doi: 10.15389/agrobiology.2020.2.378eng
UDC: 636.934.57:619:616.993.192.1-07
Acknowledgements:
The authors thank colleagues from the Yakimov Department of Parasitology for help in identifying eimeriids and translation, the staff of the Department of Biology, Ecology, Histology (St. Petersburg State Academy of Veterinary Medicine) and TLVet Path International Consultants (Animal Eye Consultants of Iowa Laboratory, USA) for their assistance in tissue section preparation, histological pathomorphological analysis, and INC tests, and also we thank M.R. Kabilov (Gemomics SB RAS, Novosibirsk) for conducting molecular analyzes
PECULIARITIES OF DIAGNOSTICS AND PATHOMORPHOLOGY OF EIMERIIDOSES IN THE MINK FARMS OF THE NORTHWESTERN REGION OF THE RUSSIAN FEDERATION
Yu.E. Kuznetsov1, L.M. Belova1, N.A. Gavrilova1, M.E. Mkrtchyan1, K.V. Sidorenko1, А.B. Muromtsev2
1Saint-Petersburg State Veterinary Academy, 5, ul. Chernigovskaya, St. Petersburg, 196084 Russia, e-mail Fish2017@yandex.ru (✉ corresponding author), larissabelova2010@yandex.ru, nadezhda.gavrilova65@mail.ru, laulilitik@yandex.ru, capricorn26.12.94@yandex.ru;
2Kaliningrad State Technical University, 1, Sovetskii prosp., Kaliningrad Province, Kaliningrad, 236022 Russia, e-mail muromtsev.a@mail.ru
ORCID:
Kuznetsov Yu.E. orcid.org/0000-0001-9095-7049
Gavrilova N.A. orcid.org/0000-0001-5651-5976
Belova L.M. orcid.org/0000-0003-4473-1940
Sidorenko K.V. orcid.org/0000-0001-8987-9426
Mkrtchyan M.E. orcid.org/0000-0002-0615-5703
Muromtsev А.B. orcid.org/0000-0002-2456-7023
Received September 12, 2019
Parasitic diseases are widespread in fur-bearing animals, especially in minks. Coccidiidoses occupy a special place among invasive diseases, as they often occur without any symptoms and in some cases are not timely diagnosed. Despite the mild clinical manifestation of invasion, it causes serious damage to animal health and significant economic damage to fur-bearing animal farms. The pathogenic effect of eimeriids on the body of fur-bearing animals consists of mechanical, toxic and inoculative effects. As a result, accumulations of mucus are found in the intestinal contents, sometimes with bloody patches. Subacute catarrhal hemorrhagic enteritis occurs, which is manifested by areas of hyperemia and edema of the mucous membrane of the small intestine, desquamation of the epithelium and is accompanied by a violation of the structure of the villi. In the presented work, in the fur bearing animal farms of the Northwestern region of the Russian Federation, the parasitic fauna, prevalence rates (PR) and invasion intensity (II) of minks were studied for the first time, the species composition of eimeriids was clarified by the molecular-genetic method, the clinical and biochemical composition of blood, as well as pathomorphological changes in intestines in animals with eimeriidosis were studied. Isospora eversmanni was discovered in the Kaliningrad region for the first time and we managed to discover ill mink puppies since 13 days of their age. Even with low II in adult minks, pathomorphological changes in the small intestine were observed. With high II, all layers of the intestinal mucosa were affected and marked diffuse, subacute lymphoplasmacytic enteritis was noted. In ill mink, changes in the composition of the blood were revealed. The objective of our work was to develop an integrated approach to the diagnosis of mink eimeriidosis, including the study of the species composition of parasitic protozoa, the assessment of PR and II, the determination of the clinical and biochemical blood parameters of healthy and eimeriids infected animals, and the establishment of pathomorphological changes typical for eimeriosis and isosporosis that occur in chronic and asymptomatic form. In total, from 2013 to 2019, 6118 minks (Mustela vison, M. lutreola Linnaeus,1761, Neovison vison Schreber,1777) were studied in six fur-bearing animal farms of the North-Western region of the Russian Federation using the coprological method. Of these, 294 minks were studied intravitally (clinical study of animals, morphological and biochemical blood tests) and postmortem (autopsia of minks after euthanasia, histological and immunohistochemical tests). At the same time, parasitic fauna was studied in six animal farms of the North-West region of the Russian Federation, 2687 of the examined minks were infected, the prevalence rates (PR) were 43.92 %. It was found that two species of eimeria parasitize in minks, Eimeria vison and E. furonis and two isospores, Isospora laidlawi and I. eversmanni. The latter species was discovered by us in the Kaliningrad region of the Russian Federation for the first time. A deep sequencing of the V4 region of the 18S rDNA gene and bioinformatics analysis were performed, which made it possible to determine OTUs (operational taxonomic units) and establish coccidia’s taxonomic affiliation. Thus we were able to confirm the results of light microscopy and determine the taxonomic affiliation of the isolated oocysts. As a result of the analysis, it was found that the sequence of the E. vison DNA fragment of 383 bp is most similar (99.48 %) to the sequence of another species (E. ictide) found in the GenBank. Data on high morphological and genetic similarities raise the question of the taxonomic affiliation of these two species and require additional detailed study. Most often, eimeriidoses of minks proceeded in the form of mono infections (37.20 %), mixed infections with two parasites were 6.15 %, mixed infections with three protozoans made 0.57 % of cases. The peak of PR in young and adult minks occurred in the summer in the Northwestern region of the Russian Federation. In animals aged 1.5-6 months, eimeriosis and isosporosis proceeded mainly in acute and subacute forms, in minks older than 6 months — in subacute, chronic and latent. The content of hemoglobin and red blood cells in the blood of ill mink with eimeriidosis was significantly lower than in healthy minks, while the number of leukocytes, on the contrary, increased. Eosinophilia, segmented neutrophilia were also observed in ill animals, the number of basophils increased by 2 times, the content of stab neutrophils increased by 1.6 times. Proteinemia was observed in ill animals, the total bilirubin and creatinine content increased by 33.83 and 31.90 %, respectively, and the amount of urea decreased by 21.19 %. A histological examination of material from various parts of the intestine from animals infected with eimeriids revealed that at a low intensity of invasion (II) (in adult minks), although the disease was not clinically manifested in this group of animals, nevertheless, pathological changes in the histological level have already been recorded in small areas and were noted mainly only in the epithelial plate of the intestinal mucosa. With high II, damage to all layers of the intestinal mucosa was observed. Pronounced diffuse, subacute lymphoplasmacytic enteritis was discovered. The pathological processes caused by the parasitism of eimeriid in minks are often similar to those for various infectious diseases, such as the carnivorous plague virus, Aleutian mink disease and coronavirus born disease. To exclude the possibility of diagnostic errors, the material was sent to the laboratory for immunohistochemical studies (IHC), as a result of which antigens of the carnivorous plague virus, coronavirus and Aleutian mink disease were not detected. Nucleic acids of viruses were not detected in all studied samples; the result of IHC was negative in all samples.
Keywords: mink, eimeria, isospora, protozoa, pathogenesis, pathomorphology, histology, immunohistochemistry.
REFERENCES
- Anikanova B.C. Koktsidii zhivotnykh zverokhozyaistv Karelii. Avtoreferat kandidatskoi dissertatsii [Coccidia of farm animals in Karelia. PhD Thesis]. Petrozavodsk, 1991 (in Russ.).
- Kuznetsov Yu.E. Sovremennaya nauka: aktual'nye problemy i puti ikh resheniya, 2015, 1(14): 48-50 (in Russ.).
- Kuznetsov Yu.E. Kishechnye parazitozy pushnykh zverei v khozyaistvakh Leningradskoi oblasti. Kandidatskaya dissertatsiya [Intestinal parasitoses of fur animals in farms of Leningrad region. PhD Thesis]. St. Petersburg, 2012 (in Russ.).
- Safiullin R.T. Rossiiskii parazitologicheskii zhurnal, 2008, 2: 84-99 (in Russ.).
- Kuznetsov Yu.E. Mezhdunarodnyi vestnik veterinarii, 2019, 4: 43-50 (in Russ.).
- Hasnain S.Z., Gallagher A.L., Grencis R.K., Thornton D.J. A new role for mucins in immunity: insights from gastrointestinal nematode infection. The International Journal of Biochemistry & Cell Biology, 2013, 45(2): 364-374 CrossRef
- Turner J.E., Stockinger B., Helmby H. IL-22 mediates goblet cell hyperplasia and worm expulsion in intestinal helminth infection. PLoS Pathogens, 2013, 9(10): e1003698 CrossRef
- Chen T.-L., Chen S., Wu H.-W., Lee T.C., Lu Y.-Z., Wu L-.L., Ni Y.-H., Sun C.-H., Yu W.-H., Buret A.G., Yu L.C.-H. Persistent gut barrier damage and commensal bacterial influx following eradication of Giardia infection in mice. Gut Pathogens, 2013, 5(1): 26 CrossRef
- Kuznetsov Y.E., Engashev S.V., Engasheva E.S., Nikonov I.N., Kuznetsova N.V. Microbial community studying of the dogs' gastrointestinal tract by the T-RFLP molecular genetic method and assessing the natural resistance of animals. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 2018, 9(5): 1652-1660.
- Broadhurst M.J., Leung J.M., Kashyap V., McCune J.M., Mahadevan U., McKerrow J.H., Loke P. IL-22+ CD4+ T cells are associated with therapeutic Trichuris trichiura infection in an ulcerative colitis patient. Science Translational Medicine, 2010, 2(60): 60ra88 CrossRef
- Tsubokawa D., Ishiwata K., Goso Y., Yokoyama T., Kanuka H., Ishihara K., Nakamura T., Tsuji N. Induction of Sda-sialomucin and sulfated H-sulfomucin in mouse small intestinal mucosa by infection with parasitic helminth. Experimental Parasitology, 2015, 153: 165-173 CrossRef
- Poloz S.V. Aktual'nye problemy patologii sel'skokhozyaistvennykh zhivotnykh, 2000, 2: 403-405 (in Russ.).
- Jensen H.C. Coccidiose hos mink. Dansk Pelsdyravl, 1989, 8(52): 493.
- Klopfer U., Neuman M. A note on coccidiosis in minks. Refuah Veterinath, 1970, 3(27): 122-124.
- Heimesaat M.M., Bereswill S., Fischer A., Fuchs D., Struck D., Niebergall J., Jahn H.-K., Dunay I.R., Moter A., Gescher D.M., Schumann R.R., Göbel U.B., Liesenfeld O. Gram-negative bacteria aggravate murine small intestinal Th1-type immunopathology following oral infection with Toxoplasma gondii. The Journal of Immunology, 2006, 177(12): 8785-8795 CrossRef
- GOST 25383-82 (ST SEV 2547-80). Zhivotnye sel'skokhozyaistvennye. Metody laboratornoi diagnostiki koktsidioza [GOST 25383-82 (ST SEV 2547-80). Farm animals. Methods of laboratory diagnosis of coccidiosis]. Moscow, 1982 (in Russ.).
- MUK 4.2.3145-13. Laboratornaya diagnostika gel'mintozov i protozoozov [MUK 4.2.3145-13. Laboratory diagnosis of helminthiasis and protozoa]. Moscow, 2013 (in Russ.).
- Belova L.M., Gavrilova N.A., Pudovkin D.N., Tokarev A.N., Kuznetsov Yu.E. Zhidkost' dlya diagnostiki ootsist koktsidii, tsist balantidii i zhiardii, yaits gel'mintov raznykh klassov, kleshchei, nasekomykh, ikh otdel'nykh stadii razvitiya. Pat. RU 2472154 C2 (RF) MPK G01N33/48 A61D99/00. FGBOU VO «SPBGAVM» (RF). № 2010153464/13. Zayavl. 27.12.10. Opubl. 01.10.13. Byul. № 1 [A liquid for the diagnosis of coccidia oocysts, cysts of balantidia and giardia, helminth eggs of different classes, ticks, insects, their individual stages of development. Patent RU 2472154 C2 (RF) MPK G01N33/48 A61D99/00. FGBOU VO «SPBGAVM» (RF). № 2010153464/13. Appl. 27.12.10. Publ. 01.10.13. Bull. № 1] (in Russ.).
- Carreno R.A., Schnitzler B.E., Jeffries A.C., Tenter A.M., Johnson A.M., Barta J.R. Phylogenetic analysis of coccidia based on 18s rDNA sequence comparison indicates that Isospora is most closely related to Toxoplasma and Neospora. Journal of Eukaryotic Microbiology, 1998, 45(2): 184-188 CrossRef
- Wilcox J.J.S. Something for everyone: a review of «The biology and identification of the Coccidia (Apicomplexa) of carnivores of the world». The American Midland Naturalist, 2019, 181(1): 143-145 CrossRef
- Matsubayashi M., Takami K., Abe N., Kimata I., Tani H., Sasai K., Baba E. Molecular characterization of crane coccidia, Eimeria gruis and E. reichenowi, found in feces of migratory cranes. Parasitology Research, 2005, 97(1): 80-83 CrossRef
- Ogedengbe J.D., Hanner R.H., Barta J.R. DNA barcoding identifies Eimeria species and contributes to the phylogenetics of coccidian parasites (Eimeriorina, Apicomplexa, Alveolata). International Journal for Parasitology, 2011, 41(8): 843-850 CrossRef
- Pastor A.R. Investigating enteric coccidiosis in the black-footed (Mustela nigripes) and domestic ferret (Mustela putorius furo). PhD. University of Guelph, 2017.
- Immunogistokhimicheskie metody /Pod redaktsiei [Immunohistochemical methods. G.A. Frank, P.G. Mal'kov (eds.)]. Moscow, 2011 (in Russ.).
- Kuznetsov Yu.E., Belova L.M., Gavrilova N.A., Kuznetsova N.V., Shiryaeva V.A., Loginova O.A., Roberman M.G. Materialy III Mezhdunarodnogo parazitologicheskogo simpoziuma «Sovremennye problemy obshchei i chastnoi parazitologii» [Mat. III Int. Parasitological Symp. «Modern problems of general and private parasitology»]. St. Petersburg,2019: 164-168 (in Russ.).
- Gerasimchik V.A. Kishechnye parazitozy pushnykh zverei (etiologiya, epizootologiya, patogenez, diagnostika, terapiya i profilaktika). Avtoreferat kandidatskoi dissertatsii [Intestinal parasitoses of fur animals (etiology, epizootology, pathogenesis, diagnosis, therapy and prevention). PhD Thesis]. Minsk, 2008 (in Russ.).
- Duszynski D.W. Kvičerová J., Seville R.S. The biology and identification of the Coccidia (Apicomplexa) of Carnivores of the world. Academic Press,2018.
- Petersen H.H., Yang R., Chriél M., Hansen M.S., Ryan U.M. Morphological and molecular characterisation of Eimeria vison-like oocysts (Apicomplexa:Eimeriidae) in farmed mink (Neovison vison) in Denmark. Parasitol. Res.,2018, 117(9): 2933-2939 CrossRef