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Laboratory colonization of Phlebotomus perfiliewi (Parrot)
876
TRANSACTONS OF THE ROYAL SOCIETYOF TROPICALMEDICINE AND HYGIENE, VOL. 77, No. 6 (1983). CORRESPONDENCE
Asia and Pacific (WHO mimeographed document WHO/ VBC/IO. 786).
Leishmunia major Yakimoff et Schokhor, 1914 (Kinetoplastida, Trypanosomatidae) dam le Sud marocain. ~logagty Rendus de I’Acadknie des Sciences (Paris), 294,
Accepted for publication 5th March, 1983. Accepted for publication 6th April,
Isolation of Leiskmania major Yakimoff & Schokhor, 1914 from Psammomys obesus Gretzschmar, 1828 (Rodentia: Gerbillidae) in Algeria
An important outbreak of cutaneous leishmaniasis has recently been reported (BELAZZOUG,1982) in M’sila. situated in the steppe reaion of the Aleerian northern Sahara.2,103 caskshavebeen recorded. The search for the animal reservoir resulted in the discovery of three rodents? Psammomys obesus Gretzs&mar, 1828, with lesions of the ear margin; impression smears showed Leiskmania amastigotes (BELAZZOUG,1983). Two stocks isolated from a rodent and from a patient having cutaneous leishmaniasis have been typed isoenzymatically at the Laboratoire d’Ecologie Medicale (Prof. J. A. Rioux) of Montpellier (France). Eight enzymes were used (PGM, GPI, G-6-PDH, 6-PGDH, IDH, MDH, ME and GOT) on starch-gel electrophoresis -and showed that both stocks were identical to Leiskmania major from the USSR. P. obesus, already found naturally infected with Leiskmania m Libya (ASHFORDet al., 1977), may be a reservoir of L. mjor in North Africa together with Meriones skawi (DUVERNOY,1842) found infected in South Morocco (RIOIJX et al., 1982). Indirect findings also strongly implicate these rodents in the epidemiology of cutaneous leishmaniasis: in the area where the epidemic occurred, there are numerous burrows of Psammomys, under the Chenopodiaceae, and Meriones, under the bushes of Zizypkus, and even very near the farms built by agricultural developers. Moreover, we captured Pklebotomus papatasi (Scopoli, 1786), the sand-fly vector of cutaneous leishmaniasis in Algeria, on oiled papers placed at the entrance of the burrows. S. BELAZZOUG d’Rpi&miologie, Institut Pasteur, Rue Dr. Laveran, Algiers, Algeria
Service
References Ashford, R. W., Schnur, L. F., Chance, M. L., Samaan, S. A. & Ahmed, H. N. (1977). Cutaneous leishmaniasis in the Libyan Arab Republic: preliminary ecological findings. Annals of Tropical Medicine and Parasitology,
71, 265-271.
Belazzoug, S. (1982). Une epidemic de leishmaniose cutanee dans la region de M’sila (Algerie). Bulletin de la
Socikti de Patholoaie Exotique, 75, 497-504.
Belazzouq, S. (1983). Le nouveau foyer de leishmaniose cutanee de M’sila (Algerie). Infestation naturelle de Psammomys obesus (Rongeur, Gerbillide). Bulletih de la Socikti de Patholo ‘e Exotique, 76, 146-149. ., Akalay, O., Lanotte, G., Ouazzani, Rioux, J. A., Petter, Fg” A., Seguignes, M. & Mohcine, A. (1982). Merionesshawi (Duvernoy, 1842) (Rodentia, Gerbillidae), reservoir de
Laboratory
colonizati;iii)
1983.
Pklebotomus perfliewi
Previous studies by .CO~~DETTI (1962) strongly indicate that Pklebotomus msrtiliewi is the vector of cutaneous leishmaniasis in It&y, but this view, based on epidemiological data, has not yet been demonstrated experimentally. In an investigation of phlebotomine sandflies in Grosset0 province (Tuscany) by MAROLI & BETTINI (1977), P. perfiliewi was also suspected to be a vector of visceral leishmaniasis in some Darts of the area . studied. Recently GRADONI et al. (1983, in press) have demonstrated that in nature P. perfliewi is attracted to Rattus rattus and that this sandflv snecies became infected when fed on R. rattus experimentally infected with Leiskmania infantum. WARD et al. (1981). usine starch eel electrouhoresis and comparing- the &nple~ of I’. &rfiliewi f;om the Tyrrenian side of the Apennine mountains with those from the Adriatic side. showed sitificant variation between enzyme genotypes of pho
colony establishment and mainet tenance are those described bv KILLICK-KENDRICK al. (1977a) and by Maroli et al. (1983, in preparation). Larvae and adults are reared at 28 to 29°C; photoperiod LD 17:7. Larval food is a mixture of 50% rabbit faeces. 25% dried Dabknia and 25% sand (READY & CLOSET, 1980). Adult females readily take blood from hamsters and in a preliminary trial on membrane feeding 38 females (40%) took blood from membrane (chick& skin and‘ rabbit blood). Mating occurs more frequently between males and freshly engorged females than before engorgement. Thus, as reported by GEMETCHU (1976), to obtain a good insemination rate both sexesare left together for 24 hours after a blood meal.
TRANSACTONSOF THE ROYAL SOCErv OF TROPICALMEDICINE AP~‘DHYGIENE, VOL. 77, No. 6 (1983). CORRESFONDENCE 877
The mean time of development from blood meal to emergenceof first adult/pot of the next generation is 47 days (43 to 51). Mean times of development are: gestation, 7 days (6 to 9); hatching of eggs, 6 days (5 to 8); larval development, 22 days (19 to 25); pupal development, 7 days (6 to 9). Under these rearing conditions no diapause was observed although a few larvae showed delay in their development. In fact 14.5% of the fourth-instar larvae in the first generation and 3.5% in the second generation reached the pupal stage after about 60 days. Productivity of the colony was, for the first generation, 885 adults (455 males and 408 females) and for the second generation, 752 adults (402 males and 350 females), from 113 gravid females. In previous attempts, P. perjiliewi could not be bred beyond the first generation (KILLICKKENDRICK et al., 1977b). It seems from our data that
this species,now in the six generation, can be bred in a laboratory colony beyond subsequent generations. P. perj2iewi can thus also be numbered among the Old World speciesof Phlebotomus bred in numbers in the laboratory which, as reported by KILLICKKENDRICK (1979) are argentipes, pemiciosus, longipes
and papatasi. Thanks are due to Dr. E. Pozio who helped in collecting the flies and Dr. L. Gradoni who has given skilled assistancein initial work in the laboratory. The expert technical assistanceof Mr. S. Fiorentino and Mr. E. Guandalini in maintainine the colonv is appreciated and acknowledged. y _I
M. MAROLI Dept. of Parasitology, Secti$tr$ Vector Biology
Is&to
St&iore
and
di San&
Viale Regina Elena 00161 Rome, Italy
no 299, References
Corradetti, A. (1962). Phlebotomus and leishmaniasis in North-Central Italy (Apennine Region). Scientific Report Istituto Superiore di Saniti, 2, 103-109. Gemetchu, T. (1976). The biology of a laboratory colony of Phlebotomus lot&es Parrot & Martin (Diptera: Phlebotomidae). 3oumal of Medical Entomology, 12, 661-671. Gradoni,, L. Pozio, E., Gramiccia, M., Maroli, M. N. & Bettmi, S. (1983). Leishmaniasis in Tuscany (Italy): VII, Studies on the role of the black rat, Rattus rutrus, in the epidemiology of visceral leishmaniasis. Transactions of the T2y-$3%ciety
of Tropical Medicine and Hygiene,
77,
Kiliick-kenhrick, R., Leaney, A. J. & Ready, P. D. (1977a). The establishment, maintenance and productivity of a laboratory, colony of Lutzomyiu longipulpis (Diptera: Psychodidae). Journal of Medical Entomology, 13, 429-440. Killick-Kendrick, R., Ready, I’. D. & Pampiglione, S. (1977b). Notes on the prevalence and host preferencesbf Phlebotomus perfdiewi in Emilia Romagna, Italy. Ecologic des Leishmuniases. Colloques Internutionuux
du CNRS,
239, 169-175. Killick-Kendrick, R. (1979). Biology of Leishmuniu in phlebotomine sandflies, in: Biology of Kinetoplustidu, Vol. 2. Lumsden, W. H. R. and Evans, D. A. (Editors). London: Academic Press, pp. 396-449. Maroli, M. N. & Bettini, S. (1977). Leishmaniasis in Tuscany (Italy): (I) An investigation of phlebotomine sandflies in Grosset0 Province. Trunsucrions of the Rq~ul Society of Tropical Medicine and Hygiene, 71, 315-321.
Ready, P. D. & Croset, H. (1980). Diapause and laboratory breeding of Phlebototnus bemiciosusNewstead and Phlebotomus- ariusi Tonnoir a(Diptera: Psvchodidae) from southern France. Bulletin of Entomological Research, 70, 511-523. Ward, R. D., Bettini, S., Maroli, M., McGarry, J. W. & Draper, A. (1981). Phosphog!ucomutasepolymorphism in Phlebotomus petfiliewi perfiliewi Parrot (Diptera:
Psychodidae)from Centraland northern Italy. AnnaIs of 75, 653-661.
Tropical Medicine and Purasitology,
Accepted for publication
Duffy
14th May,
1983.
blood group distribution
in Malawi
I have recently carried out a survey of the Duffy blood group distribution in Malawi, a densely populated area of southern Africa not previously studied. 158 specimens from Malawians attending the Queen Elizabeth Central Hospital, Blantyre, were tested with anti Fya and anti Fyb sera by a standard albumin spin Coombs’ technique (reagents produced by the Northern Regional Blood Transfusion Service, Newcastle upon Tyne, UK) with the following results: 157 ($.$bj
Fyl;;I W
,9-b+
1 (0:6;)
There is a very high incidence of FfVb- -98.8% or gene frequency 0.994, which is similar to findings in some parts of neighbouring countries of Mozambique, Zambia and Zimbabwe (MOURANTet al., 1976). This was not a study of a single community but most people would be of Chewa origin. The Duffy Fy”+ or FYbc blood group antigen is associated with attachment of Plasmodium tinax merozoites to the red cell (MILLER et al., 1976) and the findings explain the absenceof benign tertian malaria in this area. During 1978-80about 40,000 fihns were examined for malaria at this hospital with over 1,000 positives which were al18yrted as casesof P. falciparum malaria (PAUL, B. PAUL National Blood Transfusion Service, Regional Transfusion Cetttre, Westgate Road, Newcastle upon Tyne NE4 6QB, England. References
Miller, L. H., Mason, S. J., Clyde, D. F. (1976). The resistance factor to Plavnodium &ax in Blacks. The
Duffy-blood-groupgenotype,FylFy. New EnglandJour-
MI of Medicine, 295, 302-304.
Mourant, A. E., Kope~,A. C. & Domaniewska-Sobczak. (1976). The Distributwn of Humun Blood Groups and other Polymorphisms. London: Oxford University Press, p. 593.
Paul,B. (1981).Rain andanaemia-the seasonalvariationof malaria and megaloblastosis.Journul of the Medical Association of Malawi,
8, 19-20.
Accepted for publication
23rd May,
1983.
TRANSACTONS OF THE ROYAL SOCIETYOF TROPICALMEDICINE AND HYGIENE, VOL. 77, No. 6 (1983). CORRESPONDENCE
Asia and Pacific (WHO mimeographed document WHO/ VBC/IO. 786).
Leishmunia major Yakimoff et Schokhor, 1914 (Kinetoplastida, Trypanosomatidae) dam le Sud marocain. ~logagty Rendus de I’Acadknie des Sciences (Paris), 294,
Accepted for publication 5th March, 1983. Accepted for publication 6th April,
Isolation of Leiskmania major Yakimoff & Schokhor, 1914 from Psammomys obesus Gretzschmar, 1828 (Rodentia: Gerbillidae) in Algeria
An important outbreak of cutaneous leishmaniasis has recently been reported (BELAZZOUG,1982) in M’sila. situated in the steppe reaion of the Aleerian northern Sahara.2,103 caskshavebeen recorded. The search for the animal reservoir resulted in the discovery of three rodents? Psammomys obesus Gretzs&mar, 1828, with lesions of the ear margin; impression smears showed Leiskmania amastigotes (BELAZZOUG,1983). Two stocks isolated from a rodent and from a patient having cutaneous leishmaniasis have been typed isoenzymatically at the Laboratoire d’Ecologie Medicale (Prof. J. A. Rioux) of Montpellier (France). Eight enzymes were used (PGM, GPI, G-6-PDH, 6-PGDH, IDH, MDH, ME and GOT) on starch-gel electrophoresis -and showed that both stocks were identical to Leiskmania major from the USSR. P. obesus, already found naturally infected with Leiskmania m Libya (ASHFORDet al., 1977), may be a reservoir of L. mjor in North Africa together with Meriones skawi (DUVERNOY,1842) found infected in South Morocco (RIOIJX et al., 1982). Indirect findings also strongly implicate these rodents in the epidemiology of cutaneous leishmaniasis: in the area where the epidemic occurred, there are numerous burrows of Psammomys, under the Chenopodiaceae, and Meriones, under the bushes of Zizypkus, and even very near the farms built by agricultural developers. Moreover, we captured Pklebotomus papatasi (Scopoli, 1786), the sand-fly vector of cutaneous leishmaniasis in Algeria, on oiled papers placed at the entrance of the burrows. S. BELAZZOUG d’Rpi&miologie, Institut Pasteur, Rue Dr. Laveran, Algiers, Algeria
Service
References Ashford, R. W., Schnur, L. F., Chance, M. L., Samaan, S. A. & Ahmed, H. N. (1977). Cutaneous leishmaniasis in the Libyan Arab Republic: preliminary ecological findings. Annals of Tropical Medicine and Parasitology,
71, 265-271.
Belazzoug, S. (1982). Une epidemic de leishmaniose cutanee dans la region de M’sila (Algerie). Bulletin de la
Socikti de Patholoaie Exotique, 75, 497-504.
Belazzouq, S. (1983). Le nouveau foyer de leishmaniose cutanee de M’sila (Algerie). Infestation naturelle de Psammomys obesus (Rongeur, Gerbillide). Bulletih de la Socikti de Patholo ‘e Exotique, 76, 146-149. ., Akalay, O., Lanotte, G., Ouazzani, Rioux, J. A., Petter, Fg” A., Seguignes, M. & Mohcine, A. (1982). Merionesshawi (Duvernoy, 1842) (Rodentia, Gerbillidae), reservoir de
Laboratory
colonizati;iii)
1983.
Pklebotomus perfliewi
Previous studies by .CO~~DETTI (1962) strongly indicate that Pklebotomus msrtiliewi is the vector of cutaneous leishmaniasis in It&y, but this view, based on epidemiological data, has not yet been demonstrated experimentally. In an investigation of phlebotomine sandflies in Grosset0 province (Tuscany) by MAROLI & BETTINI (1977), P. perfiliewi was also suspected to be a vector of visceral leishmaniasis in some Darts of the area . studied. Recently GRADONI et al. (1983, in press) have demonstrated that in nature P. perfliewi is attracted to Rattus rattus and that this sandflv snecies became infected when fed on R. rattus experimentally infected with Leiskmania infantum. WARD et al. (1981). usine starch eel electrouhoresis and comparing- the &nple~ of I’. &rfiliewi f;om the Tyrrenian side of the Apennine mountains with those from the Adriatic side. showed sitificant variation between enzyme genotypes of pho
colony establishment and mainet tenance are those described bv KILLICK-KENDRICK al. (1977a) and by Maroli et al. (1983, in preparation). Larvae and adults are reared at 28 to 29°C; photoperiod LD 17:7. Larval food is a mixture of 50% rabbit faeces. 25% dried Dabknia and 25% sand (READY & CLOSET, 1980). Adult females readily take blood from hamsters and in a preliminary trial on membrane feeding 38 females (40%) took blood from membrane (chick& skin and‘ rabbit blood). Mating occurs more frequently between males and freshly engorged females than before engorgement. Thus, as reported by GEMETCHU (1976), to obtain a good insemination rate both sexesare left together for 24 hours after a blood meal.
TRANSACTONSOF THE ROYAL SOCErv OF TROPICALMEDICINE AP~‘DHYGIENE, VOL. 77, No. 6 (1983). CORRESFONDENCE 877
The mean time of development from blood meal to emergenceof first adult/pot of the next generation is 47 days (43 to 51). Mean times of development are: gestation, 7 days (6 to 9); hatching of eggs, 6 days (5 to 8); larval development, 22 days (19 to 25); pupal development, 7 days (6 to 9). Under these rearing conditions no diapause was observed although a few larvae showed delay in their development. In fact 14.5% of the fourth-instar larvae in the first generation and 3.5% in the second generation reached the pupal stage after about 60 days. Productivity of the colony was, for the first generation, 885 adults (455 males and 408 females) and for the second generation, 752 adults (402 males and 350 females), from 113 gravid females. In previous attempts, P. perjiliewi could not be bred beyond the first generation (KILLICKKENDRICK et al., 1977b). It seems from our data that
this species,now in the six generation, can be bred in a laboratory colony beyond subsequent generations. P. perj2iewi can thus also be numbered among the Old World speciesof Phlebotomus bred in numbers in the laboratory which, as reported by KILLICKKENDRICK (1979) are argentipes, pemiciosus, longipes
and papatasi. Thanks are due to Dr. E. Pozio who helped in collecting the flies and Dr. L. Gradoni who has given skilled assistancein initial work in the laboratory. The expert technical assistanceof Mr. S. Fiorentino and Mr. E. Guandalini in maintainine the colonv is appreciated and acknowledged. y _I
M. MAROLI Dept. of Parasitology, Secti$tr$ Vector Biology
Is&to
St&iore
and
di San&
Viale Regina Elena 00161 Rome, Italy
no 299, References
Corradetti, A. (1962). Phlebotomus and leishmaniasis in North-Central Italy (Apennine Region). Scientific Report Istituto Superiore di Saniti, 2, 103-109. Gemetchu, T. (1976). The biology of a laboratory colony of Phlebotomus lot&es Parrot & Martin (Diptera: Phlebotomidae). 3oumal of Medical Entomology, 12, 661-671. Gradoni,, L. Pozio, E., Gramiccia, M., Maroli, M. N. & Bettmi, S. (1983). Leishmaniasis in Tuscany (Italy): VII, Studies on the role of the black rat, Rattus rutrus, in the epidemiology of visceral leishmaniasis. Transactions of the T2y-$3%ciety
of Tropical Medicine and Hygiene,
77,
Kiliick-kenhrick, R., Leaney, A. J. & Ready, P. D. (1977a). The establishment, maintenance and productivity of a laboratory, colony of Lutzomyiu longipulpis (Diptera: Psychodidae). Journal of Medical Entomology, 13, 429-440. Killick-Kendrick, R., Ready, I’. D. & Pampiglione, S. (1977b). Notes on the prevalence and host preferencesbf Phlebotomus perfdiewi in Emilia Romagna, Italy. Ecologic des Leishmuniases. Colloques Internutionuux
du CNRS,
239, 169-175. Killick-Kendrick, R. (1979). Biology of Leishmuniu in phlebotomine sandflies, in: Biology of Kinetoplustidu, Vol. 2. Lumsden, W. H. R. and Evans, D. A. (Editors). London: Academic Press, pp. 396-449. Maroli, M. N. & Bettini, S. (1977). Leishmaniasis in Tuscany (Italy): (I) An investigation of phlebotomine sandflies in Grosset0 Province. Trunsucrions of the Rq~ul Society of Tropical Medicine and Hygiene, 71, 315-321.
Ready, P. D. & Croset, H. (1980). Diapause and laboratory breeding of Phlebototnus bemiciosusNewstead and Phlebotomus- ariusi Tonnoir a(Diptera: Psvchodidae) from southern France. Bulletin of Entomological Research, 70, 511-523. Ward, R. D., Bettini, S., Maroli, M., McGarry, J. W. & Draper, A. (1981). Phosphog!ucomutasepolymorphism in Phlebotomus petfiliewi perfiliewi Parrot (Diptera:
Psychodidae)from Centraland northern Italy. AnnaIs of 75, 653-661.
Tropical Medicine and Purasitology,
Accepted for publication
Duffy
14th May,
1983.
blood group distribution
in Malawi
I have recently carried out a survey of the Duffy blood group distribution in Malawi, a densely populated area of southern Africa not previously studied. 158 specimens from Malawians attending the Queen Elizabeth Central Hospital, Blantyre, were tested with anti Fya and anti Fyb sera by a standard albumin spin Coombs’ technique (reagents produced by the Northern Regional Blood Transfusion Service, Newcastle upon Tyne, UK) with the following results: 157 ($.$bj
Fyl;;I W
,9-b+
1 (0:6;)
There is a very high incidence of FfVb- -98.8% or gene frequency 0.994, which is similar to findings in some parts of neighbouring countries of Mozambique, Zambia and Zimbabwe (MOURANTet al., 1976). This was not a study of a single community but most people would be of Chewa origin. The Duffy Fy”+ or FYbc blood group antigen is associated with attachment of Plasmodium tinax merozoites to the red cell (MILLER et al., 1976) and the findings explain the absenceof benign tertian malaria in this area. During 1978-80about 40,000 fihns were examined for malaria at this hospital with over 1,000 positives which were al18yrted as casesof P. falciparum malaria (PAUL, B. PAUL National Blood Transfusion Service, Regional Transfusion Cetttre, Westgate Road, Newcastle upon Tyne NE4 6QB, England. References
Miller, L. H., Mason, S. J., Clyde, D. F. (1976). The resistance factor to Plavnodium &ax in Blacks. The
Duffy-blood-groupgenotype,FylFy. New EnglandJour-
MI of Medicine, 295, 302-304.
Mourant, A. E., Kope~,A. C. & Domaniewska-Sobczak. (1976). The Distributwn of Humun Blood Groups and other Polymorphisms. London: Oxford University Press, p. 593.
Paul,B. (1981).Rain andanaemia-the seasonalvariationof malaria and megaloblastosis.Journul of the Medical Association of Malawi,
8, 19-20.
Accepted for publication
23rd May,
1983.