Abstract
Abstract
Background
Feline-associated hemotropic
Mycoplasma
(hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (
Ctenocephalides felis
). While the efficiency of transmission by
C. felis
appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low ( 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on
C. felis
hemoplasma infection prevalence.
Methods
A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on
C. felis
collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5
C. felis
each, half of which were washed and half not washed.
Results
Flea washing did not influence the detection of hemoplasma but instead amplified
Spiroplasma
. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C.
felis
samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining “hemoplasma infected” fleas, PCR amplified
Spiroplasma
or other bacteria.
Conclusions
Therefore, we concluded that hemoplasma infection in
C. felis
is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of
C. felis
to maintain hemoplasma infection is necessary.