https://doi.org/10.52973/rcfcv-e33221
Received: 27/12/2022 Accepted: 27/02/2023 Published: 03/03/2023
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Revista Científica, FCV-LUZ / Vol. XXXIII, rcfcv-e33221, 1 – 7
ABSTRACT
The aim of this study was to identify the presence of tick-borne
pathogens as the probable cause of reproductive problems in cows in
a commercial dual-purpose cow operation living in tropical conditions
in Northern Veracruz, Mexico. As part of the reproductive control
of the herd, ultrasonography was performed periodically, and the
presence of three open (non-pregnant) cows with historical infertility
problems was detected. The ovaries from the two open cows and
the mummied foetus of a third one were removed, and two sets
of tissue samples were collected. Histopathology analysis revealed
the presence of follicular abnormalities. The DNA tests revealed the
presence of Anaplasma marginale, Babesia bigemina and/or Babesia
bovis. In conclusion, the present work represents the rst report of
tick-borne pathogens in ovarian tissue samples and a mummied
foetus of infertile cows in tropical conditions in Northern Veracruz,
Mexico, highlighting the possibility that many reproductive problems
in tropical conditions could be caused by tick-borne pathogens.
Key words: Abortion; bovine anaplasmosis; bovine babesiosis;
infertility; tropical reproductive diseases
RESUMEN
El objetivo de este estudio fue identicar la presencia de patógenos
transmitidos por garrapatas como la causa probable de problemas
reproductivos en vacas procedentes de una explotación comercial
de doble propósito que viven en condiciones tropicales en el norte
de Veracruz, México. Como parte del control reproductivo del rebaño,
se realizó ecografía periódicamente y se detectó la presencia de
tres vacas abiertas (no preñadas) con historia de infertilidad. Se
extrajeron los ovarios de las dos vacas abiertas, así como el feto
momificado de una tercera, y se obtuvieron dos conjuntos de
muestras de tejido. El análisis histopatológico reveló la presencia
de anomalías foliculares. Las pruebas de ADN revelaron la presencia
de Anaplasma marginale, Babesia bigemina y/o Babesia bovis. En
conclusión, el presente trabajo representa el primer reporte de
patógenos transmitidos por garrapatas en muestras de tejido ovárico
y un feto momicado de vacas infértiles en condiciones tropicales
en el norte de Veracruz, México, destacando la posibilidad de que
muchos problemas reproductivos en condiciones tropicales puedan
ser causados por patógenos transmitidos por garrapatas.
Palabras clave: Abortos; anaplasmosis bovina; babesiosis bovina;
infertilidad; enfermedades reproductivas tropicales
Molecular detection of Tick-Borne Pathogens in Ovaries and a Mummied
Foetus of three cows with Historical Reproductive Problems in Northern
Veracruz, Mexico
Detección molecular de patógenos transmitidos por garrapatas en ovarios y fetos momicados de
tres vacas con historia de problemas reproductivos en el norte de Veracruz, México
Miguel Angel Lammoglia-Villagómez
1
, Amalia Cabrera-Núñez
1
, Rebeca Rojas-Ronquillo
1
, Jorge Luis Chagoya-Fuentes
1
, Ingeborg Becker
2
and Sokani Sánchez-Montes
1,2
*
1
Universidad Veracruzana, Facultad de Ciencias Biológicas y Agropecuarias región Tuxpan. Tuxpan de Rodríguez Cano, Veracruz, México.
2
Universidad Nacional Autónoma de México, Facultad de Medicina, Unidad de Medicina Experimental, Centro de Medicina Tropical. Ciudad de México, México.
*Corresponding author: sok10108@gmail.com
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INTRODUCTION
Reproductive diseases represent the main cause of economic
losses in dairy and beef cattle (Bos taurus) production [19]. Abortions
and stillbirths in productive animals are estimated to represent an
approximated cost between USD 500 and 900 per case [3, 11]. In
general, the main agents that can cause abortive syndromes include
viruses (bovine viral diarrhoea, bovine viral rhinotracheitis), bacteria
(Leptospira spp., Brucella spp.) and parasites (Toxoplasma gondii,
Neospora caninum) [12]. In tropical environments, several tick-borne
microorganisms have a signicant impact on the reproductive health
of production animals [5, 10]. On the American Continent, bovine
babesiosis (caused by Babesia bigemina and Babesia bovis) and
bovine anaplasmosis (caused by Anaplasma marginale), are the two
major diseases responsible for production losses of around 100,000
million USD [14]. In Mexico, both tick-borne diseases are endemic.
Various serological and molecular studies have shown a prevalence
higher than 50%, particularly in the States of Coahuila, Nuevo León,
Tamaulipas, and Veracruz [6, 17].
Both groups of pathogens produce fever, which has a negative
impact on pregnant animals since hyperthermia can affect early
embryo implantation and/or the establishment and development of the
placenta. Some historical studies have suggested that these organisms
do not have the ability to penetrate the placenta, although there is new
evidence that they can infect the foetal internal organs [9].
It is noteworthy that reports of abortions caused by anaplasmosis
and babesiosis in bovines are historical, scattered and scarce: A.
marginale was recorded in stillbirths of Brazil between 1974–1975,
whereas B. bovis was recorded in an abortion in Australia in 1986
[4, 20]. Experimental trials in South Africa have demonstrated that
animals born to females infected with A. marginale develop antibodies
weeks after birth and, if they were splenectomised, that they become
chronic carriers of the pathogen [16]. Serological studies have shown
that a high prevalence of A. marginale infection is correlated with a
high frequency of abortive events. Yet, the rst study in which A.
marginale and B. bovis were molecularly identied in abortions was
carried out in Brazil between 2018 and 2019 [9].
For Mexico, there are no studies where these tick-borne pathogens
have been correlated with mummied foetuses or ovaries from
infertile cows. Therefore, the aim of the present study was to identify
the presence of tick-borne pathogens in the ovaries from two infertile
cows and the internal organs from a mummied foetus of a third one
in a region with high prevalence of A. marginale and B. bovis in the
North of Veracruz, Mexico.
MATERIALS AND METHODS
This study was performed in a private dual-purpose cow Bos
taurus × Bos indicus operation in a tropical warm, high-humidity
region in northern Mexico (20°57'0"N, 97°24'0"W). The vaccination
program against diseases that affected reproduction consisted of:
a) Annual application (January) of inactivated virus vaccine against
Infectious Bovine Rhinotracheitis (IBR), Bovine Viral Diarrhoea (BVD)
biotype 2 – non-cytopathic, Parainuenza 3 (PI3), Bovine Respiratory
Syncytial Virus (RSBV) and bacterin against: Leptospira interrogans
serovars: canicola, grippotyphosa, icterohaemorrhagiae, pomona
and Leptospira borgpetersenii serovar hardjo-bovis and, b) Annual
application (July) of bacterin against: Leptospira interrogans serovars
canicola, grippotyphosa, icterohaemorrhagiae, pomona and Leptospira
borgpetersenii serovar hardjo-bovis. As part of the reproductive
control of the herd, ultrasonography was performed periodically
using a CONTEC Color Doppler Ultrasonic Diagnostic Device (CONTEC,
CMS1700A, Singapore). All the cows in the herd were evaluated 21
days after calving and did not present any pathologies. At 80 days
postpartum, a second reproductive diagnostic was carried out using
ultrasonography and again, the diagnosis was free of pathologies and
in optimal conditions to start the synchronization protocol Ovsynch.
All cows showed oestrus 21 days after being rst bred and they were
bred two more times. After one oestrous cycle following treatment, the
cows were synchronized using oestradiol benzoate and an intravaginal
device containing 1.9 grams (g) of progesterone. During this evaluation
the presence of three non-pregnant females with historical infertility
problems was detected. Of the three cows involved in this research,
two of them had a breed composition of 3/8 Bos indicus (Gyr) and 5/8
Bos taurus (Holstein) while the other cow had 3/8 Bos indicus (Gyr) and
5/8 Bos taurus (Brown Swiss).
Hormonal treatments were implemented considering that the
cause of the reproductive problem was an endocrine disorder, which
was corrected in one of them, becoming pregnant. The other two
cows remained open and were treated with antibiotics (intrauterine
infusion and intramuscularly), considering that the infertility could be of
infectious origin. A systemic treatment with antibiotic (oxytetracycline
LA, 200.0 miligrams· kilograms
-1
–mg·kg
-1
–) was carried out and repeated
at 72 hours. Additionally, an intrauterine infusion with 2,000 mg of
oxytetracycline in 50 mL of physiological saline solution was also
performed and was repeated at 72 hours. Fertility did not improve, for
which it was decided to implement a combined scheme of hormones
and antibiotics. It is important to mention that Amblyomma mixtum
and Rhipicephalus microplus ticks are endemic in this region, as well
as the diseases they can transmit (Babesiosis and Anaplasmosis).
However, the three cows never presented any signs of diseases caused
by these haemoparasites. Additionally, these two cows were placed in
an enclosure with a bull of the same race for a period of 65 days to try to
get them pregnant; however, at the end of the period, both specimens
remained open. After periodic ultrasonography, the development
of persistent follicular cysts was observed in one of them (FIG. 1A),
and the other cow had developed a corpus luteum (CL) with a very
echogenic but small area that had been growing for several weeks and
was diagnosed as a persistent CL (FIG. 1B). The third cow, the one that
became pregnant, was diagnosed with a mummied foetus.
The ovaries from the two non-pregnant cows were removed, and
tissue samples were collected.
In the third cow, the mummied foetus was recovered by c-section
(FIG. 1C and 1D), and a total of 30 samples of different organs (brain, lung,
cardiac muscle, skeletal muscle, among others) were collected (FIG. 2C.).
Two sets of tissue samples of the mummied foetus (including
the ovaries of the two open cows) were collected, one of them was
immersed in formaldehyde solution at 5% and sent to the diagnostic
laboratory for histopathological diagnose; the other sample set was
xed in 70% ethanol for Deoxyribonucleic acid (DNA) extraction.
Histopathological studies were carried out using haematoxylin-eosin
(H-E) staining to determine changes in sample tissues.
Genomic DNA was extracted individually from a portion of 25 mg
of tissue, using the QIAamp® DNA Mini Kit (QIAGEN, Hilden, Germany)
according to the manufacturer’s specications (using the Purication of
Total DNA from Animal Tissues Protocol). As an endogenous control of
the extraction, a fragment of ≈708 base pairs (bp) of the mitochondrial
A B
C D
FIGURE 1. Ovaries from adults (A, B) and fetuses (C, D) with anaplasmosis, babesiosis, or both conditions
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FIGURE 2. Histopathological results: (A) case one (left ovary): small
follicular cysts were observed in the ovary cortex (arrow). (B) case one
(right ovary) calcication process in the cyst wall (arrow). (C) case two
(right ovary) calcication in the cyst wall (arrow)
A
B
C
Tick-Borne pathogens in cows with reproductive problems / Lammoglia-Villagómez et al. _________________________________________
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gene Cytochrome Oxidase Subunit 1 (COI) was amplied, using the
primers LCO1490 and HCO2198 [7]. For Ehrlichia/Anaplasma detection,
it was used the primers EHR16SD/EHR16SR, which amplied a fragment
of 345 bp of the ribosomal gene 16S rDNA [15].
For Brucella DNA detection, conventional polymerase chain reaction
probe (PCR) was performed in a Veriti 96-Well Fast Thermal Cycler
(ThermoFisher Scientic, 4375305, Massachusetts,USA) using the
primers F4/R2, proposed by Romero et al. [18], which amplify a
900-bp fragment of the ribosomal 16S-rDNA gene (16S). In the case
of Leptospira detection, it was amplied a fragment of 400 bp of the
32-kilodalton (Kd) lipoprotein, using the primers Lip32+/Lip32 – [21].
Finally, Babesia/Theileria were detected using the primers BAB01/
BAB02 [2]. The reaction mixture was prepared in a nal volume of
25 microliters (µL) reaction mix, with 12.5 µL of GoTaq® Green Master
Mix, 2X Promega Corporation (Madison, WI, USA), 1 µL of each primer
(100 nanograms (ng) each), 10µl DNA (~50 ng) and 0.5 µL nuclease-
free water. It was included positive (DNA of Anaplasma ovis, Brucella
mellitensis, Ehrlichia canis, Leptospira borgpetersenii and Theileria
equi) and negative (ultrapure water instead of DNA) controls.
The amplication products were subjected to electrophoresis in
2% agarose gel [15] stained with Smartglow. The positive PCR product
was sequenced at Macrogen Inc., Korea.The sequence was compared
with those of the references deposited in GenBank, using the BLASTn
tool as a preliminary conrmation of the presence of bacterial and
parasitic sequences. The sequences obtained were deposited in
GenBank under accessions numbers MZ798902 for A. marginale (16S
rDNA) and MZ798903–MZ798904 for B. bigemina and B. bovis (18S
rDNA). This study was approved by the Bioethics Committee of the
Facultad de Ciencias Biológicas y Agopecuarias, Campus Tuxpan of
the Universidad Veracruzana (UV). Animals were handled according
to National Legislation and Ethics (NOM-012-ZOO-1993).
RESULTS AND DISCUSSIONS
In the case of the rst non-pregnant cow with a history of infertility,
the presence of a chronic follicular cyst in one of the ovaries was
detected by ultrasonography (FIG. 2A).
After surgical extraction, the presence of a follicular cyst with a thick
calcied wall and an accumulation of 14 millilitres (mL) of follicular uid
was conrmed (FIG. 2A), and the other ovary of this cow presented an
exaggerated number of growing follicles, which were diagnosed as
pathological (FIG. 2B). Histopathological analysis of the samples revealed
the following ndings: In the case of the non-pregnant cow diagnosed
with follicular cyst, an exaggerated population of growing follicles was
observed in one ovary, and the histopathological diagnosis was the
presence of follicular cysts in the ovary cortex (FIG. 2A) and in the ovary
that presented the chronic follicular cyst (FIG. 2B); a surface with a
degree of calcication was macroscopically identied, and the tissue
continued with a calcication that changed to a yellow colour (FIG. 2C).
Microscopic description indicated a benign cystic tumour with a
calcied wall, fragmented and lined by serous-type simple cuboidal
epithelium whose nuclei did not show atypia or mitoses and was
considered negative for neoplasms (FIG. 2 A-C).
The second non-pregnant cow and with a history of infertility had
developed a persistent CL with a highly echogenic area, diagnosed
by ultrasonography. After surgical removal, a CL with calcications
was identied. Subsequently, a CL was observed in the ovary that
had a calcied membrane, and histopathological analysis described
1 2 3 4 5 6 7
1 2 3 4 5 6 7 8
FIGURE 3. Agarose-gel electrophoresis of the PCR products amplied
with primers for A) Anaplasma [Line 1: 100 bp DNA marker ladder; Line
2: Ovary of cow 1; Line 3: Fetal liver; Line 4: Fetal brain; Line 5: Fetal
lung; PCR Controls; Line 6: Negative control (nuclease free water
without DNA); Line 7: Positive control (DNA of Anaplasma marginale,];
and B) Babesia [Line 1: 100 bp DNA marker ladder; Line 2: Ovary of cow
2; Line 3: Fetal lung; Line 4: Ovary of cow 2; Line 5: Fetal liver; Line 6:
Fetal brain; PCR Controls; Line 7: Positive control (DNA of Theileria
equi);
Line 8: Negative control (nuclease free water without DNA)] detection
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a haemorrhagic CL of a dark colour with a high content of leukocytes
and hemosiderin.
In the third cow, internal organ samples of one mummied embryo
were recovered, with different degrees of decomposition (FIG. 1C,
1D). A total of 30 samples of different organs (brain, lung, cardiac
muscle, skeletal muscle, among others were analysed). All samples
were negative for the presence of Brucella, Ehrlichia, Leptospira and
Toxoplasma DNA. However, DNA from members of the genus Anaplasma
and the genus Babesia was detected in brain, liver, and spleen from
the mummied foetus and in the ovaries of the two open cows (FIG 3).
The sequences generated in this study exhibited a similarity of
99% (450/453 bp) with A. marginale [GenBank accession number
MN187218] from a Bos taurus of Croatia detected in 2018. A similarity
of 99% (449/453 bp) was detected with B. bovis [MH045761] from B.
taurus of Mexico, and a 99% (493/496 pb) similarity was found with
B. bigemina [KP710228] from B. taurus of China, detected in 2012.
Anaplasma marginale was detected in all animals, whereas B. bovis
and B. bigemina were each detected in a single animal. Coinfections
with A. marginale/B. bovis and A. marginale/B. bigemina were detected
in all analysed samples.
This work now constitutes the rst record of A. marginale, B. bovis
and B. bigemina in samples from abortions and ovaries of animals
with infertility problems in Mexico. The present study is in accordance
with research carried out in a tropical region of Brazil [9], where the
presence of these pathogens was established in samples analysed
for bovine abortions. A high prevalence and low genetic diversity
of these pathogens was found in Brazil, showing that coinfections
with Anaplasma/Babesia were common, although to date, only the
coinfection of A. marginale/B. bovis has been reported. In contrast, our
study now reports the presence of A. marginale/B. bigemina in Mexican
livestock ovaries and a mummied foetus in Veracruz. These ndings
are not surprising given that both groups of pathogens are transmitted
by the same vector, the cattle tick Rhipicephalus microplus [13].
According to the histopathological results, it is important to mention
that the pathophysiology of these pathogens caused infertility [8]. It is
possible that the presence of these pathogens in the ovary triggered
a chronic inammatory process, with inltration of leukocytes and
monocytes and the generation of immunological processes, which
could have modied the physiological hormonal secretion, preventing a
correct reproductive cycle [1]. In addition, it is relevant to mention that
it would have been extremely dicult to make the diagnosis in the eld,
especially because these pathogens are not considered reproductive
pathogens. However, based on these ndings, they can be considered
as pathogens that can affect reproduction in bovine females, especially
in endemic areas of bovine anaplasmosis and babesiosis [6, 17].
CONCLUSIONS
Tick-borne pathogens represent one of the most important
challenges in livestock production worldwide. In Mexico, both bovine
anaplasmosis and bovine babesiosis cause severe economic losses
due to subclinical infections that interfere with weight gain and milk
production in animals in the productive stage. Furthermore, both
diseases generate haemolytic anaemia, which can compromise
the life of the host in acute stages. However, these pathogens are
not commonly correlated with infertility and/or abortion problems.
The question arises as to how to make the diagnosis when these
pathogens are found in the ovary or in the foetus, and it is therefore
recommended to continue with more investigations to resolve many
of the doubts that have arisen as a result of this investigation.
The ndings of the present work highlight the possibility that many
reproductive problems could be caused by thick-borne pathogens,
stressing the necessity to implement differential diagnoses of bovine
anaplasmosis and babesiosis in animals that present reproductive
problems in endemic areas of both diseases in the Mexican tropics.
Animal ethics
This study was approved by the Bioethics Committee of the
Facultad de Ciencias Biológicas y Agopecuarias, Campus Tuxpan of
the Universidad Veracruzana (UV) (Animals were handled according to
National Legislation and Ethics (NOM-012-ZOO-1993).
Tick-Borne pathogens in cows with reproductive problems / Lammoglia-Villagómez et al. _________________________________________
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ACKNOWLEDGMENTS
We thank the cattle producer and his staff for their collaboration
to get this study completed.
Financial support
The project was partially nanced by UNAM-PAPIIT IG201221.
Conicts of interest
The authors certify that they have no aliations with or involvement
in any organization or entity with any nancial interest, nonnancial
interest in the subject matter or materials discussed in this manuscript
Code availability
The sequences obtained were deposited in GenBank under
accessions numbers MZ798902 for A. marginale (16S rDNA) and
MZ798903–MZ798904 for B. bigemina and B. bovis (18S rDNA).
Ethical statement
The authors conrm that the ethical policies of the journal, as
noted on the journal's author guidelines page, have been adhered
to. This project was approved by the Bioethics and Animal Welfare
Committee of the University of Veracruz, School of Biological and
Agricultural Sciences, Tuxpan de Rodríguez Cano, Veracruz, Mexico.
BIBLIOGRAPHICS REFERENCES
[1] ALLEN, G.P. The pregnancy that doesn't stay-lessons from 25
years of observation. Vet. J. 153: 239–244. 1997. https://doi.
org/d8wmkp.
[2] ALMEIDA, A.P.; MARCILI, A.; LEITE, R.C.; NIERI-BASTOS, F.A.;
DOMINGUES, L.N.; MARTINS, J.R.; LABRUNA, M.B. Coxiella
symbiont in the tick Ornithodoros rostratus (Acari: argasidae).
Ticks. Tick. Dis. 3: 203–206. 2021. https://doi.org/f37cnr.
[3] CLOTHIER, G.; WAPENAAR, W.; KENNY, E.; WINDHAM, E. Farmers'
and veterinary surgeons' knowledge, perceptions and attitudes
towards cattle abortion investigations in the UK. Vet. Rec. Open.
187: 447. 2020. https://doi.org/grd9zx.
[4] CORREA, W.M.; CORREA, C.N.; GOTTSCHALK, A.F. Bovine
abortion associated with Anaplasma marginale. Can. J. Comp.
Med. 42: 227–228. 1978.
[5] DUGAT, T.; HACIANE, D.; DURAND, B.; LAGRÉE, A.C.; HADDAD, N.;
BOULOUIS, H.J. Identication of a Potential Marker of Anaplasma
phagocytophilum Associated with Cattle Abortion. Transbound.
Emerg. Dis. 64: e1–e3. 2017. https://doi.org/gbwj38.
[6] ESTEVE-GASENT, M.D.; RODRÍGUEZ-VIVAS, R.I.; MEDINA, R.F.;
ELLIS, D.; SCHWARTZ, A.; CORTÉS-GARCIA, B.; HUNT, C.;
TIETJEN, M.; BONILLA, D.; THOMAS, D.; LOGAN, L.L.; HASEL,
H.; ALVAREZ-MARTÍNEZ, J.A.; HERNÁNDEZ-ESCAREÑO, J.J.;
MOSQUEDA-GUALITO, J.; ALONSO-DÍAZ, M.A.; ROSARIO-CRUZ,
R.; SOBERANES-CÉSPEDES, N.; MERINO-CHARREZ, O.; HOWARD,
T.; PÉREZ DE LEÓN, A. A. Research on Integrated Management
for Cattle Fever Ticks and Bovine Babesiosis in the United States
and Mexico: Current Status and Opportunities for Binational
Coordination. Pathog. 9: 871. 2020 https://doi.org/f7fc.
[7] FOLMER, O.; BLACK, M.; HOEH, W.; LUTZ, R.; VRENHOEK, R.
DNA primers for amplication of mitochondrial cytochrome c
oxidase subunit I from diverse metazoan invertebrates. Mol.
Marine. Biol. Biotechnol. 3: 294–299. 1994.
[8] GIVENS, M.D. A clinical, evidence-based approach to infectious
causes of infertility in beef cattle. Theriogenol. 66: 648–654.
2006. https://doi.org/bmvbbp.
[9] HENKER, L.C.; LORENZETT, M.P.; FAGUNDES-MOREIRA, R.;
DALTO, A.; SONNE, L.; DRIEMEIER, D.; SOARES, J.F.; PAVARINI,
S.P. Bovine abortion, stillbirth and neonatal death associated
with Babesia bovis and Anaplasma spp. infections in southern
Brazil. Ticks. Tick. Dis. 11: 101443. 2020. https://doi.org/jzcz.
[10] HURTADO, O.J.B.; GIRALDO-RÍOS, C. Economic and health impact
of the ticks in production animals. In: Abubakar, M.; Perera,
P.K. (Eds.). Ticks and Tick-Borne Pathogens. IntechOpen 2019,
London, UK. Pp. 1–19. 2019.
[11] SARANGI, L.N.; THARANI, N.; POLAPALLY, S.; RANA, S.K.;
THODANGALA, N.; BAHEKAR, V.S.; PRASAD, A.; CHANDRASEKHAR–
REDDY, R.V.; SURENDRA, K.S.N.L.; GONUGUNTLA, H.N.; PONNANNA,
N.M.; SHARMA, G.K. Infectious bovine abortions: observations
from an organized dairy herd. Braz. J. Microbiol. 52: 439–448.
2021. https://doi.org/ghvx4g.
[12] LUCCHESE, L.; BENKIRANE, A.; HAKIMI, I.; EL IDRISSI, A.; NATALE,
A. Seroprevalence study of the main causes of abortion in dairy
cattle in Morocco. Vet. Ital. 52: 13–19. 2016. https://doi.org/jzc2.
[13] MAKENOV, M.T.; TOURE, A.H.; KORNEEV, M.G.; SACKO, N.;
PORSHAKOV, A.M.; YAKOVLEV, S.A.; RADYUK, E.V.; ZAKHAROV,
K.S.; SHIPOVALOV, A.V.; BOUMBALY, S.; ZHURENKOVA, O.B.;
GRIGOREVA, Y.E.; MOROZKIN, E.S.; FYODOROVA, M.V.; BOIRO,
M.Y.; KARAN, L.S. Rhipicephalus microplus and its vector-borne
haemoparasites in Guinea: further species expansion in West
Africa. Parasitol. Res. 120: 1563–1570. 2021. https://doi.org/mfvz.
[14] MCCALLON, B.R. Prevalence and economic aspects and
anaplasmosis: In: Proceedings of the Sixth National
Anaplasmosis Conference. Las Vegas, March 19 and 20, 1973,
Nevada, USA. Stillwater, Oklahoma: Heritage Press. Pp. 1-3. 1973.
[15] MURPHY, D.S.; LEE, X.; LARSON, S.R.; JOHNSON, D.K.; LOO, T.;
PASKEWITZ, S.M. Prevalence and distribution of human and tick
infections with the Ehrlichia muris like agent and Anaplasma
phagocytophilum in Wisconsin, 2009-2015. Vector Borne Zoon.
Dis. 17: 229–236. 2017.
[16] POTGIETER, F.T.; VAN RENSBURG, L. The persistence of colostral
Anaplasma antibodies and incidence of in utero transmission
of Anaplasma infections in calves under laboratory conditions.
Onderstepoort. J. Vet. Res. 54: 557–560. 1987.
[17] RODRÍGUEZ, S.D.; GARCÍA, O.M.A.; JIMÉNEZ, O.R.; VEGA Y
MURGUÍA, C.A. Molecular epidemiology of bovine anaplasmosis
with a particular focus in Mexico. Infect. Genet. Evol. 9: 1092–
1101. 2009. https://doi.org/dv76js.
[18] ROMERO, C.; PARDO, M.; GRILLO, M.J.; DÍAZ, R.; BLASCO,
J.M.; LÓPEZ-GOÑI, I. Evaluation of PCR and indirect enzyme-
linked immunosorbent assay on milk samples for diagnosis of
brucellosis in dairy cattle. J. Clin. Microbiol. 33: 3198–3200.
1995. https://doi.org/jzc4.
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7 of 7
[19] SUBEKTI, D.T.; FATMAWATI, M.; KHOIRIYAH, A.; PRAMESTHI,
A.; FONG, S.; DESEM, M.I.; AZMI, Z.; KUSUMANINGTYAS, E.;
ENDRAWATI, D.; PURWANTO, E.S. Seroprevalence of Seven
Reproductive Diseases in Beef and Dairy Cows from Three
Provinces in Indonesia. Vet. Med. Int. 2021(2): e6492289. 2021.
https://doi.org/jzc5.
[20] TRUEMAN, K.F.; MCLENNAN, M.W. Bovine abortion due to
prenatal Babesia bovis infection. Aus. Vet. J. 64: 63. 1987.
[21] VITAL-BRAZIL, J.M.; BALASSIANO, I.T.; OLIVEIRA, F.S.; COSTA,
A.D.; HILLEN, L.; PEREIRA, M.M. Multiplex PCR-based detection
of Leptospira in environmental water samples obtained from a
slum settlement. Mem. Inst. Oswaldo. Cruz. 105: 353–355. 2010.
https://doi.org/dn8bfp.
