Diagnosis of Toxoplasmosis in Ruminants Aborted Fetuses in Northern Iran Using Molecular and Bioassay Techniques
-
Kaveh Azimi
,
Afsaneh Amouei
,
Mehdi Sharif
,
Shahabeddin Sarvi
,
Nemat Shams
,
Azadeh Mizani
,
Seyed Abdollah Hosseini
,
Saeid Salehi
,
Mohhamad Reza Safari-Kharyeki
,
Tooran Nayeri Chegeni
,
Zahra Hosseininejad
,
Ahmad Daryani
Abstract
Background: Toxoplasma gondii is a zoonotic obligatory intracellular protozoan parasite that infects a wide range of warm-blooded species. This study aimed to obtain further information on the role of T. gondii infection in ruminant abortion (sheep, goats and cattle) using bioassay and PCR methods in Mazandaran province, northern Iran.
Methods: Overall, 104 aborted fetuses (52 bovine, 48 ovine, 4 caprine) were collected at different stages of gestation during the lambing seasons in various parts of Mazandaran Province from Mar 2016 to May 2017. Brains of 104 aborted fetuses were bioassayed in female BALB/c mice. DNA was extracted from all brain samples using phenol-chloroform-isoamyl Alcohol instructions. RE gene was used for detection all of T. gondii DNA by conventional PCR assay.
Results: The results of the bioassayed samples were negative because no tachyzoites or cyst were observed in the peritoneal and brain specimens of the mice. The detection of T. gondii DNA was confirmed by observation of a 529 bp band in 15 out of 104 fetuses (14.4%). The highest prevalence rate of T. gondii detected from sheep (16.6%) followed by cattle (13.4%) and goats (0%). The highest prevalence of the infection was observed in east area, while the lowest prevalence of the infection was observed in west area.
Conclusion: T. gondii infection may partly be responsible for abortion and economic losses in livestock husbandry in this region. Therefore, further additional researches such as genotyping T. gondii and designing control strategies for improving management in livestock flocks are necessary.
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2. Heidari H, Gharekhani J, Tavoosidana G. Role of toxoplasmosis in abortion of ewes in western Iran: a serological study. Sci Parasitol. 2013; 14(2):99-103.
3. Dubey JP, Schares G. Neosporosis in animals—the last five years. Vet Parasitol. 2011; 180(1-2):90-108.
4. Tenter AM. Toxoplasma gondii in animals used for human consumption. Mem Inst Oswaldo Cruz. 2009; 104(2):364-9.
5. Dubey JP. Toxoplasmosis in sheep—the last 20 years. Vet Parasitol. 2009; 163(1-2):1-14.
6. Liu Q, Wang Z-D, Huang S-Y, et al. Diagnosis of toxoplasmosis and typing of Toxoplasma gondii. Parasit Vectors. 2015; 8:292.
7. Homan W, Vercammen M, De Braekeleer J, et al. Identification of a 200-to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR1. Int J Parasitol. 2000; 30(1):69-75.
8. De Sousa S, Ajzenberg D, Canada N, et al. Biologic and molecular characterization of Toxoplasma gondii isolates from pigs from Portugal. Vet Parasitol. 2006;135(2):133-6.
9. Dubey JP. Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol. 1998; 74(1):75-7.
10. Biase FH, Franco MM, Goulart LR, et al. Protocol for extraction of genomic DNA from swine solid tissues. Genet Mol Biol. 2002; 25(3):313-5.
11. Keshavarzi H, Sadeghi-Sefidmazgi A, Kristensen AR, et al. Abortion studies in Iranian dairy herds: I. Risk factors for abortion. Livest Sci. 2017; 195:45-52.
12. Munday B, Mason R. Toxoplasmosis as a cause of perinatal death in goats. Aust Vet J. 1979; 55(10):485-7.
13. Hartley W, Marshall S. Toxoplasmosis as a cause of ovine perinatal mortality. N Z Vet J. 1957; 5(4):119-24.
14. Garcia JL, Gennari SM, Machado RZ, et al. Toxoplasma gondii: detection by mouse bioassay, histopathology, and polymerase chain reaction in tissues from experimentally infected pigs. Exp Parasitol. 2006; 113(4):267-71.
15. da Silva AV, Langoni H. The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR). Vet Parasitol. 2001; 97(3):191-8.
16. Wastling J, Nicoll S, Buxton D. Comparison of two gene amplification methods for the detection of Toxoplasma gondii in experimentally infected sheep. J Med Microbiol. 1993; 38(5):360-5.
17. Amouei A, Sharif M, Sarvi S, et al. Aetiolo-gy of livestock fetal mortality in Mazanda-ran province, Iran. PeerJ. 2019;6:e5920.
18. Rassouli M, Razmi G, Bassami M, et al. Study on ovine abortion associated with Toxoplasma gondii in affected herds of Khorasan Razavi Province, Iran based on PCR detection of fetal brains and maternal serology. Parasitology. 2011; 138(6):691-7.
19. Habibi G, Imani A, Gholami M, et al. Detection and identification of Toxoplasma gondii type one infection in sheep aborted fetuses in Qazvin Province of Iran. Iran J Parasitol. 2012; 7(3):64-72.
20. Shahbazi G, RAD NH, Madani R, et al. Toxoplasma gondii in Aborted Fetuses of Sheep in Ardebil Area, North-West of Iran. Iran J Parasitol. 2019; 14(3): 430-435.
21. Kamani J, Egwu G, Mani A, et al. Survey of Toxoplasma gondii DNA in aborted Ovine and Caprine fetuses by nested PCR in Borno state, Nigeria. Vet World. 2010; 3(8):360-3.
22. Plant J, Beh K, Acland HM. laboratory findings form ovine abortion and perinatal mortality. Aust Vet J. 1972; 48(10):558-61.
23. Dubey JP, Kirkbride C. Toxoplasmosis and other causes of abortions in sheep from north central United States. J Am Vet Med Assoc. 1990; 196(2):287-90.
24. Pereira-Bueno J, Quintanilla-Gozalo A, Pérez-Pérez V, et al. Evaluation of ovine abortion associated with Toxoplasma gondii in Spain by different diagnostic techniques. Vet Parasitol. 2004; 121(1-2):33-43.
25. Charleston W. Toxoplasma and other protozoan infections of economic importance in New Zealand. N Z J Zool. 1994; 21:67-81.
26. Lélu M, Villena I, Dardé M-L, et al. Quantitative estimation of the viability of Toxoplasma gondii oocysts in soil. Appl Environ Microbiol. 2012; 78(15):5127-32.
27. Alvarado-Esquivel C, Campillo-Ruiz F, Liesenfeld O. Seroepidemiology of infection with Toxoplasma gondii in migrant agricultural workers living in poverty in Durango, Mexico. Parasit Vectors. 2013; 6:113.
28. Hamidinejat H, Goraninejad S, Ghorbanpoor M, et al. Role of Toxoplasma gondii in abortion of ewes in Ahvaz (South-West Iran). Bull Vet Inst Pulawy. 2008; 52(3):369-71.
29. Andrade MMC, Carneiro M, Medeiros AD, et al. Seroprevalence and risk factors associated with ovine toxoplasmosis in Northeast Brazil. Parasite. 2013; 20:20.
30. Fallahi S, Kazemi B, Bandehpour M, et al. Comparison of the RE and B1 gene for detection of Toxoplasma gondii infection in children with cancer. Parasitol Int. 2014; 63(1): 37-41.
31. Masala G, Porcu R, Daga C, et al. Detection of pathogens in ovine and caprine abortion samples from Sardinia, Italy, by PCR. J Vet Diagn Invest. 2007; 19(1):96-8.
32. Buxton D. Protozoan infections (Toxoplasma gondii, Neospora caninum and Sarcocystis spp.) in sheep and goats: recent advances. Vet Res. 1998; 29(3-4):289-310.
33. Innes EA, Bartley PM, Buxton D, et al. Ovine toxoplasmosis. Parasitology. 2009; 136(14):1887-94.
34. Scott P, Sargison N, Wilson D. The potential for improving welfare standards and productivity in United Kingdom sheep flocks using veterinary flock health plans. Vet J. 2007; 173(3):522-31.
35. Danehchin L, Razmi G, Naghibi A. Molecular detection of Toxoplasma gondii infection in aborted fetuses in sheep in Khorasan Razavi province, Iran. Iran J Vet Med. 2017; 11(2):147-53.
2. Heidari H, Gharekhani J, Tavoosidana G. Role of toxoplasmosis in abortion of ewes in western Iran: a serological study. Sci Parasitol. 2013; 14(2):99-103.
3. Dubey JP, Schares G. Neosporosis in animals—the last five years. Vet Parasitol. 2011; 180(1-2):90-108.
4. Tenter AM. Toxoplasma gondii in animals used for human consumption. Mem Inst Oswaldo Cruz. 2009; 104(2):364-9.
5. Dubey JP. Toxoplasmosis in sheep—the last 20 years. Vet Parasitol. 2009; 163(1-2):1-14.
6. Liu Q, Wang Z-D, Huang S-Y, et al. Diagnosis of toxoplasmosis and typing of Toxoplasma gondii. Parasit Vectors. 2015; 8:292.
7. Homan W, Vercammen M, De Braekeleer J, et al. Identification of a 200-to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR1. Int J Parasitol. 2000; 30(1):69-75.
8. De Sousa S, Ajzenberg D, Canada N, et al. Biologic and molecular characterization of Toxoplasma gondii isolates from pigs from Portugal. Vet Parasitol. 2006;135(2):133-6.
9. Dubey JP. Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol. 1998; 74(1):75-7.
10. Biase FH, Franco MM, Goulart LR, et al. Protocol for extraction of genomic DNA from swine solid tissues. Genet Mol Biol. 2002; 25(3):313-5.
11. Keshavarzi H, Sadeghi-Sefidmazgi A, Kristensen AR, et al. Abortion studies in Iranian dairy herds: I. Risk factors for abortion. Livest Sci. 2017; 195:45-52.
12. Munday B, Mason R. Toxoplasmosis as a cause of perinatal death in goats. Aust Vet J. 1979; 55(10):485-7.
13. Hartley W, Marshall S. Toxoplasmosis as a cause of ovine perinatal mortality. N Z Vet J. 1957; 5(4):119-24.
14. Garcia JL, Gennari SM, Machado RZ, et al. Toxoplasma gondii: detection by mouse bioassay, histopathology, and polymerase chain reaction in tissues from experimentally infected pigs. Exp Parasitol. 2006; 113(4):267-71.
15. da Silva AV, Langoni H. The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR). Vet Parasitol. 2001; 97(3):191-8.
16. Wastling J, Nicoll S, Buxton D. Comparison of two gene amplification methods for the detection of Toxoplasma gondii in experimentally infected sheep. J Med Microbiol. 1993; 38(5):360-5.
17. Amouei A, Sharif M, Sarvi S, et al. Aetiolo-gy of livestock fetal mortality in Mazanda-ran province, Iran. PeerJ. 2019;6:e5920.
18. Rassouli M, Razmi G, Bassami M, et al. Study on ovine abortion associated with Toxoplasma gondii in affected herds of Khorasan Razavi Province, Iran based on PCR detection of fetal brains and maternal serology. Parasitology. 2011; 138(6):691-7.
19. Habibi G, Imani A, Gholami M, et al. Detection and identification of Toxoplasma gondii type one infection in sheep aborted fetuses in Qazvin Province of Iran. Iran J Parasitol. 2012; 7(3):64-72.
20. Shahbazi G, RAD NH, Madani R, et al. Toxoplasma gondii in Aborted Fetuses of Sheep in Ardebil Area, North-West of Iran. Iran J Parasitol. 2019; 14(3): 430-435.
21. Kamani J, Egwu G, Mani A, et al. Survey of Toxoplasma gondii DNA in aborted Ovine and Caprine fetuses by nested PCR in Borno state, Nigeria. Vet World. 2010; 3(8):360-3.
22. Plant J, Beh K, Acland HM. laboratory findings form ovine abortion and perinatal mortality. Aust Vet J. 1972; 48(10):558-61.
23. Dubey JP, Kirkbride C. Toxoplasmosis and other causes of abortions in sheep from north central United States. J Am Vet Med Assoc. 1990; 196(2):287-90.
24. Pereira-Bueno J, Quintanilla-Gozalo A, Pérez-Pérez V, et al. Evaluation of ovine abortion associated with Toxoplasma gondii in Spain by different diagnostic techniques. Vet Parasitol. 2004; 121(1-2):33-43.
25. Charleston W. Toxoplasma and other protozoan infections of economic importance in New Zealand. N Z J Zool. 1994; 21:67-81.
26. Lélu M, Villena I, Dardé M-L, et al. Quantitative estimation of the viability of Toxoplasma gondii oocysts in soil. Appl Environ Microbiol. 2012; 78(15):5127-32.
27. Alvarado-Esquivel C, Campillo-Ruiz F, Liesenfeld O. Seroepidemiology of infection with Toxoplasma gondii in migrant agricultural workers living in poverty in Durango, Mexico. Parasit Vectors. 2013; 6:113.
28. Hamidinejat H, Goraninejad S, Ghorbanpoor M, et al. Role of Toxoplasma gondii in abortion of ewes in Ahvaz (South-West Iran). Bull Vet Inst Pulawy. 2008; 52(3):369-71.
29. Andrade MMC, Carneiro M, Medeiros AD, et al. Seroprevalence and risk factors associated with ovine toxoplasmosis in Northeast Brazil. Parasite. 2013; 20:20.
30. Fallahi S, Kazemi B, Bandehpour M, et al. Comparison of the RE and B1 gene for detection of Toxoplasma gondii infection in children with cancer. Parasitol Int. 2014; 63(1): 37-41.
31. Masala G, Porcu R, Daga C, et al. Detection of pathogens in ovine and caprine abortion samples from Sardinia, Italy, by PCR. J Vet Diagn Invest. 2007; 19(1):96-8.
32. Buxton D. Protozoan infections (Toxoplasma gondii, Neospora caninum and Sarcocystis spp.) in sheep and goats: recent advances. Vet Res. 1998; 29(3-4):289-310.
33. Innes EA, Bartley PM, Buxton D, et al. Ovine toxoplasmosis. Parasitology. 2009; 136(14):1887-94.
34. Scott P, Sargison N, Wilson D. The potential for improving welfare standards and productivity in United Kingdom sheep flocks using veterinary flock health plans. Vet J. 2007; 173(3):522-31.
35. Danehchin L, Razmi G, Naghibi A. Molecular detection of Toxoplasma gondii infection in aborted fetuses in sheep in Khorasan Razavi province, Iran. Iran J Vet Med. 2017; 11(2):147-53.
| Files | ||
| Issue | Vol 16 No 2 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v16i2.6273 | |
| Keywords | ||
| Toxoplasma gondii Aborted fetuses Sheep Goats Cattle Iran | ||
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How to Cite
1.
Azimi K, Amouei A, Sharif M, Sarvi S, Shams N, Mizani A, Hosseini SA, Salehi S, Safari-Kharyeki MR, Nayeri Chegeni T, Hosseininejad Z, Daryani A. Diagnosis of Toxoplasmosis in Ruminants Aborted Fetuses in Northern Iran Using Molecular and Bioassay Techniques. Iran J Parasitol. 2021;16(2):229-235.
