Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits

Strongyloidiasis in rabbits is a disease caused by Strongyloides papillosus helminths, manifested by skin inflammation, diarrhea, weight loss, and stunting of animals in growth and development. Confirmation of invasion is carried out mainly by required methods of helminthoovoscopy and helmintholarvo...

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Main Author: Y. V. Duda
Format: Article
Language:English
Published: Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv 2022-05-01
Series:Науковий вісник Львівського національного університету ветеринарної медицини та біотехнологій імені С.З. Ґжицького: Серія Ветеринарні науки
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Online Access:https://nvlvet.com.ua/index.php/journal/article/view/4322
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author Y. V. Duda
author_facet Y. V. Duda
author_sort Y. V. Duda
collection DOAJ
description Strongyloidiasis in rabbits is a disease caused by Strongyloides papillosus helminths, manifested by skin inflammation, diarrhea, weight loss, and stunting of animals in growth and development. Confirmation of invasion is carried out mainly by required methods of helminthoovoscopy and helmintholarvoscopy. The most well-known methods of helmintholarvoscopic diagnosis of feces for strongyloidiasis are the method of D. Baermann (1917) and its modifications according to V. I. Shilnikov (1983), the method using the “asterisk” device (according to V. F. Nikitin and T. Pavlasek, 1988), the method of coprohelmintho-larvoscopic rings (according to S. I. Ponomar, N. M. Soroka, 2007), the method of T. I. Popova, standardized by S. I. Ponomar (2003). There are no laboratory methods with 100 % reliability at the present stage. Therefore, our studies on improving the lifelong helminthocoprological diagnosis of strongyloidiasis in rabbits are relevant. Our work aimed to compare different methods of staining the pathogen S. papillosus. Laboratory studies were carried out in the laboratories of the Department of Parasitology and Veterinary Sanitary Expertise of the Dnipro State Agrarian and Economic University. For staining S. papillosus, 20 dyes were used: 1 % alizarin red, 1 % brilliant blue, Turk's solution, 1 % brilliant green, 1 % methylene blue, 1 % amido black 10B, 1 % eosin, Ziehl's solution, Lugol's solution, 1 % сarbolic gentian violet, Romanowsky-type, 1 % Sudan, 1 % bromophenol blue, 1 % orange G, 1 % bromocresol green, 1 % safranin, Zadorozhny-Dozmorov's-type, Mikhin's-type, Muromtsev's-type. We have proposed 20 dyes for staining the internal organs of larvae and free-living males and females pathogens of Strongyloides without or with temperature fixation. Live pathogens S. papillosus, without temperature fixation, pass only Lugol's and safranin's solutions. All other dyes showed the highest staining, but only when the preparation was heated for 2–3 seconds to 60 °C. However, the dyes showed different degrees of coloration. Bromocresol green stained the walls of the internal organs of the helminth. Namely, the nematodes acquired a green color, and the membranes of a part of the esophagus and intestines became dark green. During microscopy, this allowed accurate identification and differentiation into larvae and free-living males and females of S. papillosus and simplified their counting.
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series Науковий вісник Львівського національного університету ветеринарної медицини та біотехнологій імені С.З. Ґжицького: Серія Ветеринарні науки
spelling doaj.art-ae8b1a03bf0c42d59420aaeef9aa1b2b2022-12-22T02:35:20ZengStepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies LvivНауковий вісник Львівського національного університету ветеринарної медицини та біотехнологій імені С.З. Ґжицького: Серія Ветеринарні науки2518-75542518-13272022-05-01241059410110.32718/nvlvet105144322Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbitsY. V. Duda0Dnipro State Agrarian and Economic University, Dnipro, UkraineStrongyloidiasis in rabbits is a disease caused by Strongyloides papillosus helminths, manifested by skin inflammation, diarrhea, weight loss, and stunting of animals in growth and development. Confirmation of invasion is carried out mainly by required methods of helminthoovoscopy and helmintholarvoscopy. The most well-known methods of helmintholarvoscopic diagnosis of feces for strongyloidiasis are the method of D. Baermann (1917) and its modifications according to V. I. Shilnikov (1983), the method using the “asterisk” device (according to V. F. Nikitin and T. Pavlasek, 1988), the method of coprohelmintho-larvoscopic rings (according to S. I. Ponomar, N. M. Soroka, 2007), the method of T. I. Popova, standardized by S. I. Ponomar (2003). There are no laboratory methods with 100 % reliability at the present stage. Therefore, our studies on improving the lifelong helminthocoprological diagnosis of strongyloidiasis in rabbits are relevant. Our work aimed to compare different methods of staining the pathogen S. papillosus. Laboratory studies were carried out in the laboratories of the Department of Parasitology and Veterinary Sanitary Expertise of the Dnipro State Agrarian and Economic University. For staining S. papillosus, 20 dyes were used: 1 % alizarin red, 1 % brilliant blue, Turk's solution, 1 % brilliant green, 1 % methylene blue, 1 % amido black 10B, 1 % eosin, Ziehl's solution, Lugol's solution, 1 % сarbolic gentian violet, Romanowsky-type, 1 % Sudan, 1 % bromophenol blue, 1 % orange G, 1 % bromocresol green, 1 % safranin, Zadorozhny-Dozmorov's-type, Mikhin's-type, Muromtsev's-type. We have proposed 20 dyes for staining the internal organs of larvae and free-living males and females pathogens of Strongyloides without or with temperature fixation. Live pathogens S. papillosus, without temperature fixation, pass only Lugol's and safranin's solutions. All other dyes showed the highest staining, but only when the preparation was heated for 2–3 seconds to 60 °C. However, the dyes showed different degrees of coloration. Bromocresol green stained the walls of the internal organs of the helminth. Namely, the nematodes acquired a green color, and the membranes of a part of the esophagus and intestines became dark green. During microscopy, this allowed accurate identification and differentiation into larvae and free-living males and females of S. papillosus and simplified their counting.https://nvlvet.com.ua/index.php/journal/article/view/4322larva, free-living male and female, strongyloides papillosus, dye, rabbits
spellingShingle Y. V. Duda
Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
Науковий вісник Львівського національного університету ветеринарної медицини та біотехнологій імені С.З. Ґжицького: Серія Ветеринарні науки
larva, free-living male and female, strongyloides papillosus, dye, rabbits
title Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
title_full Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
title_fullStr Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
title_full_unstemmed Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
title_short Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits
title_sort comparison of different staining methods for the nematode strongyloides papillosus isolated from rabbits
topic larva, free-living male and female, strongyloides papillosus, dye, rabbits
url https://nvlvet.com.ua/index.php/journal/article/view/4322
work_keys_str_mv AT yvduda comparisonofdifferentstainingmethodsforthenematodestrongyloidespapillosusisolatedfromrabbits