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


Keywords: larva, free-living male and female, Strongyloides papillosus, dye, rabbits

Abstract

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.

Downloads

Download data is not yet available.

References

Areskog, M., Sollenberg, S., Engström, A., Samson-Himmelstjerna, G., & Höglund, J. (2014). A controlled study on gastrointestinal nematodes from two Swedish cattle farms showing field evidence of ivermectin resistance. Parasites and Vectors, 7, 13. DOI: 10.1186/1756-3305-7-13.

Boyko, O. O., Duda, Y. V., Pakhomov, O. Y., & Brygadyrenko, V. V. (2016). Comparative analysis of different methods of staining the larvae Haemonchus contortus, Mullerius sp. (Nematoda, Strongylida) and Strongyloides papillosus (Nematoda, Rhabditida). Folia Oecologica, 43(2), 129–137. URL: https://dspace.dsau.dp.ua/bitstream/123456789/119/1/Boyko%205.pdf.

Dakhno, I. S., & Lazorenko, L. M. (2010). Efektyvnist koproovoskopichnykh metodiv diahnostyky nema-todoziv u konei. Naukovyi visnyk Lvivskoho natsionalnoho universytetu veterynarnoi medytsyny ta biotekhnolohii imeni S. Z. Gzhytskoho. Seriia: Vet-erynarni nauky, 12(2(44), 71–73 (in Ukrainian).

Derkachev, D. Ju., Orobec, V. A., & Zaichenko, I. V. (2014). Sravnitel'naja ocenka jeffektivnosti kolich-estvennyh metodov koproovoskopii. Rossijskij parazitologicheskij zhurnal, 3, 68–73. URL: https://vniigis.elpub.ru/jour/article/view/258/261 (in Russian).

DeRosa, A. A., Chirgwin, S. R., Williams, J. C., & Klei, T. R. (2008). Isolation and characterization of a gene encoding carbonic anhydrase from Ostertagia ostertagi and quantitative measurement of expression during in vivo exsheathment. Veterinary Parasitology, 154(1-2), 58–66. DOI: 10.1016/j.vetpar.2008.02.021.

Dovhii, Yu., Vakhovskyi, I., Didkivskyi, O., Zhuravlova, O., & Zhuravlov, V. (2004). Patent Ukrainy № 58688. Kyiv: Ukrainskyi instytut intelektualnoi vlastnostі. URL: https://uapatents.com/?search=58688&type=number (in Ukrainian).

Duda, Y. V., & Kuneva, L. V. (2020). Diagnostika i se-zonnaja dinamika strongiloidoza u krolikov. Uchenye Zapiski uchrezhdenija obrazovanija Vitebskoj ordena “Znak pocheta” gosudarstvennoj akademii veterinar-noj mediciny, 56(1), 33–38. URL: https://dspace.dsau.dp.ua/ handle/123456789/3112 (in Russian).

Duda, Y. V., Kunieva, L. V., Shevchyk, R. S., & Taibov, E. E. (2018). Vplyv Strongyloides papillosus na mor-folohichni pokaznyky krovi kroliv. Vynakhidnytstvo ta ratsionalizatorstvo u medytsyni, biolohii ta ekolo-hii: Mizhnarodna naukovo-praktychna konferentsiia studentiv ta molodykh vchenykh, Dnipro, 19–20 veresnia 2018: tezy dopovidi, Dnipro, 48–49. URL: https://dspace.dsau.dp.ua/handle/123456789/2864 (in Ukrainian).

Duda, Y. V., Prus, M. P., & Lytvynenko, O. P. (2020). Naukovo-praktychni rekomendatsii z diahnostyky ta zakhodiv borotby z osnovnymy shlunkovo-kyshkovymy parazytozamy kroliv, Dnipro (in Ukrain-ian).

Huhosian, Y. A. (2018). Stronhiloidoz konei (poshyrennia, diahnostyka, zakhody borotby): avtoref. na zdobuttia nauk. stupenia kand. vet. nauk: 16.00.11. Lviv. nats. un-t vet. medytsyny ta bio-tekhnolohii im. S. Z. Gzhytskoho, Lviv URL: https://lvet.edu.ua/images/step/2018/04/25/ avtoref_huhosjyan.pdf (in Ukrainian).

Ievstafieva, V. O., Halat, V. F., & Halat, M. V. (2007). Zastosuvannia lichylnoi kamery dlia zazhyttievoi di-ahnostyky invaziinykh khvorob. Visnyk Derzhavno-ho ahroekolohichnoho universytetu, 2(19), 260–265. URL: http://ir.znau.edu.ua/bitstream/123456789/6578/1/VZNAU_2007_2_1_260-265.pdf (in Ukrainian).

Kobayashi, I., & Horii, Y. (2008). Gastrointestinal motor disturbance in rabbits experimentally infected with Strongyloides papillosus. Veterinary Parasitology, 158(1-2), 67–72. DOI: 10.1016/j.vetpar.2008.08.017.

Korchan, L. M., & Korchan, M. I. (2011). Porivnialna efektyvnist okremykh helmintolarvoskopichnykh sposobiv diahnostyky lehenevykh nematodoziv u dribnoi rohatoi khudoby. Visnyk Poltavskoi derzhavnoi ahrarnoi akademii. Poltava, 3, 117–119 URL: https://www.pdau.edu.ua/sites/default/files/ visnyk/2011/03/117.pdf (in Ukrainian).

Kotel'nikov, G. A. (1984). Gel'mintologicheskie issledo-vanija zhivotnyh i okruzhajushhej sredy. M.: Kolos (in Russian).

Larsson, A., Dimander, S. O., Waller, A., Uggla, A., & Höglund, J. (2007). A 3-year field evaluation of pasture rotation and supplementary feeding to control parasite infection in first-season grazing cattle – dynamics of pasture infectivity. Veterinary Parasitology, 145(1-2), 129–137. DOI: 10.1016/j.vetpar.2006.12.005.

Liashchenko Ye., Shendryk Kh., & Soroka N. (2012). Patent Ukrainy № 69062. Kyiv: Ukrainskyi instytut intelektualnoi vlastnostі. URL: https://uapatents.com/5-69062-sposib-pidrakhunku-yaehc-gelmintiv-u-fekaliyakh. html (in Ukrainian).

Nakamura, Y., & Motokawa, M. (2000). Hypolipemia associated with the wasting condition of rabbits infected with Strongyloides papillosus. Veterinary Parasitology, 88(1–2), 147–151. DOI: 10.1016/s0304-4017(99)00194-6.

Novitska, O. V., & Semenko, O. V. (2015). Zarazni khvoroby kroliv. K: TOV NVP “Interservis” (in Ukrainian).

Ponomar, S. I., Artemenko, L. P., Lytvynenko, O. P., & Honcharenko, V. P. (2011). Dovidnyk z labora-tornykh metodiv diahnostyky invaziinykh khvorob tvaryn. Bila Tserkva (in Ukrainian).

Ponomar, S. I., Honcharenko, V. P., & Soloviova, L. M. (2010). Dovidnyk z dyferentsiiuvannia zbudnykiv invaziinykh khvorob tvaryn. Kyiv: Ahrarna osvita (in Ukrainian).

Ponomar, S. I., Soroka, N. M., Lytvynenko, O. P., An-tipov, A. A., Honcharenko, V. P., Artemenko, L. P., Nebeshchuk, D., Soloviova, L. M., Palamarchuk, O. V., Nebeshchuk, L. V., & Yerokhina, O. M. (2008). Rekomendatsii shchodo helmintolohichnykh doslidzhen tvaryn. Bila Tserkva (in Ukrainian).

Ponomar, S. I., Soroka, N. M., Nebeshchuk, O. D., Hon-charenko, V. P., Semenko, O. V., & Ponomar, Z. S. (2015). Dovidnyk z vyznachennia helmintiv tvaryn. Bila Tserkva: TOV “Ofset” (in Ukrainian).

Prus, M. P., & Duda, Y. V. (2021). Zbudnyky khvorob travnoho kanalu kroliv u skladi parazytotsenoziv. Naukovyi visnyk Lvivskoho natsionalnoho univer-sytetu veterynarnoi medytsyny ta biotekhnolohii imeni S. Z. Gzhytskoho. Seriia: Veterynarni nauky, 23(102), 93–98. DOI: 10.32718/nvlvet10214 (in Ukrainian).

Prykhodko, Y. O., Byrka, V. I., Fedorova, O. V. Ponoma-renko, V. I., Mazannyi, O. V., Ponomarenko, A. M., & Nikiforova, O. V. (2017). Laboratorna diahnostyka invaziinykh khvorob tvaryn: metod. rekom. Kh. (in Ukrainian).

Safiullin, R. T., Shibitov, S. K., & Kotkov, A. V. (2008). Sistema probopodgotovki dlja parazitologicheskih is-sledovanij «Parasep» i ee aprobacija dlja diagnostiki gel'mintozov svinej. Rossijskij parazitologicheskij zhurnal, 3, 81–86. URL: https://cyberleninka.ru/article/n/sistema-probopodgotovki-dlya-parazitologicheskih-issledovaniy-parasep-i-ee-aprobatsiya-dlya-diagnostiki-gelmintozov-sviney (in Russian).

Siddiqui, A. A., & Berk, S. L. (2001). Diagnosis of Strongyloides stercoralis infection. Clinical Infectious Diseases, 33(7), 1040–1047. DOI: 10.1086/322707.

Srisuphanunt, M., Saksirisampant, W., & Karanis, Р. (2009). Detection of Cryptosporidium oocysts in green mussels (Perna viridis) from shell-fish markets of Thailand. Parasite, 16(3), 235–239. DOI: 10.1051/parasite/2009163235.

Taira, N., Minami, T., & Smitanon, J. (1991). Dynamics of faecal egg counts in rabbits experimentally infected with Strongyloides papillosus. Veterinary Parasitolo-gy, 39(3–4), 333–336. DOI: 10.1016/0304-4017(91)90050-6.

Thamsborg, S., Ketzis, J., Horii, Y., & Matthews, J. (2017). Strongyloides spp. infections of veterinary im-portance. Parasitology, 144(3), 274–284. DOI: 10.1017/S0031182016001116.

Torgerson, P. R., Paul, M., & Furrer, R. (2014). Evaluating faecal egg count reduction using a specifically de-signed package “eggCounts” in R and a user friendly web interface. Int. J. Parasitol, 44(5), 299–303. DOI: 10.1016/j.ijpara.2014.01.005.

Trach, V. N. (1992). Rekomendacii po primeneniju novo-go metoda ucheta jaic gel'mintov i cist prostejshih v fekalijah zhivotnyh. Kiev: Gosagroprom USSR (in Russian).

Yevstafieva, V. O. (2007). Patent Ukrainy № 26038. Kyiv: Ukrainskyi instytut intelektualnoi vlastnostі. URL: https://uapatents.com/?search=26038&type=number (in Ukrainian).

Zajac, A. M., & Conboy, G. A. (2012). Veterinary clinical parasitology 8th ed. UK John Wiley Sons Ltd.

Zeeshan, M., Zafar, A., & Saeed, Z. (2011). Use of “Para-sep filter fecal concentrator tubes” for the detection of intestinal parasites in stool samples under routine conditions. Indian Journal of Pathologyand Microbi-ology, 54(1), 121–123. DOI: 10.4103/0377-4929.77358

Abstract views: 0
PDF Downloads: 0
Published
2022-05-04
How to Cite
Duda, Y. (2022). Comparison of different staining methods for the nematode Strongyloides papillosus isolated from rabbits. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 24(105), 94-101. https://doi.org/10.32718/nvlvet10514