A survey of the vectors of spotted fever group Rickettsiae and of murine typhus was carried out in Rahat, a Bedouin town in the Negev Desert, where the diseases are endemic. Houses with known cases of spotted fever group Rickettsiae or murine typhus were compared with those without reported clinical cases. A neighboring Jewish community, Lehavim, where no cases of spotted fever group Rickettsiae and murine typhus were reported in recent years, was used as a control. In the houses of patients with spotted fever group Rickettsiae in Rahat, an average of 7.4 times more ticks were found than in control houses. Out of 190 ticks isolated from sheep and goats or caught by flagging in Rahat, 90% were Rhipicephalus sanguineus, 7.9% Rhipicephalus turanicus, and 2.1% were Hyalomma spp. In the houses of patients with murine typhus, three times more rats were caught and, on the average, each rat was infested with 2.2 times more fleas than rats in the control houses. Out of 323 fleas collected from 35 Norwegian rats (Rattus norvegicus), 191 were Xenopsylla cheopis and 132 Echidnophaga murina. Thus, there was a six to seven times higher probability of encountering a tick or flea vector where infections had occurred than in control houses in Rahat. The percentage of rats seropositive to Rickettsia typhi was similar in study and control households (78.3 and 76.2, respectively). In the control settlement, Lehavim, only three Mus musculus were caught, which were not infested with ectoparasites and their sera were negative for murine typhus. Out of 10 dogs examined in this settlement, 15 R. sanguineus and eight specimens of the cat flea (Ctenocephalides felis felis) were isolated. No rats were caught in this settlement. These data indicate that there is a correlation among the density of domestic animals, their ectoparasites, and the incidence of spotted fever group Rickettsiae and murine typhus in Rahat (Mumcuoglu et al. 2001).

The prevalence of Bartonella spp. in wild rodents was studied in 19 geographical locations in Israel. One hundred and twelve rodents belonging to five species (Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus) were included in the survey. In addition, 156 ectoparasites were collected from the rodents. Spleen sample from each rodent and the ectoparasites were examined for the presence of Bartonella DNA using high resolution melt (HRM) real-time PCR. The method was designed for the simultaneous detection and differentiation of eight Bartonella spp. according to the nucleotide variation in each of two gene fragments (rpoB and gltA) and the 16S-23S intergenic spacer (ITS) locus, using the same PCR protocol which allowed the simultaneous amplification of the three different loci. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). In addition, 15 of 34 (44%) flea pools harbored Bartonella DNA. Only rat flea (Xenopsyla cheopis) pools collected from black rats (R. rattus) were positive for Bartonella DNA. The Bartonella sp. detected in spleen samples from black rats was closely related to both B. tribocorum and B. elizabethae. The species detected in the Cairo spiny mouse spleen sample was closely related to the zoonotic pathogen, B. elizabethae. These results indicate that Bartonella species are highly prevalent in suburban rodent populations and their ectoparasites in Israel (Morick et al. 2010).

Out of 340 stray cats in Jerusalem, 186 (54.7%) were infested with the cat flea, Ctenocephalides felis, 49 (14.4%) with the cat louse, Felicola subrostratus, 41 (12.0%) with the ear mite, Otodectes cynotis, three (0.9%) with the fur mite, Cheyletiella blakei, two (0.6%) with the itch mite Notoedres cati, and 25 (7.3%) with ticks of the species Rhipicephalus sanguineus sl, Rhipicephalus turanicus or Haemaphysalis adleri. A higher number of flea infestations was observed in apparently sick cats (P <0.05) and in cats aged <6 months (P <0.05). The proportion of flea-infested cats (P <0.01), as well as the number of fleas per infested cat (P <0.01), was higher in autumn than in other seasons. By contrast with findings in cats with flea infestations, rates of infestation with ticks were higher amongst cats with clinical signs (P <0.01) and cats aged ≥6 months (P <0.05) (Salant et al. 2013).

The aim of the current study was to investigate the presence of Bartonella spp. in commensal rodents and their ectoparasites in Nigeria. We report, for the first time, the molecular detection of Bartonella in 26% (46/177) of commensal rodents (Rattus rattus, R. norvegicus and Cricetomys gambianus) and 28% (9/32) of ectoparasite pools (Xenopsylla cheopis, Haemolaelaps spp., Ctenophthalmus spp., Hemimerus talpoides, and Rhipicephalus sanguineus) from Nigeria. Sequence analysis of the citrate synthase gene (gltA) revealed diversity of Bartonella spp. and genotypes in Nigerian rodents and their ectoparasites. Bartonella spp. identical or closely related to Bartonella elizabethae, Bartonella tribocorum and Bartonella grahamii were detected. High prevalence of infection with Bartonella spp. was detected in commensal rodents and ectoparasites from Nigeria. The Bartonella spp. identified, were previously associated with human diseases highlighting their importance to public health (Kamani et al. 2013).

The prevalence and genetic characteristics of Bartonella spp. in fleas of wild and domestic animals from Palestinian territories are described. Flea samples (n=289) were collected from 121 cats, 135 dogs, 26 hyraxes and seven rats from northern (n=165), central (n=113), and southern Palestinian territories (n=11). The prevalent flea species were: Ctenocephalides felis (n=119/289; 41.2%), Ctenocephalides canis (n=159/289; 55%), and Xenopsylla sp. (n=7/289; 2.4%). Targeting the Intergenic Transcribed Spacer (ITS) locus, DNA of Bartonella was detected in 22% (64/289) of all fleas. Fifty percent of the C. felis and 57% of the Xenopsylla sp. contained Bartonella DNA. DNA sequencing showed the presence of Bartonella clarridgeiae (50%), Bartonella henselae (27%), and Bartonella koehlerae (3%) in C. felis. Xenopsylla sp. collected from Rattus rattus rats were infected with Bartonella tribocorum, Bartonella elizabethae, and Bartonella rochalimae. Phylogenetic sequence analysis using the 16S ribosomal RNA gene obtained four genetic clusters, B. henselae and B. koehlerae as sub-cluster 1, B. clarridgeiae as cluster 2, while the rat Bartonella species (B. tribocorum and B. elizabethae) were an outgroup cluster (Nasereddin et al. 2014).

The characterization of flea species infesting kennel dogs from two localities in Israel (Rehovot and Jerusalem) and their molecular screening for Bartonella species was investigated. A total of 355 fleas were collected from 107 dogs. The fleas were morphologically classified and molecularly screened targeting the Bartonella 16S–23S internal transcribed spacer (ITS). Of the 107 dogs examined, 80 (74.8%) were infested with Ctenocephalides canis, 68 (63.6%) with Ctenocephalides felis, 15 (14.0%) with Pulex irritans and one (0.9%) with Xenopsylla cheopis. Fleas were grouped into 166 pools (one to nine fleas per pool) according to species and host. Thirteen of the 166 flea pools (7.8%) were found to be positive for Bartonella DNA. Detected ITS sequences were 99–100% similar to those of four Bartonella species: Bartonella henselae (six pools); Bartonella elizabethae (five pools); Bartonella rochalimae (one pool), and Bartonella bovis (one pool) (Sofer et al. 2015).

Four hundred and sixty seven Ctenocephalides felis fleas removed from185 feral cats living in residential areas of Jerusalem, Israel, were screened for bacterial infections of public health importance. The fleas were screened for bartonellae, rickettsiae and Coxiella burnetii by PCR and sequencing. Bartonella DNA was detected in 156 individual fleas collected from 91 of the 185 (49.2%) cats. DNA of Bartonella clarridgeiae, Bartonella henselae and Bartonella koehlerae was detected in 112/467 (24%), 29/467 (6.2%) and 15/467 (3.2%), respectively, indicating a significantly different distribution of these Bartonella spp. among the fleas. However, no differences were observed between female and male fleas in their Bartonella-infection status. Ninety one individual cats carried fleas infected with 1 to 3 Bartonella species. No differences were found between fleas collected from male and female, pregnant and non-pregnant or young, juvenile and adult cats. Interestingly, a significant association was observed between the clinical status of the cat hosts (apparently healthy versus sick) and the carriage of Bartonella-positive fleas. One of the 467 (0.2%) fleas was positive for Rickettsia felis DNA and no other Rickettsia spp. or C. burnetii DNA were detected (Kamani et al. 2015).

Molecular assays were used to detect and characterize two agents of zoonotic importance, Coxiella burnetii and Rickettsia spp. in 194 peridomestic rodents captured in a peri-urban setting in Nigeria, and 32 pools of ectoparasites removed from them, to determine their possible role in the epidemiology of these diseases in this country. Targeting and characterizing the insertion sequence IS1111, C. burnetii DNA was detected in 4 out of 194 (2.1%) rodents comprising 3 out of 121 (2.5%) Rattus norvegicus and 1 out of 48 (2.1%) Rattus rattus screened in this study. Rickettsia spp. DNA was detected in two Rhipicephalus sanguineus sensu lato pools (i.e. RT1 and RT4) using the citrate synthase (gltA) gene and further characterized by amplification and sequence analysis of six genes to determine their identity. The RT1 sample consistently gave 98-100% identity to Rickettsia conorii str. Malish 7 for the various genes and loci studied. Five pools of Xenopsylla cheopis and Ctenophthalmus spp. were negative (Kamani et al. 2018a).

This study was conducted to determine the prevalence of pathogenic and endosymbiont apicomplexans in the cat flea, Ctenocephalides felis (Bouché) infesting 185 stray cats in Jerusalem, Israel using PCR assay and sequencing approach. Two pathogens, Hepatozoon felis and Babesia vogeli and an endosymbiont Steinina ctenocephali were detected in 1.9%, 0.2% and 5.8% of 685 C. felis evaluated respectively. There was a significant association (p < 0.05) between the prevalence of H. felis and the sex of cats hosting the fleas as well as the season of sampling but not for age or health status of the cats or sex of the fleas tested. Prevalence of S. ctenocephali was significantly (p < 0.001) associated with season, being higher in the warm season. This report represents the first molecular detection of S. ctenocephali in C. felis (Kamani et al. 2018b).

Kamani, J, Morick D, Mumcuoglu KY, Harrus S. 2013. Prevalence and diversity of Bartonella species in commensal rodents and ectoparasites from Nigeria, West Africa. PLoS Neg Trop Dis 7 (5): e2246. doi:10.1371/journal.pntd.0002246.

Kamani J, Baneth G, Apanaskevich DA, Mumcuoglu KY, Harrus S. 2015. Molecular detection of Rickettsia aeschlimannii in Hyalomma spp. ticks from camels (Camelus dromedarius) in Nigeria, West Africa. Med. Vet. Entomol. doi: 10.1111/mve.12094.

Kamani J, Baneth G, Gutiérrez R, Nachum-Biala Y, Mumcuoglu KY, Harrus S. 2018a. Coxiella burnetii and Rickettsia conorii: Two zoonotic pathogens in peridomestic rodents and their ectoparasites in Nigeria. Ticks Tick Borne Dis. 2017 Oct 12. pii: S1877- 959X(17)30242-X. doi: 10.1016/j.ttbdis.2017.10.004.

Kamani J, Harrus S, Nachum-Biala Y, Salant H, Mumcuoglu KY, Baneth G. 2018b. Pathogenic and endosymbiont apicomplexans in Ctenocephalides felis (Siphonaptera: Pulicidae) from cats in Jerusalem, Israel. Comp. Immunol. Microbiol. Infect. Dis. 57: 29- 33. https://doi.org/10.1371/journal.pntd.0006395.

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Nasereddin A, A. Risheq, S. Harrus, K. Azmi, S. Ereqat, G. Baneth, H. Salant, K.Y. Mumcuoglu & Z. Abdeen. 2014. Bartonella species in fleas from Palestinian territories: Prevalence and genetic diversity. Journal of Vector Ecology 39 (2): 261-270.

Salant, H., Mumcuoglu, K.Y., Baneth, G. 2013. Ectoparasites in urban stray cats in Jerusalem, Israel: differences in infestation patterns of fleas, ticks and permanent ectoparasites. Med. Vet. Entomol. 28: 314-318, doi: 10.1111/mve.12032.

Sofer S, Gutiérrez R, Morick D, Mumcuoglu KY, Harrus S. 2015. Molecular detection of zoonotic Bartonella (B. henselae, B. elizabethae and B. rochalimae) in fleas collected from dogs in Israel. Med. Vet. Entomol. 2015; 29(3): 344-8. doi: 10.1111/mve.12116.

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