Next Article in Journal
Prolactin Expression in the Baboon (Papio hamadryas) Eye
Next Article in Special Issue
Microsporidia as a Potential Threat to the Iberian Lynx (Lynx pardinus)
Previous Article in Journal
Propagation of Babesia bigemina in Rabbit Model and Evaluation of Its Attenuation in Cross-Bred Calves
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Animals
Volume 12
Issue 17
10.3390/ani12172289
animals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

A High Prevalence of Cardiopulmonary Worms Detected in the Iberian Wolf (Canis lupus): A Threat for Wild and Domestic Canids

by
Efrén Estévez-Sánchez
1,
Rocío Checa
1,
Ana Montoya
1,
Juan Pedro Barrera
1,
Ana María López-Beceiro
2,
Luis Eusebio Fidalgo
2 and
Guadalupe Miró
1,*
1
Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Avda. Puerta de Hierro s/n, 28040 Madrid, Spain
2
Departamento de Anatomía, Producción Animal y Ciencias Clínicas Veterinarias, Facultad de Veterinaria, Universidad de Santiago de Compostela, Campus Terra, 27002 Lugo, Spain
*
Author to whom correspondence should be addressed.
Animals 2022, 12(17), 2289; https://doi.org/10.3390/ani12172289
Submission received: 27 July 2022 / Revised: 27 August 2022 / Accepted: 31 August 2022 / Published: 3 September 2022
(This article belongs to the Special Issue The Epidemiology, Diagnosis and Prevention of Infectious Diseases in Wildlife)
Browse Figures
Review Reports Versions Notes

Abstract

:

Simple Summary

The Iberian wolf (Canis lupus signatus) is a recognized reservoir of some zoonotic parasites that cause diseases transmissible to domestic animals and/or humans. The objective of this study was to determine the diversity of species of cardiopulmonary nematode parasites that affect wolves in northwestern Spain, and to estimate their prevalence and the relationship between these parasites and various epidemiological variables. The cardiopulmonary systems of 57 wolves from Galicia were examined using dissection and cup sedimentation techniques, and the collected worms were then morphologically identified. The overall prevalence of infection by cardiopulmonary nematodes was 24.5%, and the parasite species identified were Angiostrongylus vasorum (19.3%), Crenosoma vulpis (7%) and Eucoleus aerophilus (3.5%). The latter is of zoonotic interest. A significant relationship was found between age and C. vulpis infection, which was only found in animals under one year of age. Our findings indicate that the Iberian wolf could play an important role in maintaining cardiopulmonary parasites in the wild, and they highlight a need to identify potential risks for veterinary and public health.

Abstract

Cardiopulmonary nematodes are highly pathogenic parasites affecting domestic and wild canids. As the result of conservation programs, the Iberian wolf (Canis lupus signatus) population has recently expanded, and its distribution range covers lands from where it had long disappeared. However, the exact epidemiological role of the wolf in the life cycle of zoonotic parasites causing diseases transmissible to pets and/or humans is largely unknown. This study sought to determine the diversity of cardiopulmonary nematode parasite species that affect wolves inhabiting northwestern areas of the Iberian Peninsula, and to estimate their prevalence and the relationship between these parasites and several epidemiological variables. For this purpose, we examined the cardiopulmonary systems of 57 wolves from Galicia (from the provinces A Coruña n = 15, Lugo n = 21, Ourense n =15 and Pontevedra n = 6) using techniques of dissection and cup sedimentation. Collected worms were then identified under a light microscope according to their morphological features. Three species of nematodes were detected: Angiostrongylus vasorum (the “French-heartworm”), Crenosoma vulpis and Eucoleus aerophilus, the latter being of zoonotic interest. The prevalence was 24.5% (14/57; 95% CI 13.3–35.6%) overall, 19.3% for A. vasorum (11/57; 95% CI 8.8–29.2%), 7% for C. vulpis (4/57; 95% CI 0.4–13.6%) and 3.5% for E. aerophilus (2/57; CI −1.1–9.1%). A significant relationship (p = 0.002) was found between age and the presence of C. vulpis, which was only found in juvenile animals. Furthermore, a higher prevalence of A. vasorum and/or C. vulpis was observed in wolves with a lower body condition score (40% and 20%, respectively), though the difference was not significant (p = 0.221 and p = 0.444, respectively). Our findings indicate a high “French-heartworm” and lungworm burden in the wolf population of northern Spain, and they identify a need for studies designed to elucidate the epidemiological role played by the Iberian wolf and to identify possible risks for veterinary and public health.

Graphical Abstract

1. Introduction

The Iberian wolf (Canis lupus signatus) is the second largest predator in the Iberian Peninsula, only surpassed by the Eurasian brown bear (Ursus arctos arctos). With an estimated population of 1700 to 2500 wolves in Spain [1], many inhabit its northwestern corner, comprising an area of 120,000 km2, mainly spanning the autonomous communities of Galicia, Asturias and Castilla-Leon [2]. The habitats of these carnivores are often anthropic rural areas [3] where they can feed on both wild and domestic ungulates. This added to the recent expansion of the wolf population across the Iberian Peninsula has rekindled the old debate about what should be done about these feared hunters.
Wolves play an important ecological and health role in the natural environment. They are predators of sick animals, preventing the transmission of diseases such as tuberculosis [4], or promoting vegetation regrowth by reducing the ungulate population [5]. However, they are also an important component of the life cycle of many emerging parasites, some of which are highly pathogenic and potentially lethal for companion animals and humans.
Parasitic diseases take on a special interest because of their possible impact on public health, but also for the conservation and protection of their host species [6]. After many years of recovery, wolf populations have expanded to inhabit areas throughout the Iberian Peninsula [7]. However, because of their wildlife style, it is difficult to preserve their health state and prevent diseases that threaten their conservation. Cardiopulmonary nematode worms cause chronic, usually subclinical diseases in domestic animals, but wolves require their full lung and heart capacity for daily life [8]. Therefore, these diseases can be fatal, especially in young animals.
Owing to their emergent nature and spread to thus far non-endemic places, some cardiopulmonary parasites, such as the metastrongyloids Angiostrongylus vasorum and Crenosoma vulpis, the tricurid Eucoleus aerophilus (syn. Capillaria aerophila), the spirurid Dirofilaria immitis and the family Filaroididae (Oslerus osleri and Filaroides hirthi), are starting to attract the interest of scientists. The exact cause of the increase in parasite numbers is unknown, but global warming, changes in vector seasonal population dynamics and movements in animal populations are thought to play an important role in this expansion [9].
Angiostrongylus vasorum (“French-heartworm”) and Crenosoma vulpis (lungworm) have an indirect biological cycle and affect wild carnivores such as the pine marten (Martes martes), red fox (Vulpes vulpes) and wolf (Canis lupus) [10,11,12]. The intermediate host is a land or aquatic gastropod [13] that spreads third-stage larvae (L3) by ingestion. The life of these gastropods is dependent on climate conditions of humidity and temperature, and they hibernate or aestivate (depending on the species) when these conditions become too harsh [14].
The lungworm Eucoleus aerophilus is a parasite affecting dogs, cats, wild carnivores and, sometimes, humans worldwide [15]. Its life cycle is not known exactly, but it seems to be transmitted directly by the fecal–oral route or through paratenic hosts (earthworms) harboring L1 larvae [16].
Dirofilaria immitis is a zoonotic parasite that mainly affects dogs and cats, although it can infest several mammals. Dirofilariosis is transmitted by mosquitoes (Diptera, Culicidae) and is endemic in southern Mediterranean countries including Spain [17].
The lungworms Oslerus osleri and Filaroides hirthi both affect canids. Those of the family Filaroididae are normally parasites of immunocompromised, stressed or young dogs, and only few cases have been reported in domestic animals in Spain [18,19,20]. Transmission from mothers to their puppies occurs via licking or through coprophagy [21].
Despite being such a relevant topic, few studies have addressed the epidemiology and pathogenesis of cardiopulmonary parasites in wolves in Europe. Spain is currently considered an endemic area, and the red fox (Vulpes vulpes) seems to be the main reservoir of these parasites [8,10,12]. Although their prevalence in Spanish domestic dogs is apparently unrepresentative (0.75–1.73%) [22,23], the life cycle of cardiopulmonary parasites in wild hosts has been established, affecting animals such as wolves, foxes and other mesocarnivores. As a result of this close relationship between the domestic and wild life cycle, these reservoirs could play a key role in parasite spread to pets and humans [24].
The aim of this study was to examine the presence of cardiopulmonary parasites in wolf populations of the northwestern Iberian Peninsula and to analyze the different epidemiological variables involved.

2. Materials and Methods

2.1. Study Area

The study population comprised 57 Iberian wolves (Canis lupus signatus) from northwest Spain. These wolves died of natural causes or were killed in road accidents in the Galicia region, and they were collected by official organisms of Galicia over the period of 2016 to 2021. In this geographical area, climate is predominantly oceanic, with annual temperature averages of around 12.0–14.9 °C and rainfall averages of around 811–1791 mm in 2021 [25].

2.2. Sample and Data Collection

The following data were collected for each animal: age, sex, weight, body condition score and any other relevant findings in a physical examination. Age was classified according to body weight, dentition and sexual maturity, as: wolf pups (under 12 months of age and without full body development), immature or young (1 to 3 years of age and not yet sexually mature) and adults (over 3 years, or of reproductive age). Body condition was scored from 1 to 5, where 1 indicates extreme thinness, 3 indicates an optimal state and 5 is markedly overweight.
Wolves were necropsied following standard procedures. Cardiorespiratory tissue samples were obtained by sectioning the neck muscles and dissecting adjacent tissues to harvest the laryngeal cartilages, trachea, lungs and heart in a single block. Biological samples were stored frozen (−20 °C) until their laboratory analysis.

2.3. Cardiorespiratory Parasites

The trachea, bronchi and bronchioles were longitudinally dissected and inspected to collect as many nematodes as possible. The heart chambers and blood vessels were also carefully dissected following the blood circulation. Once the lungs and heart were fully dissected, they were deposited in a tapered graduated sedimentation cup. The cups were filled with warm water until the organs were completely submerged and set to rest for at least 6 h to promote worm sedimentation. The sediment was filtered, and the remaining material was examined under a stereomicroscope to identify the worms [26,27].
All worms collected during this procedure were deposited in tubes containing 70% ethanol for less than 72 h. Subsequently, the parasites were deposited in a glass dish and identified morphologically under a light microscope. Worms were classified at the species level according to sex (male or female) using identification keys [16,28,29,30]. The total number of parasites collected from each cardiorespiratory system was also recorded.

2.4. Statistical Analysis

The prevalence (percentage of infected animals), parasite intensity (number of parasites per infected animal) and species richness (number of different species parasitizing the same host) were reported. The differences observed according to epidemiological variables (age, sex, body condition and province) and the significance of each association (between prevalence and each epidemiological variable) were analyzed using Fisher’s exact test at a 95% confidence interval (significance was set at p ≤ 0.05). The software employed for this analysis was R version 4.2.1.

3. Results

3.1. Nematode Parasite Species and Prevalence

The overall proportion of wolves detected positive was 24.5% (14/57; 95% CI 13.3–35.6%). The species identified were Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus. Other species such as Dirofilaria immitis, Oslerus osleri and Filaroides hirthi were not found by necropsy in these animals.
The individual prevalence was: A. vasorum 19.3% (11/57; 95% CI 8.8–29.2%), C. vulpis 7% (4/57; 95% CI 0.4–13.6%) and E. aerophilus 3.5% (2/57; CI −1.1%–9.1%). Eleven (19.3%) of the wolves were infected by one species, and three (5.2%) had a two-species coinfection (Table 1). No animal was infected by the three different species.
The total number of worms collected was 385, with 275 females (71.4%) and 72 males (18.7%). Sex could not be determined in 38 worms. The number of parasites found in the post mortem exam of a single infected wolf ranged from 1 to 99 parasites (mean 27.5 ± 33.4). Furthermore, in the case of A. vasorum, the number of worms per animal (single infections and co-infections) ranged from 1 to 75 and from 1 to 24 for female and male worms, respectively.
All wolves infected by A. vasorum had at least one female worm (11/11), and 82% had at least one male worm (9/11). In addition, 25% of the animals infected by C. vulpis had one male worm (1/4), and only female worms were identified in animals harboring E. aerophilus (Table 1).

3.2. Epidemiological Data Recorded in Wolves with Nematode Worms

Of the 57 Iberian wolves examined, 21 were from the Galician province of Lugo (37%), 15 were from A Coruña (26%), 15 were from Ourense (26%) and 6 were from Pontevedra (11%). A total of 29 were male (51%), and 28 were females (49%). A total of 14 were classified as wolf pups (25%), 24 were classified as young (24%) and 19 were classified as adults and older adult wolves (33%). For the whole population, body condition scores (out of 5) were: 22% 1–2 (n = 12), 35% 3 (n = 20) and 45% 4–5 (n = 24).
A higher overall worm prevalence was observed in the A Coruña and Ourense provinces (26.7%), followed by Lugo (23.8%) and Pontevedra (16.7%) (Figure 1). By species, the higher prevalence of A. vasorum was detected in Lugo (23.8%), whereas infection by C. vulpes was only observed in A Coruña and Ourense (13.3%) and by E. aerophilus in the provinces of Ourense and Lugo (6.7% and 4.8%, respectively) (Table 2). Notwithstanding, differences in cardiopulmonary worm prevalence between provinces were not significant (p = 1).
Nearly similar overall prevalence was detected in males and females, with 24.1% and 25%, respectively. However, infection by A. vasorum, C. vulpis and E. aerophilus was higher in females (21.4%, 7.1% and 7.1%, respectively), although these differences were not significant (Table 2).
Wolves with a lower body condition score showed a higher prevalence of A. vasorum and/or C. vulpis (40% and 20%, respectively), yet the difference was once again not significant. However, animal age and the presence of C. vulpis were significantly associated, such that this lungworm species was only found in juvenile animals (p = 0.002).
To compare worm infection intensity according to host age, nematode numbers were grouped into four categories. p-values were calculated comparing an age group of wolves with the remaining wolves for each grade of parasitization (Table 3). Hence, all infected wolf pups (42.9%) had fewer than 30 parasites per animal, and among the infected young wolves and adults, 8.3% and 5.26% had fewer than 30 parasites, 8.3% and 5.26% had between 30 and 60 parasites and 4.2% and 5.26% had more than 60 parasites, respectively. A significant link was thus observed between a low number of parasites (less than 30) and a less than one-year wolf age (p = 0.004).

4. Discussion

All three cardiopulmonary nematodes identified in our study (A. vasorum, C. vulpis and E. aerophilus) have been previously described in wolves from the Iberian Peninsula [6,8,11,12,31] and from other European countries [16,32,33,34,35,36]. It should also be noted that O. osleri, F. hirthi and D. immitis, reported previously in wolves from northwestern Spain, were not found in the present study. For Europe, few studies have provided lungworm and heartworm prevalence data in wolves. A. vasorum has been described in wolves from Slovakia (0.8%) [32], Croatia (3.1%) [33] and Italy (28%). Compared to Spain, C. vulpis is more prevalent in wolves from countries such as Latvia (9.1%) [35], Portugal (9.1%) [6] and Belarus (7.7%). Similarly, E. aerophilus shows a high prevalence in Europe (9–36%) [6,32,33,35].
There are mainly four techniques used for the detection of cardiopulmonary parasites: Baermann migration–sedimentation, immunological diagnosis through antigen- or antibody-based assays, molecular detection (polymerase chain reaction) and dissection identification [37,38,39,40,41]. This determines that the reported prevalence data are difficult to interpret because of the different sensitivity and specificity of the diagnostic techniques used. For wild animals, feces samples collected directly from the ground are often used. However, first-stage A. vasorum and C. vulpis larvae (L1) are susceptible to environmental conditions, and these studies are likely underestimating the prevalence. In contrast, Eucoleus aerophilus releases very resistant eggs [9], so data for this nematode could be considered more representative. This makes it difficult to make any comparison of prevalence data between different countries or regions. Furthermore, this study has some potential limitations: first, not all wolves were suitable for necropsy because of the bad preservation state of the tissues, and the limited sample size could have affected the statistical analysis; second, no other diagnostic tests have been performed to compare the results due to the lack of samples.
Our prevalence data for A. vasorum (19.3%) were similar to the reported rates based also on anatomical dissection in wolves from northwestern Spain (2.1–22%) [8,12]. These rates are lower than the A. vasorum prevalence reported in red foxes (Vulpes vulpes) (16–43.2%) [8,42,43,44,45], similar to those described for the Eurasian badger (Meles meles) (6.4–24%) [45,46], and higher than those provided for dogs (0.73–1.73%) [22,23] in Spain based on serology. Dogs do not usually feed on intermediate hosts [22], and wolves prefer larger prey [13]. However, terrestrial gastropods (intermediate hosts), anurans and birds (paratenic hosts) are a common food source for the fox and badger, and this is an accepted explanation for the different prevalence found among species [45].
In foxes, A. vasorum infection does not produce an efficient immune response, so adults previously exposed to this parasite can become reinfected and even sustain a persistent infection [47]. Although no study has been carried out in wolves, the similar prevalence detected here among juveniles and adults (21.7% and 23.3%, respectively) could mean that wolves also have no efficient immune response against this parasite. To better understand this, we should consider that wolves younger than one year showed an overall higher prevalence of cardiopulmonary parasites (42.9%) compared to young wolves (between 1 and 3 years) (28.8%), and in the specific case of A. vasorum, a similar prevalence was detected at both ages (21.4% and 20.8%, respectively). This means that only in the case of A. vasorum is there a stable (slightly upward) prevalence trend with animal age. In addition, young and adult animals showed a greater extent of parasitization with A. vasorum, in that 13.56% showed moderate infection, and 9.46% showed severe infection, which means that they could have persistent infections and/or be continuously reinfected.
Angiostrongylosis is an emerging disease causing severe respiratory and central nervous system signs in dogs that has spread throughout central and northwestern Europe. It is today considered endemic in many countries. Recent reports demonstrate that canine A. vasorum is expanding and that new foci are appearing in previously non-endemic areas [48]. Although the red fox is thought to be responsible for this expansion, the wolf may also play an important epidemiological role as a reservoir in the wildlife cycle, infecting other wild or domestic animals. In the last two decades, an increase in the prevalence of A. vasorum infection has been reported in the wolf population of the northwestern Iberian Peninsula from 2% [12] to 22–24.5% [8]. Spain is currently considered a stable focus of a high prevalence of A. vasorum due to its conditions of temperature and moisture, which facilitate the survival of intermediate hosts and first-stage larvae (L1) in the environment [49].
In Spain, Crenosoma vulpis has been described in the wolf, red fox and dog. The prevalence in wolves detected here (7%) was slightly lower than in previous studies carried out in the northwestern Iberian Peninsula (9.1–9.4%) [6,8]. The red fox is the main reservoir of C. vulpis in the Iberian Peninsula, and its prevalence is significantly higher than in the wolf (2.5–44.8%) [8,42,44,49]. To our knowledge, only one study has provided prevalence data for C. vulpis in Spanish dogs (2%) [50]. Our results reveal differences between C. vulpis and A. vasorum in wolves: (i) C. vulpis was only detected in juvenile wolves (28.6%), with the difference with older animals being significant (p = 0.002). This could be explained by the smaller prey caught by young wolves, but it could also reflect an efficient immune response against C. vulpis in adult wolves; and (ii) C. vulpis has been described in geographical areas lacking the presence of A. vasorum or where cases are only sporadic. Examples of these regions are Canada, Finland, Norway, Ireland or the United Kingdom [51], as first-stage C. vulpis larvae (L1) can remain active at low temperatures even after a freezing period of several days [52].
When infected wolves were classified according to body condition, those described as mildly thin were found to harbor the highest prevalence of parasites (50%). Thus, 40% of individuals with this body condition were parasitized with A. vasorum, 20% were parasitized with C. vulpis and none were parasitized with E. aerophilus. Thus, it seems that the most affected animals were infected by A. vasorum and C. vulpis, as these parasites, respectively, attack the vascular endothelium [11] and have an irritant effect on the respiratory system [12].
Eucoleus aerophilus has been described in both domestic and wild carnivores from Spain [31,43,46,53]. The prevalence of this nematode in wolves in northwestern Spain (4%) was lower than previously described (5.4–50.54%) [8,54]. In Spain, E. aerophilus is also more prevalent in the red fox (4.4–34.2%) [8,42,55,56] and pine marten (Martes martes) (50.98%) [10], but studies assessing this infection in domestic animals have been scarce. Unlike A. vasorum and C. vulpis, E. aerophilus plays a direct role in the life cycle for which the ingestion of eggs or paratenic hosts such as lumbricids are required [24]. However, its exact biological life cycle is unknown, so more studies are needed to understand the impact of these results.
The role of terrestrial gastropods and earthworms is of paramount importance in the distribution and prevalence of cardiopulmonary parasites. This relationship can be observed when comparing earthworm and mollusc distribution maps [57,58] with endemic areas of the parasites throughout Europe. It may be observed that Pontevedra has a lower abundance of earthworms and terrestrial gastropods, possibly explaining the lower prevalence of cardiopulmonary parasites detected in this province compared to the others included in the present study.
In conclusion, our observations indicate that the Iberian wolf could play an important role in the maintenance of cardiopulmonary parasites in the wild. Some studies have confirmed the lack of segregation of parasites among foxes, coyotes and dogs [59], so transmission occurs because of proximity between the wild and domestic life cycle. In addition, wolves are susceptible to the same infectious and parasitic diseases [60] as the other domestic and wild canids. We would like to highlight the need to clarify the role played by the Iberian wolf in these parasitoses and to identify the risks they could pose for animal and human health.

Author Contributions

Conceptualization, R.C., A.M., A.M.L.-B. and G.M.; Data curation, E.E.-S. and A.M.; Formal analysis, R.C. and A.M.L.-B.; Investigation, R.C., J.P.B., A.M.L.-B., L.E.F. and G.M.; Methodology, E.E.-S., J.P.B., A.M.L.-B. and L.E.F.; Writing—original draft preparation, E.E.-S. and R.C.; Writing—review and editing, A.M., A.M.L.-B., L.E.F. and G.M. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

This study was carried out in accordance with the Spanish legislation guidelines (RD 8/2003) and with the international Guiding Principles for Biomedical Research Involving Animals issued by the Council for International Organization of Medical Sciences and the International Council for Laboratory Animal Science (RD 53/2013). Ethical review and approval were waived as the samples for this study were obtained from animals that had died of natural causes.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Acknowledgments

Wild Fauna Recovery Centres of Galicia; Dirección Xeral de Patrimonio Natural (Xunta de Galicia, Spain).

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Arija, C.M. Biología y Conservación del Lobo Ibérico: Crónica de un conflicto. REDVET. Rev. Electrónica de Vet. 2010, 11, 1–18. [Google Scholar]
  2. Blanco, J.C.; Saénz de Buruaga, M.; Llaneza, L. Canis Lupus Linnaeus, 1758. In Atlas y Libro Rojo de mamíferos terrestres de España; Palomo, L.J., Gisbert, J., Blanco, J.C., Eds.; Dirección General para la Biodiversidad -SECEM-SECEMU: Madrid, Spain, 2007; pp. 272–276. [Google Scholar]
  3. Blanco, J.C. La Gestión Del Lobo En España. Controversias Científicas En Torno a Su Caza. arbor 2017, 193, a418. [Google Scholar] [CrossRef]
  4. Tanner, E.; White, A.; Acevedo, P.; Balseiro, A.; Marcos, J.; Gortázar, C. Wolves Contribute to Disease Control in a Multi-Host System. Sci. Rep. 2019, 9, 7940. [Google Scholar] [CrossRef] [PubMed]
  5. Blanco, J.C.; Cortés, Y. Ecología, Censos, Percepción y Evolución Del Lobo En España: Análisis de Un Conflicto; SECEM: Málaga, Spain, 2002; ISBN 84-607-3896-5. [Google Scholar]
  6. Figueiredo, A.; Oliveira, L.; de Carvalho, L.M.; Fonseca, C.; Torres, R.T. Parasite Species of the Endangered Iberian Wolf (Canis lupus signatus) and a Sympatric Widespread Carnivore. Int. J. Parasitol. Parasites Wildl. 2016, 5, 164–167. [Google Scholar] [CrossRef]
  7. Viejo, J.L.; Montesinos, V. La Fauna de La Sierra de Guadarrama. Somera Aproximación a La Riqueza Faunística de Un Nuevo Parque Nacional. Ambienta 2019, 103, 2–25. [Google Scholar]
  8. Martínez-Rondán, F.J.; de Ybáñez, M.R.R.; López-Beceiro, A.M.; Fidalgo, L.E.; Berriatua, E.; Lahat, L.; Sacristán, I.; Oleaga, Á.; Martínez-Carrasco, C. Cardiopulmonary Nematode Infections in Wild Canids: Does the Key Lie on Host-Prey-Parasite Evolution? Res. Vet. Sci. 2019, 126, 51–58. [Google Scholar] [CrossRef]
  9. Traversa, D.; Di Cesare, A.; Conboy, G. Canine and Feline Cardiopulmonary Parasitic Nematodes in Europe: Emerging and Underestimated. Parasites Vectors 2010, 3, 62. [Google Scholar] [CrossRef]
  10. 10 Segovia, J.-M.; Torres, J.; Miquel, J.; Sospedra, E.; Guerrero, R.; Feliu, C. Analysis of Helminth Communities of the Pine Marten, Martes martes, in Spain: Mainland and Insular Data. Acta Parasitol. 2007, 52, 156–164. [Google Scholar] [CrossRef]
  11. Balmori, A.; Rico, M.; Naves, J.; Llamazares, E. Contribución al Estudio de Los Endoparásitos Del Lobo En La Península Ibérica: Una Investigación Coprológica. Galemys 2000, 12, 13–26. [Google Scholar]
  12. Segovia, J.; Torres, J.; Miquel, J.; Llaneza, L.; Feliu, C. Helminths in the Wolf, Canis lupus, from North-Western Spain. J. Helminthol. 2001, 75, 183–192. [Google Scholar]
  13. Ferdushy, T.; Hasan, M.T. Angiostrongylus vasorum: The ‘French Heartworm. Parasitol. Res. 2010, 107, 765–771. [Google Scholar] [CrossRef] [PubMed]
  14. Tolnai, Z.; Széll, Z.; Sréter, T. Environmental Determinants of the Spatial Distribution of Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus in Hungary. Vet. Parasitol. 2015, 207, 355–358. [Google Scholar] [CrossRef] [PubMed]
  15. Traversa, D.; Di Cesare, A.; Lia, R.; Castagna, G.; Meloni, S.; Heine, J.; Strube, K.; Milillo, P.; Otranto, D.; Meckes, O.; et al. New Insights into Morphological and Biological Features of Capillaria aerophila (Trichocephalida, Trichuridae). Parasitol. Res. 2011, 109 (Suppl. 1), S97–S104. [Google Scholar] [CrossRef]
  16. Bowman, D. Georgis’ Parasitology for Veterinarians, 9th ed.; Elsevier España: St. Louis, MI, USA, 2011; ISBN 84-8086-705-1. [Google Scholar]
  17. Miró, G.; Montoya, A.; Roura, X.; Gálvez, R.; Sainz, A. Seropositivity Rates for Agents of Canine Vector-Borne Diseases in Spain: A Multicentre Study. Parasites Vectors 2013, 6, 117. [Google Scholar] [CrossRef] [Green Version]
  18. Caro-Vadillo, A.; Martínez-Merlo, E.; García-Real, I.; Fermín-Rodríguez, M.L.; Mateo, P. Verminous Pneumonia Due to Filaroides hirthi in a Scottish Terrier in Spain. Vet. Rec. 2005, 157, 586–589. [Google Scholar] [CrossRef] [PubMed]
  19. Luengo, M.; Arata, N. Filaroides osleri as a Finding in Canine Necropsy. Bol. Chil. Parasitol. 1970, 25, 87–88. [Google Scholar]
  20. Carrasco, L.; Herva’s, J. Massive Filaroides hirthi Infestation Associated with Caninedistemper. Vet. Rec. 1997, 140, 72–73. [Google Scholar] [CrossRef]
  21. Georgi, J.R.; Georgi, M.E.; Fahnestock, G.R.; Theodorides, V.J. Transmission and Control of Filaroides hirthi Lungworm Infection in Dogs. Am. J. Vet. Res. 1979, 40, 829–831. [Google Scholar]
  22. Carretón, E.; Morchón, R.; Falcón-Cordón, Y.; Matos, J.; Costa-Rodríguez, N.; Montoya-Alonso, J.A. First Epidemiological Survey of Angiostrongylus vasorum in Domestic Dogs from Spain. Parasites Vectors 2020, 13, 306. [Google Scholar] [CrossRef]
  23. Morchón, R.; Montoya-Alonso, J.A.; Sánchez-Agudo, J.Á.; de Vicente-Bengochea, J.; Murcia-Martínez, X.; Carretón, E. Angiostrongylus vasorum in Domestic Dogs in Castilla y León, Iberian Peninsula, Spain. Animals 2021, 11, 1513. [Google Scholar] [CrossRef]
  24. Mathews, F. Zoonoses in Wildlife: Integrating Ecology into Management. Adv. Parasitol. 2009, 68, 185–209. [Google Scholar] [PubMed]
  25. Agencia Estatal de Meteorología (AEMET) Atlas Climático. Available online: http://agroclimap.aemet.es/# (accessed on 21 July 2020).
  26. Schug, K.; Krämer, F.; Schaper, R.; Hirzmann, J.; Failing, K.; Hermosilla, C.; Taubert, A. Prevalence Survey on Lungworm (Angiostrongylus vasorum, Crenosoma vulpis, Eucoleus aerophilus) Infections of Wild Red Foxes (Vulpes vulpes) in Central Germany. Parasites Vectors 2018, 11, 85. [Google Scholar] [CrossRef]
  27. Schnyder, M.; Fahrion, A.; Ossent, P.; Kohler, L.; Webster, P.; Heine, J.; Deplazes, P. Larvicidal Effect of Imidacloprid/Moxidectin Spot-on Solution in Dogs Experimentally Inoculated with Angiostrongylus vasorum. Vet. Parasitol. 2009, 166, 326–332. [Google Scholar] [CrossRef] [PubMed]
  28. Conboy, G. Helminth Parasites of the Canine and Feline Respiratory Tract. Vet. Clin. N. Am. Small Anim. Pract. 2009, 39, 1109–1126. [Google Scholar] [CrossRef] [PubMed]
  29. Anderson, R.C.; Chabaud, A.G.; Willmott, S. Keys to the Nematode Parasites of Vertebrates: Archival Volume; CABI: Wallingford, UK, 2009; ISBN 1-84593-572-1. [Google Scholar]
  30. Taylor, M.A.; Coop, R.; Wall, R.L. Veterinary Parasitology; John Wiley & Sons: Hoboken, NJ, USA, 2015; ISBN 1-119-07369-3. [Google Scholar]
  31. Torres, J.; Segovia, J.; Miquel, J.; Feliu, C.; Llaneza, L.; Petrucci-Fonseca, F. Helmintofauna Del Lobo Ibérico (Canis lupus signatus Cabrera, 1907). Aspectos Potencialmente Útiles En Mastozoologia. Galemys 2000, 12, 1–11. [Google Scholar]
  32. Čabanová, V.; Guimaraes, N.; Hurníková, Z.; Chovancová, G.; Urban, P.; Miterpáková, M. Endoparasites of the Grey Wolf (Canis lupus) in Protected Areas of Slovakia. Anna. Parasitol. 2017, 63, 283–289. [Google Scholar]
  33. Hermosilla, C.; Kleinertz, S.; Silva, L.M.; Hirzmann, J.; Huber, D.; Kusak, J.; Taubert, A. Protozoan and Helminth Parasite Fauna of Free-Living Croatian Wild Wolves (Canis lupus) Analyzed by Scat Collection. Vet. Parasitol. 2017, 233, 14–19. [Google Scholar] [CrossRef]
  34. De Liberato, C.; Grifoni, G.; Lorenzetti, R.; Meoli, R.; Cocumelli, C.; Mastromattei, A.; Scholl, F.; Rombolà, P.; Calderini, P.; Bruni, G.; et al. Angiostrongylus vasorum in Wolves in Italy: Prevalence and Pathological Findings. Parasites Vectors 2017, 10, 386. [Google Scholar] [CrossRef] [Green Version]
  35. Bagrade, G.; Kirjusina, M.; Vismanis, K.; Ozolins, J. Helminth Parasites of the Wolf Canis Lupus from Latvia. J. Helminthol. 2009, 83, 63. [Google Scholar] [CrossRef]
  36. Shimalov, V.; Shimalov, V. Helminth Fauna of the Wolf (Canis lupus Linnaeus, 1758) in Belorussian Polesie. Parasitol. Res. 2000, 86, 163–164. [Google Scholar] [CrossRef]
  37. Humm, K.; Adamantos, S. Is Evaluation of a Faecal Smear a Useful Technique in the Diagnosis of Canine Pulmonary Angiostrongylosis? J. Small Anim. Pract. 2010, 51, 200–203. [Google Scholar] [CrossRef]
  38. Schnyder, M.; Tanner, I.; Webster, P.; Barutzki, D.; Deplazes, P. An ELISA for Sensitive and Specific Detection of Circulating Antigen of Angiostrongylus vasorum in Serum Samples of Naturally and Experimentally Infected Dogs. Vet. Parasitol. 2011, 179, 152–158. [Google Scholar] [CrossRef] [PubMed]
  39. Jefferies, R.; Morgan, E.R.; Helm, J.; Robinson, M.; Shaw, S.E. Improved Detection of Canine Angiostrongylus vasorum Infection Using Real-Time PCR and Indirect ELISA. Parasitol. Res. 2011, 109, 1577–1583. [Google Scholar] [CrossRef] [PubMed]
  40. Houpin, E.; McCarthy, G.; Ferrand, M.; Waal, T.D.; O’Neill, E.J.; Zintl, A. Comparison of Three Methods for the Detection of Angiostrongylus vasorum in the Final Host. Vet. Parasitol. 2016, 220, 54–58. [Google Scholar] [CrossRef]
  41. Elsheikha, H.M.; Holmes, S.A.; Wright, I.; Morgan, E.R.; Lacher, D.W. Recent Advances in the Epidemiology, Clinical and Diagnostic Features, and Control of Canine Cardio-Pulmonary Angiostrongylosis. Vet. Res. 2014, 45, 92. [Google Scholar] [CrossRef]
  42. Gortázar, C.; Villafuerte, R.; Lucientes, J.; Fernández-de-Luco, D. Habitat Related Differences in Helminth Parasites of Red Foxes in the Ebro Valley. Vet. Parasitol. 1998, 80, 75–81. [Google Scholar] [CrossRef]
  43. Segovia, J.; Torres, J.; Miquel, J. Helminth Parasites of the Red Fox [Vulpes vulpes L., 1758] in the Iberian Peninsula: An Ecological Study. Acta Parasitol. 2004, 49, 67–79. [Google Scholar]
  44. Mañas, S. Cardiopulmonary Helminth Parasites of Red Foxes (Vulpes vulpes) in Catalonia, Northeastern Spain. Vet. J. 2005, 169, 118–120. [Google Scholar] [CrossRef]
  45. Gerrikagoitia, X.; Barral, M.; Juste, R. Angiostrongylus Species in Wild Carnivores in the Iberian Peninsula. Vet. Parasitol. 2010, 174, 175–180. [Google Scholar] [CrossRef]
  46. Torres, J.; Miquel, J.; Motjé, M. Helminth Parasites of the Eurasian Badger (Meles meles L.) in Spain: A Biogeographic Approach. Parasitol. Res. 2001, 87, 259–263. [Google Scholar] [CrossRef]
  47. Gillis-Germitshc, N.; Kapel, C.M.O.; Thamsborg, S.M.; Deplazes, P.; Schnyder, M. Host-Specific Serological Response to Angiostrongylus vasorum Infection in Red Foxes (Vulpes vulpes): Implications for Parasite Epidemiology. Parasitology 2017, 144, 1144–1153. [Google Scholar] [CrossRef] [PubMed]
  48. Morgan, E.R.; Shaw, S.E.; Brennan, S.F.; Waal, T.D.D.; Jones, B.R.; Mulcahy, G. Angiostrongylus Vasorum: A Real Heartbreaker. Trends Parasitol. 2005, 21, 49–51. [Google Scholar] [CrossRef] [PubMed]
  49. Morgan, E.R.; Jefferies, R.; Krajewski, M.; Ward, P.; Shaw, S.E. Canine Pulmonary Angiostrongylosis: The Influence of Climate on Parasite Distribution. Parasitol. Int. 2009, 58, 406–410. [Google Scholar] [CrossRef] [PubMed]
  50. Martínez-Carrasco, C.; Berriatua, E.; Garijo, M.; Martínez, J.; Alonso, F.; Ruiz de Ybáñez, R. Epidemiological Study of Non-systemic Parasitism in Dogs in Southeast Mediterranean Spain Assessed by Coprological and Post-mortem Examination. Zoonoses Public health 2007, 54, 195–203. [Google Scholar] [CrossRef] [PubMed]
  51. Colella, V.; Mutafchiev, Y.; Cavalera, M.A.; Giannelli, A.; Lia, R.P.; Dantas-Torres, F.; Otranto, D. Development of Crenosoma vulpis in the Common Garden Snail Cornu Aspersum: Implications for Epidemiological Studies. Parasites Vectors 2016, 9, 208. [Google Scholar] [CrossRef]
  52. Jeffery, R.A.; Lankester, M.W.; McGrath, M.J.; Whitney, H.G. Angiostrongylus vasorum and Crenosoma vulpis in Red Foxes (Vulpes vulpes) in Newfoundland, Canada. Can. J. Zool. 2004, 82, 66–74. [Google Scholar] [CrossRef]
  53. Miró, G.; Montoya, A.; Jiménez, S.; Frisuelos, C.; Mateo, M.; Fuentes, I. Prevalence of Antibodies to Toxoplasma gondii and Intestinal Parasites in Stray, Farm and Household Cats in Spain. Vet. Parasitol. 2004, 126, 249–255. [Google Scholar] [CrossRef]
  54. Muñoz, S.; Ramos, P.L.; Carretón, E.; Diosdado, A.; González-Miguel, J.; Simón, F.; Morchón, R. Intestinal helminths in Iberian wolves (Canis lupus signatus) from Northwest Spain. The Open Parasitology J. 2018, 6, 106–111. [Google Scholar] [CrossRef]
  55. Martínez-Carrasco, C.; Ruiz de Ybanez, M.; Sagarminaga, J.; Garijo, M.; Moreno, F.; Acosta, I.; Hernandez, S.; Alonso, F. Parasites of the Red Fox (Vulpes vulpes Linnaeus, 1758) in Murcia, Southeast Spain. Rev. Med. Vet. 2007, 158, 331–335. [Google Scholar]
  56. Criado-Fornelio, A.; Gutierrez-Garcia, L.; Rodriguez-Caabeiro, F.; Reus-Garcia, E.; Roldan-Soriano, M.; Diaz-Sanchez, M. A Parasitological Survey of Wild Red Foxes (Vulpes vulpes) from the Province of Guadalajara, Spain. Vet. Parasitol. 2000, 92, 245–251. [Google Scholar] [CrossRef]
  57. Rutgers, M.; Orgiazzi, A.; Gardi, C.; Römbke, J.; Jänsch, S.; Keith, A.M.; Neilson, R.; Boag, B.; Schmidt, O.; Murchie, A.K. Mapping Earthworm Communities in Europe. Appl. Soil Ecol. 2016, 97, 98–111. [Google Scholar] [CrossRef]
  58. Neubert, E.; Seddon, E.; Allen, M.; Backeljau, T. European Red List of Terrestrial Molluscs: Snails, Slugs, and Semi-Slugs; IUCN: Brussels, Belgium, 2019. [Google Scholar]
  59. Jefferies, R.; Shaw, S.; Willesen, J.; Viney, M.; Morgan, E. Elucidating the Spread of the Emerging Canid Nematode Angiostrongylus vasorum between Palaearctic and Nearctic Ecozones. Infect. Genet. Evol. 2010, 10, 561–568. [Google Scholar] [CrossRef] [PubMed]
  60. Millán, J.; García, E.J.; Oleaga, Á.; López-Bao, J.V.; Llaneza, L.; Palacios, V.; Candela, M.G.; Cevidanes, A.; Rodríguez, A.; León-Vizcaíno, L. Using a Top Predator as a Sentinel for Environmental Contamination with Pathogenic Bacteria: The Iberian Wolf and Leptospires. Memórias do Inst. Oswaldo Cruz 2014, 109, 1041–1044. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Animals 12 02289 g001 550
Figure 1. Cardiopulmonary nematode worms detected in wolves inhabiting the Galicia region (NW Spain).
Figure 1. Cardiopulmonary nematode worms detected in wolves inhabiting the Galicia region (NW Spain).
Animals 12 02289 g001
Table
Table 1. Numbers of cardiorespiratory nematodes collected from 14 infected Iberian wolves from northwestern Spain according to the species identified.
Table 1. Numbers of cardiorespiratory nematodes collected from 14 infected Iberian wolves from northwestern Spain according to the species identified.
Species IdentifiedNo. Positive Wolves (%)No. Total WormsFemale WormsMale WormsMean (SD)No. Worms per Wolf (min.–max.)
A. vasorum11 (19.2)374 *2657134 (35.1)1–99
C. vulpis4 (7)8712 (0.8)1–3
E. aerophilus2 (3.5)3301.5 (0.7)1–2
A. vasorum single infection8 (14)348 *2456543.5 (36.7)7–99
C. vulpis single infection3 (5.3)6512 (1.0)1–3
A. vasorum + C. vulpis1 (1.7)1813518 (0.0)18
A. vasorum + E. aerophilus2 (3.5)13 12 1 6.5 (6.4)2–11
* The sex of 38 worms of A. vasorum could not be determined: No. = number; Min. = minimum; Max. = Maximum.
Table
Table 2. Prevalence of Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus according to epidemiological category.
Table 2. Prevalence of Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus according to epidemiological category.
CategoryTotal No. WolvesAngiostrongylus vasorumCrenosoma vulpisEucoleus aerophilusTotal No. of Worms
No. Positive Wolves%p-ValueNo. Positive Wolves%p-ValueNo. Positive Wolves%p-ValueNo. Positive Wolves%p-Value
ProvinceA Coruna153200.959213.30.26500.01426.71
Lugo21523.800.014.8523.8
Ourense15213.3213.316.7426.7
Pontevedra6116.700.000116.7
SexMale29517.20.74726.9100.00.236724.11
Female28621.427.127.1725.0
Body condition (1–5)1200.00.22100.00.44400.0100.00.174
210440.0220.000.0550.0
320525.015.015.0630.0
42328.714.314.3313.0
5100.000.000.000.0
AgePup14321.41428.60.00217.10.714642.90.243
Young24520.800.014.1528.8
Adult19315.800.000.0315.8
Table
Table 3. Number of cardiorespiratory worms found per wolf according to host age.
Table 3. Number of cardiorespiratory worms found per wolf according to host age.
Wolf Pup (n = 14)Young (n = 24)Adults (n = 19)
No. of ParasitesNo.%p-ValueNo.%p-ValueNo.%p-Value
0857.10.0831979.20.7571684.20.343
1–29642.90.00428.30.27715.260.246
30-60000.56728.30.56615.261.000
>60001.00014.21.00015.261.000
No. = number.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Estévez-Sánchez, E.; Checa, R.; Montoya, A.; Barrera, J.P.; López-Beceiro, A.M.; Fidalgo, L.E.; Miró, G. A High Prevalence of Cardiopulmonary Worms Detected in the Iberian Wolf (Canis lupus): A Threat for Wild and Domestic Canids. Animals 2022, 12, 2289. https://doi.org/10.3390/ani12172289

AMA Style

Estévez-Sánchez E, Checa R, Montoya A, Barrera JP, López-Beceiro AM, Fidalgo LE, Miró G. A High Prevalence of Cardiopulmonary Worms Detected in the Iberian Wolf (Canis lupus): A Threat for Wild and Domestic Canids. Animals. 2022; 12(17):2289. https://doi.org/10.3390/ani12172289

Chicago/Turabian Style

Estévez-Sánchez, Efrén, Rocío Checa, Ana Montoya, Juan Pedro Barrera, Ana María López-Beceiro, Luis Eusebio Fidalgo, and Guadalupe Miró. 2022. "A High Prevalence of Cardiopulmonary Worms Detected in the Iberian Wolf (Canis lupus): A Threat for Wild and Domestic Canids" Animals 12, no. 17: 2289. https://doi.org/10.3390/ani12172289

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop