Background Over the last 15 years, visceral leishmaniasis (VL) has emerged as a public health concern in Tbilisi, the capital of Georgia. Methodology/Principal Findings Seroepidemiological surveys were conducted to determine the prevalence and incidence of infection in children and dogs within the main focus of VL, and to identify risk factors associated with human infection. Of 4,250 children investigated, 7.3% were positive by direct agglutination test in a baseline survey; an apparent incidence rate of 6.0% was estimated by one year follow-up. None of the seropositive children progressed to VL during the survey. Increased seropositivity at one year was predicted by presence at baseline of clustered flying insects (OR = 1.49; P = 0.001), perceived satisfactory sanitation (OR = 1.65; P<0.001), stray dogs (OR = 1.33; P = 0.023), and by persistent fever during the 6 months prior to baseline survey (OR = 14.2; P<0.001). Overall, 18.2% (107/588) of domestic and 15.3% (110/718) of stray dogs were seropositive by the rk39 dipstick test. Clinical VL signs were found in 1.3% of domestic and 2.9% of stray, seropositive dogs. Parasites isolated from human and dog samples were identified by PCR and phylogenetic analysis of the Leishmania 70 kDa heat-shock protein (HSP70) gene as Leishmania infantum. Conclusions/Significance There is an active focus of L. infantum transmission in Tbilisi with a high prevalence of human and canine infections.
References
[1]
World Health Organization (WHO) (1996) Manual of Visceral Leishmaniasis Control. Division of Control of Tropical Diseases. Geneva, Switzerland: WHO.
[2]
Dereure J, El-Safi SH, Bucheton B, Boni M, Kheir MM, et al. (2003) Visceral leishmaniasis in eastern Sudan: parasite identification in humans and dogs; host-parasite relationships. Microbes Infect 5: 1103–1108. doi: 10.1016/j.micinf.2003.07.003
[3]
Romero GAS, Boelaert M (2010) Control of Visceral Leishmaniasis in Latin America – A Systematic Review. PLoS Negl Trop Dis 4: e584. doi: 10.1371/journal.pntd.0000584
[4]
Dujardin J-C, Campino L, Ca?avate C, Dedet J-P, Gradoni L, et al. (2008) Spread of Vector-borne Diseases and Neglect of Leishmaniasis, Europe. Emerg Infect Dis 14: 1013–1018. doi: 10.3201/eid1407.071589
[5]
Solano-Gallego L, Morell P, Arboix M, Alberola J, Ferrer L (2001) Prevalence of Leishmania infantum Infection in Dogs Living in an Area of Canine Leishmaniasis Endemicity Using PCR on Several Tissues and Serology. J Clin Microbiol 39: 560–563. doi: 10.1128/JCM.39.2.560-563.2001
[6]
Papadopoulou C, Kostoula A, Dimitriou D, Panagiou A, Bobojianni C, et al. (2005) Human and canine leishmaniasis in asymptomatic and symptomatic population in Northwestern Greece. J Infection 50: 53–60. doi: 10.1016/j.jinf.2004.05.004
[7]
Maroli M, Krasnonos L, Gafurov I (2001) Epidemiological and entomological survey in a focus of visceral leishmaniasis in Pap district (Fergana Valley) of Namangan region, Uzbekistan. Acta Trop 80: 223–228. doi: 10.1016/S0001-706X(01)00179-6
[8]
Maruashvili GM (1961) About types of visceral leishmaniasis foci. (Russian) Medical Parasitology (Moscow) 30: 188–190.
[9]
Bardjadze BG (1966) Some questions about visceral leishmaniasis in Georgia. (Georgian) Sabchota Medicina (Tbilisi) 2: 28–32.
[10]
Chubabria G, Zenaishvili O (2003) Modern concepts of visceral leishmaniasis in Georgia. (Russian) Medical Parasitology (Moscow) 2: 27–30.
[11]
Meredith SEO, Kroon NCM, Sondorp E, Seaman J, Goris MGA, et al. (1995) Leish-KIT, a Stable Direct Agglutination Test Based on Freeze-Dried Antigen for Serodiagnosis of Visceral Leishmaniasis. J Clin Microbiol 33: 1742–1745.
[12]
Abdallah KAA, Nour BYM, Schallig HDFH, Mergani A, Hamid Z, et al. (2004) Evaluation of the direct agglutination test based on freeze-dried Leishmania donovani promastigotes for the serodiagnosis of visceral leishmaniasis in Sudanese patients. Trop Med Int Health 9: 1127–1131. doi: 10.1111/j.1365-3156.2004.01308.x
[13]
Oskam L, Nieuwenhuijs JL, Hailu A (1999) Evaluation of the direct agglutination test (DAT) using freeze-dried antigen for the detection of anti-Leishmania antibodies in stored sera from various patient groups in Ethiopia. T Roy Soc Trop Med H 93: 275–277. doi: 10.1016/s0035-9203(99)90021-4
[14]
Anders G, Eisenberger C, Jonas F, Greenblatt C (2002) Distinguishing Leishmania tropica and Leishmania major in the Middle East using the polymerase chain reaction with kinetoplast DNA-specific primers. T Roy Soc Trop Med H 96: S1/87-S1192. doi: 10.1016/s0035-9203(02)90057-x
[15]
Garcia L, Kindt A, Bermudez H, Llanos-Cuentas A, De Doncker S, et al. (2004) Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. J Clin Microbiol 42: 2294–2297. doi: 10.1128/JCM.42.5.2294-2297.2004
[16]
Montalvo AM, Fraga J, Monzote L, Montano I, De Doncker S, et al. (2010) Heat-shock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World. Parasitology 137: 1159–1168. doi: 10.1017/S0031182010000089
[17]
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 4876–4882. doi: 10.1093/nar/25.24.4876
[18]
Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5: 150–163. doi: 10.1093/bib/5.2.150
[19]
Liang KY, Zeger SL (1986) Longitudinal Data Analysis Using Generalized Linear Models Biometrika 73: 13–22. doi: 10.1093/biomet/73.1.13
[20]
Yanagawa T, Gladen BC (1984) Estimating disease rates from a diagnostic test. Am J Epidemiol 119: 1015–1023.
[21]
Chappuis F, Rijal S, Soto A, Menten J, Boelaert M (2006) A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis. BMJ 333: 723. doi: 10.1136/bmj.38917.503056.7C
[22]
Agresti A (1990) Categorical Data Analysis. Wiley: New York.
[23]
Harith AE, Kolk AHJ, Leeuwenburg J, Muigai R, Huigen E (1988) Improvement of a Direct Agglutination Test for Field Studies of Visceral Leishmaniasis. J Clin Microbiol 26: 1321–1325.
[24]
Bimal S, Das VNR, Sinha PK, Gupta AK, Verma N, et al. (2005) Usefulness of the direct agglutination test in the early detection of subclinical Leishmania donovani infection: a community-based study. Ann Trop Med Parasit 99: 743–749. doi: 10.1179/136485905X65107
[25]
Schenkel K, Rijal S, Koirala S, Koirala S, Vanlerberghe V, et al. (2006) Visceral leishmaniasis in southeastern Nepal: A cross-sectional survey on Leishmania donovani infection and its risk factors. Trop Med Int Health 11: 1792–1799. doi: 10.1111/j.1365-3156.2006.01735.x
[26]
Harith AE, Slappendel RJ, Reiter I, Van Knapen F, De Korte P (1989) Application of a Direct Agglutination Test for Detection of Specific Anti-Leishmania Antibodies in the Canine Reservoir. J Clin Microbiol 27: 2252–2257.
[27]
El-Masum MA, Evans DA, Minter DM, El-Harith A (1995) Visceral leishmaniasis in Bangladesh: the value of DAT as a diagnostic tool. T Roy Soc Trop Med H 89: 185–186. doi: 10.1016/0035-9203(95)90487-5
[28]
Joshi AB, Singhasivanon P, Khusmith S, Fungladda W, Nandy A (1999) Evaluation of direct agglutination test (DAT) as an immunodiagnostic tool for diagnosis of visceral leishmaniasis in Nepal. The Southeast Asian J Trop Med Pub Health 30: 583–585.
[29]
Do?an N, B?r O, Dinleyici EC, T?z SO, Ozbel Y (2008) Investigation of anti-Leishmania seroprevalence by different serologic assays in children inhabiting in the northwestern part of Turkey. Mikrobiyol Bul 42: 103–11.
[30]
Marty P, Izri A, Ozon C, Haas P, Rosenthal E, et al. (2007) A century of leishmaniasis in Alpes-Maritimes, France. Ann Trop Med Parasit 101: 563–74. doi: 10.1179/136485907X229121
Fakhar M, Motazedian MH, Hatam GR, Asgari Q, Kalantari M, et al. (2008) Asymptomatic human carriers of Leishmania infantum: possible reservoirs for Mediterranean visceral leishmaniasis in southern Iran. Ann Trop Med Parasit 102: 577–583. doi: 10.1179/136485908X337526
[33]
Tagi-zade TA, Gasanida GB, Safianova VM, Shalmiev GB, Gadzhibekova EA (1989) Visceral leishmaniasis in Ordubad District, Nakhichevan ASSR. Medical Parasitology (Moscow) 3: 22–27.
[34]
Le Fichoux Y, Quaranta J-F, Aufeuvre J-P, Lelievre A, Marty P, et al. (1999) Occurrence of Leishmania infantum Parasitemia in Asymptomatic Blood Donors Living in an Area of Endemicity in Southern France. J Clin Microbiol 37: 1953–1957.
[35]
Costa CHN, Srewart JM, Gomes RBB (2002) Asymptomatic human carriers of Leishmania chagasi. Am J Trop Med Hyg 66: 334–337.
[36]
Martín-Sánchez J, Pineda JA, Morillas-Márquez F, García-García JA, Acedo C, et al. (2004) Detection of Leishmania infantum kinetoplast DNA in peripheral blood from asymptomatic individuals at risk for parenterally transmitted infections: relationship between PCR results and other Leishmania infection markers. Am J Trop Med Hyg 70: 545–548.
[37]
Diza E, Kansouzidou A, Gerou S, Vezyri E, Metallidis S, et al. (2008) Leishmaniases in Northern Greece: seroprevalence of the infection and incidence of the disease during the period 2001–2006. Eur J Clin Microbiol Infect Dis 27: 997–1003. doi: 10.1007/s10096-008-0538-y
[38]
Schaefer KU, Kurtzhals JA, Gachihi GS, Muller AS, Kager PA (1995) A prospective sero–epidemiological study of visceral leishmaniasis in Baringo district, Rift Valley province, Kenya. T Roy Soc Trop Med H 89: 471–475. doi: 10.1016/0035-9203(95)90070-5
[39]
Riera C, Fisa R, Udinab M, Gállegoa M, Portusa M (2004) Detection of Leishmania infantum cryptic infection in asymptomatic blood donors living in an endemic area (Eivissa, Balearic Islands, Spain) by different diagnostic methods. T Roy Soc Trop Med H 98: 102–110. doi: 10.1016/s0035-9203(03)00015-4
[40]
Badaro R, Jones TC, Carvalho EM, Sampaio D, Reed SG, et al. (1986) New perspectives on a subclinical form of visceral leishmaniasis. J Infect Dis 154: 1003–11. doi: 10.1093/infdis/154.6.1003
[41]
Gama MEA, Costa JML, Gomes CMC, Corbett CEP (2004) Subclinical Form of the American Visceral Leishmaniasis. Mem Inst Oswaldo Cruz, Rio de Janeiro 99: 889–893. doi: 10.1590/s0074-02762004000800018
[42]
Crescente JAB, Silveira FT, Lainson R, Gomes CMC, Laurenti MD, et al. (2009) A cross-sectional study on the clinical and immunological spectrum of human Leishmania (L.) infantum chagasi infection in the Brazilian Amazon region. T Roy Soc Trop Med H 103: 1250–1256. doi: 10.1016/j.trstmh.2009.06.010
[43]
Bar?o SC, Camargo-Neves VLF, Resende RM, Da Silva LJ (2007) Human Asymptomatic Infection in Visceral Leishmaniasis: A Seroprevalence Study in an Urban Area of Low Endemicity. Preliminary Results. Am J Trop Med Hyg 77: 1051–1053.
[44]
Gavgani ASM, Mohite H, Edrissian GH, Mohebali M, Davies CR (2002) Domestic dog ownership in Iran is a risk factor for human infection with Leishmania infantum. Am J Trop Med Hyg 67: 511–515.
[45]
Otranto D, Paradies P, De Caprariis D, Stanneck D, Testini G, et al. (2009) Toward Diagnosing Leishmania infantum Infection in Asymptomatic Dogs in an Area Where Leishmaniasis Is Endemic. Clinical and Vaccine Immunology 16: 337–343. doi: 10.1128/CVI.00268-08
[46]
Dantas-Torres F, Felino de Brito EM, Brand?o-Filho PS (2006) Seroepidemiological survey on canine leishmaniasis among dogs from an urban area of Brazil. Vet Parasitol 140: 54–60. doi: 10.1016/j.vetpar.2006.03.008
[47]
Mohebali M, Hajjaran H, Hamzavi Y, Mobedi I, Arshi S, et al. (2005) Epidemiological aspects of canine visceral leishmaniosis in the Islamic Republic of Iran. Vet Parasitol 129: 243–251. doi: 10.1016/j.vetpar.2005.01.010
[48]
Ertabaklar H, Toz SO, Ozkan AT, Rastgeldi S, Balcioglu IC, et al. (2005) Serological and entomological survey in a zoonotic visceral leishmaniasis focus of North Central Anatolia, Turkey: Corum province. Acta Trop 93: 239–246. doi: 10.1016/j.actatropica.2005.01.002
[49]
Molina R, Amela C, Nieto J, San-Andrés M, González F, et al. (1994) Infectivity of dogs naturally infected with Leishmania infantum to colonized Phlebotomus perniciosus. T Roy Soc Trop Med H 88: 491–493. doi: 10.1016/0035-9203(94)90446-4
[50]
Michalsky EM, Rocha MF, Da Rocha Lima AC, Fran?a-Silva JC, Pires MQ, et al. (2007) Infectivity of seropositive dogs, showing different clinical forms of leishmaniasis, to Lutzomyia longipalpis phlebotomine sand flies. Vet Parasitol 147: 67–76. doi: 10.1016/j.vetpar.2007.03.004
[51]
Alvar J, Molina R, San Andre ?S, Tesouro M, Nieto J, et al. (1994) Canine leishmaniasis: clinical, parasitological and entomological follow-up after chemotherapy. Ann Trop Med Parasit 88: 371–378.
[52]
Morillas F, Sanchez Rabasco F, Oca?a J, Martin-Sanchez J, Oca?a-Wihelmi J, et al. (1996) Leishmaniosis in the focus of the Axarquía region, Malaga province, southern Spain: a survey of the human, dog, and vector. Parasitol Res 82: 569–570. doi: 10.1007/s004360050164
[53]
Palatnik-de-Sousa CB, Dos Santos WR, Fran?a-Silva JC, da Costa RT, Reis AB, et al. (2001) Impact of canine control on the epidemiology of canine and human visceral leishmaniasis in Brazil. Am J Trop Med Hyg 65: 510–517.
[54]
Nasereddin A, Azmi K, Jaffe CL, Ereqat S, Amro A, et al. (2009) Kinetoplast DNA heterogeneity among Leishmania infantum strains in central Israel and Palestine. Vet Parasitol 161: 126–130. doi: 10.1016/j.vetpar.2008.12.003
[55]
Abdeen ZA, Sawalha SS, Eisenberger CL, Khanfar HM, Greenblatt CL, et al. (2002) Epidemiology of visceral leishmaniasis in the Jenin District, West Bank: 1989–1998. Am J Trop Med Hyg 66: 329–333.
[56]
Manna L, Vitale F, Reale S, Caracappa S, Pavone LM, et al. (2004) Comparison of different tissue sampling for PCR-based diagnosis and follow-up of canine visceral leishmaniosis. Vet Parasitol 125: 251–262. doi: 10.1016/j.vetpar.2004.07.019
[57]
Cascio A, Calattini S, Colomba C, Scalamogna C, Galazzi M, et al. (2002) Polymerase Chain Reaction in the Diagnosis and Prognosis of Mediterranean Visceral Leishmaniasis in Immunocompetent Children. Pediatrics 109: e27. doi: 10.1542/peds.109.2.e27
[58]
Noyes HA, Reyburn H, Bailey W, Smith D (1998) A nested-PCR-schizodeme method for identifying Leishmania kinetoplast minicircle classes directly from clinical samples and its application to the study of epidemiology of Leishmania tropica in Pakistan. J Clin Microbiol 36: 2877–2881.