Molecular Detection of Neospora Canium in Bovine Colostrum: A Possible Risk Factor for Neonatal Infection
Document Type : Original Article
Authors
- Khadijeh Seydi-Gazafi 1
- Ali Sadeghi-nasab 2
- Hossein Rezvan 1
- Mehdi Namavari 3
- Sahar Hamoonnavard 1
- Bahram Dadmehr 4
1 Department of Pathobiology, Faculty of Veterinary Medicine, Bu-Ali Sina University, Hamedan, Iran
2 Department of Clinical Medicine, Faculty of Veterinary Medicine, Bu-Ali Sina University, Hamedan, Iran
3 Razi Vaccine and Serum Research Institute, Shiraz, Iran
4 Department of Basic Sciences, Applied Science Education Center of Farab Amarlou, Jirandeh, Iran
10.22084/avr.2025.30999.1004
Abstract
Neospora caninum is an obligate intracellular parasite that is both vertically and horizontally transmitted in cattle leading to signi cant economic losses in the cattle industry. The aim of this study was to investigate the prevalence of colostrum contamination in Neospora positive cows as a potential route for horizontal transmission. To this end, whole blood and colostrum from 44 fresh cows were analyzed using polymerase chain reaction (PCR) and nested PCR methods to detect the presence of the parasite DNA. In addition, serum samples were tested with a whole cell-based enzyme-linked immunosorbent assay (ELISA) to identify antibodies against the parasite. The results showed that 19 out of 44 colostrum samples (43.18%) contained the parasite genome. In addition, 16 of 18 (88.9%) Neospora -positive cows and 13 of 14 (92.9%) seropositive cows were found to have secreted infected colostrum. The relative risk (RR) of colostrum infection was signi cantly higher in blood PCR-positive cows (RR=7.7, P=0.0002) and seropositive cows (RR=4.6, P<0.0001) compared to uninfected cows. Based on these results, it is suggested that colostrum may be considered as a potential source of horizontal transmission in newborn calves and that methods for removing the infection such as pasteurisation may be required to reduce the risk of transmission.
Keywords
[1] Dubey JP, Schares G. Neosporosis in animalsthe
last ve years. Veterinary parasitology. 2011 Aug
4; 180(1-2): 90-108.
[2] Wouda W, Moen AR, Schukken YH. Abortion risk
in progeny of cows after a Neospora caninum epi-
demic. Theriogenology. 1998 May 1; 49(7): 1311-6.
[3] Stelwagen K, Carpenter E, Haigh B, Hodgkinson
A, Wheeler TT. Immune components of bovine
colostrum and milk. Journal of animal science.
2009 Apr 1; 87 (suppl 13): 3-9.
[4] Marin MS, Hecker YP, Quintana S, Perez SE,
Leunda MR, Canton GJ, Cobo ER, Moore DP,
Odeon AC. Toll-like receptors 3, 7 and 8 are upreg-
ulated in the placental caruncle and fetal spleen of
Neospora caninum experimentally infected cattle.
Veterinary Parasitology. 2017 Mar 15; 236: 58-61.
[5] Rivera JM, Hecker YP, Burucua MM, Cirone KM,
Cheuquepan FA, Fiorani F, Dorsch MA, Colque
LA, Canton GJ, Marin MS, Moore DP. Innate
and humoral immune parameters at delivery in
colostrum and calves from heifers experimentally
infected with Neospora caninum. Molecular Im-
munology. 2021 Apr 1; 132: 53-9.
[6] Moskwa B, Pastusiak K, Bien J, Cabaj W. The
rst detection of Neospora caninum DNA in the
colostrum of infected cows. Parasitology Research.
2007 Feb; 100: 633-6.
[7] Lindsay DS, Dubey JP. Infections in mice with
tachyzoites and bradyzoites of Neospora caninum
(Protozoa: Apicomplexa). The Journal of para-
sitology. 1990 Jun 1: 410-3.
[8] Moskwa B, Cabaj W. The role of the colostrum
and milk in Neospora caninum transmission.
Helminthologia. 2007 Sep; 44: 126-9.
[9] Mohammed NQ, Aaiz NN. Serological and molec-
ular investigation of Neospora caninum in cattle
in Al-Diwaniyah province, Iraq. Open Veterinary
Journal. 2025 Apr 30; 15(4): 1645.
[10] Ayati R, Rezvan H, Namavari MM, Sadeghi-nasab
A, Hamoon Navard S, Dadmehr B. Association be-
tween presence of DNA and antibody in the serum
during vertical transmission of Neospora caninum.
Iranian Veterinary Journal. 2023 May 22; 19(1):
16-23.
[11] Schares G, Barwald A, Staubach C, Wurm R,
Rauser M, Conraths FJ, Schroeder C. Adaptation
of a commercial ELISA for the detection of anti-
bodies against Neospora caninum in bovine milk.
Veterinary Parasitology. 2004 Feb 26; 120(1-2):
55-63.
[12] Meiri-Bendek I, Lipkin E, Friedmann A, Leitner
G, Saran A, Friedman S, Kashi Y. A PCR-based
method for the detectionof Streptococcus agalac-
tiae in milk. Journal of dairy science. 2002 Jul 1;
85(7): 1717-23.
[13] Muller N, Zimmermann V, Hentrich B, Gottstein
B. Diagnosis of Neospora caninum and Toxo-
plasma gondii infection by PCR and DNA hy-
bridization immunoassay. Journal of Clinical Mi-
crobiology. 1996 Nov; 34(11): 2850-2.
[14] Okeoma CM, Williamson NB, Pomroy WE, Stow-
ell KM, Gillespie L. The use of PCR to detect
Neospora caninum DNA in the blood of naturally
infected cows. Veterinary parasitology. 2004 Aug
6; 122(4): 307-15.
[15] Lopez-Perez IC, Risco-Castillo V, Collantes-
Fernandez E, Ortega-Mora LM. Comparative ef-
fect of Neospora caninum infection in BALB/c
mice at three different gestation periods. Journal
of Parasitology. 2006 Dec 1; 92(6): 1286-91.
[16] Gotze A, Honnens A, Flachowsky G, Bollwein
H. Variability of mammary blood ow in lactat-
ing Holstein-Friesian cows during the rst twelve
weeks of lactation. Journal of dairy science. 2010
Jan 1; 93(1): 38-44.
[17] Valasek MA, Repa JJ. The power of real-time
PCR. Advances in physiology education. 2005 Sep;
29(3): 151-9.
[18] Uggla A, Stenlund S, Holmdahl OJ, Jakubek EB,
Thebo P, Kindahl H, Bj'orkman C. Oral Neospora
caninum inoculation of neonatal calves. Interna-
tional Journal for Parasitology. 1998 Sep 1; 28(9):
1467-72.
[19] Davison HC, Guy CS, McGarry JW, Guy F,
Williams DJ, Kelly DF, Trees AJ. Experimental
studies on the transmission of Neospora caninum
between cattle. Research in Veterinary Science.
2001 Apr 1; 70(2): 163-8.
[20] Dijkstra T, Eysker M, Schares G, Conraths FJ,
Wouda W, Barkema HW. Dogs shed Neospora
caninum oocysts after ingestion of naturally in-
fected bovine placenta but not after ingestion of
colostrum spiked with Neospora caninum tachy-
zoites. International journal for parasitology. 2001
Jun 1; 31(8): 747-52.
[21] Lopez AJ, Heinrichs AJ. Invited review: The im-
portance of colostrum in the newborn dairy calf.
Journal of dairy science. 2022 Apr 1; 105(4): 2733-
49.
[22] Vlasova AN, Saif LJ. Bovine immunology: Impli-
cations for dairy cattle. Frontiers in immunology.
2021 Jun 29; 12: 643206.
[23] Taubert A, Zahner H, Hermosilla C. Dynamics
of transcription of immunomodulatory genes in
endothelial cells infected with different coccidian
parasites. Veterinary Parasitology. 2006 Dec 20;
142(3-4): 214-22.
[24] Cohen J. A coefficient of agreement for nominal
scales. Educational and psychological measure-
ment. 1960 Apr; 20(1): 37-46.
last ve years. Veterinary parasitology. 2011 Aug
4; 180(1-2): 90-108.
[2] Wouda W, Moen AR, Schukken YH. Abortion risk
in progeny of cows after a Neospora caninum epi-
demic. Theriogenology. 1998 May 1; 49(7): 1311-6.
[3] Stelwagen K, Carpenter E, Haigh B, Hodgkinson
A, Wheeler TT. Immune components of bovine
colostrum and milk. Journal of animal science.
2009 Apr 1; 87 (suppl 13): 3-9.
[4] Marin MS, Hecker YP, Quintana S, Perez SE,
Leunda MR, Canton GJ, Cobo ER, Moore DP,
Odeon AC. Toll-like receptors 3, 7 and 8 are upreg-
ulated in the placental caruncle and fetal spleen of
Neospora caninum experimentally infected cattle.
Veterinary Parasitology. 2017 Mar 15; 236: 58-61.
[5] Rivera JM, Hecker YP, Burucua MM, Cirone KM,
Cheuquepan FA, Fiorani F, Dorsch MA, Colque
LA, Canton GJ, Marin MS, Moore DP. Innate
and humoral immune parameters at delivery in
colostrum and calves from heifers experimentally
infected with Neospora caninum. Molecular Im-
munology. 2021 Apr 1; 132: 53-9.
[6] Moskwa B, Pastusiak K, Bien J, Cabaj W. The
rst detection of Neospora caninum DNA in the
colostrum of infected cows. Parasitology Research.
2007 Feb; 100: 633-6.
[7] Lindsay DS, Dubey JP. Infections in mice with
tachyzoites and bradyzoites of Neospora caninum
(Protozoa: Apicomplexa). The Journal of para-
sitology. 1990 Jun 1: 410-3.
[8] Moskwa B, Cabaj W. The role of the colostrum
and milk in Neospora caninum transmission.
Helminthologia. 2007 Sep; 44: 126-9.
[9] Mohammed NQ, Aaiz NN. Serological and molec-
ular investigation of Neospora caninum in cattle
in Al-Diwaniyah province, Iraq. Open Veterinary
Journal. 2025 Apr 30; 15(4): 1645.
[10] Ayati R, Rezvan H, Namavari MM, Sadeghi-nasab
A, Hamoon Navard S, Dadmehr B. Association be-
tween presence of DNA and antibody in the serum
during vertical transmission of Neospora caninum.
Iranian Veterinary Journal. 2023 May 22; 19(1):
16-23.
[11] Schares G, Barwald A, Staubach C, Wurm R,
Rauser M, Conraths FJ, Schroeder C. Adaptation
of a commercial ELISA for the detection of anti-
bodies against Neospora caninum in bovine milk.
Veterinary Parasitology. 2004 Feb 26; 120(1-2):
55-63.
[12] Meiri-Bendek I, Lipkin E, Friedmann A, Leitner
G, Saran A, Friedman S, Kashi Y. A PCR-based
method for the detectionof Streptococcus agalac-
tiae in milk. Journal of dairy science. 2002 Jul 1;
85(7): 1717-23.
[13] Muller N, Zimmermann V, Hentrich B, Gottstein
B. Diagnosis of Neospora caninum and Toxo-
plasma gondii infection by PCR and DNA hy-
bridization immunoassay. Journal of Clinical Mi-
crobiology. 1996 Nov; 34(11): 2850-2.
[14] Okeoma CM, Williamson NB, Pomroy WE, Stow-
ell KM, Gillespie L. The use of PCR to detect
Neospora caninum DNA in the blood of naturally
infected cows. Veterinary parasitology. 2004 Aug
6; 122(4): 307-15.
[15] Lopez-Perez IC, Risco-Castillo V, Collantes-
Fernandez E, Ortega-Mora LM. Comparative ef-
fect of Neospora caninum infection in BALB/c
mice at three different gestation periods. Journal
of Parasitology. 2006 Dec 1; 92(6): 1286-91.
[16] Gotze A, Honnens A, Flachowsky G, Bollwein
H. Variability of mammary blood ow in lactat-
ing Holstein-Friesian cows during the rst twelve
weeks of lactation. Journal of dairy science. 2010
Jan 1; 93(1): 38-44.
[17] Valasek MA, Repa JJ. The power of real-time
PCR. Advances in physiology education. 2005 Sep;
29(3): 151-9.
[18] Uggla A, Stenlund S, Holmdahl OJ, Jakubek EB,
Thebo P, Kindahl H, Bj'orkman C. Oral Neospora
caninum inoculation of neonatal calves. Interna-
tional Journal for Parasitology. 1998 Sep 1; 28(9):
1467-72.
[19] Davison HC, Guy CS, McGarry JW, Guy F,
Williams DJ, Kelly DF, Trees AJ. Experimental
studies on the transmission of Neospora caninum
between cattle. Research in Veterinary Science.
2001 Apr 1; 70(2): 163-8.
[20] Dijkstra T, Eysker M, Schares G, Conraths FJ,
Wouda W, Barkema HW. Dogs shed Neospora
caninum oocysts after ingestion of naturally in-
fected bovine placenta but not after ingestion of
colostrum spiked with Neospora caninum tachy-
zoites. International journal for parasitology. 2001
Jun 1; 31(8): 747-52.
[21] Lopez AJ, Heinrichs AJ. Invited review: The im-
portance of colostrum in the newborn dairy calf.
Journal of dairy science. 2022 Apr 1; 105(4): 2733-
49.
[22] Vlasova AN, Saif LJ. Bovine immunology: Impli-
cations for dairy cattle. Frontiers in immunology.
2021 Jun 29; 12: 643206.
[23] Taubert A, Zahner H, Hermosilla C. Dynamics
of transcription of immunomodulatory genes in
endothelial cells infected with different coccidian
parasites. Veterinary Parasitology. 2006 Dec 20;
142(3-4): 214-22.
[24] Cohen J. A coefficient of agreement for nominal
scales. Educational and psychological measure-
ment. 1960 Apr; 20(1): 37-46.
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