




































 

 

 
65 

† Corresponding author 
© 2014 Conscientia Beam. All Rights Reserved. 

IDENTIFICATION OF CATTLE PERSISTENTLY INFECTED WITH BVDV 

(PI) BY EAR-NOTCH TESTING IN SOUTHEAST OF IRAN 

 

Ali Asghar Mozaffari1† --- Mohammad Khalili2 --- Farzaneh Jahangosha3 

1Department of Clinical Studies, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran  

2Department of Pathobiology, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran  

3Graduated, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran 

 

ABSTRACT 

Bovine viral diarrhea virus (BVDV) infection can induce a variety of economically important clinical 

manifestations in cattle herds. One potential outcome is the creation of calves that are viremic but 

immunotolerant to the virus. These persistently infected (PI) calves are the result of in utero exposure to 

BVDV prior to the development of a competent fetal immune system. The aim of this study was to detect of 

cattle persistently infected with BVDV by ear-notch testing in southeast of Iran. Ear-notch skin samples, 3 

mm in diameter, were collected from a total of 127 cattle from 6 randomly selected herds (calves aged under 

12 months ), using pliers usually used for ear tagging and skin notch sampling, as described by the 

manufacturer of the Herd check ELISA kit (IDEXX) for the detection of BVDV antigen in PI cattle. 

Overall, 0.78 per cent of the animals examined, were positive for BVDV antigen.This study identifies the 

cattle persistently infected with BVDV by ear-notch testing in southeast of Iran for the first time. 

Keywords: BVDV, PI, Cattle, Iran, Ear-notch. 

 

Contribution/ Originality 

This study identifies the cattle persistently infected with BVDV by ear-notch testing in 

southeast of Iran for the first time. 

 

1. INTRODUCTION 

BVDV is one of the most economically important pathogens in the cattle industry nowadays 

[1-4]. Bovine viral diarrhea virus (BVDV) infection can produce a variety of economically 

important clinical manifestations in cattle herds. One potential outcome is the creation of calves 

that are viremic but immunotolerant to the virus. These persistently infected (PI) calves are the 

result of in utero exposure to BVDV prior to the development of a competent fetal immune 

system [5]. The calves are persistently viremic and continue to shed the virus for the rest of their 

lives. The neonatal mortality of PI calves is high and some of them are born weak [6]. PI calves 

can present as stunted animals with an unthrifty coat, but not all PI animals are in poor condition 

Animal Review 
2014 Vol. 1, No. 4, pp. 65-68 
ISSN(e): 2409-6490 
ISSN(p): 2412-3382 
© 2014 Conscientia Beam. All Rights Reserved. 
 

 
 



Animal Review, 2014, 1(4): 65-68 

 

 
66 

© 2014 Conscientia Beam. All Rights Reserved. 

and it is not possible to diagnosis them from the physical appearance [7].  The real danger of PI 

calves lies in the fact that they are persistently viremic, immunosuppresed and 

constantly/intermittently shedding virus, and are the main source of infection for other animals 

[8, 9]. If PI calves can be detected, they can be removed in time to prevent spreading of virus to 

susceptible animals [10]. To the best of our knowledge, no report has been published on cattle 

persistently infected with BVDV in Iran. The aim of this study was to detect of cattle persistently 

infected with BVDV by ear-notch testing in southeast of Iran.  

 

2. MATERIALS AND METHODS 

Ear-notch skin samples, 3 mm in diameter, were collected from a total of 127 cattle (Iranian 

cross-breed) from 6 randomly selected herds (dairy cattle with different numbers) in Kerman 

province of Iran (calves aged under 12 months), using pliers usually used for ear tagging and skin 

notch sampling, as described by the manufacturer of the Herd Check ELISA kit (IDEXX) for the 

detection of BVDV antigen in PI cattle. The collected samples were covered with the soaking 

buffer provided in the ELISA kit, and were stored at –80°C until required. The ELISA kit was 

then used on the collected ear-notch samples, according to the manufacturer’s instructions. The 

IDEXX BVDV Ag/Serum plus Test is an enzyme-linked immunoassay for the detection of 

bovine viral diarrhea virus antigen in serum, plasma, whole blood and ear-notch tissue samples. 

To confirm PI, the animals were tested two or three weeks after first sampling. 

 

3. RESULTS  

Table 1 shows the results for detection of BVDV antigen in the 127 ear-notch samples. 

Overall, 0.78 per cent of the animals examined were positive for BVDV antigen. The PI animal 

was not tested for antibodies anti-BVDV. The clinical condition of PI Animals was normal in 

other respect. The history of studied farms showed a number of reproductive/respiratory 

problems, abortion, etc. 

 

4. DISCUSSION  

Ag ELISA using ear-notch samples is an efficient technique for diagnosis of PI calves [11]. 

Ear-notch testing is respected by many researchers as the method of choice to detect PI animals 

in cattle herds, as the samples are easy to collect, sophisticated equipment is not required, the 

samples can be used in a number of BVDV detection systems and they are not affected by the 

presence of passive antibodies [3, 11-15]. 

Results of present study were accordant with records in the literature from other countries 

that indicated that up to 2 percent of cattle are PI with BVDV in most countries [3, 11, 15, 16].  

The results of the this study indicate that there is BVDV activity in the farms under study, 

which necessitates a control policy to be implemented immediately [17, 18].  

Firstly, all existing PI calves should be eliminated from the herd, and routine testing of cattle 

should be introduced to enable the early identification and removal of new PI animals [15].  



Animal Review, 2014, 1(4): 65-68 

 

 
67 

© 2014 Conscientia Beam. All Rights Reserved. 

Secondly, precolostral serum samples from apparently healthy neonatal calves could be tested 

to determine exposure to BVDV in utero from 150 days of gestation; at this gestational age the 

fetus will be able to produce antibodies to BVDV, which will protect it from the ill effects of the 

virus, and the antibodies will be detectable in serum samples taken before colostrum feeding. 

Calves infected in utero before 150 days will not mount an antibody response and can be PI.  

The third, the tissues of neonatal calves showing congenital malformations should be tested 

to confirm their exposure to BVDV.  

At last, surveillance should be carried out on the farm for animals showing clinical signs 

suspicious of mucosal disease [15].  

As a result of present work, it is suggested that other dairy farms in Iran also need to carry 

out a similar program of testing for BVDV and then adopt an appropriate control policy in the 

light of the findings [3, 15]. To the best of our knowledge, no report has been published on 

calves persistently infected with BVDV in Iran. This study identifies the calves persistently 

infected with BVDV by ear-notch testing in southeast of Iran for the first time. 

 

REFERENCES 

[1] I. Firat, S. Ak, and H. Bozkurt, "Distribution of bovine viral diarrhoea virus (BVDV) in the genital 

system tissues of cattle," Vet. Arhiv., vol. 72, pp. 235-248, 2002. 

[2] M. Van Vuuren, "Bovine viral diarrhea virus infection in livestock in Southern Africa. CAB 

reviews," Perspecfives in Agriculture, Veterinary Science, Nutrifion and Natural Resources, vol. 2005, pp. 

4-12, 2005. 

[3] H. Houe, A. Lindberg, and V. Moennig, "Test strategies in bovine viral diarrhea virus control and 

eradication campaigns in Europe," J. Vet. Diagn. Invest., vol. 18, pp. 427-436, 2006. 

[4] M. Hilbe, H. Stalder, and E. Peterhans, "Comparison of five diagnostic methods for detecting 

bovine viral diarrhea virus infection in calves," J. Vet. Diagn. Invest., vol. 19, pp. 28-34, 2007. 

[5] T. Wittum, D. Grotelueschen, and K. Brock, "Persistent bovine viral diarrhoea virus infection in 

US beef herds," Prev. Vet. Med., vol. 49, pp. 83-94, 2001. 

[6] L. A. Moczygemba, "Review of the relationship between persistent infection of cattle with bovine 

viral diarrhea virus and feedlot morbidity and gain," Bovine Pr., vol. 37, pp. 155-161, 2003. 

[7] L. Potgieter, "Bovine viral diarrhoea and mucosal disease," Infect Dis Livestock, vol. 2, pp. 946-969, 

2004. 

[8] J. Kampa, K. Stahl, and L. Renstrom, "Evaluation of a commercial erns-capture ELISA for 

detection of BVDV in routine diagnostic cattle serum samples," Acta Vet. Scand., vol. 49, pp. 1-7, 

2007. 

[9] C. Luzzago, M. Frigerio, and F. Tolari, "Indirect immunohistochemistry on skin biopsy for the 

detection of persistently infected cattle with bovine viral diarrhoea virus in Italian dairy herds," 

New Microbiol., vol. 29, pp. 127-131, 2006. 

[10] T. Meiring, L. Prozesky, E. R. Du Preez, and D. J. Verwoerd, "The diagnosis and prevalence of 

persistent infection with bovine viral diarrhea virus in South African feedlot cattle," Onderstepoort 

J. Vet. Res., vol. 78, p. 323, 2011. 



Animal Review, 2014, 1(4): 65-68 

 

 
68 

© 2014 Conscientia Beam. All Rights Reserved. 

[11] T. Cornish, A. Van Olphen, and J. Cavender, "Comparison of ear notch immunohistochemistry, ear 

notch antigen-capture ELISA, and buffy coat virus isolation for detection of calves persistently 

infected with bovine viral diarrhea virus," J. Vet. Diagn. Invest., vol. 17, pp. 110-117, 2005. 

[12] S. Kuhne, C. Schroeder, and G. Holmquist, "Detection of bovine viral diarrhoea virus infected 

cattleâ€“testing tissue samples derived from ear tagging using an Erns capture ELISA," J. Vet. 

Med. B., vol. 52, pp. 272-277, 2005. 

[13] J. Ridpath, B. Hessman, and J. Neill, "Parameters of ear notch samples for BVDV testing: Stability, 

size requirements and viral load," presented at the Paper Presented at: Proc Am Assoc Bov Pract 

Conf., 2006. 

[14] M. Al-Khaliyfa, E. Abuelzein, and A. Gameel, "Identification of cattle persistently infected with 

BVDV by ear-notch testing in Saudi Arabia," Vet. Rec., vol. 167, pp. 660-661, 2010. 

[15] J. Kampa, Epidemiology of bovine viral diarrhoea virus and bovine herpesvirus type1 infections in dairy 

cattle herds. Uppsala: Sveriges lantbruksuniv, 2006, 2006. 

[16] A. Lindberg and S. Alenius, "Principles for eradication of bovine viral diarrhoea virus (BVDV) 

infections in cattle populations," Vet. Microbiol., vol. 64, pp. 197-222, 1999. 

[17] Components and Goals of Programs to Control BVDV, Available: 

www.ars.usda.gov/SP2UserFiles/Place/36253000/BVD2005/Prod3_Smith_Hout.pdf.UpdatedLa

stUpdatedDate. [Accessed January 8, 2013], 2005. 

[18] P. Roeder and J. Harkness, "BVD virus infection: Prospects for control," Vet. Rec., vol. 119, pp. 143-

147, 1986. 

 

Table-1. Detection by ELISA of BVDV antigen in ear notches from calves in 6 dairy 

farm herds in southeast of Iran 

Animals  Number tested  Number positive (%) 

Herd 1(87) 27 0 
Herd 2(56) 23 1(0.78) 
Herd 3(92) 24 0 
Herd 4(69) 19 0 

Herd 5(55) 18 0 
Herd 6(83) 16 0 
Total (442) 127 1(0.78) 

 

 

 

 

 

 

Views and opinions expressed in this article are the views and opinions of  the author(s), Animal Review shall not be responsible or 
answerable for any loss, damage or liability etc. caused in relation to/arising out of  the use of  the content.  

 

http://www.ars.usda.gov/SP2UserFiles/Place/36253000/BVD2005/Prod3_Smith_Hout.pdf.UpdatedLastUpdatedDate
http://www.ars.usda.gov/SP2UserFiles/Place/36253000/BVD2005/Prod3_Smith_Hout.pdf.UpdatedLastUpdatedDate

