protocol green booklet #2 another success story

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Another Success Story! This is a real life example of a real problem solved with the new green treatment formula of Protocol Green! CALIFORNIA HUD APARTMENTS INFESTATION SINCE 2009 NINE PEST CONTROL COMPANIES FAILED TO ERADICATE AND PREVENT THE PROBLEM! CALL TODAY! TREATED 2014 NOW BED BUG FREE! Client will now spray with Protocol Green "Prevent" every vacancy to stay Muti-Insect FREE! Protocol Green’s one time initial application provides a safe immediate kill and future protection and eradication of reintroduced bed bugs in all of its life cycles for 12 months! Make a happy Customer For life!

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Page 1: Protocol Green Booklet #2 Another Success Story

Another Success Story! This is a real life example of a real problem solved with the new green treatment formula of Protocol Green!

• CALIFORNIA HUD

APARTMENTS • INFESTATION SINCE

2009

• NINE PEST CONTROL COMPANIES FAILED TO ERADICATE AND PREVENT THE PROBLEM!

CALL TODAY!

TREATED 2014 NOW BED BUG FREE!

Client will now spray with Protocol Green

"Prevent" every vacancy to stay

Muti-Insect FREE!

Protocol Green’s one time initial application provides a safe immediate kill and future protection and eradication of reintroduced bed bugs in all of its life cycles for 12 months!

Make a happy Customer

For life!

Page 2: Protocol Green Booklet #2 Another Success Story

                   Protocol  Green  Tes monials

Sometimes  satisfied  customers  say  it  best

"I had tried other products to kill bed bugs and spent a lot of time and money. Protocol Green really works." Dave  M  Hospitality  General  Manager,  California

"11  months  later  s ll  no  material  re-infesta on." Jeff  B. Government  Agency “My  customer  had  several  previous  pest  control  professionals,  including    “name  brands”  try  to  kill  bed  bugs  in  a  very  heavy  infesta on  from  their  shelter  facility.  My  customer  said  I  was  a  hero,  I  now  use  Protocol  Green  exclusively.”                        Dennis  O. Owner/Operator  Pest  Control  Company,  Florida. "Protocol Green kills bed bugs and I told all my friends about it. Homemaker,  Pennsylvania "Saved me time and money. Everybody should know about Protocol Green" Jay  P. Hospitality  Owner,  Vir-­‐ginia  

“We  have  used  the  product  in  homes,  hotel  rooms  and  apartments  with  equal  success.    The  fact  that  the  product  is green , nontoxic and effective makes it an ideal choice for a broad spectrum of application without worrying about potential harm to humans, animals or the environment. The year-long residual reduces callback and gives our clients peace of mind. PMP  Management  Services  Owner  Franchisee,  Florida

“We  had  just  taken  over  two  buildings  with  a  bad  case  of  bed  bugs.  We  went  in  with  Protocol  Green  formula  and  I am happy to say these accounts are bed bug and insect free a year later. What a great tool for our arsenal." PMP  Owner  New  York

"I tried Protocol Green on some bed bug-infested apartments and within two weeks all activity had ceased. I am now  in  the  bed  bug  business.”        PMP  Owner    TN

                     Www.technicide.com      800.950-5866

Page 3: Protocol Green Booklet #2 Another Success Story

YOUR BUSINESS IS ABOUT TO EXPLODE !

PROTOCOL GREEN FOR ELIMINATION AND RESIDUAL PROTECTION

APARTMENTS  

HOTEL/MOTEL

TIME  SHARE  RESORTS

CRUISE  LINES

AIRLINES

PUBLIC  TRANSPORATION

SHIPPING  CONTAINERS

THEATERS

MOVING  VANS

HOSPITALS  

NURSING  HOMES

COLLEGE  DORMS

GOVERNMENT  FACILITIES

MILITARY  BARRACKS

RETAIL—COMMERCIAL

SINGLE  FAMILY  HOMES

DON’T  LET  YOUR  COMPETITION  GAIN  YOUR  MARKET  SHARE      WITH  THE  NEW  SECRET  WEAPON  AGAINST  BED  BUGS  AND  MORE! FIRST  EVER,  NON  TOXIC  PESTICIDE  THAT  WORKS  FASTER,  LASTS  LONGER  THAN  ANY  PRODUCT  ON  THE  MARKET  TODAY  

Page 4: Protocol Green Booklet #2 Another Success Story

I. Report of Feb. 9, 2009 - The Questions to be Resolved where:

A)

B) When used according to label directions is the product lethal to the insect pests specified?

C) Does the product provide illumination under black light where applied?

Samples were sprayed and left to dry in ambient temperature “…the following was observed with 100 lumen 350 nm black light in a darkened room: each sample of painted plywood, raw plywood, ceramic tile, carpet, cotton fabric, and glass exhibited a glowing of the residual left behind on the test substrates.”

II. Report of March 14th, 2009 – The Questions to be Resolved were:

A)

“ It is nontoxic to mammals. It is not harmful to the environment. It is lethal as to the identified target insects.”

B) To what specific insect pests does this Certification apply?

Protocol GreenProtocol Green » Efficacy Studies Summaries:

technicide.com / 800-950-5866 Page 1 of 4

Overview – This summary is drawn from five (5) studies that first began in February 2009 with the last being completed in January 2012. Three of the five studies are lab tests. Two studies are field tests conducted at two (2) separate locations over a twelve-month period with one of the two studies a follow-up test at one of the field locations to test residual lethality where Protocol Green formula had been applied one year previous.

Each study attempted to determine the truth of a specific hypothesis and was intended to answer certain questions about the product’s proper regulatory classification and performance capability. All of the studies were independently performed by the American Academy of Entomological Science and each is certified by the Academy’s director, Dr. Jeffrey K. Brown. Ph.D., R.P.E., B.C.E. (see more at the end of the report).

Efficacy is understood to mean that the products produce the results provided for in their label when applied according to label directions and that it is at the same time in conformity to all applicable US Environmental Protection Agency regulations.

I have combined each of these elements according to the instructions and formula that have been provided. These elements retain their chemical integrity in combination and do not combine to form any subsequent element that would not be Generally Recognized As Safe (GRAS). Or that does not comport with USDA 21 CFR 178.3400, Parts 182 and 184 as exempt food grade proteins.”

A copy of any report will be provided upon request. When asking for a copy of the report, please indicate the report by its “date” and provide your email address.

" elements are exempt under EPA regulation commonly referred to as 25(b) and are found on a list of elements commonly referred to as the ‘4A List’ and are Generally Recognized As Safe (GRAS).”

This Report is limited in its scope to the target insects which are identified as Cimex lectularius (Bed Bug), Periplaneta americana (American Cockroach), Blatella germanica (German cockroach), Ctenoceplhalis felis (fleas), Diplopoda (Millepede), Chiopoda (Centipede), Lepisma (Silversfish), and Formicidae (Ants).”

Will the Academy Certify that the product is nontoxic to mammals, of no harm to the environment and lethal to the identified target insects?”

Following label directions it was applied to “…wild strains of house flies, Musca domestica, peri-domestic American cockroaches, Periplaneta americana, and ants of the family Formicidae, of which we observed 100% mortality within 2 minutes with house flies, within 5-10 minutes with American cockroaches depending on life stage (adult females took longer to cause mortality) and within 5 minutes with ants. Bed Bugs, Cimex lectularius, 100% mortality within 5-10 minutes depending on life stage.”

Do the products comply with applicable FIFRA (Federal Insecticide, Fungicide and Rodenticide Act) regulation’s Section 25(b) being of “minimal risk inert ingredients as referred to as ‘4A’ under Pesticide Registration Notice 2000- 6”? In addition, is the active ingredient compliant with 40 CFR section 152.25(g) of Title 40, a Generally Recognized As Safe (GRAS) ingredient. * In order that these questions are answered, Technicide (Protocol Green) provided quantities of all active and inactive ingredients for analysis. The ingredients were then combined according to proprietary formula and again analyzed to determine if in combination the formed element remained in conformity. The test concluded that it did.

TM Efficacy Data

Page 5: Protocol Green Booklet #2 Another Success Story

C) Is the Material Safety Data Sheet (MSDS) form accurate?

“ …all ingredients in your product are safe and non-toxic to humans and other animals as explained in your Material Safety Data Sheet (MSDS).”

D) Is the product qualified as “Exempt” under current EPA regulations and will therefore not require registration?

“ In our estimation your product will not need EPA registration as all of your components are presently categorized as exempt and there are no restrictions as to means of transport under DOT regulations.”

E) Upon what standards does the Academy base its Certification?

“ The Academy’s Certification, this report and our conclusions are based upon (1) theory and technique that can or has been tested; (2) theory and technique that has been subjected to peer review; (3) a known or potential rate or error; (4) the existence and maintenance of standards controlling the technique’s operation; and (5) general acceptance in the scientific community. As you have provided the raw components of your product together with your proprietary instructions as to the method and conditions of combination of components whose chemistry is already familiar to us, we have completed our evaluation according those portions of the GLP’s as are appropriate and necessary.”

III. Report of July 23rd, 2010 – The Question to be Resolved was:

A) Because 50% of infestation is in the form of hidden and untreated eggs what is the effect of residual lethality on emergent nymphs and instars?

“ Nymphs mortality was observed at between one (1) to two (2) hours; first and second instar nymph mortality at sixty (60) % within 1.5 hours after exposure. Fifth instar nymph and adult mortality were observed between four (4) to six (6) hours after exposure; fifty (50) % within 4.5 hours.”

IV. Report of November 10th, 2011 – Field Study of One (1) Year

This report consisted of a single treatment at each of two (2) public facilities, one a public museum, designated “Facility A” and the other a private commercially operated hospitality facility, designated “Facility B.” Both facilities had been previously treated with conventional toxins by licensed pest control operators.

Control was not achieved by these standard methods. Facility A continued to be infested with Dermestidae Anthrenus spp., carpet beetles and Facility B continued to be infested with Cimex lectularius, bed bugs. Approximately six months after these unsuccessful applications the Academy oversaw the testing reported.Application was made according to label directions and the sites were monitored for a period of one (1) year with periodic monthly reporting.

The Questions to be Resolved were:A) Did application provide control?

“ Immediately following application at Facility ‘B’ the bed bugs were observed for knockdown. Follow up observations and collections were completed at intervals of 0.5 hours, 1.0 hours, 24 hours, 48 hours, one month and every subsequent month for the next twelve (12) months.”

B) Did the U.V. markers continue to be observable?

“ After 1.0 hours UV marker was observed with 350 nm ultra violet black light.” UV marker continued to be observed with 350 nm ultra-black light.”

C) Were the Facilities subject to continued invasion?

“ The noted periodic inspections of the facility (Facility A) and insect traps revealed that some adults continued to ‘invade’ the facility as it was not sealed to the outside.” “Facility B continued in commercial operation and was subject to periodic use by guests and was serviced according to the regular daily housekeeping services.”

D) Was there evidence of residual lethality?

“ Carcasses gave evidence of residual kill at each subsequent periodic monthly inspection.”

Immediately following application at Facility ‘A’ the carpet beetles were observed for cleanout. Follow up observations and collections were completed at intervals of 0.5 hours, 1.0 hours, 24 hours, 48 hours, one month and every subsequent month for the next twelve (12) months.”

Protocol Green was effective against carpet beetles, Anthrenus spp., and bed bugs, Cimex lectularius. When spraying bed bugs with the test substance Protocol Green clean out was completed as to all visually observable bed bugs in the treated area. Protocol Green was also a successful killing agent to the extent that the treated area remained materially free of carpet beetles and bed bugs for the subsequent eleven (11) months.”

technicide.com / 800-950-5866Page 2 of 4

Page 6: Protocol Green Booklet #2 Another Success Story

E) Was there material re-infestation?

“ No material re-infestation has been revealed in any subsequent periodic monthly inspection.”

“ There were no reports by guests at field Facility ‘B’ of any evidence of infestation in the periods between the 24 hour intervals of periodic inspection in treated areas for twelve (12) months.”

“A

F) Was any odor present in any treated area, did any stains appear or were there any signs that application had caused damage or spotting?

“ No odor was present, no stains or damage to any materials was observed.”

G) In how many rooms at Facility B was the product tested and were records kept of the periodic inspections of those rooms?

V. Report of January 30th, 2012 - Field Study of One (1) Year Follow up

The Question to be Resolved is:

A) Is there residual efficacy twelve (12) months after application

upon re-introduced bed bugs?

Rooms at Facility ‘B’, the commercially hotel/motel facility were divided into two groups, Group A and Group B. “ Group ‘A’ consists of rooms previously treated… Group ‘B’ are the untreated rooms, not part of the prior project and with no prior report of infestation.”

“ Rooms in both Groups remained in continuous service and have subjected to regular housekeeping and inspection, including periodic bed bug dog inspection”

“ Room 1 was randomly chosen from Group ‘A,’ the previously treated rooms.”

Studies Performed By:American Academy of Entomological SciencesDr. Jeffrey K. Brown, Ph.D., R.P.E., B.C.E.P. O. Box 2503 Ridgeland, MS 39158 www.entomologyacademy.com

Jeffrey Brown received his B.S. in Vertebrate Zoology with a minor Wildlife Disease and Zoo Medicine and Ph.D. in Medical Entomology (minor in public health) from the University of California, Davis and his M.B.A. from Central Michigan University.

Dr. Brown spent three years as executive Director of the American Sanitation Institute where he directed a 30-person work force of sanitarians that addressed public health issues, problems and solutions to bring food manufacturers into compliance in the food processing industry.

Dr. Brown has completed over 1,000 consults; and developed the only in house insect and rodent clean out system allowed by the FDA in Atlanta, Georgia and defended a $350 million company which has been seized by Federal Marshals.

He moved to Wisconsin where he was the Director of Technical Development for S. C. Johnson Wax. With the help of his team they developed and launched the Vector Electronic Fly Trap that spawned a new market segment for Johnson Wax.

Dr. Brown is currently Bureau Director and State Medical Entomologist in the Health Department for the State of Mississippi and Director General of the American Academy of Entomological Sciences. The academy’s team of scientists performs contract research on insect species of public health importance.

Dr. Brown has been brought into Cairo, Egypt to address the Government, Military, Hospitality and Food industries on bed bug biology, ecology, and most importantly control. He will be presenting to the government of the United Arab Emirates in Dubai and Abu Dhabi and in Saudi Arabia, Morocco, and Turkey on bed bug control in the spring of 2013. Dr. Brown is a leading proponent of safe, non-toxic, green but effective bed bug and other insect control methods and the Academy has just finished donating their time to treat the Homeless Shelters for bed bugs in the State of Mississippi using these safe and non-toxic methods.

As to Facility ‘A’, Protocol Green was immediately effective against carpet beetles, Anthrenus spp., and remained effective over the next twelve (12) months to the extent that no further material infestation was present.”

s to Facility ‘B’, Protocol Green was immediately effective against bed bugs, Cimex lectularius, and remained effective over the next twelve (12) months to the extent that no further material infestation was present.”

As to Room 1 from Group ‘A’, having been previously treated with Protocol Green and where UV light confirmed sufficient residual element still in place and having had no intervening treatments or material occurrence of infestation Protocol Green was proven effective against re-introduced bed bugs, Cimex lectularius, where treatment had occurred more than twelve (12) months previous.” (See Table 2 attached to this Summary)

technicide.com / 800-950-5866Page 3 of 4

Twenty (20) guest rooms were selected. Of these eleven (11) were known to have current infestation and nine (9) uninfested guest rooms shared common walls with the infested rooms. Table 1 (Hospitality Facility) provides a twelve (12) month record as regards the treatment and inspection of these rooms. Cimi-Shield was used as to infested rooms and PG-Protect, the preventive formula, a derivative of Protocol Green, was used as to the neighboring but infested rooms. (Table 1 is attached to this Summary)

The failure to adequately seal Facility ‘A’ permits ingress of carpet beetles where the residual agent of Protocol Green as seen under black light has continued to provide evidence of its lethality as carcasses are observed and most heavily concentrated in areas closest to poorly sealed or unsealed thresholds and around door casings.”

Page 7: Protocol Green Booklet #2 Another Success Story

Table 1 (Hospitality Facility)Monthly periodic on site monitoring of Bug Dome traps and once a year inspection with trained bed bug dogs physical inspection provide the following:

L = living M = moribund0 = no evidence

Results: Mortality was observed and recorded for the treated and control Room 1 in the following table.

MortalityGroup ‘A’ Room 1 TreatedHour 1 2 3 4 5 6 7 8 9 10 11 12 13 0 0 0 0 3* 0 0 0 4 3 2 0 1

Group ‘B’ Room 2 Control 1** 0 0 0 0 0 0 0 0 0 0 0 0

*Three (3) nymphs were dead** We are assuming that the researcher damaged the bed bug with forceps while removing from the travel container

before placing in the arena.

Table 2: Mortality of treated carpeting with Protocol Green on bed bugs one (1) year prior to the test versus control.

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Room # Product Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov1 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 02 PG-Protect 0 0 0 0 M 0 0 0 0 0 03 PG-Clean-Out M 0 0 M 0 0 0 0 0 0 04 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 05 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 06 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 07 PG-Protect 0 M 0 0 0 0 0 0 0 0 08 PG-Protect 0 0 0 0 0 0 0 0 0 0 09 PG-Protect 0 0 0 0 0 0 0 0 0 0 010 PG-Protect 0 0 0 0 0 0 0 0 0 0 011 PG-Protect 0 0 0 0 0 0 0 0 0 0 012 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 013 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 014 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 015 PG-Clean-Out M 0 0 0 0 0 M 0 0 0 016 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 017 PG-Protect 0 M 0 0 0 0 0 0 0 0 018 PG-Protect 0 0 0 0 0 0 0 0 0 0 019 PG-Protect 0 0 0 0 0 0 0 0 0 0 020 PG-Clean-Out M 0 0 0 0 0 0 0 0 0 0

ConclusionsThe test substance Protocol Green Clean-Out was complete as to all visually observable bed bugs in the test area.Protocol Green Knock-Out was an effective preventative against bed bugs, Cimex lectularius, twelve (12) months after ap-plication per label directions.

Page 8: Protocol Green Booklet #2 Another Success Story

THE RESEARCH REPORT: PROTOCOL GREEN BED BUG NON-TOXIC ELIMINATOR & RESIDUAL

There are many choices in the market today to battle bed bugs. In addition to applying traditional insecticides, extreme temperature treatments have grown in popularity. In hospitals, outpatient centers, nursing homes, dormitories, schools and hotel rooms where sensitive health and chemical exposure are concerns, temperature treatments are a common choice. They are effective, however, the main drawback is they leave no residual and these types of sites are areas where incoming pressure is a constant and more of an issue than infestation. Another important reason to use a residual product is because experts believe as much as 60 % of an infestation consists of the bed bug eggs that will hatch later.

Is it possible to develop a non-toxic chemical that is lethal to bed bugs, yet leaves a residual that is harmless to humans and other mammals? The American Academy of Entomological Sciences was approached by a private laboratory to test a product for its effectiveness in killing bed bugs, Cimex lectularius), in a field situation with residual potential. Generally, we do not test any product without knowing the active and inert components and the mammalian toxicity.

Dr. Jeffrey Brown signed a confidential disclosure and received the list of ingredients. When Brown saw that the active was an EPA-exempt     25(b)     component,   “soybean   oil,”   he  contacted the laboratory  and  explained  to  management  that,   in  the  team’s    professional    opinion,    this

The product we tested is commercially available from Technicide as Protocol Green from www.technicide.com and its distributors. It is available in two strengths: CLEAN OUT (high concentrate), and PREVENT (low-concentration residual).

   “active”  was    not likely to be effective on insects and we were concerned that they would be wasting their money. They insisted and wanted to explain the manufacturing process and what they do with soybean oil to achieve this unique response. Having had the benefit of their explanation and learning of the background  for  the  product’s  development, the team agreed to do a small lab test with minimal cost.

At first we did not believe that the product would be effective in killing bed bugs. However, once the tests were completed and more was learned about mode of action, the results spoke for themselves. The active is a proprietary protein complex initially engineered from soy-bean oil involving thousands of proteins and their specific ratios. This active bio agen is found commercially in Protocol Green.

Research Field Testing Data Report

Page 9: Protocol Green Booklet #2 Another Success Story

SITE DESCRIPTION. The site was a commercial hospitality facility offering overnight and extended stay, single-room accommodations, meeting facilities and other guest services to the public. It is professionally managed with on-site and regional support supervision. The structure is four stories of single rooms, each about 300 square feet. Each room is carpeted, has one to two queen-sized beds with box springs and mattresses, affixed headboards and a chest of drawers. Bearing walls are placed one on top of another without concrete separation. All but end rooms share at least two common walls, one on each side, ceiling and floor joists, except for bottom and top floors which have but one adjacent room above or below, respectively. End rooms share one common wall. Electrical lines are placed in the wall voids and run from room to room.

All treated rooms were fitted with bug dome traps and inspected monthly. There was also an inspection with a trained bed bug dog once each year. In the ensuing 12 months, no live bed bugs were found in the treated rooms. In the 11 rooms that were initially treated for active infestations, all showed dead bugs one month after treatment. Only two rooms had an additional occurrence in the 12-month period where dead bed bugs were found. In the nine rooms treated preventively, only three occurrences were observed during the same period as dead bed bugs were found. The last four months of the observation period showed no evidence of bed bugs in any of the treated rooms.

In these initial steps we had proof it worked to kill by direct contact, but the question of residual efficacy remained. The residual efficacy testing project required treating a site with current bed bug activity with a direct spray formulation (prepared in accordance with label directions), and careful periodic monthly evaluations for 12 months after the initial treatment.

Rooms are serviced daily by housekeeping with standard hospitality-style wheeled service carts that are pushed from room to room. Carpets are vacuumed with a common machine and were not shampooed during this test period. Soiled bed clothing, towels, and wash clothes are gathered daily from each room, placed in common service carts and moved from room to room until filled. They are then moved to an on-site facility and laundered.

INITIAL TREATMENT. Packets of super concentrated material in two formulations (CLEAN OUT and PREVENT only) were prepared according to label directions in clean 1-gallon hand pump spray canisters. It should also be noted that the sprayer canisters, hoses and nozzles must be completely free of any surfactant (e.g., soap), other chemicals or residues. This product cannot be combined with other 3 to 4 feet per second. This provided for an application rate of about 1 gallon of finished product per 1,200 square feet of surface area and included application to all fabrics and textiles. Treatment included the spraying of all carpeting, upholstered furniture, all sides of box springs, all sides of the mattresses and window coverings, and bed frames including plastic snaps. Headboard and headboard support were removed and these areas were sprayed with a single pass at a distance of approximately 24 to 30 inches moving at about 3 to 4 feet per second. No furnishings were removed. A single pass of all cracks, crevices, junction points and baseboards where carpeting met walls were treated with a spray at a distance of about 24 inches at a rate of approximately 3 to 4 feet per second.

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Although the subsequently found dead bed bugs could be attributed to the product’s   residual  effect,  more conclusive data was desired — we still wanted to know how effective the residual was. After twelve months, fresh bed bugs were brought in from outside the site. In this project we chose one room randomly that had been previously treated and a room (for our control) that had never been treated. The treated room had no observed odor, stains or damage. It was vacuumed and darkened in order to inspect the residual condition with a 350 nm UV black light (the greater the lumens, the easier    to    observe    the  “sparkle”  or  “glow”).  This  was  possible as the formula contains a UV reflective element so as to allow for confirmation of application on inspection. At 6 to 8 inches, it was apparent through visual inspection where product had been originally applied on textile surfaces.

SO HOW DOES THIS WORK? We have been asked several times about the mode of action of the proteins, and this is our understanding — the protein complexes uncouple electron transfers and increase transmembrane leak of the protons in the membrane of the waxy layer of the insect. This is similar to standard chemical uncoupling described in organic chemistry. In plain English: the waxy layer is breaking down (in some cases rapidly), creating a hole. Because   our residual   affects the waxy layer, rather than depending on getting into a receptor site, the insect cannot become resistant to the active protein agent and invites further investigation for application on other similar insect species.

This field study clearly indicates that the residual was effective 12 months after the initial application. The   project   ended   January   2012.   The   “clean out ”   formula   can   be   used   to   obtain   control   over   an infested area. The preventive formulation can be used in areas where no infestation is wanted. It can also be used as an appropriate  “top  dressing”  following an extreme temperature treatment, either heat or freezing (Cryonite), providing residual protection for at least a year afterward, even with regular vacuuming. It is important to note that regular sanitation and maintenance of this site contributed to the success of the treatment.

In the treated room, 10 adult and 3 nymphs were placed. The nymphs were all dead by hour 4 and the adults were all dead by hour 13. In the control room, an identical setup with 12 adults was placed and observed at the same time. There were no deaths except for three that were presumably injured in transfer from travel container to observation arena. At the end of the observation (hour 13), the control group was anesthetized with CO2, placed in the upright aquarium, allowed to revive, and sprayed    with    the    “Clean Out”  formulation.  Mortality  of  all  bed  bugs  was  observed  within  25  seconds.

The bed bugs were placed on the carpeting near the head of the bed under an inverted arena of a new plastic 10-gallon aquarium with the bottom inch of the aquarium treated with petroleum jelly to prevent the bed bugs from escaping. Hourly checks were made in the evening hours to avoid disturbance by guests or housekeeping and to simulate the nocturnal cycle of the insects. Light was used only at the time of inspections and then with a flashlight. Observations were recorded until all insects were dead or moribund. Instead of introducing CO2 gas from a cylinder, the observer would slowly breathe into the inverted glass arena using a flexible tube for five breathes (about 10 seconds) to induce movement of the bed bugs across the carpeting.

Page 11: Protocol Green Booklet #2 Another Success Story

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