Identification of serum biomarkers in dogs naturally infected with Babesia canis canis using a proteomic approach

Erythropoietin (Epo) is produced primarily in the kidneys upon low blood oxygen availability and stimulates erythropoiesis in the bone marrow. Recombinant human Epo (rHuEpo), a drug developed to increase arterial oxygen content in patients, is also illicitly used by athletes to improve their endurance performance. Therefore, a robust and sensitive test to detect its abuse is needed. The aim of the present study was to investigate potential human serum biomarkers of Epo abuse employing a proteomic approach. Eight healthy male subjects were injected subcutaneously with rHuEpo (5,000 IU) every second day for a 16-day period. Serum was collected before starting the treatment regime and again at days 8 and 16 during the treatment period. Samples were homogenized and proteins separated by two-dimensional gel electrophoresis (2DE). Spots that changed significantly in response to rHuEpo treatment were identified by mass spectrometry. Both the number of reticulocytes and erythrocytes increased throughout the study, leading to a significant increase in hematocrit and hemoglobin content. In addition, transferrin levels increased but the percentage of iron bound to transferrin and ferritin levels decreased. Out of 97 serum proteins, seven were found to decrease significantly at day 16 compared with pre-Epo administration, and were identified as four isoforms of haptoglobin, two isoforms of transferrin, and a mixture of hemopexin and albumin. In support, total serum haptoglobin levels were found to be significantly decreased at both days 8 and 16. Thus a 2DE proteomic approach for discovery of novel markers of Epo action appears feasible.

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Molecular detection of co-infections with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofilaria-positive dogs from Slovakia

Veterinary Parasitology ◽

10.1016/j.vetpar.2014.01.022 ◽

2014 ◽

Vol 203 (1-2) ◽

pp. 167-172 ◽

Cited By ~ 17

Author(s):

Bronislava Víchová ◽

Martina Miterpáková ◽

Adriana Iglódyová

Keyword(s):

Anaplasma Phagocytophilum ◽

Molecular Detection ◽

Babesia Canis ◽

Babesia Canis Canis

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The prevention of transmission of Babesia canis canis by Dermacentor reticulatus ticks to dogs using a novel combination of fipronil, amitraz and (S)-methoprene

Veterinary Parasitology ◽

10.1016/j.vetpar.2011.03.047 ◽

2011 ◽

Vol 179 (4) ◽

pp. 343-350 ◽

Cited By ~ 30

Author(s):

Frans Jongejan ◽

Josephus J. Fourie ◽

S. Theodore Chester ◽

Coralie Manavella ◽

Yasmina Mallouk ◽

...

Keyword(s):

Dermacentor Reticulatus ◽

Babesia Canis ◽

Babesia Canis Canis

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The loop-mediated isothermal amplification assay for rapid diagnosis of Babesia canis canis infections in dogs

Polish Journal of Veterinary Sciences ◽

10.2478/pjvs-2013-0020 ◽

2013 ◽

Vol 16 (1) ◽

pp. 131-133 ◽

Cited By ~ 1

Author(s):

Ł. Adaszek ◽

M. Jankowska ◽

M. Kalinowski ◽

T. Banach ◽

D. Wułupek ◽

...

Keyword(s):

High Specificity ◽

Isothermal Amplification ◽

Lamp Method ◽

Babesia Canis ◽

Canine Babesiosis ◽

Loop Mediated Isothermal Amplification ◽

Specificity And Sensitivity ◽

Amplification Assay ◽

Analytical Laboratories ◽

Babesia Canis Canis

Abstract The aim of this study was to use a rapid and easy DNA-based test, the loop-mediated isothermal amplification (LAMP), for diagnosis of Babesia canis canis infections in dogs. 10 DNA samples of 18S RNA-A and 10 DNA samples of 18S RNA-B of B. canis canis were used in the study. LAMP method could successfully detect DNA in all examined samples down to 0.1 pg dilution. Obtained results suggest that this method has high specificity and sensitivity and can be applied in analytical laboratories in diagnosis of canine babesiosis.

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Molecular evidence of natural infection with Babesia canis canis in Croatia

Acta Veterinaria Hungarica ◽

10.1556/avet.58.2010.1.4 ◽

2010 ◽

Vol 58 (1) ◽

pp. 39-46 ◽

Cited By ~ 7

Author(s):

Mirna Brkljačić ◽

Vesna Matijatko ◽

Ivana Kiš ◽

Nada Kučer ◽

Jadranka Foršek ◽

...

Keyword(s):

Sequence Analysis ◽

Genetic Variant ◽

Natural Infection ◽

Molecular Evidence ◽

Rrna Gene ◽

Babesia Canis ◽

Nucleotide Difference ◽

Dark Urine ◽

Blood Smears ◽

Babesia Canis Canis

The aim of the present study was to detect and characterise the species and subspecies of Babesia spp. that cause canine babesiosis in Croatia. Twenty-eight dogs with typical signs of babesiosis (lethargy, anorexia, fever, dark urine and thrombocytopenia) were included in this study. Their blood smears showed the presence of Babesia canis . The results showed the detection of one subspecies, namely Babesia canis canis using PCR, and subsequent sequence analysis demonstrated portions of the nss rRNA gene in 27 out of 28 samples. Sequence analysis of the isolates showed 100% identity in 11 samples, 99.7% identity (one nucleotide difference) in 11 samples and 99.4% identity (two nucleotides difference) in 5 samples with B. canis canis . The results of this study confirm the presence of B. canis canis in infected dogs in Croatia and demonstrate a slightly new genetic variant of Babesia subspecies.

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Postmortem small babesia-like morphology of Babesia canis — Short communication

Acta Veterinaria Hungarica ◽

10.1556/avet.2011.029 ◽

2011 ◽

Vol 59 (4) ◽

pp. 427-432 ◽

Cited By ~ 7

Author(s):

Zoltán Demeter ◽

Elena Palade ◽

Éva Balogh ◽

Csaba Jakab ◽

Róbert Farkas ◽

...

Keyword(s):

Causative Agent ◽

18S Rrna Gene ◽

Severe Anaemia ◽

Rrna Gene ◽

Babesia Canis ◽

Post Mortem ◽

Tissue Samples ◽

Tissue Sections ◽

Sequence Identity ◽

Babesia Canis Canis

Here we report a case of canine babesiosis with unusual morphology of the causative agent. A male, seven-week-old Labrador retriever puppy, exhibiting severe anaemia and haemoglobinuria, was presented at the Clinic of Internal Medicine in February 2011. The puppy was euthanised. The most relevant pathological changes were icterus, severe splenomegaly, generalised lymphadenopathy and haemoglobin nephrosis. Samples were collected from various organs for histology within one hour post mortem. Impression smears were also prepared from the spleen after overnight storage at 4 °C. Tissue sections and smears showed the presence of multiple, coccoid intraerythrocytic bodies that measured 1–2 μm and resembled small babesiae. No large piroplasms were seen. DNA was extracted from the spleen, and a conventional PCR was performed for the amplification of a 450-bp region of the 18S rRNA gene of piroplasms. The causative agent was identified as Babesia canis canis, with 99% sequence identity to other European isolates. Sequence identity to B. gibsoni was only 91%. This is the first account to verify that the morphology of the large canine piroplasm, B. canis, can be uniformly small babesia-like post mortem or following the storage of tissue samples.

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