Parasitology Research https://doi.org/10.1007/s00436-018-5984-6 SHORT COMMUNICATION
Man. num.: PARE-D-18-00358R3 https://rdcu.be/1uC2 Published online 23 June 2018
The first reported case of advanced aelurostrongylosis in Eurasian badger (Meles meles, L.1758) in Bosnia and Herzegovina: histopathological and parasitological findings Oliver Stevanović1*, Igor Trbojević2, Sonja Nikolić1and Violeta Santrač1 Public Veterinary Institute of the Republika Srpska „Dr. Vaso Butozan” Banja Luka, Branka Radičevića 18, 78000 Banja Luka, Bosnia and Herzegovina 2 Faculty of Science, University of Banja Luka, Mladena Stojanovića 2, 78000 Banja Luka, Bosnia and Herzegovina; ORCID: orcid.org/0000-0003-0781-7235 *Corresponding author: Mobile: +38765707318; E-mail:
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Abstract This paper represents the first description of advanced aelurostrongylosis in the Eurasian badger (Меles meles L. 1758) from Bosnia and Herzegovina, which is an autochthonous species of the country. An adult female badger was found dead on a road; the cause of death was trauma but the emphasis in the paper is on severe verminous pneumonia caused by metastrongylids from genus Aelurostrongylus spp. This parasitological and histopathological finding confirms the presence of Aelurostrongylus in mustelids in Bosnia and Herzegovina and the Balkans. Keywords: Aelurostrongylosis, Bosnia and Herzegovina, Eurasian badger, histopathology, Meles meles
The Eurasian badger is an autochthonous, hunted and unprotected species of Bosnia and Herzegovina (Official Gazette of the Republic of Srpska, 2009). The badger’s meat is used for human consumption in some areas of the Balkans (Konjević, 2005), and more importantly, it is locally believed that the badger’s fat has certain beneficial medical effects. Schlegel (1933) described the lungworm disease as a health problem in the Eurasian badger. Until today, there are several reports on pulmonary nematodes in Eurasian badgers. The following two species of genus Aelurostrongylus have been reported in the European badger: Aelurostrongylus pridhami (Anderson, 1962) in Spain (Torres et al. 2001) and Aelurostrongylus falciformis (Schlegel 1933) in the Russian Federation (Kontrimavichus et al. 1976), Italy (Magi et al. 1999), Germany (Stube 1965), England (Jones et al. 1980), Norway (Davidson et al. 2006) and Poland (Demiaszkiewicz et al. 2017). Apart from A. pridhami and A. falciformis, other lungworms found in badgers include the following metastrongyles: Angiostrongylus vasorum, Angiostrongylus daskalovi, Crenosoma melesi, Crenosoma vulpis and trichuroid Capillaria aerophila (Davidson et al. 2006; Popiołek et al. 2009). In the Balkans, there are certain data on presence of intestinal helminths in badgers from Serbia (Bošković and Valter 1979), but pulmonary nematodes were not recorded. Additionally, there are no published data on parasitic diversity in badgers in Bosnia and Herzegovina. The mentioned papers mostly describe Aelurostrongylus spp. in badgers, whereas the clinical significance and impact of the parasite on badgers’ health require far more investigation. This paper describes the first report of aelurostrongylosis in badgers with severe pulmonary lesions – verminous pneumonia.
An adult female badger was found dead on a road (N 44º41'1.91"; E 17º05'1.75") in the Mount Manjača region (near Banja Luka), suggesting a typical road kill. The badger’s carcass was collected and kept in a freezer at -20 ºC until parasitological post mortem examination. The necropsy was performed in the Pathology Laboratory of the Public Veterinary Institute „Dr. Vaso Butozan“ in Banja Luka. The badger was in good body condition, its weight being 14.9 kg. and body length was 65 cm without tail. The cause of death was trauma, which was established by external laceration of skin, subcutaneous hematomas, obvious rupture of the abdomen, presence of gastric contents in the abdominal cavity, and congestion of the spleen.
Fig. 1. a) Aelurostrongylusin lung parenchyma–in situ; b) Verminous pneumonia– histopathology Macroscopical changes in the lungs were evident. They were firm, grey-yellowish, disseminated, solid nodules of various sizes, which tended to converge. After dissection of lungs (Figure 1a), tiny white pulmonary nematodes were found in the terminal bronchial airways and parenchyma. Most notably, a huge number of worms was found in the diaphragmatic lobes. Morphologically-altered tissue samples from lungs were fixed in 10% formalin, embedded in paraffin, cut into 4 µm sections and stained with hematoxylin and eosin (HE). Histologically, severe verminous pneumonia, with a large number of adult pulmonary nematodes with larvae and eggs, was observed (Fig 1b). The nematodes were visible in alveoli and terminal bronchial pathways with a clearly expressed reaction of the tissue surrounding the parasites. In some cases, alveolar walls were ruptured. Interalveolar septae were thicker due to infiltration of leukocytes, with the highest proportion of mononuclear and eosinophil cells. The connective tissue suffered from hypertrophy and hyperplasia of the smooth-muscular layer of terminal bronchioles and alveolar channels, which was similar to observations by Gavrilović et al. (2017). The final parasitological diagnosis up to genus was made upon the morphological characteristics of male and female nematodes and larvae found in lungs, according to descriptions made by Anderson (1962); Kontrimavichus (1985); Davidson et al. (2006) and Kretschmar, (2016). Dissected lungs were placed in a deep tray and rinsed under the jet of tap water and left several minutes to sediment. The visible worms in sediment were transferred to Petri dishes filled with 0.9% NaCl solution. Some nematodes were detached from lung tissue with a pair of tweezers. The worms (whole and fragmented ones) were cleared with Amann’s lactophenol and observed under a light microscope (x40, x100, x400). At the anterior end of nematodes, a small buccal opening with cephalic papillae, teguminal sheath, and thin cuticle was visible. In females, easily visible uterus filled with larvae could be observed (Fig 2a). In males, the most important diagnostic feature was the small and narrow copulatory bursa with short rays, and short, relatively thick spicules with well-developed chitinous gubernaculum (from lateral projection, gubernaculum is visible as a narrow additional piece of spicules).
Fig. 2. a) Uterus filled with larvae in a female of Aelurostrongylus; b) Aelurostrongylus male copulatory bursa The spicules were equal in length and curved ventrally. The proximal nodular tips of spicules were visibly thicker than the distal part, which ended in a solid, slightly nodular point (Figure 2b). In the uterus of the females, L1 with a wavy tail and dorsal notch were observed. Davidson et al. (2006) and Demiaszkiewicz et al. (2017) made similar observations. Some morphological details (structure and position of bursal rays) from basic descriptions (Wetzel 1938; Anderson 1962; Kontrimavichus 1985) were not observed in this case. However, the morphology of gubernaculum and spicules indicated that the species was most probably Aelurostrongylus falciformis (Schegel, 1933). Detailed morphometrical study of worms, necessary for species diagnosis was not performed in this study. Therefore, the diagnosis was concluded to the genus level – Aelurostrongylus spp. based on the following facts: location of the parasites in lung tissue, the general morphology, the morphology of copulatory bursa, male spicules and gubernaculum, and the morphology of L1 with a wavy tail and notch in the uterus of the females. This paper presents the first report on genus Aelurostrongylus spp. in mustelids in Bosnia and Herzegovina and the Balkans, which implies that these nematodes (considering their complex development cycle) are widespread among wildlife. Given that there is limited data on the genus Aelurostrongylus in the literature on mustelids, it is necessary to thoroughly examine the distribution of species A. pridhami and A. falciformis based on detailed morphological and molecular studies, as in the case of Angiostrongylus (Gherman et al. 2016). In the latest German study (Kretschmar, 2016), the morphological differences between A. pridhami and A. falciformis were reviewed. The author describes in detail all morphological structures (formation of copulatory bursa, morphometry of spicules, shape and size of gubernaculum) from the basic literature, which are important in the diagnosis of species (Wetzel 1938; Anderson 1962; Kontrimavichus 1985). In case of the examined Aelurostrongylus spp. from Mustela putorius (L., 1758), certain morphological characteristics were unclear, regarding the two mentioned species that are common in mustelids. Consequently, Kretschmar (2016) did not establish the specific diagnosis. The described advanced lesions in the lungs indicate that metastrongyloid species from genus Aelurostrongylus can be a serious pathogen in badgers, similar to Aelurostrongylus abstrusus in felids (Grabarević et al. 1999; Gavrilović et al. 2017). However, it is impossible to measure the impact of Aelurostrongylus on the badger’s health, without knowing the condition and weight of the examined animal's body. In addition, this report represents a case study, so it is not possible to fully describe the impact of Aelurostrongylus on badgers without conducting further clinical trial that includes a larger sample size and a detailed epidemiological study. The histopathological lesions in the lung due to an enormous number of worms indicate chronic and advanced process, which is suggestive for possible expression of clinical signs during life.
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