In Romania, exposure to Toxoplasma gondii occurs twice as often in swine raised for familial consumption as in hunted wild boar, but occurs rarely, if ever, among fattening pigs raised in confinement.

A wide range of swine husbandry practices prevail in Romania: pork for human consumption is derived from hunting wild boar, from household rearing of small numbers of backyard pigs, and from indoor, industrial production of swine raised in confinement indoors. Romania thus represents an instructive place for evaluating the influence of animal management on the exposure risk of the zoonotic parasite, Toxoplasma gondii. The fact that many Romanians eat uncooked or undercooked pork, especially when raised for household consumption, elevates the public health imperative to understand these risks. The aim of the study, therefore, was to evaluate the seroprevalence of T. gondii in pigs and wild boars from Romania. During 2008-2010, we collected 3,595 serum samples from pigs (fattening pigs, sows, backyard pigs) and 150 serum samples from wild boars. The sera were assayed by immunofluorescence antibody test (cutoff, 1:32) for antibodies against T. gondii. The overall seroprevalence of T. gondii infection was 23.1 % (829/3,595) in pigs and 16 % (24/150) in wild boars. The seroprevalence was significantly higher (p < 0.001) in backyard pigs (30.5 %; 783/2,564) than in sows (12.4 %; 46/371) or fattening pigs (none of the sera was positive). The management system (indoor pigs versus backyard pigs) represented the most important factor in the epidemiology of T. gondii infection. The proximity of backyard pigs to the definitive host of this parasite (cats), as well as their access to contaminated meat products, elevated their exposure risk well above that of pigs raised in confinement, and even above that of wild boars inhabiting sylvatic environments.

Molecular Survey of Metastrongyloid Lungworms in Domestic Cats (Felis silvestris catus) from Romania: A Retrospective Study (2008-2011).

: Background: Lungworms are recognized as important agents in the pathology of the respiratory system in domestic cats. While Aelurostrongylus abstrusus is worldwide known and studied, Troglostrongylus brevior has gained the attention of the scientific community only in the last decade. The pathogenicity of this species seems to be higher than A. abstrusus, causing more severe clinical presentations and being potentially fatal, especially in young animals. METHODS: In this study, 371 DNA isolates of faecal samples were tested by multiplex polymerase chain reaction for the presence of A. abstrusus, T. brevior, and Angiostrongylus chabaudi. RESULTS: The results showed that 30.2% and 6.7% of the investigated domestic cats were positive for A. abstrusus and T. brevior respectively, stressing out the importance of these parasites as agents of respiratory conditions in domestic cats from Romania. None of the samples were positive for A. chabaudi. The age, the outdoor access, and the lack of deworming were identified as significant risk factors for infection with A. abstrusus. CONCLUSIONS: This paper represents the first report of T. brevior in domestic cats from Romania. Moreover, it presents the most northern localization in Europe of T. brevior.

Prevalence and genetic characterization of Toxoplasma gondii in naturally infected backyard pigs intended for familial consumption in Romania.

BACKGROUND: Foodborne toxoplasmosis in humans can be due to the exposure to tissue cysts of Toxoplasma gondii through the consumption of meat, including pork, of infected animals. Traditional Romanian food habits include pork as the preferred meat, while backyard pig rearing remains a common practice in many rural areas of Romania. The aims of the present study were to estimate the prevalence of T. gondii infection in naturally infected backyard pigs slaughtered for familial consumption and to genetically characterize the T. gondii strains obtained. METHODS: Paired blood and heart samples were collected from 94 backyard pigs, home slaughtered for private consumption. Serum samples were analyzed using the immunofluorescence antibody test (IFAT) for anti-T. gondii antibody detection. Heart samples were screened by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529) for T. gondii detection. In addition, heart samples from IFAT positive animals were bioassayed in mice. The T. gondii isolates were genotyped by the analysis of 15 microsatellite markers. RESULTS: The results showed that almost half of the pigs investigated were T. gondii seropositive (46.8%, 95% confidence interval (CI): 36.4-57.4%) and in more than a quarter of the pigs (26.6%, 95% CI: 18.0-36.7%), the parasite was detected by PCR. Three (3/44) T. gondii strains were isolated from hearts of seropositive pigs and they all belonged to genotype II. CONCLUSIONS: The present study showed the presence of T. gondii infection in backyard pigs in Romania, which suggests that consumption of pork from animals reared and slaughtered at home may pose a potential threat to human health and should be given attention. In addition, to our knowledge, this is the first study to provide data concerning T. gondii strains circulating in pigs from Romania.

Wild Birds in Romania Are More Exposed to West Nile Virus Than to Newcastle Disease Virus.

The aim of this study was to evaluate the seroprevalence of West Nile virus (WNV) and Newcastle disease virus (NDV) in wild and domestic birds from Romania. During 2011-2014, 159 plasma samples from wild birds assigned to 11 orders, 27 families, and 61 species and from 21 domestic birds (Gallus gallus domesticus, Anas platyrhynchos domesticus) were collected. The sera were assayed by two commercial competitive enzyme-linked immunosorbent assay (cELISA) kits for antibodies against WNV and NDV. We found a high prevalence of WNV antibodies in both domestic (19.1%) and wild (32.1%) birds captured after the human epidemic in 2010. Moreover, the presence of anti-NDV antibodies among wild birds from Romania (5.4%) was confirmed serologically for the first time, as far as we are aware. Our findings provide evidence that wild birds, especially resident ones are involved in local West Nile and Newcastle disease enzootic and epizootic cycles. These may allow virus maintenance and spread and also enhance the chance of new outbreaks.

Toxoplasma gondii in horse meat intended for human consumption in Romania.

The prevalence of Toxoplasma gondii, an economically important zoonotic protozoan, was investigated in horses slaughtered for export and human consumption in the North of Romania. Pairs of samples, sera and heart tissues, were collected from 82 slaughtered horses. Examination of horse sera by ELISA at a dilution of 1:10, and by modified agglutination test (MAT) at a dilution of 1:6, revealed that 32 (39%) and 31(37.8%) horses, respectively, had antibodies against T. gondii. Using polymerase chain reaction (PCR) analysis, T. gondii DNA was not found in any heart sample collected from horses. By bioassay in mice, we obtained viable isolates of T. gondii from two of ten horses determined to be strongly positive by serological assay/ELISA. The prevalence estimated in horses highlighted the potential risk for human contamination by consumption of raw or undercooked meat.

Red foxes (Vulpes vulpes) in Romania are carriers of Toxoplasma gondii but not Neospora caninum.

Brain samples from 182 red foxes (Vulpes vulpes) from Romania were examined using a standard PCR technique. Results provide evidence of Toxoplasma gondii (11 foxes=6.0%) and Neospora caninum (1 fox=0.5%) DNA in red foxes from Romania. No coinfections were found.

First isolation and genetic characterization of a Toxoplasma gondii strain from a symptomatic human case of congenital toxoplasmosis in Romania.

Very limited data exists on the genetic diversity of Toxoplasma gondii from Eastern Europe. We present the first Romanian case of symptomatic congenital toxoplasmosis in which the T. gondii strain was isolated after inoculation in mice of a cerebrospinal fluid sample from a living neonate. The T. gondii strain was genotyped with 15 microsatellite markers distributed on 10 of the 14 chromosomes of T. gondii. The strain had a type II genotype.

Northern white-breasted hedgehogs Erinaceus roumanicus as hosts for ticks infected with Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in Romania.

Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are two tick-borne pathogens of medical and/or veterinary importance which are distributed worldwide. Erinaceus roumanicus, the Northern white-breasted hedgehog, is a common synanthropic species that is known to carry not only the hedgehog tick, Ixodes hexagonus, but also I. ricinus, the most common European tick species. I. ricinus is the main vector of both mentioned pathogens. Within this framework and because only limited information is available on the role of E. roumanicus in the ecology of B. burgdorferi s.l. and Anaplasma phagocytophilum in Europe, we carried out an epidemiological surveillance on this species in Romania. From the 57 examined hedgehogs collected in 12 different counties, 24 presented tick infestation. Most ticks (n=959) were morphologically identified as larvae, nymphs, or adults of I. ricinus. The prevalence of B. burgdorferi s.l. was 0.4%, and that of A. phagocytophilum 12%. In all positive cases for B. burgdorferi s.l., restriction fragment length polymorphism revealed the genospecies B. afzelii. In Romania, only limited information is available on the epidemiology of B. burgdorferi s.l. and A. phagocytophilum. As hedgehogs commonly share the same environment with humans and other potential reservoir hosts for tick-borne pathogens, our study provides new epidemiological data of public health importance.