Herpetic Pneumonia in Indian Ringneck Parrots (Psittacula krameri): First Report of Novel Psittacid Alphaherpesvirus-5 Infection in Europe

Simple Summary Herpesviridae is a large family of double-stranded DNA viruses that infect many different animal species. Herpesvirus infections are common in avian species and, to date, three different alphaherpesvirus species have been recognized as causative agents of disease in Psittaciformes. However, there are reports of respiratory disease in parrots characterized by the presence of distinctive herpes-related histologic lesions, albeit with no identified etiology. Our study acknowledges the unprecedented presence of the novel Psittacid alphaherpesvirus-5 in Europe. Necropsy was performed on Indian ringneck parrots deceased after severe respiratory distress and diagnosis was achieved through histological examination, visualization of the virions by electron microscopy and genome sequencing. The pathogen has been reported only once in Australia and the present report raises the probability that its distribution is wider and it should be included in the list of pathogens threatening parrot populations. The first two European outbreaks of herpetic pneumonia caused by Psittacid alphaherpesvirus-5 were diagnosed based on gross pathology findings, histological examination, transmission electron microscopy visualization and genome sequencing. The outbreaks, characterized by high morbidity and high mortality rates, involved two parrot species, namely the Indian ringneck parrot (Psittacula krameri) and the Alexandrine parakeet (Psittacula eupatria). Clinical signs observed were ruffled feathers, dyspnea, tail bobbing, open wings while breathing, depression and anorexia. Necropsy was performed on Indian ringneck parrots only, and the most evident and serious gross lesion found in all the birds was a diffuse marked consolidation of the lungs associated with parenchyma congestion and oedema. Histological examination confirmed the existence of bronchopneumonia characterized by the presence of syncytial cells with intranuclear inclusion bodies. In one bird, fibrinous airsacculitis was observed as well. Lung tissue inspection through electron microscopy revealed the presence of virus particles resembling herpesviruses. Viral DNA was extracted, amplified using primers for Alloherpesviridae DNA polymerase gene detection, and then sequenced. BLAST analysis showed a 100% identity with the only previously reported sequence of PsHV-5 (MK955929.1)

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Phosphor-converted white light-emitting diodes for indoor illumination need to be warm-white (i.e., correlated color temperature \u3c4000 \u3eK) with good color rendition (i.e., color rendering index \u3e80). However, no single-phosphor, single-emitting-center-converted white light-emitting diodes can simultaneously satisfy the color temperature and rendition requirements due to the lack of sufficient red spectral component in the phosphors’ emission spectrum. Here, we report a new yellow Ba0.93Eu0.07Al2O4phosphor that has a new orthorhombic lattice structure and exhibits a broad yellow photoluminescence band with sufficient red spectral component. Warm-white emissions with correlated color temperature 80 were readily achieved when combining the Ba0.93Eu0.07Al2O4 phosphor with a blue light-emitting diode (440–470 nm). This study demonstrates that warm-white light-emitting diodes with high color rendition (i.e., color rendering index \u3e80) can be achieved based on single-phosphor, single-emitting-center conversion

Cutaneous angiomatosis-like presentation in koi carp (Cyprinus carpio koi): Clinical–pathological investigations

The skin represents an indicator of an animal's health status. Causes of cutaneous diseases in fish most often trace back to biological agents. However, fish skin diseases can also arise from a complex interaction of infectious and non-infectious causes, making it more difficult to identify a specific aetiology. In the period between April and September of the years 2019–2022, four koi carp (Cyprinus carpio koi) from two European countries presented with multifocal, irregularly round, few mm to 1 cm, variably raised cutaneous reddened areas. The fish displayed good general condition. Cutaneous samples, investigated by microbiological and molecular methods and microscopy, did not indicate a primary pathogenic agent. Gross and histological findings of the cutaneous biopsies were consistent with a multifocal/reactive process centred on dermal vessels. The histological features were reminiscent of angiomatosis, a benign proliferative condition affecting the dermal vessels of mammals, including human patients. The clinical–pathological presentation and the dermatologic condition that affected the koi carp are discussed and compared with the veterinary and human literature

Protective Efficacy of H9N2 Avian Influenza Vaccines Inactivated by Ionizing Radiation Methods Administered by the Parenteral or Mucosal Routes

H9N2 viruses have become, over the last 20 years, one of the most diffused poultry pathogens and have reached a level of endemicity in several countries. Attempts to control the spread and reduce the circulation of H9N2 have relied mainly on vaccination in endemic countries. However, the high level of adaptation to poultry, testified by low minimum infectious doses, replication to high titers, and high transmissibility, has severely hampered the results of vaccination campaigns. Commercially available vaccines have demonstrated high efficacy in protecting against clinical disease, but variable results have also been observed in reducing the level of replication and viral shedding in domestic poultry species. Antigenic drift and increased chances of zoonotic infections are the results of incomplete protection offered by the currently available vaccines, of which the vast majority are based on formalin-inactivated whole virus antigens. In our work, we evaluated experimental vaccines based on an H9N2 virus, inactivated by irradiation treatment, in reducing viral shedding upon different challenge doses and compared their efficacy with formalin-inactivated vaccines. Moreover, we evaluated mucosal delivery of inactivated antigens as an alternative route to subcutaneous and intramuscular vaccination. The results showed complete protection and prevention of replication in subcutaneously vaccinated Specific Pathogen Free White Leghorn chickens at low-to-intermediate challenge doses but a limited reduction of shedding at a high challenge dose. Mucosally vaccinated chickens showed a more variable response to experimental infection at all tested challenge doses and the main effect of vaccination attained the reduction of infected birds in the early phase of infection. Concerning mucosal vaccination, the irradiated vaccine was the only one affording complete protection from infection at the lowest challenge dose. Vaccine formulations based on H9N2 inactivated by irradiation demonstrated a potential for better performances than vaccines based on the formalin-inactivated antigen in terms of reduction of shedding and prevention of infection

Ovarian mixed germ cell\u2013sex cord\u2013stromal tumour in a European seabass, Dicentrarchus labrax (Linnaeus, 1758)

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Silent Infection of Highly Pathogenic Avian Influenza Virus (H5N1) Clade 2.3.4.4b in a Commercial Chicken Broiler Flock in Italy

From October 2021 to January 2022, different incursions of clade 2.3.4.4b H5N1 HPAIV (Highly Pathogenic Avian Influenza Virus) occurred in several Italian regions with its main diffusion in Densely Poultry Populated Areas (DPPAs) of north-eastern Italy. Monitoring and control activities applied in the affected area clearly evidenced that turkeys and broilers were the most affected species, although several flocks of broilers at times resulted HPAIV H5N1 infected in absence of increased mortality and/or clinical signs. Thus, an approach based on sampling dead birds was adopted in the broiler sector to improve the early detection of infection; this protocol allowed us to confirm that 15 farms were HPAIV-infected with birds ready to be delivered to the slaughterhouse. The aim of this report is to describe the results of the diagnostic activities carried out in one HPAIV H5N1-infected broiler farm, three days after laboratory confirmation during the pre-movement testing without showing increased mortality or clinical signs. Thus, clinical signs, daily cumulative mortality rate (CMR), virus shedding, seroconversion, pathobiology of clade 2.3.4.4b H5N1 HPAIV as well as Avian Influenza Viruses (AIVs) environmental contamination were thoroughly examined in the infected holding. Such in-depth investigation demonstrated low infection prevalence in live birds, low environmental contamination, no seroconversion for AIVs, gross and microscopic findings compatible with systemic infection with peracute death in H5N1 HPAIV-infected birds

Histological and immunohistochemical characterization of 17 gonadal tumours in koi carp (Cyprinus carpio koi)

Reports on abdominal tumours in koi carp are scarce and most are from the gonads. Their histological diagnosis is challenging due to the occurrence of mixed populations of neoplastic cells and the few availability of cross-reactive antibodies in fish tissues. The present study aims to provide a histopathological characterization of seventeen gonadal tumours, enriched by a wide antibody panel (vimentin, CD117, placental alkaline phosphatase\u2014PLAP, AE1/AE3 cytokeratin, E-cadherin, proliferating cell nuclear antigen\u2014PCNA, m\ufcllerian-inhibiting substance\u2014MIS, GATA4 and Inhibin-\u3b1) applied on whole and tissue microarray (TMA) sections. Abdominal enlargement was associated with tumours filling 30%\u201380% of the abdominal cavity; frequently, the gonads had been completely replaced by neoplastic tissue. Twelve cases were characterized as sex cord-stromal tumours (SCSTs), three as germ cell tumours (GCTs), one as mixed germ cell sex cord-stromal tumour (MGCSCST) and one as carcinoma. By immunohistochemistry, PLAP enabled confirmation of GCTs, ovarian carcinoma and the objective identification of a further cell component in 8 out of the 12 SCSTs that were reclassified as mixed tumours. The use of an immunohistochemical panel can help in refining the histological diagnosis, but the morphological diagnosis still represents the main tool for the characterization of these tumours in koi carp

Cutaneous angiomatosis in koi carp: clinico-pathological investigations and ongoing insights

In the period April-September of 2019 and 2020, five koi carp from different places (Belgium, The Netherlands and Italy) presented with multifocal, irregularly round, few mm to 1 cm, variably raised cutaneous reddened areas, located on the dorsal and ventral body aspects, without any apparent association with coloured areas of the skin. The fish displayed good general conditions, normal behaviour and appetite. After 7-12 months, the skin lesions underwent remission in three out of five carp regardless the treatments. Carp were reared in ponds supplied with water at temperature not controlled, ranging between 18 °C and 25 °C; carp of two ponds were treated with drugs against gyrodactylosis. A clinical-pathological investigation was performed to study the skin lesions. Cutaneous swabs were cultured at 28 °C for 36 hours on blood agar. Skin biopsies were obtained and preserved in buffered formalin and/or alcohol ≥96%. In addition to routine Hematoxylin & Eosin stain to study the morphology of the skin lesions, special stainings (Gram, acid-fast, Warthin-Starry) to investigate potential bacterial agents were performed. Molecular analyses for koi herpesvirus, carp edema virus, Cyprinid herpesvirus 1, Rickettsia-like organism were performed; the samples were also tested with 16S universal bacterial primers. Based on the molecular analyses, viral and bacterial fish pathogens were not found. Bacterial cultures from swabs were negative for primary pathogenic bacteria. Skin parasites or fungi were not detected histologically. The dermis presented a multifocal inflammatory process focused in the dermis characterised by well demarcated round to oval clusters measuring 30-400 μm in diameter, composed of densely packed vascular structures, occasionally forming whorls and inconstantly surrounded by fibrosis. The vascular spaces were occupied by a florid reduplication of small capillaries with aggregates of plump to elongate cells resembling endothelium or pericytes without atypia. In hypocellular areas, a continuum of the lesions could be appreciated, ranging from irregular clusters of plump cells to well-developed micro-vascular structures. Gross and histological findings, common to all cases, were consistent with a process centred on dermal vessels and morphologically consistent with angiomatosis reported in other species. Further morphological investigations are tentatively ongoing in order to elucidate the origin of proliferating vascular/perivascular structures