J Vector Borne Dis 49, March 2012, pp. 23–26

Laboratory colonization of Lucilia sericata Meigen (Diptera: Caliphoridae)
strain from Hashtgerd, IranF. Firoozfar1,2, S.H. Mosa Kazemi1, K. Shemshad3, M. Baniardalani1, M. Abolhasani1,
A. Biglarian4, A. Enayati5 & J. Rafinejad1,6
1Department of Medical Entomology and Vector Control, School of Public Health; 2Students Scientific Research Center, Tehran Universityof Medical Sciences, Tehran, Iran; 3Department of Entomology, Science and Research Branch, Islamic Azad University, Tehran; 4Departmentof Biostatistics, University of Social Welfare and Rehabilitation (USWR), Tehran; 5Department of Medical Entomology, School of Public
Health and Health Sciences Research Center, Mazandaran University of Medical Sciences, Sari; 6Evaluation Management and Development
Center, Deputy of Research Ministry of Health and Medical Education, Tehran, Iran

ABSTRACT
Background & objectives: The treatment of wounds with live green bottle fly larvae is receiving considerable
attention in many countries. Laboratory rearing of Lucilia sericata is crucially important for the treatment ofwounds.
Study design: The study was carried out for mass rearing of green bottle flies from April to November 2010.
Hand catch and net trap baited with beef and cattle liver were used to collect adult flies from the field. The
collected samples were placed in appropriately labeled tubes and sent to the laboratory. Adult stage flies rearedin the insectary were used for species identification using specific keys.
Results: A total of 89 flies (55 females and 34 males) were collected from Hashtgerd area. In the first generation,
299 flies were produced in the laboratory including 105 (35.12%) males, and 194 (64.88%) females. The female/
male sex ratio was 1.61 for parents, whereas it was 1.84, 1.30 for F1 and F2 generations respectively. In total,
432 flies were reared in F3 generation including 173 (40.04%) males, and 259 (59.96%) females, and the sex
ratio was 1.49.
Conclusion: Setting up the mass rearing of sheep blowfly at the School of Public Health, Tehran University of
Medical Sciences is an important step in producing candidate flies for the treatment of myiasis by maggot
therapy in future.
Key words Insectary; Iran; Lucilia sericata; maggot therapy

INTRODUCTION
The sheep blowfly Lucilia sericata has a long history
in medicine and its maggots have been used in wound healing. Some unsuccessful surgical and antibiotic treatments
of infections such as temporal mastoiditis and perineal
gangrene were treated using maggot therapy. Wounds resulting from cuts, wounds or even trauma resulting from
diseases like diabetes, bed sores, gangrene, and burns are
the problems of human societies1. In many of these cases,
surgery is the only treatment, sometimes leading to amputation. Leg amputation due to diabetes is frequently reported from around the world. According to World HealthOrganization, this happens every 30 seconds. Diabetic footulcer is responsible in half of diabetic patients’ visits tothe hospitals. In many of these patients, it may lead to
amputation of one or both feet2. Treatment by flies’ larvae as a suitable alternative method in the treatment of thewounds has introduced3. Up to now, maggot therapy was
performed for treating many different kinds of wounds.

Centrally important to maggot therapy is that the larvae used in the treatment of wounds should reduce theinfections and improve the nutrition of tissues for rapid
improvement of wounds. There are 1100 known speciesof sheep flies, with 228 species in the Neotropics, Africa
and southern Europe. Lucilia species has been reported in
the countries, namely India, Japan, central America, andsouthern United States of America4 .
Considering the significance of maggot therapy, establishing an insectary for mass rearing of larvae to provide hospitals and medical centers with Lucilia larvae isimportant. The aim of this study was mass rearing andmaintenance of L. sericata (Phaenicia sericata) maggot
under laboratory conditions at the School of Public Health,Tehran University of Medical Sciences.
MATERIAL & METHODS
Study area
The experimental study was performed from April to24

J Vector Borne Dis 49, March 2012

November 2010. Sampling methods used were hand catchby net and bait traps. Sampling was carried out in places
including gardens, around livestock, slaughterhouse in
Hashtgerd county, Alborz province, northern Iran.
Adult fly collection
The samples were caught from different places ofAlborz province and were sent to the insectary of Medical
Entomology at School of Public Health, Tehran University of Medical Sciences in appropriately labeled tubes5.
Beef and Hamburger were used as bait for collecting adultstage of the fly in open area. Information such as placeand date of collection, sampling method and weather condition was recorded5.
Species identification
Adult flies were identified using standard keys6–8. The
diagnosis of larvae is performed based on the respiratory
pores and posterior of cephalo-pharyngeal skeleton of larvae, therefore, definitive diagnosis was carried out on deadlarvae. In order to identify the live specimens, adults wereanesthetized by CO2, ether or cold shock for a short time.
After identification, the adults were transferred to newcages for oviposition.
Maintenance
The flies were maintained in the insectary under controlled conditions of mean temperature of 27±1°C, relative humidity of 80±5%, and daily light /dark period of
16:8 h. The average temperature for larval development
was also 27±1°C. The electrical fly killer was hung in theinsectary area to kill the free flying flies in the insectary9,10.
Layers of red meat with tissue papers in between wereplaced in the petri dish and, then a cup container with cutedge was placed on the meat11. In order to rear the larvae,
sheep blood agar was used because of its simplicity foodsource12,13. Supervision of rearing cages was essential andlarvae were isolated from rearing cages. Upon emerging,
the adults were placed in new cages and provided withessential food.

to the Tehran School of Public Health. The life cycle ofgreen flies is shown (Figs. 1–4) and the duration of life
cycle of male and female L. sericata is given (Table 1).
The average number of female parents emerged per eggs
batch was 6.1. The time required for the eggs to hatch
was 52±2 h. The mean time for larval and pupal development in F1, F2, and F3 generations was 96±2, and
176±2.33 h respectively. The time for emerging from pupae to laying eggs was 200±2.66 h. In F1 generation, 507
maggots age I were reared to 299 adults, hence, a mortality rate during the F1 generation of 41.03% was reported.
Mortality rate of Lucilia sericata was 53.15% for F2 generation, whereas in F3 generation, it was 27.28 %. Number of eggs, larvae and pupae stages I, II, III and adults
yielded in F1, F2 and F3 generations are shown (Table 2).
Totally, 299 flies were reared including 105 (35.12%)
males, and 194 (64.88%) females in the first-generation.
They were transferred to spawn cages and nine eggs
batches were obtained. In F1 generation, the average number of female flies emerged per eggs batch was 21.55%.

Fig. 1: The batch of eggs of Lucillia sericata at the insectary, School of Public Health, Tehran University of
Medical Sciences.

Statistical analysis
The hatching dishes were filled with larval medium
and were visited daily according to method ofFleischmann13. All values are expressed as the mean ±standard error of the means (SEM).
RESULTSIn all, 89 adult flies were collected from slaughterhouses of Hashtgerd, Alborz province and were transferredFig. 2: The Lucillia sericata maggots at the insectary,
School of Public Health.

25

Firoozfar et al: L. sericata laboratory from Hashtgerd, IranDISCUSSION
The different time periods required for the completionof three generations of Lucilia sericata in our study showedthat the life stags are affected by temperature and humidity of the Insectary. The first generation of wild flies took
longer than their parents to lay eggs. This delay may bejustified by the time required for the adaptation to the captivity conditions. Also the first generation took longer tomate. Lysek in 1991 reported the optimal temperature of
37± 3°C for rearing this fly. The duration of larval stage
was reported to be 4 to 13 days14. In our study, the duration of larval stage was 96 ± 2 h with an average temperature of 27 ± 1°C. Sherman and Wyle11; and Wolff and
Hansson1 colonized this species in insectary and reported
a cycle from egg to adult of about 1–2 weeks a period
which was calculated 8.33 days in our study. In parallel,
the egg to egg period was reported as 2–3 weeks15. In ourstudy, the period from egg to adult was noticeably longer
in F2 generation, which might be justified by different diet,nutrition and new environment. Comparison of the resultsof studies by Sherman and Wyle 11; and Wolff and
Hansson1 indicated that in the second generation in ourstudy less eggs were developed to larvae and adults successfully (46.85%). This decrease in the second generation was more than that in the first generation, this fact
might be explained by the specific conditions including
light, temperature, humidity, food and larvae nutrition. The
effect of different kinds of food on larval development ofFig. 3: The pupa of Lucilia sericata at the insectary,
School of Public Health.

Fig. 4: The adult of Lucilia sericata at the insectary,
School of Public Health.

Table 1. Duration of life stage of Lucilia sericata in the insectary 2010
Generation

Parents
F1
F2
F3

Adult

Egg period

MaleFemaleTime (h)

34
105
84
173

55
194
110
259

–
60 ± 348 ± 148 ± 2

Larval stage

Pupa stage

Mean ± SEM Time (h) Mean ± SEM
–
52±2

–
72 ± 196 ± 3120 ± 2–96 ± 2

Emergence to egg laying (h)Time (h) Mean ± SEM

Time(h)Mean ± SEM–
192 ± 3168 ± 2168 ± 2–312 ± 3168 ± 3120 ± 2–200 ± 2.66–
176 ± 2.33

Table 2. Number of eggs, batch, and larvae stages I, II, III, pupae and adults of Lucilia sericata during
F1, F2 and, F3 generations in insectary 2010
Generation
rationF1F2F3

Egg batchbatch (No.)
9114

Larval stage

Pupae

I

II

III

507 (100)
414 (100)
594 (100)

431 (85)348 (84.05)
553 (93.09)

375 (87)
293 (84.19)
509 (92.04)

Figures in parentheses indicate percentages.

AdultsMale

330 (87)
214 (73.03)
464 (91.15)

105 (35.1)
84 (43.3)
173 (40.1)

Total
Female194 (64.9)
110 (56.7)
259 (59.9)

299
194
432

26

J Vector Borne Dis 49, March 2012

Lucilia sericata is considerable. The size of larvae fed on
hamburger was smaller than those fed on beef and liver.
Similar to our results, the growth rate of Lucilia larvae
fed on beef liver16 was significantly lower than those grownon the chicken flesh and beef. Protein intake is necessary
in the first generation of in vitro rearing. The average period between emerging of adults, mating, spawning, andlaying eggs in our study was 8.33 days, whereas Sherman
and Wyle11; Wallman and Day17 stated two weeks fromemergence to laying of eggs. Mortality rate during the life
cycle of Lucilia sericata was 41.03, 53.15 and 27.28% in
F1, F2, and F3 generations respectively. The mortality
rate of gravid female was more than non-gravid and malein F2 generation. The susceptibility of the flies to temperature and humidity was noted in our study. The stages
I and II larvae are quite sensitive to dryness and food shortage, whereas the III instar larvae are sensitive to humidity. Establishing a colony of this fly in insectary in Iran isof high importance for undertaking maggot therapy to treat
the disease.
ACKNOWLEDGEMENTS
The authors are grateful to Dr Ladonni and Mr.
Akbarzadeh, Department of Medical Entomology and
Vector Control, for their valuable suggestions. The authors are also grateful to Mr Hossaini for his kind cooperation throughout the study in flies’ insectary. This
project was financially supported by a grant from Tehran
University of Medical Sciences Project No. 89-03-2711453. The authors declare that they have no conflictsof interest.

3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.

14.
15.

16.

REFERENCES
1.
2.

Wolff H, Hansson J. Rearing larvae of Lucilia sericata forchronic ulcer treatment – an improved method. Sweden: Department of Dermatology. Acta Derm Venereol 2005; 85: 126–31.
Cetinkaya M, Özkan H, Köksal N, COkun SZ. Neonatal myia-

17.

sis: a case report. Turkish J Pediatrics 2008; 50: 581–4.
Bani-Ardalani M. Weekly Medical Report 2005; No. 578, 15
years.
UniProt.“Calliphoridae”(HTML). Available from: http://
beta.uniprot.org/taxonomy/7371 (accessed on May 31, 2008).
Hall MJ. Trapping the flies that cause myiasis: their responses
to host stimuli. Ann Trop Med Parasitol 1995; 89(4): 333–57.
James MT. The flies that cause myiasis in man. United States
Department of Agriculture Miscellaneous Publication 1974; 631:
1–175.
Zumpt F. Calliphorinae. In: Lindner E, editor. Die fliegen derpalaearktischen Region XI, 64i. Schweizerbart, Stuttgart 1959;
8: 1–140.
House-flies, blow-flies and their allies (Calyptrate: Diptera).
In: Lane RP, Crosskey RW, editors. Medical Insects Arachnids. Chapman and Hall 1993; p. 403–28.
Greenberg B. Flies through history. Flies and Disease, I edn.,
Princeton, New York: Princeton University Press 1973;
p. 1–447.
Sherman RA, My-Tien Tran JM. A simple sterile food source
for rearing the larvae of Lucilia sericata (Diptera: Caliphoridae).
Med Vet Entomol 1995; 9(4): 393–8.
Sherman R, Wyle F. Low-cost, low-maintenance rearing of maggot in hospitals, clinics and schools. Am J Trop Med Hyg 1996;
54: 38–41.
Spiller D. Insect colonization and mass production. In: SmithCN, editor. Houseflies: Chapter 14. New York and London: Academic Press 1996; 203–25.
Fleischmann W, Grassberger M, Sherman R. Maggot therapy: ahandbook of maggot-assisted wound healing complementary
medicine. Stuttgart 93 pp. George Thieme Verlag 14, 70469,
Germany 2004; p. 93. Available from: www.thieme.de
Lysyk TJ. Effects of temperature, food, and sucrose feeding on
longevity of the housefly (Diptera: Muscidae). Environ Entomol
1991; 20: 1176–80.
Nsary J, Ladani M, Mirab-Zadeh A, Mashayekhi M, Sharifi
SH. The final report of the project “Using of larvae Lucilia
Sericata to treatment of infectious wounds. Scientific and Industrial Research Organization of Iran 2006; p. 1–51.
Salimi M, Goodarzi D, Karimfar MH, Edalat H. Human urogenital myiasis caused by Lucilia sericata (Diptera,Caliphoridae) and Wohlfahrtia magnifica (Diptera:
Sarcophagidae) in Markazi Province of Iran. Iranian J Arthropod Borne Dis 2010; 4(1): 72–6.
Wallman JF, Day DM. Influence of food substrate tissue typeon larval growth in Calliphora augur and Lucilia cuprina(Diptera: Caliphoridae). J Forensic Sci 2006; 51: 57–63.

Correspondence to: Dr Javad Rafinejad, Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences and Evaluation Management and Development Center, Deputy of Research Ministry of Healthand Medical Education, P.O.Box 14155-6446, Tehran, Iran.
Received: 2 September 2011

Accepted in revised form: 18 February 2012