Infectious blood source alters early foregut infection and regurgitative transmission of Yersinia pestis by rodent fleas

Abstract

Fleas can transmit Yersinia pestis by two mechanisms, early-phase transmission (EPT) and biofilm-dependent transmission (BDT). Transmission efficiency varies among flea species and the results from different studies have not always been consistent. One complicating variable is the species of rodent blood used for the infectious blood meal. To gain insight into the mechanism of EPT and the effect that host blood has on it, fleas were fed bacteremic mouse, rat, guinea pig, or gerbil blood; and the location and characteristics of the infection in the digestive tract and transmissibility of Y. pestis were assessed 1 to 3 days after infection. Surprisingly, 10–28% of two rodent flea species fed bacteremic rat or guinea pig blood refluxed a portion of the infected blood meal into the esophagus within 24 h of feeding. We term this phenomenon post-infection esophageal reflux (PIER). In contrast, PIER was rarely observed in rodent fleas fed bacteremic mouse or gerbil blood. PIER correlated with the accumulation of a dense mixed aggregate of Y. pestis, red blood cell stroma, and oxyhemoglobin crystals that filled the proventriculus. At their next feeding, fleas with PIER were 3–25 times more likely to appear partially blocked, with fresh blood retained within the esophagus, than were fleas without PIER. Three days after feeding on bacteremic rat blood, groups of Oropsylla montana transmitted significantly more CFU than did groups infected using mouse blood, and this enhanced transmission was biofilm-dependent. Our data support a model in which EPT results from regurgitation of Y. pestis from a partially obstructed flea foregut and that EPT and BDT can sometimes temporally overlap. The relative insolubility of the hemoglobin of rats and Sciurids and the slower digestion of their blood appears to promote regurgitative transmission, which may be one reason why these rodents are particularly prominent in plague ecology.

Author summary

Yersinia pestis, the bacterial agent of plague, is transmitted by fleas that feed on blood from rodents that carry this disease. The conclusions from studies comparing how efficiently fleas transmit plague after becoming infected have been inconsistent, possibly because a variety of rodent blood sources have been used. To investigate this, we infected three different flea species with Y. pestis using four different types of rodent blood and compared how well they could transmit three days later. The two rodent flea species that transmitted efficiently tended to reflux bacteria and blood into their esophagus when rat or guinea pig blood was used for the infections, but not when mouse or gerbil blood was used. This reflux phenomenon appears to be related to the solubility of the hemoglobin molecule of different rodent species. In contrast, cat fleas, inefficient transmitters, never refluxed their infected blood meal into the esophagus. Rodent fleas that were infected using reflux-inducing rat blood transmitted more Y. pestis than those that fed on infected mouse blood. These findings improve our understanding of how fleas transmit Y. pestis soon after becoming infected and suggest a reason why certain rodents figure more prominently in plague ecology than others.