Jessica Gross's blog

Y-tube bioassay with beta-Phellandrene

This experiment was set up after dark, approximately 6 pm, when the beetles are most active.

50ul of beta-Phellandrene were put inside of a heat-sealed plastic straw, with two holes poked into the straw for release of chemical.

The air was set at 3.5 liters per minute. If the CRB did not make a decision at 2 minutes, that subject was taken out of the tube.

20 CRB were used:

1) Jar# 0930, female: at 1:54 min --> air

2) #1471, male: at 2:09 min --> lure

3) # 0154, female: at 4:17 min --> lure

4) # 1349, male: at 15 sec --> air

5) # 0772, male: at 3:21 min -->air

6)# 1172, male: at 17 sec --> air

7)# 0313, male: at 2 min no decision

8) # 0396, female: at 45 sec --> air

9)#1242, male: at 4:12 min --> air

10) # 1434, female: at 5 min --> lure

Cleaned y-tube with Acetone, switched lure sides

11) #1230, female: at 2:18 min -->air

12) # 1282, female: at 2:43 min --> air

13) # 0250, male: at 2:20 min --> air

14) # 1539, male: at 2:20 min --> lure

15) #1226, female: at 2:16 min --> air

16)#1227, female: at 2 min no decision

17) # 1234, male: at 2 min no decision

18) # 1241, female: at 55 sec --> lure

19) #0705, female: at 1:24 min --> air

20)# 0050, female: at 3:30 min --air

Current GMF experiment

On 12/16 we set out to test the efficacy of the entomopathogenic fungus GMF (Green Muscardine Fungus) on third instar larvae of the CRB.

Six larvae were used as a control and placed in mason jars filled with soil and debris from a compost pile found in the field. 2 tablespoons of GMF were then applied to the 1 cubic meter sized compost pile, and the material was stirred up to evenly distribute the spores. This time we took samples from the pile and added the material to the 6 remaining larvae.

There will be a follow up on the experiment every 4 days.

On 12/30/13 all control and treatment larvae were still alive. 20 ml of water were added to each jar to keep the contents moist. So far no GMF is visible.

Metarhizium bioassay 4

On 01/09/12 four soil samples from four different GMF sinks (Ada, Oka, Fujita 1&2) were taken and placed into glass jars containing healthy CRB larvae to see if they would get infected by the GMF growing in the soil. In total, six 1sts, 7 2nds and three 3rds were placed into separate jars. Also, all female beetles that are found in the GMF sinks are now being collected and brought in to test in this experiment.

Ada jar numbers:

0026: 1st instar

0036: 2nd instar

0033: 3rd instar

0037: 1st instar

Oka jar numbers: 

0038: 1st

0024: 2nd

0039: 3rd

0678: 2nd

Fujita 1:

0040: 1st

0031: 2nd

0041: 3rd

0682: 1st

Fujita 2:

0025:1st

0032: 2nd

0043: 2nd

0028: 2nd

Adult CRB jars:

1029

0045

One week later, the jars were checked.

Ada

0026: 1st was dead and disintegrated

0026: the 2nd was near death, not moving and turning hard

0033: 3rd still alive and active, but with black fluid coming out of its mouth

0037: 1st dead and green

Oka: 0038 alive 

0024: alive

0678: alive

0039: alive with black fluid coming from mouth

F1

0040: dead with green/white coloring

0031: alive with black fluid coming from mouth

0041: alive

0682: alive

F2

0025: dead and disintegrated

0032: alive

0043: alive

0028: alive

1029: alive

0045: alive

On 01/25/12 all jars were checked again and all the larvae were dead and covered with green hyphae. The adult beetles were still alive.

What is a GMF sink?

A GMF (Green Muscardine Fungus / Metharhizium anisopliae) sink is a method of attracting and killing Coconut Rhinoceros Beetles in their various life stages by means of building a ground trap. This ground trap consists of four coconut logs arranged as a squareshaped confinement on the ground, with either chipped wood or decaying coconut debris and soil in the center. The coconut logs and soil attract the beetles to the trap, where they breed and lay their eggs. Additionally, a pheromone lure is added to the trap to ensure that a large amount of beetles is attracted to the sink. The soil/debris in the center has been treated with the Green Muscardine Fungus, usually approximately 350 g, which leads to the death of the CRB within 14 to 21 days. In the time that the larvae and or adult beetles are still alive, they are capable of infecting other healthy beetles with the fungus as well. The sinks are then covered with coconut fronds to prevent UVA/UVB rays from damaging the growth of GMF.  In order for GMF to grow well, it needs warm temperatures and high humidity. If this is provided, the fungus can remain active in the soil for several months up to one year. We are currently inspecting these sinks every two weeks, re-applying GMF to them and recording the number of beetles and larvae (dead and alive) found inside the traps.

GMF sinks

On 12/20/2011, these GMF-filled sinks were made:

1) South of Hyatt Hotel: 220 g GMF. GPS coordinates: 13.51225, 144.80334

2) Fujita:  200 g GMF. 13.50908, 144.80225

3) Fujita: 410 g GMF. 13.50894, 144.80080

4) Ada: 220 g GMF. 13.49771, 144.78444

5) Paseo de Oro: 476.5 g GMF. 13.49943, 144.78513

 

On 01/10/12 further GMF sinks were made:

6) Wusstig, Yigo: 400 g GMF. 13.53361, 144.87030

7) Maina: 250 g GMF. 13.46790, 144. 73862

8) Sinajana: 250 g GMF. 13.46317, 144.75638

9) Toto, Bordallo's property: 500 g GMF,  13.458486, 144.769771

10): Nimitz Hill: 400 g GMF, 13.464485, 144.723542

13) Mogfog 1: 400 g GMF, 13.50681, 144.85307

14) Mogfog2: 400g GMF, 13.50804, 144.866792

15) Leo Palace : 400 g GMF, 13.41553, 144.73307

16) Asan Baseballfield: 500 g GMF, 13.47090, 144.71215

17) Piti firing range:500 g GMF, 13.44300, 144.68849

18)Talafofo Golf Course dump site: 451 g GMF. 13.36103, 144.73584

19) Our Lady of Peace Cemetery: 300 g GMF. 13.37595, 144.73471

20) Ukudu: 500 g GMF,  13.525689, 144.828696

21) Leo Palace Rd: 500 g

22)Palantat 1: 500 g

23)Palantat 2: 500 g

24Tanguisson 1: 500 g

25) Tanguisson 2: 500 g

(26) Urunao: 500 g,  13.627697, 144.840276

(27) Ritidian: 500 g,  13.651073, 144.869392

(28) Talafofo Onward roadside: 500 g,  13.361883,144.735464

 

 

 

 

 

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