Molecular Detection of Leishmania spp. in Skin and Blood of Stray Dogs from Endemic Areas of Cutaneous Leishmaniasis in Saudi Arabia
AbstractBackground: Dogs can act as reservoirs of canine leishmaniasis, caused by Leishmania species. The aims of this study were to determine the prevalence of canine leishmaniasis using a PCR technique among stray dogs living in three provinces of Saudi Arabia, Riyadh, Al-Ahsa Oasis and Al-Qaseem, where the disease is endemic; and to identify and document different Leishmania to species levels Methods: This cross-sectional investigation was conducted, from Mar 2016 to Apr 2018, in three parts of Saudi Arabia: Central province (Riyadh), Eastern province (Al-Ahsa Oasis) and Al-Qaseem province. Blood samples were collected from 526 dogs; 40 presented cutaneous nodules so were suspected clinically of cutaneous leishmaniasis. Biopsy tissue collections and parasite cultures were performed. A generic kDNA was performed using different primers for Leishmania differentiation. Results: All blood samples were negative for Leishmania infantum infection by molecular analysis, though forty dogs had thick cutaneous lesions in different parts of their body. Four dogs’ skin lesions were associated with dermatitis, splenomegaly and lymphadenomegaly. Parasite culture was used to diagnose cutaneous leishmaniasis, identifying 31/40 (77.5%) positive samples. Overall, of 526 samples, the prevalence of L. major and L. tropica was found to be 4% and 1.9%, respectively. Gender and age had a significant effect on Leishmania prevalence: (P=0.0212 and 0.0357), respectively. Conclusion: This was the first molecular study of dog leishmaniasis from Saudi Arabia of dogs confirmed to have cutaneous leishmaniasis. Further epidemiological and molecular investigations of domestic and wild canine infections with L. major, L. tropica and L. infantum in endemic and nonendemic areas of Saudi Arabia are required, for leishmaniasis control.
Ready PD. Epidemiology of visceral leishmaniasis. Clin Epidemiol 2014; 6:147.
Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den Boer M, Team WLC. Leishmaniasis worldwide and global estimates of its incidence. PloS one 2012; 7(5):e35671.
WHO. Sustaining the drive to overcome the global impact of neglected tropical diseases. Second Who Report on Neglected Tropical Diseases 2013;67-71.
Ashford R, Desjeux P. Estimation of population at risk of infection and number of cases of leishmaniasis. Parasitol Today 1992;8(3):104-105.
Marcili A, Sperança MA, da Costa AP, Madeira MdF, Soares HS, de OCC Sanches C, da CL Acosta I, Girotto A, Minervino AH, Horta MC. Phylogenetic relationships of Leishmania species based on trypanosomatid barcode (SSU rDNA) and gGAPDH genes: Taxonomic revision of Leishmania (L.) infantum chagasi in South America. Infect Genet Evol 2014; 25:44-51.
Shaw JJ. Further thoughts on the use of the name Leishmania (Leishmania) infantum chagasi for the aetiological agent of American visceral leishmaniasis. Mem Inst Oswaldo Cruz 2006; 101(5):577-579.
Al-Zahrani M, Peters W, Evans D, Smith V, Chin IC. Leishmania infecting man and wild animals in Saudi Arabia. 6. Cutaneous leishmaniasis of man in the south-west. Trans R Soc Trop Med Hyg 1989; 83(5):621-628.
Ibrahim E, Al-Zahrani M, Al-Tuwaigri A, Al-Shammary F, Evans D.Leishmania infecting man and wild animals in Saudi Arabia. 9. The black rat (Rattus rattus) a probable reservoir of visceral leishmaniasis in Gizan province, south-west Saudi Arabia. Trans R Soc Trop Med Hyg 1992; 86(5):513-514.
Al‐Qurashi AR, Ghandour AM, Osman M, Al‐Juma M. Dissemination in cutaneous leishmaniasis due to Leishmania major in different ethnic groups in Saudi Arabia. Int J Dermatol 2000;39(11):832-836.
Amin TT, Al-Mohammed HI, Kaliyadan F, Mohammed BS. Cutaneous leishmaniasis in Al Hassa, Saudi Arabia: epidemiological trends from 2000 to 2010. Asian Pac J Trop Dis 2013; 6(8):667-672.
Al-Tawfiq JA, AbuKhamsin A: Cutaneous leishmaniasis. a 46-year study of the epidemiology and clinical features in Saudi Arabia (1956–2002). T J Infect Dis 2004; 8(4):244-250.
Alanazi AD, Alyousif MS, Saifi MA, Alanazi IO. Epidemiological studies on cutaneous leishmaniasis in Ad-Dawadimi District, Saudi Arabia. Trop J Pharm Res 2016; 15(12):2709-2712.
Peters W, Elbihari S, Liu C, Le Blancq S, Evans D, Killick-Kendrick R, Smith V, Baldwin C.Leishmania infecting man and wild animals in Saudi Arabia 1. General survey. Trans R Soc Trop Med Hyg 1985; 79(6):831-839.
Elbihari S, Cheema AH, El-Hassan AM. Leishmania infecting man and wild animals in Saudi Arabia. 4. Canine cutaneous leishmaniasis in the Eastern Province. Trans R Soc Trop Med Hyg 1987; 81(6):925-927.
Silva RC, Richini-Pereira VB, Kikuti M, Marson PM, Langoni H. Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers. Vet Quart 2017; 37(1):23-30.
Salotra P, Sreenivas G, Pogue GP, Lee N, Nakhasi HL, Ramesh V, Negi N. Development of a species-specific PCR assay for detection of Leishmania donovani in clinical samples from patients with kala-azar and post-kala-azar dermal leishmaniasis. J Clin Microbiol 2001; 39(3):849-854.
Fichoux Y, Quaranta JF, Aufeuvre JP, Lelievre A, Marty P, Suffia I, Rousseau D, Kubar J. Occurrence of Leishmania infantum parasitemia in asymptomatic blood donors living in an area of endemicity in Southern France. J Clin Microbiol 1999; 37.
Hajjaran H, Mohebali M, Mamishi S, Vasigheh F, Oshaghi MA, Naddaf SR, Teimouri A, Edrissian GH, Zarei Z. Molecular identification and polymorphism determination of cutaneous and visceral leishmaniasis agents isolated from human and animal hosts in Iran. BioMed research international 2013;2013.
Abuzaid AA, Abdoon AM, Aldahan MA, Alzahrani AG, Alhakeem RF, Asiri AM, Alzahrani MH, Memish ZA. Cutaneous Leishmaniasis in Saudi Arabia: A Comprehensive Overview. Vector-Borne and Zoonotic Dis 2017;17(10):673-684.
Baneth G, Yasur-Landau D, Gilad M, Nachum-Biala Y. Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology. Parasites & Vectors 2017;10(1):113.
Mahboudi F, Abolhassani M, Yaran M, Mobtaker H, Azizi M. Identification and differentiation of Iranian Leishmania species by PCR amplification of kDNA. Scand J Infect Dis 2001; 33(8):596-598.
Cortes S, Rolão N, Ramada J, Campino L. PCR as a rapid and sensitive tool in the diagnosis of human and canine leishmaniasis using Leishmania donovani sl-specific kinetoplastid primers. Trans R Soc Trop Med Hyg 2004;98(1):12-17.
Bhutto AM, Soomro FR, Baloch JH, Matsumoto J, Uezato H, Hashiguchi Y, Katakura K. Cutaneous leishmaniasis caused by Leishmania (L.) major infection in Sindh province, Pakistan. Acta Tropica 2009;111(3):295-298.
Teles NMM, Agostini MAP, Bigeli JG, Noleto RV, Oliveira JD, Oliveira Junior WPd: Molecular and parasitological detection of Leishmania spp. in dogs caught in Palmas, TO, Brazil. Rev Bras Parasitol Vet 2012; 21(3):278-282.
Gao C-h, Ding D, Wang J-y, Steverding D, Wang X, Yang Y-t, Shi F. Development of a LAMP assay for detection of Leishmania infantum infection in dogs using conjunctival swab samples. Parasites & Vectors 2015; 8(1):370.
Bakirci S, Bilgiç HB, Köse O, Aksulu A, Hacilarlioğlu S, Erdoğan H, Karagenç T. Molecular and seroprevalence of canine visceral leishmaniasis in West Anatolia, Turkey. Turk J Vet Anim Sci. 2016, 40(5):637-644.
Ozerdem D, Eroglu F, Genc A, Demirkazik M, Koltas IS: Comparison of microscopic examination, rK39, and PCR for visceral leishmaniasis diagnosis in Turkey. Parasitol Res 2009; 106(1):197-200.
Anders G, Eisenberger CL, Jonas F, Greenblatt CL. Distinguishing Leishmania tropica and Leishmania major in the Middle East using the polymerase chain reaction with kinetoplast DNA-specific primers. Trans R Soc Trop Med Hyg 2002; 96:S87-S92.
Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol 2006;44(4):1435-1439.
Elbihari S, Kawasmeh Z, Al Naiem A: Possible reservoir host (s) of zoonotic cutaneous leishmaniasis in Al-Hassa oasis, Saudi Arabia. Ann Trop Med Parasitol 1984;78(5):543-545.
Morsy TA, Schnur LF, Feinsod FM, Salem AM, Wahba MM, El Said SM. Natural infections of Leishmania major in domestic dogs from Alexandria, Egypt. TAm J Trop Med Hyg 1987; 37(1):49-52.
Al-Bajalan MM, Niranji SS, Al-Jaf SM, Kato H: First identification of L. major in a dog in an endemic area of human cutaneous leishmaniasis in Iraq. molecular and phylogenetic studies. Parasitol Res 2017;1-6.
Baneth G, Nachum-Biala Y, Simon MS, Brenner O, Gaier S, Rojas A, Yasur-Landau D: Leishmania major infection in a dog with cutaneous manifestations. Parasites & Vectors 2016; 9(1):246.
Dereure J, Rioux J-A, Gallego M, Perieres J, Pratlong F, Mahjour J, Saddiki H.Leishmania tropica in Morocco: infection in dogs. Trans R Soc Trop Med Hyg 1991; 85(5):595.
Lemrani M, Nejjar R, Pratlong F: A new Leishmania tropica zymodeme--causative agent of canine visceral leishmaniasis in northern Morocco. Ann Trop Med Parasitol 2002; 96(6):637.
Bamorovat M, Sharifi I, Dabiri S, Mohammadi MA, Harandi MF, Mohebali M, Aflatoonian MR, Keyhani A. Leishmania tropica in stray dogs in southeast Iran. Iran J Public Health 2015; 44(10):1359.
Mohebali M, Malmasi A, Hajjaran H, Jamshidi S, Akhoundi B, Rezaei M, Janitabar S, Zarei H, Charehdar S: Disseminated leishmaniasis caused by Leishmania tropica in a puppy from Karaj, Central Iran. Iran J Parasitol 2011; 6(2):69.
Jacobson RL.Leishmaniasis in an era of conflict in the Middle East. Vector-Borne and Zoonotic Dis 2011; 11(3):247-258.
Colacicco-Mayhugh MG, Grieco JP, Putnam JL, Burkett DA, Coleman RE. Impact of phlebotomine sand flies on United States Military Operations at Tallil Air Base, Iraq: 5. Impact of weather on sand fly activity. J Med Entomol 2011;48(3):538-545.
Azizi K, Rassi Y, Javadian E, Motazedian M, Rafizadeh S, Yaghoobi Ershadi M, Mohebali M. Phlebotomus (Paraphlebotomus) alexandri: a probable vector of Leishmania infantum in Iran. Ann Trop Med Parasitol 2006; 100(1):63-68.
Dehkordi AS, Rassi Y, Oshaghi M, Abai M, Rafizadeh S, Yaghoobi-Ershadi M, Mohebali M, Zarei Z, Mohtarami F, Jafarzadeh B. Molecular detection of Leishmania infantum in naturally infected Phlebotomus perfiliewi transcaucasicus in Bilesavar district, northwestern Iran. Iran J Arthropod Borne Dis 2011; 5(1):20.
Svobodová M, Alten B, Zídková L, Dvořák V, Hlavačková J, Myšková J, Šeblová V, Kasap OE, Belen A, Votýpka J et al. Cutaneous leishmaniasis caused by Leishmania infantum transmitted by Phlebotomus tobbi. Int J Parasitol 2009; 39(2):251-256.
Baneth G, Zivotofsky D, Nachum-Biala Y, Yasur-Landau D, Botero A-M. Mucocutaneous Leishmania tropica infection in a dog from a human cutaneous leishmaniasis focus. Parasites & Vectors 2014; 7(1):118.