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Characterization of Leishmania isolates from two AIDS patients originating from Valencia, Spain
705 Kwok, S. & Higuchi, R. (1989). Avoiding false positives with PCR. Nature, 339,237-238. Manly, B. F. J. (1985). Appendix. In: The Statistics of Natural Selection on Animal Populations, Usher, M. B. & Rosenzweig, M. L. (editors). New York: Chapman & Hall, 424-431. Mantel, I. N. (1967). The detection of disease clustering and a generalized regression approach. Cancer Research, 27, 20% 220. Mazurier, S., Van De Giessen, A., Heuvelman, K. & Wernars, K. (1992). RAPD analysis of Campylobacter isolates: DNA tingerprinting without the need to purify DNA. Letters in AppliedMicrobiology, 14,260-262. McIntyre, P., Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1986). Chemotherapy in giardiasis: clinical responses and in vitro drug sensitivity of human isolates in axenic culture. ~ournalofPediatrics. 108.1005-1010. Meloni; B. I’. & Thompson, R. C. A. (1987). Corn arative studies on the axenic in vitro cultivation of Giardia o P human and canine origin: evidence for intraspecific variation. Transactions of the Royal Society of Tropical Medicine and Hygiene, 81,637-640. Meloni, B. I’., Lymbery, A. J. & Thompson, R. C. A. (1988). Isoenzyme electrophoresis of 30 isolates of Giardia from humans and felines. American Journal of Tropical Medicine and Hygiene, 38,65-73. Meloni, B. I’., Lymbery, A. J. & Thompson, R. C. A. (1989). Characterization of Gzardia isolates using a non-radiolabelled DNA probe, and correlation with the results of isoenzyme analysis. American Journal of Tropical Medicine and Hygiene,
40.629-637. --7
---
--.
Meloni, B. I’., Thompson, R. C. A., Stranden, A. M., Kohler, I’. & Eckert, I. (1992). Critical comuarison of Giardia duodenalis from Australia and Switzerland-using isoenzyme electrophoresis. Acta Tropica, 50, 115-124. Meyer, E. A. (1990). Preface. In: Giardiasis, Meyer, E. A. (editor). Amsterdam: Elsevier. Meyer, E. A. & Jarroll, E. J. (1980). Giardiasis. AmericanJournal of Epidemiology, lll, l-12. Nash, T. E. & Keister, D. B. (1985). Differences in excretorysecretory products and surface antigens among 19 isolates of
TRANSACTIONS OFTHE
ROYALSOCIETY OF TROPICAL
Giardia. Journal of InfectiousDiseases,152,1166-l 171. Nash, T .E., McCutchan, T., Keister, D., Dame, J. B., Conrad, I. D. & Gillin. F. D. (1985). Restriction-endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals.Joumal of InfectiousDiseases,152,64-73. Proctor, E. M., Isaac-Renton, J. L., Boyd, J., Wong, Q. At Bowie, W. R. (1989). Isoenzyme analysis of human and animal isolates of Giardia duodenalis from British Colombia, Canada. American Journal of Tropical Medicine and Hygiene, 41, 411-415. Smith, J. W. & Wolfe, M. S. (1980). Giardiasis. Annual Review of Medicine, 31,373-383. Sneath, P. H. A. (1957). Some thoughts on bacterial classification~oumal of General Microbiology, 17,201-226. Sneath, I’. H. A. & Sokal, R. R. (1973). Numerical Taxonomy. San Francisco: W. H. Freeman & Co. Thompson, R. C. A., Lymbery, A. J. & Meloni, B. I’. (1990). Genetic variation in Giardia Kunstler 1882: taxonomic and e$demiological significance. Protozoological Abstracts, 14, lThompson,
R. C. A., Reynoldson, J. A. & Mendis, A. H. W. and giardiasis. Advances in Parasitology, 32,
#W),Giardta
Welsh, J. & McClelland, M. (1991). Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18, 7213-7218. Welsh, J., Patersen, C. & McClelland, M. (1991). Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucleic Acids Research, 19,303-306. Williams, J. G. K., Kubehk, A. R., Livak, K. J., Rafalski, J. A. & Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18,6531-6535. Yu, K. & Pauls, K. I’. (1992). Optimization of the PCR program for RAPD analysis. Nucleic Acids Research, 20,2606.
Received 18 November 1992; revised 23 March accepted for publication 24 March 1993
1993;
MEDICINE AND HYGIENE (1993) 87, 705-706
Characterization of Leishmania isolates from two AIDS patients originating from Valencia, Spain F. Pratlongl, J. Reynes2, J. Dereurel, M. T. Baixenchs, B. Marchou4, M. Lefebvrel and F. Janbon2 ‘Laboratoire d’Ecologie Mddicale et Pathologie Parasitaire, Fact& de Medecine, Montpellier, France; 2Service des Maladies Infectieuses, Hopital Gui de Chauliac, 34059 Montpellier, France; 3Laboratoire de ParasitologieMycologic, Centre Hospitalier Universitaire, Hopital de Purpan, 31059 Toulouse, France; ?Service des Maladies Infectieuses, Hopital de Purpan, 310.59 Toulouse, France Introduction Visceral leishmaniasis (VL) associated with human immunodeficiency virus (HIV) infection has increased since 1985. Most of the reported cases originated in Mediterranean countries (ALTES et al., 1991; PETERS et al., 1990). The nrevalence of VL in nersons with HIV infection in souihern France was e&mated at between 2% (MARTY et al., 1989) in the Nice area and 3% in the Montpellier area (unpublished data). Only a few of the Leishmania isolates from HIV cases have been characterized by their isoenzyme profiles. The parasites which have been characterized .belong to the L. infantum complex, L. infantum zymodeme MON-1 being the most frequently encountered (MARTY et al., 1989).
This zymodeme is responsible for most VL in humans, dogs and foxes in the Mediterranean region. More recently, L. infantum zymodemes usually obtained from cutaneous leishmaniasis have been reported from VL associated with HIV infection (MON-24: GRADONI et al., 1990 and MARTY et al., 1991; MON-29: ALVAR et al., 1990; MON-33: CONDOM et al., 1989; MON-78: GRAMICCIA et al., 1992). A new L. infantum zymodeme (MON-136) was recently described in Italy from VL cases associated with HIV infection (GRAMICCIA et al., 1992). Here we report the isolation of a previously undescribed L. infantum zymodeme (MON-183) from associated infections of VL and HIV in 2 patients originating from the Valencia region, Spain. Case reports Case I. A 27 years old male intravenous drug user was first admitted to the infectious diseases department of the Centre Hospitalier Universitaire (CHU) in Montpellier, France, in July 1991 with fever of one month’s duration and weight loss (15 kg). Clinical examination revealed axillary adenopathies and splenomegaly. Results of enzyme-linked immunosorbent assay (ELISA) and Western blotting for antibody to HIV-l were positive and his blood contained 11 CD4 lvmnhocvtes/uL. Anaemia (haemoglobin=8.9g/dL) was assbciated with leucopenia (21OO/yL); the platelet count was 112 OOOiuL. A sternal puncture showed Leishmania amastigotes in smears, and NNN culture was positive (isolate MHOM/HES/91/ LEM 2298). By isoenzyme characterization, using 15 enzymatic systems (RIOUX et al., 1990), the isolate was 2
706
shown to be a new zymodeme (MON-183) of the L. infanztcmphenetic complex. This new zymodeme differed from the MON-1 zymodeme in 2 enzyme profiles (GP1115,MDHlo4). I mmunological reactions showed a hinh titre of fluorescent (IF11 antibodies (113201and negayive counter immunodlec&ophoresis (CIE). ‘Between July and August 1991 the patient received 2 courses of meglumine antimoniate (Glucantime@ ) (27.7g Sb total dose). In October 1991, the patient was readmitted with fever and hepatosplenomegaly. A bone marrow aspirate showed parasites and led to the cultivation of a new isolate (MHOM/HES/91/LEM 2361), again typed as L. infanhmz MON-183. Immunological results were similar to those obtained previously. A new course of Glucantime@ was iven (lOe2g Sb), followed by amphotericin B (Fungizone%+Intralipide@). The patient became apyrexic. Two further bone marrow aspirites were negative. -The patient died at the end of Ianuarv 1992 of cerebral toxonlasmosis. Postmortem exan&ation- showed Leishmania ody in the liver. Case2. A 28 years old male originating from Valencia, Spain, was admitted first to the neurological department and a few days later to the infectious diseasesdepartment of CHU, Purpan, Toulouse, France, in April 1991. He had been a drug abuser and was known to have been HIV seropositive since 1986. At hospital admission he presented fever, weight loss, asthenia and anorexia, but did not show any enlargement of liver or spleen. The blood picture showed pancytopenia, with haemoglobin 8.8 g/dL, leucocytes 15OO/pL,CD4 lymphocytes 16/yL and platelets 104 OOO/pL. The diagnosis of VL was based on the detection of numerous Leishmania amastigates in bone marrow aspirate (July 1991). The patient was treated with meglumine antimoniate (GlucantimP) for 5 d (3.1 e Sb total dose). But serious &de-effects led to the use ofalternative therapy with allopurinol (Zyloric@), a daily dose of 1500 mg for 15 d, followed by a daily dose of 500 mg for the following 30 d. After treatment, the clinical picture was satisfactory, in spite of persistance of an intermittent but well tolerated slight fever. In September 1991 the patient presented a leishmaniasis relapse with renewed fever, pancytopenia, and numerous Leishmania in the bone marrow. Treatment with intravenous pentamidine isethionate (Pentacarinat@)was given for imonths (100 mg twice dailv). Bone marrow culture on NNN medium led to the iso&ion of isolate MHOMIESI91ILBM10, which was characterized by isoenzyme electrophoresis as zymodeme MON-183. the sameas that from case1. Circulating antibodies w&e not detected by IF1 or CIE. The patient returned to Spain and was unavailable for follow-up. Discussion
endemic region (Toulouse), and frequently travelled to Valencia. VL is well known in the region of Valencia, but no systematic survey has been carried out and no isolate has been previously characterized. MON-183 is the first zymodeme isolated from this area. These observations emphasize the need for an eco-epidemiological study of this focus, from which the parasites have never been fully
characterized. Moreover, due to the increasing frequency of associations of leishmaniasis and acquired immune deficiency svndrome found in southern Europe, the characteriza&on of Leishmania isolates from patients with and without HIV infection is essential in order to evaluate the relationships between zymodemes and clinical expression. Acknowledgements We gratefully acknowledge Professor J. P. Dedet for his valuable discussion and for his help in the preparation of this manuscript. References AIt&, J., Salas, A., Riera, M., Udina, M., Galmes, A., Balanzat, J., Ballesteros, A., Buades, J., Salva, F. & Villalonga, C. (1991). Visceral leishmaniasis: another HIV-associated opportunistic infection? Report of eight casesand review of the literature. AIDS, 5,201-207. Alvar, J., Gutierrez-Solar, B., Benito, A., Aguiar? A., Cercenadq, E. & Barker, D. C. (1990). Characterizanon of Letshmanta stocks isolated from HIV positive patients. (VII Congr2sInternational de Parasitologic: Rbumts, abstract no. S3 Bl) Bulletin de la SocitW FranGuise de Parasitologic, 8, supolCment 1.232. Coidom, M.‘J., Clotet,, B.,, Sirera? G., Milla, F. & Foz, M. (1989). Asymptomauc lelshmamasis in the acquired immunodeficiency syndrome (AIDS). Annals of Internal Medicine, 111.767-768. ---‘: -’
--.
Gradom, L., Gramiccia, M. & Betti, F. (1990). Fatal visceral diseasecaused by a dermotropic Leishmania in a patient with human immunodeficiencv virus infection. Youmal of_ Znfec_ tion, 20, 180-182. . Gramiccia, M., Gradoni, L. & Troiani, M. (1992). HIV-Leishmania co-infections in Italv. Isoenzvme characterization of Leishmania causing viscerai leishmaiiasis in HIV patients. Transactions of the Royal Sociely of Tropical Medicine and Hygiene, 86,161-163. Marty, P., Pesce,A., Fuzibet, J. G., Bernard, E., Bertrand, F., Saint-Paul, M. C., Gari-Toussaint, M. & Le Fichoux, Y. (1989). Aspects biocliniques de la leishmaniose viscerale chez les sideens. A propos de 7 observations au C.H.U. de Nice. Bulletin de la Sock% FranGaise de Parasitologie, 7, 159161. Marty, I’., Fuzibet, J. G., Pratlong, F., Quinsat, D., GariToussaint, M., Dor, J. F., Le Fichoux, Y. & Rioux, J. A. (1991). Leishmaniose viscerale causee par une souche dermotrope de Leishmania infantum chez un sideen. Bulletin de la Sock% de Pathologie Exotique, 84,365-367. Peters, B. S., Fish, D., Golden, R., Evans, D. A., Bryceson, A. D. M. & Pinching, A. J. (1990). Visceral leishmaniasis in HIV infection and AIDS: clinical features and response to therapy. QuarterlyJournal of Medicine, 283,110 l-l 111. Rioux, J. A., Moreno, G., Lanotte, G., Pratlong, F., Dereure, J. & Rispail, P. (1986). Two episodes of cutaneous leishmaniasis in man caused by different zymodemes of Leishmania
The new L. infanturn zymodeme MON-183 was isolated from 2 patients whose infection had presumably occurred in the sameregion of Spain (Valencia). The first patient lived near Montpellier, in a leishmaniasis focus where only L. infanturn zymodemes MON-1 and MON-29 had been found previously (RIOUX et al., 1986), but he had travelled to the Valencia region of Spain during the summer transmission seasonof 1990. The second patient originated from the same area of
Received 4 Januay
Spain. He had lived in France only since 1990, in a non-
for publication 4 March 1993
infantum s.1. Transactions of the Royal Society of Tropical Medicine and Hveiene. 80.1004-1005.
Rioux, J. A., LanGfie, 6.) gerres, E., Pratlong, F., Bastien, P. & P&i&es, J. (1990). Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Annales de Parasitologie Humaine et Cornparke, 65, 111-125. 1993; revised I March 1993; accepted
40.629-637. --7
---
--.
Meloni, B. I’., Thompson, R. C. A., Stranden, A. M., Kohler, I’. & Eckert, I. (1992). Critical comuarison of Giardia duodenalis from Australia and Switzerland-using isoenzyme electrophoresis. Acta Tropica, 50, 115-124. Meyer, E. A. (1990). Preface. In: Giardiasis, Meyer, E. A. (editor). Amsterdam: Elsevier. Meyer, E. A. & Jarroll, E. J. (1980). Giardiasis. AmericanJournal of Epidemiology, lll, l-12. Nash, T. E. & Keister, D. B. (1985). Differences in excretorysecretory products and surface antigens among 19 isolates of
TRANSACTIONS OFTHE
ROYALSOCIETY OF TROPICAL
Giardia. Journal of InfectiousDiseases,152,1166-l 171. Nash, T .E., McCutchan, T., Keister, D., Dame, J. B., Conrad, I. D. & Gillin. F. D. (1985). Restriction-endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals.Joumal of InfectiousDiseases,152,64-73. Proctor, E. M., Isaac-Renton, J. L., Boyd, J., Wong, Q. At Bowie, W. R. (1989). Isoenzyme analysis of human and animal isolates of Giardia duodenalis from British Colombia, Canada. American Journal of Tropical Medicine and Hygiene, 41, 411-415. Smith, J. W. & Wolfe, M. S. (1980). Giardiasis. Annual Review of Medicine, 31,373-383. Sneath, P. H. A. (1957). Some thoughts on bacterial classification~oumal of General Microbiology, 17,201-226. Sneath, I’. H. A. & Sokal, R. R. (1973). Numerical Taxonomy. San Francisco: W. H. Freeman & Co. Thompson, R. C. A., Lymbery, A. J. & Meloni, B. I’. (1990). Genetic variation in Giardia Kunstler 1882: taxonomic and e$demiological significance. Protozoological Abstracts, 14, lThompson,
R. C. A., Reynoldson, J. A. & Mendis, A. H. W. and giardiasis. Advances in Parasitology, 32,
#W),Giardta
Welsh, J. & McClelland, M. (1991). Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18, 7213-7218. Welsh, J., Patersen, C. & McClelland, M. (1991). Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucleic Acids Research, 19,303-306. Williams, J. G. K., Kubehk, A. R., Livak, K. J., Rafalski, J. A. & Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18,6531-6535. Yu, K. & Pauls, K. I’. (1992). Optimization of the PCR program for RAPD analysis. Nucleic Acids Research, 20,2606.
Received 18 November 1992; revised 23 March accepted for publication 24 March 1993
1993;
MEDICINE AND HYGIENE (1993) 87, 705-706
Characterization of Leishmania isolates from two AIDS patients originating from Valencia, Spain F. Pratlongl, J. Reynes2, J. Dereurel, M. T. Baixenchs, B. Marchou4, M. Lefebvrel and F. Janbon2 ‘Laboratoire d’Ecologie Mddicale et Pathologie Parasitaire, Fact& de Medecine, Montpellier, France; 2Service des Maladies Infectieuses, Hopital Gui de Chauliac, 34059 Montpellier, France; 3Laboratoire de ParasitologieMycologic, Centre Hospitalier Universitaire, Hopital de Purpan, 31059 Toulouse, France; ?Service des Maladies Infectieuses, Hopital de Purpan, 310.59 Toulouse, France Introduction Visceral leishmaniasis (VL) associated with human immunodeficiency virus (HIV) infection has increased since 1985. Most of the reported cases originated in Mediterranean countries (ALTES et al., 1991; PETERS et al., 1990). The nrevalence of VL in nersons with HIV infection in souihern France was e&mated at between 2% (MARTY et al., 1989) in the Nice area and 3% in the Montpellier area (unpublished data). Only a few of the Leishmania isolates from HIV cases have been characterized by their isoenzyme profiles. The parasites which have been characterized .belong to the L. infantum complex, L. infantum zymodeme MON-1 being the most frequently encountered (MARTY et al., 1989).
This zymodeme is responsible for most VL in humans, dogs and foxes in the Mediterranean region. More recently, L. infantum zymodemes usually obtained from cutaneous leishmaniasis have been reported from VL associated with HIV infection (MON-24: GRADONI et al., 1990 and MARTY et al., 1991; MON-29: ALVAR et al., 1990; MON-33: CONDOM et al., 1989; MON-78: GRAMICCIA et al., 1992). A new L. infantum zymodeme (MON-136) was recently described in Italy from VL cases associated with HIV infection (GRAMICCIA et al., 1992). Here we report the isolation of a previously undescribed L. infantum zymodeme (MON-183) from associated infections of VL and HIV in 2 patients originating from the Valencia region, Spain. Case reports Case I. A 27 years old male intravenous drug user was first admitted to the infectious diseases department of the Centre Hospitalier Universitaire (CHU) in Montpellier, France, in July 1991 with fever of one month’s duration and weight loss (15 kg). Clinical examination revealed axillary adenopathies and splenomegaly. Results of enzyme-linked immunosorbent assay (ELISA) and Western blotting for antibody to HIV-l were positive and his blood contained 11 CD4 lvmnhocvtes/uL. Anaemia (haemoglobin=8.9g/dL) was assbciated with leucopenia (21OO/yL); the platelet count was 112 OOOiuL. A sternal puncture showed Leishmania amastigotes in smears, and NNN culture was positive (isolate MHOM/HES/91/ LEM 2298). By isoenzyme characterization, using 15 enzymatic systems (RIOUX et al., 1990), the isolate was 2
706
shown to be a new zymodeme (MON-183) of the L. infanztcmphenetic complex. This new zymodeme differed from the MON-1 zymodeme in 2 enzyme profiles (GP1115,MDHlo4). I mmunological reactions showed a hinh titre of fluorescent (IF11 antibodies (113201and negayive counter immunodlec&ophoresis (CIE). ‘Between July and August 1991 the patient received 2 courses of meglumine antimoniate (Glucantime@ ) (27.7g Sb total dose). In October 1991, the patient was readmitted with fever and hepatosplenomegaly. A bone marrow aspirate showed parasites and led to the cultivation of a new isolate (MHOM/HES/91/LEM 2361), again typed as L. infanhmz MON-183. Immunological results were similar to those obtained previously. A new course of Glucantime@ was iven (lOe2g Sb), followed by amphotericin B (Fungizone%+Intralipide@). The patient became apyrexic. Two further bone marrow aspirites were negative. -The patient died at the end of Ianuarv 1992 of cerebral toxonlasmosis. Postmortem exan&ation- showed Leishmania ody in the liver. Case2. A 28 years old male originating from Valencia, Spain, was admitted first to the neurological department and a few days later to the infectious diseasesdepartment of CHU, Purpan, Toulouse, France, in April 1991. He had been a drug abuser and was known to have been HIV seropositive since 1986. At hospital admission he presented fever, weight loss, asthenia and anorexia, but did not show any enlargement of liver or spleen. The blood picture showed pancytopenia, with haemoglobin 8.8 g/dL, leucocytes 15OO/pL,CD4 lymphocytes 16/yL and platelets 104 OOO/pL. The diagnosis of VL was based on the detection of numerous Leishmania amastigates in bone marrow aspirate (July 1991). The patient was treated with meglumine antimoniate (GlucantimP) for 5 d (3.1 e Sb total dose). But serious &de-effects led to the use ofalternative therapy with allopurinol (Zyloric@), a daily dose of 1500 mg for 15 d, followed by a daily dose of 500 mg for the following 30 d. After treatment, the clinical picture was satisfactory, in spite of persistance of an intermittent but well tolerated slight fever. In September 1991 the patient presented a leishmaniasis relapse with renewed fever, pancytopenia, and numerous Leishmania in the bone marrow. Treatment with intravenous pentamidine isethionate (Pentacarinat@)was given for imonths (100 mg twice dailv). Bone marrow culture on NNN medium led to the iso&ion of isolate MHOMIESI91ILBM10, which was characterized by isoenzyme electrophoresis as zymodeme MON-183. the sameas that from case1. Circulating antibodies w&e not detected by IF1 or CIE. The patient returned to Spain and was unavailable for follow-up. Discussion
endemic region (Toulouse), and frequently travelled to Valencia. VL is well known in the region of Valencia, but no systematic survey has been carried out and no isolate has been previously characterized. MON-183 is the first zymodeme isolated from this area. These observations emphasize the need for an eco-epidemiological study of this focus, from which the parasites have never been fully
characterized. Moreover, due to the increasing frequency of associations of leishmaniasis and acquired immune deficiency svndrome found in southern Europe, the characteriza&on of Leishmania isolates from patients with and without HIV infection is essential in order to evaluate the relationships between zymodemes and clinical expression. Acknowledgements We gratefully acknowledge Professor J. P. Dedet for his valuable discussion and for his help in the preparation of this manuscript. References AIt&, J., Salas, A., Riera, M., Udina, M., Galmes, A., Balanzat, J., Ballesteros, A., Buades, J., Salva, F. & Villalonga, C. (1991). Visceral leishmaniasis: another HIV-associated opportunistic infection? Report of eight casesand review of the literature. AIDS, 5,201-207. Alvar, J., Gutierrez-Solar, B., Benito, A., Aguiar? A., Cercenadq, E. & Barker, D. C. (1990). Characterizanon of Letshmanta stocks isolated from HIV positive patients. (VII Congr2sInternational de Parasitologic: Rbumts, abstract no. S3 Bl) Bulletin de la SocitW FranGuise de Parasitologic, 8, supolCment 1.232. Coidom, M.‘J., Clotet,, B.,, Sirera? G., Milla, F. & Foz, M. (1989). Asymptomauc lelshmamasis in the acquired immunodeficiency syndrome (AIDS). Annals of Internal Medicine, 111.767-768. ---‘: -’
--.
Gradom, L., Gramiccia, M. & Betti, F. (1990). Fatal visceral diseasecaused by a dermotropic Leishmania in a patient with human immunodeficiencv virus infection. Youmal of_ Znfec_ tion, 20, 180-182. . Gramiccia, M., Gradoni, L. & Troiani, M. (1992). HIV-Leishmania co-infections in Italv. Isoenzvme characterization of Leishmania causing viscerai leishmaiiasis in HIV patients. Transactions of the Royal Sociely of Tropical Medicine and Hygiene, 86,161-163. Marty, P., Pesce,A., Fuzibet, J. G., Bernard, E., Bertrand, F., Saint-Paul, M. C., Gari-Toussaint, M. & Le Fichoux, Y. (1989). Aspects biocliniques de la leishmaniose viscerale chez les sideens. A propos de 7 observations au C.H.U. de Nice. Bulletin de la Sock% FranGaise de Parasitologie, 7, 159161. Marty, I’., Fuzibet, J. G., Pratlong, F., Quinsat, D., GariToussaint, M., Dor, J. F., Le Fichoux, Y. & Rioux, J. A. (1991). Leishmaniose viscerale causee par une souche dermotrope de Leishmania infantum chez un sideen. Bulletin de la Sock% de Pathologie Exotique, 84,365-367. Peters, B. S., Fish, D., Golden, R., Evans, D. A., Bryceson, A. D. M. & Pinching, A. J. (1990). Visceral leishmaniasis in HIV infection and AIDS: clinical features and response to therapy. QuarterlyJournal of Medicine, 283,110 l-l 111. Rioux, J. A., Moreno, G., Lanotte, G., Pratlong, F., Dereure, J. & Rispail, P. (1986). Two episodes of cutaneous leishmaniasis in man caused by different zymodemes of Leishmania
The new L. infanturn zymodeme MON-183 was isolated from 2 patients whose infection had presumably occurred in the sameregion of Spain (Valencia). The first patient lived near Montpellier, in a leishmaniasis focus where only L. infanturn zymodemes MON-1 and MON-29 had been found previously (RIOUX et al., 1986), but he had travelled to the Valencia region of Spain during the summer transmission seasonof 1990. The second patient originated from the same area of
Received 4 Januay
Spain. He had lived in France only since 1990, in a non-
for publication 4 March 1993
infantum s.1. Transactions of the Royal Society of Tropical Medicine and Hveiene. 80.1004-1005.
Rioux, J. A., LanGfie, 6.) gerres, E., Pratlong, F., Bastien, P. & P&i&es, J. (1990). Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Annales de Parasitologie Humaine et Cornparke, 65, 111-125. 1993; revised I March 1993; accepted