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The characterization of Leishmania major from Phlebotomus papatasi (Scopoli) caught in northern Sinai, Egypt
TRANSA,~IONS OF THE ROYALSca~rv OF TROPICALMEDICINEAND HYGIENE(1990)
785
84, 785-786
1 Short Report 1 The characterization of Leishmania majorfrom Phlebotomus papatasi (Scopoli) caught in northern Sinai, Egypt M. M. Wahba’, L. F. Schnu?, T. A. Morsy’ and A. ‘Ain Shams Research and Training Centre Me&n’ on Vectors of Diseases, Ain Shams University, Abassia, Cairo, Egypt; 2Kuvin Centre for the Study of Infectious and Tropical Diseases, Hebrew University-Hadassah Medical School, Jerusalem, Israel Introduction
The Sinai Peninsula, Egypt’s eastern province, is bordered by countries where cutaneous leishmaniasis has been recorded (SCHLEINet al., 1984; PETERSet al., 1985). During 1988 and 1989, 17 human casesof cutaneous leishmaniasis (CL) were recorded by local health services. Also, wild rodents of the genera Gerbillus, Meriones and Jaculus abound (OSBORN & HELMY, 1980). The parasites causing CL in Gerbillus pyramidurn (MORW et al., 1987) and man (MANSOUR et al., 198!; GREENBLATTet al., 1990) have been typed asLewhmania major. Phlebotomus papa& is the commonest species of sandfly in northern Sinai (EL SAWAFet al., 1987). Here, we present the results of a two-year investigation on leishmanial infections in sandflies. Materials
and Methods
Sandflies were collected in different localities of northern Sinai during the months of Mav to November in 1988and 1989rCDC miniature light traps were placed in human dwellings. Flies were kept alive and dissected in our field laboratory, according to the method described by R~OUX et al. (1984). Female sandflies were identified by pharynged &ature and swrmathecal mornholoav (LANE 1986). About 4000 fimale P. papatai were%&.mined for ihe presenceof promastigotes; 6 were found to be infected. These 6 were from a batch collected on 1 November 1989, at Sad El Rawafai, 60 km south-east of El Arish in northern Sinai. Parasites were isolated in NNN medium. The leishmanial stock isolated and subsequently typed, IPAP/EG/89IRTC-13, was grown in brain-heart infusion agar and characterized by enzyme electrophoresis, comparing it with international reference s&ins supplied by tile World Health Organization Jerusalem Reference Centre for the Leishmaniases: MRHO/SU/SB/P and MHOM/IL/67/Jericho II (=LRC-L137) for L. major, MHOM/TNWO/IPTl for L. infantum, and MHOMBQ165IASI (=LRC-L32) for L. tropica. Enzyme analysis was by cellulose acetate electrophoresis (KREUTZERet al., 1987; EVANSet al., 1989). Twelve enzymes were examined: malate dehydrogenase (MDH, EC. 1.1, 1.37), phosphogluconate dehydrogenase (6PGD, EC. 1.1.1.44), phosphoglucomutase (PGM, EC.2.7.5. l), superoxide dismutase (SOD, EC.l.lS.l.l.), aspartate aminotrans-
ferase(ASAT, EC.2.6.1.1.), alanine aminotransferase (ALAT, EC.2.6.1.2.), pyruvate kinase (PK, EC.2.7.40), nucleoside hydrolase (NH, EC.3.2.2.2.), proline immopeptidase (PEPD, EC.3.4.11.5.), mannose phosphate isomerase (MPI, EC.5.3.1.8.), glucose phosphate isomerase (GPI, EC.5.3.1.9.>, and malic enzyme (ME, EC. 1.1.1.40.). Results
The stock was identical to the international reference strain MHOM/IU67/Jericho II (=LRC-L37), which belongs to zymodeme LON70 of L. major (D. A. Evans, personal communication) and to a local reference strain, MHOM/EG/89/RTC-10, isolated from a human cutaneous case. Conclusion
The characterization of strain RTC-13 is the first definite evidence incriminating P. papatasi as the vector of L. major and human CL in northern Sinai, fulfilling the criteria listed by KILLICK-KENDRICK i? WARD (1981) and KILLICK-KENDRICK
(1990).
Acknowledgements
This work was supported by the Regional Project ‘Epidemiology and Control of Arthropod-Borne Diseasesin Egypt’, NIH-NIAID contract NOI AI 22667. We thank Mr Said Doha for help.
References
El Sawaf, B. M., Shoukry, A., El Said, S., Lane, R. P., Kenawy, M. A., Beier, J. C. & Abdel Sattar, S. (1987). Sandfly speciescomposition along an altitudinal transect in southern Sinai, Egypt. Andes de Parasitologic Humaine et Compart!e, 62, 467473.
Evans, D., Godfrey, D., Lanham, S., Lanotte, G., Modabber, F. & Schnur, L. (1989). Handbook on Isolation, Characterization and Cyopresefvation of Leishmania. Geneva: UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases. Greenblatt, C. L., Schnur, L. F., Juster, R. & Sulitzeanu, ;4js(1990). Israel Journal of Medical Sciences,26, 129Killick-Kendrick, R. (1990). Phlebomine vectors of the leishmaniases:a review. Medical and Veterinary Entomology, 4, l-24. Killick-Kendrick, R. & Ward, R. D. (1981). The ecology of Leishmania. Parasitology, 82, 143-152. Kreutzer, R. D., Souraty, N. & Semko, M. (1987). Biochemical identities and differences among Leishmania species and subspecies. American 3ouwuiof Tropical Medicine and Hygiene, 36, 22-32.
Lane, R. P. (1986). The sandflies of Egypt (Diptera: Phlebotominae). Bulletin of the British Museum (National History), (Entomology), 52, l-35. Mansour, N. S., Youssef, F. G., Mohareb, E. W., Dees, W. H. & Karuru, E. R. (1987). Cutaneous leishmaniasis in north Sinai. Transactions of the Royal Society of Tropical Medicine and Hygiene, 81, 747.
Morsy, T. A., Shoukry,A., Schnur,L. F. & Sulitzeanu, A.
(1987). Gerbillus pyramidum is a host of Leishmania major in the Sinai Peninsula. Annals of Tropical Medicine and
Parasitolagy,
81, 741-742.
Osbom, D. J. & Helm?, I. (1980). The Contemporary Land Mammals of Egypt (m&ding Sinal]. Fieldiana: Zoology, new series no. 5. The Field Museum of Natural History, publication no. 1309.
786 Ckvennes.Annales deParasitologic Humaine et Comparde,
Peters, W., Elbihari, S., Ching Lui, Le Blancq, S. M.
5q, 331-333.
Evans?D. A., Killick-Kendrick, R., Smith, V. & Baldwm,C. I. (1985).Z.eis~mania infectingmanand wild animalsin SaudiArabia. General survey. Transactions of
Schlem, Y., Warburg,A., Schnur,L. F., Le Blancq,S. M. & Gunders, A. E. (1984). Leishmaniasis in Israel:
the Royal Society of Tropical Medicine and Hygiene, 79,
reservoirhosts,sandtlyvectorsandleishmanialstrainsin the Negev, Central Arava and along the Dead Sea.
831-839. Rioux, J.-A., Jarry, D. M., Lanotte, G., Maazoun,R. & Killick-Kendrick, R. (1984).Ecologicdesleishmanioses dam le sud de la France.18Identification enzymatique de L&.&mania infantum Nicolle, 1908, isolC de Phleboto-
j&;ived
TRANSACTIONSOF THE ROYAL SOCIETYOF TROPICALMEDICINE
HYGIENE(1990) 84, 786
mus ariasi Tonnoir,
1921, spontanement infest6 en
1 Short Report 1 The importance of rapid diagnosis of new cases of cutaneous leishmaniasis in pin-pointing the sandfly vector L. Ryad*, A. Vexenag, P. D. Marsden’, R. Lains~d +d J. J. Shawl ‘The Wellcome Parasitolp p Umt, Carxa Postal 3, t$5*001, Belhn, Par& Bpztl; Ntileo de Medicina Troprcal e Nutripio, Umverstdade de Bra&a, Brash, DF, Brazil
We have searched for many years for the vector of Leiskmania (Viunnia) braziliensis in the endemic area
of T&s Bra$os, Bahia, Brazil. Since over 95.1% of the phlebotomine santies captured in and around houses are Lutzomyia wkitmani (Antunes & Coutinho), and leishmanial ulcers were occasionally seenin &all, house-bound children, this specieswas susnected to be the vector WEXENAT et al., 1986). $&al efforts were made t&dissect sandf3iescaught around the houses of families where more than one caseof acute cutaneous leishmaniasis was present and, although several such attempts failed; we briefli record here the lirst successful isolation of L. (V.) braziliensis from Lu. wkitmuni as a result of this policy. In January, 1986 two men attended the field clinic, both with ulceration of the lower leg of 15 d duration. They were accustomed to meet in the evening, frequently clad in shorts and thong sandles. They discussed the day’s events while seated on a log in front of their adjoining houses in Fazenda Rizada, a
AND
Transactions of the Royal Society of Tropical Medicine and Hygiene, 78, 48U84. 21 May 1990; accepted
12July
farm currently under longitudinal study and where 6 casesof cutaneous leishmaniasis have been recorded during the decade 1980-1990. Parasites later identiiied as L. (V.) braziliensis were isolated from both patients (LTB:865 and LTB:866) and treatment with Glucantime commenced. Two days later entomological studies were initiated and concentrated on this locality: in the next few days 193Lu. wkimni were captured from a chicken-house near the house of one of the patients and, on dissection, one was found to be inf&ted with L..(V.) braziltis (RYAN et al.. 1987). Identification was made in bdth the Be& and ‘Brasilia laboratories, using monoclonal antibodies and enzyme profiles (strain IWHIIBR/86/M10187). The very recent nature of the human infections undoubtedly helped in our selection of an appropriate sandfly capture-site. Usually patients arrive for consultation later in the disease,but on this occasion the farm was near the medical post and the men had a car at their disposal. Later in the sameyear (HOCK et al., 1986) 2 further isolates of L. (V.) braziliensis were made from Lu. whitmani, on a farm where several children were infected in the same family. References
Hoch, A., Ryan, L., Vexenat, J. A., Rosa, A. C. 0. C. & Barreto. A. C. (1986). Isolation of Leishmania brazilknsis braziliekis
and‘o&r trypanosomatidsfrom phleboto-
mines in a mucocutaneousleishmaniasis endemic area in Bahia, Brazil. Mem6rias do Inszinrto Oswald0 Cruz, 81, supplkment?abstract no. Bl 44. Ryan, L., Lamson, R., Shaw, J. J., Braga, R. R. 81 Ishikawa, E. A. Y. (1987). Leishmaniasis in Brazil. XXV. Sandfly vectors of Leishmania in Par6 State, Brazil. Medical and Veterinary Enmwwlogy, 1, 383-395. Vexenat, J. A., Barreto, A. C., Cuba, C. C. & Marsden,P. D. (1986). Caracteristicas epidemioldgicas da leishmaniose tegumentar americana em uma regiao end&mica do Estado da Bahia. III. Fauna flebotomlnica. MenGrius do Institute Oswald0 Crw,
*Present address: Philip Morris USA ResearchCentre, P.O. Box 26583, Richmond, VA 23261, USA. Off-print requests to R. Lainson.
for publicatkm
Received
1990
l2Jun.t~
81, 293-301.
1990; acceptedfmpublication
ZJune
785
84, 785-786
1 Short Report 1 The characterization of Leishmania majorfrom Phlebotomus papatasi (Scopoli) caught in northern Sinai, Egypt M. M. Wahba’, L. F. Schnu?, T. A. Morsy’ and A. ‘Ain Shams Research and Training Centre Me&n’ on Vectors of Diseases, Ain Shams University, Abassia, Cairo, Egypt; 2Kuvin Centre for the Study of Infectious and Tropical Diseases, Hebrew University-Hadassah Medical School, Jerusalem, Israel Introduction
The Sinai Peninsula, Egypt’s eastern province, is bordered by countries where cutaneous leishmaniasis has been recorded (SCHLEINet al., 1984; PETERSet al., 1985). During 1988 and 1989, 17 human casesof cutaneous leishmaniasis (CL) were recorded by local health services. Also, wild rodents of the genera Gerbillus, Meriones and Jaculus abound (OSBORN & HELMY, 1980). The parasites causing CL in Gerbillus pyramidurn (MORW et al., 1987) and man (MANSOUR et al., 198!; GREENBLATTet al., 1990) have been typed asLewhmania major. Phlebotomus papa& is the commonest species of sandfly in northern Sinai (EL SAWAFet al., 1987). Here, we present the results of a two-year investigation on leishmanial infections in sandflies. Materials
and Methods
Sandflies were collected in different localities of northern Sinai during the months of Mav to November in 1988and 1989rCDC miniature light traps were placed in human dwellings. Flies were kept alive and dissected in our field laboratory, according to the method described by R~OUX et al. (1984). Female sandflies were identified by pharynged &ature and swrmathecal mornholoav (LANE 1986). About 4000 fimale P. papatai were%&.mined for ihe presenceof promastigotes; 6 were found to be infected. These 6 were from a batch collected on 1 November 1989, at Sad El Rawafai, 60 km south-east of El Arish in northern Sinai. Parasites were isolated in NNN medium. The leishmanial stock isolated and subsequently typed, IPAP/EG/89IRTC-13, was grown in brain-heart infusion agar and characterized by enzyme electrophoresis, comparing it with international reference s&ins supplied by tile World Health Organization Jerusalem Reference Centre for the Leishmaniases: MRHO/SU/SB/P and MHOM/IL/67/Jericho II (=LRC-L137) for L. major, MHOM/TNWO/IPTl for L. infantum, and MHOMBQ165IASI (=LRC-L32) for L. tropica. Enzyme analysis was by cellulose acetate electrophoresis (KREUTZERet al., 1987; EVANSet al., 1989). Twelve enzymes were examined: malate dehydrogenase (MDH, EC. 1.1, 1.37), phosphogluconate dehydrogenase (6PGD, EC. 1.1.1.44), phosphoglucomutase (PGM, EC.2.7.5. l), superoxide dismutase (SOD, EC.l.lS.l.l.), aspartate aminotrans-
ferase(ASAT, EC.2.6.1.1.), alanine aminotransferase (ALAT, EC.2.6.1.2.), pyruvate kinase (PK, EC.2.7.40), nucleoside hydrolase (NH, EC.3.2.2.2.), proline immopeptidase (PEPD, EC.3.4.11.5.), mannose phosphate isomerase (MPI, EC.5.3.1.8.), glucose phosphate isomerase (GPI, EC.5.3.1.9.>, and malic enzyme (ME, EC. 1.1.1.40.). Results
The stock was identical to the international reference strain MHOM/IU67/Jericho II (=LRC-L37), which belongs to zymodeme LON70 of L. major (D. A. Evans, personal communication) and to a local reference strain, MHOM/EG/89/RTC-10, isolated from a human cutaneous case. Conclusion
The characterization of strain RTC-13 is the first definite evidence incriminating P. papatasi as the vector of L. major and human CL in northern Sinai, fulfilling the criteria listed by KILLICK-KENDRICK i? WARD (1981) and KILLICK-KENDRICK
(1990).
Acknowledgements
This work was supported by the Regional Project ‘Epidemiology and Control of Arthropod-Borne Diseasesin Egypt’, NIH-NIAID contract NOI AI 22667. We thank Mr Said Doha for help.
References
El Sawaf, B. M., Shoukry, A., El Said, S., Lane, R. P., Kenawy, M. A., Beier, J. C. & Abdel Sattar, S. (1987). Sandfly speciescomposition along an altitudinal transect in southern Sinai, Egypt. Andes de Parasitologic Humaine et Compart!e, 62, 467473.
Evans, D., Godfrey, D., Lanham, S., Lanotte, G., Modabber, F. & Schnur, L. (1989). Handbook on Isolation, Characterization and Cyopresefvation of Leishmania. Geneva: UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases. Greenblatt, C. L., Schnur, L. F., Juster, R. & Sulitzeanu, ;4js(1990). Israel Journal of Medical Sciences,26, 129Killick-Kendrick, R. (1990). Phlebomine vectors of the leishmaniases:a review. Medical and Veterinary Entomology, 4, l-24. Killick-Kendrick, R. & Ward, R. D. (1981). The ecology of Leishmania. Parasitology, 82, 143-152. Kreutzer, R. D., Souraty, N. & Semko, M. (1987). Biochemical identities and differences among Leishmania species and subspecies. American 3ouwuiof Tropical Medicine and Hygiene, 36, 22-32.
Lane, R. P. (1986). The sandflies of Egypt (Diptera: Phlebotominae). Bulletin of the British Museum (National History), (Entomology), 52, l-35. Mansour, N. S., Youssef, F. G., Mohareb, E. W., Dees, W. H. & Karuru, E. R. (1987). Cutaneous leishmaniasis in north Sinai. Transactions of the Royal Society of Tropical Medicine and Hygiene, 81, 747.
Morsy, T. A., Shoukry,A., Schnur,L. F. & Sulitzeanu, A.
(1987). Gerbillus pyramidum is a host of Leishmania major in the Sinai Peninsula. Annals of Tropical Medicine and
Parasitolagy,
81, 741-742.
Osbom, D. J. & Helm?, I. (1980). The Contemporary Land Mammals of Egypt (m&ding Sinal]. Fieldiana: Zoology, new series no. 5. The Field Museum of Natural History, publication no. 1309.
786 Ckvennes.Annales deParasitologic Humaine et Comparde,
Peters, W., Elbihari, S., Ching Lui, Le Blancq, S. M.
5q, 331-333.
Evans?D. A., Killick-Kendrick, R., Smith, V. & Baldwm,C. I. (1985).Z.eis~mania infectingmanand wild animalsin SaudiArabia. General survey. Transactions of
Schlem, Y., Warburg,A., Schnur,L. F., Le Blancq,S. M. & Gunders, A. E. (1984). Leishmaniasis in Israel:
the Royal Society of Tropical Medicine and Hygiene, 79,
reservoirhosts,sandtlyvectorsandleishmanialstrainsin the Negev, Central Arava and along the Dead Sea.
831-839. Rioux, J.-A., Jarry, D. M., Lanotte, G., Maazoun,R. & Killick-Kendrick, R. (1984).Ecologicdesleishmanioses dam le sud de la France.18Identification enzymatique de L&.&mania infantum Nicolle, 1908, isolC de Phleboto-
j&;ived
TRANSACTIONSOF THE ROYAL SOCIETYOF TROPICALMEDICINE
HYGIENE(1990) 84, 786
mus ariasi Tonnoir,
1921, spontanement infest6 en
1 Short Report 1 The importance of rapid diagnosis of new cases of cutaneous leishmaniasis in pin-pointing the sandfly vector L. Ryad*, A. Vexenag, P. D. Marsden’, R. Lains~d +d J. J. Shawl ‘The Wellcome Parasitolp p Umt, Carxa Postal 3, t$5*001, Belhn, Par& Bpztl; Ntileo de Medicina Troprcal e Nutripio, Umverstdade de Bra&a, Brash, DF, Brazil
We have searched for many years for the vector of Leiskmania (Viunnia) braziliensis in the endemic area
of T&s Bra$os, Bahia, Brazil. Since over 95.1% of the phlebotomine santies captured in and around houses are Lutzomyia wkitmani (Antunes & Coutinho), and leishmanial ulcers were occasionally seenin &all, house-bound children, this specieswas susnected to be the vector WEXENAT et al., 1986). $&al efforts were made t&dissect sandf3iescaught around the houses of families where more than one caseof acute cutaneous leishmaniasis was present and, although several such attempts failed; we briefli record here the lirst successful isolation of L. (V.) braziliensis from Lu. wkitmuni as a result of this policy. In January, 1986 two men attended the field clinic, both with ulceration of the lower leg of 15 d duration. They were accustomed to meet in the evening, frequently clad in shorts and thong sandles. They discussed the day’s events while seated on a log in front of their adjoining houses in Fazenda Rizada, a
AND
Transactions of the Royal Society of Tropical Medicine and Hygiene, 78, 48U84. 21 May 1990; accepted
12July
farm currently under longitudinal study and where 6 casesof cutaneous leishmaniasis have been recorded during the decade 1980-1990. Parasites later identiiied as L. (V.) braziliensis were isolated from both patients (LTB:865 and LTB:866) and treatment with Glucantime commenced. Two days later entomological studies were initiated and concentrated on this locality: in the next few days 193Lu. wkimni were captured from a chicken-house near the house of one of the patients and, on dissection, one was found to be inf&ted with L..(V.) braziltis (RYAN et al.. 1987). Identification was made in bdth the Be& and ‘Brasilia laboratories, using monoclonal antibodies and enzyme profiles (strain IWHIIBR/86/M10187). The very recent nature of the human infections undoubtedly helped in our selection of an appropriate sandfly capture-site. Usually patients arrive for consultation later in the disease,but on this occasion the farm was near the medical post and the men had a car at their disposal. Later in the sameyear (HOCK et al., 1986) 2 further isolates of L. (V.) braziliensis were made from Lu. whitmani, on a farm where several children were infected in the same family. References
Hoch, A., Ryan, L., Vexenat, J. A., Rosa, A. C. 0. C. & Barreto. A. C. (1986). Isolation of Leishmania brazilknsis braziliekis
and‘o&r trypanosomatidsfrom phleboto-
mines in a mucocutaneousleishmaniasis endemic area in Bahia, Brazil. Mem6rias do Inszinrto Oswald0 Cruz, 81, supplkment?abstract no. Bl 44. Ryan, L., Lamson, R., Shaw, J. J., Braga, R. R. 81 Ishikawa, E. A. Y. (1987). Leishmaniasis in Brazil. XXV. Sandfly vectors of Leishmania in Par6 State, Brazil. Medical and Veterinary Enmwwlogy, 1, 383-395. Vexenat, J. A., Barreto, A. C., Cuba, C. C. & Marsden,P. D. (1986). Caracteristicas epidemioldgicas da leishmaniose tegumentar americana em uma regiao end&mica do Estado da Bahia. III. Fauna flebotomlnica. MenGrius do Institute Oswald0 Crw,
*Present address: Philip Morris USA ResearchCentre, P.O. Box 26583, Richmond, VA 23261, USA. Off-print requests to R. Lainson.
for publicatkm
Received
1990
l2Jun.t~
81, 293-301.
1990; acceptedfmpublication
ZJune