Preventive Veterinary Medicine 175 (2020) 104863

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Preventive Veterinary Medicine

journal homepage: www.elsevier.com/locate/prevetmed

Sero-prevalence and risk factors of infectious bovine rhinotracheitis virus T

(type 1) in Meru County, Kenya
Essau Serem Kipyegoa,*, George Gitaua, John Vanleeuwenb, Peter Kimelia,b,
Tequiero Okumu Abuoma, Daniel Gakuyaa, Joan Murayaa,b, Dennis Makaub
a
University of Nairobi, Nairobi, Kenya
b
University of Prince Edward Island

ARTICLE INFO ABSTRACT

Keywords: The aim of the study was to determine the antibody sero-prevalence of Bovine Herpesvirus-1 which cause
Dairy cattle Infectious Bovine Rhinotracheitis (IBR) and to identify risk factors associated with BHV-1 antibody seropositivity
Infectious Bovine Rhinotracheitis (IBR) among smallholder dairy farms in Meru County, Kenya.
Bovine Herpesvirus type 1 (BHV-1) A cross-sectional study was conducted in the Naari area of Meru County, Kenya between September–October
Sero-prevalence
2016 and March–April 2017. The 149 farmers were randomly selected from members of the Naari Dairy Farmers
Risk factors
Cooperative Society who were actively delivering milk to the society at the time of the study. Serum samples
were obtained from 403 female dairy cattle. Farm level management and animal factors were collected through
direct interviews with the owner or someone who was knowledgeable about the animals. All serum samples were
processed with an indirect enzyme-linked immunosorbent assay (gB ELISA) to determine the presence of anti-
bodies to BHV-1.
The overall farm-level and animal-level sero-prevalences of BHV-1 antibodies were 30.9 % (95 % CI:
23.6%–39.0%) and 17.4 % (95 % CI: 13.8%–21.4%), respectively. In the final multivariable analysis, the factors
significantly associated with BHV-1 antibodies included; age of the dairy cattle (OR = 1.200, p = 0.001), age of
the principal female farmers (OR = 0.182, p = 0.001) and rearing goats in the farm (OR = 26.77, p = 0.000).
There was a significant interaction between rearing goats on the farm and age of the dairy cattle (p < 0.010);
younger cattle seemed to have been exposed to BHV or a cross-reacting caprine herpesvirus when goats were on
the farm.
The results showed that BHV-1 was circulating among the cattle population in the Naari area of Meru County.
Given that there is not BHV-1 vaccination use in this study population, training on the importance of biosecurity
and vaccination for BHV-1 are recommended to reduce the transmission and impacts of BHV-1.

1. Introduction
Infectious Bovine Rhinotracheitis (IBR) is a multi-organ disease of
significant economic importance worldwide caused by Bovine Herpes
Virus-1 (BHV-1), and it affects both domestic and wild ruminants
(Bowland and Shewen, 2000; Muylkens et al., 2007). Bovine Herpes
Virus-1 is a virus of the genus Varicellovirus, subfamily Alphaherpesvir-
inae and family Herpesviridae, and is a highly contagious and infectious
virus (King et al., 2012; Biswas et al., 2013; Newcomer and Givens,
2016). Infectious bovine rhinotracheitis in bovine animals is caused by
BHV-1.1, the respiratory subtypes, while strain BHV-1.2a and BHV-1.2b
are the genital subtypes, and BHV-1.3 is the encephalitic subtype
(Graham, 2013; Muylkens et al., 2007).
Contact with Infected cattle is the main source of BHV-1 infection to
susceptible animals and herds. These cattle shed the virus through
various body secretions and excretions (Takiuchi et al., 2005; Constable
et al., 2017). For the respiratory subtype, the virus is transmitted
through direct transmission of nasal discharges between animals or by
indirect transmission of aerosol droplets, and the infectivity duration of
these aerosols is dependent on environmental factors such as humidity
and temperature. Direct contact with non-nasal mucosal discharges,
contaminated semen, fetal tissues, and genital fluid can also lead to
transmission (Mars et al., 1999, 2000; Kahrs, 2001).
Cattle of all breeds and ages are equally susceptible, and the disease
is common in cattle above 6 months of age due to waning of maternal
antibody protection and increased mixing of cattle populations

⁎
Corresponding author.
E-mail address: kipyegoserem@students.uonbi.ac.ke (E.S. Kipyego).

https://doi.org/10.1016/j.prevetmed.2019.104863
Received 7 July 2019; Received in revised form 16 November 2019; Accepted 29 November 2019
0167-5877/ © 2019 Elsevier B.V. All rights reserved.
E.S. Kipyego, et al.
(Radostits et al., 2000; Majumder et al., 2015; Seyfi Abad Shapouri
et al., 2016; Constable et al., 2017). The IBR diseases have no inherent
seasonal variability, although in temperate countries, during the
months of fall and winter, disease occurrence is high due to feedlot
cattle assembling (Majumder et al., 2015). The managerial and en-
vironmental risk factors contributing to the spread of BHV–1 include;
purchasing of infected cattle, participation in agricultural shows, in-
creased herd size and type of production system (Gay and Barnouin,
2009; Constable et al., 2017). Uncontrolled movement of visitors and
cattle into the farm, and unreliable records of vaccination dates have
also been associated with the spread of the disease (Boelaert et al.,
2005; González-Garcia et al., 2009).
The prevalence and risk factors of BHV-1 infection in Kenya have
not been recently researched in predominantly zero-grazing farming
regions of Kenya. In this cross-sectional study, a random sample of dairy
cattle was examined to determine the BHV-1 antibody sero-prevalence,
and to identify risk factors associated with BHV-1 antibody ser-
opositivity on smallholder dairy farms using primarily zero-grazing in
Naari area, Meru County, Kenya.
2. Materials and methods
2.1. Ethical approval
The study was approved by the Biosafety, Animal use and Ethics
Committee, Faculty of Veterinary Medicine, University of Nairobi
(FVM/BAUEC/2018/146).
2.2. Selection of study area and farms
The study area of Naari in Meru County, Kenya (Fig. 1), was pur-
posively selected since this research formed part of a larger study in-
volving smallholder dairy farmers. A non-governmental organization
called Farmers Helping Farmers, the University of Prince Edward Island
(Canada) and the University of Nairobi had an existing developmental
partnership with the Naari Dairy Farmers Cooperative Society, and this
formed the basis for the entry point to the community.
Meru lies at latitude and longitude 0°6′0″ N and 37°34′60″ E, re-
spectively, and is located in the Mount Kenya region of Kenya, ap-
proximately 270 km North of Nairobi, the capital city of Kenya. The
Naari area is situated at an altitude of approximately 2000 m above sea
Fig. 1. A map showing Naari Sub-location (middle of county) in Meru County of Kenya.
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Preventive Veterinary Medicine 175 (2020) 104863
level. The climate in Meru is warm and temperate with volcanic ash
soils, and therefore is considered to have high agricultural potential.
The average annual temperature in the Meru area ranges from 14 °C to
17 °C in the highlands (where Naari is), while in the lowlands it ranges
from 22 °C to 27 °C. Precipitation in the high altitude Meru areas
average 2200 mm, while low altitude areas average 500 mm. The main
agricultural activities in Naari include; lumbering, horticulture, crop
production and dairy production.
The sampling frame for the study consisted of 568 farmers who were
active members of the Naari Dairy Farmers Cooperative Society
(NDFCS) and delivering milk to the cooperative. The sample size of
dairy cattle sampled in the study was estimated based on a prevalence
of BHV-1 infection of 50 % (for a maximum variance), a precision of 5
% and confidence level of 95 %, giving a required sample size of 385
(Dohoo et al., 2009).
The average number of cattle above 6 months of age per farm had
been established to be approximately 3, thus 149 farms were randomly
selected from the 568 smallholder dairy farms from the registry of ac-
tive members using software-based random number generation.
Therefore, the study population of farms represented 26 % of the
farmers who were members of the NDFCS.
2.3. Data and sample collection
The selected farms were visited between September–October 2016
and March–April 2017, and a questionnaire was administered to cap-
ture farm- and animal-level factors of BHV-1. The data collected in-
cluded collection of animal-level information about milking cows and
heifers on the farm. A systematic scrutiny of written farm records was
also conducted to obtain the age and calving history of the cattle, along
with history of respiratory and reproductive diseases, peri-parturient
conditions, and mastitis cases. Other farm-level management informa-
tion collected included: feed and mineral supplementation, vaccination
status (including use of IBR vaccine), cattle owner attendances to any
dairy husbandry training, herd size, awareness and monitoring of heat
signs, and source of animals with respect to the last 12 months.
In addition, 5 ml of whole blood was collected in plain redtop va-
cutainer tubes via the tail vein of each dairy animal greater than 6
months of age, and stored in an ice box during the day. Each evening,
the blood tubes were taken from the ice box and allowed to clot, and
thereafter, the serum was transferred to Eppendorf® vials. The serum
E.S. Kipyego, et al.
was then frozen at −20 °C and transported on ice to the Hematology
and Biochemistry Laboratory, Department of Clinical Studies, Faculty of
Veterinary Medicine, University of Nairobi, and then stored at −20 °C
until all samples were collected for testing.
2.4. Laboratory analysis
The frozen sera were thawed to room temperature (18–26 °C). The
IBR test kit used was the IDEXX IBR gB X3 Ab Test (IDEXX, Liebefeld-
Bern, Switzerland), comprising of a BHV-1 Antigen Coated Plate and all
the reagents, which was also warmed from its storage temperature of
40C to room temperature. The IDEXX IBR gB X3 Ab Test is an Indirect
Enzyme-Linked Immunosorbent Assay (ELISA) which has been devel-
oped to detect presence of antibodies against IBR (type 1) in individual
bovine plasma, milk and serum samples. Antibody responses induced
by vaccines which contain the glycoprotein B (gB) of BHV-1 are de-
tected as well. This kit has a reported sensitivity of 100 % and speci-
ficity of 95 %. The testing procedure was done following the protocol
described by the manufacturer in Liebefeld-Bern, Switzerland.
The ELISA results were read from a microplate photometer, Mindray
Microplate Reader (MR-96A, Shenzhen Mindray Bio-Medical
Electronics Company Limited), where optical density (OD) was mea-
sured either at a single wavelength of 450 nm [A (450)] or dual wa-
velength of 450 nm and 650 nm [A (450/650) for enhanced sensitivity].
The blocking percentage was calculated by using the absorbance ob-
tained with the test sample and subtracting the absorbance of the ne-
gative control containing no BHV-1-specific antibodies and dividing the
result with the negative control containing no BHV-1-specific anti-
bodies. Interpretation of the results was determined via comparing the
sample blocking percentages to the expected percentages in accordance
with the manufacturer’s test instructions: blocking % < 45 = negative;
45 ≤ blocking % < 55 = suspect; and blocking % ≥ 55 = positive.
The suspects were considered as positive in order to obtain a dichot-
omous outcome. A positive control was used on each plate to confirm
that the procedures were followed properly.
2.5. Data entry and statistical analysis
Data collected through the questionnaires and the laboratory results
were first entered into MS Excel (Microsoft Inc., Sacramento, California,
USA) and then imported to Stata 15 (StataCorp LLC, College station,
Texas, USA) for analyses. Initially, the data were checked for accuracy,
coded and analyzed using descriptive statistics. Proportions were de-
termined for categorical variables and presented as a percentage of the
overall number, along with a 95 % confidence interval, where applic-
able.
Mixed-effect logistic regression analysis was performed, accounting
for clustering of cattle among herds, to determine associations between
the categorical and continuous variables and the dichotomized ser-
opositivity outcome (presence or absence of BHV-1 antibody). In the
first step, univariable multi-level mixed models for all the predictor
variables were fitted into separate logistic regression models, em-
ploying the functional logit. In the second step, a multivariable mixed
logistic regression analysis was fitted for all the univariable associations
with p ≤ 0.30 in the first step. Correlations between predictor variables
were identified using pair-wise correlation, and where two or more
variables were highly correlated (correlation coefficient > 0.5), statis-
tical significance and biological plausibility were used to identify which
variable would be offered to the modeling process. The final models
were built using backward stepwise elimination, leaving those variables
which had a p-value ≤0.05. Explanatory variables were considered
confounders if their removal from the multivariable model modified the
coefficients of other significant variables by 30 %. Plausible biological
interactions between significant explanatory variables in final model
were also tested and the significant interaction terms were included in
the final models (Dohoo et al., 2009). The area under the curve (AUC)
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Preventive Veterinary Medicine 175 (2020) 104863
of the receiver operating characteristic was used to evaluate the overall
model performance.
3. Results
3.1. Farm and animal demographics
A total of 403 cattle and 149 farms were involved in the study. The
principal farmer was mostly comprised of men (56.4 %), while women
as principal farmers in this sample population were fewer (28.2 %), and
15.4 % of farms had both the male and female considered jointly as the
principal farmer. Most of the principal farmers were married (83.9 %),
some had lost their spouse, and a few of them were young people who
were still single but had established themselves as dairy farmers (5.3
%). Among the principal farmers, approximately half (50.3 %) were
below 45 years. A large majority of the female farmers had completed
primary school (91.2 %) but only 39.4 % had completed secondary
school, while the proportions of male farmers having completed pri-
mary and secondary school were slightly different at 81.5 % and 42.3
%, respectively. The majority (81.2 %) of principal farmers had training
on dairy production.
The mean household size recorded in this study was 3.7 ± 1.54,
with a minimum of 1 person and a maximum of 11. The mean total land
holdings owned by the respondents was 2.11 ± 2.04 acres. In addition,
some of the farmers also had access to other pieces of land
(0.51 ± 0.84 acres) through renting, borrowing and government-
owned lands leased to them.
Holstein-Friesians formed nearly half of the dairy cattle reared in
the region, with small proportions of other exotic and indigenous
breeds (Table 1). The mean herd size among the sampled farms was 5.7
with a range of 1–16 cattle. The mean number of milking cows was 1.8
with a range of 0 and 8. The mean number of dry cows was 0.3 with a
range of 0–3. Mature cows comprised the majority of animals at 79.7 %
(321/403), while unbred heifers comprised 12.2 % (49/403). The mean
age of the sampled animals was 5.5 years with a range of between 0.5
and 17.0 years. The parities of the cows ranged between 0 and 8, with
multiparous cows forming nearly 60 % of sampled cattle. It was re-
ported that 9.0 % (32/354) of the cows and bred heifers had experi-
enced an abortion in the last one year.
3.2. Farm management practices at the farms
The managerial practices found among the 149 smallholder dairy
farms recruited in the study are summarized in Table 1. The zero-
grazing and semi-zero-grazing systems formed the majority (nearly 80
%) of smallholder dairy farms. Other than dairy production, many of
the recruited farmers also practiced sheep and goat production systems.
The majority of smallholder dairy farms used artificial insemination
exclusively at 57.7 % (86/149). More than half of the farmers grazed
their cows in community pastures where they could contact other
cattle, while less than a quarter of farms disallowed fence-line contact
with other cattle. However, there was insignificant movement of the
cattle between farms across the region, with less than 10 % of farms
borrowing cows, lending cows, and purchasing cows. None of the study
farmers used a BHV-1 vaccine in the past.
3.3. Sero-prevalence of Bovine Herpesvirus-1 infection
The overall apparent animal-level sero-prevalence to BHV-1 was
17.4 % (70/403), with a 95 % CI of 13.8%–21.4%. The overall esti-
mated true animal-level sero-prevalence to BHV-1, adjusting for the
imperfect specificity of the test, was calculated to be 13.0 %, with a 95
% CI of 9.5%–17.2%. The herd-level sero-prevalence to BHV-1 was 30.9
% (46/149), with a 95 % CI of 23.6%–39.0%. The sero-prevalences of
the antibodies to the IBR virus in relation to a number of cow and farm
variables are summarized in Table 1.
E.S. Kipyego, et al. Preventive Veterinary Medicine 175 (2020) 104863

The sero-prevalence for BHV-1 infection was highest among cattle Table 2
that were older and multiparous, with slightly elevated sero-pre- Univariable logistic mixed models of the outcome variable bovine herpesvirus-1
valences among Ayrshire crosses and animals with a history of abortion. antibody seropositivity, while accounting for clustering of 403 cattle among
At the farm level, farms with goats had a 3 times higher sero-prevalence 149 smallholder dairy farms in Meru County, Kenya in 2017, for variables of
interest with P–Value ≤ 0.3.
for BHV-1 among the cattle versus farms without goats, while farms
with sheep only had a slightly higher BHV-1 sero-prevalence in cattle. Variable Categories OR 95 % CIOR P–value
The cattle sero-prevalence for BHV-1 was also substantially higher on
Animal level variables
farms that lent and/or borrowed cattle to/from other farms, and on
Parity category Parity 0 baseline 0.206a
farms where fence-line contact with neighbour’s cattle was allowed. Parity 1 1.059 0.361–3.103 0.917
Parity 2–8 1.414 0.803–4.428 0.145
Age category Heifer baseline
Cows 2.181 0.913–5.211 0.079
Table 1
Age of the dairy cattle n/a 1.112 1.012–1.222 0.027
Description of categorical variables for animal- and farm-level factors for 403
Dry cow status No baseline
cattle on 149 smallholder dairy farms in Meru County, Kenya in 2017, along Yes 1.446 0.685–3.049 0.249
with bovine herpesvirus-1 infection percentages by category. BVDV ab status positiveb Negative baseline
Positive 2.262 1.129–4.533 0.021
Variable Category Category BHV-1 #
Herd level variables
Frequency (Percent) + by
Fence-line contact with other No baseline
(Percent) Category
cattle Yes 2.850 0.924–8.790 0.068
Animal level factors Grazing on community No baseline
Breed Ayrshire 70 (17.4) 14 (20.0) pasture Yes 2.150 0.918–5.033 0.078
Friesian 190 (47.2) 31 (16.3) Borrowed cows from other No baseline
Guernsey 112 (27.8) 20 (17.9) farms
Zebu 31 (7.7) 5 (16.1) Yes 4.893 1.328–16.031 0.017
Age category Cow 321 (79.7) 60 (18.7) Lent cows to other farms No baseline
Heifer 82 (20.3) 10 (12.2) Yes 2.486 0.697–8.859 0.014
Parity category Parity 0 80 (19.9) 12 (15.0) Rearing goats on the farm No baseline
Parity1 86 (21.3) 11 (12.8) Yes 3.024 1.694–5.396 0.001
Parity > 1 237 (58.8) 47 (19.8) Rearing sheep on the farm No baseline
History of an abortion No history of 322 (79.9) 55 (17.1) Yes 1.076 0.969–1.192 0.173
abortion Use of natural mating No baseline
History of 32 (9.0) 15 (18.5) Yes 1.706 0.769–3.786 0.150
abortion Dichotomized age of the < 45 years baseline
BVDV ab positivea Positive 137 (46.6) 32 (23.4) female principal farmer
Negative 157 (53.4) 20 (12.7) > 45 years 0.230 0.108–0.498 0.001
Showed BVDV signs b
Positive 254 (63.2) 41 (16.1) Dichotomized age of the male < 45 years baseline
Negative 148 (36.8) 29 (19.6) principal farmer
Herd level factors > 45 years 0.394 0.181–0.898 0.019
Type of feeding system Zero– grazing 63 (42.2) 11 (17.5)
Semi-zero- 53 (35.6) 10 (18.9) OR: Odds Ratio.
grazing 95 % CIOR: 95 % Confidence Interval of OR.
a
Grazing 33 (22.2) 4 (12.1) Overall P-values for categorical variables with > 2 categories.
Rearing goats on the farm Yes 23 (15.4) 9 (39.1) b
Bovine Viral Diarrhea Disease Virus antibody positive cattle.
No 126 (84.6) 17 (13.5)
Rearing sheep on the farm Yes 57 (38.3) 11 (19.3)
No 92 (61.7) 15 (16.3)
grazed on community pasture, lent cows to other farms, borrowed cows
Use of natural mating Yes 63 (42.3) 12 (19.0)
No 86 (57.7) 14 (16.3) from other farms, reared goats on the farm, reared sheep on the farm,
Fence-line contact with Yes 120 (80.5) 23 (19.2) used natural breeding, age of the male principal farmer, and age of the
other cattle No 29 (19.5) 3 (10.3) female principal farmer (Table 2).
Grazing on community Yes 98 (65.8) 19 (19.4) Pair-wise correlation was carried out and age, parity and age cate-
pasture No 51 (34.2) 7 (13.7)
Borrowed cows from other Yes 10 (6.7) 5 (50.0)
gory of the cattle were found to be highly correlated (r > 0.5). Fence-
farms line contact with other cattle and grazing on community pasture were
No 139 (93.3) 21 (15.1) also found to be highly correlated. Therefore, age of the animal and
Lent cows to other farms Yes 12 (8.1) 4 (33.3) fence-line contact were offered to the final multivariable model because
No 137 (91.9) 22 (16.1)
they had lower P-Values than their correlates.
Cattle bought into the farm Yes 13 (8.7) 3 (23.1)
No 136 (91.3) 23 (16.9) The factors associated with sero-positivity to BHV-1 infection in the
Dichotomized age of the ≤45 years 74 (49.7) 14 (23.9) final multivariable analysis included (Table 3): dichotomized age of the
female principal > 45 years 75 (50.3) 4 (5.3) principal female farmers, age of the dairy cattle reared on the farm, and
farmers rearing goats on the farm. In this final model, controlling for con-
Dichotomized age of the ≤45 years 76 (51.0) 12 (15.8)
male principal farmers > 45 years 73 (49.0) 6 (8.2)
founding of variables in the model, the odds of older dairy cattle to
have BHV-1 antibody positivity were 1.200 times higher with each
a
Bovine Viral Diarrhea Disease Virus antibodies-positive cattle. additional year of age. Cattle on farms that were rearing goats had
b
Bovine Viral Diarrhea disease. 26.77 times the odds to have BHV-1 antibodies compared to cattle on
farms not rearing goats. In addition, when the age of the female prin-
3.4. Factors associated with Bovine Herpesvirus-1 sero-positivity cipal farmer was over 45 years, the odds of cattle on the farm to have
BHV-1 antibodies were lower by 0.182 times compared with cattle
The following animal-level variables met the P–value cut-off cri- reared by younger women principal farmers under or equal to 45 years
terion (P-value < 0.3) for univariable regression analysis with BHV-1 (Table 3). The area under the ROC curve for this final model was 0.86,
antibody status: parity category, age category, dry cow status, and indicating a good overall goodness-of–fit of the observed data (Fig. 2).
BVDV antibody status. Farm-level variables meeting the cut-off cri- The 95 % CI for 1` year and 6 years do not overlap, showing that the
terion included: the farm allowed fence-line contact with other cattle, interaction is significant (p < 0. 0.010). The relationship between

4
E.S. Kipyego, et al.
Table 3
Final multivariable logistic mixed model for variables associated with bovine herpesvirus-1 infection antibody sero-positivity for 403 cattle on 149
smallholder dairy farms in Meru County, Kenya in 2017.
Variable OR
Rearing goats on the farm 26.77
Age of the dairy cattle 1.200
Rearing goats on the farm * Age of the dairy cattle 0.744
Dichotomized age of female principal farmer 0.182
OR: Odd Ratio.
95 % CIOR: 95 % Confidence Interval of OR.
Fig. 2. Area-under-the-curve graph showing goodness-of-fit of the final model
in a study on risk factors for infection with Bovine Herpesvirus-1 for 403 cattle
on 149 smallholder dairy farms in Meru County, Kenya in 2017.
Fig. 3. Interaction plot of the marginal predicted probability and 95 % con-
fidence interval bars of BHV test positivity for age of dairy cattle by goats on
(circle) and off (square) the farms, based on the final model on 403 cattle on
149 smallholder dairy farms in Meru County, Kenya in 2017.
cattle age and BHV antibodies depends on whether there are goats or
not, with there being a significant positive association when the cattle
are young (< 6 years) but no association when the cattle are older than
6 years. Similarly, the relationship between goats on the farm and BHV
antibodies depends on the cattle age, with there being a positive as-
sociation between BHV antibodies and the presence of goats, but only
when cattle age is < 6 years (Fig. 3).
4. Discussion
The results of this study confirmed the presence of IBR infection
through sero-prevalence of BHV-1 antibodies among the smallholder
dairy cattle reared in primarily zero-grazing units in Naari area of Meru
County. The observed sero-prevalence for BHV-1 antibodies from this
5
Preventive Veterinary Medicine 175 (2020) 104863
95 % CIOR P–value
5.328 – 134.5 < 0.001
1.079 – 1.335 0.001
0.593 – 0.933 0.010
0.087 – 0.382 0.001
study was estimated at 17.4 %, which was slightly lower compared to
studies in other parts of Kenya of 20.9 % in the western Kenya (Callaby
et al., 2016) and 28.0 % in the former Malindi District of the Coastal
Region (Kenyanjui et al., 2007). This slightly lower sero-prevalence
could have been due to the different livestock production systems stu-
died, for example, 42 % of our study farms used only zero-grazing, and
open grazing is a known risk factor for BHV–1 infection (Snowder et al.,
2006; Callaby et al., 2016).
The observed sero-prevalence to BHV-1 of 17.4 % from this study
was within the range of 16 %–54 % that was observed by McDermott
et al. (1997) in former Districts across Kenya in 1991–1992. However,
in this study, the suspects were considered as positive in order to obtain
a dichotomous outcome, and this interpretation might have led to
higher magnitudes and direction of estimates of the sero-prevalence
(Friesen et al., 2007 and Dohoo et al., 2009). Conversely, the sero-
prevalence observed in Naari area of Meru County was clearly lower
than that estimated in traditionally managed herds (i.e. open grazing)
in Zambia, which ranged from 42 %–76 % (Straub, 1990; Ghirotti et al.,
1991), and in Egypt which ranged from 63 %–86 % (Mahmoud et al.,
2009).
The observed differences in antibody sero-prevalence in different
regions and countries could be explained by factors such as production
systems, differences in herd sizes, type of breeding methods, differences
in disease-control measures, and age of the cattle (Mainar-Jaime et al.,
2001; Ackermann and Engels, 2006). It has been observed that higher
BHV-1 prevalences recorded in larger herd sizes and intensively farmed
cattle could be associated with a high level of contact between in-
dividual animals within a herd (Snowder et al., 2006). For extensively
managed farms with median herd size of 5 animals, the risk of contact
between a susceptible individual with an infected or persistently in-
fected animal is lower (Callaby et al., 2016). The studies reported above
involved animals of various ages, such as 51 weeks old (Callaby et al.,
2016), 3 months to adults (Ghirotti et al., 1991), and zebu adults
(Kenyanjui et al., 2007), while our study involved heifer and adult dairy
cows. Therefore, the age differences among the animals studied may
also have explained some of the variations in the sero-prevalence.
Ghirotti et al. (1991) and Kenyanjui et al. (2007) employed virus
neutralization tests (VNT) compared to ELISA tests used in our study,
and the differences in test specificity and sensitivity may also have also
contributed to observed differences in prevalence (Graham et al.,
1998). In the studies done by Saravanajayam et al. (2015) it was re-
ported that the ELISA had a higher specificity and sensitivity compared
to VNT, thus ELISA is considered a rapid, reliable and technically su-
perior test for detection of BHV-1 antibodies. Callaby et al. (2016) also
employed an indirect ELISA test, and thus may have contributed a si-
milar prevalence to our study.
The results of the final model of this study revealed that a number of
risk factors were associated with sero-prevalence of BHV-1 antibody,
such as age of the cattle, rearing goats in the farms, and cows that had
antibodies against BVDV. Several serological studies carried out
worldwide identified similar risk factors associated with BHV–1 sero-
positivity (Mars et al., 2001; Muylkens et al., 2007; Callaby et al., 2016;
Constable et al., 2017). The current study also agrees with other studies
which reported a number of risk factors in their studies, such as large
E.S. Kipyego, et al.
herd size, intensive production systems, housing, and management
practices, like animal age and sex (males are more frequently positive
than females), and direct animal contact through cattle shows and
purchasing of cattle (van Schaik, 2001; van Schaik et al., 2002; Solis-
Calderon et al., 2003; Vonk-Noordegraaf et al., 2004; Boelaert et al.,
2005).
Age of the cattle is a frequent risk factor reported in BHV-1 ser-
opositive cattle, where findings from Solis-Calderon et al. (2003),
Carbonero et al.2011, Romero-Salas et al. (2013), Saravanajayam et al.
(2015) and Segura-Correa et al. (2016) showed that older animals had a
higher sero-prevalence compared to young animals. Since infected
cattle remain infected for life, it is not surprising that proportions of
infected cattle increase with age.
Cattle on farms with older female principal farmers had a lower
odds of infection compared to cattle on farms with younger female
principal farmers. It is unclear why this variable remained significant in
our final model, but it may be that older farmers are more knowl-
edgeable of the importance of biosecurity and disease control measures.
Further research would help to clarify this finding.
Based on past epidemiological studies, this study showed that IBR
and BVD viruses are closely associated in the univariable analyses, and
infection for one disease has predisposed to the other disease elsewhere
(Callaby et al., 2016). The study also showed that BVDV sero-pre-
valence was significantly associated with BHV-1 at the farm level. This
finding agrees with those reported earlier (Fulton et al., 2000; Callaby
et al., 2016). A positive correlation has also been demonstrated be-
tween infections with the viruses causing IBR and PI (Callaby et al.,
2016).
The present study revealed that rearing of goats in the farm was
associated with BHV-1 sero-prevalence in the cattle. Mixed rearing of
sheep, goats and cattle was not uncommon in our study population,
allowing this variable to become part of the risk factors in the final
model. Infectious Bovine Rhinotracheitis has the potential for cross-
reaction with four other herpesviruses from other animals including
goats and buffaloes (Handel et al., 2011; Callaby et al., 2016). There-
fore, this risk factor may be attributed to the potential ability of viral
cross-infection with related herpesvirus (Yesilbag et al., 2003; Keuser
et al., 2004; Biswas et al., 2013). Further research is needed to clarify
this potential cross-reaction.
The present study showed that introduction of new animals into the
herd was significantly associated sero-positivity of BHV-1 in univariable
analyses but did not remain significant in the final model. Other past
studies by Solis-Calderon et al. (2003) and Segura-Correa et al. (2016)
reported that introduction of an animal into the herd was not sig-
nificantly associated with sero-prevalence of BHV-1. However, van
Schaik et al. (1998) and Gay and Barnouin (2009) reported increased
animal movement into a herd and close proximity neighbouring farms
increased the risk of IBR spread through contact between naïve herd
and infected animals. While introduction of new animals would biolo-
gically make sense as a risk factor for BHV-1, the relatively low sero-
prevalence of BHV-1 in our study population (13.0 % estimated true
prevalence) and infrequent movement of cattle between herds (6.7 %,
8.1 %, and 8.7 % of farmers borrowed, lent, or bought cattle, respec-
tively – Table 1) would mean that the chances of an infected animal
moving between farms is low.
For the observed significant interaction between cattle age and
goats on the farm (p < 0.01), younger cattle seemed to have been
exposed to BHV or a cross-reacting caprine herpesvirus when goats
were on the farm. This could be because farms with cattle < 6 years old
just brought in goats to the farm in the last 6 years. It could also be that
farms bringing in goats into the farm in the last 6 years is correlated
with farms bringing in other cattle in the last 6 years, and so those
younger cattle are more likely to be exposed to BHV with these new
cattle introductions than older cattle.
This study was carried out in smallholder farms with approximate
herd size of 3 animals and little variation in herd size, as a result, the
6
Preventive Veterinary Medicine 175 (2020) 104863
study did not have enough variation and power to identify herd size as a
risk factor for seropositivity. Other studies suggested that at herd level,
herd size is the main important risk factor for BHV-1 infection (Snowder
et al., 2006; Segura-Correa et al., 2016). Our herd sizes were small,
making it less likely for our herds to be aggregates of animals from
other farms.
A limitation to this study is that it is a cross-sectional design, which
has limited ability to confirm temporality of the risk factors for BHV-1
infection. Furthermore, the study used a BHV-1-specific antibody ELISA
testing kit, thus this test cannot establish whether the test-positive an-
imal is due to the virus retention, or due to adaptive immunity from
previous exposure to the infection. In the future, a study is needed to
establish if clinical IBR occurs in smallholder dairy farmers in the Meru
area of Kenya in order to determine the long-term effects of BHV-1
infection in cattle production that would justify formulation of pre-
vention and control measures, such as vaccination.
5. Conclusions and recommendations
Overall, BHV-1 was found to be naturally circulating among the
cattle population in Meru County. There was a positive association
between BHV-1 infection and cattle age, rearing of goats in the farm
together with cattle, and younger women principal farmers. Given that
there is not BHV-1 vaccination use in this study population, training on
the importance of biosecurity and vaccination for BHV-1 are re-
commended to reduce the transmission and impacts of BHV-1.
Declaration of Competing Interest
None.
Acknowledgements
We are grateful to the primary funding program for this research,
the Canadian Queen Elizabeth II Diamond Jubilee Scholarships which
are managed through a unique partnership of universities Canada, the
Rideau Hall Foundation, Community Foundations of Canada and
Canadian universities. This program is made possible with financial
support from the Government of Canada, provincial governments and
the private sector. We also acknowledge the large contribution made by
University of Nairobi, volunteers and staff of Farmers Helping Farmers,
a non-governmental organization – their existing relationships and
agricultural efforts and inputs provided a strong foundation for the
work and the entry point to the Naari community. As well, the support
of the Naari Dairy Farmers Cooperative Society.
References
Ackermann, M., Engels, M., 2006. Pro and contra IBR-eradication. Vet. Microbiol. 113
(3–4), 293–302. https://doi.org/10.1016/j.vetmic.2005.11.043.
Biswas, S., Bandyopadhyay, S., Dimri, U., Patra, H.P., 2013. Bovine herpesvirus-1 (BHV-
1) – a re-emerging concern in livestock: a revisit to its biology, epidemiology, diag-
nosis, and prophylaxis. Vet. Q. 33 (2), 68–81. https://doi.org/10.1080/01652176.
2013.799301.
Boelaert, F., Speybroeck, N., De Kruif, A., Aerts, M., Burzykowski, T., Molenberghs, G.,
Berkvens, D.L., 2005. Risk factors for bovine herpesvirus-1 seropositivity. Prev. Vet.
Med. 69, 285–295. https://doi.org/10.1016/j.prevetmed.2005.02.010.
Bowland, S.L., Shewen, P.E., 2000. Bovine respiratory disease: commercial vaccines
currently available in Canada. Can. Vet. J. 41, 33–48. https://www.ncbi.nlm.nih.
gov/pmc/articles/PMC1476343/.
Callaby, R., Toye, P.G., Jennings, A., Thumbi, S.M., Coveter, J.A.W., Van Wyk, I.C.,
Hanotte, O., Mbole-Kariuki, M.N., Bronsvoort, M.D.C., Kruuk, L.E.B., Woolhouse,
M.E.J., Kiara, H., 2016. Sero-prevalence of respiratory viral pathogens of indigenous
calves in Western Kenya. Res. Vet. Sci. 108, 120–124. https://doi.org/10.1016/j.rvsc.
2016.08.010./.
Carbonero, A., Sa a, L.R., Jara, D.V., García-Bocanegra, I., Arenas, A., Borge, C., Perea, A.,
2011. Sero-prevalence and risk factors associated to Bovine Herpesvirus 1 (BHV-1)
infection in non-vaccinated dairy and dual purpose cattle herds in Ecuador. Prev. Vet.
Med. 100 (1), 84–88. https://doi.org/10.1016/j.prevetmed.2011.03.006.
Constable, P.D., Hinchcliff, K.W., Done, S.H., Grünberg, W., 2017. Infectious Bovine
Rhinotracheitis. Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses,
E.S. Kipyego, et al.
Sheep, Pigs, and Goats, 11th ed. Elsevier, St. Louis, Missouri, pp. 952–961.
Dohoo, I.R., Martin, W., Stryhn, H., 2009. Veterinary Epidemiologic Research, 2nd ed.
AVC Inc., Charlottetown, PEI, Canada.
Friesen, M.C., Davies, H.W., Teschke, K., Ostry, A.S., Hertzman, C., Demers, P.A., 2007.
Impact of the specificity of the exposure metric on exposure-response relationships.
Epidemiology 18, 88–94. https://doi.org/10.1097/01.ede.0000249558.18960.6b.
Fulton, R.W., Purdy, C.W., Confer, A.W., Saliki, J.T., Loan, R.W., Briggs, R.E., Burge, L.J.,
2000. Bovine viral diarrhea viral infections in feeder calves with respiratory disease:
interactions with Pasteurella spp., parainfluenza-3 virus, and bovine respiratory
syncytial virus. Can. J. Vet. Res. 64, 151–159. https://www.ncbi.nlm.nih.gov/pmc/
articles/PMC1189606/.
Gay, E., Barnouin, J., 2009. A nation-wide epidemiological study of acute bovine re-
spiratory disease in France. Prev. Vet. Med. 89, 265–271. https://doi.org/10.1016/j.
prevetmed.2009.02.013.
Ghirotti, M., Semproni, G., Meneghi, D., Mungaba, F.N., Nannini, D., Calzetta, G.,
Paganico, G., 1991. Sero-prevalence’s of selected cattle diseases in the Kafue flats of
Zambia. Vet. Res. Commun. 15, 25–36. https://link.springer.com/article/10.1007/
BF00497787.
González-Garcia, M.A., Arenas-Casas, A., Carbonero-Martínez, A., Borge-Rodríguez, C.,
García-Bocanegra, I., Maldonado, J.L., Gómez, J.M., Perea-Remujo, J.A., 2009.
Seroprevalence and risk factors associated with bovine herpesvirus type 1 (BHV-1)
infection in non-vaccinated cattle herds in Andalusia (South of Spain). Spanish J.
Agric. Res. 3, 550–554. https://helvia.uco.es/handle/10396/11847.
Graham, D.A., 2013. Bovine herpes virus – 1 (BHV – 1) in cattle – a review with emphasis
on reproductive impacts and emergence of infection in Ireland and United Kingdom.
Ir. Vet. J. 66, 15. https://doi.org/10.1186/2046-0481-66-15.
Graham, D., McShane, J., Mawhinney, K., McLaren, I., Adair, B., Merza, M., 1998.
Evaluation of a single dilution ELISA system for detection of seroconversion to bovine
viral diarrhea virus, bovine respiratory syncytial virus, parainfluenza-3 virus, and
infectious bovine rhinotracheitis virus: comparison with testing by virus neutraliza-
tion and hemagglutination inhibition. J. Vet. Diagn. Invest. 10, 43–48. https://doi.
org/10.1177/104063879801000108.
Handel, I.G., Willoughby, K., Land, F., Koterwas, B., Morgan, K.L., Tanya, V.N., Barend,
M., 2011. Seroepidemiology of bovine viral diarrhea virus (BVDV) in the Adamawa
region of Cameroon and use of the SPOT test to identify herds with PI calves. PLoS
One 6 (7), e21620. https://doi.org/10.1371/journal.pone.0021620.
Kahrs, R.F., 2001. Infectious bovine rhinotracheitis and infections pustular vulvovaginitis.
Viral Diseases of Cattle, 2nd ed. Iowa State University Press, Ames, IA, pp. 159–170.
https://doi.org/10.1080/01652176.2013.799301.
Kenyanjui, M., Steiger, Y., Thorpe, W., 2007. Virus neutralizing Anitbodies to bovine
herpes virus type 1 (BHV-1), bovine viral diarrhoea (BVD) and rinderpest (RPV)
viruses in smallholder east African zebu cattle in coastal Kenya. Kenya Veterinarians
18, 21–24. https://www.ajol.info/index.php/kenvet/article/view/39494.
Keuser, V., Schynts, F., Detry, B., Collard, A., Robert, B., Vanderplasschen, A., Pastoret, P.,
Thirty, E., 2004. Improved antigenic methods for differential of bovine diagnosis of
bovine, caprine, and corvine alpha-herpesviruses related to bovine herpesvirus-1.
Clinical Microbiology 42, 1228–1235. https://doi.org/10.1128/JCM.42.3.1228-
1235.2004.
King, A.M.Q., Adams, M.J., Carstens, E.B., Lefkowitz, E.J., 2012. Virus Taxonomy:
Classification and Nomenclature of Viruses: Ninth Report of the International
Committee on Taxonomy of Viruses. CA: Elsevier/Academic Press, San Diego, pp.
1010.
Mahmoud, M.A., Mahmoud, N.A., Allam, A.M., 2009. Investigation on infectious bovine
rhinotracheitis in Egyptian cattle and buffaloes. Glob. Vet. 3, 335–340.
Mainar-Jaime, R.C., Berzal-Herranz, B., Arias, P., Rojo-Vázquez, F.A., 2001.
Epidemiological pattern and risk factors associated with bovine viral-diarrhoea virus
(BVDV) infection in a non-vaccinated dairy-cattle population from the Asturias re-
gion of Spain. Prev. Vet. Med. 52 (1), 63–73. https://doi.org/10.1016/S0167-
5877(01)00239-2.
Majumder, S., Ramakrishnan, M.A., Nandi, S., 2015. Infectious bovine rhinotracheitis: an
indian perspective. Int. J. Curr. Microbiol. Appl. Sci. 4 (10), 844–858.
Mars, M.H., de Jong, M., Franken, P., van Oirschot, J.T., 2001. Efficacy of a live glyco-
protein E-negative bovine herpesvirus 1 vaccine in cattle in the field. Vaccine 19,
1924–1930. https://doi.org/10.1016/S0264-410X(00)00435-7.
Preventive Veterinary Medicine 175 (2020) 104863
Mars, M.H., De Jong, M.C., Van-Maanen, C., Hage, J.J., Van Oirschot, J.T., 2000.
Airborne transmission of bovine herpesvirus 1 infections in calves under field con-
ditions. Vet. Microbiol. 76, 1–13. https://doi.org/10.1016/S0378-1135(00)00218-2.
Mars, M.H., Bruschke, C.J., van Oirschot, J.T., 1999. Airborne transmission of BHV1,
BRSV, and BVDV among cattle is possible under experimental conditions. Vet.
Microbiol. 66 (3), 197–207. https://doi.org/10.1016/S0378-1135(99)00009-7.
McDermott, J.J., Kadohira, M., O’Callaghan, C.J., Shoukri, M.M., 1997. A comparison of
different models for assessing variations in the sero-prevalence of infectious bovine
rhinotracheitis by farm, area and district in Kenya. Prev. Vet. Med. 32 (3–4),
219–234. https://doi.org/10.1016/S0167-5877(97)00025-1.
Muylkens, B., Thiry, J., Kirten, P., Schynts, F., Thiry, E., 2007. Bovine herpesvirus 1 in-
fection and infectious bovine rhinotracheitis. Vet. Rec. 38, 181–209. https://doi.org/
10.1051/vetres:2006059.2007.181.209. https://doi.org/10.1051/vetres:2006059.
Newcomer, B.W., Givens, D., 2016. Diagnosis and control of viral diseases of reproductive
importance: infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea. Vet. Clin.
N. Am-Food A 34, 425–441. https://doi.org/10.1016/j.cvfa.2016.01.011.
Radostits, O.M., Gay, C.C., Blood, D.C., Hinchcliff, K.W., 2000. Infectious Bovine
Rhinotracheitis (IBR, Red Nose), Bovine Herpesvirus -1 (BHV-1) Infection, 9th ed.
W.B. Saunders, London. UK, pp. 1173–1184.
Romero-Salas, D., Ahuja-Aguirre, C., Montiel-Palacios, F., García-Vázquez, Z., Cruz-
Romero, A., Aguilar-Domínguez, M., 2013. Seroprevalence and risk factors associated
with infectious bovine rhinotracheitis in unvaccinated cattle in southern Veracruz,
Mexico. Afr. J. Microbiol. Res. 7 (17), 1716–1722. https://doi.org/10.5897/AJMR12.
1334.
Saravanajayam, M., Kumanan, K., Balasubramaniam, A., 2015. Seroepidemiology of in-
fectious bovine rhinotracheitis infection in unvaccinated cattle. Vet. World 8 (12),
1416–1419. https://doi.org/10.14202/vetworld.2015.1416-1419.
Segura-Correa, J.C., Zapata-Campos, C.C., Jasso-Obregón, J.O., Martinez-Burnes, J.,
López-Zavala, R., 2016. Sero-prevalence and risk factors associated with bovine
herpesvirus 1 and bovine viral diarrhea virus in North-Eastern Mexico. Open Vet. J. 6
(2), 143–149. https://doi.org/10.4314/ovj.v6i2.12.
Seyfi Abad Shapouri, M., Ghiami, R.M., Haji Hajikolaei, M.R., Mahmoodi, P, Karami, A.,
Daghari, M., 2016. Isolation of bovine Herpesvirus-1 (BoHV-1) from latently
Infected/Carrier cattle in Ahvaz. Iran. J. Rumi. Health Res. 1 (1), 11–20.
Solis-Calderon, J.J., Segura-Correa, V.M., Segura-Correa, J.C., Alvarado-Islas, A., 2003.
Sero-prevalence of and risk factors for infectious bovine rhinotracheitis in beef cattle
herds of Yucatan, Mexico. Prev. Vet. Med. 57, 199–208. https://doi.org/10.1016/
S0167-5877(02)00230-1.
Snowder, G.D., Van Vleck, L.D., Cundiff, L.V., Bennett, G.L., 2006. Bovine respiratory
disease in feedlot cattle: environmental, genetic, and economic factors. J. Anim. Sci.
84, 1999–2008. https://doi.org/10.2527/jas.2006-046.
Straub, O.C., 1990. Infectious bovine rhinotracheitis virus. In: Dinter, Z., Morein, B.
(Eds.), Virus Infections of Ruminants. Oxford: Elsevier Science Publishers BV, pp.
71–108.
Takiuchi, E., Medici, K., Alfieri, A., Alfieri, A., 2005. Bovine herpesvirus type 1 abortions
detected by a semi-nested PCR in Brazilian cattle herds. Res. Vet. Sci. 79 (85), 88.
https://doi.org/10.1016/j.rvsc.2004.11.005.
van Schaik, G., 2001. Risk and economics of disease introduction to dairy farms. Tijdschr.
126, 414–418. (in Dutch). https://www.ncbi.nlm.nih.gov/pubmed/11436606.
van Schaik, G., Schukken, Y.H., Nielen, M., Dijkhuizen, A.A., Barkema, H.W., Benedictus,
G., 2002. Probability of and risk factors for introduction of infectious diseases into
Dutch SPF dairy farms: a cohort study. Prev. Vet. Med. 54, 279–289. https://doi.org/
10.1016/S0167-5877(02)00004-1.
van Schaik, G., Dijkhuizen, A.A., Huirne, R.B.M., Schukken, Y.H., Nielen, M., Hage, H.J.,
1998. Risk factors for existence of bovine herpes virus 1 antibodies on nonvaccinating
Dutch dairy farms. Prev. Vet. Med. 34, 125–136. https://doi.org/10.1016/S0167-
5877(97)00085-8.
Vonk-Noordegraaf, A., Labrovic, A., Frankena, K., Pfeiffer, D.U., Nielen, M., 2004.
Simulated hazards of losing infection-free status in a Dutch BHV1 model. Prev. Vet.
Med. 62, 51–58. https://doi.org/10.1016/j.prevetmed.2003.09.001.
Yesilbag, K., Bilge-dagalap, S., Okur-gumusova, S., Gunged, B., 2003. Studies on her-
pesvirus infections of goats in Turkey: prevalence of antibodies to bovine herpesvirus
1. Revue de Médecine Vétérinaire 154, 772–774. https://www.revmedvet.com/
2003/RMV154_772_774.pdf.