Rickettsia typhi, Bartonella henselae and related zoonotic agents in fleas from domestic cats (Felis catus) from the Rio Grande Valley, Texas

Rickettsia typhi, Bartonella henselae and related zoonotic agents in fleas from domestic cats (Felis catus) from the Rio Grande Valley, Texas | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Rickettsia typhi, Bartonella henselae and related zoonotic agents in fleas from domestic cats (Felis catus) from the Rio Grande Valley, Texas Sujata Balasubramanian, Italo Zecca, Allyson Koger, Sarah Hamer This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8726915/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Background Flea-borne rickettsiosis and bartonellosis are emerging diseases in humans and animals. There is increasing concern for the emergence of Rickettsia typhi (agent of murine typhus or flea-borne typhus) in humans of southern USA, for which cats are key reservoirs and cat fleas ( Ctenocephalides felis ) serve as the vector. Similarly, multiple species of Bartonella have cats as natural hosts and fleas as vectors, leading to varied disease outcomes in humans, including cat scratch disease and neurobartonellosis. Methods To investigate the ecology of Rickettsia and Bartonella pathogens, we studied 167 predominantly stray cats ( Felis catus ) from the Rio Grande Valley in south Texas. Fleas were collected, identified morphologically and confirmed molecularly. DNA from fleas was tested for Rickettsia and Bartonella using PCR and Sanger sequencing of multiple gene targets. Additionally, DNA from blood of all cats and tissues from a subset of cats were assayed for infections. Results Flea infestation prevalence of cats was 83% - higher in male vs. female cats - yielding 721 fleas, predominantly C. felis with 2 Echidnophaga gallinacea and 1 Pulex sp . Flea burden was a significant predictor of Rickettsia infection in fleas. Candidatus Rickettsia senegalensis was identified in 28 flea pools (20% of all pools) and Rickettsia typhi in 6 flea pools (4%). Bartonella henselae was identified in 59 flea pools (42% of all pools) and blood from 37 cats (22.2%); Bartonella clarridgeiae was detected in 3 flea pools (2% of pools) and blood from 1 cat (0.7%). Thirty cats had simultaneously Bartonella- positive fleas and blood. Rickettsia infection was positively associated with flea burden. Co-infections of Rickettsia and Bartonella in fleas occurred more commonly than expected by chance. Conclusions Cats and their fleas support robust transmission of R. typhi , B. henselae and related agents in south Texas with implications for veterinary and public health. Ctenocephalides felis Rickettsia typhi murine typhus Bartonella henselae cat scratch disease Rio Grande Valley Texas Figures Figure 1 Background Flea-borne diseases continue to emerge in human populations worldwide, with increasing attention to pathogens in the genera Rickettsia and Bartonella . At least 17 species of Rickettsia cause disease in humans, many resulting in spotted fever or typhus fever ( 1 , 2 ). Rickettsia typhi – the causative agent of murine typhus or flea-borne typhus - is an emergent global threat for humans ( 2 – 4 ). When typhus was initially identified in the 1920s, rats were the reservoirs, and the rat flea ( Xenopsylla cheopis) was the recognized vector. R. typhi is now also transmitted by Ctenocephalides felis (cat flea) and other flea species ( 5 ), with opossums ( Didelphis virginiana ) and cats ( Felis catus ) as reservoirs ( 5 , 6 ). R. typhi is endemic to south Texas, California, and Hawaii in the US. Infection with R. typhi may cause fever, headache, chills, rash, gastrointestinal or renal symptoms with potential for hospitalization, though mortality is rare and treatment is accomplished with antibiotics ( 7 ). Clinical impacts on infected pets are not well studied, although infected cats have been reported from many human disease hotspots, and one case study identified R. typhi in a clinically ill dog in Texas ( 8 – 10 ). Multiple species of Bartonella are recognized as emerging threats to public health ( 11 ). Bartonella henselae is the main causal agent of cat scratch disease is but B. clarridgeiae is also considered a causal agent ( 12 , 13 ). In humans, infection with Bartonella has different outcomes for immunocompetent and immunocompromised human hosts ( 14 ). Humans acquire bartonellosis from bites or scratches from cats, more frequently diagnosed in children. Cat-scratch disease in humans progresses acutely from skin lesion to fever and lymphadenopathy. Neurological complications, often ocular, have been observed with Bartonella spp infections ( 15 – 17 ). In cats, B. henselae is maintained in an intra-erythrocytic state and typically results in asymptomatic bacteremia ( 14 , 18 ), although infected cats may develop cardiovascular issues including lymphoplasmacytic myocarditis among other ailments affecting the lymph nodes, spleen, liver, and kidneys; sometimes fatal ( 19 , 20 ). The cat flea ( C. felis ) is a predominant vector for R. typhi , B. hensela e and related species. This flea has a worldwide distribution and is an abundant and frequent ectoparasite on domestic dogs and cats. Across broad geographic ranges, C. felis is the most common flea species found on domestic and feral cats, often among other species including the sticktight flea ( Echidnophaga gallinacea ) and human flea ( Pulex irritans ) ( 21 – 25 ). In addition to Rickettsia and Bartonella spp. pathogens, C. felis may transmit or carry Yersinia pestis (plague), Dipylidium caninum (tapeworm), Hymenolepis diminuta (rat tapeworm), Dipetalonema ( Acanthocheilonema ) reconditum and hemoplasmas ( 26 – 28 ), and may directly impact hosts through flea allergic dermatitis. Considering the emergent threat of R. typhi and B. henselae in the U.S., our work aims to describe the prevalence of both genera among fleas and their cat hosts in the Rio Grande Valley in south Texas. Methods The Rio Grande Valley of south Texas is a region with a subtropical climate that supports many vector species and tropical infections. We partnered with a large shelter in Hidalgo County that intakes an estimated 9,000 cats per year. Cats in our study included 167 predominantly stray cats from 14 surrounding rural and urban zones of the Rio Grande Valley, and were the subject of a prior study of Trypanosoma cruzi and feline Chagas disease ( 29 ). These cats were sampled across winter (01 Jan-19 Mar), spring (20 Mar − 20 June) and summer (21 June – 19 Sept) of 2017. At the time of euthanasia for non-study reasons, the sex, and geographic origin of cats was recorded. Cats were combed using flea combs across the whole body for approximately three minutes and all recovered fleas were preserved in 70% ethanol. As described previously ( 29 ), blood was collected from all cats, spun and separated in the laboratory with DNA extracted from the blood clot available for the current study. Additionally, DNA extracted from heart was available from all cats and other tissues were also available from cats sampled in spring and summer. Fleas were identified using the key from Centers for Disease Control and Prevention with pictorial details from Lawrence et al. ( 30 , 31 ). When ≤5 conspecific fleas were collected from an individual cat, they were pooled for DNA extraction. When > 5 conspecific fleas were present, 5 were randomly selected to create a pool. When > 1 flea species was present, a pool was made for each species. Representative fleas have been deposited into Texas A&M University Insect Collection (TAMUIC). The Omega EZNA (Omega Bio-tek, Norcross, GA) kit was used to extract DNA from fleas. Fleas were first crushed into lysis buffer using single-use pestles. To confirm the flea species identification, representatives of each species were subjected to a molecular identification process using PCR targeting the insect cytochrome oxidase I locus with primers LCO1490 and HC02198 (Additional file 1: Table S1 ) ( 32 ). PCR products were sequenced, and chromatograms were analyzed as described below. Representative flea sequences were deposited to Genbank (PP545303-PP545306). Three different PCRs were used to identify Rickettsia (Additional file 1: Table S1 ) from flea pools and selected cat blood clots and tissues. Outer membrane protein ( rOmpB ) was amplified using primers 120–2788 and 120–3599; this assay can detect a wide range of Rickettsial species ( 33 ). Two sets of primers were used to amplify the gltA (citrate synthase) gene: RrCS to detect spotted fever group Rickettsia using primers RrCS 372 and RrCS 989 ( 34 ) and RpCS to detect the typhus group Rickettsia using primers RpCS 877 and 1258n ( 35 ). For the blood and tissue samples from one cat (F159), we also attempted to amplify the htrA locus ( 36 ). To detect Bartonella , primers of the pap31 locus of the hemin binding protein, PAPn1 and PAPn2 were used (Additional file 1: Table S1 ) ( 37 ). All PCRs used the Failsafe™ PCR kits (Lucigen, Wisconsin, USA). Resulting PCR fragments were visualized on 1% agarose gel, treated with ExoSapIT (Applied Biosystems, Waltham, MA) and sent to Eton Biosciences (San Diego, CA) for Sanger dideoxy sequencing. The DNA sequences were analyzed using MEGA7 ( 38 ) and Geneious Prime (Biomatters Inc. Boston, MA) and queried against the NCBI Genbank database. We report results of sequences with a match of 97% identity and an E value ≤ 0 to a named species. A sample was called positive only when we retrieved a sequence that met these criteria for at least one locus. Sequences that did not meet these criteria but indicated Rickettsia or Bartonella were assigned genus-level identification. Sequences have been deposited into GenBank ( gltA : PP856019-PP856043; ompB : PP935775-PP935802; pap31 : PP943066-PP943101; PP944938-PP944992; PP947812-PP947814). Flea infestation prevalence of cats was defined as the proportion of sampled cats that harbored at least one flea. Flea burden was defined as the number of fleas per infested host. To test for associations between cat demographic factors (sex, geographic origin) and season with the outcomes of flea burden and/or flea infection, we used Chi-square, Fisher’s exact, Wilcoxon Rank Sum, and Kruskal-Wallis tests using the program R. Results Cats from Hidalgo and Cameron counties in Texas were sampled from January-June 2017. Cats were from 14 different urban locations, with most coming from the cities of McAllen (66), Pharr ( 28 ), Edinburg ( 20 ), and San Juan ( 18 ) (Fig. 1 ; Table 1 ). Of the 167 cats in the study, 87 were female and 80 were male. Most cats were sampled in the winter months (72%), fewer in spring (17%) and summer (11%) (Table 1 ). Table 1 Infection of fleas removed from stray cats and cat blood with Rickettsia and Bartonella across 14 locations of the Rio Grande Valley of south Texas, 2017. Cat blood tested for Rickettsia had one positive (F112). No. of Cats Flea infested cats (%) No. flea pools tested Candidatus Rickettsia senegalensis positive flea pools (%) Rickettsia typhi positive flea pools (%) Bartonella henselae positive flea pools (%) Bartonella clarridgeiae positive flea pools (%) Bartonella henselae positive cat blood (% cats) Bartonella clarridgeiae positive cat blood (% cats) Location Alamo 10 8(80) 8 3( 38 ) 1( 13 ) 6(75) 5( 50 ) Alton 3 3(100) 3 2(67) 2(67) 1( 33 ) Donna 4 4(100) 4 1( 25 ) Edinburg 20 14(70) 14 4( 29 ) 7( 50 ) 3( 15 ) Elsa 2 2(100) 2 2(100) 1( 50 ) Harlingen 1 1(100) 1 1(100) McAllen 66 55(83) 55 10( 18 ) 4( 7 ) 16( 29 ) 1( 2 ) 5( 8 ) Mercedes 2 1( 50 ) 1 1( 50 ) Mission 5 5(100) 5 1( 20 ) 3(60) 1( 20 ) Monte Alto 1 1(100) 1 Palmview 5 5(100) 5 1( 20 ) 2( 40 ) Pharr 28 22(79) 22 2( 9 ) 1( 5 ) 14(64) 2( 9 ) 10( 36 ) San Juan 18 16(89) 17 4( 24 ) 5( 29 ) 9( 50 ) 1( 6 ) Weslaco 2 2(100) 2 1( 50 ) 1( 50 ) Season Spring 28 25(89) 25 1( 4 ) 8( 32 ) 10( 36 ) Summer 19 13(68) 13 2( 15 ) 1( 8 ) 6( 46 ) 1( 5 ) Winter 120 101(84) 102 25( 25 ) 5( 5 ) 45( 44 ) 3( 3 ) 26( 22 ) 1( 1 ) Sex Female 87 67(77) 67 8( 12 ) 3( 4 ) 32( 48 ) 1( 1 ) 19( 22 ) 1( 1 ) Male 80 73(91) 73 20( 27 ) 3( 4 ) 27( 37 ) 2( 3 ) 18( 23 ) Total 167 139(83) 140 28( 20 ) 6( 4 ) 59( 42 ) 3( 2 ) 37( 22 ) 1( 1 ) A total of 721 fleas were collected from 139 cats resulting in an infestation prevalence of 83% (Table 1 ). Flea burden ranged from 1 to 41. An average of 4.3 fleas per cat (5.2 fleas per infested cat) were found. Significantly more male than female cats harbored fleas, with 91% and 76% of male and female cats infested, respectively (Wilcoxon Rank Sum Test, W = 2607, p-value = 0.004). Infestation prevalence was lowest in the summer months, with fleas apparent on 101 of 120 (84%) of cats sampled in winter, 25 of 28 (89%) in spring and 13 of 19 (68%) in summer (Kruskal-Wallis test, H = 12.43, dƒ = 2, p-value = 0.002). Most fleas (n = 718) were C. felis (cat flea), with two Echidnophaga gallinacea (sticktight flea) and one single Pulex sp . Most infested cats harbored only a single flea species, but cat F63 harbored a single E. gallinacea and four C. felis . Of 140 flea pools tested for Rickettsia , 34 (24%) were positive of which 28 (20% of all flea pools) were positive for Candidatus Rickettsia senegalensis and 6 (4% of all flea pools) were positive for R. typhi . Rickettsia -positive fleas were collected from 9 of the 14 locations, including McAllen ( 14 ), four each from Alamo, Edinburg and San Juan, three from Pharr, two from Alton and one each from Donna, Mission and Palmview. Rickettsia infection did not show association with location (Chi-square test, χ 2 = 10.132, dƒ = 13, p = 0.683). Rickettsia infection was positively associated with flea burden (Wilcoxon Rank Sum Test, W = 968, p-value = 1.968e-07). Further, Rickettsia infection prevalence in fleas was greatest in winter (30 of 34), whereas only three and one infected flea pools were found in summer and spring, respectively (Chi-square test, χ 2 = 6.607, dƒ = 2, p = 0.0368). A total of 11 of 67 flea pools (16%) from female cats and 23 of 73 flea pools (32%) from male cats were positive for Rickettsia (Chi-square test, χ 2 = 5.885, dƒ = 1, p = 0.0153). Of the 6 R. typhi -positive flea pools, four were from McAllen and one each were from Pharr and Alamo (Table 1 ). Additionally, non-target amplification of Bartonella occurred with two Rickettsia assays, as one flea pool (F72) amplified using the RrCS gltA primers showed a B. clarridgeiae sequence; three flea pools amplified using the RpCS gltA primers showed B. henselae sequences; and five flea pools amplified using the RpCS gltA primers resulted in genus-level identifications to Bartonella . All the pools that showed Bartonella from these two assays were also detected by the Bartonella -specific pap31 PCR (Additional file 1: Table S1 ). PCR amplification of the Bartonella pap 31 locus detected was positive in 90 flea pools (64% of all pools), of which 59 (42% of all pools) were confirmed B. henselae , 3 (2% of all pools) were B. clarridgeiae and another 28 were identified to the genus level as Bartonella . Bartonella status did not correlate with the location of the host (Chi-square test, χ 2 = 11.135, dƒ = 13, p = 0.6). There were no statistical differences in seasonal distribution of Bartonella positivity, with 55% winter, 61% spring and 36% summer pools of each season testing positive (χ 2 = 2.758;, dƒ = 2, p = 0.252). A total of 48 pools from female cats and 42 from male cats carried Bartonella (Chi-square test, χ 2 = 0.012238, dƒ = 1, p-value = 0.912) (Table 1 ). Lastly, coinfections of flea pools with both Rickettsia and Bartonella occurred more commonly than expected by chance (Chi-square test, χ2 = 5.858, dƒ = 1, p value = 0.0155), with 25 flea pools positive for both Rickettsia and Bartonella . This included five of the six R. typhi -positive flea pools which all caried Bartonella of which 4 were B. henselae . Blood clot and heart tissue DNA from 24 cats was evaluated using the Rickettsial ompB locus; these included all six cats that harbored R . typhi -positive fleas, 11 cats that harbored fleas with Candidatus Rickettsia senegalensis and seven cats with Rickettsia -negative fleas. An expanded set of other tissues was evaluated from one cat with R . typhi -positive fleas (F159) and three with negative fleas (F154, F156 and F157). Although PCR products using ompB , RrCS and in one case htrA were observed in some of these samples, only one instance (F112 blood clot) yielded sequence matching Candidatus R. senegalensis. Others did not match Rickettsia or yielded poor quality sequences(Additional file 1: Table S2), and thus the cats were considered negative. We examined DNA extracted from blood clots of all 167 cats for Bartonella using PCR. Thirty-nine samples on the pap31 PCR were positive – 37 (22% of cats) carried Bartonella henselae , one (0.6%) carried Bartonella clarridgeiae and one was only identifiable to genus. When Bartonella in cats and fleas were compared, 30 flea pools and matching cats showed carriage of Bartonella (χ 2 = 8.595, dƒ = 1, p = 0.0034). The proportion of cats carrying Bartonella was highest in cats from San Juan (56%). Seasonally, 28 of 120 (23%) cats sampled in winter showed Bartonella (26 B. henselae , one B. clarridgeiae , one Bartonella spp.) while 10 of 28 (36%) in spring (all B. henselae ) and one of 19 (5%) in summer (one B. henselae ). The B. henselae infection prevalence was 24% in both female (21/87) and male (19/80) cats. Discussion This study strengthens evidence that south Texas harbors robust enzootic, cat flea-driven cycles of transmission of R. typhi , associated with the emergence of murine typhus in humans in the area. Further, we provide new evidence of enzootic cycles of B. henselae transmission. In a recent study from Galveston, TX ( 21 ), R. typhi was detected in fleas from one of 24 feral cats (4.1%), remarkably similar to 4.3% of 140 flea pools we report here. The Galveston study also detected Candidatus R. senegalensis as we did, in addition to two species that were not apparent in our study, R. asemboensis and R. felis . In contrast, R. typhi was not detected in a survey of fleas from 283 feral/stray cats from southeastern Georgia, USA, in which 16.5% of flea pools were positive for the 17-kDa protein antigen gene of Rickettsia spp., none of which were positive for ompB of R. typhi ( 39 ). Despite the relatively high infection prevalence of Bartonella we detected in both vectors (64% of flea pools) and hosts (23% of cats), relatively little ecological work has been done on this agent in the southern USA. In southeastern Georgia, USA, 35.2% of flea pools from feral/stray cats were infected with Bartonella ( 39 ). In a study of cats from catteries across North America, a seroprevalence of 35.8% was recorded, in which flea infestation was the most important risk factor for high B. henselae seroprevalence ( 27 ). Lower Bartonella infection prevalence among fleas removed from cats have been reported in studies from Canada (4.3%), Chile (7.5%), Ethiopia (6%) and Greece (13.5%) ( 40 – 43 ). As expected, we found cats with Bartonella -positive blood commonly harbored infected fleas, yet there were also several infected flea pools removed from cats whose blood and tissues tested negative, suggesting that those fleas may have picked up the infection from a prior bloodfeeding event on a different host. Similarly, a study of cats in Brazil found Bartonella DNA in 47.8% of the cat blood samples and 18.3% of C. felis fleas, in which cats infested by positive ectoparasites showed approximately twice the odds of being infected ( 24 ). Two cats (F31 and F33) simultaneously harbored fleas infested with both R. typhi and B. henselae , potentially serving as companion animal sentinels of multiple regional zoonotic threats to humans. Further, this exact sample set of cats has also been examined in previous studies for additional zoonoses, including Trypanosoma cruzi , the causal agent of Chagas disease (19/167 cats, 11.4% prevalence) and Dirofilaria immitis (canine heartworm) which occasionally causes pulmonary disease in humans (22/122 cats, 18% prevalence) ( 29 , 44 ). In comparing the results across studies, two cats (F96 and F141) were infected with Bartonella , T. cruzi and D. immitis ; five (F96, F101, F133, F141 and F142) with Bartonella and T. cruzi ; and eight with Bartonella and D. immitis . One T. cruzi -positive cats harbored R. typhi -positive fleas (F35), and two T. cruzi -positive cats harbored Candidatus R. senegalensis-positive fleas (F26 and F113). While an older study suggested pet ownership is not a direct risk factor for human exposure to R. typhi based on Texas data ( 45 ) owning a dog was found to be a risk factor for murine typhus seropositivity in a cross-sectional study from India ( 46 ). While feral/stray cats that remain outdoors may pose little risk of zoonoses transmission from flea importation to the house, those cats that enter homes and those that interact with domestic animals may facilitate ‘zoonoses in the bedroom’ by introducing new disease threats to the domestic setting ( 47 ). Cases of murine typhus have been steadily increasing in the United States over the past three decades with 348 cases from 1990-99, 994 from 2000-09, a stark 3159 from 2010–2018. A total of 6700 cases were reported in Texas between 2008-23 ( https://www.dshs.texas.gov ). Within Texas, the Rio Grande Valley appears to be a hotspot for local cases where Hidalgo County- the location of this current study- reported 1383 cases between 2008 to 2023 ( https://www.dshs.texas.gov ). In an 18-year period from 1998 to 2016, 213 pediatric patients were admitted to a hospital in south Texas with a confirmed diagnosis of murine typhus ( 48 ). Nationally, cat scratch disease is diagnosed for 4.5–9.3 patients per 100,000 people, highest in the southern United States, in children aged 5–9, and with hospitalization higher among males 50–64 years old ( 12 , 49 ). In a survey across 9 years of nearly 40 million medical insurance enrollees, 0.03% were diagnosed with cat scratch disease with nearly one-fifth (19.7%) of cases from the West South Central US census region that includes Texas, second to 26.3% in the South Atlantic US census region ( 12 ). In a similar analysis, over a ten-year period, over half the diagnosis of cat scratch disease was from a southern state in the US ( 16 ). Although it is established that Bartonella infections are life threatening for immunocompromised individuals, neurobartonellosis in immunocompetent individuals is incompletely understood ( 17 ) and the public health burden of bartonellosis is unknown. Cats are deemed to be adapted to B. henselae but non-adapted species of Bartonella and occasional variants of B. henselae can cause cardiac disease in cats ( 19 , 20 ). New Rickettsia and Bartonella species and novel vector-host associations continue to be described ( 50 – 52 ), underscoring the need for vector and host surveillance to provide an ecological basis for public and veterinary health risk assessment. Controlling fleas and infections in domestic cats may help to alleviate spillover transmission to humans. Declarations Ethics approval and consent to participate The Texas A&M. University Institutional Animal Care and Use Committee (IACUC) issued a letter of exemption on December 16, 2016, as samples were collected exclusively from cats that were euthanized for non-study reasons. Consent for publication Not Applicable Competing interests The authors declare that they have no competing interests Funding Texas A&M AgriLife Research provided funding. Author Contribution SB: conceptualization; data curation; formal analysis; investigation; writing- original draft; writing- review & editing.IZ: conceptualization; investigation; writing- review & editing.AK: investigation; methodology; writing- review & editing.SH: conceptualization; funding acquisition; investigation; project administration; resources; writing- review & editing. Acknowledgement We thank Lisa Auckland and Keswick Killets for assistance in the laboratory and Dr. Jose Juarez-Valdez for the map for Figure 1. Graphical abstract was created in BioRender. Balasubramanian, S. (2026) https://BioRender.com/kssvz5w Data Availability The datasets generated and/or analysed during the current study are available in the OAKTrust Digital Repository [https://oaktrust.library.tamu.edu/home](https:/oaktrust.library.tamu.edu/home) . References Fang R, Blanton LS, Walker DH. Rickettsiae as emerging infectious agents. Clin Lab Med. 2017;37(2):383–400. Kyle G. Rodino. Rickettsioses in the United States | Elsevier Enhanced Reader. Clin Microbiol Newsl. 2019;41(13):113–9. Azad AF, Radulovic S, Higgins JA, Noden BH, Troyer JM. Flea-borne rickettsioses: ecologic considerations. Emerg Infect Dis. 1997;3(3):319–27. Pérez-Osorio CE, Zavala-Velázquez JE, León JJA, Zavala-Castro JE. Rickettsia felis as Emergent Global Threat for Humans. Emerg Infect Dis. 2008;14(7):1019–23. Anstead GM. History, rats, fleas, and opossums. II. The decline and resurgence of flea-borne typhus in the United States, 1945–2019. Trop Med Infect Dis. 2020;6(1):2. Civen R, Ngo V. Murine typhus: An unrecognized suburban vectorborne disease. Clin Infect Dis. 2008;46(6):913–8. Caravedo Martinez MA, Ramírez-Hernández A, Blanton LS. Manifestations and management of flea-borne Rickettsioses. Res Rep Trop Med. 2021;Volume 12:1–14. Juhasz NB, Wilson JM, Haney KN, Clark MH, Davenport AC, Breitschwerdt EB, et al. Rickettsia typhi infection in a clinically-ill dog from Houston, Texas. Vet Parasitol Reg Stud Rep. 2022;35:100781. Sorvillo FJ, Gondo B, Emmons R, Ryan P, Waterman SH, Tilzer A, et al. A suburban focus of endemic typhus in Los Angeles county: association with seropositive domestic cats and opossums. Am J Trop Med Hyg. 1993;48(2):269–73. Gracia MJ, Marcén JM, Pinal R, Calvete C, Rodes D. Prevalence of Rickettsia and Bartonella species in Spanish cats and their fleas. J Vector Ecol. 2015;40(2):233–9. Chomel BB, Boulouis HJ, Breitschwerdt EB, Kasten RW, Vayssier-Taussat M, Birtles RJ, et al. Ecological fitness and strategies of adaptation of Bartonella species to their hosts and vectors. Vet Res. 2009;40(2):29. Nelson CA, Saha S, Mead PS. Cat-Scratch disease in the United States, 2005–2013. Emerg Infect Dis. 2016;22(10):1741–6. Kordick DL, Papich MG, Breitschwerdt EB. Efficacy of enrofloxacin or doxycycline for treatment of Bartonella henselae or Bartonella clarridgeiae infection in cats. Antimicrob Agents Chemother. 1997;41(11):2448–55. Jacomo V, Kelly PJ, Raoult D. Natural History of Bartonella Infections (an Exception to Koch’s Postulate). Clin Vaccine Immunol. 2002;9(1):8–18. Jurja S, Stroe AZ, Pundiche MB, Docu Axelerad S, Mateescu G, Micu AO, et al. The clinical profile of cat-scratch disease’s neuro-ophthalmological effects. Brain Sci. 2022;12(2):217. Nawrocki CC, Max RJ, Marzec NS, Nelson CA. Atypical manifestations of cat-scratch disease, United States, 2005–2014 - Volume 26, Number 7—July 2020 - Emerging Infectious Diseases journal - CDC. Emerg Infect Dis [Internet]. 2020 Jul [cited 2022 Sep 20];26(7). Available from: https://wwwnc.cdc.gov/eid/article/26/7/20-0034_article Delaney S, Robveille C, Maggi RG, Lashnits E, Kingston E, Liedig C, et al. Bartonella species bacteremia in association with adult psychosis. Front Psychiatry. 2024;15:1388442. Rolain JM, La Scola B, Liang Z, Davoust B, Raoult D. Immunofluorescent detection of intraerythrocytic Bartonella henselae in naturally infected cats. J Clin Microbiol. 2001;39(8):2978–80. Kordick DL, Brown TT, Shin K, Breitschwerdt EB. Clinical and pathologic evaluation of chronic Bartonella henselae or Bartonella clarridgeiae infection in cats. J Clin Microbiol. 1999;37(5):1536–47. Breitschwerdt EB, Lappin MR. Feline bartonellosis: we’re just scratching the surface. J Feline Med Surg. 2012;14(9):609–10. Blanton LS, Vohra RF, Fistein L, Quade B, Walker DH, Bouyer DH. Rickettsiae within the fleas of feral cats in Galveston, Texas. Vector-Borne Zoonotic Dis. 2019;19(9):647–51. Huang HHH, Power RI, Mathews KO, Ma GC, Bosward KL, Šlapeta J. Cat fleas (Ctenocephalides felis clade ‘Sydney’) are dominant fleas on dogs and cats in New South Wales, Australia: Presence of flea-borne Rickettsia felis , Bartonella spp. but absence of Coxiella burnetii DNA. Curr Res Parasitol Vector-Borne Dis. 2021;1:100045. Liodaki M, Spanakos G, Samarkos M, Daikos GL, Christopoulou V, Piperaki ET. Molecular screening of cat and dog ectoparasites for the presence of Bartonella spp. in Attica, Greece. Acta Vet Hung. 2022; Raimundo JM, Guimarães A, Amaro GM, Silva AT da, Rodrigues CJBC, Santos HA, et al. Prevalence of Bartonella species in shelter cats and their ectoparasites in southeastern Brazil. Rev Bras Parasitol Veterinária. 2022;31(1):e014221. Razgūnaitė M, Lipatova I, Paulauskas A, Karvelienė B, Riškevičienė V, Radzijevskaja J. Bartonella infections in cats and cat fleas in Lithuania. Pathogens. 2021;10(9):1209. Bitam I, Dittmar K, Parola P, Whiting MF, Raoult D. Fleas and flea-borne diseases. Int J Infect Dis. 2010;14(8):e667–76. Foley JE, Chomel B, Kikuchi Y, Yamamoto K, Pedersen NC. Seroprevalence of Bartonella henselae in cattery cats: Association with cattery hygiene and flea infestation. Vet Q. 1998;20(1):1–5. Iannino. Fleas of dog and cat: species, biology and flea-borne diseases. Vet Ital. 2017;53(4):273–5. Zecca IB, Hodo CL, Slack S, Auckland L, Rodgers S, Killets KC, et al. Prevalence of Trypanosoma cruzi infection and associated histologic findings in domestic cats (Felis catus). Vet Parasitol. 2020;278:109014. Lawrence AL, Webb CE, Clark NJ, Halajian A, Mihalca AD, Miret J, et al. Out-of-Africa, human-mediated dispersal of the common cat flea, Ctenocephalides felis : The hitchhiker’s guide to world domination. Int J Parasitol. 2019;49(5):321–36. Centers for Disease Control. Pictorial keys to arthropods, reptiles, birds, and mammals of public health significance [Internet]. 1966 [cited 2025 Dec 15]. Available from: https://stacks.cdc.gov Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol. 1994;3(5):294–9. Roux V, Raoult D. Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol. 2000;50(4):1449–55. Williamson PC, Billingsley PM, Teltow GJ, Seals JP, Turnbough MA, Atkinson SF. Borrelia , Ehrlichia , and Rickettsia spp. in ticks removed from persons, Texas, USA. Emerg Infect Dis. 2010;16(3):441–6. Regenery RL, Spruill CL, Plikaytis BD. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes [Internet]. 1991 [cited 2022 Mar 1]. Available from: https://journals.asm.org/doi/epdf/ 10.1128/jb.173.5.1576-1589.1991 Webb L, Mitchell C, Malloy DC, Dasch GA, Azad AF. Detection of murine typhus infection in fleas by using the polymerase chain reaction [Internet]. 1990 [cited 2022 Mar 1]. Available from: https://journals.asm.org/doi/epdf/ 10.1128/jcm.28.3.530-534.1990 Zeaiter Z, Fournier PE, Ogata H, Raoult D. Phylogenetic classification of Bartonella species by comparing groEL sequences. Int J Syst Evol Microbiol. 2002;52(1):165–71. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33(7):1870–4. Brown LD, Maness R, Greer K. Detection of Bartonella spp. and Rickettsia spp. in cat fleas ( Ctenocephalides felis ) collected from free-roaming domestic cats in southeastern Georgia, USA. Vet Parasitol Reg Stud Rep. 2022;100743. Liodaki M, Spanakos G, Samarkos M, Daikos GL, Christopoulou V, Piperaki ET. Molecular screening of cat and dog ectoparasites for the presence of Bartonella spp. in Attica, Greece. Acta Vet Hung. 2022;70(1):9–14. Müller A, Rodríguez E, Walker R, Bittencourt P, Pérez-Macchi S, Gonçalves LR, et al. Occurrence and genetic diversity of Bartonella spp. (Rhizobiales: Bartonellaceae) and Rickettsia spp. (Rickettsiales: Rickettsiaceae) in cat fleas (Siphonaptera: Pulicidae) From Chile. J Med Entomol. 2018;55(6):1627–32. Kumsa B, Parola P, Raoult D, Socolovschi C. Molecular Detection of Rickettsia felis and Bartonella henselae in dog and cat fleas in central Oromia, Ethiopia. Am J Trop Med Hyg. 2014;90(3):457–62. Kamrani A, Parreira VR, Greenwood J, Prescott JF. The prevalence of Bartonella, hemoplasma, and Rickettsia felis infections in domestic cats and in cat fleas in Ontario. Can J Vet Res. 2008;72(5):411–9. Mosley IA, Zecca IB, Tyagi N, Harvey TV, Hamer SA, Verocai GG. Occurrence of Dirofilaria immitis infection in shelter cats in the lower Rio Grande Valley region in South Texas, United States, using integrated diagnostic approaches. Vet Parasitol Reg Stud Rep. 2023;41:100871. Wiggers RJ, Stewart RS. Ownership of cats or dogs does not increase exposure to Rickettsia typhi . Tex Med. 2002;98(6):56–7. Devamani CS, Schmidt WP, Ariyoshi K, Anitha A, Kalaimani S, Prakash JAJ. Risk factors for scrub typhus, murine typhus, and spotted fever seropositivity in urban areas, rural plains, and peri-forest hill villages in south India: a cross-sectional study. Am J Trop Med Hyg. 2020;103(1):238–48. Chomel BB, Sun B. Zoonoses in the bedroom. Emerg Infect Dis. 2011;17(2):167–72. Howard A, Fergie J. Murine Typhus in South Texas Children: An 18-year Review. Pediatr Infect Dis J. 2018;37(11):1071–6. Nelson CA, Moore AR, Perea AE, Mead PS. Cat scratch disease: U.S. clinicians’ experience and knowledge. Zoonoses Public Health. 2018;65(1):67–73. do Amaral RB, Cardozo MV, Varani A de M, Furquim MEC, Dias CM, Assis WO de, et al. First report of Bartonella spp. in marsupials from Brazil, with a description of Bartonella harrusi sp. nov. and a new proposal for the taxonomic reclassification of species of the genus Bartonella . Microorganisms. 2022;10(8):1609. Medkour H, Lo CI, Anani H, Fenollar F, Mediannikov O. Bartonella massiliensis sp. nov., a new bacterial species isolated from an Ornithodoros sonrai tick from Senegal. New Microbes New Infect. 2019;32:100596. Laroche M, Berenger JM, Mediannikov O, Raoult D, Parola P. Detection of a Potential New Bartonella Species “ Candidatus Bartonella rondoniensis” in Human Biting Kissing Bugs (Reduviidae; Triatominae). Gürtler RE, editor. PLoS Negl Trop Dis. 2017;11(1):e0005297. Additional Declarations No competing interests reported. Supplementary Files AdditionalFile1.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 04 Feb, 2026 Editor assigned by journal 03 Feb, 2026 Submission checks completed at journal 03 Feb, 2026 First submitted to journal 28 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8726915","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":585056571,"identity":"ae06dad9-596a-4283-a211-8a029fe8e5c4","order_by":0,"name":"Sujata Balasubramanian","email":"","orcid":"","institution":"Texas A\u0026M University","correspondingAuthor":false,"prefix":"","firstName":"Sujata","middleName":"","lastName":"Balasubramanian","suffix":""},{"id":585056572,"identity":"802b792f-406b-48b6-9025-230838676e8d","order_by":1,"name":"Italo Zecca","email":"","orcid":"","institution":"Texas A\u0026M University","correspondingAuthor":false,"prefix":"","firstName":"Italo","middleName":"","lastName":"Zecca","suffix":""},{"id":585056573,"identity":"0dc9e5b8-366f-491e-880f-4c2ec121af56","order_by":2,"name":"Allyson Koger","email":"","orcid":"","institution":"Texas A\u0026M University","correspondingAuthor":false,"prefix":"","firstName":"Allyson","middleName":"","lastName":"Koger","suffix":""},{"id":585056574,"identity":"33d6fda4-c258-48d8-8979-4222e90dbd0d","order_by":3,"name":"Sarah Hamer","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0ElEQVRIiWNgGAWjYDACCTBpgyyUQJSWNBDB2ECKlsMkaJGf3fx0w8c95+UNrh1gf/Az5zADP3uOAV4tBneOmd2c8ey24YbbCYyNvdsOM0j2vCGgRSLB7DbPgdsJkrMTGBt4gVoMbhCwRX5G+rfbfw6cA2tp/AvUYk9IC8ONHLPbDAcOJPBLJzA2g22RIOSXGzllN3sOJBv2Syc2zpbdls4jceZZASGHbbvx44CdPJt08oGPb7dZy/G3J2/A7zAEgEQLD7HKR8EoGAWjYBTgAQCCeUuZOi7VVAAAAABJRU5ErkJggg==","orcid":"","institution":"Texas A\u0026M University","correspondingAuthor":true,"prefix":"","firstName":"Sarah","middleName":"","lastName":"Hamer","suffix":""}],"badges":[],"createdAt":"2026-01-29 04:24:45","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8726915/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8726915/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101840743,"identity":"a7d0cc88-441b-44d5-9776-74f10d5b2412","added_by":"auto","created_at":"2026-02-04 08:30:22","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1026725,"visible":true,"origin":"","legend":"\u003cp\u003e(Figure will be submitted in a separate file. Only legend will go here) Map showing the locations of origin within Hidalgo County, Texas from where the cats in the study were surrendered. Map was generated using Quantum GIS (QGIS 3.30) using freely available administrative boundaries and map data from OpenStreetMap for satellite imagery. Open Street Map. Planet OSM [Internet]. [cited 2025 Dec 15]. Available from: https://planet.osm.org/\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8726915/v1/1bf607fc5c59c89c4114fb65.png"},{"id":101881795,"identity":"cf5a3448-6963-497f-bba0-b24745258fe0","added_by":"auto","created_at":"2026-02-04 15:16:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1716158,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8726915/v1/a344c3aa-7b7a-4d25-83a9-2ef69ca17d77.pdf"},{"id":101840744,"identity":"c1a5ecef-ca40-472e-9acd-abf85cd5fa28","added_by":"auto","created_at":"2026-02-04 08:30:22","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":24733,"visible":true,"origin":"","legend":"","description":"","filename":"AdditionalFile1.docx","url":"https://assets-eu.researchsquare.com/files/rs-8726915/v1/3ef84f43603e009c7383d9cf.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Rickettsia typhi, Bartonella henselae and related zoonotic agents in fleas from domestic cats (Felis catus) from the Rio Grande Valley, Texas ","fulltext":[{"header":"Background","content":"\u003cp\u003eFlea-borne diseases continue to emerge in human populations worldwide, with increasing attention to pathogens in the genera \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e. At least 17 species of \u003cem\u003eRickettsia\u003c/em\u003e cause disease in humans, many resulting in spotted fever or typhus fever (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). \u003cem\u003eRickettsia typhi\u003c/em\u003e \u0026ndash; the causative agent of murine typhus or flea-borne typhus - is an emergent global threat for humans (\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). When typhus was initially identified in the 1920s, rats were the reservoirs, and the rat flea (\u003cem\u003eXenopsylla cheopis)\u003c/em\u003e was the recognized vector. \u003cem\u003eR. typhi\u003c/em\u003e is now also transmitted by \u003cem\u003eCtenocephalides felis\u003c/em\u003e (cat flea) and other flea species (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e), with opossums (\u003cem\u003eDidelphis virginiana\u003c/em\u003e) and cats (\u003cem\u003eFelis catus\u003c/em\u003e) as reservoirs (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). \u003cem\u003eR. typhi\u003c/em\u003e is endemic to south Texas, California, and Hawaii in the US. Infection with \u003cem\u003eR. typhi\u003c/em\u003e may cause fever, headache, chills, rash, gastrointestinal or renal symptoms with potential for hospitalization, though mortality is rare and treatment is accomplished with antibiotics (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Clinical impacts on infected pets are not well studied, although infected cats have been reported from many human disease hotspots, and one case study identified \u003cem\u003eR. typhi\u003c/em\u003e in a clinically ill dog in Texas (\u003cspan additionalcitationids=\"CR9\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMultiple species of \u003cem\u003eBartonella\u003c/em\u003e are recognized as emerging threats to public health (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). \u003cem\u003eBartonella henselae\u003c/em\u003e is the main causal agent of cat scratch disease is but \u003cem\u003eB. clarridgeiae\u003c/em\u003e is also considered a causal agent (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). In humans, infection with \u003cem\u003eBartonella\u003c/em\u003e has different outcomes for immunocompetent and immunocompromised human hosts (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). Humans acquire bartonellosis from bites or scratches from cats, more frequently diagnosed in children. Cat-scratch disease in humans progresses acutely from skin lesion to fever and lymphadenopathy. Neurological complications, often ocular, have been observed with \u003cem\u003eBartonella\u003c/em\u003e spp infections (\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). In cats, \u003cem\u003eB. henselae\u003c/em\u003e is maintained in an intra-erythrocytic state and typically results in asymptomatic bacteremia (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e), although infected cats may develop cardiovascular issues including lymphoplasmacytic myocarditis among other ailments affecting the lymph nodes, spleen, liver, and kidneys; sometimes fatal (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe cat flea (\u003cem\u003eC. felis\u003c/em\u003e) is a predominant vector for \u003cem\u003eR. typhi\u003c/em\u003e, \u003cem\u003eB. hensela\u003c/em\u003ee and related species. This flea has a worldwide distribution and is an abundant and frequent ectoparasite on domestic dogs and cats. Across broad geographic ranges, \u003cem\u003eC. felis\u003c/em\u003e is the most common flea species found on domestic and feral cats, often among other species including the sticktight flea (\u003cem\u003eEchidnophaga gallinacea\u003c/em\u003e) and human flea (\u003cem\u003ePulex irritans\u003c/em\u003e) (\u003cspan additionalcitationids=\"CR22 CR23 CR24\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e). In addition to \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e spp. pathogens, \u003cem\u003eC. felis\u003c/em\u003e may transmit or carry \u003cem\u003eYersinia pestis\u003c/em\u003e (plague), \u003cem\u003eDipylidium caninum\u003c/em\u003e (tapeworm), \u003cem\u003eHymenolepis diminuta\u003c/em\u003e (rat tapeworm), \u003cem\u003eDipetalonema\u003c/em\u003e (\u003cem\u003eAcanthocheilonema\u003c/em\u003e) \u003cem\u003ereconditum\u003c/em\u003e and hemoplasmas (\u003cspan additionalcitationids=\"CR27\" citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e), and may directly impact hosts through flea allergic dermatitis.\u003c/p\u003e \u003cp\u003eConsidering the emergent threat of \u003cem\u003eR. typhi\u003c/em\u003e and \u003cem\u003eB. henselae\u003c/em\u003e in the U.S., our work aims to describe the prevalence of both genera among fleas and their cat hosts in the Rio Grande Valley in south Texas.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eThe Rio Grande Valley of south Texas is a region with a subtropical climate that supports many vector species and tropical infections. We partnered with a large shelter in Hidalgo County that intakes an estimated 9,000 cats per year. Cats in our study included 167 predominantly stray cats from 14 surrounding rural and urban zones of the Rio Grande Valley, and were the subject of a prior study of \u003cem\u003eTrypanosoma cruzi\u003c/em\u003e and feline Chagas disease (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e). These cats were sampled across winter (01 Jan-19 Mar), spring (20 Mar\u0026thinsp;\u0026minus;\u0026thinsp;20 June) and summer (21 June \u0026ndash; 19 Sept) of 2017. At the time of euthanasia for non-study reasons, the sex, and geographic origin of cats was recorded. Cats were combed using flea combs across the whole body for approximately three minutes and all recovered fleas were preserved in 70% ethanol.\u003c/p\u003e \u003cp\u003eAs described previously (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e), blood was collected from all cats, spun and separated in the laboratory with DNA extracted from the blood clot available for the current study. Additionally, DNA extracted from heart was available from all cats and other tissues were also available from cats sampled in spring and summer.\u003c/p\u003e \u003cp\u003eFleas were identified using the key from Centers for Disease Control and Prevention with pictorial details from Lawrence et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e). When \u0026le;5 conspecific fleas were collected from an individual cat, they were pooled for DNA extraction. When \u0026gt;\u0026thinsp;5 conspecific fleas were present, 5 were randomly selected to create a pool. When \u0026gt;\u0026thinsp;1 flea species was present, a pool was made for each species. Representative fleas have been deposited into Texas A\u0026amp;M University Insect Collection (TAMUIC).\u003c/p\u003e \u003cp\u003eThe Omega EZNA (Omega Bio-tek, Norcross, GA) kit was used to extract DNA from fleas. Fleas were first crushed into lysis buffer using single-use pestles. To confirm the flea species identification, representatives of each species were subjected to a molecular identification process using PCR targeting the insect cytochrome oxidase I locus with primers LCO1490 and HC02198 (Additional file 1: Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). PCR products were sequenced, and chromatograms were analyzed as described below. Representative flea sequences were deposited to Genbank (PP545303-PP545306).\u003c/p\u003e \u003cp\u003eThree different PCRs were used to identify \u003cem\u003eRickettsia\u003c/em\u003e (Additional file 1: Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) from flea pools and selected cat blood clots and tissues. Outer membrane protein (\u003cem\u003erOmpB\u003c/em\u003e) was amplified using primers 120\u0026ndash;2788 and 120\u0026ndash;3599; this assay can detect a wide range of Rickettsial species (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e). Two sets of primers were used to amplify the \u003cem\u003egltA\u003c/em\u003e (citrate synthase) gene: RrCS to detect spotted fever group \u003cem\u003eRickettsia\u003c/em\u003e using primers RrCS 372 and RrCS 989 (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e) and RpCS to detect the typhus group \u003cem\u003eRickettsia\u003c/em\u003e using primers RpCS 877 and 1258n (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). For the blood and tissue samples from one cat (F159), we also attempted to amplify the \u003cem\u003ehtrA\u003c/em\u003e locus (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo detect \u003cem\u003eBartonella\u003c/em\u003e, primers of the \u003cem\u003epap31\u003c/em\u003e locus of the hemin binding protein, PAPn1 and PAPn2 were used (Additional file 1: Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). All PCRs used the Failsafe\u0026trade; PCR kits (Lucigen, Wisconsin, USA). Resulting PCR fragments were visualized on 1% agarose gel, treated with ExoSapIT (Applied Biosystems, Waltham, MA) and sent to Eton Biosciences (San Diego, CA) for Sanger dideoxy sequencing.\u003c/p\u003e \u003cp\u003eThe DNA sequences were analyzed using MEGA7 (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e) and Geneious Prime (Biomatters Inc. Boston, MA) and queried against the NCBI Genbank database. We report results of sequences with a match of 97% identity and an E value\u0026thinsp;\u0026le;\u0026thinsp;0 to a named species. A sample was called positive only when we retrieved a sequence that met these criteria for at least one locus. Sequences that did not meet these criteria but indicated \u003cem\u003eRickettsia\u003c/em\u003e or \u003cem\u003eBartonella\u003c/em\u003e were assigned genus-level identification. Sequences have been deposited into GenBank (\u003cem\u003egltA\u003c/em\u003e: PP856019-PP856043; \u003cem\u003eompB\u003c/em\u003e: PP935775-PP935802; \u003cem\u003epap31\u003c/em\u003e: PP943066-PP943101; PP944938-PP944992; PP947812-PP947814).\u003c/p\u003e \u003cp\u003eFlea infestation prevalence of cats was defined as the proportion of sampled cats that harbored at least one flea. Flea burden was defined as the number of fleas per infested host. To test for associations between cat demographic factors (sex, geographic origin) and season with the outcomes of flea burden and/or flea infection, we used Chi-square, Fisher\u0026rsquo;s exact, Wilcoxon Rank Sum, and Kruskal-Wallis tests using the program R.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eCats from Hidalgo and Cameron counties in Texas were sampled from January-June 2017. Cats were from 14 different urban locations, with most coming from the cities of McAllen (66), Pharr (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e), Edinburg (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e), and San Juan (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e; Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Of the 167 cats in the study, 87 were female and 80 were male. Most cats were sampled in the winter months (72%), fewer in spring (17%) and summer (11%) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eInfection of fleas removed from stray cats and cat blood with \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e across 14 locations of the Rio Grande Valley of south Texas, 2017. Cat blood tested for \u003cem\u003eRickettsia\u003c/em\u003e had one positive (F112).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. of Cats\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFlea infested cats (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo. flea pools tested\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eCandidatus\u003c/em\u003e Rickettsia senegalensis positive flea pools (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eRickettsia typhi\u003c/em\u003e positive flea pools (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eBartonella henselae\u003c/em\u003e positive flea pools (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eBartonella clarridgeiae\u003c/em\u003e positive flea pools (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003eBartonella henselae\u003c/em\u003e positive cat blood (% cats)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003eBartonella clarridgeiae\u003c/em\u003e positive cat blood (% cats)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLocation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlamo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8(80)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6(75)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlton\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2(67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2(67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDonna\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEdinburg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14(70)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eElsa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHarlingen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMcAllen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55(83)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10(\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4(\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e16(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5(\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMercedes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMission\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(60)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMonte Alto\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePalmview\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2(\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePharr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22(79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2(\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14(64)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10(\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSan Juan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16(89)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4(\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e9(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWeslaco\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2(100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeason\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpring\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25(89)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8(\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10(\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSummer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13(68)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6(\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWinter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e101(84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25(\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e45(\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e26(\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67(77)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8(\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e32(\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e19(\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e73(91)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20(\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e27(\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18(\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e139(83)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e140\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e28(\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e59(\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3(\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e37(\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eA total of 721 fleas were collected from 139 cats resulting in an infestation prevalence of 83% (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Flea burden ranged from 1 to 41. An average of 4.3 fleas per cat (5.2 fleas per infested cat) were found. Significantly more male than female cats harbored fleas, with 91% and 76% of male and female cats infested, respectively (Wilcoxon Rank Sum Test, W\u0026thinsp;=\u0026thinsp;2607, p-value\u0026thinsp;=\u0026thinsp;0.004). Infestation prevalence was lowest in the summer months, with fleas apparent on 101 of 120 (84%) of cats sampled in winter, 25 of 28 (89%) in spring and 13 of 19 (68%) in summer (Kruskal-Wallis test, H\u0026thinsp;=\u0026thinsp;12.43, dƒ = 2, p-value\u0026thinsp;=\u0026thinsp;0.002). Most fleas (n\u0026thinsp;=\u0026thinsp;718) were \u003cem\u003eC. felis\u003c/em\u003e (cat flea), with two \u003cem\u003eEchidnophaga gallinacea\u003c/em\u003e (sticktight flea) and one single \u003cem\u003ePulex sp\u003c/em\u003e. Most infested cats harbored only a single flea species, but cat F63 harbored a single \u003cem\u003eE. gallinacea\u003c/em\u003e and four \u003cem\u003eC. felis\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eOf 140 flea pools tested for \u003cem\u003eRickettsia\u003c/em\u003e, 34 (24%) were positive of which 28 (20% of all flea pools) were positive for \u003cem\u003eCandidatus\u003c/em\u003e Rickettsia senegalensis and 6 (4% of all flea pools) were positive for \u003cem\u003eR. typhi\u003c/em\u003e. \u003cem\u003eRickettsia\u003c/em\u003e-positive fleas were collected from 9 of the 14 locations, including McAllen (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e), four each from Alamo, Edinburg and San Juan, three from Pharr, two from Alton and one each from Donna, Mission and Palmview. \u003cem\u003eRickettsia\u003c/em\u003e infection did not show association with location (Chi-square test, χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;10.132, dƒ = 13, p\u0026thinsp;=\u0026thinsp;0.683). \u003cem\u003eRickettsia\u003c/em\u003e infection was positively associated with flea burden (Wilcoxon Rank Sum Test, W\u0026thinsp;=\u0026thinsp;968, p-value\u0026thinsp;=\u0026thinsp;1.968e-07). Further, \u003cem\u003eRickettsia\u003c/em\u003e infection prevalence in fleas was greatest in winter (30 of 34), whereas only three and one infected flea pools were found in summer and spring, respectively (Chi-square test, χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;6.607, dƒ = 2, p\u0026thinsp;=\u0026thinsp;0.0368). A total of 11 of 67 flea pools (16%) from female cats and 23 of 73 flea pools (32%) from male cats were positive for \u003cem\u003eRickettsia\u003c/em\u003e (Chi-square test, χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;5.885, dƒ = 1, p\u0026thinsp;=\u0026thinsp;0.0153). Of the 6 \u003cem\u003eR. typhi\u003c/em\u003e-positive flea pools, four were from McAllen and one each were from Pharr and Alamo (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAdditionally, non-target amplification of \u003cem\u003eBartonella\u003c/em\u003e occurred with two \u003cem\u003eRickettsia\u003c/em\u003e assays, as one flea pool (F72) amplified using the RrCS \u003cem\u003egltA\u003c/em\u003e primers showed a \u003cem\u003eB. clarridgeiae\u003c/em\u003e sequence; three flea pools amplified using the RpCS \u003cem\u003egltA\u003c/em\u003e primers showed \u003cem\u003eB. henselae\u003c/em\u003e sequences; and five flea pools amplified using the RpCS \u003cem\u003egltA\u003c/em\u003e primers resulted in genus-level identifications to \u003cem\u003eBartonella\u003c/em\u003e. All the pools that showed \u003cem\u003eBartonella\u003c/em\u003e from these two assays were also detected by the \u003cem\u003eBartonella\u003c/em\u003e-specific \u003cem\u003epap31\u003c/em\u003e PCR (Additional file 1: Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePCR amplification of the \u003cem\u003eBartonella pap 31\u003c/em\u003e locus detected was positive in 90 flea pools (64% of all pools), of which 59 (42% of all pools) were confirmed \u003cem\u003eB. henselae\u003c/em\u003e, 3 (2% of all pools) were \u003cem\u003eB. clarridgeiae\u003c/em\u003e and another 28 were identified to the genus level as \u003cem\u003eBartonella\u003c/em\u003e. \u003cem\u003eBartonella\u003c/em\u003e status did not correlate with the location of the host (Chi-square test, χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;11.135, dƒ = 13, p\u0026thinsp;=\u0026thinsp;0.6). There were no statistical differences in seasonal distribution of \u003cem\u003eBartonella\u003c/em\u003e positivity, with 55% winter, 61% spring and 36% summer pools of each season testing positive (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;2.758;, dƒ = 2, p\u0026thinsp;=\u0026thinsp;0.252). A total of 48 pools from female cats and 42 from male cats carried \u003cem\u003eBartonella\u003c/em\u003e (Chi-square test, χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.012238, dƒ = 1, p-value\u0026thinsp;=\u0026thinsp;0.912) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLastly, coinfections of flea pools with both \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e occurred more commonly than expected by chance (Chi-square test, χ2\u0026thinsp;=\u0026thinsp;5.858, dƒ = 1, p value\u0026thinsp;=\u0026thinsp;0.0155), with 25 flea pools positive for both \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e. This included five of the six \u003cem\u003eR. typhi\u003c/em\u003e-positive flea pools which all caried \u003cem\u003eBartonella\u003c/em\u003e of which 4 were \u003cem\u003eB. henselae\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eBlood clot and heart tissue DNA from 24 cats was evaluated using the Rickettsial \u003cem\u003eompB\u003c/em\u003e locus; these included all six cats that harbored \u003cem\u003eR\u003c/em\u003e. \u003cem\u003etyphi\u003c/em\u003e-positive fleas, 11 cats that harbored fleas with \u003cem\u003eCandidatus\u003c/em\u003e Rickettsia senegalensis and seven cats with \u003cem\u003eRickettsia\u003c/em\u003e-negative fleas. An expanded set of other tissues was evaluated from one cat with \u003cem\u003eR\u003c/em\u003e. \u003cem\u003etyphi\u003c/em\u003e-positive fleas (F159) and three with negative fleas (F154, F156 and F157). Although PCR products using \u003cem\u003eompB\u003c/em\u003e, \u003cem\u003eRrCS\u003c/em\u003e and in one case \u003cem\u003ehtrA\u003c/em\u003e were observed in some of these samples, only one instance (F112 blood clot) yielded sequence matching \u003cem\u003eCandidatus\u003c/em\u003e R. senegalensis. Others did not match \u003cem\u003eRickettsia\u003c/em\u003e or yielded poor quality sequences(Additional file 1: Table S2), and thus the cats were considered negative.\u003c/p\u003e \u003cp\u003eWe examined DNA extracted from blood clots of all 167 cats for \u003cem\u003eBartonella\u003c/em\u003e using PCR. Thirty-nine samples on the \u003cem\u003epap31\u003c/em\u003e PCR were positive \u0026ndash; 37 (22% of cats) carried \u003cem\u003eBartonella henselae\u003c/em\u003e, one (0.6%) carried \u003cem\u003eBartonella clarridgeiae\u003c/em\u003e and one was only identifiable to genus. When \u003cem\u003eBartonella\u003c/em\u003e in cats and fleas were compared, 30 flea pools and matching cats showed carriage of \u003cem\u003eBartonella\u003c/em\u003e (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;8.595, dƒ = 1, p\u0026thinsp;=\u0026thinsp;0.0034). The proportion of cats carrying \u003cem\u003eBartonella\u003c/em\u003e was highest in cats from San Juan (56%). Seasonally, 28 of 120 (23%) cats sampled in winter showed \u003cem\u003eBartonella\u003c/em\u003e (26 \u003cem\u003eB. henselae\u003c/em\u003e, one \u003cem\u003eB. clarridgeiae\u003c/em\u003e, one \u003cem\u003eBartonella\u003c/em\u003e spp.) while 10 of 28 (36%) in spring (all \u003cem\u003eB. henselae\u003c/em\u003e) and one of 19 (5%) in summer (one \u003cem\u003eB. henselae\u003c/em\u003e). The \u003cem\u003eB. henselae\u003c/em\u003e infection prevalence was 24% in both female (21/87) and male (19/80) cats.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study strengthens evidence that south Texas harbors robust enzootic, cat flea-driven cycles of transmission of \u003cem\u003eR. typhi\u003c/em\u003e, associated with the emergence of murine typhus in humans in the area. Further, we provide new evidence of enzootic cycles of \u003cem\u003eB. henselae\u003c/em\u003e transmission.\u003c/p\u003e \u003cp\u003eIn a recent study from Galveston, TX (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e), \u003cem\u003eR. typhi\u003c/em\u003e was detected in fleas from one of 24 feral cats (4.1%), remarkably similar to 4.3% of 140 flea pools we report here. The Galveston study also detected \u003cem\u003eCandidatus\u003c/em\u003e R. senegalensis as we did, in addition to two species that were not apparent in our study, \u003cem\u003eR. asemboensis\u003c/em\u003e and \u003cem\u003eR. felis\u003c/em\u003e. In contrast, \u003cem\u003eR. typhi\u003c/em\u003e was not detected in a survey of fleas from 283 feral/stray cats from southeastern Georgia, USA, in which 16.5% of flea pools were positive for the 17-kDa protein antigen gene of \u003cem\u003eRickettsia\u003c/em\u003e spp., none of which were positive for \u003cem\u003eompB\u003c/em\u003e of \u003cem\u003eR. typhi\u003c/em\u003e (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDespite the relatively high infection prevalence of \u003cem\u003eBartonella\u003c/em\u003e we detected in both vectors (64% of flea pools) and hosts (23% of cats), relatively little ecological work has been done on this agent in the southern USA. In southeastern Georgia, USA, 35.2% of flea pools from feral/stray cats were infected with \u003cem\u003eBartonella\u003c/em\u003e (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e). In a study of cats from catteries across North America, a seroprevalence of 35.8% was recorded, in which flea infestation was the most important risk factor for high \u003cem\u003eB. henselae\u003c/em\u003e seroprevalence (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Lower \u003cem\u003eBartonella\u003c/em\u003e infection prevalence among fleas removed from cats have been reported in studies from Canada (4.3%), Chile (7.5%), Ethiopia (6%) and Greece (13.5%) (\u003cspan additionalcitationids=\"CR41 CR42\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAs expected, we found cats with \u003cem\u003eBartonella\u003c/em\u003e-positive blood commonly harbored infected fleas, yet there were also several infected flea pools removed from cats whose blood and tissues tested negative, suggesting that those fleas may have picked up the infection from a prior bloodfeeding event on a different host. Similarly, a study of cats in Brazil found \u003cem\u003eBartonella\u003c/em\u003e DNA in 47.8% of the cat blood samples and 18.3% of \u003cem\u003eC. felis\u003c/em\u003e fleas, in which cats infested by positive ectoparasites showed approximately twice the odds of being infected (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTwo cats (F31 and F33) simultaneously harbored fleas infested with both \u003cem\u003eR. typhi\u003c/em\u003e and \u003cem\u003eB. henselae\u003c/em\u003e, potentially serving as companion animal sentinels of multiple regional zoonotic threats to humans. Further, this exact sample set of cats has also been examined in previous studies for additional zoonoses, including \u003cem\u003eTrypanosoma cruzi\u003c/em\u003e, the causal agent of Chagas disease (19/167 cats, 11.4% prevalence) and \u003cem\u003eDirofilaria immitis\u003c/em\u003e (canine heartworm) which occasionally causes pulmonary disease in humans (22/122 cats, 18% prevalence) (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e). In comparing the results across studies, two cats (F96 and F141) were infected with \u003cem\u003eBartonella\u003c/em\u003e, \u003cem\u003eT. cruzi\u003c/em\u003e and \u003cem\u003eD. immitis\u003c/em\u003e; five (F96, F101, F133, F141 and F142) with \u003cem\u003eBartonella\u003c/em\u003e and \u003cem\u003eT. cruzi\u003c/em\u003e; and eight with \u003cem\u003eBartonella\u003c/em\u003e and \u003cem\u003eD. immitis\u003c/em\u003e. One \u003cem\u003eT. cruzi\u003c/em\u003e-positive cats harbored \u003cem\u003eR. typhi\u003c/em\u003e-positive fleas (F35), and two \u003cem\u003eT. cruzi\u003c/em\u003e-positive cats harbored \u003cem\u003eCandidatus\u003c/em\u003e R. senegalensis-positive fleas (F26 and F113). While an older study suggested pet ownership is not a direct risk factor for human exposure to \u003cem\u003eR. typhi\u003c/em\u003e based on Texas data (\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e) owning a dog was found to be a risk factor for murine typhus seropositivity in a cross-sectional study from India (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e). While feral/stray cats that remain outdoors may pose little risk of zoonoses transmission from flea importation to the house, those cats that enter homes and those that interact with domestic animals may facilitate \u0026lsquo;zoonoses in the bedroom\u0026rsquo; by introducing new disease threats to the domestic setting (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCases of murine typhus have been steadily increasing in the United States over the past three decades with 348 cases from 1990-99, 994 from 2000-09, a stark 3159 from 2010\u0026ndash;2018. A total of 6700 cases were reported in Texas between 2008-23 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.dshs.texas.gov\u003c/span\u003e\u003cspan address=\"https://www.dshs.texas.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Within Texas, the Rio Grande Valley appears to be a hotspot for local cases where Hidalgo County- the location of this current study- reported 1383 cases between 2008 to 2023 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.dshs.texas.gov\u003c/span\u003e\u003cspan address=\"https://www.dshs.texas.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). In an 18-year period from 1998 to 2016, 213 pediatric patients were admitted to a hospital in south Texas with a confirmed diagnosis of murine typhus (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNationally, cat scratch disease is diagnosed for 4.5\u0026ndash;9.3 patients per 100,000 people, highest in the southern United States, in children aged 5\u0026ndash;9, and with hospitalization higher among males 50\u0026ndash;64 years old (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e). In a survey across 9 years of nearly 40\u0026nbsp;million medical insurance enrollees, 0.03% were diagnosed with cat scratch disease with nearly one-fifth (19.7%) of cases from the West South Central US census region that includes Texas, second to 26.3% in the South Atlantic US census region (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). In a similar analysis, over a ten-year period, over half the diagnosis of cat scratch disease was from a southern state in the US (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). Although it is established that \u003cem\u003eBartonella\u003c/em\u003e infections are life threatening for immunocompromised individuals, neurobartonellosis in immunocompetent individuals is incompletely understood (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e) and the public health burden of bartonellosis is unknown. Cats are deemed to be adapted to \u003cem\u003eB. henselae\u003c/em\u003e but non-adapted species of \u003cem\u003eBartonella\u003c/em\u003e and occasional variants of \u003cem\u003eB. henselae\u003c/em\u003e can cause cardiac disease in cats (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNew \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e species and novel vector-host associations continue to be described (\u003cspan additionalcitationids=\"CR51\" citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e), underscoring the need for vector and host surveillance to provide an ecological basis for public and veterinary health risk assessment. Controlling fleas and infections in domestic cats may help to alleviate spillover transmission to humans.\u003c/p\u003e"},{"header":"Declarations","content":" \u003cp\u003e \u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e \u003cp\u003eThe Texas A\u0026amp;M. University Institutional Animal Care and Use Committee (IACUC) issued a letter of exemption on December 16, 2016, as samples were collected exclusively from cats that were euthanized for non-study reasons.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eConsent for publication\u003c/strong\u003e \u003cp\u003eNot Applicable\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eCompeting interests\u003c/h2\u003e \u003cp\u003eThe authors declare that they have no competing interests\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eTexas A\u0026amp;M AgriLife Research provided funding.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eSB: conceptualization; data curation; formal analysis; investigation; writing- original draft; writing- review \u0026amp;amp; editing.IZ: conceptualization; investigation; writing- review \u0026amp;amp; editing.AK: investigation; methodology; writing- review \u0026amp;amp; editing.SH: conceptualization; funding acquisition; investigation; project administration; resources; writing- review \u0026amp;amp; editing.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe thank Lisa Auckland and Keswick Killets for assistance in the laboratory and Dr. Jose Juarez-Valdez for the map for Figure 1. Graphical abstract was created in BioRender. Balasubramanian, S. (2026) https://BioRender.com/kssvz5w\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated and/or analysed during the current study are available in the OAKTrust Digital Repository [https://oaktrust.library.tamu.edu/home](https:/oaktrust.library.tamu.edu/home) .\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eFang R, Blanton LS, Walker DH. Rickettsiae as emerging infectious agents. Clin Lab Med. 2017;37(2):383\u0026ndash;400.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKyle G. Rodino. Rickettsioses in the United States | Elsevier Enhanced Reader. Clin Microbiol Newsl. 2019;41(13):113\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAzad AF, Radulovic S, Higgins JA, Noden BH, Troyer JM. Flea-borne rickettsioses: ecologic considerations. Emerg Infect Dis. 1997;3(3):319\u0026ndash;27.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eP\u0026eacute;rez-Osorio CE, Zavala-Vel\u0026aacute;zquez JE, Le\u0026oacute;n JJA, Zavala-Castro JE. \u003cem\u003eRickettsia felis\u003c/em\u003e as Emergent Global Threat for Humans. Emerg Infect Dis. 2008;14(7):1019\u0026ndash;23.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnstead GM. History, rats, fleas, and opossums. II. The decline and resurgence of flea-borne typhus in the United States, 1945\u0026ndash;2019. Trop Med Infect Dis. 2020;6(1):2.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCiven R, Ngo V. Murine typhus: An unrecognized suburban vectorborne disease. Clin Infect Dis. 2008;46(6):913\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCaravedo Martinez MA, Ram\u0026iacute;rez-Hern\u0026aacute;ndez A, Blanton LS. Manifestations and management of flea-borne Rickettsioses. Res Rep Trop Med. 2021;Volume 12:1\u0026ndash;14.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJuhasz NB, Wilson JM, Haney KN, Clark MH, Davenport AC, Breitschwerdt EB, et al. \u003cem\u003eRickettsia typhi\u003c/em\u003e infection in a clinically-ill dog from Houston, Texas. Vet Parasitol Reg Stud Rep. 2022;35:100781.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSorvillo FJ, Gondo B, Emmons R, Ryan P, Waterman SH, Tilzer A, et al. A suburban focus of endemic typhus in Los Angeles county: association with seropositive domestic cats and opossums. Am J Trop Med Hyg. 1993;48(2):269\u0026ndash;73.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGracia MJ, Marc\u0026eacute;n JM, Pinal R, Calvete C, Rodes D. Prevalence of \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e species in Spanish cats and their fleas. J Vector Ecol. 2015;40(2):233\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChomel BB, Boulouis HJ, Breitschwerdt EB, Kasten RW, Vayssier-Taussat M, Birtles RJ, et al. Ecological fitness and strategies of adaptation of \u003cem\u003eBartonella\u003c/em\u003e species to their hosts and vectors. Vet Res. 2009;40(2):29.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNelson CA, Saha S, Mead PS. Cat-Scratch disease in the United States, 2005\u0026ndash;2013. Emerg Infect Dis. 2016;22(10):1741\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKordick DL, Papich MG, Breitschwerdt EB. Efficacy of enrofloxacin or doxycycline for treatment of \u003cem\u003eBartonella henselae\u003c/em\u003e or \u003cem\u003eBartonella clarridgeiae\u003c/em\u003e infection in cats. Antimicrob Agents Chemother. 1997;41(11):2448\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJacomo V, Kelly PJ, Raoult D. Natural History of \u003cem\u003eBartonella\u003c/em\u003e Infections (an Exception to Koch\u0026rsquo;s Postulate). Clin Vaccine Immunol. 2002;9(1):8\u0026ndash;18.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJurja S, Stroe AZ, Pundiche MB, Docu Axelerad S, Mateescu G, Micu AO, et al. The clinical profile of cat-scratch disease\u0026rsquo;s neuro-ophthalmological effects. Brain Sci. 2022;12(2):217.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNawrocki CC, Max RJ, Marzec NS, Nelson CA. Atypical manifestations of cat-scratch disease, United States, 2005\u0026ndash;2014 - Volume 26, Number 7\u0026mdash;July 2020 - Emerging Infectious Diseases journal - CDC. Emerg Infect Dis [Internet]. 2020 Jul [cited 2022 Sep 20];26(7). Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://wwwnc.cdc.gov/eid/article/26/7/20-0034_article\u003c/span\u003e\u003cspan address=\"https://wwwnc.cdc.gov/eid/article/26/7/20-0034_article\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDelaney S, Robveille C, Maggi RG, Lashnits E, Kingston E, Liedig C, et al. Bartonella species bacteremia in association with adult psychosis. Front Psychiatry. 2024;15:1388442.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRolain JM, La Scola B, Liang Z, Davoust B, Raoult D. Immunofluorescent detection of intraerythrocytic \u003cem\u003eBartonella henselae\u003c/em\u003e in naturally infected cats. J Clin Microbiol. 2001;39(8):2978\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKordick DL, Brown TT, Shin K, Breitschwerdt EB. Clinical and pathologic evaluation of chronic \u003cem\u003eBartonella henselae\u003c/em\u003e or \u003cem\u003eBartonella clarridgeiae\u003c/em\u003e infection in cats. J Clin Microbiol. 1999;37(5):1536\u0026ndash;47.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBreitschwerdt EB, Lappin MR. Feline bartonellosis: we\u0026rsquo;re just scratching the surface. J Feline Med Surg. 2012;14(9):609\u0026ndash;10.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlanton LS, Vohra RF, Fistein L, Quade B, Walker DH, Bouyer DH. Rickettsiae within the fleas of feral cats in Galveston, Texas. Vector-Borne Zoonotic Dis. 2019;19(9):647\u0026ndash;51.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuang HHH, Power RI, Mathews KO, Ma GC, Bosward KL, Šlapeta J. Cat fleas (Ctenocephalides felis clade \u0026lsquo;Sydney\u0026rsquo;) are dominant fleas on dogs and cats in New South Wales, Australia: Presence of flea-borne \u003cem\u003eRickettsia felis\u003c/em\u003e, \u003cem\u003eBartonella\u003c/em\u003e spp. but absence of \u003cem\u003eCoxiella burnetii\u003c/em\u003e DNA. Curr Res Parasitol Vector-Borne Dis. 2021;1:100045.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiodaki M, Spanakos G, Samarkos M, Daikos GL, Christopoulou V, Piperaki ET. Molecular screening of cat and dog ectoparasites for the presence of Bartonella spp. in Attica, Greece. Acta Vet Hung. 2022;\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaimundo JM, Guimar\u0026atilde;es A, Amaro GM, Silva AT da, Rodrigues CJBC, Santos HA, et al. Prevalence of Bartonella species in shelter cats and their ectoparasites in southeastern Brazil. Rev Bras Parasitol Veterin\u0026aacute;ria. 2022;31(1):e014221.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRazgūnaitė M, Lipatova I, Paulauskas A, Karvelienė B, Riškevičienė V, Radzijevskaja J. \u003cem\u003eBartonella\u003c/em\u003e infections in cats and cat fleas in Lithuania. Pathogens. 2021;10(9):1209.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBitam I, Dittmar K, Parola P, Whiting MF, Raoult D. Fleas and flea-borne diseases. Int J Infect Dis. 2010;14(8):e667\u0026ndash;76.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFoley JE, Chomel B, Kikuchi Y, Yamamoto K, Pedersen NC. Seroprevalence of \u003cem\u003eBartonella henselae\u003c/em\u003e in cattery cats: Association with cattery hygiene and flea infestation. Vet Q. 1998;20(1):1\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIannino. Fleas of dog and cat: species, biology and flea-borne diseases. Vet Ital. 2017;53(4):273\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZecca IB, Hodo CL, Slack S, Auckland L, Rodgers S, Killets KC, et al. Prevalence of \u003cem\u003eTrypanosoma cruzi\u003c/em\u003e infection and associated histologic findings in domestic cats (Felis catus). Vet Parasitol. 2020;278:109014.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLawrence AL, Webb CE, Clark NJ, Halajian A, Mihalca AD, Miret J, et al. Out-of-Africa, human-mediated dispersal of the common cat flea, \u003cem\u003eCtenocephalides felis\u003c/em\u003e: The hitchhiker\u0026rsquo;s guide to world domination. Int J Parasitol. 2019;49(5):321\u0026ndash;36.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCenters for Disease Control. Pictorial keys to arthropods, reptiles, birds, and mammals of public health significance [Internet]. 1966 [cited 2025 Dec 15]. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://stacks.cdc.gov\u003c/span\u003e\u003cspan address=\"https://stacks.cdc.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFolmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol. 1994;3(5):294\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoux V, Raoult D. Phylogenetic analysis of members of the genus \u003cem\u003eRickettsia\u003c/em\u003e using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol. 2000;50(4):1449\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWilliamson PC, Billingsley PM, Teltow GJ, Seals JP, Turnbough MA, Atkinson SF. \u003cem\u003eBorrelia\u003c/em\u003e, \u003cem\u003eEhrlichia\u003c/em\u003e, and \u003cem\u003eRickettsia\u003c/em\u003e spp. in ticks removed from persons, Texas, USA. Emerg Infect Dis. 2010;16(3):441\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRegenery RL, Spruill CL, Plikaytis BD. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes [Internet]. 1991 [cited 2022 Mar 1]. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://journals.asm.org/doi/epdf/\u003c/span\u003e\u003cspan address=\"https://journals.asm.org/doi/epdf/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/jb.173.5.1576-1589.1991\u003c/span\u003e\u003cspan address=\"10.1128/jb.173.5.1576-1589.1991\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWebb L, Mitchell C, Malloy DC, Dasch GA, Azad AF. Detection of murine typhus infection in fleas by using the polymerase chain reaction [Internet]. 1990 [cited 2022 Mar 1]. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://journals.asm.org/doi/epdf/\u003c/span\u003e\u003cspan address=\"https://journals.asm.org/doi/epdf/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/jcm.28.3.530-534.1990\u003c/span\u003e\u003cspan address=\"10.1128/jcm.28.3.530-534.1990\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeaiter Z, Fournier PE, Ogata H, Raoult D. Phylogenetic classification of \u003cem\u003eBartonella\u003c/em\u003e species by comparing groEL sequences. Int J Syst Evol Microbiol. 2002;52(1):165\u0026ndash;71.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33(7):1870\u0026ndash;4.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown LD, Maness R, Greer K. Detection of \u003cem\u003eBartonella\u003c/em\u003e spp. and \u003cem\u003eRickettsia\u003c/em\u003e spp. in cat fleas (\u003cem\u003eCtenocephalides felis\u003c/em\u003e) collected from free-roaming domestic cats in southeastern Georgia, USA. Vet Parasitol Reg Stud Rep. 2022;100743.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiodaki M, Spanakos G, Samarkos M, Daikos GL, Christopoulou V, Piperaki ET. Molecular screening of cat and dog ectoparasites for the presence of \u003cem\u003eBartonella\u003c/em\u003e spp. in Attica, Greece. Acta Vet Hung. 2022;70(1):9\u0026ndash;14.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM\u0026uuml;ller A, Rodr\u0026iacute;guez E, Walker R, Bittencourt P, P\u0026eacute;rez-Macchi S, Gon\u0026ccedil;alves LR, et al. Occurrence and genetic diversity of \u003cem\u003eBartonella\u003c/em\u003e spp. (Rhizobiales: Bartonellaceae) and \u003cem\u003eRickettsia\u003c/em\u003e spp. (Rickettsiales: Rickettsiaceae) in cat fleas (Siphonaptera: Pulicidae) From Chile. J Med Entomol. 2018;55(6):1627\u0026ndash;32.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumsa B, Parola P, Raoult D, Socolovschi C. Molecular Detection of \u003cem\u003eRickettsia felis\u003c/em\u003e and \u003cem\u003eBartonella henselae\u003c/em\u003e in dog and cat fleas in central Oromia, Ethiopia. Am J Trop Med Hyg. 2014;90(3):457\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKamrani A, Parreira VR, Greenwood J, Prescott JF. The prevalence of Bartonella, hemoplasma, and \u003cem\u003eRickettsia felis\u003c/em\u003e infections in domestic cats and in cat fleas in Ontario. Can J Vet Res. 2008;72(5):411\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMosley IA, Zecca IB, Tyagi N, Harvey TV, Hamer SA, Verocai GG. Occurrence of \u003cem\u003eDirofilaria immitis\u003c/em\u003e infection in shelter cats in the lower Rio Grande Valley region in South Texas, United States, using integrated diagnostic approaches. Vet Parasitol Reg Stud Rep. 2023;41:100871.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWiggers RJ, Stewart RS. Ownership of cats or dogs does not increase exposure to \u003cem\u003eRickettsia typhi\u003c/em\u003e. Tex Med. 2002;98(6):56\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDevamani CS, Schmidt WP, Ariyoshi K, Anitha A, Kalaimani S, Prakash JAJ. Risk factors for scrub typhus, murine typhus, and spotted fever seropositivity in urban areas, rural plains, and peri-forest hill villages in south India: a cross-sectional study. Am J Trop Med Hyg. 2020;103(1):238\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChomel BB, Sun B. Zoonoses in the bedroom. Emerg Infect Dis. 2011;17(2):167\u0026ndash;72.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHoward A, Fergie J. Murine Typhus in South Texas Children: An 18-year Review. Pediatr Infect Dis J. 2018;37(11):1071\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNelson CA, Moore AR, Perea AE, Mead PS. Cat scratch disease: U.S. clinicians\u0026rsquo; experience and knowledge. Zoonoses Public Health. 2018;65(1):67\u0026ndash;73.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003edo Amaral RB, Cardozo MV, Varani A de M, Furquim MEC, Dias CM, Assis WO de, et al. First report of \u003cem\u003eBartonella\u003c/em\u003e spp. in marsupials from Brazil, with a description of \u003cem\u003eBartonella harrusi\u003c/em\u003e sp. nov. and a new proposal for the taxonomic reclassification of species of the genus \u003cem\u003eBartonella\u003c/em\u003e. Microorganisms. 2022;10(8):1609.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMedkour H, Lo CI, Anani H, Fenollar F, Mediannikov O. \u003cem\u003eBartonella massiliensis\u003c/em\u003e sp. nov., a new bacterial species isolated from an \u003cem\u003eOrnithodoros sonrai\u003c/em\u003e tick from Senegal. New Microbes New Infect. 2019;32:100596.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLaroche M, Berenger JM, Mediannikov O, Raoult D, Parola P. Detection of a Potential New \u003cem\u003eBartonella\u003c/em\u003e Species \u0026ldquo;\u003cem\u003eCandidatus\u003c/em\u003e Bartonella rondoniensis\u0026rdquo; in Human Biting Kissing Bugs (Reduviidae; Triatominae). G\u0026uuml;rtler RE, editor. PLoS Negl Trop Dis. 2017;11(1):e0005297.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"parasites-and-vectors","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"parv","sideBox":"Learn more about [Parasites \u0026 Vectors](http://parasitesandvectors.biomedcentral.com/)","snPcode":"13071","submissionUrl":"https://submission.nature.com/new-submission/13071/3","title":"Parasites \u0026 Vectors","twitterHandle":"@bugbittentweets","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Ctenocephalides felis, Rickettsia typhi, murine typhus, Bartonella henselae, cat scratch disease, Rio Grande Valley, Texas","lastPublishedDoi":"10.21203/rs.3.rs-8726915/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8726915/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFlea-borne rickettsiosis and bartonellosis are emerging diseases in humans and animals. There is increasing concern for the emergence of \u003cem\u003eRickettsia typhi\u003c/em\u003e (agent of murine typhus or flea-borne typhus) in humans of southern USA, for which cats are key reservoirs and cat fleas (\u003cem\u003eCtenocephalides felis\u003c/em\u003e) serve as the vector. Similarly, multiple species of \u003cem\u003eBartonella\u003c/em\u003e have cats as natural hosts and fleas as vectors, leading to varied disease outcomes in humans, including cat scratch disease and neurobartonellosis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate the ecology of \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e pathogens, we studied 167 predominantly stray cats (\u003cem\u003eFelis catus\u003c/em\u003e) from the Rio Grande Valley in south Texas. Fleas were collected, identified morphologically and confirmed molecularly. DNA from fleas was tested for \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e using PCR and Sanger sequencing of multiple gene targets. Additionally, DNA from blood of all cats and tissues from a subset of cats were assayed for infections.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFlea infestation prevalence of cats was 83% - higher in male vs. female cats - yielding 721 fleas, predominantly \u003cem\u003eC. felis\u003c/em\u003e with 2 \u003cem\u003eEchidnophaga gallinacea\u003c/em\u003e and 1 \u003cem\u003ePulex sp\u003c/em\u003e. Flea burden was a significant predictor of Rickettsia infection in fleas. \u003cem\u003eCandidatus\u003c/em\u003e Rickettsia senegalensis was identified in 28 flea pools (20% of all pools) and \u003cem\u003eRickettsia typhi\u003c/em\u003e in 6 flea pools (4%). \u003cem\u003eBartonella henselae\u003c/em\u003e was identified in 59 flea pools (42% of all pools) and blood from 37 cats (22.2%); \u003cem\u003eBartonella clarridgeiae\u003c/em\u003e was detected in 3 flea pools (2% of pools) and blood from 1 cat (0.7%). Thirty cats had simultaneously \u003cem\u003eBartonella-\u003c/em\u003epositive fleas and blood. \u003cem\u003eRickettsia\u003c/em\u003e infection was positively associated with flea burden. Co-infections of \u003cem\u003eRickettsia\u003c/em\u003e and \u003cem\u003eBartonella\u003c/em\u003e in fleas occurred more commonly than expected by chance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCats and their fleas support robust transmission of \u003cem\u003eR. typhi\u003c/em\u003e, \u003cem\u003eB. henselae \u003c/em\u003eand related agents in south Texas with implications for veterinary and public health.\u003c/p\u003e","manuscriptTitle":"Rickettsia typhi, Bartonella henselae and related zoonotic agents in fleas from domestic cats (Felis catus) from the Rio Grande Valley, Texas","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-04 08:30:17","doi":"10.21203/rs.3.rs-8726915/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-04T23:47:36+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-03T12:43:50+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-03T05:07:39+00:00","index":"","fulltext":""},{"type":"submitted","content":"Parasites \u0026 Vectors","date":"2026-01-29T04:13:17+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"parasites-and-vectors","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"parv","sideBox":"Learn more about [Parasites \u0026 Vectors](http://parasitesandvectors.biomedcentral.com/)","snPcode":"13071","submissionUrl":"https://submission.nature.com/new-submission/13071/3","title":"Parasites \u0026 Vectors","twitterHandle":"@bugbittentweets","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"599597c0-cc87-4529-ab8e-b157f4b4700f","owner":[],"postedDate":"February 4th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-13T14:25:56+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-04 08:30:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8726915","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8726915","identity":"rs-8726915","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}