3 Contribution for the knowledge of Wild Boar (Sus scrofa L.) helmintic fauna in Tapada Nacional de Mafra, an enclosured hunting area.

Bruno de Sousa, C.1; de Carvalho, L.M.2; Fazendeiro, I.2; Castro Rego, F.3; Afonso-Roque, M.M.1 1 Instituto Investigação Científica Tropical - Centro de Veterinária e Zootecnia, Lisboa, . 2 Faculdade de Medicina Veterinária - Parasitologia e Patologia das Doenças Parasitárias, Lisboa, Portugal. 3 Centro de Ecologia Aplicada Prof. Baeta Neves - Intituto Superior de Agronomia, Lisboa, Portugal.

Received: 29.10.03 Accepted: 01.05.04

Abstract: Aiming a better knowledge of wild boars helmintic fauna in Portugal, the study of adult helminths was performed in samples from animals hunted during the hunting season of 2000 and 2001 (April - July) at Tapada Nacional de Mafra. Liver (n=23) and lungs (n=19) were available in the two hunting seasons, faeces only in 2000 and gastrointestinal tracts were examined only in 2001. Fasciola hepatica Linnaeus, 1758 and Cysticercus tenuicollis were found in 60,8% and 4,3% of the livers (N=23). Lungworms were collected from 42% of the hosts. Metastrongylus apri Gmelin, 1790, Vostokov, 1905 and M. salmi Gedoelst, 1923 were found in association. M. salmi is identified for the first time in this host, in Portugal. Globocephalus urosubulatus Alessandrini, 1909, Oesophagostomum dentatum Rudolphi, 1803, Molin, 1861, Ascarops strongylina Rudolphi, 1819 and Trichuris suis Schrank, 1788 were collected from the gastrointestinal contents (N=8) of 62,5%, 25%, 12,5% and 12,5%, respectively. Keywords: Wild boar, gastrointestinal worms, lungworms, Portugal.

Resumen: El presente estudio busca contribuir al conocimiento de la helmintofauna del jabalí en Portugal. Para ello han sido estudiadas muestras provenientes de jabalíes abatidos en la Tapada Nacional de Mafra durante las épocas de caza correspondientes a los años 2000 y 2001 (Abril-Julio). Los hígados (N=23) y los pulmones (N=19) han estado a disposición durante en los 2 periodos de caza, las heces (N=6) en 2000 y los tractos gastrointestinales (N=8) solamente han sido observados en el año 2001. Fasciola hepatica Linnaeus, 1758 y Cysticercus tenuicollis han revelado prevalencias del 60,8% y 4,3%, respectivamente en los hígados observados. Metastrongylus apri Gmelin, 1790, Vostokov, 1905 y M. salmi Gedoelst, 1923 fueron observados asociadamente en el 42% de los hospedadores. M. salmi se detecta por primera vez en Portugal. Los parásitos hallados en el tracto gastrointestinal, junto a sus prevalencias, han sido los siguientes: Globocephalus urosubulatus Alessandrini, 1909 (62,5%); Oesophagostomum dentatum Rudolphi, 1803, Molin, 1861 (25%); Ascarops strongylina Rudolphi, 1819 (12,5%) y Trichuris suis Schrank, 1788 (12,5%).Palabras clave: protozoos intestinales, quistes, trofozoitos, edad, sexo, humanos, Libia. Palabras Clave: Jabalí, helmintos gastrointestinales, parásitos pulmonares, Portugal.

1. Introduction can be relevant in the natural reserves / parks and The study of risk factors for the maintenance game areas. of wildlife health status, such as parasitic diseases, In wild populations there is usually a balanced host-parasite relationship, not compromising animal health and welfare. However, anthropogenic changes

Corresponding author: of the environment, the increase of human populations Dra. Carolina Bruno de Sousa and the introduction of other animal species, may Address: Instituto Investigação Científica Tropical - Centro de Veterinária e Zootecnia, Pólo Universitário Alto da Ajuda, introduce stress factors that can disrupt the natural Rua Prof. Cid dos Santos 1300-477 Lisboa, Portugal. balance and induce pathological situations. This study Tel: +351213652800, ext 1323 is a contribution for the knowledge of wild boar Fax: +351213652869 e-mail: [email protected] helmintic fauna and for a better understanding of the

Revista Ibérica de Parasitología (2004), 64 (1-4), 3-7. © 2004 Sociedad Española de Parasitología (SEP) 4 Bruno de Sousa, C., et al., Wild Boar (Sus scrofa L.) helmintic fauna. host/parasite relationship in this field of animal Biometrical parameters (body, kidney and parasitology in Portugal, since there are not many spleen weights, percentage of kidney fat and studies about this subject in our country (Barata, 1989; corporal lengths – total length, ear, tail, hock and Caeiro et al., 1993, Bruno de Sousa et al., 2000, 2001). garrote), age and sex were also determined and correlated with parasitological parameters (EPG, 2. Material and Methods FE, prevalence of lungworms in lungs and the Tapada Nacional de Mafra is a national hunt intensity of F. hepatica in the liver) of the 2000 and leisure park since 1747. This limited area with hunting season animals. ANOVA analysis was done 819 ha is located 30 Km North of , included through Spearman matrix in view to overtake the in an irregular orographic region (80-358m). heterogeneity of the sample, followed by a principal During the 2000 and 2001 hunting seasons components analysis with the obtained matrix. (April-July), was possible to collect heterogeneous Descriptive statistics and statistical analysis samples from 23 adult wild boars (7 females and 16 were performed with Excel for Microsoft Office males) hunted at Tapada Nacional de Mafra. Livers 2000 and Statistica for Windows, version 6.0, using (n=23), lungs (n=19), gastrointestinal tracts (n=8) a significance level of p<0,01. and faeces (n=6) were stored at 4ºC before being processed (Table 1). 3. Results and Discussion Parasitic forms (helmintic adult forms and/or Table 1. Constitution of the studied sample eggs) were detected in 78,2% of the studied wild boars. All faecal samples were positive to the presence Date Animals Lungs Liver Gastrointestinal tractFaeces of gastrointestinal strongyles eggs revealing an average 1x x 2000 3 x EPG of 2142. A very important egg shedding, up to 6xx 3000 EPG, was evidence in 33% of the samples (Fig.5). 5xx x Joining EPG results with the presence of 2001 8 x x x nematode adult forms verified in 5 animals, was Total 23 19 23 8 6 possible to considerer a prevalence of 61,1% for the presence of gastrointestinal nematodes. Parasitic adult forms were collected from the The identification, prevalence and mean organs thought parasitological classical methods and intensity of the adult specimens collected from the submitted to morphological study (Bowman, 1995). gastrointestinal organs is showed in Table 2. The presence of Fasciola was also determined by Table 2. Prevalence and mean intensity of helminths in the stu- the presence of eggs in the bile per ml (FE). died population (N) Faecal egg counts (eggs per gram - EPG) were determined with McMaster slide using N Prevalence (%) Mean intensity Site location samples directly collected from the rectum Globocephalus urusobulatus 8 62,5 205,6 Small intestine (Thienpont et al., 1986). Ascarops strongylina 8 12,5 1 Small intestine According to Bush et al. (1997) two Trichuris suis 8 12,5 1 Large intestine Oesophagostomum dendatum 8 25,0 19 Caecum parameters were used to evaluate the infection Metastrongylus spp 19 42,1 ? Lungs level: prevalence (number of hosts infected with 1 Fasciola hepatica 23 60,9 14 Liver or more individuals of a particular parasite species ? - not countable divided by the number of hosts examined for that parasite species) and mean intensity (total number Globocephalus urosubulatus was found in of parasites of a particular species found in a sample the small intestine, showed the higher prevalence divided by the number of hosts infected with that (62,5%) and mean intensity (min. 13, max. 345) of parasite). the helminths found in the gastrointestinal tract Bruno de Sousa, C., et al., Wild Boar (Sus scrofa L.) helmintic fauna. 5

(Fig. 1). Oesophagostumum dentatum was present in the caecum of 2 animals (25%), with a maximum of 22 and a minimum of 16 specimens/host (Fig. 2). One specimen of Ascarops strongylina and other of Trichuris suis were found in 2 different animals (12,5%) (Fig. 3). Fasciola hepatica prevalence was 60,8%, and its mean intensity ranged between 1 and 41 specimens. In what concerns FE, the average was 821. However, one individual was highly infected, with an egg count reaching more than 8000 eggs/ml of bile. The FE/F.hepatica relation range between Fig 1. Globocephalus urosubulatos. Anterior end (A) and male poste- 2,4 and 1010 with an average of 105 FE/F.hepatica, rior end (B). Originals. but any significant correlation was found between these two parameters. To our knowledge, the absence of references to wild boar parasitism by Fasciola hepatica in other regions and countries, indicate that the situation here studied, and previous described by other Portuguese authors (Barata, 1989; Bruno de Sousa et al.. 2000, 2001) is unusual. At Tapada Nacional de Mafra, wild boar population shares his enclosured territory with a fallow deer population with a high prevalence of fasciolosis (Bruno de Sousa et al., 2001) fact that certainly conducted to a change in the host–parasite balance. However these results doesn’t allow Fig 2. Oesophagostomum dentatum. Anterior end (A) and male poste- advancing with more conclusions. rior end (B). Originals. Cysticercus tenuicollis was also found in the liver of one wild boar. Metastrongylus apri and M. salmi were found in association, in 42% of the hosts. Despite adult specimens were found in lungs airways, any lungworms eggs were found in the examined faeces, what could be related with the 4ºC faeces storage. M. salmi is here describe for the first time in this host in Portugal, although M. apri was previously recorded by Barata & Afonso-Roque (1992) (Fig. 4). Despite high value of the found prevalence, it is a half of the referred by de-la-Muela et al., 2001 to this genus at eastern , what remembers the high importance of this infection in this host All nematode findings are in agreement with the results obtained in other regions of the Iberian Peninsula (de-la-Muela et al., 2001 and Fernandez- Fig 3. Ascarops strongylina. Anterior end (A) and male posterior end de-la-Mera et al., 2003). (B). Originals. 6 Bruno de Sousa, C., et al., Wild Boar (Sus scrofa L.) helmintic fauna.

variation is explained by axis 2 and it seems to be more related with the age factor (Fig.5). Variance analysis suggests a significant correlation between female sex and Fasciola hepatica infection (p<0,0178), with an F (1,13)=7,35.

4. Conclusions Fig 4. Females posterior end of the species Metastrongylus apri (A) With exception to Fasciola hepatica, the and M. salmi (B). Originals. present results agree with other temperate regions, namely in what concerns the infection by lungworms Spearman analysis did not show significant and gastrointestinal nematodes (Chinone et al. 1984; correlations (p<0,01) between parasitological and Humbert & Henry, 1989; Takacs, 1996, 1997, de-la- biometrical parameters. However, principal Muela et al., 2001; Fernandez-de-la-Mera et al., components analysis of the Spearman correlations 2003). matrix data, concludes that axis 1 (explaining Concerning in detail the high levels found of 37,74% of the variation) with variables related with parasitic infections, Globocephalus urosubulatus weight and animal dimension, shows a positive and Fasciola hepatica showed the highest mean tendency with some parasitological variables (EPG, intensity and both are pathogenic to swine, being FE, lungworm infection). No relation was the second one a zoonosis. Oesophagostomum evidenced between Fasciola hepatica infection and dentatum was also present, being well known by its the other variables. Only 16,71% of the total pathogenic action in the large intestine.

Fig 5. Principal components analysis made with Spearman correlations matrix data of the biometrical and parasitological variables (factor 1 vs factor 2). Bruno de Sousa, C., et al., Wild Boar (Sus scrofa L.) helmintic fauna. 7

The average EPG showed high elimination Afonso-Roque, M.M. 2001. Fauna parasitária das levels by gastrointestinal nematodes, meaning that populações de gamo (Dama dama L.) e javali (Sus this kind of parasitc infection can be very common scrofa L.) da Tapada Nacional de Mafra. Acta in this host. Parasitológica Portuguesa, 8, 236. Relevant lungworm infection was detected, Bruno de Sousa, C.; Maia, M.J.; Afonso-Roque, M.M. e with a new recorded species, Metastrongylus salmi, Fazendeiro, M.I. 2000. Contribuição para o conhecimento for this host in Portugal. da fasciolose nas populações de gamo (Dama dama L.) e It is possible to conclude that the most javali (Sus scrofa L.) na Tapada Nacional de Mafra. Acta prevalent gastrointestinal parasites found are Parasitológica Portuguesa, 7, 98-99. pathogenic and the recorded faecal egg counts must Bush, A.O.; Lafferty, K.D.; Lotz, J.M. and Shostak, A.W. 1997. have a major contribution by these parasites. Parasitology meets ecology on its own terms: Margolis Accordingly, they must have more economic and et al. revisited. J Parasitol, 83, 575-583. sanitarian importance than our sample was able to Caeiro, V.; Pereira, M.A. and Vila Viçosa, M.J. 1993. show in both parasitological and biometrical Contribuição ao estudo dos endoparasitas do javali. correlations. Acta Parasitológica Portuguesa, 1, 15-19. Chinone, S.; Itagaki, H.; Hoshins, C. and Inoaka, H. 1984. Three 5. Acknowledgements species of Metastrongylus lungworms from a wild boar PRODEP - Portuguese Program for the Educative Development on Iriomote Island (Nematoda: Strongyloidea). Bulletin for research funding. To Eng. Ricardo Paiva, Eng. Pedro Carrilho of the Azabu University Veterinary Medicine, 5, 109-118. and Forest Guards, Mr. João Louro and Mr. Filipe Claro from De-la-Muela, N.; Hernández – de Lujan, S. and Ferre, I. 2001. Tapada Nacional de Mafra, for the field support. To Lídia Gomes Helminths of wild boar in Spain. J Wildlife Dis, 37, 840- from the Parasitology and Parasitic Diseases Laboratory, Faculty 843. of Veterinary Medicine of Lisbon for the laboratorial assistance. Fernandez-de-Mera, I.G.; Gortazar, C.; Vicente, J.; Hofle, U. and Fiero, Y. 2003. Wild boar helminths. Risks in 6. References animal translocations. Vet. Parasitol, 115, 335-341. Barata, M.C. e Afonso-Roque, M.M. 1992. As espécies do Humbert, J.F. and Henry, C. 1989. Studies on the prevalence género Metastrongylus Molin, 1861 (Nematoda and transmission of lung and stomach nematodes of the Trichostrongyloidea) parasitas do javali (Sus scrofa wild boar (Sus scrofa) in France. J Wildlife Dis, 25, 335- castilianus Thomas, 1912) em Portugal. Garcia de 341. Orta, Série Zoológica, 16, 45-50. Takacs, A. 1996. Endoparasite infections of wild pigs (Sus Barata, M.C. 1989. Sobre a Existência de Fasciola hepatica L., scrofa L.) in Hungary. Magyar Állatorvosok Lapja, 51, (1758), em animais Silvestres da Tapada Nacional de 721-724. Mafra - Estudo Preliminar. Garcia de Orta, Série Takacs, A. 1997. Contribution to the helminth fauna of wild Zoológica, 14, 29-33. boar (Sus scrofa) in Hungary. Wiener Tierarztliche Bowman, D.D. 1995. Georgi’s Parasitology for Veterinarians. Monatsschrift, 84, 314-316. (with a chapter on Antiparasitic Drugs by LYNN, R.C.). Thienpont, D.; Rochette, F. et Vanparijs, O.F.J. 1986. 6th edition, W.B. Saunders, Philadelphia, USA, 430 pp. Diagnóstico de las helminthiasis por medio del examen Bruno de Sousa, C.; Lourenço, A.M.; Madeira de Carvalho, coprológico. 2ª Edición, Janssen Research Foundation, L.M.; Pereira da Fonseca, I.M.; Fazendeiro, M.I. e Beerse, Belgium, 205 pp.