Open Access
Research Article
Issue
Parasite
Volume 20, 2013
Article Number 3
Number of page(s) 5
DOI https://doi.org/10.1051/parasite/2012003
Published online 14 January 2013

© V. Lalošević et al., published by EDP Sciences, 2013

Licence Creative Commons
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Introduction

Nematodes of the family Capillariidae Neveu-Lemaire, 1936 are a very large group with not yet completely accepted taxonomy [16, 32]. Among species ranked in this family, Eucoleus aerophilus (Creplin, 1839) Dujardin, 1845 (syn. Capillaria aerophila or Thominx aerophilus) affects trachea and main bronchi of canids, felids and some omnivorous animals [19]. Although several morphological descriptions have been provided for different biological stages of E. aerophilus [3, 5, 18, 23, 32], several other aspects of this nematode are still poorly known. Indeed, this is the case of geographic distribution, clinical significance and actual zoonotic potential [31]. Also, knowledge on the biological cycle of this nematode is scanty and, for instance, the route of transmission to vertebrate hosts remains to be understood. In general it is thought that animals become infected by ingesting environmental larvated eggs [31] but it has been also hypothesized, but never demonstrated, that larval E. aerophilus might require the passage through earthworms to become infective for cats and foxes (Borovkova, 1947 cited in [18]).

In the past few years some evidences have suggested that E. aerophilus is spreading in several geographic areas and that there is an increase of cross-infections between wild and domestic animals [79, 30, 31]. Indeed, it has been preliminarily shown that some E. aerophilus populations are shared between domestic pets and wildlife, e.g. red fox (Vulpes vulpes) and beech-marten (Martes foina (Erxleben) [9]).

Such changes are a likely effect of the increase of fox populations in peri-urban and urban areas and of movements of wild and companion animals around regions. In fact, an increase of the red fox population has been documented in Europe after the success of the oral vaccination against rabies [4, 33]. The role of the red fox as reservoir for parasites may be now of relevance especially in suburban areas, e.g. for multilocular echinococcosis and other parasitic diseases of veterinary (e.g. angiostrongylosis) and human (e.g. trichinellosis) concern [11, 15, 21, 22, 31].

With regard to Serbia, E. aerophilus is circulating in cats and foxes [12, 13] and, interestingly, the last case of human capillariosis documented in the literature is from the same country. More specifically, an infected woman showed respiratory symptoms miming bronchial carcinoma and blood eosinophilia [12]. Considering all the aforementioned background, especially the potential risk for human health which causes new interest in capillariid nematodes, the aim of this work was to investigate the infection rate of E. aerophilus in red foxes of Serbia.

Materials and Methods

Study area

In winter season from 2008 to the end of 2011, foxes (n = 118) hunted in the framework of a rabies control campaign were collected from the whole territory (21,506 square km) of the Vojvodina province of Serbia (45°15 N, 19°50′ E). This province is crossed by the Danube river, two tributaries (Tisza and Sava) and many canals of the Danube-Tisza-Danube system (Pannonian Basin) on the left side and the Fruska Gora mountain on the right. The climate is continental.

Sampling

All foxes were adults with a body weight ranging from 4.0 to 8.1 kg. Carcasses were opened and the tracheas of 70 foxes, from the larynx to the main bronchi bifurcation, were collected and preserved in 30% ethanol. Tracheas were then opened on the anterior side by scissors and parasites retrieved were collected under a stereomicroscope and wet-mounted in glycerine-ethanol or lactophenol. The nasal cavity of 10 foxes was opened through the palate, and the nasal mucosa was scrapped and conserved in 30% ethanol. Faecal samples were collected during the necropsy of the 118 foxes and tested by the glycerine flotation method, respectively. Parasites collected were counted, photographed by a Leica microscope and measured by the “ImageJ” free program (http://imagej.nih.gov/ij/). Adult stages of nematodes recovered from trachea and bronchi of the foxes were identified at the species level by morphological keys [17, 23]. Five adult female and three male E. aerophilus voucher specimens were deposited in the Zoological collection of the Department of Biology and Ecology, University of Novi Sad, Serbia (Registration No. NEM-0037).

Results

Adults of E. aerophilus (i.e. total of 817) were detected in 59 foxes (84%), with an average worm burden of 14 (range 1–71) (Figures 1 and 2). Out of 70 examined foxes, 32% harboured more than 14 adult worms of E. aerophilus with no relevant difference in parasitic burden between foxes collected from different mountainous regions or the Pannonian basin. The sex was assessed for 390 out of 817 examined worms because some E. aerophilus specimens were damaged during the collection. The sex ratio between male and female worms was 1:2.56.

thumbnail Figure 1.

Adult female Eucoleus aerophilus on the mucosa of the opened trachea of a fox dissected after formalin fixation. More than half of the worm body is inside the tracheal mucosa with a zig-zag shape and the other part is free in the lumen. Bar, 1 mm.

thumbnail Figure 2.

Number of Eucoleus aerophilus adults per foxes.

Morphometric measures were obtained from 82 males and 75 females. Measurements are in micrometer (μm) and indicated here as mean and range between parentheses. Body length, male 19,020 (10,410–25,110), female 27,980 (16,010–41,840); mean diameter measured after end of oesophagus, male 86 (69–110), female 147 (107–185); oesophagus length, male 6,590 (4,390–7,980), female 6,003 (4,540–7,330); number of stichocytes, calculated in 10 males and 11 females, male 44 (37–46), female 44.5 (35–50). The lateral bacillary bands were well recognized but the ventral bacillary bands were observed rarely. The uterus containing numerous eggs continued in a muscular vagina with a vulvas opening (Figure 3). Eggs were unembryonated, thick-shelled with a reticulated surface and with polar plug-like opercula, giving them a lemon shape. Eggs not symmetrical, opercular axes slightly angulated. Measurements of eggs in the final part of the uterus of 75 females: 73 × 35 (61–95 × 30–47). The caudal end of males showed a small pseudobursa with two bulges next to the cloaca opening. Males showed a single very thin spicule and a spicule sheath with many spines (Figure 4).

thumbnail Figure 3.

Female Eucoleus aerophilus showing the end part of the last stichocyte with the cell nucleus (N), two superimposed secretory cells (S), intestine (I), vagina with eggs (E) and vulva (arrowhead). Bar, 50 μm.

thumbnail Figure 4.

Male Eucoleus aerophilus, extruded spinose spicule sheath and partially extruded spicule at the caudal part. Bar, 100 μm.

For 10 foxes the nasal cavity mucosa was scrapped and examined under a stereomicroscope. Adult stages belonging to the closely related species Eucoleus boehmi (Supperer, 1953) were found in nine animals. The average worm burden was seven (range 1–20).

The results of the copromicroscopic examinations on faecal samples collected at the necropsy are reported in Table 1.

Table 1.

Results for the copromicroscopic examinations of the foxes (n = 118) examined in the present study. N: number of foxes scored positive for endoparasites.

Discussion

The infection rate of E. aerophilus infection (84%) herein detected in foxes from the Pannonian and the Fruska Gora Mountain regions (Vojvodina province) is indeed high. Interestingly, along with the results (88%) recently obtained from foxes of Norway [6], this study demonstrated the highest infection rate of pulmonary capillariosis in wild canids in Europe.

In the Zagreb region (Croatia) which shows a similar landscape to the central part of Serbia, about 300 km on the West, the prevalence of E. aerophilus in foxes was only 4.7% with a highest worm burden of five worms [20]. In Italy, the infection rate of E. aerophilus detected in the trachea of foxes was 7.0% in the Tuscany region [14]. In the Netherlands and Hungary E. aerophilus was found in 46.8% and 66% of dissected foxes [1, 27]. An infection rate by “Capillaria spp.” of 22.4% was found in foxes from Slovakia by copromicroscopic investigation [15].

Albeit the high prevalence of E. aerophilus in foxes from Serbia could be due to the high sensitivity of the diagnostic methods used, most of the aforementioned studies have relied on trachea dissection as well. A great discrepancy between copromicroscopic and necroscopical findings of capillariids in the present study demonstrates that copromicroscopy may fail in terms of diagnostic sensitivity for this infection in foxes. Therefore, factors other than the diagnostic approach should be implicated in influencing such a high infection rate by lung capillariosis in foxes from Serbia.

The high humidity of the habitat of Vojvodina, the South part of Pannonian Basin, with many canals between the Danube and Tisza rivers, and many small tributaries of the Sava river in the Fruska Gora Mountain could favour parasite’s dissemination. In this habitat, the fox density is of over 0.2 per square km and the high humidity can favour earthworms’ dispersion. Also, competitive pressure and other diseases (e.g. sarcoptic mange) can enhance the susceptibility of foxes to nematodes [24] and the increase of reports of capillariosis in companion animals suggests that E. aerophilus is spreading in several areas. Epidemiological data (e.g. range of hosts and geographic distribution) of E. aerophilus in Europe are poorly known, thus at the moment it is difficult to assess to what degree this parasite may be spreading or what influence global warming or other factors may have on the current distribution [31]. On the other hand, it is known that wild foxes act as reservoirs and amplifiers of several canid nematodes, thus they can re-enforce environmental contamination and risk of infection for domestic dogs and humans as well [29].

There is still not an agreement of the taxonomy of capillariid nematodes. Key morphological characters (i.e. eggs, oesophageal structure and genital organs of adult stages) allow the identification of E. aerophilus but, sometimes its identification at the species level and the differentiation from E. boehmi may be difficult.

While Christenson [5] did not detect spicule and spicule sheath in male E. aerophilus, more recent line drawings of this very characteristic organ were published by Butterworth and Beverley-Burton [3], Moravec [17] and Romashov [23]. The present work confirms the presence of these morphological features, which is of key diagnostic relevance for the identification at the species level of male E. aerophilus.

The stichocyte number, higher in E. aerophilus than in E. boehmi, can help to distinguish these two species between them, even though in some cases, the number of stichocytes may overlap. In fact in the present study, one E. boehmi female showed 35 stichocytes as an E. aerophilus female. According to Moravec [17], E. boehmi and E. aerophilus females have 30–32 and 42–46 stichocytes, respectively, whereas, according to Romashov [23], the number of stichocytes in the female is 32–36 for E. boehmi and 35–49 for E. aerophilus. Greater variations of the stichocyte number were reported in E. aerophilus males, i.e. 42–55 [23], or 43–50 [17]. The present study revealed that the number of stichocytes may be useful in the identification of Eucoleus spp. but it is not absolute criteria. The localization of adult stages could also greatly help in distinguishing between E. aerophilus (in trachea and bronchi) and E. boehmi (in nasal cavities).

Another issue warranting further investigations is the size of the eggs. While some textbooks report a length of up to 83 μm for E. aerophilus eggs, e.g. Soulsby [26], Bowman et al. [2], Taylor et al. [28], recent studies have demonstrated that the length of E. aerophilus eggs from dogs and cats is unwaveringly less than 70 μm [7, 8, 32]. Interestingly, some ova collected from the uterus of adult E. aerophilus in the present study were found to be longer and this could be from different reasons. It could be that eggs undergo further development and modifications when they are passed with the faeces, or the existence of different morphotypes within the same species E. aerophilus and with different host affiliations could be hypothesized. Recent molecular studies have shown that different haplotypes of E. aerophilus may be either shared between domestic and wild carnivores or are affiliated to specific hosts [9]. Further studies are necessary for evaluating the distribution of E. aerophilus in wildlife and pets cohabiting the same geographic areas in order to elucidate the phylogeography of different parasite populations and for verifying the possible existence of different morphotypes [8, 9].

Under a clinical standpoint, the pathogenic role of E. aerophilus in wildlife is not well recognized because it is ranging from low to high and E. aerophilus has been also considered as an agent of massive mortality in farmed silver foxes [25]. In the present work, a very low inflammatory response was found around the worms (data not shown) which can be the result of a low pathogenicity or an immunosuppression in the mucosa caused by the parasite in these animals. A past work carried out in six experimentally infected foxes showed that animals may develop bronchopneumonia with cough and other signs of heavy respiratory infection, till death in some cases (Borovkova, cited in [25]). In pets, this parasite may cause bronchovesicular sounds, respiratory inflammation, sneezing, wheezing, chronic moist or dry cough, (broncho)-pneumonia respiratory failure and heavy parasite burdens may lead to mortality [10, 31]. More studies are also necessary to evaluate the actual role of E. aerophilus in causing undiagnosed respiratory distresses in companion dogs and cats.

Another topic which needs to be better elucidated is the actual role of E. aerophilus in causing lung diseases in humans. Indeed, only 12 cases of infection have been published in the literature [13], but it is likely that the infection is underdiagnosed because the clinical signs may overlap a plethora of respiratory diseases which may be self-limiting or may resolve after non-specific treatments.

Acknowledgments

Authors are thankful to Dr. Edoardo Pozio, Istituto Superiore di Sanità, Rome, for critical reading of the manuscript. This study was supported by Ministry of Education and Science of Serbia, Grant No. TR31084.

References

  1. Borgsteede FHM. 1984. Helminth parasites of wild foxes (Vulpes vulpes L.) in The Netherlands. Zeitschrift für Parasitenkunde, 70, 281–285. [CrossRef] [Google Scholar]
  2. Bowman DD, Lynn RC, Eberhard ML. 2003. Georgi’s Parasitology for Veterinarians, 8th edn. St. Louis: Sauders. [Google Scholar]
  3. Butterworth EW, Beverley-Burton M. 1980. The taxonomy of Capillaria spp. (Nematoda: Trichuroidea) in carnivorous mammals from Ontario, Canada. Systematic Parasitology, 1, 211–236. [Google Scholar]
  4. Chautan M, Pontier D, Artois M. 2000. Role of rabies in recent demographic changes in red fox populations in Europe. Mammalia, 64, 391–410. [CrossRef] [Google Scholar]
  5. Christenson RO. 1935. Studies on the morphology of the common fox lungworm, Capillaria aerophila (Creplin, 1839). Transactions of the American Microscopical Society, 54, 145–154. [CrossRef] [Google Scholar]
  6. Davidson RK, Gjerde B, Vikoren T, Lillehaug A, Handeland K. 2006. Prevalence of Trichinella larvae and extra-intestinal nematodes in Norwegian red foxes (Vulpes vulpes). Veterinary Parasitology, 136, 307–316. [CrossRef] [PubMed] [Google Scholar]
  7. di Cesare A, Castagna G, Meloni S, Otranto D, Traversa D. 2012. Mixed trichuroid infestation in a dog from Italy. Parasites & Vectors, 5, 128. [CrossRef] [PubMed] [Google Scholar]
  8. di Cesare A, Castagna G, Otranto D, Meloni S, Milillo P, Latrofa MS, Paoletti B, Bartolini R, Traversa D. 2012. Molecular detection of Capillaria aerophila, an agent of canine and feline pulmonary capillariosis. Journal of Clinical Microbiology, 50, 1958–1963. [CrossRef] [PubMed] [Google Scholar]
  9. di Cesare A, Otranto D, Latrofa MS, Meloni S, Castagna G, Morgan E, Lalosevic D, Mihalca A, Padre L, Traversa D. 2012. Genetic characterization of Eucoleus aerophilus from different hosts and countries. in: Proceedings of the 27th Conference of the Italian Society of Parasitology (SO.I.PA.), 26th–29th June, Alghero, Italy. [Google Scholar]
  10. Foster S, Martin P. 2011. Lower respiratory tract infections in cats. Journal of Feline Medicine and Surgery, 13, 313–332. [CrossRef] [PubMed] [Google Scholar]
  11. König A, Romig T, Thoma D, Kellermann K. 2005. Drastic increase in the prevalence of Echinococcus multilocularis in foxes (Vulpes vulpes) in southern Bavaria, Germany. European Journal of Wildlife Research, 51, 277–282. [CrossRef] [Google Scholar]
  12. Laloševic D, Dimitrijevic S, Jovanovic M, Klun I. 2001. Pulmonary aelurostrongylosis in cats. Veterinarski Glasnik, 55, 181–185 (in Serbian). [Google Scholar]
  13. Laloševic D, Laloševic V, Klem I, Stanojev-Jovanovic D, Pozio E. 2008. Pulmonary capillariasis miming bronchial carcinoma. American Journal of Tropical Medicine and Hygiene, 78, 14–16. [Google Scholar]
  14. Magi M, Macchioni F, dell’Omodarme M, Prati MC, Calderini P, Gabrielli S, Iori A, Cancrini G. 2009. Endoparasites of red fox (Vulpes vulpes) in central Italy. Journal of Wildlife Disease, 45, 881–885. [Google Scholar]
  15. Miterpáková M, Hurnĺková Z, Antolová D, Dubinský P. 2009. Endoparasites of red fox (Vulpes vulpes) in the Slovak Republic with the emphasis on zoonotic species Echinococcus multilocularis and Trichinella spp. Helminthologia, 46, 73–79. [CrossRef] [Google Scholar]
  16. Moravec F. 1982. Proposal of a new systematic arrangement of nematodes of the family Capillariidae. Folia Parasitologica (Praha), 29, 119–132. [Google Scholar]
  17. Moravec F. 2000. Review of capillariid and trichosomoidid nematodes from mammals in the Czech Republic and the Slovak Republic. Acta Societatis Zoologicae Bohemicae, 64, 271–304. [Google Scholar]
  18. Moravec F, Prokopic J, Shlikas AV. 1987. The biology of nematodes of the family Capillariidae Neveu-Lemaire, 1936. Folia Parasitologica (Praha), 34, 39–56. [Google Scholar]
  19. Nithikathkul C, Saichua P, Royal L, Cross JH. 2011. Capillariosis, in Oxford Textbook of Zoonoses, 2nd Edition, Biology, Clinical Practice, and Public Health Control, Palmer SR, Lord Soulsby EJL, Torgerson P, Brown DWG, Editors. Oxford University Press: Oxford. p. 727–737. [Google Scholar]
  20. Rajkovic-Janje R, Marinculic A, Bosnic S, Benic M, Vinkovic B, Mihaljevic Z. 2002. Prevalence and seasonal distribution of helminth parasites in red foxes (Vulpes vulpes) from the Zagreb County (Croatia). Zeitschrift für Jagdwissenschaft, 48, 151–160. [Google Scholar]
  21. Reperant LA, Hegglin D, Fischer C, Kohler L, Weber JM, Deplazes P. 2007. Influence of urbanization on the epidemiology of intestinal helminths of the red fox (Vulpes vulpes) in Geneva, Switzerland. Parasitology Research, 101, 605–611. [CrossRef] [PubMed] [Google Scholar]
  22. Robardet E, Giraudoux P, Caillot C, Boue F, Cliquet F, Augot D, Barrat J. 2008. Infection of foxes by Echinococcocus multilocularis in urban and suburban areas of Nancy, France: influence of feeding habits and environment. Parasite, 15, 77–85. [CrossRef] [EDP Sciences] [PubMed] [Google Scholar]
  23. Romashov BV. 2000. Three capillariid species (Nematoda, Capillariidae) of carnivores (Carnivora) and discussion of system and evolution of the nematode family Capillariidae. 1. Redescription of Eucoleus aerophilus and E. boehmi. Zoologichesky Zhurnal, 79, 1379–1391 (in Russian). [Google Scholar]
  24. Simpson VR. 2002. Wild animals as reservoirs of infectious diseases in the UK. The Veterinary Journal, 163, 128–146. [CrossRef] [Google Scholar]
  25. Skryabin KI, Shihobalova NP, Orlov IV. 1957. Trichocephalids and capillariids of animals and humans, and the disease caused by them, Fundamentals of nematodology, Vol VI. Akademiya Nauk SSSR: Moscow. p. 536–548 (in Russian). [Google Scholar]
  26. Soulsby EJL. 1982. Helminths, arthropods and protozoa of domesticated animals, 7th edition. Philadelphia: Lea & Febiger. [Google Scholar]
  27. Sréter T, Széll Z, Marucci G, Pozio E, Varga I. 2003. Extraintestinal nematode infections of red foxes (Vulpes vulpes) in Hungary. Veterinary Parasitology, 115, 329–334. [CrossRef] [PubMed] [Google Scholar]
  28. Taylor MA, Coop RL, Wall RL. 2007. Veterinary parasitology, 3rd edn. Oxford: Blackwell Publishing Ltd. [Google Scholar]
  29. Traversa D. 2012. Pet roundworms and hookworms:Acontinuing need for global warming. Parasites & Vectors, 5, 91. [CrossRef] [PubMed] [Google Scholar]
  30. Traversa D, di Cesare A, Milillo P, Iorio R, Otranto D. 2009. Infection by Eucoleus aerophilus in dogs and cats: is another extra-intestinal parasitic nematode of pets emerging in Italy? Research in Veterinary Science, 87, 270–272. [CrossRef] [PubMed] [Google Scholar]
  31. Traversa D, di Cesare A, Conboy G. 2010. Canine and feline cardiopulmonary parasitic nematodes in Europe: emerging and underestimated. Parasites & Vectors, 3, 62. [CrossRef] [PubMed] [Google Scholar]
  32. Traversa D, di Cesare A, Lia RP, Castagna G, Meloni S, Heine J, Strube K, Milillo P, Otranto D, Meckes O, Schaper R. 2011. New insights into morphological and biological features of Capillaria aerophila (Trichocephalida, Trichuridae). Parasitology Research, 109 (suppl. 1), S97–S104. [CrossRef] [PubMed] [Google Scholar]
  33. Vos AC. 1995. Population dynamics of the red fox (Vulpes vulpes) after the disappearance of rabies in county Garmisch- Partenkirchen, Germany, 1987–1992. Annales Zoologici Fennici, 32, 93–97. [Google Scholar]

Cite this article as: Lalošević V, Laloševicć D, Čapo I, Simin V, Galfi A & Traversa D: High infection rate of zoonotic Eucoleus aerophilus infection in foxes from Serbia. Parasite, 2013, 20, 3.

All Tables

Table 1.

Results for the copromicroscopic examinations of the foxes (n = 118) examined in the present study. N: number of foxes scored positive for endoparasites.

All Figures

thumbnail Figure 1.

Adult female Eucoleus aerophilus on the mucosa of the opened trachea of a fox dissected after formalin fixation. More than half of the worm body is inside the tracheal mucosa with a zig-zag shape and the other part is free in the lumen. Bar, 1 mm.

In the text
thumbnail Figure 2.

Number of Eucoleus aerophilus adults per foxes.

In the text
thumbnail Figure 3.

Female Eucoleus aerophilus showing the end part of the last stichocyte with the cell nucleus (N), two superimposed secretory cells (S), intestine (I), vagina with eggs (E) and vulva (arrowhead). Bar, 50 μm.

In the text
thumbnail Figure 4.

Male Eucoleus aerophilus, extruded spinose spicule sheath and partially extruded spicule at the caudal part. Bar, 100 μm.

In the text

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