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Background: Rickettsiae are obligate intracellular bacteria responsible for many febrile syndromes around the world, including in sub-Saharan Africa. Vectors of these pathogens include ticks, lice, mites and fleas. In order to assess exposure to flea-associated Rickettsia species in Madagascar, human and small mammal samples from an urban and a rural area, and their associated fleas were tested., Results: Anti-typhus group (TGR)- and anti-spotted fever group rickettsiae (SFGR)-specific IgG were detected in 24 (39%) and 21 (34%) of 62 human serum samples, respectively, using indirect ELISAs, with six individuals seropositive for both. Only two (2%) Rattus rattus out of 86 small mammals presented antibodies against TGR. Out of 117 fleas collected from small mammals, Rickettsia typhi, a TGR, was detected in 26 Xenopsylla cheopis (24%) collected from rodents of an urban area (n = 107), while two of these urban X. cheopis (2%) were positive for Rickettsia felis, a SFGR. R. felis DNA was also detected in eight (31%) out of 26 Pulex irritans fleas., Conclusions: The general population in Madagascar are exposed to rickettsiae, and two flea-associated Rickettsia pathogens, R. typhi and R. felis, are present near or in homes. Although our results are from a single district, they demonstrate that rickettsiae should be considered as potential agents of undifferentiated fever in Madagascar.

Louse borne typhus (also called epidemic typhus) was one of man's major scourges, and epidemics of the disease can be reignited when social, economic, or political systems are disrupted. The fear of a bioterrorist attack using the etiologic agent of typhus, Rickettsia prowazekii, was a reality. An attenuated typhus vaccine, R. prowazekii Madrid E strain, was observed to revert to virulence as demonstrated by isolation of the virulent revertant Evir strain from animals which were inoculated with Madrid E strain. The mechanism of the mutation in R. prowazekii that affects the virulence of the vaccine was not known. We sequenced the genome of the virulent revertant Evir strain and compared its genome sequence with the genome sequences of its parental strain, Madrid E. We found that only a single nucleotide in the entire genome was different between the vaccine strain Madrid E and its virulent revertant strain Evir. The mutation is a single nucleotide insertion in the methyltransferase gene (also known as PR028) in the vaccine strain that inactivated the gene. We also confirmed that the vaccine strain E did not cause fever in guinea pigs and the virulent revertant strain Evir caused fever in guinea pigs. We concluded that a single nucleotide insertion in the methyltransferase gene of R. prowazekii attenuated the R. prowazekii vaccine strain E. This suggested that an irreversible insertion or deletion mutation in the methyl transferase gene of R. prowazekii is required for Madrid E to be considered a safe vaccine.

Epidemic typhus, caused by Rickettsia prowazekii, is maintained in a southern flying squirrel (Glaucomys volans) sylvatic cycle in the southeastern United States. The northern flying squirrel (Glaucomys sabrinus) has not been previously associated with R. prowazekii transmission. A second rickettsial pathogen, Anaplasma phagocytophilum, infects dusky-footed woodrats (Neotoma fuscipes) and tree squirrels in northern California. Because northern flying squirrels or their ectoparasites have not been tested for these rickettsial pathogens, serology and polymerase chain reaction (PCR) were used to test 24 northern flying squirrels for R. prowazekii and A. phagocytophilum infection or antibodies. Although there was no evidence of exposure to R. prowazekii, we provide molecular evidence of A. phagocytophilum infection in one flying squirrel; two flying squirrels also were seropositive for this pathogen. Fleas and ticks removed from the squirrels included Ceratophyllus ciliatus mononis, Opisodasys vesperalis, Ixodes hearlei, Ixodes pacificus, and Dermacentor paramapertus.

Genetic traits of five Rickettsia prowazekii isolates, including the first from Africa and North America, and representatives from human and flying squirrels were compared using multilocus sequence typing. Four rickettsial genes encoding 17 kDa genus-common antigen (17 kDa gene), citrate synthase (gltA), OmpB immunodominant antigen (ompB) and 120 kDa cytoplasmic antigen (sca4) were examined. Sequence identities of 17 kDa gene and gltA were 100% among the isolates. Limited sequence diversity of ompB (0.02-0.11%) and sca4 (0.03-0.20%) was enough to distinguish the isolates, and evaluation of the combined four genes provided a method to easily differentiate R. prowazekii from other rickettsiae.

The first description of Mediterranean spotted fever (MSF) was made by Conor and Brush in 1910 in Tunisia, where, at the same time, Nicolle described the role of lice in transmission of epidemic typhus. However, along this century, there have been few and fragmentary reports about ecology and epidemiology of rickettsioses in North Africa. This region was always considered, for these diseases, like other Mediterranean regions. The most human tick-borne rickettsiosis known to occur in North Africa is MSF caused by R. conorii and transmitted by the brown dog tick, Rhipicephalus sanguineus. Recent studies showed that other arthropode-transmitted rickettsiae are prevalent in North Africa: R. aeschlimannii, R. massiliae, and R. felis. Moreover, R. felis and R. aeschlimannii human infection were respectively confirmed, by serology in Tunisia, and by PCR in Morocco. The seroprevalence of R. conorii among healthy population was ranging from 5% to 8% in most of the countries. Epidemiological and clinical features are frequently resumed in an eruptive fever with eschar occurring in hot season in rural areas. Typhus group rickettsioses, caused by R. typhi and R. prowazekii are less frequently reported than in the 1970s. Seroprevalence of R. typhi among blood donors was from 0.5% to 4%. In Algeria about 2% of febrile patients had R. prowazekii antibodies. Moreover, reemerging threat of epidemic typhus should be considered, after the two cases recently diagnosed in the highlands of Algeria. Murine typhus, considered as "benign" typhus, is underestimated. When R. typhi was inserted in serologic tests, murine typhus became more frequently confirmed. In a recent study in Central Tunisia, we confirmed an emergence of murine typhus mistaken for R. conorii or viral infection. In addition to typhus surveillance, future studies have to determine which spotted fever group rickettsiae are prevalent in vectors and in human pathology.

The authors infected lambs with R. prowazeki and R. canada to ascertain their possible role in the natural infection of the animals. The lambs were infected subcutaneously with increasing doses; rickettsiemia was recorded with the aid of tests on guinea pigs and Ixodidae and Argasidae ticks fed on the lambs. Dynamics of antibody formation was ascertained in the infected animals in the agglutination reaction and in the complement fixation test. The antigenic affinity of R. canada and rickettsia of the typhoid group and the presence of common antigenic determinants with the Proteus OX19 was confirmed. The absence of any clinical manifestations, the character of antibody formation, impossibility of inducing the generalized infection and of the isolation of the causative agent from the blood pointed to the low susceptibility of lambs to R. prowazeki and R. canada; thus a possibility of circulation of the causative agents of typhius among the domestic animals scarcely probable.

Typhus fever has occurred globally as epidemic and endemic disorders. In 1910, Brill reported a typhus-like illness which Zinsser and others determined to be recurrent epidemic typhus fever. Maxcy, in 1926, proposed rodents and fleas as reservoir and vector, respectively, of endemic typhus, which Dyer confirmed in 1930. Animals experimentally infected with epidemic typhus (Rickettsia prowazeki) are immune to murine typhus (Rickettsia typhi) and vice versa. Similar solid cross-immunity exists for humans. The two diseases are clinically similar in pathologic and serologic reactions. Human epidemic typhus presumably involved a man-louse-man cycle without an animal reservoir. This concept is now questioned. Antibodies to R. prowazeki have been reported in livestock in Africa, rats in Manila, and from flying squirrels and humans in the United States. R. prowazeki was recovered from blood specimens of goats, sheep, from ixodid ticks, louse, and flea-ectoparasites of flying squirrels, and tissues of flying squirrels. More than 20 cases of squirrel-related acute epidemic typhus have been reported in the United States. R. prowazeki has not been recovered from human cases. Chemical studies of R. prowazeki and R. typhi show genetic similarities but differences in genome size and degree of hybridization suggest that interconversions between the two agents do not occur rapidly in nature. It is proposed that, with time, their relatedness will become even closer.

Epizootiologic studies conducted during the past few years showed the existence of widespread natural infection of the southern flying squirrel, Glaucomys volans, with epidemic typhus rickettsiae, Rickettsia prowazekii. The ecological findings strongly implicated transmission of the etiologic agent by an arthropod vector. Studies were conducted under controlled laboratory conditions to determine whether ectoparasites naturally associated with flying squirrels (squirrel fleas, lice, mites and ticks) were capable of acquiring, maintaining and transmitting the infection. Also studied were the cat flea, oriental rat flea and the human body louse. Flying squirrels inoculated with the GvF-16 strain of R. prowazekii circulated rickettsiae in their blood for 2-3 weeks, thus providing ample opportunity for arthropods feeding on them to become infected. The results with Dermacentor variabilis ticks indicated that the rickettsiae did not consistently survive in this insect and were not passed to the eggs of adult females that had been infected subcuticularly. Mites became infected by feeding on infectious blood but failed to sustain the infection. Also, mites fed on an infected flying squirrel did not transmit the infection to a normal squirrel. Squirrel, cat, and oriental rat fleas readily became infected by feeding on a rickettsemic host or on infectious blood through membranes, but failed to transmit the infection to susceptible flying squirrels. In the studies with flying squirrel lice, however, transmission of epidemic typhus from infected to uninfected flying squirrels was demonstrated. Infection of the human body louse with the GvF-16 flying squirrel strain of R. prowazekii was similar to that previously observed with classical human strains, viz., multiplication of the rickettsiae and excretion in the feces.

Previous work suggested that domestic animals could act as a reservoir of epidemic typhus. To test this hypothesis, sera from buffaloes, camels, donkeys, goats, pigs and sheep in different parts of Egypt were submitted to the complement-fixation test, using the common soluble typhus antigen. The highest percentage of positive results was obtained with camel and donkey sera and the positivity rate was higher towards the end of the year than in June and July. Animals from the Western Desert area were more frequently positive than those from other regions. Attempts to isolate rickettsiae by intraperitoneal injection of guinea-pigs resulted in some febrile reactions, but so far no rickettsiae have been isolated.