Port of Entry/Exit Pest Monitoring Program
FY22 HISC FUNDS PROJECT REPORTThis annual report includes the following components:
1. Project Overview
2. Report on activities in FY22
3. Economic Analysis by Donna Lee
4. Africanized bee report from UH Bee lab
Should you have any additional questions regarding the implementation of the Port of Entry/Exit
Pest Monitoring program, (formerly known as Māmalu Poepoe) in FY22, please do not hesitate
to contact Dr. Kaufman at Leyla.V.Kaufman@hawaii.gov.
1. Project Overview
The Port of Entry Pest Monitoring Program (formerly known as Māmalu Poepoe) was originallyconceived by representatives from multiple state agencies acting in their capacities as membersof the HISC, including the directors or designees from HIDOT, theDepartment of Health (DOH), the Hawaii Department of Agriculture (HDOA), and the
University of Hawaii (UH). These agencies recognized the following areas of shared interests
with regard to airport
facilities:• HIDOT seeks to understand the presence and impact of invasive species at airport
facilities that
may be detrimental to facility operation or user experience,• DOH seeks to improve its monitoring and research efforts regarding vectors of humandiseases atairports, primarily mosquitoes,• HDOA seeks to improve monitoring and research efforts regarding agricultural pests at
airports,
namely invasive ants, coconut rhinoceros beetle, and Africanized bees,• UH seeks to improve research on invasive species distribution and economic impacts,
and,• DLNR is the administrative host of the HISC, which is mandated to provide cabinet-level
coordination on invasive species issues.
HISC staff agreed to serve in a coordinating capacity for this project, including management of
an interagency project budget and the hiring of a temporary Project Coordinator via a staffing
partnership the UH Pacific Cooperative Studies Unit (PCSU) to finalize and implement the
Māmalu Poepoe plan.
Utilizing UH PCSU as a project staffing entity has allowed HISC to fund temporary positions
and partial full-time equivalences for the pilot project rather than establishing permanent civil
service positions. HDOT agreed to fund the program as a 5-year pilot project. The pilot funding
ended in March 2022.

2. Report on activities in FY22•Interview with project partners
HDOT pilot program funds expired in March 2022. During FY 22, the project
coordinator and CGAPS staff interviewed project partners about the need to continue
with the program. A battery of interview questions was generated for Federal partners(USDA, FWS), State partners (HDOA, HDOH, HDOT, UH), and RCUH project partners(ISCs managers, HAL manager). The responses were in favor for project continuationand for an expansion of efforts to seaports, as well as an expansion of invasive targets.

•

Program expansion
Given the overwhelming support from partner agencies, HISC staff worked on securing
funds to continue with the program. During FY 22 the project coordinator worked with
HISC/DLNR and CGAPS staff to send federal appropriation and Congressional Direct
Spending (CDS) requests to the different Congressional offices in Washington DC. HISC
staff also briefed local legislators on the program, which resulted in the introduction of abill. Federal and State requests had budgets for the continuation of efforts at airports andalso to expand efforts to seaports and expand the list of invasive targets. The CDS request
was approved, and the appropriation went to USDA APHIS. USDA APHIS channeled
the funds to DLNR via CAPS financial agreement. The program will expand efforts tothe following harbors: Honolulu, Nawiliwili, Kawaihae, and Hilo. The State
appropriation went to HDOA.

•

Working Group Meetings
Between August 2021 and July 2022, the program coordinator held 1 Questions andAnswers meeting with program partners (December 7th 2021) and two Working Group
meeting (August 3 2021 and April 21 2022).
The December Question and Answer meeting was attended by all the representatives
from the Invasive Species Committees, Hawaii ant lab, CRB response, UH Bee Lab. The
goal for the meeting was to standardize approaches among groups and work out any
issues with the NRDS database.
The working group meeting in August 2021 and April 2022 were attended by 38
participants and 35 representatives from DLNR, DOT, DOA, DOH, UH, CGAPS, CDC,
Tripler Medical, USFWS, as well as representatives from the different invasive species
committees and the Hawaii Ant Lab. These meetings provided updates on monitoring
efforts for mosquitoes, ants, Africanized bees and coconut rhinoceros beetle. It also
provided an update on the economic analysis done by UH economist.

4.1 INVASIVE ANTS
The program continues the collaboration with the Hawaii Ant Lab (HAL) and MISC for the
ant monitoring. During FY22 the program had two ant surveys at HNL and 3 surveys at ITO,

3 at KOA and 1 at OGG. No new ants to the state were reported at the different facilities.
Positives for little fire ant (LFA) were recorded at ITO and KOA airports.
2.2 AFRICANIZED HONEYBEES (AHB)
The University of Hawaii (UH) Bee Lab continues to provide guidance to the program. The
UH Bee Lab conducts the DNA analysis and swarm trap processing for swarms interceptedat HNL. Table 1 shows the number of swarm interceptions at airport facilities. All swarms
intercepted to data have tested negative to Africanized genes.
Table 1. Number of swarms intercepted at all airports from August 2021 and July 2022
Airport facility# swarm interceptions
Results of DNA analysis
Lihue1
Honolulu
8
Negative
Kahului
0
Hilo
4
Negative
Kona
1
Negative
Total
14
The program coordinator held 3 action plan meetings for AHB between August 2021 andJuly 2022). The meetings were held in September 2021, November 2021 and February 2022.
In these meetings the group discussed issues like mandatory bee registration, issues regarding
the importation of bee semen, buffer zones around airports, trainings for response, outreachmaterials, among other issues.
2.3 COCONUT RHINOCEROS BEETLE
No beetles were intercepted during FY 2022. Traps are currently being checked every fourweeks. The CRB response team has seen increase in CRB detections around HNL. Annual
palm surveys were conducted at all airport facilities. During the palm surveys monitoringcrews inspect palms for possible CRB damage and count number of palms. Figure 1 shows
the clusters of palms at Daniel K Inouye International airport. All palms surveyed were freefrom CRB damage.

Figure 1. Palm survey inside the AOA at Daniel K Inouye International airport
2.4 MOSQUITOESNo new mosquito species were intercepted during monitoring surveys at airport facilities
during this fiscal year. The program continued providing monitoring equipment to offices.
The program helped with the response to an interception of a female Aedes aegypti (thisspecies is only known to occur in Hawaii island) at Honolulu Harbor in August 2021. The
program helped with communications with HDOT partners to gain access to additional
harbor sites. The program also provided monitoring traps to help with the response. No otherinterceptions were recorded.
2.5 BEST MANAGEMENT PRACTICES (BMPS) FOR AIRPORT STAFF AND
AIRPORT CONTRACTORS.
The program put together BMPs for all target species. The BMPs outline practices to befollowed to maintain and operate airport facilities in a way to minimize the risk of invasive
species establishment on airports properties. In FY22 the BMPs were updated to reflect thecurrent CRB interim rule and compliance agreement put in place by HDOA. The document
also got a review from HDOT harbors.

Economic rationale forcontinuing monitoring invasive
species at Hawaii airports
Benefits from Monitoring - By specie

Monitoring and rapid response cost - AHB
Monitoring

Rapid response

Monitoring andresponseOahu$15,473$7,056$22,530

Kauai$7,075$4,531$11,606

Molokai

$0

$0

$0

Maui

$7,527$4,173$11,700

Hilo$19,582$8,258$27,840

Kona$11,203$5,335$16,538All$60,861$29,354$90,214

Benefit from monitoring for AHB at airports
Monitoring,
rapid
response, andfollow-up*

Terminal Closures*
5Days per year
15

20

Cost at 6
airports$90,214$2,659,109$7,977,327$10,636,436

Benefit-Cost
Ratio

Cost

29:1

88:1

118:1

*Close one terminal at each airport during swarming

Interpretation of B:C = 29:1
 If an outbreak is likely to occur more than once every 29 years,
monitoring to prevent the outbreaks is economically warranted
 If an outbreak is expected to occur only once every 29 years,
monitoring to prevent the outbreak, benefits equal the costs
 If an outbreak is expected to occur fewer than once every 29 years,
monitoring to prevent the outbreak (at the current level) is not
economically warranted.

Benefit from monitoring for LFA at airports
Monitoring

Airport Gate Closure*

Mitigation*

Days per year
12

52

260

Cost at 6
airports$66,574$196,210$713,062

3,089,934$15,449,672

Benefit-Cost
Ratio

Cost

3:1

11:1

46:1

232:1

One gate at each airport is infested and closed for treatment
Mitigation treatment is in place of routine monitoring and response
Gate closure cost compared to monitoring cost

Benefit from monitoring for CRB at airports
Monitoring

Mitigation

Landscape Damage*

Trapping

Tree replacement

Cost at 6 airports$29,326$169,688$539,795

Benefit-Cost
Ratio

Cost

6:1

18:1

*13.4% of the palm trees will be damaged irreparably. Overall % based on tree replacement rates of 1%(Kona), 4% (Molokai) and 15% (remaining) airports
Mitigation instead of monitoring and routine response
Landscape damage compared cost of monitoringBenefit from monitoring for mosquitos at airports
Avoided loss in airport net
revenues due to active cases of
disease for 6-months

Monitoring

Response

Total

Cost

Cost

Cost

Per year

Per incident

Per incident
One yearNet revenues

Benefit-cost

Culex (or all)$519,137$190,635$709,772$15,318,260

22:1

Anopheles$519,137$69,478$588,615$15,318,260

26:1

Aedes

$519,137$11,319$530,456$15,318,260

29:1

Progress and Ongoing Needs in Hawaii

Africanized Honeybee Detection and Prevention
Report by E. M. Villalobos - UH Honeybee ProjectHistorical perspective of swarm trapping in Hawaii
In Hawaii, monitoring for Africanized Honeybees at airports began in 2017 with the use of swarm
traps and pheromone lures. Since then, swarm catching procedures were modified in the middle of2019 when the UH Honeybee Project began assisting with the genetic analysis of the captured
swarms. The UH bee team also began recording the amount of honeycomb accumulated and thebrood condition of the swarms when they were delivered for screening of their genetic status. Trap
occupancy is now checked every 2 weeks and the contents of the trap are examined for bee pests
and brood age, which is related to maturity of the colony. The presence of drone bees is recorded,since they represent a possible dispersal route of genetic material linked to the colony being
examined.
Swarms are typically composed of young bees and an older queen bee. Once they settle in a swarm
trap, the bees quickly begin to build comb. This process is energetically consuming, and the swarm
bees carry with them a belly full of honey to begin this process. Honeycomb needs to be created
before food storing and egg laying can begin. So, during the first few days the bees will spend mostof the time creating and excreting wax, which is then shaped into the cells that form thehoneycomb. Once the nest begins to take shape, collection and storage of nectar and pollen canstart and the queen is ready to initiate egg laying and produce a new generation of worker bees.
From that point on, the speed of nest development is somewhat variable, depending on the numberof worker bees that was part of the swarm, the availability of resources nearby, and the weather.
However, the duration of the larval stage of a bee, from egg to pupa, is relatively constant at 6days. The pupa takes about 12 days to transforms into the adult bee. When our team processes theswarm trap samples, we consider the presence of larvae and capped brood (which is indicative of
pupal development) to assess the age of the swarm.
In addition, the appearance of drone cells in the comb structure gives us another indication that thecolony has been there for a while. Male bees, or drones, are produced only when a colony is strong
enough, which is likely correlated with age of the colony. Thus, swarms do not often produce
drones during colony establishment phase. Drones also take longer than worker bees to develop
because of their larger body size, 24 days as opposed to 18 by worker bees, so their presence
denotes an older colony.
The recommendation to have bi-monthly inspections has had a positive impact in the reduction ofcolonies with drone brood, thus reducing the potential for dispersal of unwanted genetics via males
that could mate with nearby colonies.

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The data revealed that when swarm traps were inspected within 2 to 3 weeks the number of combs
was less than swarm traps that had a longer interval between inspections. Table 1 below shows theresults of inspection for 3 traps captured at Honolulu airport this year and an average of the pastnumber of combs found in swarm traps when the inspections took place every month.
Interval Between
Inspection(days)

Number of combs(Brood -yes/no)

Oahu Trap 3

113 (NO)

Oahu Trap 12

21

4 (NO)

Oahu Trap 2

21

7 (YES)

Older Traps
Average (N=7)

30

6 (YES)
Range 4 to 7

Table 1 – Shows variation in comb production and brood development by swarms caught at the
Honolulu airport in relation to the time interval between inspections.
The images below show Swarm from Trap 12 (2022) (left) and compare it to a swarm from 2019(right) collected before the change in sampling schedule was implemented. The most importantfactors to consider are the number of combs produced and the percentage of the comb used in
reproduction – evident in the capped brood.

Trap # 2, 2022 - Notice the small size of 2 of thecombs
2019 Swarm Trap

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Sadly, a collection oversight or a holiday period that interferes with the inspection of a trap
can yield a larger population and more brood accumulating in a trap. Consequently, efforts
should be made to keep the 2-week interval.
The ideal situation is illustrated here– where the bees are established in the swarm trap andhave begun to construct comb but have not produced
large amounts of brood and especially no drones.
We would like to discuss the possibility of gathering
standardized data on the comb status (number of pieces),rough percentage of the comb with brood, as well, as an
estimate of bee population. The population can be easily
estimated by using jars with measuring lines that areequivalent to 100 bees or by using measuring scoops forcooking since it is already known that 1 cup is equal to
300 adult bees.

Swarm captures at airports
Swarm Captures per Island and Year
25
20
15
10
5
0
2017

2018

2019
2019
(Jan-May) (Jun-Dec)

Oahu

Maui

Kauai

2020

2021

2022

Big Island

Fig. 1 – Swarm capture rates/year and site. The year 2019 is divided in 2 since the protocol
for inspection frequency changed.

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Average swarm capture per inspection period
18
16
14
12
10
8
6
4
2
0
2017-20182020-2021
Oahu

Hilo

2022
Kona

Fig. 2 – Swarm captures/year by inspection period. During the 2017-2018 swarm trap
inspections were every month and the data for 2020-2021, as well as 2022, includes bimonthly swarm trap inspections. The year 2019 is omitted from this graph since it was theyear when inspection changed from once a month to twice a month.
Possible climatic correlates of swarm capture rates include total precipitation and seasonalityof rainfall. The rainfall can affect bees in 2 ways: 1- the bees have less time to forage andgrow slower, so feral and managed bees already established on island will
Rainfall in Hawaii is extremely variable depending on location. For example, on Oahu theaverage rainfall in the 2017-2018 period was close to normal after a previously dry year. In
contrast, in Hilo and Kahului the 2018 precipitation was well above average for the state.
Climate studies predict that rainfall levels in Oahu will decline by 30% or more in leeward
areas, however, the frequency of random weather events, such as cold fronts that can bringsudden rains, are expected to increase (Longman et al., 2001). Understanding the
population dynamics of feral colonies will require longitudinal capture data and weather
records. The information collected will provide valuable insight into swarming patterns at
each site. If Africanized bees were detected on island, this information, although based on
European bees, will most likely be applicable and will help anticipate swarming events
based on climate and related food abundance.
In addition to rainfall, plants and bees can be affected by ambient temperature. The apparent
decline in swarm capture rates on Oahu, Kona, and Hilo (Fig. 2) suggests a statewide
phenomenon. Rainfall in 2022 has been below average for most sites and temperatures in
Hawaii continue to rise. It’s possible that the low capture rate is related to the decline inabundance and/or health of feral colonies nearby.
5Obviously, the objective of the swarm traps is to attract a swarm arriving from outside
Hawaii, consequently the low capture rates of what could be local bees should not impact
the ongoing monitoring efforts. Invasion events are random, and the frequency ofoccurrence will not be tied to Hawaii’s seasonality. Nevertheless, it would be useful toevaluate the microenvironment in which these traps are located to make sure the dryweather and excessive insulation of the trap are not reducing our capture rates. If a trap isin full sun and the temperature within the trap is too high, this may reduce the appeal of thetrap to the scout bees looking for a potential nest site.
One recommendation, based on the current data, is to examine the location of each trap and if
possible, use a thermocouple to measure the temperature inside of the nest box andcompare it to the ambient at that point. A swarm trap should be a “nest site” that offers
shelter from the rain and excessive heat. It would be useful to invest in 2 thermocouples
with long leads so that the staff inspecting the traps can collect a reading “in-situ” for eachof the traps. Fluke thermocouples are the best for this kind of situation because they are
sturdy, portable, highly accurate, they can be adapted with a variety of temperature leads
and they give very fast readings (in Fahrenheit, Kelvin, and in Celsius) which is key forthis project. One of the thermocouples could be a travelling tool while the other one stays
on Oahu where most of the research is being conducted and we could create heating curves
for the traps under different shade cover conditions. However, for this to be successful theteam members on the other islands need to learn how to properly use the equipment toavoid getting false readings from direct sunlight exposure or wind effects on the lead,which is very sensitive.
Ensuring that the swarm lure is in good condition, that the trap is not getting excessively wetor too hot are crucial elements to the success of this project.

Bee Genetics and Invasive Subspecies
Import of live bees to Hawaii is illegal, and the only source of official new genetic bee material
imported to the islands is in the form of sperm brought by the queen breeders to produce specifictypes of bee races that the consumers on the mainland US are interested in buying from them.

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The Hawaiian archipelago is home to bees of European descent and no Africanized bee has been
detected in the past studies (Jara et al., 2014, and Szalanski et al., 2016) nor during the current
swarm genetic screening. We are currently focusing a large part of our effort to the screening for
Africanized genes in mitochondrial DNA. These tests provide us with information on the geneticsof the queen leading the swarm and/or the colony from which the worker bees come from.
The molecular test includes DNA
extraction followed by a process

Fig. 3 – Results from Restriction Enzyme on cytochrome

involving a restriction enzyme whichyields a “positive” or “negative” resultwith respect to Africanization. In Fig. 3 apositive control from an Africanized bee
from Arizona was used, and the local
samples show the expected 2 bars thatindicate “negative for Africanization”.
We know from past research that each island seems to have slightly different genetic profiles, allstill within the European Honeybee lineage. The subspecies that have been detected in Hawaii
include: 1-Lineage C, which includes bees of east European origin such as Apis mellifera ligusticaand Apis mellifera carnica; Lineage M, which includes bees of West Europe descent such as A. m.
mellifera and A. m. iberiensis.
Historically, Lineage C has been dominant (>80% of the bees sampled) on 2 islands: Oahu and the
Big Island of Hawaii belong to this lineage (Jara et al. 2014). This in part because of the great
concentration of queen breeders on Hawaii island and the past ease through which Oahu
beekeepers could obtain bees from these sources. In comparison in Maui, Kauai, and Molokai
about 50-60% of the bees belong to Lineage C and roughly 40% belongs to the Lineage M.
However, the access to queens from the Big Island of Hawaii by Oahu beekeepers has greatly
decreased. This is in part to new management in some of the queen breeding operations and lessinterest on their part to ship to small operations on Oahu. There also new regulations to between
island bee shipping that would require Hawaii queen breeders to have the boxes inspected for ants
before shipping to Oahu. This extra step in the shipping procedure and the relatively small ordersize from Oahu beekeepers makes selling to Oahu not attractive to these large companies and it has
become virtually impossible to get Big Island queens.
Consequently, we are working with the one registered, Department of Agriculture inspected, queen
rearing outfit on Oahu to examine their bees from a genetic standpoint. We have obtained DNA

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from 10 of their breeder colonies and these samples will be added to the ones from wild swarms(25 samples) and local private apiaries (25 samples). The DNA from these bees is now ready forsubmission to Eurofins, an extremely reputable company in the mainland that is used by other bee
researchers, for a more in-depth genetic analysis.
The results from those samples will allow us to examine if the genetics of bees in Hawaii havechanged in the past 10 years. Knowing what the allelic composition of the current honeybee
population is and how similar it is to the one we sampled 10 years ago will provide valuable
information if we were to detect anomalies in the genetics in the future.

The case of Jamaica
Most recently, I had the opportunity to assist the government of Jamaica in their efforts todetermine if they had Africanized bees now on the island. The data collected suggests that theAfricanized bees are now present in multiple areas in Jamaica and the government now is having toface making changes to the regulations that apply to commercial beekeepers and hobbyist urban
beekeepers. That experience has greatly influenced my perception of the threat and potential
impact of Africanized bees in Hawaii.

Development of Educational Materials
The program has developed a series of documents and video resources that await final review. Oneof the most difficult aspects is finding a suitable contact phone number – a line that will take after
hours and/or weekend calls as well. We also need to find a suitable website(s) for the materials.
There are also materials that await a few refining touches to become training modules forinspectors and extension agents. For a greater impact these materials need to be finalized with
input from the State Apiary Program and hopefully the HDoA.
As we expand into discussions with seaport authorities, we need more than ever to have a way toreach out to the workers, first responders, and the public in general about the threat of
Africanization of the honeybee population.

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