A survey of the canine pantropic coronavirus infection was carried out as part of the EU Canine Pantropic Coronavirus Epidemiological Survey in 2009-2011. The aim was to detect the presence of canine pantropic coronavirus (CPCoV) in Hungarian dog populations. Samples have been taken from a total of 130 dogs, and polymerase chain reaction after reverse transcription (RT-PCR) has been performed. 18 samples from organs and 11 samples from faeces were positive for canine enteric coronavirus (CCoV). Further specific examination of CPCoV with real-time PCR was carried out, 12 from 18 organ samples were positive, while none of the 11 faecal samples. Virus isolation was not successful from the samples. However, from a total of 29 coronavirus positive samples 27 were simultaneously positive for canine parvovirus type 2 (CPV-2), which raises the question if the observed symptoms and lesions were only caused by CPCoV. The data indicate that canine pantropic coronavirus is present in the Hungarian dog populations, and its occurrence is quite high, since 12 of the 130 tested dogs (9.23%) were carrying CPCoV.

Transmissible viral proventriculitis (TVP) is currently a serious and ongoing problem in the poultry industry, causing significant financial losses worldwide. Numerous viral pathogens have been suspected to be the etiological agent of TVP, including birnaviruses, coronaviruses and reoviruses, however until recently no definite causative agent was determined for this disease. In this study chickens from 13 commercial Romanian flocks with increased mortality rate, feathering problems and clinical signs suggestive of TVP were examined by histopathology and methods applying polymerase chain reaction (PCR) for the direct demonstration of viruses suspected to play a role in TVP: avian reovirus, infectious bursal disease virus and chicken proventricular necrosis virus (CPNV). Histopathology revealed lesions suggestive of TVP. All the flocks were found positive by PCR for avian reovirus but none were positive for the newly implicated CPNV.

The authors describe the course of an acute disease in a goose flock, affecting two age groups. The stocks were 2 and 3 weeks old geese, and five days after the appearance of the first clinical symptoms 99% of the animals died. The epidemiological investigation revealed inappropriate vaccination of the parent stock so the young geese were unprotected against exposure to the parvovirus, the causative agent of Derzsy's disease. This case draws attention to the importance of the proper vaccination programs in goose flocks and to the mass economical loss due to the failure of protection against the viral agent, which could have been avoided by following the rules of animal health.

Samples collected in 2008 and 2009, from 49 turkey flocks of 6 to 43 days in age and presenting clinical signs of enteric disease and high mortality, were tested by polymerase chain reaction and reverse transcription-polymerase chain reaction for the presence of viruses currently associated with enteric disease (ED) syndromes: astrovirus, reovirus, rotavirus, coronavirus, adenovirus, and parvovirus. Turkey astroviruses were found in 83.67% of the cases and turkey astrovirus 2 (TAst-2) in 26.53%. The investigations directly demonstrated the high prevalence of turkey parvovirus (TuPV) in 23 flocks (46.9%) experiencing signs of ED, making this pathogen the second most identified after astroviruses. Phylogenetic analysis on a 527 base pair-long region from the NS1 gene revealed two main clusters, a chicken parvovirus (ChPV) and a TuPV group, but also the presence of a divergent branch of tentatively named "TuPV-like ChPV" strains. The 23 Hungarian TuPV strains were separately positioned in two groups from the American origin sequences in the TuPV cluster. An Avail-based restriction fragment length polymorphism assay has also been developed for the quick differentiation of TuPV, ChPV, and divergent TuPV-like ChPV strains. As most detected enteric viruses have been directly demonstrated in healthy turkey flocks as well, the epidemiology of this disease complex remains unclear, suggesting that a certain combination of pathogens, environmental factors, or both are necessary for the development of clinical signs.

The major enteric disease (ED) complex in broiler chickens is runting-stunting syndrome and in turkey broilers is poult enteritis mortality syndrome. Viruses from numerous families have been identified in the intestinal tracts of poultry with ED, such as Astroviridae, Coronaviridae, Reoviridae, Rotaviridae, and Parvoviridae. The objective of the present study was to directly demonstrate the presence of the scarcely known chicken parvovirus (ChPV) and turkey parvovirus (TuPV) in Hungarian flocks experiencing clinical signs of ED. ChPV and TuPV infection were demonstrated in 15 chicken flocks and two turkey flocks, in intestinal samples collected between 2008 and 2010. The histopathological investigation revealed enteritis in the duodenum and jejunum, and atrophy of the lymphoid organs. Indirect immunohistochemistry (IHC) suggested the intestinal epithelium of chickens and turkeys as a potential replication site of the virus, similarly to other parvoviruses, while in case of the turkey samples IHC positivity was also observed in the bursa of Fabricius, liver and pancreas. However, no direct connection could be established between the presence of the pathogen in the above-mentioned tissues and the histopathological changes observed in the investigated flocks. The phylogenetic analysis performed on the partial nucleic acid sequence of the NS1 gene revealed an evident clustering tendency of the ChPV and TuPV strains, but also highlighted the potential reciprocal role of these two species in the epidemiology of these viruses. The role of the ChPV and TuPV in the ED is far from understood, but the results of the present study emphasize the fact that in certain, still not fully elucidated conditions, ChPV and TuPV may participate in the emergence of ED in chicken flocks, as suggested by previous experimental infections.

BACKGROUND:Nanoparticles represent promising drug carrier systems. In the case of cytostatics such as doxorubicin, carrier colloid systems as human serum albumin (HSA) nanoparticles, may increase their therapeutic efficiency and decrease their side-effects (toxicity) and any potential multidrug resistance. In the present study, doxorubicin, as a widely used antineoplastic agent, was incorporated into the matrix of human serum albumin and three different particle-sized doxorubicin-loaded HSA nanoparticles were prepared, using a previously described desolvation method. Our objective was to find out if different particle sizes of colloid carriers can allow regarding the given cytostatic agent.MATERIAL AND METHODS:The three prepared nanoparticles were labelled using technetium (Tc-99m) and were tested for their physicochemical colloidal quality, fluctuations, and radiochemical stability. Biodistribution of different-sized radiolabelled colloids were determined by means of scintigraphic imaging studies in healthy male Wistar rats. Images were taken by gamma camera at several times and organ uptakes were estimated by quantitative ROI analysis.RESULTS:In vitro measurements showed that more than 95% of doxorubicin proportion was permanently adsorbed to human serum albumin. Radiolabelled doxorubicin-loaded particles had high-degree and durable labelling efficiency and particle size stability. Biodistribution results had a close correlation to earlier described results of radiocolloids in similar particle size ranges. In vivo examinations verified that colloid carriers have insignificant size fluctuations after an intravenous application and they show the proper distribution according to their particle size.CONCLUSIONS:Our investigations verified that different and stable particle sizes make drug carrier HSA nanoparticles possible to apply different drug targeting in a potential clinical use.

Here we report a case of canine babesiosis with unusual morphology of the causative agent. A male, seven-week-old Labrador retriever puppy, exhibiting severe anaemia and haemoglobinuria, was presented at the Clinic of Internal Medicine in February 2011. The puppy was euthanised. The most relevant pathological changes were icterus, severe splenomegaly, generalised lymphadenopathy and haemoglobin nephrosis. Samples were collected from various organs for histology within one hour post mortem. Impression smears were also prepared from the spleen after overnight storage at 4 °C. Tissue sections and smears showed the presence of multiple, coccoid intraerythrocytic bodies that measured 1-2 μm and resembled small babesiae. No large piroplasms were seen. DNA was extracted from the spleen, and a conventional PCR was performed for the amplification of a 450-bp region of the 18S rRNA gene of piroplasms. The causative agent was identified as Babesia canis canis, with 99% sequence identity to other European isolates. Sequence identity to B. gibsoni was only 91%. This is the first account to verify that the morphology of the large canine piroplasm, B. canis, can be uniformly small babesia-like post mortem or following the storage of tissue samples.

Transmissible viral proventriculitis (TVP) is a poorly understood disease of broiler chickens. It is characterized by enlargement of the proventriculus, retention of ingesta in the lumen, and weakness of the gastric isthmus. Microscopically, degeneration and necrosis of the proventricular glandular epithelium is accompanied by marked lymphocytic infiltration. Samples from 13 chicken broiler farms from Romania, exhibiting clinical signs of TVP, poor feed conversion efficiency, and increased mortality have been examined by necropsy, histopathology (haematoxylin and eosin and Giemsa staining). The histopathological findings were dominated by the presence of enlarged, deformed glands, containing detritus with an inflammatory cell infiltrate composed of lymphocytes, histiocytes and surprisingly mast cells, as determined by special staining.

Avian encephalomyelitis (AE) is a viral disease infecting mainly the brain and spinal cord of young poultry species, especially chickens, but due to the regular vaccination during the last decades, it is very rarely diagnosed in clinical form. In the present article the authors describe an acute case of infectious encephalitis and myelitis resulting in high mortality in two imported broiler flocks, and emphasize the resulted severe economical loss which could have been avoided by obeying the rules of state veterinary medicine.

Feline parvovirus (FPV) induced disease in cats has been known since the beginning of the 20th century. Despite initial observations, it has been demonstrated that lions (Panthera leo) may also be infected and develop clinical disease. The carcass of a 1.5 years old lioness that prior to death showed various clinical signs, including diarrhoea, walking in circles and in the final stages of the diseases refusal of movement and lack of response to external stimuli, was examined for diagnostic purposes. Other three animals that lived with the examined lioness presented similar clinical signs and died. The macroscopic and histopathologic examinations revealed the likelihood of a parvovirus infection with secondary complications (penetrating ulceration of the duodenum, necrotizing myositis, cerebral oedema etc.). A polymerase chain reaction based diagnostic test confirmed the parvovirus infection, while the nucleic acid sequence analysis of the VP2 gene identified the pathogen as an FPV strain more closely related to a German isolate (U22188) than to Hungarian FPV strains. The present work discusses the unfortunate consequences of the neglecting prophylactic immunization of susceptible exotic animals that could be in contact with other, free-roaming animals.

Feline parvovirus (FPV) induced disease in cats has been known since the beginning of the 20 century. Despite initial observations, it has been demonstrated that lions (Panthera leo) can also get infected and develop clinical disease. The present work describes the results of the macroscopic, and light microscopic changes induced by FPV and the electron microscopic and genetic demonstration of the infection in several species, as well as the phylogenetic analysis of FPV strains currently circulating in Hungary.

Type 2 canine parvovirus (CPV2) infection is one of the most frequent causes of death in the young, susceptible canine populations worldwide. Since its emergence in the 1970s, several variants have been described. In the present study the authors describe the genetic analysis of 24 Hungarian CPV2 strains collected from 2004 to 2008. Surprisingly, the genetic and phylogenetic investigations of all these strains revealed that all of them were type 2a CPVs. On the other hand, the genetic analysis provided substantial evidence to demonstrate that due to a seemingly constant point mutation present in most of the Hungarian CPV2a strains, a previously described MboII-based rapid identification of CPV2c strains unfortunately cannot be reliably used any more.

The authors describe a squamous cell carcinoma arising from the ear canal of a Long-eared Hedgehog (Hemiechinus auritus). No metastasis could be identified elsewhere in the animal. Due to the irritation caused by the tumorous proliferation the animal constantly scratched the affected area, which led to secondary bacterial infection of the middle ear accompanied by the stagnation of an increased volume of local secretions. Using routine haematoxylin and eosin and immunohistochemical staining techniques, the tumour was identified as a squamous cell carcinoma. This work constitutes the first description of such a tumour in a Long-eared Hedgehog.

The molecular characterization of 4 Hungarian chicken anaemia virus (CAV) strains retrieved from naturally infected broiler chickens was studied, after the recent re-emergence of chicken infectious anaemia (CA). Carcasses from 4 Hungarian broiler flocks experiencing increased mortality were examined by necropsy, histopathology and polymerase chain reaction (PCR) based technique. The chickens presented stunted growth and general anaemia expressed by pallor and haemorrhages in the subcutaneous tissue. Blue wing disease was found in case of one flock (997/99), affecting 70% of the diseased chickens. Secondary bacterial and fungal infections were identified in several chickens from all investigated flocks. Histological examination revealed hypoplasia of the bone marrow and lymphocyte depletion in the spleen, bursa of Fabricius and thymus. The alignment from the deduced VP1 amino acid (aa) partial sequences (114 aas) showed high similarity (99%) among the Hungarian strains except for one that proved to have a higher variation, presenting 97% identity with the rest of the Hungarian CAV strains. Several aas substitution were identified in the Hungarian strains. Blocks of sequence data leading 344 bps were used for the phylogenetic analysis, and the Hungarian nucleotide sequences were compared with 22 other CAV sequences deposited in the GenBank to investigate any geographical relationship. The Hungarian sequences were not positioned in a separate group of the phylogenetic tree, nor did they cluster together only with European CAV strains. This study confirms the presence of CAV infection in Hungary, and presents the sequence and phylogenetic analysis of the circulating CAV strains.

Canine distemper (CD) is a highly contagious, often fatal, multisystemic viral disease of receptive carnivores. The presence of a PsiI cleavage site on a specific location of the hemagglutinin (H) gene was found to be a hallmark of vaccine strains, thus, a previously published restriction fragment length polymorphism (RFLP) test using PsiI theoretically allows the distinction between all currently used vaccine strains and virulent field strains. The RFLP test was carried out on all brands of CD vaccines available in Hungary. The present work describes the extensive sequencing and phylogenetic study of the strain present in Vanguard (Pfizer Animal Health) vaccines, which following the PsiI based RFLP test reacted as a wild-type strain. Based on the product description provided by the manufacturer, all batches should have contained a virus strain (Snyder Hill) belonging to the group of vaccine strains (America-1). Extensive genetic analysis involving the full nucleic acid sequence of four other genes (N, M, P and F) of the CDV genome revealed that the incriminated virus strain showed a higher level of genetic identity to wild-type strains from the America-2 group than to any of the strains belonging to America-1 group, therefore the vaccine does not contain the virus strain stated by the manufacturer in its product description and has not been containing it since at least 1992.