Comparative Immunology, Microbiology and Infectious Diseases 74 (2021) 101578

Contents lists available at ScienceDirect

Comparative Immunology, Microbiology and

Infectious Diseases
journal homepage: www.elsevier.com/locate/cimid

Current status and risk factors of canine parvovirus type 2 in North

Central Nigeria
Kenneth Ikejiofor Ogbu a, b, *, Ijeoma Chekwube Chukwudi b, Francesco Mira c,
Ukamaka Uchenna Eze b, Santina Di Bella c, Olushola Samuel Olaolu d,
Matthew Terzungwe Tion e, Giuseppa Purpari c, Vincenza Cannella c, Ignatius Chika Nwosuh f,
Annalisa Guercio c, Boniface Maduka Anene b
a
Department of Animal Health, Federal College of Animal Health and Production Technology, National Veterinary Research Institute Vom, Plateau State, Nigeria
b
Department of Veterinary Medicine, Faculty of Veterinary Medicine, University of Nigeria Nsukka, Enugu State, Nigeria
c
Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Via Gino Marinuzzi 3, 90129 Palermo, Italy
d
Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Ahmadu Bello University Zaria, Kaduna State, Nigeria
e
Department of Veterinary Medicine, College of Veterinary Medicine, Federal University of Agriculture Makurdi, Benue State, Nigeria
f
Viral Research Division, National Veterinary Research Institute Vom, Plateau State, Nigeria

A R T I C L E I N F O A B S T R A C T

Keywords: Since its emergence in Nigeria, canine parvovirus type 2 (CPV-2) infection has posed problems to dog breeding
Canine parvovirus and requires constant awareness and monitoring. In this study, the status, the assessment of extrinsic risk factors
Carnivore protoparvovirus 1 of parvoviral infection in dog kennels in North Central Nigeria, and isolation of the CPV-2 were carried out.
Risk factors
Potential risk factors were considered during sampling: age, breed, sex, location, vaccination and health status,
Prevalence
Vaccine
using well-structured questionnaires on dog owners with experience of CPV-2 infection. There was high prev­
Nigeria alence which depended on age, breed, location, clinical status of the dog while vaccination status of the dogs did
not influence the prevalence. CPV-2 vaccination compliance by the breeders and management system of the
kennels were also observed as risk factors. Isolation of CPV-2a and -2c strains from Nigeria for further study has
been reported. The spread of CPV-2 in Nigeria is increasing, hence needs for continual epidemiological moni­
toring and review.

1. Introduction
Canine parvovirus (CPV) is a member of the genus Protoparvovirus
(family Parvoviridae, subfamily Parvovirinae), included with other
related parvoviruses in the unique viral specie Carnivore protoparvovirus
1 [1]. The original canine parvovirus type 2 (CPV-2) emerged in the late
1970s [2] and was rapidly replaced by three CPV-2 variants (CPV-2a,
CPV-2b and CPV-2c) [2,3], worldwide widespread [4].
CPV-2 is a highly infectious virus causing acute and often fatal
gastroenteritis, affecting domestic and wild carnivores, transmitted by
direct or indirect contact through fecal-oral route [5]. Indeed, high
amounts of the virus particles are shed, mainly in faeces, for 7–14 days
or more after the onset of clinical signs [6]. Thus, clinical cases can be
due by direct contact with infected animals or indirectly by infected
fomites or passively transported by shoes, clothing, feeding bowls or
other animals. Although the use of effective and safe vaccines, CPV-2
have been reported to the leading cause of mortality and morbidity in
dogs [5,7].
Risk factors were associated with the development of clinical disease,
including stressors (such as early weaning, overcrowding and parasite
load), insufficient passive or active immunity, geographical region, and
the presence of other pathogens [8]. The disease has been reported to be
more severe in puppies than in adult dogs [9], although have been re­
ported that the infection can occur in all gender, age and breeds of dogs

* Corresponding author at: Department of Animal Health, Federal College of Animal Health and Production Technology, National Veterinary Research Institute
Vom, Plateau State, Nigeria.
E-mail addresses: drken2016@gmail.com (K.I. Ogbu), ijeoma.adieme@unn.edu.ng (I.C. Chukwudi), dottoremira@gmail.com (F. Mira), ukamakauchenna.eze@
unn.edu.ng (U.U. Eze), santinadibella78@gmail.com (S. Di Bella), olaoluolushola@gmail.com (O.S. Olaolu), tions_doc@yahoo.co.uk (M.T. Tion), giuseppa.
purpari@izssicilia.it (G. Purpari), vincenza.cannella@izssicilia.it (V. Cannella), chikanwosuh@gmail.com (I.C. Nwosuh), annalisa.guercio@izssicilia.it
(A. Guercio), boniface.anene@unn.edu.ng (B.M. Anene).

https://doi.org/10.1016/j.cimid.2020.101578
Received 18 August 2020; Received in revised form 22 October 2020; Accepted 3 November 2020
Available online 15 November 2020
0147-9571/© 2020 Elsevier Ltd. All rights reserved.
K.I. Ogbu et al.
[10]. The mortality rate can be higher (up to 70%) in puppies and
usually less than 1% in adult dogs [11]. Survival rates have been re­
ported to be as high as 80–95% when cases are symptomatically early
treated, but as low as 9.1% without treatment [12].
Since the emergence of CPV-2 in Nigeria in 1984 [13], it has posed
serious problems to dog breeding. The disease is usually prevalent in
unvaccinated dogs due to the limits in the owners’ compliance, high
costs of vaccines, poor husbandry, and faulty biosecurity practices [14].
The continuous presence of this pathogen, therefore, makes the disease
endemic in particular areas. The recent molecular analyses in Nigeria,
based on the partial VP2 gene [15–17] and the nearly full-length
genome sequences of CPV-2 [18], showed the circulation of all the
three CPV-2 variants (CPV-2a/2b/2c), and the spreading of further
genetically distinct strains in the last years [18]. These data arise new
questions about their impact in the local canine population and the
effectiveness of current use vaccines that need further investigations.
Laboratory diagnosis is usually based on the detection of CPV-2 in
the feces by hemagglutination test, electron microscopy, virus isolation,
direct immunofluorescence (IFD), PCR-based methods and by immu­
nochromatographic assays, which can also be used for rapid laboratory
confirmation of clinical diagnosis [6,7]. Concerns have been expressed
on the impact of the continuous evolution of CPV-2 to potentially
negatively affect the performance of diagnostic tests based on mono­
clonal antibodies and on the PCR-based assays [3]. Thus, continuous
reviews on the diagnostic tests should be assessed to limit the risks of
potential false-negative results and, then, to avoid the risk to favors the
CPV-2 spread.
During years, the evidence of mutations and genetic divergences of
CPV-2 suggested the need of further controls, as new strains of the virus
have continued to emerge in several parts of the world, including Africa
[16]. There are few and controversial reports about the risk factors
associated with CPV-2 infection in Nigeria [19,20]. The identification of
risk factors associated with canine parvovirus can also be used as
prognostic indicators for veterinarians and pet owners faced with CPV-2
infection [21] and to study new eradication plans of parvoviral enteritis.
In Nigeria, continuous monitoring of the CPV-2 infections is imperative
to for epidemiological review and determination of associated risk fac­
tors, which may be dependent on the location, hence this study.
2. Material and methods
2.1. Study area and population
This study was carried out in North Central Nigeria, one of the six
geo-political zones in Nigeria. This zone comprises six States (Plateau,
Nasarawa, Benue, Kogi, Niger and Kwara States) and Federal Capital
Territory (Abuja) namely. The study population comprised domestic
dogs presented to major Veterinary Clinics and dogs kept in major
kennels/breeder houses in the study area.
2.2. Sample size determination
The required sample size was determined using the formula as stated
by Thrusfield [22]:
Z 2 pq
n =
d2
(n = Desired sample size; Z = 1.96 (constant); p = Prevalence rate
from the average of previous studies; d = Desired absolute precision of ±
5% with 95% Confidence Interval; q = 1- p).
In this study, a prevalence rate of 17.14% according to Ogbu et al.
[23], was used for sample size determination. Using the above formula,
the calculated minimum sample size was 218, however an addition of
46.8% of the minimum sample size was done given a total sum of 320
dogs that were sampled in this study.
2
Comparative Immunology, Microbiology and Infectious Diseases 74 (2021) 101578
2.3. Sampling criteria
The study adopted descriptive survey design. Simple random sam­
pling method was used to select three States from the study area and
Federal Capital Territory (Abuja). These States and their capital cities
were: Plateau State (Jos; geographical coordinates: 9◦ 55′ 3.045′′ N,
8◦ 53′ 52.584′′ E; altitude: 1184 m a.s.l), Benue State (Makurdi;
7◦ 43′ 52.548′′ N, 8◦ 32′ 18.33′′ E; 90 m a.s.l), Nasarawa State (Lafia:
11◦ 21′ 23.178′′ N, 9◦ 23′ 8.202′′ E; 455 m a.s.l) and the Federal Capital
Territory (Abuja; 9◦ 3′ 51.59′′ N, 7◦ 29′ 21.47′′ E; 482 m a.s.l). Eight major
veterinary clinics, two from each State, and major kennels/breeders
were selected by purposive sampling method. Samples were collected
during the period from January to June 2018 from dogs of different
breeds with clinical signs of gastroenteritis and suspected of CPV-2
infection or apparently healthy. Scheduled visits were done to the pur­
posively selected veterinary clinics and kennels throughout the period of
study.
Potential risk factors were considered and registered during sam­
pling: age, breed, sex, location, vaccination status and health status. The
age of the animals was categorized into two classes: (A) 0–6 months and
(B) 6 months and above.
2.4. Sample collections
Rectal swabs were collected, and samples were preliminary in-clinic
screened by using an immunochromatographic test kit for detection of
CPV-2 antigen (SensPERT® Canine Parvovirus Test Kit, VetAll Labora­
tories, Korea), according to the manufacturer’s instructions. This chro­
matographic immunoassay for the qualitative detection of parvovirus
antigen in canine stool samples, as stated by the manufacturer’s in­
structions, was validated to detect the CPV-2a and CPV-2b variants and,
recently, the ability to detect also the CPV-2c variant was proved [18].
Positive samples were transported under controlled cold tempera­
ture to the Biotechnology laboratory, National Veterinary Research
Institute, Vom and then stored at − 20 ◦ C until they were further
analysed.
2.5. Virus isolation in cell cultures
In order to have viral isolates of the CPV-2 strain circulating in
Nigeria for future studies, CPV-2 positive rectal swab samples (n = 59)
were selected (based on age, sex, breed, vaccination status, clinical
status of the dogs and months of sampling). Samples were then sent to
the Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri” (Italy)
for molecular analyses, previously described [18], and viral isolation in
cell cultures. Those with higher amounts of CPV-2 DNA and selected
according to the CPV-2 variant and the area of collection, were pro­
cessed for the virus isolation and inoculated on permissive cell lines
A-72 (fibroblast-like cells derived from canine tumour) [24].
A-72 cells were maintained in culture medium (Eagle’s Minimum
Essential Medium (EMEM), Sigma–Aldrich®, Milan, Italy) supple­
mented with 10% Fetal Bovine Serum (EuroClone®, Milan, Italy) and an
antibiotic and antimycotic solution (100 U/mL penicillin G sodium salt,
0.1 mg/mL streptomycin sulfate, 0.25 μg/mL amphotericin B; Euro­
Clone®, Milan, Italy). Rectal swabs were then inoculated on cell
monolayers in the mitotic phase, left in contact for 30 min at 37 ◦ C and
5% CO2. Culture medium was added, and the incubation was performed
at 37 ◦ C and 5% CO2 for 6–7 days. Inoculated cells were monitored daily
and, according to the laboratory procedures, two additional blind pas­
sages were carried out before considering virus isolation as unsuccessful.
Viral growth was evaluated by detection of cytopathic effect (CPE) and
PCR. Thus, monolayers were subjected to three cycles of freezing-
thawing, centrifuged at low speed (1.500 x g for 10 min at 4 ◦ C) and
the collected supernatant was tested for parvovirus DNA by PCR assays
and immunofluorescence, to confirm the presence of the infectious
virus. The nearly complete genome sequence of the CPV-2 strains was
K.I. Ogbu et al. Comparative Immunology, Microbiology and Infectious Diseases 74 (2021) 101578

sequenced, as previously described [18]. Table 1

For the direct immunofluorescence assay, each sample was firstly Prevalence of CPV-2 in naturally infected dogs based on age, sex, breed, loca­
inoculated on a glass chamber slide with A72 cells in mitotic phase using tion, vaccination and health status distributions in North Central Nigeria.
the EMEM medium. Incubation was performed at 37 ◦ C and 5% CO2, Parameter Positive Negative Total Prevalence (%)
read every day, for 1–4 days. Secondly, the medium was eliminated, the Puppy 121 125 246 49.19
chamber was moved, and the slide was dried. The slide was fixed with Adult 23 51 74 31.08
acetone at − 20 ◦ C for at least 30 min and then stained using specific Total 144 176 320 45.00
monoclonal antibodies against CPV-2 (VMRD, Inc, Pullman, WA, USA) p – Value = 0.007; df = 1; Chi square value = 7.535
in accordance with the manufacturer’s instructions.
Male 79 108 187 42.25
Female 65 68 133 48.87
Total 144 176 320 45.00
2.6. Determination of risk factors of CPV-2 infection in dog kennels p – Value = 0.256; df = 1; Chi square value = 1.379

To further define the risk factors associated with CPV-2 infection in Caucasian 33 19 52 63.46
Nigeria, questionnaires were administered to dog owners whose dogs Rottweiler 29 12 41 70.73
German shepherd 12 13 25 48.00
have had diagnosed the CPV-2 infection. The study population was Local breed 15 20 35 42.86
determined using focal point census of the study area while the sample Mixed breed 12 28 40 30.00
size was determined using a percentage of the latest census population of Lhasa apso 6 8 14 42.86
dogs in the study area. The total number of dogs in Jos metropolis is Bull mastiff 7 14 21 33.33
Boer bull 10 26 36 27.78
estimated to be about 10,024 [25]. It was estimated that the ratio of dog
St. bernard 6 11 17 35.29
to owner is 4:1 which will give approximately 2506 dog owners in the Neapolitan mastiff 11 18 29 37.93
selected areas in North Central Nigeria. Using approximately 20% of this Chihuahua 3 7 10 30.00
owners’ population, 500 well-constructed questionnaires were admin­ Total 144 176 320 45.00
istered to dog owners as the sample size, according to Boll and Gall p – Value = 0.001; df = 10; Chi square value = 29.558

model cited in Uzoagulu [26].These questionnaires had the aim to

Plateau State 54 26 80 67.50
collate data on the following: CPV-2 vaccination compliance among dog Nasarawa State 23 57 80 28.75
breeders in North Central Nigeria, management system in dog breeders’ Benue State 35 45 80 43.75
kennels, and biosecurity measures in dog breeders’ kennels in North Fct Abuja 32 48 80 40.00
Central Nigeria. Total 144 176 320 45.00
p – Value = 0.000; df = 3; Chi square value = 25.758

Vaccinated 69 95 164 42.07

2.7. Statistical analysis Not vaccinated 75 81 156 48.08
Total 144 176 320 45.00
The data obtained were analysed using descriptive statistics and Chi- p – Value = 0.3122; df = 1; Chi square value = 1.164
square test. Significance was accepted at probability values of p < 0.05.
Clinical suspect 119 123 242 49.17
Results were presented in graph and charts.
Apparently healthy 25 53 78 32.05
Total 144 176 320 45.00
3. Results p – Value = 0.009; df = 1; Chi square value = 6.987

3.1. Prevalence of CPV-2 in naturally infected dogs in North Central

3.1.4. Geographical distribution
Nigeria
Location had a strong association with the prevalence of CPV-2
infection (p = 0.000). Based on location, the prevalence of CPV-2 was
The result showed that the prevalence of CPV-2 in the study area was
highest in Plateau state recording 67.50% (54/80) while Nasarawa had
45% (144/320). According to the potential risk factors considered and
the least prevalence of 28.75% (23/80).
registered during sampling, different prevalence and significances were
observed and reported in Table 1:
3.1.5. Vaccination status distribution
The prevalence of CPV-2 among vaccinated dogs was 42.07% (69/
3.1.1. Age distribution
164) while the prevalence among unvaccinated dogs was 48.08% (75/
The prevalence of CPV-2 among puppies (0–6 months) was 49.19%
156). There was no strong association (p = 0.3122) between the pres­
(121/246) while the prevalence among adults was 31.08% (23/74).
ence of CPV-2 and vaccination status distribution.
There was a strong association (p = 0.007) between the presence of CPV-
2 and age distribution.
3.1.6. Health status distribution
There was a strong association (p < 0.009) between the prevalence of
3.1.2. Sex distribution
CPV-2 and clinical manifestation distribution. The clinical signs
The prevalence of CPV-2 among males was 42.25% (79/187) while
observed among the dogs suspected to be infected with CPV-2 were
the prevalence among females was 48.87% (65/133). There was no
vomiting, foul smelling diarrhoea (with and without blood), emaciation,
strong association (p = 0.256) between the presence of CPV-2 and sex
lethargy, fever, tachycardia, and sunken eyes.
distribution.
3.1.7. Case mortality
3.1.3. Breed distribution
The case mortality among those infected with CPV-2 infection (n =
The prevalence of CPV-2 based on breed showed that Rottweiler had
144) was 80.6% while the survival rate was 19.4%.
the highest prevalence of 70.73% (29/41) while Boer bull had the least
prevalence of 27.78% (10/36). The prevalence among the indigenous
breed (local breed) was 42.86% (15/35) while that of mixed breed was 3.2. Virus isolation
30.00% (12/40). There was a strong association (p = 0.001) between the
presence of CPV-2 and breed distribution. From the total CPV-2 positive samples, n = 12 samples were selected,

3
K.I. Ogbu et al. Comparative Immunology, Microbiology and Infectious Diseases 74 (2021) 101578

based on the quantity of CPV-2 DNA and on the area of origin, and were
inoculated into cell lines. All these samples were isolated from A-72 cell
monolayers after multiple passages and the isolation was confirmed by
the immunofluorescence and PCR assays (Supplementary Material:
Supplementary Table 1 and Supplementary Figures 1 and 2). CPV-2a
strains were isolated at the 3rd blind passage while CPV-2c strains
were isolated at the 2nd passage. Isolates of CPV-2a (n = 1) and CPV-2c
(n = 8) from this study were sent to be preserved and long term stored at
the Biobanca del Mediterraneo (www.bbmed.it), a biobank for the safe
storage of biological material. The nearly complete genome sequences of
these CPV-2 strains were also obtained, as previously described [18].
These sequence data have been submitted to the DDBJ/EMBL/GenBank
databases under accession numbers MT840286-94.
3.3. Risk factors of CPV-2 infection in dog kennels in North Central
Nigeria
3.3.1. CPV-2 vaccination compliance of dog breeders
Out of the total number of respondents (n = 500), 41.6% (208/500)
agreed that they vaccinate their dogs against CPV-2 although, 52.4%
(109/208) had their dogs administered only primary vaccination while
47.6% (99/208) gave both primary and secondary vaccinations
(Table 2). There was an association between CPV-2 infection in dog
kennels and its vaccination compliance by the dog owners (p = 0.024).
3.3.2. Management system in dog breeders’ kennel
Majority of the respondents (58.4%) feed puppy and adult dogs
together while 41.6% (208/500) feed puppies separate from adult. Out
of the total number of respondents that feed the dogs separately, 56.2%
(117/208) feed the puppy before adult. Majority of the respondents
(79.2%) stated that they feed puppy and adult dogs with the same
trough (feeder) (Table 2). It was also shown that there was an associa­
tion between feeding pattern of dog breeders and CPV-2 infection in the
dog kennels in the study area (p = 0.00).
Majority of dog owners use cemented floor kennels (58.4%) while
few use wooden floor kennel (19%) (Table 2). The cleaning rate of the
kennel vary as 41.4% of the respondents clean their kennel daily while
9.8% clean on weekly basis. Majority of the respondents (48.8%) do not
have regular rate of cleaning. Furthermore, most of the owners (58.4%)
do not use detergent to clean the dog’s kennel (Table 2). The results
showed that the kennel’s floor pattern and cleaning rate was signifi­
cantly associated with CPV-2 infection in the dog kennels in North
Central Nigeria (p = 0.00).
3.3.3. Biosecurity measures
Although most of the respondents (63%) do not allow visitors to their
kennel, only 43% wash their hand before attending to their dogs. Out of
the total respondents, 54.8% quarantine their new dogs while only few
breeders (45.2%) isolate their dogs in disease conditions (Table 2).
Based on the result from the study, there was an association between the
CPV-2 infection in the dog kennels and the biosecurity measures prac­
ticed by the dog owners (p = 0.00).
Out of the total respondents, 64.6% confine their dogs while 35.4%
allow their dog to scavenge. Among those that confine their dogs, ma­
jority of them confine dogs of different litters (62.2%), different ages
(52%) and different breeds (56.7%) together (Table 2). The results also,
showed that CPV-2 infection in the dog kennels is significantly associ­
ated with confinement of the dogs by the owners in the study area. (p =
0.04).
4. Discussion
Canine parvovirus (CPV-2) has a worldwide distribution and, since
its emergence in the late 1970s, continues to spread in the domestic and
wild carnivore populations [4]. Since 1984 was described in Nigeria
[13] and has continued to circulate causing often fatal gastroenteritis in
dogs leading to high mortality and morbidity rates [18]. Despite the
availability of commercial vaccines in Nigeria, CPV-2 is a threat and still
causes great losses in the local canine population [5].
Based on this study, the overall prevalence of CPV-2 in North Central
Nigeria was 45%. This result suggests a continuous circulation of the
CPV-2 in the canine population, where almost the half of the tested dogs
showing clinical signs of gastro-enteric disease were referable to this
viral infection. Indeed, despite other potential viral etiological agents

Table 2
CPV-2 vaccination compliance of dog breeders, management system and biosecurity measures in dog breeders’ kennel in North Central Nigeria.
Vaccination compliance Level of compliance

Yes No Total Primary Secondary Total

208 292 500 109 99 208
41.6% 58.4% 100% 52.4% 47.6% 100%
P = 0.024; df = 2; Chi square value = 7.433

Feed puppies and adults together Feed puppies before adults Separate trough for puppies/adults

Yes No Yes No Yes No

292 208 117 91 104 396
58.4% 41.6% 56.2% 43.8% 20.8% 79.2%
P = 0.000; df = 2; Chi square value = 164.197

Floor types Cleaning rate Use of detergents

Wooden Cemented Wire Daily Weekly Others Yes No

195 192 113 207 49 244 208 292
39% 38.4% 22.6% 41.4% 9.8% 48.8% 41.6% 41.6%
P = 0.000; df = 6; Chi square value = 241.057

Allowing of visitors Washing of hands Quarantine new dogs Isolation of sick dogs

Yes No Yes No Yes No Yes No

185 315 215 285 274 226 274 226
37% 63% 43% 57% 54.8% 45.2% 45.2% 54.8%
P = 0.000; df = 3; Chi square value = 47.537

Confinement Confine different litters Confine different ages Confine different breeds

Yes No Yes No Yes No Yes No

323 177 201 122 171 152 183 140
64.6% 35.4% 62.2% 37.8% 52.9% 47.1% 56.7% 43.3%
P = 0.004; df = 3; Chi square value = 13.230.

4
K.I. Ogbu et al.
were tested [18], CPV-2 was the main viral agent of gastroenteritis in
dogs, as observed by other authors [27]. The continuous changes in the
global scenario, due to the intrinsic high rates of mutation of the CPV-2
[17,28] together with the high resistance in the environment [29], could
have also contributed to these high rates. Moreover, the changes
observed in the most recent CPV-2 strains spreading in Nigeria [18]
potentially due to a recent introduction from other Countries or conti­
nents connected with the trading and transport of dogs [30–32], could
have also contributed to these high rates.
Compared to other previous studies in Nigeria, this prevalence is
higher than 13.4% [19], 17.14% [23] and 37.7% [33] values, and
slightly lower than prevalence of 47.7% and 75% recorded by Chollom
et al. [5] and Apaa et al. [16], respectively. The lower prevalence
recorded in this research compared to some author’s reports [5,16], may
be due to the different statistical sample examined and to PCR used in
their diagnosis which is a higher sensitive diagnostic technique [34]
compared to immunochromatographic assay used in this study, whose
sensitivity is slightly lower [35,36].
Although CPV-2 have been known to be ubiquitous and gained
worldwide occurrence [4], the prevalence of the virus in this study area
was shown to significantly vary also within the North Central Nigeria,
where Plateau State had the highest prevalence. Indiscriminate impor­
tation of dog with its attendant introduction of diseases from other
countries into Jos Plateau State, which appears to be the hub of dog
breeding in Nigeria, could lead to changes in the current spreading
CPV-2 strains [18]. Moreover, different climatic features could have
fostered the local viral spreading: indeed, the prolonged cold weather of
Jos, Plateau and the similar climate/weather conditions in Benue State
and Federal Capital Territory could be related to the seasonality of the
CPV-2 infection, as observed by Shima et al. [37]. This study is the first
report on CPV-2 infection in Nasarawa State of Nigeria where the lowest
prevalence was recorded, and this may be due to the lack of interest in
pet keeping by most of the inhabitants of the State. Further evaluations
are necessary to better assess the specific connection of the weather and
socio-economical variants to elucidate their potential inference in the
local viral spreading.
Due to the virus intrinsic characteristics, early and time-consuming
diagnosis of CPV-2 infection is necessary to allow the early isolation of
infected dogs and prevention of the spread of the disease to the sus­
ceptible animals. To achieve a conclusive clinical diagnosis sensitive and
specific laboratory techniques have been developed [36]. Most of these
techniques require specialized laboratories and personnel to be carried
out, which resources are often limited. Quite the opposite, immuno­
chromatography (IC) is the most common, sensitive, specific, and rapid
field diagnostic technique used as the test procedure is easy, simple, and
rapid [34,38]. Concerns have been expressed regarding the ability of
these in-clinic tests to detect accurately the CPV-2c variant with the
same accuracy as other variants [39]. Further studies have solved this
concerns demonstrating the ability of an in-clinic assay to detected
easily also the CPV-2c variants [35] also despite the account of further
mutations due to its continuative adaption and evolution [18]. There­
fore, this study further corroborates the effective used to detect the
presence of all variants CPV-2 antigens in fecal swab samples of dogs.
The study revealed that the prevalence of CPV-2 is related to the age
of the dog as the prevalence was significantly higher in puppies than in
adult dogs, in agreement with previous studies [40] also conducted in
the same Country [19,23,33]. The higher prevalence in puppies could be
due to their maternally derived antibodies (MDA) interference which
may hinder conferment of protective immunity post-vaccination and
results too low to protect puppies during active infection [11] and/or to
the high affinity to rapidly dividing cells [41], and may explain this
higher prevalence in puppies. It could also be a result of
immuno-incompetence of the puppies’ immune system or lack of
adequate maternal antibodies in their early stage of life [42]. In addi­
tion, Ukweze et al. [33] reported that adult dogs may be less susceptible
to CPV-2 infection probably due to age-reduced susceptibility,
5
Comparative Immunology, Microbiology and Infectious Diseases 74 (2021) 101578
vaccination-induced specific immunity, or previous and often subclini­
cal infection-induced specific immunity.
Although it was previously reported that vaccination is the most
effective method of controlling CPV-2 infection [29], the result of this
study revealed that prevalence of CPV-2 infection was not dependent on
the vaccination status of the dogs. It is due to the evident
non-significance difference in the prevalence of CPV-2 between vacci­
nated and unvaccinated dogs, and a high rate (42.07%) of positive dogs
were vaccinated. The result disagreed with many authors who reported
significant difference between vaccinated and unvaccinated dogs in the
prevalence of CPV-2 infection [29,37,43]. This may be due, as
mentioned above, to the presence of MDA in the puppies but also to the
improper vaccine and vaccination managements, low or non-responders
individuals to vaccinations, as corroborated by Spibey et al. [44] and
Larson & Schultz [43]. However, recent studies by Chiang et al. [45] and
Hoang et al. [46] reported that 22 and 44, respectively, of
CPV-2c-diseased dogs from Taiwan and Vietnam were confirmed to be
vaccinated against CPV-2 and four vaccinated dogs from Taiwan,
including an adult dog with completed vaccination protocol, died
despite vaccination [45]. The CPV-2c strains recently spreading in
Nigeria [18] are genetically related to the CPV-2c strains from Taiwan
and Vietnam, and some of the observed amino acid changes also in these
studies may cause changes in the tertiary structure of the protein [47],
potentially weakening the binding of specific antibodies and, therefore,
creating concerns about the virus ability to evade the immune system.
Therefore, a re-evaluation of the efficacy of CPV-2 vaccines currently
used against these most recent CPV-2c strains [45,46] become a recent
future challenge.
The prevalence of CPV-2 infection was shown to be influenced by the
breed of dogs, in consonance with other studies in Nigeria [19,33]. The
result of this study recorded a breed susceptibility pattern, with the
highest in Rottweiler, Caucasian, German shepherd (Alsatian) and local
(indigenous) breeds, as also observed by other Authors [5,10,21,33,48,
49]. The higher prevalence of CPV-2 infection in the indigenous breed
than some exotic breeds disagreed with Nelson & Couto [50], who
opined that local breed of dogs are less susceptible to CPV-2 infection
than exotic breeds.
The prevalence data of this study suggests also that in this Country is
necessary to introduce routine serological tests before and/or after CPV-
2 vaccination to assess the residual level of MDA or the seroconversion
of vaccinated dogs, respectively. In particular, is needed for those pure
or local breeds considered as potential low-responders and non-
responders [51], as suggested by the guideline of the Vaccination
Guidelines Group (VGG) of the World Small Animal Veterinary Associ­
ation (WSAVA) [52], specially before to consider the vaccination as
non-protective. Moreover, attentions should be also focused on the
vaccine management, such as correct storage or transportation and
handling of the vaccines, to reduce the risks of inactivation of the MLV
products and, then, of their potential ineffectiveness [52].
Based on the breeders (respondents) reports, the main risk factors
associated with the occurrence of the CPV-2 infection in the kennels
were the vaccination compliance by the breeders and the management
system of the kennel, such as feeding pattern, floor pattern, rates of
cleaning the kennel and biosecurity measures adopted by the breeders in
their kennels. Despite vaccination is the most effective method to control
the infection [29], the non-compliance to the vaccination may be due to
unawareness of some breeder, the high costs of the vaccines and esti­
mated value of the dogs [19]. In addition, the control of the CPV-2
infection is dependent on cleanliness of the kennel [53] together with
biosecurity measure and good hygiene [5,19,37]. For these purposes,
effective and low costs disinfectants such as sodium hypochlorite and
correct disinfection protocols [54] could be suggested to the breeders to
control and limit the CPV-2 spreading.
K.I. Ogbu et al.
5. Conclusion
There is high prevalence of CPV-2 infection in the study area which
dependent on age, breed, location, clinical status of the dog while
vaccination status of the dogs seemed not have influenced the preva­
lence of the infection. Immunochromatographic test can be used for
rapid detection of CPV-2 in fecal samples of dogs. CPV-2 vaccination
compliance by the breeders and management system of the kennel are
the main risk factors associated with previous events of CPV-2 infection.
Therefore, further studies are suggested to evaluate the implication of a
novel CPV-2 mutant on the effectiveness of current use vaccines and to
evaluate the production of new vaccines based on the circulating CPV-2
strains. Adequate vaccination and proper hygiene in the management of
dogs rearing by dog owners, adherence to World Small Animal Veteri­
nary Association (WSAVA) vaccination guidelines by veterinarians and
the new laws on importation of dogs into Nigeria also to control the
importation of infectious agents into the country should be
recommended.
Funding
This work was supported by the Ministero della Salute (Italy) [grant
number Ricerca Corrente IZS SI 03/18 RC “Studio del potenziale zoo­
nosico e caratterizzazione genomica dei virus enterici del cane”].
Role of the funding source
The funding source had no involvement in study design; in the
collection, analysis and interpretation of data; in the writing of the
report; and in the decision to submit the article for publication.
Declaration of Competing Interest
None.
Acknowledgements
The authors wish to acknowledge the following for their contribu­
tions: Dr. Gabriele Chiaramonte (Istituto Zooprofilattico Sperimentale
della Sicilia “A. Mirri”, Palermo, Italy) and Prof. Garba Hamidu Shar­
ubutu (Agricultural Research Council of Nigeria, Abuja, Nigeria), Vet­
erinary clinics and dog breeders.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.cimid.2020.101578.
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