PCV2 infection in vaccinated conventional gilts inseminated with PCV2b-spiked semen

The present trial investigated the effect of PCV2 vaccination on viremia, virus shedding and viral load in maternal tissues and foetuses of conventional gilts inseminated with PCV2b-spiked semen. Twelve gilts were randomly divided into two groups of six animals each (vaccinated infected, VI; non-vaccinated infected, NVI). Estrus synchronization was followed by artificial insemination (AI) with a single PCV2 negative semen dose supplemented with 0.2 mL of a PCV2b suspension containing 104.4 TCID50/50 \u3bcL (total viral dose: 105 TCID50). Vaginal, nasal and faecal swabs, and blood samples were collected weekly from two days before artificial insemination till the end of the experimental period (55 days post AI; DPAI) and tested by real-time PCR (qPCR) for PCV2; sera were tested for anti- PCV2 antibodies. During necropsy foetal and maternal tissues were collected for qPCR and histopathology. In each of the VI and NVI groups three out of the six gilts were pregnant at 29 DPAI. The VI group showed a significantly lower proportion of PCR-positive swabs: 24.6% VI vs 71.3% NVI. PCV2 clearance was demonstrated by qPCR in lymphoid tissue during the trial in the VI group. Only one foetus was PCV2-positive (in the NVI group) and three amniotic fluids of the NVI group. PCV2 was found in a significantly lower proportion of the placenta of foetuses in the VI group (39%) than the NVI group (77%). The PCV2 vaccine seems to play an active role in reducing virus shedding, tissue viral load and foetoplacental infection

PCV2-DNA in formalin-fixed and paraffin embedded lymph nodes of wild boar (Sus scrofa ssp. scrofa): one sampling approach for two laboratory techniques

Superficial inguinal lymph nodes from 72 wild boars examined in a previous immunohistochemical (IHC) study on porcine circovirus type 2 (PCV2) were selected for a PCV2 polymerase chain reaction (PCR) analysis. Four of these lymph nodes were PCV2-IHC strongly positive with PMWS histological lesions (outcome 1), 6 weak to mild PCV2-IHC positive without PMWS histological lesions (outcome 2) and 62 PCV2-IHC negative. Considering IHC the gold standard for diagnosis, the aims of the study were to evaluate the suitability of the PCV2-DNA extraction from formalin-fixed and paraffin-embedded (FFPE) tissue and the sensitivity and specificity of PCR under two IHC interpretations criteria: (A) the sample was considered positive if the result was outcome 1; (B) the sample was considered positive if the result was outcome 1 or 2. Under (A) criteria, sensitivity and specificity of PCR were 100% and 89.7%, respectively; the Cohen's Kappa coefficient was 0.49. Under (B) criteria, sensitivity and specificity of PCR were 80.0% and 95.2%, respectively; the Cohen's Kappa coefficient was 0.72. The high Cohen's Kappa coefficient under the (B) interpretative criteria indicates good agreement between the two methods. In conclusion, 1) DNA extracted from FFPE specimens of wild boar is suitable for PCR and further represents a screening test for PCV2/PCVD (PCV2 Diseases) investigations in wild boar as well; 2) routine histological sampling can also be useful for PCV2 virological studies in wild boar

Indagine immunoistochimica per circovirus suino tipo 2 in una popolazione di cinghiali (Sus scrofa ssp. Majori) della provincia di Bologna

It is well known that wild boar (Sus scrofa ssp. majori) is a host of porcine circovirus 2 (PCV-2). In many Countries, studies reported the presence of this virus in wild boar. One hundred and sixty-five lymph nodes were sampled from 86 wild boars shot in the province of Bologna. Histologically 3 cases displayed a PMWS related pattern; 1 of which was stained by immunohistochemistry (IHC) (revealing the viral capsid antigen), that also showed 6 positive subjects (7%), as the whole. In situ hybridisation (ISH) (performed in 1 out of 6 reactive cases) was positive, as well. The statistical parameters were not evaluated because of the small population sampled. However, PCV-2 was detected by histology-IHC and ISH combination in wild boar in Emilia-Romagna

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

open6noTelomere length maintenance is regarded as a fundamental step in tumorigenesis, as most human brain tumors, including meningiomas, stabilize the ends of their chromosomes using telomerase. This investigation represents an introduction to telomerase expression in canine and feline meningiomas. Twenty-five archived cases (14 dogs and 11 cats) were immunohistochemically tested for human-telomerase reverse transcriptase (h-TERT), scored, and quantified; furthermore, mitoses were counted on sections stained with a modified toluidine blue. The h-TERT antibody immunolabelled the nucleus and nucleolus of meningeal neoplastic cells, with an intensity ranging from mild to strong and a speckled distribution; a significantly higher expression in cats was noted, while no significant association between h-TERT immunolabelling and sex or histotype was evident in dogs or cats. The telomerase enzyme represents a fundamental parameter of potential malignant transformation, which may occur independently of the signal to proliferate, thereby supplying the cells with unlimited growth capabilities. Telomerase expression could be a prognostic indicator independent of the kinetic parameters, although this should be evaluated using a larger dataset with available clinical information.openL. Mandrioli; S. Panarese; A. Cesari; M.T. Mandara; P.S. Marcato; G. BettiniL. Mandrioli; S. Panarese; A. Cesari; M.T. Mandara; P.S. Marcato; G. Bettin

PCR per PCV2 in linfonodi di cinghiali (Sus scrofa ssp. scrofa) fissati in formalina ed inclusi in paraffina (FFPE): valutazione comparativa con i risultati immunoistochimici.

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Facial myxomatous chondroma in a neon tetra (Paracheirodon innesi)

Neoplasms of mesenchymal origin are generally rare in fish, with the only exception of fibromas and fibrosarcomas. Spontaneous cartilage tumors have been reported occasionally; examples are a chondroma of the gill arch in a brown trout, an osteochondroma of the operculum in a jewel fish, a cranial chondrosarcoma in a paddlefish and ocular chondrosarcomas in mangrove rivulus. Bone neoplasms are also uncommon, and reports include a branchial osteosarcoma in a barbel, a cutaneous osteosarcoma in a channel catfish, a branchial osteochondroma in a gilthead sea bream and multiple osteomas in thinspine sea catfishes. Furthermore, osteosarcomas, chondromas, rhabdomiosarcomas and leyomiosarcomas have been reported in laboratory fishes exposed to N-methyl-N’-nitro-N-nitrosoguanidine (MNNG). Mouth, lips, and jaws are frequent site of neoplasia in fish, presenting as proliferative, nodular exophytic growths, generally resembling papillomas; they have often been histologically classified as fibromas or fibrosarcomas. Advanced, enlarged lesions may eventually result in death of the fish due to compromised feeding and respiration. A three-cm neon fish (Paracheirodon innesi) presented a single, well-circumscribed, nodular, roughly spherical, perlaceous, firm to hard, 4 mm in diameter mass in the right facial region; due to a difficulty in feeding it was euthanized with lethal dose of anaesthetic. The whole fish was fixed in formalin, embedded in paraffin-, and 4-μm sections were cut. From the right maxillar region a nodular, well circumscribed mass, covered by an epithelial layer and formed by abundant, highly cellular cartilaginous tissue and loose myxomatous matrix, was detected. The cartilaginous tissue contained nests of chondrocytes lying in elliptic lacunae, consistent with multicellular isogenous cell groups of mature hyaline cartilage. Beside these lacunae, large cells (50-80 μm in diameter), with distinct cell borders and single large, clear vacuoles (physalipherous cells) were present; nuclei were small, round to oval shaped, hyperchromatic and often eccentrically located. In both these types of cells, nucleoli were only occasionally discernible and mitotic figures were rare. Poorly cellular myxomatous tissue was also present. A diagnosis of facial myxomatous chondroma was made. Histological sections were stained with Alcian-PAS and Masson’s trichrome, in order to demonstrate the presence of proteoglycans and collagen within the neoplastic tissue. As defects in the pathways of cell cycle control have been reported in fish tumors, particularly in the zebrafish, immunohistochemistry was also performed using antibodies against Bcl-2 protein, telomerase reverse transcriptase catalytic subunit (h-TERT) and p53 protein. Indeed, h-TERT may suppress apoptosis interacting with Bcl-2 and p53 proteins. The antibodies tested did not show the immunohistochemical overexpression of any of these markers, which suggests that the breakdown of other mechanisms could be involved in the oncogenesis of this myxomatous chondroma

Telomerase Reverse Transcriptase (h-TERT) immunohistochemistry in canine and feline meningiomas

Telomerase is a ribonucleoprotein complex that prevents the erosion of chromosomal extremities (telomeres) in eukaryotic cells. The role of telomerase in the process of tumoural transformation is currently undergoing intense scrutiny, as its activation is considered a fundamental step in “immortalization” of neoplastic cells, and has been suggested to play a key role in the progression of several tumours, including human intracranial meningiomas. This study aimed to evaluate in 25 cases of archived meningiomas (14 canine and 11 feline) the presence of telomerase, determined by immunohistochemistry (IHC) with an anti-h-TERT monoclonal antibody (clone 44F12, Novocastra), which detects the enzymatic catalytic subunit. The positive reaction, identified by the evidence of scattered nuclear and/or nucleolar staining, was computed with an automated image analysis system ("Lucia 32G/Mutech", Nikon) and expressed as percentage of positive tumoural cells. According to WHO International Classification of Tumors of the Nervous System of Domestic Animals, meningiomas were grouped into the following histotypes: meningothelial (6 canine), transitional/mixed (1 canine, 6 feline), psammomatous (1 canine, 2 feline), fibrous/fibroblastic (1 canine, 1 feline), anaplastic/malignant (5 canine, 2 feline). In 15 out of 25 meningiomas (6 canine, 9 feline) h-TERT protein was localized in the nuclei, notably nucleoli and occasionally in the cytoplasm (two cases) of the cells, with mild to strong staining intensity. Even the percentage of positive cells was variable, ranging from 5 to 80%, regardless of the histological type. Although the number of telomerase-negative meningiomas may depend upon overfixation, the heterogeneous h-TERT expression suggests the existence of alternative mechanisms to telomerase involved in the tumoural transformation and prompts the necessity to correlate h-TERT expression with proliferative activity and biologic behaviour

Evidence of Pneumopericardium after Elective Ovariectomy in a Peritoneopericardial Diaphragmatic Hernia-Affected Dog: A Case Report

Peritoneopericardial diaphragmatic hernia (PPDH) is an opening between the pericardial sac and the pleuroperitoneal membrane. Pneumopericardium is an infrequent complication of PPDH. This condition is a serious circumstance in which free gas accumulates in the pericardial sac. The present report describes the occurrence of pneumopericardium and pericardial effusion after elective ovariectomy in a dog affected by PPDH. The presence of an umbilical and diaphragmatic hernia was highlighted during ovariectomy, and a pneumopericardium was seen during an X-ray exam. At the time of admission to the hospital, the dog was asymptomatic. The diagnosis was performed by X-ray and ultrasonographic exams. Computed tomography examination confirmed the diagnosis and directed for a surgical approach of the congenital defect. Surgery resulted in resolution of PPDH and of pneumopericardium

Porcine circovirus type 2 detection in in vitro produced porcine blastocysts after virus sperm exposure

This study was aimed at assessing the capability of semen experimentally infected with porcine circovirus type 2 (PCV2) to produce porcine blastocysts PCR positive for PCV2. Embryos were obtained from in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes or by parthenogenesis. Sperm suspension was exposed to PCV2b and utilized for IVF. PCV2 spiked semen did not reveal any reduction in sperm viability or motility but its ability to produce infected blastocysts was irrelevant as only one out of 15 blastocysts obtained by IVF were PCV2b; however two blastocysts were PCV2a positive. Furthermore, the presence of PCV2 was demonstrated also in embryos obtained by parthenogenesis (one out of 17 was PCV2b and one PCV2a positive). Even if PCV2 firmly attaches to the surface of spermatozoa, experimentally spiked sperm were not effective in infecting oocytes during IVF and in producing PCR positive embryos. The infected blastocysts we obtained derived most probably from infected oocytes recovered at the abattoir