First report of <Emphasis Type="Italic">Trichinella britovi</Emphasis> in Serbia

In Europe, Serbia ranks among countries with a high prevalence of Trichinella infection in pigs, which continues to be a serious human health problem. While in some Balkan countries, more than one Trichinella species/genotype has been described in both the sylvatic and domestic cycles, these data are lacking for Serbia. To date, only a few Serbian isolates of Trichinella have been genetically specified, and all were classified as T. spiralis. Although transmission of Trichinella from domestic pigs to wildlife could be assumed, neither the infection status nor the species of Trichinella circulating among wildlife in Serbia has been investigated. This study shows the presence of two Trichinella species, T. spiralis and T. britovi, in wild animals originating from five districts in Serbia, where Trichinella infections in domestic pigs and humans have been recorded. Trichinella spiralis was detected in jackals (n = 3), red foxes (n = 2) and a wild cat (n = 1). We also established that wolves (n = 4) and red foxes (n = 2) serve as sylvatic reservoirs for T. britovi. This is the first report on the presence of T. britovi in Serbia.     

Epidemiology, histpathology, and muscle distribution of Trichinella T9 in feral raccoons (Procyon lotor) and wildlife of Japan

The prevalences of Trichinella T9 in trapped raccoons (Procyon lotor) and several other potential mammalian reservoirs in Hokkaido, Wakayama, and Nagasaki Prefectures were investigated. Muscle samples were collected from 2003 to 2006 from 1,080 raccoons, 113 raccoon dogs including 2 species (Nyctereutes procyonoides albus and N. p. viverrinus), 41 wild boars (Sus scrofa leucomystax), 14 martens (Martes melampus), 10 badgers (Meles meles), 5 Siberian weasels (Martes sibirica coreana), 7 mink (Mustela vison), and 1 red fox (Vulpes vulpes japonica). The samples were digested, and the prevalence and mean intensity of infection with the Trichinella muscle larvae were determined. The prevalence and intensity of the muscle larvae were 0.9% and 93.3 larvae/g (range 0.4–201.8) in raccoons, and 1.6% and 61.6 larvae/g in raccoon dogs, respectively. The infected animals were captured in different areas in Hokkaido Prefecture. These results confirmed that raccoons, which have been introduced from North America since the 1970s, are involved in the sylvatic cycle of Trichinella in Japan. In raccoons, the muscle density of Trichinella T9 larvae was highest in the tongue, and larvae were not found in the heart muscle or diaphragm. This is the first report of Trichinella T9 infection of feral raccoons in Japan.     

Apicomplexan parasites of red foxes (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) in northeastern Poland

Molecular detection of apicomplexan parasites in splenic samples of red foxes collected from northeastern Poland was conducted by PCR amplification of a fragment of the 18S rRNA spanning the V4 gene region of Apicomplexa. Positive PCR products were further analysed by restriction fragment length polymorphism (RFLP) and sequencing to identify species. One hundred and eleven red foxes (Vulpes vulpes) were acquired from 15 localities in the Mazovian Province and 27 foxes were acquired from the Mazurian Lakeland. Apicomplexan 18S rDNA was detected in 15.9% of 138 fox spleens examined. Three apicomplexan species were identified: Hepatozoon canis was detected in 11.6% of the spleen samples, Toxoplasma gondii was detected in 3.6% of the spleen samples and a Babesia sp. was sequenced from 1 sample (0.7%). This data represent the first record of H. canis, T. gondii and a B. sp. from naturally infected red foxes in Poland. Infected foxes may act as sylvatic reservoirs of these apicomplexan parasites as well as serving as a source of infection for arthropod definitive hosts and vectors.     

Detection of<Emphasis Type="Italic"> Echinococcus multilocularis</Emphasis> antigens in faeces by ELISA

Faecal samples deriving from 391 animals belonging to nine species (polecats, badgers, martens, weasels, rats, dogs, cats, red foxes, raccoon-dogs) were examined by capture ELISA for the presence of the Echinococcus multilocularis coproantigen. The main claim of our studies is the reliable detection of E. multilocularis coproantigens, mainly in the faeces of foxes, dogs and cats. For the first time in coproantigen detection we used a double-sandwich ELISA. The main advantage of this method is the higher specificity and better differentiation of positive and negative faecal samples, in comparison with sandwich ELISA. The overall specificity of double-sandwich ELISA was 95.1% with only 16 of 327 E. multilocularis-free animals giving false-positive results. The E. multilocularis coproantigen was detected by double-sandwich ELISA in 37.5% of examined red foxes and in 8.0% of examined raccoon-dogs, compared with a prevalence of just 29.8% in red foxes and 8.0% in raccoon-dogs, as determined by parasitological techniques.     

Red fox (Vulpes vulpes) cannibalistic behaviour and the prevalence of Trichinella britovi in NW Italian Alps

Food habits of the red fox (Vulpes vulpes) were studied in the Aosta Valley region (NW Italian Alps) and were related to the prevalence of Trichinella infection in the red fox itself and in two Mustelid species (the stone marten (Martes foina) and the badger (Meles meles)). The search of Trichinella by the automatic digestion of muscles samples led us to determine a prevalence of 3.5±1.2% in red foxes, 7.9±4.3% in stone martens and 1.9±1.8% in badgers, with no significant differences among the species. All larvae were identified as Trichinella britovi. The fox diet was assessed through the analysis of both faeces (n=180) and the stomach contents of road-killed animals (n=109). Our results confirmed the opportunistic feeding behaviour of the red fox, which is able to use various trophic resources, both of animal and vegetal origin: e.g. wild and cultivated fruits (F%=47.1; V%=67.3), rodents (F%=22.8; V%=64.8) and carrion (F%=15.6; V%=78.6) formed the bulk of the foxs diet. The frequency of occurrence of potential events of cannibalism was 1.0%, even if the complete absence of undigested remains, other than hairs, suggested the possibility of confusing cannibalism with coat-cleaning. We suggest that intra-specific necrophagy could not represent the unique way of transmission of the nematode in natural conditions.     

Survey of Toxoplasma gondii and Trichinella spp. in hedgehogs living in proximity to urban areas in the Czech Republic

Hedgehogs (Mammalia: Erinaceidae) are omnivorous nocturnal animals typically living in anthropogenic areas. They may be suitable as sentinels for a wide range of zoonotic infections. Only a few studies have investigated hedgehogs (and then as representative wildlife species) to establish their role in the life cycle of such tissue parasites with zoonotic potential as Toxoplasma gondii or Trichinella spp. Working with frozen hedgehog cadavers, we tested for these parasites using T. gondii DNA-specific magnetic capture isolation plus polymerase chain reaction and Trichinella spp. digestion assay. All of 50 examined hedgehogs were negative for Trichinella spp. larvae in their muscles, but brain tissue from 5 out of 26 Erinaceus europaeus (19.2%) and 4 out of 24 E. roumanicus (16.6%) tested positive for T. gondii DNA. Frequency of T. gondii for both hedgehog species was equal, as was distribution between males and females and across age categories. Although a few studies have suggested the possibility of Trichinella spp. infection in hedgehogs, the zero prevalence in the tested hedgehogs is not surprising in view of the generally low prevalence of Trichinella spp. in Central Europe. Our results show that hedgehogs are susceptible to infection by T. gondii and can be used as indicator wildlife animal species in anthropogenic ecosystems.

     

Molecular identification of Trichinella britovi in martens (Martes martes) and badgers (Meles meles); new host records in Poland

Trichinella larvae were detected in a marten (Martes martes) and a badger (Meles meles) in Poland. The animals were found dead following car accidents. All examined animals derived from the Mazurian Lake district, north-east Poland, near the village Kosewo Górne where Trichinella infection were earlier confirmed in wildlife; red foxes and wild boars. The muscle samples were examined by artificial pepsin-HCl digestion method. The parasites were identified as Trichinella britovi by multiplex polymerase chain reaction method. Larvae were found in two out of three martens and one out of seven examined badgers. This is the first report of the identification of Trichinella britovi larvae from martens and badgers in Poland.     

Occurrence of Echinococcus multilocularis in red foxes from the Carpathian regions of Slovakia and Poland

The extensive distribution of Echinococcus multilocularis cestode from endemic alpine areas to the parts of Central Europe has been recorded in recent years. The first confirmed finding of E. multilocularis in Slovakia was recorded in 1999 in the area adjacent to the Polish border. At present, this serious zoonosis occurs almost across the whole territory of Slovakia. The occurrence of these tapeworms in red foxes (Vulpes vulpes) at the border regions of Slovakia and Poland has been monitored. In these districts, out of 152 faecal samples examined, 36.2% were positive for the coproantigen-ELISA. With the sedimentation and counting technique the prevalence of E. multilocularis in red foxes was up to 38.8%. The examination of foxes from neighbouring districts revealed worm burden ranging from 1–15,000 specimens, but the majority of animals harboured medium number of tapeworms. In the Small Carpathian and Sub-Carpathian regions of Poland, out of 65 samples examined, 13.8% were coproantigen positive. Using the small intestine scraping method only 6.1% prevalence of E. multilocularis in red foxes was determined, mostly with a high worm burdens over 1,000 specimens. The results suggest possible transborder transmission of E. multilocularis, the causative agent of serious alveolar echinococcosis.     

Trichinella infection in wild animals from endemic regions of Argentina

Natural infection with Trichinella has been described in more than 150 mammalian species. However, few reports of Trichinella infection in wild animals have come from Argentina. In this study, muscle tissue was obtained from wild animals in Argentina with the aim of evaluating the presence of Trichinella. A total of 169 muscle samples were collected to determine the presence of Trichinella larvae by artificial digestion. The 169 muscle samples originated from 12 species including 36 opossums (Didelphis albiventris), 19 armadillos (Chaetophractus villosus), 9 capybaras (Hydrocaeris hydrocaeris), 1 puma (Puma concolor), 3 grey fox (Lycalopex gymnocercus), 6 coypus (Myocastor coypus), 6 skunks (Conepatus chinga), 2 ferrets (Galictis cuja), 66 rats (Rattus norvegicus), 6 mice (Mus musculus), 12 wild boars (Sus scrofa), and 3 wild cats (Felis geoffroyi). Trichinella infection was detected in 1 puma [2 larvae per gram (LPG)], 3 wild boars (8–420 LPG), 3 armadillos (0.04–0.08 LPG), and 9 rats (0.1 to 150 LPG). Only 3 Trichinella isolates, of 1 rat and 2 wild boars from Neuquén, were identified as Trichinella spiralis by nested PCR. The presence of Trichinella infection among wild animal populations suggests a sylvatic cycle of transmission in Argentina, which can serve as a reservoir for humans and domestic animals. Further, evidence of high prevalence in rats emphasizes the need to improve pig management, mainly in small individual farms without adequate technology, to enhance the quality of feeds, and to improve veterinary services to avoid exposure of pigs to Trichinella.     

Occurrence of <Emphasis Type="BoldItalic">Cryptosporidium</Emphasis> spp. in red foxes and brown bear in the Slovak Republic

Wild animals can be involved in epidemiology of many important diseases and often act as reservoirs of pathogens which cause disease in domestic animals and humans. This paper aims the role of red fox (Vulpes vulpes) and brown bear (Ursus arctos) in the circulation of coccidian parasites from the genus Cryptosporidium. Cryptosporidiosis is known as an important enteric pathogen, clinical symptoms in particular in immune-compromised individuals range from mild to severe diarrhoea and dehydration, which could be fatal. Fecal samples from 62 red foxes shot during September 2010 to February 2011 and 63 brown bears collected during June 2010 to March 2011 in central and eastern Slovakia were examined for the qualitative determination of Cryptosporidium spp. antigens in faeces by sandwich ELISA kit. Overall, 38.7% (24/62) of faecal samples of red foxes and 55.6% (35/63) of faecal samples of brown bear were positive. Our preliminary results emphasize prevalence of Cryptosporidium spp. amongst brown bears and red foxes in Slovakia and highlight the potential risk for transmission of cryptosporidiosis to humans using the countryside for professional or recreational purposes.     

Molecular epidemiology of <Emphasis Type="Italic">Trichinella</Emphasis> spp. in three Baltic countries: Lithuania,Latvia, and Estonia

Meat of domestic pigs and wild boars has been the significant source of emerged human trichinellosis in Lithuania, Latvia, and Estonia over the past two decades. However, there is very little known on the occurrence of Trichinella spp. in main wildlife reservoirs and its transmission in domestic and sylvatic cycles in these countries. The present study demonstrated considerably higher endemicity of Trichinella spp. in main sylvatic reservoirs (28.9–42% in foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in all three countries than previously reported. Molecular identification of Trichinella larvae from more than 500 sylvatic and domestic animals revealed four Trichinella species (Trichinella spiralis, Trichinella britovi, Trichinella nativa, and Trichinella pseudospiralis) sympatric in a relatively small area and several as the first records for the respective countries. The nonencapsulating T. pseudospiralis is found for the first time in the Eastern Europe. Sylvatic T. britovi was found in domestic pigs in Lithuania and Latvia (16 and 57.1%, respectively) and only in these countries, domestic T. spiralis was detected in sylvatic animals in areas where domestic trichinellosis was registered. The study suggests that transmission of Trichinella between domestic and sylvatic cycles in Lithuania and Latvia is favored by improper human behavior, e.g., pig and slaughter waste management.     

Helminth fauna of the red fox (Vulpes vulpes Linnaeus, 1758) in southern Belarus

We examined 94 carcasses of and 1,213 faecal samples from red foxes collected between 1981 and 2001 in natural and transformed ecosystems in the southern part of Belarus. A total of 32 helminth species were found. Of these, the most common were Alaria alata, Pearsonema plica, Taenia crassiceps, Toxocara canis, Trichinella spp. larvae and Uncinaria stenocephala. All species are significant for medical and veterinary health.     

Liver fluke (Opisthorchiidae) findings in red foxes (Vulpes vulpes) in the eastern part of the Federal State Brandenburg, Germany – a contribution to the epidemiology of opisthorchiidosis

Parasitological examination of 677 livers from red foxes shot in connection with a rabies control programme were carried out in the eastern districts of the Federal State Brandenburg/Germany in 1996. Of the foxes, 32.5% were positive for opisthorchiid flukes. Metorchis bilis, the most frequently occurring fluke was found in 28.1% of foxes with numbers between 1 and 185. Opisthorchis felineus was found in 6.7% of fox livers. A maximum of 169 specimens was removed from one animal. Pseudamphistomum truncatum was present in only 2 foxes. The percentage of positive foxes in different districts varied between 15.8 and 43.3%. Significant differences in fluke prevalence were found between districts with a high share of surface water compared with a district with a low percentage of surface water. No sex-dependent differences in opisthorchiid prevalence were established. Received: 20 May 1998 / Accepted: 18 June 1998     

Echinococcus multilocularis in south-eastern Europe (Romania)

Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE) in humans, has been found in 4.8% of 561 red foxes originating from various regions of Romania. Infected foxes were identified in 8 of 15 counties with average prevalence rates between 1.7% and 14.6%. In previous studies, E. multilocularis was not found in 535 foxes from three counties, but larval stages (metacestodes) were present in four species of rodents. Furthermore, AE was diagnosed in two patients. Experiences from other European regions indicate that several factors (such as increasing fox populations with higher parasite prevalences and urban cycles of the parasite) may result in an increased infection risk for humans.     

First report of Trichinella pseudospiralis in Poland, in red foxes (Vulpes vulpes)

Nematode worms of the genus Trichinella are one of the most widespread zoonotic pathogens. Natural transmission between hosts can only occur through the ingestion of infected meat. To date, two Trichinella species are known to be etiological agents of disease among domestic animals and wildlife in Poland: T. spiralis and T. britovi. In the last decades, since the administration of an oral vaccination against rabies, the red fox population in Poland has increased exponentially. The study area covers the Nowy Targ region: a mountainous area (585–1138 m above the sea) in southern Poland. Of 24 red foxes examined in the study, four were infected with Trichinella isolates: three were identified as T. britovi and one as T. pseudospiralis. The muscle of red foxes infected with T. britovi harboured 2.75, 3.11, 4.4 LPG and with T. pseudospiralis 0.36 LPG. Trichinella larvae were identified at species level by genomic and mitochondrial multiplex PCR, the products of which were sequenced for comparison with other sequences available in GenBank. The sequences obtained from the Polish T. pseudospiralis isolate, deposited in GenBank under the accession numbers JQ809660.1 and JQ809661.1, matched sequences already published in GenBank. Sequence comparison showed a 100% match with the large subunit ribosomal RNA gene of T. pseudospiralis isolate ISS 013, and a 96–95% match with those of T. pseudospiralis isolates ISS 141 and ISS 470. This is the first report of the identification of T. pseudospiralis larvae from red fox in Poland.     

Influence of urbanization on the epidemiology of intestinal helminths of the red fox (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) in Geneva,Switzerland

Dixenous helminths that depend on rodent intermediate hosts are supposed to be negatively affected by urbanization due to lower supply of rodents in urbanized environments. Prevalence rates of dixenous, non-strictly monoxenous, and monoxenous helminths in 228 red foxes (Vulpes vulpes) along a gradient of increasing urbanization were assessed by morphological parasite identification in the city of Geneva, Switzerland. Multivariate analyses for the five most prevalent helminth species or genera revealed a significant decrease of prevalence rates for the dixenous helminths Echinococcus multilocularis and Taenia spp. from the rural (52.1 and 54.3%, respectively) to the urban area (30.0 and 20.0%, respectively), but not for the monoxenous nematode Uncinaria stenocephala (overall prevalence of 78.2%) and the non-strictly monoxenous nematode Toxocara canis (overall prevalence of 44.3%). The lower prevalence of Toxascaris leonina in the urban area (8.0%) compared to the rural area (59.6%) raises the question of whether rodent paratenic hosts play a major role for the population dynamics of this species.     

Infections with cardiopulmonary and intestinal helminths and sarcoptic mange in red foxes from two different localities in Denmark

Monitoring parasitic infections in the red fox is essential for obtaining baseline knowledge on the spread of diseases of veterinary and medical importance. In this study, screening for cardiopulmonary and intestinal helminths and sarcoptic mange (Sarcoptes scabiei) was done on 118 foxes originating from two distinct localities in Denmark, (Copenhagen) greater area and southern Jutland. Fifteen parasite species were recorded in 116 foxes (98.3%), nine parasitic species are of zoonotic potential. Parasite diversity was greater in foxes of Copenhagen in terms of overall parasite species richness and species richness of all helminth groups individually: trematodes; cestodes; and nematodes. Six parasite species were recovered from foxes of Copenhagen, but not from foxes of Southern Jutland: Echinochasmus perfoliatus; Echinostoma sp.; Pseudamphistomum truncatum; Dipylidium caninum; Angiostrongylus vasorum; and Sarcoptes scabiei, but Toxascaris leonina was only recorded in foxes of southern Jutland. A high prevalence and abundance of A. vasorum in foxes of Copenhagen was observed. The prevalence of four nematode species; Eucoleus (Capillaria) aerophilus, Uncinaria stenocephala, Toxocara canis, and Crenosoma vulpis, in foxes of both localities were comparable and ranging from 22.9% to 89%. The prevalence of Mesocestoides sp. was significantly higher in foxes of Copenhagen. Taenia spp. were detected using morphological and molecular analysis, which revealed the dominance of T. polyacantha in foxes of both localities. Infections with sarcoptic mange were evident only among foxes of Copenhagen (44.9%), which significantly affected the average weight of the infected animals. Further remarks on the zoonotic and veterinary implications of the parasites recovered are given.     

Validation of a Western Blot for the detection of anti-<Emphasis Type="Italic">Trichinella</Emphasis> spp. antibodies in domestic pigs

Trichinellosis is a zoonotic disease in humans caused by Trichinella spp. According to international regulations and guidelines, serological surveillance can be used to demonstrate the absence of Trichinella spp. in a defined domestic pig population. Most enzyme-linked immunosorbent assay (ELISA) tests presently available do not yield 100% specificity, and therefore, a complementary test is needed to confirm the diagnosis of any initial ELISA seropositivity. The goal of the present study was to evaluate the sensitivity and specificity of a Western Blot assay based on somatic Trichinella spiralis muscle stage (L1) antigen using Bayesian modeling techniques. A total of 295 meat juice and serum samples from pigs negative for Trichinella larvae by artificial digestion, including 74 potentially cross-reactive sera of pigs with other nematode infections, and 93 meat juice samples from pigs infected with Trichinella larvae were included in the study. The diagnostic sensitivity and specificity of the Western Blot were ranged from 95.8% to 96.0% and from 99.5% to 99.6%, respectively. A sensitivity analysis showed that the model outcomes were hardly influenced by changes in the prior distributions, providing a high confidence in the outcomes of the models. This validation study demonstrated that the Western Blot is a suitable method to confirm samples that reacted positively in an initial ELISA. Frey and Schuppers contributed equally to this work. This work was funded by the Swiss Federal Veterinary Office, grant number 1.06.03.     

Predominance of Blastocystis sp. subtype 4 in rural communities, Nepal

Blastocystis sp. is a common intestinal parasite. To date, there have been sporadic and scanty studies on Blastocystis sp. carried out in rural communities in Nepal. We surveyed the prevalence of Blastocystis sp. and its possible associated risk factors, and reported the predominant Blastocystis sp. subtype in two rural communities, Bolde Phediche and Bahunipati, in Nepal. Human faecal samples were collected from 241 participants, cultured using in vitro cultivation and examined for Blastocystis sp. The presence of Blastocystis sp. in faecal samples was further confirmed by polymerase chain reaction (PCR) and subsequently genotyped using subtype-specific sequence tagged site (STS) primers. There were 26.1% (63/241) of the participants that were infected by Blastocystis sp. We detected 84.1% (53/63) of Blastocystis sp. subtype 4 infections in these rural communities. The unusually high prevalence of Blastocystis sp. subtype 4 can be attributed to the rearing of family-owned animals in barns built close to their houses. Eighty one percent (51/63) of the Blastocystis sp. infected participants drank not boiled or unfiltered water. The present study revealed that Blastocystis sp. could pose a health concern to the communities and travellers to the hilly area in Nepal. Infection may be transmitted through human-to-human, zoonotic and waterborne transmissions. We provide recommendations to ensure good public health practices.     

The prevalence of Echinococcus multilocularis in red foxes in Poland—current results (2009–2013)

The aim of the study was to determine the prevalence of Echinococcus multilocularis in red foxes (Vulpes vulpes) in Poland. Overall, 1,546 intestinal samples from 15 of the 16 provinces in Poland were examined by the sedimentation and counting technique (SCT). The mean prevalence of E. multilocularis in Poland was 16.5 % and was found in 14 of the 15 examined provinces. The mean intensity of infection was 2,807 tapeworms per intestine. Distinct differences in prevalence were observed between regions. In some provinces of eastern and southern Poland, the level of prevalence was 50.0 % (Warmińsko-Mazurskie), 47.2 % (Podkarpackie), 30.4 % (Podlaskie) and 28.6 % (Ma?opolskie), while in other provinces (west and south-west), only a few percent was found: 2.0 % (Dolno?l?skie), 2.5 % (Wielkopolskie) and 0.0 % (in Opolskie). The border between areas with higher and lower prevalence seems to coincide with a north–south line running through the middle of Poland, with prevalence from 17.5 to 50.0 % in the eastern half and from 0.0 to 11.8 % in the western half. The dynamic situation observed in the prevalence of this tapeworm indicated the necessity of continuing to monitor the situation concerning E. multilocularis in red foxes in Poland.