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Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv
original article | UDC 636.92:616.99:574.2:614.91 doi: 10.32718/ujvas5-1.09
Volume 5 Number 1
Ovocidal action of disinfectants against eggs of Passalurus ambiguus
A. Khorolskyi
Poltava State Agrarian University, Skovorody Str., 1/3, Poltava, 36003, Ukraine
Article info Received 18.02.2022 Received in revised form
21.03.2022 Accepted 22.03.2022
Correspondence author Anatolii Khorolskyi Tel.: +38-095-568-72-56 E-mail: horoltolik1@gmail. com
2022 Khorolskyi A. This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Contents
1. Introduction................. .. 53
2. Materials and methods .... .. 54
3. Results and discussion .... .. 54
3.1. Results.................... . 54
3.2. Discussion................ .. 56
4. Conclusions................ .. 56
References ..................... 56
Abstract
Successful elimination of helminthic diseases, as well as their further prevention, is possible only under the condition of carrying out a complex of organizational and economic, veterinary and sanitary, and special anti-parasitic measures, which must necessarily include disinfestation of places where animals are kept, as well as environmental objects. It is necessary to establish the ability of modern chemical agents to harm certain types of parasites. The research aimed to determine the ovocidal efficiency of disinfectants in different concentrations and exposures against Passalurus ambiguus eggs isolated from domestic rabbits. An experimental test was carried out in laboratory conditions on chemical products of domestic production "Virosan". (AS - alkyldimethylbenzylammonium chloride, glutaraldehyde) and "Hermecid-VS" (AS - didecyldimethylammonium chloride, glutaraldehyde, ben-zalkonium chloride). The leading indicator of the effect of disinfectants on the eggs of pasaluris was the value of their disinfestation efficiency. The results of the conducted studies established a high level of ovocidal efficiency of "Virosan" (OE - 100 %) against P. ambiguus eggs at 0.25 % concentration for exposures of 30-60 minutes and 0.5 % concentration for exposures of 10-60 minutes. A satisfactory level of the ovocidal efficiency of "Virosan" was established when using the agent in 0.1 % (30-60 min exposure) and 0.25 % (10 min exposure) concentrations, OE ranges from 62.36 to 82.20 %. An unsatisfactory level of the ovocidal efficiency of "Virosan" was established at 0.1 % of the concentration after exposure for 10 minutes (AS - 47.30 %). Disinfectant "Hermecid-VS" showed a high level of ovocidal efficiency concerning the eggs of pasaluris in 0.1 % concentration for 60 min exposure (OE - 91.45 %) and in 0.25-0.5 % concentration for 10-60 min exposure (AS - 100 %). A satisfactory ovocidal efficiency of "Hermecid-VS" was established when using the product at a concentration of 0.1 % (10 and 30-minute exposures), where OE indicators were 76.25 and 81.91 %, respectively. The obtained data make it possible to recommend disinfectants of domestic production "Virosan" and "Hermecid-VS" in specified concentrations and exposures to control and prevent pasalurosis in rabbits in order to effectively carry out the deinvasion of environmental objects and places where animals are kept.
Keywords: rabbits, passalurosis, Passalurus ambiguus, nematode eggs, disinfectants, ovocidal action._
/ \ Citation:
Khorolskyi, A. (2022). Ovocidal action of disinfectants against eggs of Passalurus ambiguus. Ukrainian Journal of Veterinary and Agricultural Sciences, 5(1), 53-57.
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1. Introduction
Literary sources report that one of the essential factors in the transmission of animal helminthiasis pathogens is environmental objects where the exogenous development of embryonic and post-embryonic stages of parasites takes place (Dubna et al., 2007; Traversa et al., 2014; Wongrak et al., 2014; Moskvina et al., 2016). Therefore, preventive measures for helminthiasis should be aimed at neutralizing eggs and larvae at various stages of their development. The most well-known and widespread method is the deinvasion of environmental objects using various disinfectants (Tamasi, 1995; Mielke & Hiepe, 1998; Zoltowska et al., 2000).
Scientists from different countries of the world are constantly conducting experimental and production studies to determine the deinvasion properties of known disinfectants for various animal helminthiasis. Based on the obtained data, they determine the most effective chemical agents and recommend them for effective control and prevention of
parasitosis (Stromberg, 1997; Nowakowicz-D^bek et al., 2001; Tiersch et al., 2014).
In particular, the researchers found that the chemicals ethanol, methanol, and chlorhexidine did not affect the development of larvae of A. suum eggs, regardless of their concentration and treatment time. At the same time, 3 % cresol, 0.2 % sodium hypochlorite were retained but did not inactivate the eggs of pig roundworms. According to the authors, the investigated means did not destroy the albumen layer of A. suum eggs, which kept them viable (Oh et al., 2016).
Other scientists studied the effect of six disinfectants commonly used in nurseries and veterinary clinics on the embryogenesis and viability of T. canis eggs. They found that none of the tested agents had a significant inhibitory effect on the embryogenesis and viability of T. canis eggs, regardless of the time of contact (Ursache et al., 2019). Similar studies on T. canis eggs were conducted by the authors, who tested the ovocidal properties of benzalkonium chloride, 70 % ethanol, 2-2.5 % sodium hypochlorite solution,
and 7.99 % formaldehyde-based disinfectant. A high ovocidal action of 70 % ethanol was established, which led to 100 % death of Toxocara eggs. Sodium hypochlorite removed the outer layer of the eggs, but they contained motile larvae for up to 2 weeks. Benzalkonium chloride and formaldehyde-based disinfectants had no deleterious effects on T. canis embryogenesis (Verocai et al., 2010).
Therefore, for the effective implementation of preventive measures against animal helminthiasis, it is necessary to carry out deinvasion, which makes it impossible to infect the body with exogenous stages of parasite development (Labare et al., 2013; Zhang et al., 2020). However, the available literature lacks data on the disinfestation effect of modern disinfectants on the embryonic stages of development of nematodes of the species Passalurus ambiguus. Therefore, the aim of the research was to determine the ovocidal action of disinfectants in different concentrations and exposures against P. ambiguus eggs isolated from domestic rabbits.
2. Materials and methods
The work was carried out during the winter-spring period of 2022 based on the laboratory of the Department of Parasitology and Veterinary-Sanitary Examination of the Poltava State Agricultural University.
In order to determine the disinfestation efficiency of the disinfectants "Virosan" ("BioTestLab" Ltd, Ukraine) and "Hermecidu-VS" ("Vetsintez" Ltd, Ukraine), a test culture of nematode eggs of Passalurus ambiguus isolated from the gonads of female helminths was used. Mature nematodes were detected when dissecting the intestines of rabbits that came from private farms in the Poltava region.
Experimental Petri dishes with different concentrations of "Virosan" and "Hermecid-VS" (0.1 %, 0.25 %, and 0.5 %) were prepared, which were studied under different exposures (10, 30, 60 min). The same volume of a solution of a chemical agent of a particular concentration was added to the previously prepared mixture of eggs (at least 50 eggs). After the appropriate exposure, the culture of pasaluris eggs was washed four times in distilled water. Afterward, Petri dishes with washed egg cultures were placed in a thermostat at 35 °C and observed for five days. Egg culture was not treated with disinfectants and was used as a control. Each day, experimental and control egg cultures were examined under a microscope. Each experiment was repeated three times. The number of dead eggs per 50 found was counted. Indicators of ovocidal efficiency (OE, %) were determined. The assessment of disinfestation efficiency was carried out according to the indicators: high level of efficiency - 90100 %, satisfactory - 60-89 %, unsatisfactory - up to 60 %.
Mathematical analysis of the obtained data was carried out using the Microsoft "EXCEL" application program package by determining the arithmetic mean (M) and standard deviation (SD).
3. Results and discussion
3.1. Results
According to the results of the conducted research, a high level of ovocidal efficiency of "Virosan" (OE - 100 %) was established concerning the eggs of pasaluris at 0.25 %
concentration for exposures of 30-60 min and 0.5 % concentration for exposures of 10-60 min (Fig. 1).
0.1% 0.25% 0.5%
Fig. 1. Ovocidal efficiency (%) of "Virosan" against eggs of Passalurus ambiguus
At the same time, "Virosan" at a concentration of 0.1 % for exposures of 30-60 minutes and at a concentration of 0.25 % for exposures of 10 minutes showed a satisfactory level of ovocidal efficiency against the eggs of pasaluris (62.36 ± 5.93-68.98 ± 7.58 % and 82.20 ± 6.45 %, respectively). The remedy at a concentration of 0.1 % after exposure for 10 minutes was ineffective; OE did not exceed 47.30 ± 6.88 %.
In the process of embryogenesis of P. ambiguus under the influence of 0.1 % "Virosan" solution for 10 min exposure, the formation of larvae was established in 18.67 ± 3.06 eggs, in 30 min - in 13.33 ± 2.52 eggs, in 60 min - in 11.00 ± 3.00 pasaluris eggs. When using a 0.25 % solution of the agent, development was established only in 6.33 ± 2.52 eggs of pasaluris, in which larvae were formed (Table 1).
Later, with increasing exposure and concentration, 100% of the eggs stopped their development due to the destruction of the shell at the zygote stage (Fig. 2), as well as the resorption of the larva in the egg at the stage of its formation (Fig. 3).
Disinfectant "Hermecid-VS" showed a high level of ovocidal efficiency in relation to the eggs of pasaluris in 0.1 % concentration during exposure for 60 minutes (OE -91.45 ± 3.02 %) and in 0.25-0.5 % concentration during exposure 10-60 min (OE - 100 %). At the same time, "Hermecid-VS" in a concentration of 0.1 % for exposures of 10 and 30 minutes showed a satisfactory level of ovocidal efficiency against pasaluris eggs, where OE indicators were 76.25 ± 6.61 and 81.91 ± 6.40 %, respectively (Fig. 4).
In the process of embryogenesis of P. ambiguus under the action of 0.1 % solution of "Hermecid-VS" for 10 minutes exposure, further development with the formation of larvae was established in 8.33 ± 2.08 eggs. During exposures of 30 and 60 minutes, the number of eggs that continued to develop with the formation of larvae was at the level of 6.33 ± 2.08 and 3.00 ± 1.00 specimens, respectively (Table 2).
Later, with increasing exposure and concentration, 100 % of eggs stopped their development in the process of their embryogenesis. The deleterious effect of the agent was characterized by the accumulation of air bubbles under the shell of pasaluris eggs at the zygote stage and the stage of crushing and formation of blastomeres (Fig. 5).
Table 1
Indicators of the ovocidal activity of "Virosan" against Passalurus ambiguus eggs during their embryogenesis in laboratory conditions (M ± SD, min-max)
Exposition, Indicators Concentration of the solution
min 0.1 % 0.25 % 0.5 % Control
10.00 Larva formation 18.67 ± 3.06 /16-22/ 6.33 ± 2.52 /4-9/ - 35.33 ± 1.53 /34-37/
Death of eggs 31.33 ± 3.06 /28-34/ 43.67 ± 2.52/ /41-46/ 50 14.67 ± 1.53 /13-16/
30.00 Larva formation 13.33 ± 2.52 /11-16/ - - 35.33 ± 1.53 /34-37/
Death of eggs 36.67 ± 2.52 /34-39/ 50 50 14.67 ± 1.53 /13-16/
60.00 Larva formation 11.00 ± 3.00 /8-14/ - - 35.33 ± 1.53 /34-37/
Death of eggs 39.00 ± 3.00 /36-42/ 50 50 14.67 ± 1.53 /13-16/
ШЙ
1
%
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Jy ' 1 ! / •
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V
50 um
Fig. 2. Destruction of the shell in the egg of Passalurus ambiguus at the zygote stage with the use of "Virosan"
Ш
50 um
Fig. 3. Resorption of the larva in the egg of Passalurus ambiguus at the stage of its formation using "Virosan"
100 80 60 40 20' 0
60 min min
0.1%
0.25%
0.5%
Fig. 4. Ovocidal activity (%) of "Hermecid-VS" against
eggs of Passalurus ambiguus
Fig. 5. The appearance and accumulation of air bubbles under the shell of Passalurus ambiguus eggs at the zygote and crushing stages and the formation of blastomeres with the use of "Hermecid-VS"
Table 2
Indicators of the ovocidal activity of "Hermecid-VS" against Passalurus ambiguus eggs during their embryogenesis in laboratory conditions (M ± SD, min-max)
Exposition, Indicators Concentration of the solution
min 0.1 % 0.25 % 0.5 % Control
10.00 Larva formation 8.33 ± 2.08 /6-10/ - - 35.33 ± 1.53 /34-37/
Death of eggs 41.67 ± 2.08 /40-44/ 50 50 14.67 ± 1.53 /13-16/
30.00 Larva formation 6.33 ± 2.08 /4-8/ - - 35.33 ± 1.53 /34-37/
Death of eggs 43.67 ± 2.08 /42-46 50 50 14.67 ± 1.53 /13-16/
60.00 Larva formation 3.00 ± 1.00 /2-4/ - - 35.33 ± 1.53 /34-37/
Death of eggs 47.00 ± 1.00 /46-18/ 50 50 14.67 ± 1.53 /13-16/
Therefore, the disinfectants "Virosan" and "Hermecid-VS" have ovocidal activity against the eggs of pasaluris, which parasitize rabbits. At the same time, the indicators of their ovocidal efficiency depend on the concentration of chemicals and exposure to their use.
3.2. Discussion
Literary sources report the importance of deinvasion as an effective method of combating and preventing helminthi-asis in livestock and poultry farming. Moreover, most manufacturers offer disinfectants that do not have information about their ovocidal efficiency in their instructions (Tamasi, 1995; Zhu et al., 2013; Tiersch et al., 2014). Therefore, most scientists note the importance of determining the effectiveness of disinfectants in relation to exogenous stages of parasite development for certain helminthiasis (Moazeni et al., 2017; Zazharskyi et al., 2018; Ursache et al., 2019; Melny-chuk et al., 2020).
Our studies have established the ovocidal properties of modern disinfectants against the eggs of nematodes of the species Passalurus ambiguus, namely: "Virosan" (AS -alkyldimethylbenzylammonium chloride, glutaraldehyde) and "Hermecid-VS" (DR - didecyldimethylammonium chloride, glutaraldehyde, benzalkonium chloride). The high ovocidal efficiency of "Virosan" has been proven in 0.25 % (30-60 min exposure) and 0.5 % (10-60 min exposure) concentrations, where OE reaches 100 %. At the same time, a high level of ovocidal efficiency of "Hermecid-VS" concerning the eggs of pasaluris was established at 0.1 % concentration after exposure for 60 minutes (OE - 91.45 %) and at 0.25-0.5 % concentration after exposure for 10-60 min (OE - 100 %).
Scientists also proved the ovocidal properties of "Viro-san" in relation to eggs of Trichuris isolated from sheep. They established a high level of disinfestation efficiency of the agent (OE is 90.59-100.00 %) for the species Trichuris globulosa - in concentrations of 0.25 % (exposure for 60 minutes), 0.5-1.0 % (10-60 minutes); for the species T. skrjabini, T. ovis - in 0.5-1.0 % concentrations (10-60 min) (Melnychuk & Yuskiv, 2018).
The obtained data allow recommending disinfectants of domestic production "Virosan" and "Hermecid-VS" in specified concentrations and exposures for the control and prevention of pasalurosis in rabbits in order to effectively carry out the deinvasion of environmental objects and places where animals are kept.
4. Conclusions
A high level of the ovocidal efficiency of "Virosan" against the eggs of pasaluris isolated from rabbits was established when it was used in concentrations of 0.25 and 0.5 % for exposures of 30-60 and 10-60 min, respectively (100 %). The agent "Hermecid-VS" showed a high level of ovocidal efficiency at a concentration of 0.1 % for exposure of 60 minutes (91.45 %) and in concentrations of 0.25-0.5 % for exposures of 10, 30 and 60 minutes (100 %) relative to the embryonic stages of development of Passalurus ambiguus.
Conflict of interest.
The author state that there is no conflict of interest. References
Dubna, S., Langrova, I., Jankovska, I., Vadlejcha, J., Pekarb, S., Napravnika, J., & Fechtner, J. (2007). Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic. Veterinary Parasitology, 144(1-2), 81-86. [Crossref] [Google Scholar] Labare, M. P., Soohoo, H., Kim, D., Tsoi, K., Liotta, J. L., & Bowman, D. D. (2013). Ineffectiveness of a quaternary ammonium salt and povidone-iodine for the inactivation of Ascaris suum eggs. American Gournal of Infection Control, 41(4), 360-361. [Crossref] [Google Scholar] Melnychuk, V. V., Yuskiv, I. D., & Pishchalenko, M. A. (2020). Ovo-cidal action of glutaraldehyde and benzalkonium chloride mixture on Aonchotheca bovis (Nematoda, Capillariidae) embryogenesis. Regulatory Mechanisms in Biosystems, 11(2), 175-179. [Crossref] [Google Scholar] Melnychuk, V., & Yuskiv, I. (2018). Studying of disinvasion action of the disinfectant Virosan for eggs Nematodes genus Trichuris parasitizing in sheep. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 20(88), 16-23. [Crossref] [Google Scholar] Mielke, D., & Hiepe, T. (1998). The effectiveness of different disinfectants based on p-chloro-m-cresol against Ascaris suum eggs under laboratory conditions. Berliner und Münchener Tierärztliche Wochenschrift, 111(7-8), 291-294. [Abstract] [Google Scholar] Moazeni, M., Saadaty Ardakani, Z. S., Saharkhiz, M. J., Jalaei, J., Khademolhoseini, A. A., Shams Esfand Abad, S., & Mootabi Alavi, A. (2017). In vitro ovicidal activity of Peganum harma-la seeds extract on the eggs of Fasciola hepatica. Journal of Parasitic Diseases, 41(2), 467-472. [Crossref] [Google Scholar]
Moskvina, T. V., Bartkova, A. D., & Ermolenko, A. V. (2016). Geohelminths eggs contamination of sandpits in Vladivostok, Russia. Asian Pacific Journal of Tropical Medicine, 9(12), 1215-1217.
[Crossref] [Google Scholar! Nowakowicz-D^bek, B., Ondrasovic, M., Bis-Wencel, H., & Saba, L. (2001). Soil pollution with parasite eggs and larvae at fur-bearing animal farms. Medycyna Weterynaryjna, 57(3), 202-203. [Google Scholar]
Oh, K. S., Kim, G. T., Ahn, K. S., & Shin, S. S. (2016). Effects of disinfectants on larval development of Ascaris suum eggs. Korean journal of parasitology, 54(1), 103-107. [Crossref] [Google Scholar] Stromberg, B. E. (1997). Environmental factors influencing transmission. Veterinary Parasitology, 72(3-4), 247-256. [Crossref] [Google Scholar] Tamasi, G. (1995). Testing disinfectants for efficacy. Scientific and Technical Review, 14(1), 75-79. [Abstract] [Google Scholar] Tiersch, K. M., Da§, G., von Samson-Himmelstjerna, G., & Gauly, M. (2014). Artificial infection of chickens with Capillaria ob-signata eggs embryonated in different media. Veterinary Para-sitology, 200(1-2), 139-146. [Crossref] [Google Scholar] Traversa, D., Frangipane di Regalbono, A., Di Cesare, A., La Torre, F., Drake, J., & Pietrobelli, M. (2014). Environmental contamination by canine geohelminths. Parasites & Vectors, 7, 67. [Crossref] [Google Scholar] Ursache, A. L., Mircean, V., Dumitrache, M., Andrei, S., §tefam4, L., Cozma, V., Cätanä, R., & Cernea, M. (2019). Is routine disinfection efficient in preventing contamination with Toxocara canis eggs? Journal of Helminthology, 94, e60. [Crossref] [Google Scholar]
Verocai, G. G., Tavares, P. V., Ribeiro, F., Correia, T. R., & Scott, F. B. (2010). Effects of disinfectants on Toxocara canis embryogenesis and larval establishment in mice tissues. Zoonoses and Public Health, 57(7-8), 213-216. [Crossref] [Google Scholar] Wongrak, K., Da§, G., Moors, E., Sohnrey, B., & Gauly, M. (2014). Establishment of gastro-intestinal helminth infections in freerange chickens: a longitudinal on farm study. Berliner undMun-chener Tierarztliche Wochenschrift, 127(7-8), 314-321. [Abstract] [Google Scholar] Zazharskyi, V. V., Davydenko, P., Kulishenko, O., Chumak, V., Kryvaya, A., Biben, I. A., Tishkina, N. M., Borovik, I., Boyko, O. O., & Brygadyrenko, V. V. (2018). Bactericidal, protisto-cidal and nematodicidal properties of mixtures of alkyldime-thylbenzyl ammonium chloride, didecyldimethyl ammonium chloride, glutaraldehyde and formaldehyde. Regulatory Mechanisms in Biosystems, 9(4), 540-545. [Crossref] [Google Scholar] Zhang, S., Angel, C., Gu, X., Liu, Y., Li, Y., Wang, L., Zhou, X., He, R., Peng, X., Yang, G., & Xie, Y. (2020). Efficacy of a chlorocresol-based disinfectant product on Toxocara canis eggs. Parasitology Research, 119(10), 3369-3376. [Crossref] [Google Scholar] Zhu, L., Dai, J .L., Yang, L., & Qiu, J. (2013). In vitro ovicidal and larvicidal activity of the essential oil of Artemisia lancea against Haemonchus contortus (Strongylida). Veterinary Parasitology, 195(1-2), 112-117. [Crossref] [Google Scholar] Zôltowska, K., Bialowas, K., & Lopienska, E. (2000). Influence of zinc and lead ions on the development of eggs of Ascaris suum (Nematoda). Wiadomosci Parazytologiczne, 46(4), 501-506. [Abstract] [Google Scholar]
