LITHUANIAN UNIVERSITY OF HEALTH SCIENCES VETERINARY ACADEMY Faculty of Veterinary Medicine Constantin Marc David Heinicke A relative study on association between Sarcoptes scabiei and intestinal helminth infection in a population of hunted red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in Lithuania. Asociacijos tarp užsikrėtimo Sarcoptes scabiei ir helmintais vertinimas Lietuvoje sumedžiotų rudųjų lapių (Vulpes vulpes) ir usūrinių šunų (Nyctereutes procyonoides) populiacijose. MASTER THESIS of Integrated Studies of Veterinary Medicine Supervisor Prof. Dr. Mindaugas Šarkūnas Kaunas, LT 2020 Annex 4 THE WORK WAS DONE IN THE DEPARTMENT OF VETERINARY PATHOBIOLOGY CONFIRMATION OF THE INDEPENDENCE OF DONE WORK I confirm that the presented Thesis “A relative study on association between Sarcoptes scabiei and intestinal helminth infection in a population of hunted red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in Lithuania.”. 1. Has been done by me; 2. Has not been used by any other Lithuanian or foreign university; 3. I have not used any other sources not indicated in the work and I present the complete list of the used literature. Constantin Marc David Heinicke (date) (name, surname) (signature) CONFIRMATION ABOUT RESPONSIBILITY FOR CORRECTNESS OF THE ENGLISH LANGUAGE IN THE DONE WORK I confirm the correctness of the English language in the done work. Genė Pečiukevičienė (date) (editor’s name, surname) (signature) CONCLUSION OF THE SUPERVISOR REGARDING DEFENCE OF THE THESIS Prof. Dr. Mindaugas Šarkūnas (date) (name, surname of the head of (signature) department/clinic/institute) Reviewer of the Master Thesis (name, surname) (signature) Evaluation of the defense commission of the Thesis: (date) (name, surname of the secretary of the (signature) defense commission) 2 | P a g e 1. TABLE OF CONTENT 1. TABLE OF CONTENT _______________________________________________________ 3 2. TABLE OF FIGURES AND IMAGES ___________________________________________ 4 3. SUMMARY ________________________________________________________________ 5 4. SANTRAUKA ______________________________________________________________ 7 5. INTRODUCTION ____________________________________________________________ 9 6. LITERATURE REVIEW _____________________________________________________ 12 6.1. Red fox (Vulpes vulpes) ___________________________________________________ 12 6.2. Raccoon dog (Nyctereutes procyonoides) _____________________________________ 13 6.3. Synergism or Mutualism of parasitic species __________________________________ 13 6.4. Coprological analysis of endoparasites _______________________________________ 14 6.5. Red fox and raccoon dog endoparasites_______________________________________ 14 6.5.1. Sarcoptes scabiei ____________________________________________________ 14 6.5.2. Nematodes _________________________________________________________ 15 6.5.3. Cestodes ___________________________________________________________ 17 6.5.4. Trematodes _________________________________________________________ 17 6.5.5. Protozoa ___________________________________________________________ 18 7. METHODS AND MATERIALS _______________________________________________ 19 7.1. Biosecurity _____________________________________________________________ 19 7.2. Collection of sample individuals ____________________________________________ 19 7.3. Identification and tracking of samples ________________________________________ 20 7.4. Test group _____________________________________________________________ 20 7.5. Coprological analyses ____________________________________________________ 22 7.5.1. Sample preparation ___________________________________________________ 22 7.5.2. Sample analysis _____________________________________________________ 22 7.6. Statistical analysis _______________________________________________________ 23 8. RESEARCH RESULTS ______________________________________________________ 25 9. DISCUSSION ______________________________________________________________ 34 10. CONCLUSION _____________________________________________________________ 37 11. PERSONAL SUGGESTION __________________________________________________ 38 12. BIBLIOGRAPHY ___________________________________________________________ 39 3 | P a g e 2. TABLE OF FIGURES AND IMAGES Figure 1 – The distribution of hunted and collected red fox test subjects in different sex and age groups, the title and number indicate the individuals in each group. ................................................ 21 Figure 2 – The distribution of hunted and sampled raccoon dog test subjects in different sex and age groups, the number indicating the individuals in each group. ........................................................... 22 Figure 3 – Prevalence of Sarcoptes sp. in hunted Raccoon dog (light grey) and red fox (dark grey). Numbers represent species prevalence in percentage........................................................................ 25 Figure 4 – Local distribution of Sarcoptic mange in percent of all gathered samples in Lithuanian provinces. ........................................................................................................................................... 26 Figure 5 – The detection frequency of excreted helminth eggs and coccidian oocysts in hunted Red fox and Raccoon dog sample population. .......................................................................................... 27 Figure 6 – The frequency of Sarcoptes scabiei and a selected parasite with the calculated prevalence for each species in Red fox or Raccoon dog. The “*“showing statistical significance of (p<0, 05) and “**” showing high statistical significance of (p<0.005)............................................................. 29 Figure 7 – Comparison of cross detection of red foxes which have Sarcoptes scabiei and a specific other parasite. The “*” shows a p value that is below 0, 05 and “**” shows a high statistically significant with (P < 0.005). The “^” suggest synergistic or mutualistic activity between parasites. The “˅“suggests that they have a negative effect on each other. ...................................................... 31 4 | P a g e 3. SUMMARY The presented thesis with the Title: “A relative study on association between Sarcoptes scabiei and intestinal helminth infection in a population of hunted red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in Lithuania.” was completed by me, Constantin Marc David Heinicke. To date, there is a lack of published information available on Sarcoptes scabiei and their interaction with other parasite species. In present study the association between ectoparasite Sarcoptes scabiei and detected intestinal parasite infections in hunted red foxes and raccoon dogs was evaluated, which gives an estimative view on prevalence of parasite infections in Lithuania’s medium sized carnivore populations as well as the relation between parasite species found and their abundance is highlighted in the present study. All foxes and raccoon dogs were divided into two groups each. The foxes in the first group (n =27) were infected with Taeniidae spp; Toxocara canis, Toxascaris leonina, Uncinaria stenocephala, Eucoleus aerophila, Capillaria putorii, Trichuris vulpis, Alaria alata, Coccidia spp; while foxes in the second group (n= 13) were infected with Sarcoptes sp. and one or more of the following: Toxocara canis, Uncinaria stenocephala, Capillaria putorii, Trichuris vulpis, Alaria alata, Coccidia spp.. The raccoons in the third group (n=6) were non-infected with Sarcoptes scabiei but these parasites were found: Toxocara canis, Toxascaris leonina, Uncinaria stenocephala, Eucoleus aerophila, Capillaria putorii, Trichuris vulpis, Alaria alata. While in the fourth group (n=8) raccoon dogs were infected with Sarcoptes sp. and one or more of each of these intestinal parasites: Toxocara canis, Uncinaria stenocephala, Capillaria putorii, Trichuris vulpis, Alaria alata. All foxes and raccoon dogs analyzed in present study were feral and had been obtained from all, but three provinces (Alytaus, Marijampoles and Telšiai) of Lithuania. All age groups and both sexes were gathered all year round. In total 88 animals were coprologically examined. The study was carried out from December 2017 till May 2020, at the Department of Veterinary Pathobiology, Veterinary Academy, LUHS. The parasite eggs which were found in the excrements of the test groups show a correlation between Sarcoptes sp. and endoparasites in both populations. As a matter of fact, we have the data suggesting the mutualistic or synergistic behavior as well as the reverse effect in which, for instance Uncinaria stenocephala showed a 4.40% increase (Ρ = 0.020) in raccoon dog population and a 3.80% (Ρ = 0.013) increase in the red fox population, the prevalence observed with the combination of Sarcoptes scabiei in contrast when Uncinaria stenocephala appeared on its own. The mechanisms causing it are still not known. For more detailed analysis, the investigation on this topic should be continued and more factors, for 5 | P a g e example host immunity parameters, parasitic behavior regarding the hosts temperature change, the behavioral changes of the host due to Sarcoptosis and its effects on alternate parasite infection. Larger sample sizes of feces should be collected as well. Keywords: Sarcoptes scabiei, coinfection – disease, endoparasites, Vulpes vulpes, Nyctereutes procyonoides, Lithuania; 6 | P a g e 4. SANTRAUKA Aš, Constantin Marc David Heinicke,