Imre Biksi

Detection, prevalence and analysis of emerging porcine parvovirus infections

A number of newly identified porcine parvoviruses had been described during the last decade, but the presence and prevalence of these viruses are unknown in Hungary and only partly known for Europe. The present study was conducted to detect and measure the prevalence of these viruses, namely porcine parvovirus (PPV) 2, PPV3, PPV4, porcine bocavirus (PBoV) 1, PBoV2, PBo-likeV and the 6V and 7V parvoviruses. The prevalence of PPV1 and porcine circovirus type 2 (PCV2) was also investigated. Faecal samples, blood serum samples, organ tissues, foetuses and semen were collected from different swine herds in Hungary and tested by polymerase chain reaction methods specific for the different viruses. The results indicated that all of the examined parvoviruses were present in Hungary, hence in Europe. The prevalence was 18.1% for PCV2, 0.5 % for PPV1, 6.4% for PPV2, 9.7% for PPV3, 6.4% for PPV4, 1.5% for PBo-likeV, 4.8% for PBoV1 and PBoV2 and 1.8% for 6V and 7V. Based on the analysis of partial PPV4 and PBo-likeV sequences, these viruses showed a high degree of sequence conservation, whereas PPV3 and the majority of PPV2, PBoV1, PBoV2, 6V and 7V sequences showed higher variability. Possible sites of recombination were also identified between PBoV1 and PBoV2 genomes.

Detection of porcine circovirus in rodents — Short communication

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.

Sarcocystis-infection of cattle in Hungary

BackgroundReports on Sarcocystis-infection of cattle are outdated or lacking in many European countries, including those in the Central-Eastern part of the continent. Therefore, to assess the prevalence of Sarcocystis spp. among bovids in Hungary, a countrywide survey was initiated. In addition, fulminant deaths of four cattle, that showed clinical signs and post mortem lesions resembling acute sarcocystiosis (“Dalmeny disease”), were investigated.MethodsDuring the countrywide survey individual heart and oesophagus samples were collected at slaughterhouses from 151 beef cattle and from 15 buffalo, kept in 31 places of Hungary. Analysis for Sarcocystis spp. was carried out with conventional PCRs for the 18S rDNA gene and gel electrophoresis, followed by sequencing of 36 strongly positive samples. Mortality cases were evaluated by histological, molecular, bacteriological and virological analyses of samples from various organs.ResultsAmong slaughtered cattle the rate of Sarcocystis-infection was 66%. S. cruzi was identified as the most prevalent species in aurochs-like breed, and the zoonotic S. hominis in Hungarian grey cattle. Concerning the sudden deaths of cattle, Sarcocystis-infection could not be demonstrated in organs showing haemorrhages, but S. cruzi cysts were present in the muscles. In one case “S. sinensis” was molecularly identified in the blood (indicating sarcocystaemia). Results of analyses for bacterial/viral pathogens were negative.ConclusionsS. cruzi appears to be the most prevalent Sarcocystis sp. in cattle in Hungary, followed by the zoonotic S. hominis. However, the rate of infection with both species was shown to differ between cattle breeds. The suspected role of Sarcocystis spp. as causative agents of the fatal cases could not be confirmed.

Genome Sequence of a Monoreassortant H1N1 Swine Influenza Virus Isolated from a Pig in Hungary

ABSTRACT The genome of a porcine H1N1 influenza A strain is reported in this study. The strain proved to be a monoreassortant strain with a typical porcine N1 gene on the genetic backbone of the pandemic H1N1 influenza A virus strain. Monitoring of descendants of the pandemic 2009 H1N1 strain is needed because of concerns that more-virulent strains may emerge in forthcoming epidemic seasons.

Porcine epidemic diarrhoea virus with a recombinant S gene detected in Hungary, 2016

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.

EMERGENCE AND CHARACTERISATION OF PANDEMIC H1N1 INFLUENZA VIRUSES IN HUNGARIAN SWINE HERDS

In 2010, two novel porcine H1N1 influenza viruses were isolated from pigs with influenza-like illness in Hungarian swine herds. Sequence and phylogenetic analysis of these strains revealed that they shared molecular features with the pandemic H1N1 influenza virus strains, which emerged globally during 2009. The PB2, HA and NA genes contained unique amino acid changes compared to the available new H1N1 influenza virus sequences of pig origin. Furthermore, the investigated strains could be separated with respect to parallel amino acid substitutions affecting the polymerase genes (PB2, PB1 and PA) and the nucleoprotein (NP) gene, supporting the proposed complementarities between these proteins, all required for the viral fitness. Molecular characterisation of two Hungarian human pandemic H1N1 isolates was also performed, so that we could compare contemporaneous strains of different host species origins. Shared molecular motifs in various genes of animal and human influenza strains suggested that the Hungarian porcine strains could have originated from humans through direct interspecies transmission. This study is among the few that support the natural human-to-pig transmission of the pandemic H1N1 influenza virus.

Toxoplasmosis in Tammar wallabies (Macropus eugenii) in the Budapest Zoo and Botanical Garden (2006-2010)

Smaller macropodid species (commonly referred to as wallabies) are extremely susceptible to toxoplasmosis: in most cases, infection with Toxoplasma gondii leads to death within a short time. Between June 2006 and July 2010, T. gondii was detected by immunohistochemical examination in six Tammar wallabies (Macropus eugenii) that died in the Budapest Zoo and Botanical Garden; in another four specimens histopathology revealed T. gondii-like organisms (which could not be differentiated from Neospora caninum solely by morphology), and in another 11 animals toxoplasmosis as the possible cause of death could not be excluded. The current zoo population of 12 Tammar wallabies was tested for T. gondii IgG antibodies by the modified agglutination test (MAT), with negative results. We suppose that most of the deaths were due to acute toxoplasmosis resulting from a recent infection.

Outbreak of Klebsiella oxytoca enterocolitis on a rabbit farm in Hungary

THE genus Klebsiella , a member of the family Enterobacteriaceae, includes ubiquitous bacteria found both in the environment and in animals (Murray and others 2003). As opportunistic pathogens they can cause pneumonia and urogenital infections in human beings, rodents, carnivores and ungulates, mastitis in ruminants, and septicaemia in a number of species (Quinn and others 1994, Boucher and Nouaille 2002). Enteritis and diarrhoea due to villous atrophy associated with Klebsiella species infection has been reported in newborn pigs (Brown and others 2008). Knowledge about the importance of Klebsiella species infections in domestic rabbits is limited, although Klebsiella pneumoniae has been reported to be associated with haemorrhagic enteritis and septicaemia in Italy and France (Boucher and Nouaille 1999, 2002, Coletti and others 2001). Klebsiella oxytoca may cause urinary tract infections in human beings and it has been described as a cause of antibiotic-associated haemorrhagic colitis and diarrhoea (Hogenauer and others 2006, Hoffmann and others 2010), as well as a cause of septicaemia in human infants (Reiss and others 2000). This short communication describes an outbreak of disease associated with K oxytoca among farmed rabbits in Hungary. In January 2010, a rabbit breeding …

Sporadic re-emergence of enzootic porcine transmissible gastroenteritis in Hungary

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.

Inclusion body rhinitis in pigs in Hungary.

INCLUSION body rhinitis in pigs, caused by porcine cytomegalovirus (PCMV), occurs sporadically worldwide (Done 1955, Harding 1958, Edington 1999). The virus belongs to the Betaherpesvirinae family, and although it is species specific, it shares homologies with the cyto megaloviruses of human beings and other animals (Edington 1999). PCMV infection remains mostly inapparent in older pigs, but infection of the dam during gestation can lead to fetal death and mummification, abortion, low viability of piglets at birth and consequential preweaning mortality (Edington and others 1988). Infection in threeto five-week-old piglets may occur under unfavourable conditions, with clinical signs of sneezing, nasal discharge and respiratory distress, and may even result in 20 to 25 per cent mortality. In growers, the immunosuppressive effect of the virus can lead to exacerbation of certain diseases in the herd. The virus is also thought to contribute to pneumonia in growers. Infection can be diagnosed by the presence of characteristic large, basophilic, intranuclear inclusion bodies in cytomegalic cells of the nasal glandular epithelium. Such inclusion bodies and mononuclear cellular infiltration can also be detected in the tubular epithelia of the kidneys, as well as the epithelia of the salivary and tear glands. Other, less frequent, lesions include interstitial pneumonia and lymphocytic perivasculitis in the brain (Kelly 1967, Edington and others 1976). Predisposing factors of the infection are not fully known. A low level of immunity within the herd (for example, in newly established herds) and immunosuppressive effects may play a role (Edington 1999). Seroconversion due to PCMV is probably much more frequent than clinical disease (Lim and others 2002). A PCR and nested PCR have recently been developed for the specific detection of PCMV nucleic acid (Hamel and others 1999, Lim and others 2002). This short communication describes inclusion body rhinitis in pigs in Hungary for the first time. From mid-December 2002, severe losses occurred among sucking piglets in a herd of 350 farrow-to-finish pigs in southern Hungary. Piglets with apparently normal vigour at birth died without clinical signs at two to three days of age; a similar problem was observed later in twoto three-weekold piglets. Total piglet mortality in the farrowing houses approached 40 per cent at its worst, with several episodes of high mortality observed between mid-December 2002 and mid-April 2003. Sow mortality was low and unaffected, and the incidence of postparturient hypogalactia syndrome and/or discharges after farrowing did not increase. Litters from primiparous and older sows were equally affected. The number of stillbirths per litter rose slightly, but the number of mummified piglets per litter was unaffected. There was a decrease in farrowing rate of approximately 10 per cent starting from mid-September 2002 and ending in March 2003. Pregnancy losses were attributed to an increase in both regular and irregular repeats and the number of sows ‘not in pig’. Almost the whole herd of breeding sows had to be replaced. Replacement breeding gilts were purchased from the original source, which did not display any clinical signs of infection at that time. Unfortunately, there was no close veterinary supervision when the problem started, so diagnostic efforts were not initiated until the end of March 2003. FIG 1: (a) Cytomegaly and large basophilic inclusion body formation in glandular epithelial cells (arrow). Haematoxylin and eosin. x 100. (b) Porcine cytomegalovirus (arrow) in glandular epithelial cells. Transmission electron microscopy. x 25,000