Academic Journal of Entomology 5 (2): 08-2, 202 ISSN 995-8994 IDOSI Publications, 202 DOI: 0.5829/idosi.aje.202.5.2.6398 A Study on Pupation of Calliphorid and Sarcophagid Towards Different Applied Substrates 2 2 Abdul Ghafoor, Maryam Siddique, Sohail Khaliq, 2 3 Faiza Mureed and Syed Mohsin Bukhari Department of Wildlife and Fisheries, GCU Faisakabad, Pakistan 2 Department of Chemistry, University of Agriculture, Faisalabad, Pakistan 3 Nuclear Institute for Agriculture and Biology, Faisalabad, Pakistan Abstract: Different substrates were used to evaluate e pupation site preferences of Calliphorid and Sarcophagid flies released in two batches of 0 and 50 during e mons of October and December. Data were recorded for fifteen minutes for consective five days. In e mon of October batches of 0 were found most attracted by e feaers (5.8±6.65 day ) and least by e bread crumb (4.6±6.885 day ) while e batches of 50 showed more response towards soil (5.4±6.897 day ) and least towards e bread crumb (4.8±6.709 day ). During e mon of December, most and least attractive substrates for e batches of 0 and 50 were recorded e cotton (4.0±6.898 day ) and feaers (5.6±5.485 day ), respectively. Pupation substrates showed statistical significant variations (p<0.00) between em. Mons did not differ significantly (p>0.00). By comparing bo batches and mons it was concluded at feaers (5.8±6.65 day ) proved e most attractive substrates while bread crumb (4.4±2.08 day ) least preferred site for pupation. Key words: Pupation Attraction Substrates Blow Flesh Flies INTRODUCTION black, blue, copper and Green []. Blowflies often found from large to moderate in size. Adult blowflies feed Blowflies, flesh flies, beetles, mites, mos and primarily on flower nectar, plant sap and oer surgery bacteria are dominant groups of organisms which play a materials [2]. The members of Sarcophagidae are also very important role in decomposition of materials of called flesh flies. These are similar to blow flies in bo plant and animal origin. A number of non-feeding flies larval and adult habits. Flesh flies generally complete eir are scavengers in nature but e most important among life cycle wiin 8-2 days. Larvae of flesh flies resemble ese are blow flies (Calliphoridae) and flesh flies e larvae of blowflies and go rough ree larval instars. (Sarcophagidae). These flies, especially, play a much The larval skin during e pre-pupal stage contracts and important role in decomposition. Pupae of bo flies are hardens into a protective shell. This shell is called a resistant to environmental extremes. Immature stages of puparium. Adult flies are grayish wi ree black strips flies as post feeding larva or pre-pupa disperse to find running e leng of e top surface of orax []. suitable site to pass pupal stage. The adults of ese flies are known to be of great The members of Calliphoridae are called e blowflies. medical and veterinary importance due to eir role in Their life cycle, generally, completes wiin 9-2 days and myiasis in wild and domestic animals [3]. Blow flies and come in four stages. Larvae or maggots are also white to flesh flies as hosts for different parasitoids are discussed. yellowish when fully grown, ey move around e corpse Maggots are also used in food production. Larvae are as a group and pre-pupae migrate away from corpse introduced to cheese at e time of advanced level of seeking a suitable pupation site. Blowfly s pupae are fermentation and for breaking down e cheese s fats. encased in light brown to black puparia, whereas, adult Maggots can cause allergic reactions, toxicity, stomach, flies are recognized by eir metallic colors ranging form intestinal problems, diarrhea, vomiting, nausea and Corresponding Auor: Dr. Abdul Ghafoor, Department of Wildlife and Fisheries, GCU Faisakabad, Pakistan. 08
Acad. J. Entomol., 5 (2): 08-2, 202 dysentery. Immature stages of blow and flesh flies are one RESULTS of e primary invertebrate consumers of decomposing animal organic matter and human dead body [4]. Larvae or Abundance of pre-pupae of Calliphorid and maggots migrate away from corpse in order to find Sarcophagid flies preferring a particular pupation was suitable site to pass pupal stage. This pupal stage is observed against six different pupation substrates. These extremely important in forensic entomology. So it is substrates were commonly found in e vicinity of human necessary to investigate pattern of larval dispersion on habitats. This experiment was designed to trace e e pupation site. The pupation site preference has been forensically important pre-pupae wi reasonable studied because of number of larvae pupated at different accuracy. Variations among e all six pupation substrates sites in e culture [5]. were differing significant (p<0.00). Significant variations were observed between e batches (p<0.0) but MATERIALS AND METHODS significant differences (p>0.00) were not found between e mons (Table ). Interactions between pupation The experiment trials were conducted in open field of substrates and batches (PSXB) and batches and mons Zoology Department at G.C. University, Faisalabad. (BXM) showed significant variations (p<0.00) between The pre-pupae of Calliphorid and Sarcophagid flies were em and interaction between pupation substrates and secured from e chicken corpse put in a blowfly trap [6]. mons (PSXM) exhibited e non-significant variations Different pupation substrates were placed in e circular (p>0.00) between em. Second order interactions among manners wi few inches distance. The responses of pupation substrates, batches and mons (PSXBXM) pre-pupae of Calliphorid and Sarcophagid flies were revealed e statistical significant differences (p<0.00) observed towards different pupation substrates. The among em (Table ). pre-pupae were released in two batches of 0 and 50 Cotton and soil substrates attracted (26 d, 25 d ) pre-pupae in e center of a four feet diameter hard board. st nd about e same number of pre-pupae in e and 2 day. The experiment was conducted in two mons (October Semi-burnt substrate (54 d ) also attracted e same and December) in order to observe e difference wi e nd number of pre-pupae from 2 to 5 day. Inconsistent reference to environmental factors changing in bo patterns of attractions for five days were observed in e mons and to confirm e data recorded. The choice, cases of cotton bread crumb and cracked bark. Minimum considered to be made when pre-pupae reached a (zero) and maximum (26 d ) pre-pupae recorded for e particular pupation substrate, was recorded. It was also st soil and cotton during e day, respectively. Similarly observed, for fifteen minutes, at wheer e choice persistent increasing patterns were observed for cotton made by pre-pupae was temporary or permanent. The and breadcrumb while reverse was true for soil. Maximum number of pre-pupae making particular choices was (7 5d ) and minimum (23 5d ) pre-pupae were observed recorded. There were six pupation substrates viz. Soil for e cases of feaers and breadcrumb and differed (SOL); Semi-burnt substrate ( SBS); Feaers (FTR); significantly (p<0.00). Statistically significant variations Cotton (CTN); Bread crumb (BRC) and cracked bark (p>0.00) did not found for soil, cotton and bread crumb (CBR). (Table ). Feaers (79 5d ) proved most attractive The observations were made daily from 9:00 am to pupation site and least (23 5d ) for bread crumb :30 am up to five consecutive days. The whole (Table 3). experiment was repeated in October and December in Responses of bo flies towards various substrates order to determine eir responses on dispersal behavior were observed. Progressive increasing or decreasing of pre-pupae. A plastic bucket of 5 inches dep was patterns of attractions did not found for all pupation also used during e experiment in order to avoid e loss materials. Only increase in pre-pupae was seen in e case of pre-pupae or post feeding larvae by putting e trap nd of breadcrumb from e 2 day to 4 day. Significant inside e bucket. Data was analyzed by using ree factor fluctuations were recorded for all pupation substrates factorial (batches, pupation substrates and mons) and during e whole period of study (Table 3). Maximum and completely randomized design [7]. Analysis of variance minimum prepupae were seen in e case of cotton and (ANOVA) was employed for e data recorded and bread crumb, respectively. Significant variations (p<0.00) differences among means were determined [8]. found between em (Table ). Pre-pupae hiding under e 09
Acad. J. Entomol., 5 (2): 08-2, 202 Table : Total number of pre-pupae of five days; mean values per day (mean±sd) and batches of 0 and 50 during e mon of October. BATCHES OF 0 BATCHES OF 50 ---------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------- P.S. D D2 D3 D4 D5 T Mean±SD D D2 D3 D4 D5 T Mean±SD SOL 0 25 5 0 6 2.2±7.477ab 8 2 22 6 0 77 5.4±6.897a SBM 4 0 0 0 0 54 0.8±.552bc 7 6 6 6 9 44 8.8±6.638cd FTR 5 0 6 4 79 5.8±6.65a 20 2 0 8 2 62 2.4±6.74bc CTN 26 0 7 5 59.8±7.56bc 25 3 9 2 4 73 4.6±6.739b BRC 5 3 4 7 23 4.6±6.885d 9 2 6 7 0 24 4.8±6.709e CBR 2 8 3 5 0 68 3.6±6.70a 4 0 6 3 3 56.2±6.508c CV(%) LSD vb b2 CV LSD vb b2 2.40 0.20-0.633 0.888 0.20 8.20-0.633-0.700 Soil (SOL); semi-burnt material (SBM); Feaers (FTR); cotton (CTN); bread crumb (BRC); cracked bark (CBR); D, days; mean values followed by same alphabets do not differ at (p 0.0); T, total pre-pupae of five days; b, Skewness (lack of symmetry in data distribution); b 2, Kurtosis (showing peak and leng of tail in data distribution); P.S. pupation substrate Table 2: Total number of prepupae of five days; mean values per day (mean±sd) and batches of 0 and 50 during e mon of December. BATCHES OF 0 BATCHES OF 50 --------------------------------------------------------------------------------------------------- ---------------------------------------------------------------------------------- P.S. D D2 D3 D4 D5 T Mean±SD D D2 D3 D4 D5 T Mean±SD SOL 2 4 3 4 2 65 3±6.3230b 6 2 4 3 5 70 4±05.63a SBM 8 2 0 9 50 0±6.220c 2 3 2 3 6 2.2±5.509b FTR 4 2 3 6 5 70 4±6.0080a 7 4 6 4 7 78 5.6±5.485a CTN 6 6 4 3 70 4±6.8980a 6 4 5 6 72 4.4±5.525a BRC 4 3 8 7 9 3 6.2±5.8540d 5 6 6 0 5 22 4.4±02.08d CBR 2 2 8 9 0 5 0.2±2.753c 2 2 0 8 2 5 0.2±5.338c CV (%) LSD vb b2 CV LSD ` vb b2 8.77 9.74-0.5223-0.7498 0.404 0.40 0.344.387 Soil (SOL); semi-burnt material (SBM); Feaers (FTR); cotton (CTN); bread crumb (BRC); cracked bark (CBR); mean values followed by same letters do not differ at (p 0.0 ) ; D, days; T, total prepupae of five days;, Skewness (lack of symmetry in data distribution); Kurtosis, b 2 (displaying peak and tail of data distribution); P.S. pupation substrate Table 3: Analysis of variance (ANOVA) of data for pupation site preference of Calliphorid and Sarcophagid pre-pupae towards different substrates in e batches of 0 and 50 during e mons of October and December. SOV df SS MS Batches (B) 3.250 3.25*** Pupation materials (PM) 5 353.45 270.692*** BXPM 5 50.792 0.58*** Mons (M) 2.3470 2.347 NS BXM.250.25 NS PMXM 5 48.903 9.78*** BXPMXM 5 59.792.958*** Error 48 07.333 2.236 Total 7 ***, Significant differences (p<0.00); NS, non-significant differences (p>0.00) soil (5.4±6.897 day ) and cotton (4.6±6.739 day ) did not differ significantly (p>0.00). Similar cases were observed for feaers and cracked bark. Cracked bark attracted same no of pre-pupae for 4 and 5 day and maximum attraction found as 56 5day. In e batches of 50 during October, soil (77 5d ) proved cist preferable en bread crumb (24) (Table 3). Considerable increasing trends were observed for breadcrumb and cracked bark while decreasing for soil and cotton substrates. Fluctuations patterns were seen in all e remaining materials. About same pre-pupae were attracted by e feaers and cotton pupation materials. Maximum (70/5d) attractions were found in e cases of feaers and cotton respectively while minimum (3/5d) for bread-crumb material. Remaining and cotton substrates attracted same number of pre-pupae and found non-significant variations (p>0.00) between em (Table ). Highest attraction (70 5d ) recorded for e feaers and cotton and least (3 5d ) attracted by e bread crumbs material (Table 4). During December in e batches of 50 different pupation substrates exhibited different patterns if distribution. Soil and semi-burnt material contained considerable increasing trends while consistent patterns were observed for e bread crumb material. Maximum number of pre-pupae found for e rd feaers and cotton in e 3 and 4 day of study (Table 2). As a whole maximum (78 5d and minimum (22 5d ) number of pre-pupae maximum recorded for e feaers and breadcrumb in e mon of December and 0
Acad. J. Entomol., 5 (2): 08-2, 202 significant variations were observed between em. In e observed by many investigators [, 5, 2]. Similarly, e contrary, values did not show any significant differences research on pupation site preference of blow and (p> 0.00) for soil, feaers and cotton. During e mon flesh flies were also carried out by numerous if December in e batches of 50 feaers (78 5d ) eminent researchers [5, 4, 6, 7]. In e present found best for pre-pupae and least bread crumb (22 5d ). research on various pupation substrates larvae in batches of 0 and 50 were released and eir dispersal DISCUSSION behaviors was observed during e mons of October and December. Goff, [8] pointed out at blowflies Adult of blow and flesh flies are known to be of e arrived wiin minutes to several hours after dea great medical and veterinary importance due to eir role depending on environmental conditions but flesh flies in e myasis in wild and domestic animals [3]. Myasis in arrived at e same time or several hours after blow humans commonly detected in neck or scalp regions, but flies. During e mon of October feaers (79 5d ) may occur in oer parts of body as well. Serious chronic proved e most preferred substrate for pupation but effects and dea have resulted from infestation of e Yasin [5] found at soil and semi-burnt material was e nose, eyes, ears and mou. Immature stages of bo flies most preferred pupation site. Various researchers proved might be attacked by parasitoids or predators like ants, at at bark was e most attractive site for pupation spiders and crows. Several species of blowflies and flesh [2,6,7]. Sometimes Calliphorid and Sarcophagid flies flies transport a variety of gastrointestinal paogens and like e soil for residing due to its soft nature. Dry leaves are carrier of number of diseases. In e medical research and newspapers provided e moderate attraction for maggots erapy has been used for various purposes like pre-pupae [6, 7]. Similar findings from various by dissolving infected tissues, wound disinfection by experiments were also obtained by Yasin, [5]. Larvae killing e bacteria and stimulating e wound healing. For mostly tend to pupariate near e food sources. Goday et wound repair and regeneration maggots eraphy have al. [7] Pointed out at in most of e cases maggots been used for various purposes as reported by Sherman preferred food materials for pupation but Ghafoor et al. [0]. This erapy has also been investigated in United [7] proved at decayed fruits were best residing site for State for wound healing and repairs in horses since 2003. pupation. In e batches of 50 (October) soil (77 5d ) General behavior and status in various animals of attracted maximum number of pre-pupae on e contrary blow and flesh flies had been studied by Shah and Yesin, [8] declared at soil was e moderately attractive Sakhawat [6]; Wells and Kurahashi []; Aguiar and site for pupation. Milward [2]; and Tachibana and Numata, [3]. Various In e present research during e mon of environmental factors including temperature and December, batches 0, feaers and cotton (70 5d ) photoperiods are seriously involved in e dispersion of attracted same and maximum number of pre-pupae and e post feeding larvae. Dispersion and pupation site bread crumb proved least attractive site for pupation. preference was studied by Ghafoor et al. [4]. Life history, Ghafoor et al. [7] found fresh leaves were e best behaviour and pupation site preferences of bo flies were materials which attracted maximum number of maggots. In observed by various researchers [5, 6, 4]. Several species e batches of 50 feaers proved best preference of blowflies and flesh flies transport a variety of material for e residing of maggots but most of e gastrointestinal paogens and are carrier of number of auors [6, 6, 8] studied at bark was e most diseases. Isolation and detection paogenic H5N- preferred material for e pupation. In e resent study, Avian Influenza a virus from blowflies were collected from pre-pupae moderately preferred e cracked bark for eir vicinity of infected poultry farm in Kyoto, Japan in 2004. pupation. Ghafoor et al. [7] investigated at folded Same species were used in e current experiment to cloes were e least attractive pupation material but dry evaluate e responses for pupation sites preference by leaves and newspapers were e moderate attractive sites applying various pupation substrates. Insects play much for pupation. It is a need of time to explore e general important role in determining e postmortem, interval and behavioral, dispersal patterns and various aspects of post location of dea [3]. Analysis of environmental factors feeding larvae of blow and flesh flies. In future eminent particularly photoperiod and temperature can be helpful researchers should be encouraged to investigate about in searching for dispersing of larvae around cadavers [5]. e medical importance of Calliphorid and Sarcophagide Larval and pupal behaviors of blow and flesh flies were flies.
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