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The aim of this study was to describe long-term follow-up and difference in immune reactions in the tear film following penetrating keratoplasty (PK) in horses when differently preserved corneas were utilised. This report describes for the first time the use of corneal grafts preserved in tissue culture media in equine PK. Eight experimental horses with normal eyes were included and freshly harvested, frozen or preserved corneal grafts were used for the PK. The graft-taking technique and storage, PK surgery, postoperative treatments and complications are described. The mean postoperative follow-up time was 286 days. Tear film samples taken before and periodically after surgery were measured for IgM, IgG and IgA contents by direct ELISA. All grafts were incorporated into the donor horse but were rejected to some degree. The differently harvested corneal grafts healed in the same manner and looked similar. Preoperatively, the clear corneas meant low risk for graft failure, and the fresh or stored tissues provided intact endothelium, although there were no clear graft sites postoperatively. The presence of IgA, IgG and IgM was demonstrated in the tear film from the early postoperative period. IgG levels were lower than IgA or IgM and had a constant baseline in every case, as IgA and IgM had great variability with time and an individual pattern in each eye.

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Little information is known about infection, replication and transmission of porcine circovirus type 2 (PCV2) in species other than swine. Two sets of animal experiments were carried out to investigate the susceptibility of mice to PCV2 and to study their possible role in maintaining and transmitting the virus. In the first experiment 14 mice were inoculated with PCV2 by the intraperitoneal route with 5 × 10 2 TCID 50 of the PCV2-ROM strain (Cadar et al., 2007). In a second experiment 24 mice were divided into two groups (A and B); mice in Group A (n = 18) were inoculated orally with 1 × 10 5 TCID 50 PCV2-ROM and mice in Group B (n = 6) were left uninoculated until day 12 post inoculation (p.i.), when they were mixed with Group A. The animals were sacrificed at intervals for postmortem investigation and virus genome detection by polymerase chain reaction (PCR). The PCR results indicated that PCV2 could replicate in mice infected intraperitoneally or by the oral route, and that the virus can be transmitted directly from mouse to mouse.

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Porcine circovirus type 1 (PCV1) is considered to be a non-pathogenic virus detected in cell cultures, vaccines or products used for cell culture preparations, all of them of porcine origin. Serological evidence and genetic studies suggested that PCV1 was widespread in domestic pigs. The presence of PCV1 in wild boars in Germany was also described using serological methods. This paper reports the first detection of PCV1 in Hungarian wild boars. Samples were collected at slaughterhouses and processed for polymerase chain reactions. The complete genome of PCV1 detected in the samples was determined and compared with the available PCV1 sequences of the GenBank database. The genomes formed two distinct clusters with minimum differences, where the Hungarian wild boar PCV1 (WB-H8) grouped together with genomes originating from domestic swine from China and Australia and with a genome detected in a porcine pepsin product.

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A recombinant cucumber mosaic virus based expression system has been developed for the production of an immunogenic porcine circovirus epitope. The resulting nanoparticle was shown to elicit specific immune response in mice and pigs, when administered parenterally. To evaluate the oral applicability of this vaccine candidate, two experiments were performed. In the first one, the resistance of the vector itself to mucosal environment was tested in mice. Cucumber mosaic virus particles fed to mice were able to elicit specific mucosal and serum antibody production. In the second experiment, recombinant cucumber mosaic virus fed to piglets resulted in the appearance of porcine circovirus specific serum antibodies. The vector proved to be able to survive in the gastrointestinal tract, so that an epitope expressed on its surface could induce specific immune response. These results indicate that the developed plant virus based expression system offers an effective method for mucosal vaccine production.

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Acta Veterinaria Hungarica
Authors: Márta Lőrincz, Imre Biksi, Simon Andersson, Attila Cságola, and Tamás Tuboly

Transmissible gastroenteritis (TGE) is a coronavirus-induced disease of pigs, characterised by diarrhoea and vomiting. The incidence of the disease had been decreasing since the late 1980s when deletion mutant variants (porcine respiratory coronavirus, PRCoV) of the virus emerged, repressing TGE gradually. Although disease manifestations are infrequent, the virus is still present in pig herds, causing sporadic outbreaks in a milder form. Identification and characterisation of the spike genes from TGEV and PRCoV, detected in such outbreaks, were performed in Hungary. Analysis of the amplified partial gene sequences showed that TGEV was present in herds with TGE clinical signs together with PRCoV. The sequences, apart from the deletions in PRCoV, were identical and at least two types of PRCoV spike proteins could be identified based on the length of the deleted sequence.

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Acta Veterinaria Hungarica
Authors: Márta Lőrincz, Attila Cságola, Imre Biksi, Levente Szeredi, Ádám Dán, and Tamás Tuboly

Porcine circoviruses (PCV) are present worldwide, infecting domestic pigs and wild boars alike. Studies under laboratory conditions indicated that PCV can be taken up by mice and the virus can replicate in these animals. The possible role of rodents in maintaining and transmitting PCV2 infection in the field has not been investigated yet. The present study reports the detection of PCV2, the pathogenic form of the virus, in mice and rats. A number of rodents, such as mice, rats and voles, were collected at PCV2-infected farms and also outside pig herds and tested for the presence of the virus by polymerase chain reaction (PCR). The results indicated that PCV2 can be present both in mice and rats (65.0% and 23.8% positivity, respectively) on the infected premises, but those rodents that were collected outside pig farms remained negative for PCV2.

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Acta Veterinaria Hungarica
Authors: Daniel Cadar, Attila Cságola, Marina Spinu, Ádám Dán, Krisztina Ursu, Márta Lőrincz, and Tamás Tuboly

Porcine circoviruses (PCV) are widespread in domestic pigs worldwide and there is growing information about the presence of PCV in other suid species. Based on serological studies with sera of wild boars, it was established that PCV1 was present in these animals and antibodies specific to PCV2 were also detected in wild boars living in captivity or in sylvatic areas, both with or without clinical signs of PMWS. Studies including PCV2 genome or antigen detection confirmed the previous findings. This is the first report about the presence of PCV in Transylvanian wild boar populations. Four hundred and sixty-nine samples were collected and grouped according to geographic origin, tested for the presence of PCV DNA using a real-time quantitative polymerase chain reaction assay, and 13.52% of the animals proved to be positive for one or in three cases both of the PCV genotypes. PCV2 was detected in all of the PCV-positive samples.

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Acta Veterinaria Hungarica
Authors: Anna Valkó, Imre Biksi, Attila Cságola, Tamás Tuboly, Krisztián Kiss, Krisztina Ursu, and Ádám Dán

Porcine epidemic diarrhoea virus (PEDV) can cause a severe enteric disease affecting pigs of all ages. In January 2016, diarrhoea with occasional vomiting was observed in a small pig farm in Hungary. All animals became affected, while mortality (of up to 30%) was only seen in piglets. Samples from different age groups and the carcass of a piglet were examined by various methods including pathology, bacteriology and molecular biology. PEDV was confirmed by PCR and its whole genome sequence was determined. The sequence PEDV HUN/5031/2016 showed high identity with recently reported European viruses. Differences were found mostly in the S gene, where recombination was detected with a newly identified and already recombinant swine enteric coronavirus (Se-CoV) from Italy. The present report describes the first porcine epidemic diarrhoea outbreak in Hungary after many years and gives an insight into the genetics of the Hungarian PEDV.

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Acta Veterinaria Hungarica
Authors: József Szabó, Emese Andrásofszky, Tamás Tuboly, András Bersényi, Andrea Weisz, Nikoletta Hetényi, and István Hullár

The objective of this study was designed to test whether supplementation of the diet with arginine (Arg) or glutamine (Gln) or their combination influences the production, organ weights and humoral immune response of broilers. A total of 432 one-day-old male Ross 308 broiler chickens were divided into 6 treatment groups: control, Arg-0.5%, Arg-1%, Gln-0.5%, Gln-1% and Arg-0.5%+Gln-0.5%. Drinking water and feed were provided ad libitum. On day 18 of the experiment 50% of chickens in each treatment group were immunised with bovine serum albumin. Ten and 21 days after immunisation blood samples were collected to determine the anti-albumin IgY titre, interleukin 6 (IL6) and interferon gamma (IFNG) and to measure the weight of the liver, spleen, bursa of Fabricius and thymus. Arg or Gln supplementation of the diets influenced neither the production nor the organ weights until 18 days of age. Between 18 and 39 days of age both Arg (0.5% and 1%) and Arg + Gln supplementation improved the feed conversion ratio (FCR) by 3.7%, 6.3% and 4.9%, respectively, while Gln-1% worsened it by 15%. Immunisation slightly (−0.79%) depressed the body weight gain of broilers fed the control diet, which was significantly improved by both Arg (0.5 or 1%) and Arg + Gln supplementation. Immunisation increased the weight of the spleen, bursa and thymus and decreased that of the liver. Supplementation with 1% Gln depressed (−5.13%) the body weight gain of the immunised chickens but strongly stimulated the immune response. Supplementations with Arg and Gln did not influence the IL6 and IFNG level of the blood; however, on day 10 after immunisation these two parameters showed a negative correlation with each other. Regarding production, organ weights and immunity, Arg supplementation should be recommended in the grower phase, while Gln supplementation can be useful in pullets raised for egg production, where a good immune response to vaccinations is an important factor.

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Acta Veterinaria Hungarica
Authors: József Szabó, Gergely Maróti, Norbert Solymosi, Emese Andrásofszky, Tamás Tuboly, András Bersényi, Geza Bruckner, and István Hullár

Abstract

The purpose of this 30-day feeding study was to elucidate the changes, correlations, and mechanisms caused by the replacement of the starch content of the AIN-93G diet (St) with glucose (G), fructose (F) or lard (L) in body and organ weights, metabolic changes and caecal microbiota composition in rats (Wistar, SPF). The body weight gain of rats on the F diet was 12% less (P = 0.12) than in the St group. Rats on the L diet consumed 18.6% less feed, 31% more energy and gained 58.4% more than the animals on the St diet, indicating that, in addition to higher energy intake, better feed utilisation is a key factor in the obesogenic effect of diets of high nutrient and energy density. The G, F and L diets significantly increased the lipid content of the liver (St: 7.01 ± 1.48; G: 14.53 ± 8.77; F: 16.73 ± 8.77; L: 19.86 ± 4.92% of DM), suggesting that lipid accumulation in the liver is not a fructose-specific process. Relative to the St control, specific glucose effects were the decreasing serum glucagon (–41%) concentrations and glucagon/leptin ratio and the increasing serum leptin concentrations (+26%); specific fructose effects were the increased weights of the kidney, spleen, epididymal fat and the decreased weight of retroperitoneal fat and the lower immune response, as well as the increased insulin (+26%), glucagon (+26%) and decreased leptin (–25%) levels. This suggests a mild insulin resistance and catabolic metabolism in F rats. Specific lard effects were the decreased insulin (–9.14%) and increased glucagon (+40.44%) and leptin (+44.92%) levels. Relative to St, all diets increased the operational taxonomic units of the phylum Bacteroidetes. G and L decreased, while F increased the proportion of Firmicutes. F and L diets decreased the proportions of Actinobacteria, Proteobacteria and Verrucomicrobia. Correlation and centrality analyses were conducted to ascertain the positive and negative correlations and relative weights of the 32 parameters studied in the metabolic network. These correlations and the underlying potential mechanisms are discussed.

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