RESUMO
BACKGROUND: Few cases of scedosporiosis have been reported in animals, but the true prevalence is probably underestimated due to a lack of awareness. Scedosporiosis in dogs has often been associated with localized infection (i.e., nasal infection, eumycetoma, or keratomycosis) or, in rare cases, disseminated infections. CASE PRESENTATION: This case report describes the clinical and pathological features and the diagnostic process of a rare systemic and fatal fungal infection in a dog caused by Scedosporium apiospermum. A 10-month-old female Maremmano-Abruzzese sheepdog showing weakness, lethargy, lateral decubitus, miosis and muscular rigidity was presented. Rodenticide poisoning was clinically suspected for the differential diagnosis. However, postmortem examinations revealed the presence of a swollen and soft subcutaneous nodule located near the right inguinal breast, which was associated with massive enlargement of the inguinal lymph nodes and small disseminated, cream-colored nodules in the kidneys and mesentery. Multiple fungal pyogranulomas were observed upon histological examination. Fungal isolation from the kidneys, breast and inguinal lymph nodes was performed. The internal transcribed spacer (ITS) sequences from the fungal colony DNA were searched in BLAST in the NCBI GenBank for species identification. The sequences of the fungi isolated from the kidney and breast cultures showed 100% sequence identity with sequences from Scedosporium apiospermum. CONCLUSIONS: This report shows that Scedosporium apiospermum may act as a primary pathogen in young and apparently healthy dogs and represents an important pathogen that should be considered during the diagnostic process, particularly when a fungal infection is suspected.
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Doenças do Cão/microbiologia , Infecções Fúngicas Invasivas/veterinária , Scedosporium/isolamento & purificação , Animais , DNA Fúngico , Cães , Feminino , Granuloma Piogênico/microbiologia , Linfonodos/microbiologia , Micoses/veterinária , Scedosporium/genéticaRESUMO
The identification of avian poxvirus and avian papillomavirus associated with cutaneous lesions in griffon vultures (Gyps fulvus) by histopathology, electron microscopy and PCR analysis is reported. Sequence analysis of the fpv140 gene revealed 99% identity to two poxviruses obtained from a white-tailed sea eagle (Haliaeetus albicilla) and a common buzzard (Buteo buteo). Partial sequence of the papillomavirus L1 gene showed sequence similarity to papillomavirus LI genes from African grey parrot (Psittacus erithacus) (69% identity), duck (Anas platyrhynchos) (68% identity), and yellow-necked francolin (Francolinus leucoscepus) (66% identity). To date, this is the first identification of avian poxvirus and papillomavirus in griffon vultures and the first evidence of infection of both viruses in live wild birds.
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Avipoxvirus/isolamento & purificação , Doenças das Aves/virologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/veterinária , Infecções por Poxviridae/veterinária , Animais , Avipoxvirus/genética , Doenças das Aves/epidemiologia , Falconiformes , Variação Genética , Itália/epidemiologia , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Filogenia , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/virologiaRESUMO
Canine prostatic carcinoma is a relevant model for human prostatic carcinoma. Survivin is proposed as a biomarker of malignancy in human prostatic cancer. Sox9 is a stem cell marker required for prostate development and expressed in several adult tissues. The aims of the present study were to evaluate the patterns and expression levels of 2 putative stem cell markers, survivin and Sox9, in canine benign prostatic hyperplasia (BPH) and prostatic carcinoma to investigate their potential as stem cell markers. Immunohistochemistry with specific antibodies was performed on 3 samples of normal prostate gland, 18 samples of canine BPH, and 16 samples of prostatic carcinoma. The basal cell layer of normal and hyperplastic prostatic lobules had nuclear Sox9 immunolabeling and nuclear and rarely cytoplasmic survivin immunostaining, identifying them as potential stem cell markers. Significantly more frequent survivin and Sox9 expression (≥10% of nuclei) was observed in prostatic carcinoma as compared with BPH. The potential coexpression of survivin with Sox9, androgen receptor, and p63 was also investigated in selected BPH and prostatic carcinoma cases with immunofluorescence, and a partial colocalization was observed. Results indicate that Sox9 and survivin could be considered markers of stemness in canine prostate cells. Given its role in proliferation, cells in the basal cell layer with nuclear survivin expression are likely to be transit-amplifying cells that maintain some stem cell proprieties.
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Doenças do Cão/metabolismo , Próstata/metabolismo , Neoplasias da Próstata/veterinária , Fatores de Transcrição SOX9/metabolismo , Células-Tronco/metabolismo , Survivina/metabolismo , Animais , Biomarcadores Tumorais/metabolismo , Doenças do Cão/diagnóstico , Doenças do Cão/patologia , Cães , Imunofluorescência/veterinária , Masculino , Próstata/patologia , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologiaRESUMO
BACKGROUND: Recent investigations have highlighted the controversial role of Wnt/ß-catenin pathway activation in human cutaneous melanoma. Survivin has been proposed as a valid prognostic marker for invasive and metastatic melanomas and lymph node melanoma metastasis in human cutaneous melanoma and is a promising therapeutic target. HYPOTHESIS/OBJECTIVES: Our aim was to investigate the immunohistochemical expression of survivin and ß-catenin in canine cutaneous melanocytic tumours, in order to understand their prognostic significance. METHODS: Twenty-one melanocytic tumours (10 melanocytomas and 11 melanomas) were investigated by immunohistochemistry using specific anti-survivin and anti-ß-catenin antibodies. A semi-quantitative method was used to analyse the results; ß-catenin immunolabelling in neoplastic cells was evaluated as cytoplasmic, membranous or nuclear. The number of survivin-positive cells was counted within ~1000 neoplastic cells. Results were related to histopathological features, evaluated in haematoxylin- and eosin-stained slides, and to the clinical data obtained through a telephone survey with referring veterinarians. RESULTS: Despite a low level of expression in the majority of cases, ß-catenin was found to be correlated strongly with malignant behaviour (P < 0.01). An overexpression of nuclear survivin was statistically related to histological features of malignancy, presence of metastasis and death related to melanoma spread (P < 0.01). CONCLUSIONS AND CLINICAL IMPORTANCE: The low nuclear ß-catenin expression, mainly found in metastatic cases, would indicate that ß-catenin activation may have only limited importance in the development or progression of canine cutaneous melanoma. The correlation of nuclear survivin expression with malignancy would indicate that survivin is possibly a useful prognostic marker and therapeutic target in canine melanoma patients.
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Doenças do Cão/patologia , Melanoma/veterinária , Proteínas de Neoplasias/metabolismo , Neoplasias Cutâneas/veterinária , beta Catenina/metabolismo , Animais , Cães , Feminino , Masculino , Melanoma/patologia , Metástase Neoplásica , Estudos Retrospectivos , Pele/patologia , Neoplasias Cutâneas/patologiaRESUMO
Here we investigated the virulence properties of a unique cell-adapted SARS-CoV-2 mutant showing a ten-amino acid deletion encompassing the furin cleavage site of the spike protein (Δ680SPRAARSVAS689; Δ680-689-B.1) in comparison to its parental strain (wt-B.1) and two Delta variants (AY.122 and AY.21) of concern. After intranasal inoculation, transgenic K18-hACE2 mice were monitored for 14 days for weight change, lethality, and clinical score; oral swabs were daily collected and tested for the presence of N protein subgenomic RNA. At 3 and 7 dpi mice were also sacrificed and organs collected for molecular, histopathological, and immune response profile investigations. The Δ680-689-B.1-infected mice exhibited reduced shedding, lower virulence at the lung level, and milder pulmonary lesions. In the lung, infection with Δ680-689-B.1 was associated with a significant lower expression of some cytokines at 3 dpi (IL-4, IL-27, and IL-28) and 7 dpi (IL-4, IL-27, IL-28, IFN-γ and IL-1α).
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COVID-19 , Interleucina-27 , Melfalan , gama-Globulinas , Camundongos , Animais , Furina/genética , Interleucina-4 , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/genética , Virulência , Camundongos Transgênicos , Modelos Animais de DoençasRESUMO
Mesenchymal stem cells (MSCs) are an important cell population in the bone marrow microenvironment. MSCs have the capacity to differentiate in vitro into several mesenchymal tissues including bone, cartilage, fat, tendon, muscle, and marrow stroma. This study was designed to isolate, expand, and characterize the differentiation ability of sheep bone marrow-derived MSCs and to demonstrate the possibility to permanently express a reporter gene. Bone marrow was collected from the iliac crest and mononuclear cells were separated by density gradient centrifugation. Sheep MSCs cell lines were stable characterized as CD44+ and CD34- and then transfected with a green fluorescent protein (GFP) reporter gene. The GFP expression was maintained in about half (46.6%) of cloned blastocysts produced by nuclear transfer of GFP+ sheep MSCs, suggesting the possibility to establish multipotent embryonic cells' lines carrying the fluorescent tag for comparative studies on the differentiation capacity of adult stem cells (MSCs) versus embryonic stem cells. We found that sheep MSCs under appropriate culture conditions could be induced to differentiate into adipocytes, chondrocytes, and osteoblast lineages. Our results confirm the plasticity of sheep MSCs and establish the foundation for the development of a pre-clinical sheep model to test the efficiency and safety of cell replacement therapy.
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Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Adipócitos/citologia , Adipócitos/fisiologia , Animais , Antígenos CD34/genética , Blastocisto/citologia , Blastocisto/fisiologia , Células da Medula Óssea/fisiologia , Diferenciação Celular , Linhagem da Célula , Centrifugação com Gradiente de Concentração , Condrócitos/citologia , Condrócitos/fisiologia , Feminino , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Receptores de Hialuronatos/genética , Células-Tronco Mesenquimais/fisiologia , Técnicas de Transferência Nuclear , Oócitos/citologia , Oócitos/fisiologia , Osteoblastos/citologia , Osteoblastos/fisiologia , Ovinos , Carneiro Doméstico , Coloração e Rotulagem/métodosRESUMO
BACKGROUND: Epithelial-to-mesenchymal transition (EMT) is a multistep process, important in tumour invasion and metastasis, characterized by loss of epithelial markers, redistribution of ß-catenin and gain of mesenchymal markers. HYPOSTHESIS/OBJECTIVES: Our aim was to investigate the immunohistochemical aberrant expression of cytokeratin, vimentin, survivin and heat shock protein 72 (Hsp72) in canine cutaneous epithelial tumours, to understand the association of expression of these molecules with features of malignancy and their role in the EMT phenotype. METHODS: Ten canine squamous cell carcinomas (SCCs; one with lymph node metastasis), 30 canine hair follicle tumours (six pilomatricomas, eight infundibular keratinizing acanthomas, six trichoepitheliomas and 10 trichoblastomas) and five normal skin samples were investigated by immunohistochemistry using specific anti-vimentin, -cytokeratin, -survivin and -Hsp72 antibodies. A semi-quantitative method was used to analyse the results, as follows: 0 to <5%; ≥ 5 to <10%; ≥ 10 to <25%; and ≥ 25% of positive cells. Immunofluorescence was performed to investigate survivin-vimentin and survivin-Hsp72 colocalization in selected SCCs. Results - In malignant hair follicle tumours and SCCs, a reduced intensity of cytokeratin and increased survivin and Hsp72 expression were observed. In SCCs, loss of cytokeratin expression and vimentin immunolabelling, suggestive of the EMT phenotype, were evident in <5% of neoplastic cells in the front of tumour invasion. In the same areas, strong nuclear survivin and cytoplasmic Hsp72 staining was evident, often colocalizing. Only a few neoplastic cells in the front of tumour invasion showed vimentin-survivin colocalization. CONCLUSIONS AND CLINICAL IMPORTANCE: A possible simultaneous involvement of survivin and Hsp72 in tumour invasion and the multistep process of EMT of cutaneous epithelial tumours of dogs is suggested.
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Transição Epitelial-Mesenquimal/fisiologia , Neoplasias Cutâneas/veterinária , Animais , Diferenciação Celular , Doenças do Cão , Cães , Feminino , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Masculino , Estudos Retrospectivos , Neoplasias Cutâneas/metabolismoRESUMO
Heat shock proteins 70 kDa (HSP70) and apoptosis were investigated in thymus of sea bass juveniles (Dicentrarchus labrax) subsequently to a vaccination against Listonella (syn. Vibrio) anguillarum. HSP70 expression was measured by immunohistochemistry and immunoenzymatic methods, resulting in increase in HSP70 after bath immunization and persistent in fish exposed to an intraperitoneal (i.p.) booster. The HSP70 increase in thymus was suggested as induction in lymphocytic cells, to be related to immune system stimulation after vaccination. However, a thymic recruitment of lymphocyte subpopulations, characterized by higher expression of HSP70, was also hypothesized after vaccination. No apparent relationships were found between HSP70 and apoptosis. In fact, the vaccination did not modulate the apoptosis response, as measured by TUNEL assay and by immunohistochemistry for active caspase-3 expression. The lack of apoptosis effects could be ascribed to the use of inactivated bacteria that appeared not able to interfere with programmed cell death mechanisms. This manuscript aims to contribute to the knowledge of some biochemical features underlying the immunization, with a particular emphasis on the modulation of HSP70. However, further parameters involved in innate/adaptative immunity and apoptosis pathways have to be taken into account to well establish the functional role of HSP70 in fish vaccination.
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Apoptose/imunologia , Vacinas Bacterianas/farmacologia , Bass/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Listonella/imunologia , Timo/metabolismo , Análise de Variância , Animais , Western Blotting , Caspase 3/metabolismo , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Marcação In Situ das Extremidades CortadasRESUMO
West Nile virus (WNV) and Usutu virus (USUV), two antigenically related flaviviruses co-circulating in Europe, can cause severe neurological disease in animals and humans. The immune response against USUV and WNV and their immunopathogenesis are still poorly investigated. Here we present results upon sequential infections of adult immunocompetent CD-1 and BALB/c mice primed with two different doses (high dose, HD or low dose, LD) of an USUV isolate and challenged with HD or LD of three different WNV isolates. CD-1 and BALB/c LD USUV-primed mice, regardless of the dose, are largely protected from lethal WNV challenges despite showing no detectable neutralizing antibodies. Furthermore, mice immunized with a chimeric virus harboring the E protein of USUV within the WNV backbone (WNVE-USUV) are protected against a lethal challenge with WNV. We believe these findings could contribute to understanding the dynamics of the interaction during sequential infection of these two flaviviruses.
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Infecções por Flavivirus , Flavivirus , Febre do Nilo Ocidental , Vírus do Nilo Ocidental , Humanos , Animais , Camundongos , Febre do Nilo Ocidental/prevenção & controle , Febre do Nilo Ocidental/veterinária , Infecções por Flavivirus/prevenção & controle , Infecções por Flavivirus/veterinária , Imunização/veterinária , Anticorpos AntiviraisRESUMO
BACKGROUND: Osteosarcoma (OSA) represents the most common canine primary bone tumour. Despite several pathways have been investigated so far, few molecules have been identified as prognostic tools or potential therapeutic targets, and there is still the need to find out molecular pathways with specific influence over OSA progression to facilitate earlier prognosis and treatment.Aims of the present study were to evaluate the immunohistochemical pattern and levels of expression of a panel of molecules (survivin, ß-catenin, caspase 3 -inactive and active forms- and p53) involved in cell cycle and apoptosis regulation in canine OSA samples, known to be of interest in the study also of human OSA, and to detect specific relations among them and with histological tumour grade, disease free interval (DFI) and overall survival (OS). RESULTS: Nuclear ß-catenin immunostaining was detected in normal osteoblasts adjacent to the tumour, and in 47% of the cases. Cytoplasmic and/or membranous immunostaining were also observed. Nuclear survivin and p53 positive cells were found in all cases. Moderate/high cytoplasmic ß-catenin expression (≥10% positive cells) was significantly associated with the development of metastasis (P = 0.014); moderate/high nuclear p53 expression (≥10% positive cells) was significantly associated with moderate/high histological grade (P = 0.017) and shorter OS (P = 0.049). Moderate/high nuclear survivin expression (≥15% positive cells) showed a tendency toward a longer OS (P = 0,088). CONCLUSIONS: The present results confirmed p53 as negative prognostic marker, while suggested survivin as a potential positive prognostic indicator, rather than indicative of a poor prognosis. The detection of nuclear ß-catenin immunostaining in normal osteoblasts and the absent/low expression in most of the OSAs, suggested that this pathway could not play a major role in oncogenic transformation of canine osteoblasts. Further studies are needed to confirm these hypotheses.
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Caspase 3/metabolismo , Doenças do Cão/metabolismo , Proteínas Inibidoras de Apoptose/metabolismo , Osteossarcoma/veterinária , Proteína Supressora de Tumor p53/metabolismo , beta Catenina/metabolismo , Animais , Caspase 3/genética , Ciclo Celular , Cães , Extremidades/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/fisiologia , Imuno-Histoquímica , Proteínas Inibidoras de Apoptose/genética , Masculino , Osteossarcoma/metabolismo , Proteína Supressora de Tumor p53/genética , beta Catenina/genéticaRESUMO
BACKGROUND: Sebaceous glands are specialized cutaneous adnexal glands, which work under constant hormonal control to produce sebum. They can give rise to several proliferative lesions, such as hamartoma, hyperplasia and neoplasms (adenoma, epithelioma and carcinoma). Their nomenclature is currently confusing, both in veterinary and in human medicine, owing to the difficulty of differentiating between some of these lesions. METHODS: The present study used immunohistochemistry to determine the expression levels and patterns of survivin and Ki67 in five samples of normal canine skin and 44 cases of canine cutaneous lesions with sebaceous differentiation (10 hamartomas, nine hyperplasia, eight adenomas, eight epitheliomas and nine carcinomas). RESULTS: In normal glands, survivin, as well as Ki67, was expressed in scattered reserve cells. In hamartomas, survivin was more highly expressed than in normal skin, indicating a possible role of this molecule in the pathogenesis of these congenital lesions. In tumours, a moderate or high level of survivin and Ki67 expression (more than two and four and more than two positive cells, respectively) were significantly correlated with a malignant histotype, infiltrative growth and a moderate or high number of mitoses (more than two). CONCLUSIONS AND CLINICAL IMPORTANCE: The level of survivin expression increased with increasing malignancy, designating survivin as a new diagnostic marker in the assessment of malignancy of sebaceous tumours.
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Doenças do Cão/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias das Glândulas Sebáceas/veterinária , Animais , Biomarcadores Tumorais , Cães , Regulação Neoplásica da Expressão Gênica/fisiologia , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Proteínas de Neoplasias/genética , Estudos Retrospectivos , Neoplasias das Glândulas Sebáceas/diagnóstico , Neoplasias das Glândulas Sebáceas/metabolismo , Pele/metabolismoRESUMO
A 15-year-old neutered male mixed breed Domestic Shorthair cat was presented for a rapidly growing, intraoral soft gingival mass on the left mandibular region. The neoplastic tissue consisted histologically of two distinct malignant cell populations: spindle cells arranged in bands and epithelioid cells arranged in cords. A few multinucleated giant cells were scattered among the neoplastic cells. Spindle cells and multinucleated giant cells strongly expressed vimentin while epithelial cells strongly expressed pancytokeratins. On the basis of the histological and immunohistochemical results, a diagnosis of oral carcinosarcoma was made. After 2 months, due to the extent of disease and poor prognosis, the cat was euthanized. Necropsy revealed a markedly enlarged, multilobulated white-pink neoplastic mass that had originated from the left side of the sublingual region and involved the coronoid process of the left mandibular bone. The cut surface of the enlarged left submandibular lymph node was glistening, whitish-tan in colour with a multinodular appearance, suggestive of metastasis and confirmed by histological examination. Oral carcinosarcoma is uncommonly recorded in humans and dogs and, to the best of our knowledge, this is the first reported case in a cat.
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Carcinossarcoma , Doenças do Gato , Doenças do Cão , Humanos , Gatos , Masculino , Cães , Animais , Carcinossarcoma/veterinária , Carcinossarcoma/metabolismo , Doenças do Gato/patologiaRESUMO
Mammary carcinomas are the most common neoplasms observed in women and in female dogs. Canine mammary tumors show epidemiological, clinical, genetic, and prognostic characteristics comparable to human breast cancers. The recent introduction of next generation sequencing (NGS) technologies has greatly improved research and diagnostics for humans, while these new tools still need to be implemented in animal models. In this study we developed and validated an AmpliSeq Panel assay for the identification of BRCA variants in twenty-two different dogs. The amplicon mean coverage was 5499× and uniformity was higher than 98% in all samples. The results of germline single nucleotide variants (SNVs) and insertions/deletions (INDELs) were fully concordant regardless of the types of samples considered (blood, fresh and FFPE tissues). Moreover, despite the high DNA degradation observed in older FFPE blocks (>5 years), the assay allowed full coverage of all amplicons for downstream analyses. We consider the NGS panel developed in this study as a useful tool for expanding information on BRCA genes in the veterinary field and for human health from a comparative oncology perspective.
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Abortion in livestock is a public health burden, and the cause of economic losses for farmers. Abortion can be multifactorial, and a deep diagnostic investigation is important to reduce the spread of zoonotic disease and public health prevention. In our study, a multidisciplinary investigation was conducted to address the cause of increased abortion and lamb mortality on a farm, which detected a co-infection of Listeria monocytogenes and Toxoplasma gondii. Hence, it was possible to conclude that this was the reason for a reduced flock health status and the cause of an increased abortion rate. Furthermore, the investigation work and identification of the L. monocytogenes infection root allowed the reduction of economic loss.
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A case of aural carcinoma with chondroid metaplasia at metastatic foci in an 8-year-old male pug is described. Multiple metastases in both lungs and the right submandibular, parotid, retropharyngeal, cervical and prescapular lymph nodes were detected. Histologically, the skin of the right ear canal appeared to be diffusely infiltrated by cords and nests of neoplastic epithelial cells, showing multifocal contiguity with the overlying hyperplastic squamous epithelium. Most of the carcinomatous cells were arranged in a glandular-like pattern, with formation of lumens containing epithelial cells attached to the peripheral cell layer by elongated intercellular bridges. Scattered foci of keratinization with central accumulations of compact, laminated keratin were also observed, and histochemical stains failed to detect mucinous secretory material. Even though histological and histochemical findings were compatible with a diagnosis of acantholytic squamous cell carcinoma, CAM5.2 immunostaining was detectable in the majority, although not all, neoplastic cells, confirming a diagnosis of poorly differentiated ceruminous gland carcinoma. Pulmonary metastatic nodules revealed multifocal areas of cartilaginous metaplasia with apparent transition of carcinomatous cells to chondroid cells, showing nuclear atypia and focal cytokeratin immunostaining. Carcinomatous cells surrounding chondroid areas also revealed focal vimentin and S100 immunoreactivity. Histological evidence of transition between the two components, as well as the presence of intermediate cells displaying both epithelial and mesenchymal immunohistochemical features, strongly indicated a final diagnosis of carcinosarcoma, in which chondrosarcomatous elements were derived from carcinoma cells.
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Carcinoma/veterinária , Doenças do Cão/patologia , Meato Acústico Externo/patologia , Neoplasias da Orelha/veterinária , Animais , Carcinoma/patologia , Cães , Neoplasias da Orelha/patologia , Neoplasias Pulmonares/secundário , MasculinoRESUMO
Novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are emerging worldwide. Here, we report the complete genome sequences of 13 severe acute SARS-CoV-2 strains belonging to lineage B.1.525 (variant η).
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West Nile virus (WNV) and Usutu virus (USUV) are the two most widespread mosquito-borne flaviviruses in Europe causing severe neuroinvasive disease in humans. Here, following standardization of the murine model with wild type (wt) viruses, we engineered WNV and USUV genome by reverse genetics. A recombinant virus carrying the 5' UTR of WNV within the USUV genome backbone (r-USUV5'-UTR WNV) was rescued; when administered to mice this virus did not cause signs or disease as wt USUV suggesting that 5' UTR of a marked neurotropic parental WNV was not per se a virulence factor. Interestingly, a chimeric virus carrying the envelope (E) protein of USUV in the WNV genome backbone (r-WNVE-USUV) showed an attenuated profile in mice compared to wt WNV but significantly more virulent than wt USUV. Moreover, except when tested against serum samples originating from a live WNV infection, r-WNVE-USUV showed an identical antigenic profile to wt USUV confirming that E is also the major immunodominant protein of USUV.
Assuntos
Flavivirus , Febre do Nilo Ocidental , Vírus do Nilo Ocidental , Regiões 5' não Traduzidas , Animais , Flavivirus/genética , Flavivirus/imunologia , Genoma Viral , Camundongos , Virulência , Febre do Nilo Ocidental/prevenção & controle , Febre do Nilo Ocidental/veterinária , Febre do Nilo Ocidental/virologia , Vírus do Nilo Ocidental/genética , Vírus do Nilo Ocidental/patogenicidadeRESUMO
Feline morbillivirus (FeMV) is a newly discovered paramyxovirus infecting domestic cats and its role in the pathogenesis of feline chronic kidney disease (CKD) has been suggested, however not confirmed. The primary aim of the study was to evaluate the renal damage associated with FeMV infection in cats. In this retrospective study, clinical and clinicopathological data were compared among 14 FeMV naturally infected, 21 CKD and 22 healthy cats. FeMV positive cats had serum chemistry analytes and main urine chemistry results similar to the healthy subjects. FeMV positive cats had significantly decreased urine specific gravity (median 1054, range 1022-1065) and urine creatinine (median 227.23 mg/dL, range 83.02-489.75) when compared with healthy cats (median 1067, range 1040-1080, P < 0.001; median 406.50 mg/dL, range 195.32-575.58; P < 0.001, respectively). Urine protein:creatinine ratio (UPC) results of FeMV and CKD were not different (median 0.20, range 0.08-1.03; median 0.23, range 0.10-0.80, respectively), however UPC results were significantly increased in both groups, if compared with healthy cats (median 0.1, range 0.04-0.250, P < 0.01). Based on clinical data, serum creatinine concentration, urine specific gravity and UPC results, CKD was suspected by clinicians in 3/14 FeMV cats. Urine protein sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in 10/13 (77%) FeMV cats indicated a tubular pattern, with a decrease of uromodulin and an increase in the number and intensity of low molecular weight proteins. FeMV infection can be associated with different grades of renal dysfunction ranging from mild tubular proteinuria with less concentrated urine to azotemia in cats younger than those typically affected by CKD.
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In this study, starting from nucleic acids purified from the brain tissue, Nanopore technology was used to identify the etiological agent of severe neurological signs observed in a cow which was immediately slaughtered. Histological examination revealed acute non-suppurative encephalomyelitis affecting the brainstem, cerebrum, cerebellum, and medulla oblongata, while by using PCR-based assays, the nucleic acids of major agents for neurological signs were not detected. By using Nanopore technology, 151 sequence reads were assigned to Bovine Astrovirus (BoAstV). Real-time RT-PCR and in situ hybridization (ISH) confirmed the presence of viral RNA in the brain. Moreover, using the combination of fluorescent ISH and immunofluorescence (IF) techniques, it was possible to detect BoAstV RNA and antigens in the same cells, suggesting the active replication of the virus in infected neurons. The nearly whole genome of the occurring strain (BoAstV PE3373/2019/Italy), obtained by Illumina NextSeq 500, showed the highest nucleotide sequence identity (94.11%) with BoAstV CH13/NeuroS1 26,730 strain, an encephalitis-associated bovine astrovirus. Here, we provide further evidence of the role of AstV as a neurotropic agent. Considering that in a high proportion of non-suppurative encephalitis cases, which are mostly indicative of a viral infection, the etiologic agent remains unknown, our result underscores the value and versatility of Nanopore technology for a rapid diagnosis when the PCR-based algorithm gives negative results.
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Infecções por Astroviridae/veterinária , Astroviridae/isolamento & purificação , Encéfalo/virologia , Doenças dos Bovinos/virologia , Encefalite Viral/urina , Nanotecnologia/métodos , Animais , Astroviridae/classificação , Astroviridae/genética , Infecções por Astroviridae/diagnóstico , Infecções por Astroviridae/patologia , Infecções por Astroviridae/virologia , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/patologia , Encefalite Viral/diagnóstico , Encefalite Viral/patologia , Encefalite Viral/virologia , Itália , FilogeniaRESUMO
Feline morbillivirus (FeMV) is an emerging morbillivirus first described in cats less than a decade ago. FeMV has been associated with chronic kidney disease of cats characterized by tubulointerstitial nephritis (TIN), although this aspect is still controversial and not demonstrated with certainty. To investigate FeMV prevalence and genomic characteristics, an epidemiological survey was conducted in a total number of 127 household cats originating from two Italian regions, Abruzzi and Emilia-Romagna. A total number of 69 cats originating from three feline colonies were also enrolled for the study. Correlation with TIN was investigated by employing a total number of 35 carcasses. Prevalence of FeMV RNA was higher in urine samples collected from cats of colonies (Pâ¯=â¯31.8%, CI 95% 22.1-43.6) compared to household cats (Pâ¯=â¯8.66%, CI 95% 4.9-14.9) and in young and middle-aged cats while prevalence of FeMV Abs was higher in old cats. Sequences obtained straight from infected biological samples, either partial or complete, cluster into two clades within FeMV genotype 1, distantly related to FeMV genotype 2. Immunohistochemistry analysis of kidney sections of FeMV RNA positive cats revealed immunoreactivity within epithelial cells of renal tubuli and inflammatory cells. However, statistically significant association between FeMV and renal damages, including TIN, was not demonstrated (p= 0.0695, Fisher exact test). By virus histochemistry performed with FeMV-negative feline tissues and a FeMV isolate, tropism for different cellular types such as inflammatory cells residing in blood vessels of kidney and brain, airway epithelial cells, alveolar macrophages and to a lesser extent, the central nervous system, was demonstrated. Additional studies are warranted in order to establish viral tropism and immune response during the early phases of infection and to disentangle the role of FeMV in co-infection processes.