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Lab 4 Forensic Entomology - Coggle Diagram
Lab 4 Forensic Entomology
RESULTS
Article 2
Study all instar stages of larva of
Hypopygiopsis tumrasvini Kurahashi
Difference between the third instar stage with other instar stages due to the third instar larva’s cephalopharyngeal skeleton lacking the accessory sclerite ventral to the mouth hooks and its anterior spiracle composed of eighth papillae while the second instar larva’s have 11 papillae.
Third instar: A head region that lacks a head capsule, 3 thoracic segments and 8 abdominal segments.
Results for both larvae stages and adult stage
Article 1
Study third instar larvae of necrophagous Calliphoridae
Difference between the third instar stage with other instar stages with the presence of anterior spiracles and the number of slits of posterior spiracles
Third instar: Divided into bilobed pseudo cephalon, 3 thoracic segments, 7 abdominal segments and the anal division.
Results only regarding third instar larvae stage
Article 3
Study all instar stages of larva of
Lucilia porphyrina
First instar: 12 segments with the last abdominal segment being truncated.
No significant difference shown between second and third instar larvae. (Similar range number of papillae at anterior spiracle, sensory structures at the tip of the head similar)
Results on the larvae stages and pupa stage
INTRODUCTION
Similarity
All of the three article agrees that fly has its significant in the forensic entomology.
Differences
Article 1
The morphology of third instar of calliphoridae
11 species included in the article where the insects are mostly wide spread through Europe
Calliphora vicina, C. vomitoria, Chrysomya albiceps, Phormia regina, Protophormia terraenovae, Lucilia caesar, L. illustris, L. sericata, Cynomya mortuorum, Chrysomya megacephala, and Lucilia ampullacae
Article 2
Focus on Southeast Asia region, Chrysomya, Achoetandrus, Lucilia, Hemipyrellia, Hypopygiopsis have forensic importance
In Oriental region, five species of Hypopygiopsis are included which are Hypopygiopsis fumipennis Walker, Hypopygiopsis infumata (Bigot), Hypopygiopsis tumrasvini Kurahashi, Hypopygiopsis robusta Malloch, and H. violacea
Thailand
Article 3
Focus on Lucilia species in their immature stages
Thailand
Lucilia porphyrina, Lucilia cuprina, Lucilia papuensis Macquart, and Lucilia sinensis Aubertin
METHODOLOGY
Article 2
(a) Larvae
Firstly, anterior position was cuts from the middle of the mesothorax
Then cut the end of the eight abdominal segment to remove the posterior of the body
After the cut, soaked the larvae for 5 minutes in 10% of KOH
Few drops of Neo- Shigaral Permount for mounting was put on the slide after transferring the larvae on clean slide
The images of the larvae was identify by using light microscope under x10 and x40 magnification
Images was recorded with digital camera (Nikon)
(b) Adult males and females
Photograph taken by using digital camera (NIkon,Japan)
Next, their genitalia was being cut from the posterior end and put under dissection microscope
Soaked the posterior end in 70% of alcohol for few minutes and transferred to 10% of KOH for cleaning
The cleaning takes 24 hours for male and 12 hours for female
After that, 0.85% of normal saline was put on a glass slide for dissecting the flies
The posterior end was mounted with Neo Shigaral Permount and covered with coverslip
The slides were examined by using light microscope
Article 3
(a) Light microscope
Firstly, the larvae was put in a rearing tray
Then, larvae will be boiled for few minutes
Transferred to 10% of KOH for cleaning
After that, cut the larvae into two sections (Anterior end and posterior end)
The posterior and anterior part were mounted with Neo Shigaral Permount and covered with coverslip
The images of larvae identify under x4 and x40 magnification by using light microscope.
(b) Scanning electron microscopy
Firstly the larvae will be boiled for few minutes and stored in 70% of alcohol
The larvae washed with phosphate buffer (pH 7.4) while slightly shaking it for 5 minutes
2.5% glutaraldehyde mix with the phosphate buffer in 4 degree Celsius for 24 hours.
Cut the larvae into two sections (Anterior end and posterior end)
Larvae will be rinsed with phosphate buffer, twice for 10 minutes
Then, the larvae being postfixed with 1% osmium tetroxide for 1 to 2 days at room temperature.
The larvae will be dehydrate by using 30%, 50%, 70%, 80%, 90% and 100% alcohol for 12 hours.
After that, the larvae will be dehydrate by using acetone for 12 hours.
The larvae was being dry up and double-stick tape the larvae on aluminum stub coated with gold in sputter-coating apparatus.
Lastly, view the larvae under scanning electron microscope
Article 1
a. Larvae is soaked in hot water
b. Then it is stored in 80% of ethanol.
c. After the larvae are stored in the 80% of ethanol, it was soaked in 5% of KOH for 5 minutes.
d. Some of the particular fragments of the body were mounted in Hoyer's medium or dehydrated through 80%, 90% and 99.5% of ethanol and mounted in Euparal.
e. The slides mounted with two different slides, cephaloskeletons was mounted in concave slides and flat slides was for other morphological details.
f. Identification of the images was identify under digital Nikon 8400 camera that was mounted on Nikon Eclipse E200 microscope and Nikon SMZ1500 stereomicroscope.
DISCUSSION
Article 2
Third-instar larvae of
H. tumrasvini
is similar in morphology to that of
H. violacea
and
H. ligurriens
. However, accessory sclerite of the cephalopharyngeal skeleton is absent in
H. tumrasvini
but present in
H.violacea
and
H. ligurriens
.
For
H. tumrasvini
and
H.violacea
, the number of papillae on each anterior spiracle can distinguish them. For
H. tumrasvini
and
H. ligurriens
, the proximity of the pair of posterior spiracles to each other can distinguish them.
The spines between the prothorax and mesothorax of larvae of Luciliini are unique feature.
Article 3
Sculpture of the dorsal spines between prothorax and mesothorax displays unique features among Lucillini. (same with article 2)
Luciliini have larger size and apparent fewer spines than Hemipyrellia or Hypopygiopsis.