first time in the Sultanate of Oman. Therefore, further studies are warranted.

First report of Psittacid Avipoxvirus in Agapornis in Mexico

A postmortem study was performed on two lovebirds (Agapornis fischeri and Agapornis personatus) that had scabs in the periocular region and on the eyelid, as well as serous blepharitis. Microscopically, the eyelids showed ulcers, necrosis and serocellular crusts, severe hyperplasia of keratinocytes with eosinophilic intracytoplasmic inclusion bodies (Bollinger's bodies), bacterial colonies of gram-positive coccoid morphology and PAS-positive septate and 45° branching hyphae. The microbiological study identified the colonies as Staphylococcus spp. and Aspergillus fumigatus, respectively. Using molecular techniques, avian pox clade C was identified on the eyelid. This is the first report in Mexico of a case of avian pox in parrots associated with clade C Avipoxvirus

The Compact Bleb-Eosinophilic Solid and Cystic Renal Cell Carcinoma

The contemporary, indolent, Eosinophilic solid and cystic renal cell carcinoma configures as a predominantly solitary neoplasm delineating solid and cystic pattern of tumour evolution. Neoplastic cells are incorporated with abundant Eosinophilic cytoplasm and intracytoplasmic inclusions or basophilic stippling. Tumour cells display frequent somatic loss of function genetic mutations confined to TSC1 (9q34.13) gene or TSC2 (16p13) gene. Eosinophilic tumour cell clusters appear admixed with macroscopic cysts or microscopic tubules and cystic structures coated with singular layer of hobnail cells. Neoplastic cells are permeated with abundant, granular, eosinophilic cytoplasm. Tumour cells appear immune reactive to PAX8, CK20, AMACR, CD10, SDHB, fumarate hydratase, vimentin, cathepsin K and Melan A.

Infecções fúngicas cutâneas secundárias à varíola aviária no Nordeste do Brasil

ABSTRACT: This study describes the epidemiological and clinical-pathological aspects of outbreaks of avian pox diagnosed in poultry associated with fungal co-infections. A retrospective study was carried out, and cases of avian pox with suspected associated fungal infection were selected. The slides were subjected to routine and special histochemical staining in cases of suspected fungal infection, in addition to performing the immunohistochemical technique. Macroscopically, there were nodular lesions with crust formation, and histologically, the lesions were characterized by hyperplasia of the spinous layer associated with multiple eosinophilic intracytoplasmic inclusions. In seven cases, morphologically compatible structures with fungi were verified through black impregnation with GMS and strongly stained in pink with PAS. The morphotintorial aspects were suggestive of fungi belonging to the genus Aspergillus sp. and Candida sp. The diagnosis of fungal co-infections was confirmed through immunohistochemistry, with positive immunostaining for fungi of the genus Aspergillus sp. (five cases) and Candida sp. (two cases). It is concluded that the occurrence of fungal co-infections secondary to the cutaneous lesions of avian pox represents a complicating factor of the disease, favoring the weakness of these animals and death. Therefore, the investigation of associated secondary agents is necessary.

Identification, molecular characterization, and pathological features of orf virus in sheep and goats in Punjab province, Pakistan

Orf virus (ORFV) causes an acute, contagious, skin disease of sheep and goats which is economically important. The objectives of this study were to identify ORFV and to explore its pathological and phylogenetic profiles in 350 goats and 91 sheep of 14 districts of Punjab, Pakistan, from July 2020 to July 2021. Skin scrapings (total no. of samples = 441) of suspected animals were subjected to polymerase chain reactions, phylogenetic analysis, and pathological observations. The partial length of GIF/IL-2 gene (408bp) was successfully amplified in 58/441 samples. Phylogenetic analysis of GIF/IL2 gene showed that the study isolates belonged to ORFV-cluster I, together with the viruses reported in India and China. Pakistan ORFV isolates were shared 97.6-98.7% nucleotide and 97.6-100% amino acid identities with the reference strain (NC_005336). Moreover, Chinese ORFV-isolates were detected unique multiple amino acid substitutions (F11L, Q21H, D27N, I46V, N49S, N82D, D103N, S129G) with study isolates. Naturally infected animals were anorexic, emaciated, dull, and depressed. The macroscopic lesions included multifocal to coalescing, ulceration followed by proliferative papules, pustules, and crust formation on the epidermis of gums, lips, mouth commissure, muzzles, nose, and udder. Histopathological examination revealed hyperplasia, anastomosing rete ridges formation and degenerative changes, including spongiosis and vacuolation of epidermal cells. Keratinocytes exhibited eosinophilic intracytoplasmic inclusion bodies with pyknotic and karyorrhexis nuclei. This is the first report on molecular characterization of ORFV from Pakistan, with insight into its pathogenesis and comparative analysis of pathological alterations and genetic diversity between ORFV strains reported in different geographical areas.

Eosinophilic homogeneous intracytoplasmic inclusion bodies in acquired epidermodysplasia verruciformis-like lesions.

Eosinophilic homogeneous intracytoplasmic inclusion bodies in acquired epidermodysplasia verruciformis-like lesions.

Clinicopathological and molecular studies on cattle naturally infected with lumpy skin diseases in selected districts of Wolaita Zone, Southern Ethiopia

BackgroundLumpy skin disease is a contagious viral disease of cattle caused by LSDV that results in huge economic losses in the cattle industry. This study characterizes LSDV in cattle through clinicopathological and molecular techniques in selected districts of Wolaita Zone, Southern Ethiopia.MethodsA crossectional study was conducted from November 2020 to June 2021 using Real-time polymerase chain reaction and Histopathological techniques to confirm LSDV.ResultThis study revealed that the percentage of positivity of cattle for LSDV was 36.2%. Clinically, cattle infected with LSDV revealed fever (39–41 °C), nodular lesions on the skin and mucous membranes, and lymphadenopathy. Histopathologically, affected tissue revealed ballooning degenerations of the epidermis, infiltration of mononuclear inflammatory cells, vasculitis, and intracytoplasmic eosinophilic inclusion bodies. RT-PCR confirmed that DNA extracts from skin biopsies of virus isolates were positive for LSDV.ConclusionThe present study confirms that LSDV is widely circulating in cattle of selected districts of the Wolaita zone. Thus, effective control measures through regular vaccination and further confirmation of circulating strains of LSDV through detailed molecular analysis should be recommended.

Molluscum Contagiosum Masquerading as Ocular Surface Squamous Neoplasia

A 72-year-old woman was referred with suspicion of ocular surface squamous neoplasia because 2 leukoplakic nodules were noted on the bulbar conjunctiva (Fig A). Medical history was significant for atopic keratoconjunctivitis and atopic dermatitis. The nodules were excised en bloc followed by alcohol epitheliectomy. Unexpectedly, histopathology of the conjunctival tissue (hematoxylin & eosin stain) showed a cup-shaped lesion with hyperplastic epithelium surrounding intracytoplasmic eosinophilic inclusion bodies characteristic of molluscum contagiosum (Fig B).

A natural outbreak of infectious spleen and kidney necrosis virus threatens wild pearlspot, Etroplus suratensis in Peechi Dam in the Western Ghats biodiversity hotspot, India.

A large-scale mortality of pearlspot, Etroplus suratensis was reported from Peechi Dam, an artificial tropical lake made for irrigation and drinking water supply in Kerala, India during 2018. This dam is located in the premises of Western Ghats, recognized as one of the biodiversity hotspots of the world. The objective of this study was to identify the aetiological agent of this large-scale mortality of E. suratensis by systematic diagnostic investigation and identification of the pathogen. Virus isolation was carried out on a species-specific pearlspot fin (PSF) cell line. Infected PSF cells showed cytopathic effects (CPEs) like cell shrinkage, rounding, enlargement, clustering, and subsequent detachment of cells with a high viral titre of 106⋅95 TCID50 ml-1 at 8 days post-inoculation (dpi). Histopathological examination of the fish showed the presence of numerous abnormal enlarged basophilic cells and intracytoplasmic eosinophilic inclusions in the liver. Moreover, transmission electron microscopy (TEM) analysis revealed the presence of large numbers of 125-132nm viral particles in the spleen tissues. PCR amplification and phylogenetic analysis of the major capsid protein (MCP) gene sequence confirmed that the causative agent was infectious spleen and kidney necrosis virus (ISKNV) of the genus Megalocytivirus. The experimental infection recorded 86.7 ± 2.7% mortality in the E. suratensis (body weight 11.01 ± 2.7g; body length 8.01 ± 2.23cm) injected with 1×104⋅25 TCID50 ml-1 ISKNV per fish. Our detailed investigation provided definitive diagnosis of ISKNV in the severe mass mortality event in wild E. suratensis in Peechi Dam, India, adding one more species to expanding host range of ISKNV infection. The high mortality rate of ISKNV infection in pearlspot suggests the perilous nature of this disease, particularly among the wild fish population.

Molecular and pathological screening of canine distemper virus in Asiatic lions, tigers, leopards, snow leopards, clouded leopards, leopard cats, jungle cats, civet cats, fishing cat, and jaguar of different states, India.

The present investigation was conducted to rule out canine distemper (CD) diseases in Indian wild felids (Asiatic lions, tigers, leopards, snow leopards, clouded leopards, leopard cats, jungle cats, civet cats, fishing cat, and jaguar). The collected samples were screened for CD virus (CDV) by histopathology (HP), immunohistochemistry (IHC) and reverse transcriptase-polymerase chain reaction (RT-PCR) targeting H gene and N gene. The HP and IHC of suspected samples portrayed that 22 [11 leopards, 6 lions, 3 tigers, 1 snow leopard and 1 civet cat] out of 129 (17.05%) wild felids were positive for CD. The major pathological consequences were observed in spleen, lung, kidney and brain. The syncytia and intranuclear as well as intracytoplasmic eosinophilic inclusion bodies were seen in CDV infected cells. Although the histopathological lesions in spleen were more specific and consistent, however, the severe demyelinated leukoencephalitis (usually expected in CD infected dog) was not observed in the brain of any Indian wild felids. Conversely, the CDV antigen has been portrayed via IHC in pancreatic islets of Langerhans of tiger species for the first time in this study. Moreover, the concurrent CD and babesiosis has also been observed in a lioness without a usual coffee-coloured urine. The N gene and H gene of CDV isolates were amplified, sequenced and subsequently constructed the phylogenetic tree. The phylogenetic analysis of H gene revealed that the CDV isolates from Indian lion formed separate clade with CDV isolates from Indian dog and Indian palm civet cat. Furthermore, two CDV isolates from Indian tigers formed clade with Onderstepoort vaccine strain and CDV isolates from dogs of Uttar Pradesh, USA and UK. Evidently, CDV is circulating in Indian wild felids and causing diseases in them.

Pathology and Molecular Characterization of Diphtheritic form of Fowl Pox in Chicken: A Case Report

Fowl pox is viral disease affecting the poultry species mainly chicken, turkey and pigeons worldwide. It exists in two forms viz. cutaneous and diphtheritic form. Grossly, laryngeal opening was completely occluded by caseous material and trachea was severely congested. Microscopically, trachea revealed mucosal thickening due to the proliferation of epithelial cells with characteristic intracytoplasmic eosinophilic inclusion bodies. The collected samples were screened for the P4b gene of fowl pox through PCR. Phylogenetic analysis revealed that the current FPV is closely related to the strains isolated from India and Brazil.

Pathology and Occurrence of Swine Pox in an Organized Swine Farm-Bareilly, Uttar Pradesh

A swine pox outbreak occurred in April 2017 at Bareilly, Uttar Pradesh based organized swine production farm with roughly 200 animals. Twelve growing crossbred (Large White Yorkshire and local) animals were found to have skin lesions, which varied in size and were found on the face, ears, periorbital region, legs, ventral abdomen, inguinal region, dorsal and lateral sides, etc. The lesions ranged from papules and pustules that turned into crusty lesions that looked like brownish scabs. Microscopically, infected cells showed ballooning degeneration of epidermal and follicular epithelial cells, mild epidermal hyperplasia, nuclear vacuolation, and margination of chromatin. Multiple typical eosinophilic intracytoplasmic inclusion bodies were also seen in the epidermal cells, and inflammatory cells were infiltrating the dermis.

Epidemiology, Haemato-biochemical and Pathological Changes Related to Field Outbreaks of PPR in Small Ruminants in Odisha

Background: Odisha experiencing sporadic outbreaks of Peste des petits ruminants (PPR) throughout the year. There is a scarcity of available literature on PPR in Odisha till today. This is the first ever detail investigative approach in the state undertaken with an objective to corelate the epidemiological risk factors, haemato-biochemical and pathological changes in natural field outbreaks occurring in eight different districts. Methods: Fourteen field outbreaks of PPR were evaluated clinically as well as epidemiologically and confirmed through polymerase chain reaction (PCR). Blood, serum, faecal and tissue samples were collected to observe haemato-biochemical and pathomorphological changes to asses disease severity. Result: Present study concluded an overall mortality rate of 46.81%. Chi-square analysis revealed significant highest prevalence among 7-12 months (46.13%) age, Ganjam breed (45.51%) and females (80.49%). Frequent migration among the border areas along with poor management and helminthic infection was major precipitating factor. There was polycythemia along with neutrophilia and lymphopenia. Significant increase in alanine transaminase (ALT), aspartate aminotransferase (AST), K+ and Ca+2 along with creatinine, urea and blood urea nitrogen (BUN) BUN was observed in affected flocks. Antero-ventral consolidation of lungs, syncytia and presence of both eosinophilic intranuclear and intracytoplasmic inclusion bodies were major pathological changes.

Isolation of Lumpy Skin Disease Virus Isolated from SPPV Vaccinated Cattle

Massive outbreaks of suspected LSDV in cattle population were observed during 2017 and 2018 at Beni-Suef governorate. Samples from diseased cattle, previously SPPV vaccinated with SPPV, were inoculated in embryonated chicken eggs (ECE) using chorio-allantoic membrane (CAM) route. Histopathological examination of the inoculated CAMs showed large eosinophilic intracytoplasmic inclusion bodies characteristic for LSDV. The inoculated CAMs were hemorrhagic with congestion blood vessels appeared by the 1st passage then become more pronounced after the second - fourth passages. Characteristic pock lesions were observed after the 1st passage and become clear after the third passage and become clearly observed 4 days post inoculation. For molecular identification, DNA was extracted from a pool of the infected CAMs. Two pairs of primers specific for LSDV including one flanking a 554 bp product of the G gene and second flanking 172 of the RP030 gene were used. In conclusion, LSDV infections have been detected and the virus has been isolated and identified by PCR from cattle previously vaccinated with SPPV vaccine during the summer of 2018.

Histologic lesions of experimental infection with Lymantria dispar multicapsid nucleopolyhedrovirus and Lymantria dispar cytoplasmic polyhedrosis virus in European gypsy moth caterpillars (Lymantria dispar dispar).

European gypsy moths (Lymantria dispar dispar) are an invasive species in North America, and are listed by the International Union for the Conservation of Nature as one of the 100 most destructive invasive species worldwide. They have several known viruses, some of which are used as biological control agents. However, there are no detailed descriptions of many entomopathogenic viral infections, including in European gypsy moths, using bright-field microscopy. In this study, 11 European gypsy moth caterpillars were evaluated histologically: 4 were experimentally infected with Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV; Baculoviridae); 4 were experimentally infected with Lymantria dispar cytoplasmic polyhedrosis virus (LdCPV; Reoviridae); 3 control animals were uninfected. A complete tissue set was evaluated in all animals from all groups using bright-field microscopy, including epidermis, cuticle, striated muscle, tracheae, foregut, midgut, hindgut, Malpighian tubules, hemocytes, fat body, and nervous system. LdMNPV-infected caterpillars had marked karyomegaly and intranuclear viral inclusions in cells of the epidermis, tracheae, fat body, and hemocytes. LdMNPV-infected caterpillars also had hyperplasia and hypertrophy of epidermal and tracheal epithelial cells. LdCPV-infected caterpillars had numerous granular eosinophilic intracytoplasmic viral inclusions in midgut epithelial cells. Both LdMNPV-infected and LdCPV-infected caterpillars had atrophy of fat body adipocytes; this change was more pronounced in LdCPV-infected caterpillars. This work provides the first detailed descriptions of these viral infections in European gypsy moth caterpillars using bright-field light microscopy and provides images of normal histology from control caterpillars.