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he causative agent of hemobartonellosis, a transmis- erythrocytes (1–7), causing hemolytic anemia through
sible form of anemia affecting cats, has long been extravascular destruction of erythrocytes by the
as Haemobartonella felis. This extracellular mononuclear phagocyte system and intravascular lysis
by direct damage to the cell membrane, increased
osmotic fragility, or both (7,8). Following recovery from
infection, many cats appear to harbor the organism in a
Department of Veterinary Pathology, Western College of latent carrier state (2,3,5,9).
Veterinary Medicine, University of Saskatchewan, 52 Campus This organism is fastidious, and attempts to culture it
Drive, Saskatoon, Saskatchewan S7N 5B4 (Kewish, Appleyard, in the laboratory have been unsuccessful. Based on this
Kidney, Jackson); Prairie Diagnostic Services, 52 Campus characteristic and morphology (lack of outer cell
Drive, Saskatoon, Saskatchewan S7N 5B4 (Myers).
membrane), the agent was originally classified into genus
Address all correspondence and reprint requests to Dr. Marion Haemobartonella, family Anaplasmataceae, order
Jackson; e-mail: marion.jackson@usask.ca Rickettsiales. However, recent advances in molecular
Dr. Kewish was supported by a WCVM Interprovincial Graduate techniques have demonstrated, based on nucleotide
Student Fellowship. The study was supported in part by a grant sequences of the 16S rRNA gene, that there are 2 closely
from the WCVM Companion Animal Health Fund. related species that cluster phylogenetically with
Discussion
Figure 1. Ethidium bromide-stained agarose gel (2.5%) electro- Both species of hemoplasma, M. haemofelis and
phoretogram demonstrating polymerase chain reaction (PCR) M. haemominutum, were present in blood from naturally
amplified products of mycoplasma 16S rRNA gene from periph- infected cats from Saskatchewan. The majority of cats
eral blood of cats. Lanes are as follows: A — 100 base pairs (bp) with suspected hemobartonellosis and confirmed
ladder (lowest rung is 100 bp); B and C — 2 feline samples iden- hemoplasma infection were infected with M. haemofelis.
tified as Mycoplasma haemofelis; D — Mycoplasma haemofelis Cats with normal CBCs that were PCR positive were
positive control (170 bp); E — Mycoplasma haemominutum
positive control (193 bp); F and G — 2 feline samples identified
infected with M. haemominutum and may have been
as Mycoplasma haemominutum; H — Reagent negative control. latent carriers of this organism. These results are
consistent with previous reports that M. haemominutum
is less pathogenic than M. haemofelis (13,17,18).
However, 1 cat with suspected hemobartonellosis in this
using the same primers used for PCR amplification.
study was infected with M. haemominutum. This suggests
Sequences were aligned with software (MegAlign; DNA
that, under certain circumstances, infection with
Star, Madison, Wisconsin, USA) and compared with
M. haemominutum can be associated with clinically
M. haemofelis and M. haemominutum DNA sequences by
serious disease.
using GenBank nucleotide data (accessions #AF178677,
Previous reports have suggested that cats may have
#U88563, and #U95297 for M. haemofelis; accessions
more severe anemia and clinical signs with both
#AF271154 and #U88564 for M. haemominutum).
hemobartonellosis and retroviral infection (FeLV or
FIV) than with either disease alone (19–21).
Results Hemobartonellosis typically produces a strongly
Nucleotide sequencing of 13 randomly selected amplicons regenerative anemia (3,13). However, 8 cats with NRA
confirmed the PCR products as M. haemofelis and were found to be infected with either M. haemofelis
M. haemominutum. Results of the PCR analysis of blood or M. haemominutum. These cats, by definition, were
from 3 groups of cats are presented in Table 1. In all cases, suspected to have other causes for the anemia. Of
the 2 organisms were differentiated on the basis of amplicon these 8 animals with NRA, 1 cat was FIV positive and
size following agarose gel electrophoresis (Figure 1). Within 1 was FeLV positive. In addition to exacerbating the
the group of cats with suspected hemobartonellosis, the anemia caused by hemoplasmas, concurrent retroviral
PCR was positive in 13/18 (72%); 12/18 (66%) were infection may inhibit erythroid regeneration, either
infected with M. haemofelis, and 1/18 (6%) with directly or indirectly (22). These findings emphasize
M. haemominutum. Eight of the 22 cats (36%) in the NRA/ the importance of using all available information, in
other illness group had positive PCR results. In this group, addition to the CBC, to aid in assessing the clinical
M. haemofelis was diagnosed in 4/22 (18%) and importance of hemoplasma infection. Additionally,
M. haemominutum was identified in the remaining 4. Of these findings suggest that hemobartonellosis should
the 20 cats with normal CBCs, PCR results were positive be a differential diagnosis in cats with both regenera-
in 2 (10%), both with M. haemominutum. A total of 23 of tive and nonregenerative anemias.
60 (38%) cats in this study were PCR positive, and 16 of One cat in this study had been diagnosed with hemo-
the 23 (70%) had M. haemofelis. Blood from the cat that bartonellosis 2 wk previously and was receiving
had been diagnosed with hemobartonellosis 2 wk previously doxycycline therapy at the time of PCR testing. No
and was currently receiving antibiotics was PCR negative. mycoplasma DNA was detected in the peripheral blood
The cat that had been diagnosed and treated 5 mo previously of this cat. A second cat that had been diagnosed and