Pollination and Paternity Testing

By Holly Brabazon

Dr. DeWayne Shoemaker and Holly Brabazon of University of Tennessee, Knoxville


Would you believe that small biting midges are cacao’s main pollinator? They’re only about the size of a pin head, and they don’t fly very well either, but with how small cacao flowers are, and how they’re shaped, cacao has to rely on the tiny midge to pollinate its flowers. Unfortunately, midges worldwide do a poor job pollinating all the flowers made by cacao trees. Only a small percentage of flowers get pollinated, and even then, only a few of those pollinated flowers receive enough pollen (about 115 pollen grains) to make a fully formed, viable pod.

A first step to better understand pollination in cacao is to study pollination of wild-growing cacao. To date, only a few studies exist on wild cacao pollination, and many questions about pollination remain unanswered. For instance, we still do not know how many times a flower needs to be visited to be fully pollinated, how often flowers are pollinated with pollen from the same tree, or how pollen is moved around the forest. Not to mention, we only have a rough guess of how far a pollinator can even travel. These are some of the questions I will investigate as part of my PhD research.

To start answering these questions, we will gather genetic information from all the wild cacao trees growing in the forest at BFREE by sequencing the DNA of these trees. There are about 300 wild trees at BFREE, and thanks to Elmer Tzalam, Mark Canti, and the cacao team’s hard work, each and every tree has been visited, georeferenced using GPS, and labeled with a metal tag. We were able to use these GPS coordinates to find and collect leaves from many of the trees at BFREE in June 2022. Now, Lenardo Ash, a BFREE Cacao Science Fellow graduate and student at the University of Belize, has taken on the responsibility of collecting leaves from the remaining trees. Once the leaves are collected, we will extract and sequence their DNA. These genetic data will allow us to identify unique DNA tags for each tree, like a fingerprint, from the unique patterns in their DNA.

With these DNA “fingerprints,” we can learn a lot about the natural history of the cacao growing in the forest at BFREE. We will see how genetically diverse the population is and determine if there are unique clusters of trees with similar genetic variation. Many other cacao populations in the world have genetic mechanisms to prevent self-pollination, and we will see if those same genes are found in the BFREE trees.

We also will use these genetic data to figure out exactly which trees are pollinating other trees’ flowers, just like a paternity test. To perform our paternity experiment, we collected pods from several wild cacao trees growing out in the rainforest at BFREE. We brought those pods back to the nursery and planted the individual beans in bags of soil, making sure to carefully label each soil bag to indicate which tree the pod came from. The beans are growing in the nursery right now. Once the seedlings have leaves big enough to collect, we’ll sequence their DNA. Then we’ll do a paternity test on each seedling to identify who the father tree is that contributed the pollen. Once we know who the father tree is, we’ll map how far pollen traveled to pollinate the flower on the mother tree. This information will allow us to search for patterns of pollen moving around the forest. With pollination being a limiting factor in cacao production, our studies may ultimately help cacao farmers increase production with better pollination management. We just need to better understand how cacao pollinators move around the forest and what they are capable of in a natural rainforest environment.

We had an amazing time working with all the great people at BFREE. It was an adventure to explore a pristine rainforest searching for cacao trees, and I can’t wait to see what we learn from our results!

Cacao Fellow, Mark Canti, Explains the Process of Adopting a Tree from the BFREE Farm:

By Mark Canti

Hello, my name is Mark Canti. I’m the BFREE Cacao Fellow, and I oversee the cacao adoption program at BFREE in collaboration with the Heirloom Cacao Preservation Fund. I’m always very excited when I learn that a new tree has been adopted from our farm, and I am eager to tag the newly adopted tree. 

I first create a personalized tag for the tree by engraving the adopter’s name or the adopter’s chosen honoree on an aluminum tag. Then I grab my gear, including the newly created tag, a GPS device, and my camera. Next, I need to select the perfect tree. I’m looking for healthy trees that have at least 70% shade and are at least 1-1.5 meters tall. Once the tree has been selected, carefully tie the tag to a tree branch and record the GPS coordinates. Finally, comes my favorite part of the process. I’m very passionate about photography, and I really enjoy the opportunity to photograph each tree. My dream is to capture wildlife such as a beautiful bird like a warbler when I’m taking each photo. I like that the pictures I take can help the new adopters feel as close to being on our beautiful farm as possible. 

I’m very proud to be part of the Adopt a Tree program, and I would like to thank everyone who has adopted a tree from our farm so far. I hope I have the opportunity to select and photograph a tree for you! 

If you would like to adopt a tree from the BFREE Farm, please visit the Heirloom Cacao Preservation Fund’s website and select HCP#11.

Adopt a Cacao Tree – HEIRLOOM CACAO PRESERVATION FUND (hcpcacao.org)

Celebrating Earth Day

Students from Keene High School in Keene, New Hampshire helped BFREE staff celebrate Earth Day by planting seeds. This is Keene High School teacher, Matt Brady’s, fourth trip to Belize and to BFREE. He is joined by fellow teachers, Christine Gillis, Monica Foley, and Jodie Ballaro. Their group was scheduled to come to the field station in 2020 but was cancelled due to the pandemic. They tried again last year with no luck. This makes us especially thrilled to host them in 2022.

In a BFREE interview with Mark Canti and Jonathan Dubon on Facebook Live for Earth Day, Matt described why he wanted to return. “BFREE is a really special place for lots of reasons. I’m really happy to be here to meet young people like you. People who contribute to the ecology of the area and are conservationists. That is very important to me, the way BFREE is set up to keep young people coming in from the area. This is why we keep coming back.”

In a surprising turn of events for dry season, it began raining at 9am during the student orientation. The rain continued throughout the morning and into the afternoon, but this didn’t dampen anyone’s spirits. The students continued their orientation and tour of the facilities. After lunch, everyone divided into groups for service projects. Nine students helped with planting germinated cacao seeds in the nursery. An additional fourteen students helped at the Hicatee Conservation and Research Center where they assisted in a project to improve the exterior fence. The remainder of the students supported the long-term large mammal research project by checking camera traps on the property.

We are grateful that Matt, Christine, Monica and Jodie worked so hard to come back to BFREE this year.

Student Groups Return to BFREE After Two-Years

It is no secret that the global pandemic has significantly impacted travel. For BFREE, that has meant a drastic reduction in all types of visitation over the last two years from researchers, specialists, eco-tourists, to our annual Field Courses. However, things are slowly but surely changing, and we are ecstatic to have had our very first student groups back at BFREE since February 2020!

Nearly two years to the day since our last student group, we welcomed two groups from an organization new to BFREE, ARCC Gap Year Abroad. ARCC offers summer and gap year programs for students focused on community-driven and community-led sustainable projects around the world. The two student groups visiting BFREE alternated between 5-nights at BFREE and 5-nights on the Belize Barrier Reef. Each group was made up of ten students and two instructors. The groups volunteered with BFREE’s cacao agroforestry program supporting the growing nursery and painting signs for trees on the farm. Additionally, students learned to make chocolate from bean to bar.

Other highlights include being the inaugural guests to sleep in the newest accommodations at BFREE, The Hammock, exploring the Rainforest on a boundary line hike with Protected Areas Manager, Canti, and meeting and learning from all of the incredible and inspiring staff at BFREE.

These students are part of a 70-day program that will take them from the Rainforest to the Barrier Reef, to sea turtle restoration and surfing in Costa Rica, and finally exploring the Panama Canal over the next two months.

What an amazing adventure these students are on – wishing them all a safe and incredible trip!

Below are a collection of photos taken by BFREE staff and volunteers of both the ARCC GAP GROUPS A and B during their time at BFREE in February 2022.

Autumn Dietrich Completes 10-week Internship at BFREE

BFREE internship opportunities have returned since the start of the COVID-19 pandemic. After more than two years of not hosting interns, Autumn Dietrich is welcomed with open arms to BFREE for a 10-week program from January to March 2022.

Autumn is no stranger to BFREE. She first visited on a Field Course as an undergrad with Western Michigan University in 2018. On a hike with Protected Areas Manager, Sipriano Canti, Autumn describes the experience as having changed her life. Impressed by the sheer scale of life that existed at BFREE, she hiked with her fellow classmates winding through trees, climbing embankments of creeks, and cutting through giant leaves. The hike led the group to the Ranger Observation Post on the boundary of BFREE and the neighboring farming village of Trio. Autumn’s excitement turned to dismay as she witnessed for the first time the harsh boundary line between a healthy and thriving forest and a large expanse of farmland. At this moment, she felt an overwhelming feeling that she wanted to help make a difference and help support protected areas. She knew she would come back to BFREE.

After graduation, Autumn applied to return to BFREE as an intern for three months, April – June 2020. At the time, we had no idea we would soon be facing a global pandemic. Determined not to give up on her dream to return to BFREE, Autumn’s postponed trip came to fruition this year when she traveled to Belize in January.

Autumn worked alongside BFREE staff during her ten-week internship, providing invaluable support. Her first week was spent documenting and assisting a research crew from Kutztown University. They are establishing a long-term experiment that quantifies the influence of Yucatán black howler monkeys on biodiversity and ecosystem function in Belize. She then assisted Cacao Fellow, Mark Canti on a large-scale survey of the cacao farm to determine the farm’s overall health. When not focused on collating the cacao survey data, Autumn assisted Housekeeping Manager, Ofelia Cus with preparing accommodations for visitors and BFREE Chef, Edwardo Pop in the kitchen.

In addition to supporting BFREE’s various conservation initiatives, Autumn, immersed herself in learning and experiencing Belizean culture. She was honored when Operations Manager, Elmer Tzalam and his wife Gina invited her to spend a weekend with them at their home in Golden Stream. Autumn loved spending time with Elmer and Gina’s two children, Esther (12) and Travis (9), and enjoyed a delicious homemade dinner of panades, a dish that’s similar to empanadas but with a Belizean and Mayan twist.

Living full-time at BFREE for ten weeks brings plenty of challenges. The remote location and intense heat are not for the faint of heart. However, Autumn not only thrived during her two+ months, but she also brought so much joy and laughter to the BFREE community. Thank you so much, Autumn, for your sincere dedication to BFREE’s mission and for being an incredible and loyal team player. We all look forward to seeing you amongst the giant ceibas again one day! 

Meet BFREE’s Newest Fellow, Mark Canti

Mark Canti in BFREE’s Cacao Nursery, August 2021

A Little About Me
I was born in a subtropical climate during the rainy season in my native village of Golden Stream. It is located along the main highway, a couple of miles south of the BFREE junction. I do not remember much of my childhood, but I sure remembered how much my parents loved, cared, and supported me throughout my childhood. I attended primary school graduating as a salutatorian. I moved on to high school with a mindset of “Oh, I’m just gonna do whatever.” I wasn’t involved in anything. After graduating from high school, I wasn’t planning on going to college, so I stayed home doing chores and other temporary jobs such as construction, woodwork, and maintenance. Over that period of time, I attended summer camps with Ya’axche Conservation Trust, whereby I first started to develop a sense of interest in nature.

The following year I applied to Independence Junior College, majoring in Natural Resource Management (NRM). It took me a little while to commit myself to education but, once I did, I was invested despite the lack of internet access and technology at home. While attending college, I became extremely involved on campus by volunteering to plant trees and attending clean-up campaigns with non-governmental organizations like the Toledo Institute for Development and Environment (TIDE) and Oceana.

Most of my favorite hobbies are related to the life of an environmentalist. I like nature walks, night hikes, mountain climbing, canoeing, traveling, photography, and snorkeling. Soccer is my favorite sport simply because it helps me stay active, allows me to socialize, and for the most part, it assists me in clearing out stress. I’m an easy-going individual who is focused on conserving the environment and developing advanced photography skills.

First Memory of BFREE
My first memory of BFREE was one and a half decades ago when I was a kid attending primary school. I remembered coming to BFREE on an educational school trip where I first witnessed Mr. Jacob Marlin display an amazing activity where he captured a venomous snake called a fer-de-lance. He was so generous that he gave us the experience of touching the snake, which is impossible for a child to do on its own. That was one of the greatest experiences I have had as a child.

Another Visit to BFREE during College
During my years of study at Independence Junior College, I never imagined I would be working for BFREE one day. Not that I wasn’t interested, of course, I always thought about it during the final days of the last semester at IJC. What really motivated me was attending a school trip here at BFREE, whereby a sensational occurrence happened. Guess what? We were the first set of IJC students to get the opportunity to see the Harpy Eagle with our sharp, naked eyes. So that wonderful experience made me curious and more interested in wanting to work and join the BFREE family to help support mother nature.

Over the Next Two Tears
One of the things I have noticed about BFREE is that it has been providing opportunities for the Fellows to improve their writing skills by allowing them to participate in helping write reports and grants. This is interesting to me. I am also interested in working with researchers, which allows the Fellows to meet new people while also learning advanced research assessment skills, which could be useful both for the organization and a Fellow’s own career.

What I like About Cacao-Agroforestry
With cacao-agroforestry, I’m most interested in how interplanting cacao trees, along with shade trees that bear fruit and other hardwood trees, attracts different species of birds and other animals. Programs like this, which seek to regenerate the rainforest, are both beneficial to our well-being and also to the environment. Healthy forests provide us with cleaner air and also, ultimately, prevent animal species from going extinct. Separately, the program also allowed me to unlock a skill of mine I never knew I had, which is grafting cacao trees.

First BFREE Cacao Fellow Completes Program!

BFREE Staff celebrate Lenardo’s last day as the Cacao Fellow on Thursday, August 19th.

BFREE’s first Cacao Fellow, Lenardo “Leo” Ash, is graduating from his two-year work-training program this week. He will immediately begin studies at the University of Belize, where he will work toward his Bachelor of Science in Natural Resource Management. Lenardo began his Fellowship in July 2019 under the mentorship of Cacao Program Director, Erick Ac. He spent the remainder of that year “learning by doing” and was completely immersed in all things cacao agroforestry. By early 2020, Leo was well-versed enough on the topic that he was able to start co-presenting to BFREE’s visiting students and researchers.

In March 2020, the COVID-pandemic closed BFREE to visitors and a by-product was the uncertainty of the continued employment of all of BFREE staff. When land borders closed, Erick Ac was no longer able to travel from Guatemala to Belize to oversee the cacao program. Unfortunately, the academic components of Leo’s program fell to the wayside for a while as BFREE’s administrative staff focused energy on ensuring the safety of the BFREE staff and finding the financial means to keep as many people employed as possible.

In spite of the lost opportunities for his professional development including canceled travel plans, research projects, and conferences, Lenardo showed great determination in maintaining his path toward personal and professional growth. Lenardo began practicing Spanish during virtual weekly meetings with Erick, he birded with other BFREE staff, and he participated in Herpetology 101 learning the Scientific names of all the turtle and lizard species on the reserve. He asked for reading assignments to expand his knowledge on cacao and agroforestry and eagerly accepted any opportunities to give virtual presentations to BFREE audiences.

Because of his strong interest in photography, Lenardo began photographing birds and other wildlife around the property. Last July, he spotted a ten-foot boa constrictor attacking a turkey vulture and immediately ran to get a camera and to notify other staff. Images and videos that Leo took of the predation event helped provide details for a scientific article, which will be published in the September 2021 issue of the Journal of Raptor Research.

Earlier this year, Lenardo was invited to be a part of a research team hired by the Heirloom Cacao Preservation Fund. Along with researchers from all over the world, Lenardo spent nearly six months compiling a literature review that explored cacao genetics across the globe.

Lenardo has never given up his dream of continuing his education beyond his Associate’s Degree, so he applied to the University of Belize and was accepted for August 2021 admission. Although, we are sad to lose such a valuable team member, we are excited about Lenardo’s bright future and can’t wait to see where his journey will take him.

Platt, S.G., Barrett, H.A., Ash, L., Marlin, J.A., Boylan, S.M. and Rainwater, T.R. Predation on Turkey Vultures (Cathartes aura): A New Observation and Review, Journal of Raptor Research. Vol. 55(3), September 2021. Pp. TBD

Congratulations to Jaren Serano, BFREE’s first Wildlife Fellow alum!

Congratulations to BFREE’s first Wildlife Fellow alum, Jaren Serano, who recently graduated with honors from Jacksonville University. He completed his Bachelor of Science in Sustainability and Minor in Biology in June 2021.


Jaren helped launch the BFREE Science and Education Fellowship Program in January 2018. With the support of the Turtle Survival Alliance (TSA), Jaren spent the two-years in the new work-training program. He learned to support the operations of the HCRC and he also had the opportunity to glean knowledge from the many amazing visitors to the field station. He participated in and presented to field courses with students from all over the world, he assisted visiting researchers and helped implement outreach programs.


During his second year, he began presenting at professional conferences in Belize. In August 2019, he traveled to Tucson, Arizona with Tom Pop, Jacob Marlin, and Heather Barrett to present at the 17th Annual Symposium on the Conservation Biology of Tortoises and Freshwater Turtles. Jaren’s ten-minute talk received a standing ovation and he was awarded Best Student Presentation. That symposium was critical to Jaren’s next steps because the team re-connected with Dr. John Enz of Jacksonville University (JU) who brings student groups to BFREE. John learned that Jaren was applying to schools in the U.S. to complete his Bachelor’s degree and suggested that Jaren apply to JU.

With the help of an amazing GoFundMe campaign, which many of you supported, and a substantial scholarship from JU, Jaren was able to enter college in January 2020 – just in time for a global pandemic. In spite of many challenges, Jaren excelled in his courses and was an active contributor in the classroom and a role model to other students. During his summers, Jaren returned to Belize and BFREE where he assisted with field research relating to the Hicatee, helped with projects at the HCRC, and, most recently, participated in the TSA-North American Freshwater Turtle Research Group (NAFTRG) turtle survey of the BFREE reserve.

Jaren will begin graduate school at the University of Florida this month. Jaren received a full-tuition scholarship through the Doris Duke Conservation Scholars program and secured a research assistantship to cover additional costs. Jaren will work under the advisement of another BFREE partner, Dr. Ray Carthy, in the department of Wildlife Ecology and Conservation. Jaren’s research will examine how human interventions such as beach renourishment impacts natural coastal processes and resilience. Primarily, he will examine how gas exchange relates to sea turtle nesting, dune building, and carbon sequestration.

We are incredibly grateful to all those who have supported Jaren and cheered him on throughout his journey. A special thanks to Turtle Survival Alliance’s Board of Directors without whom Jaren’s Fellowship would not have been possible. Also, to John Enz and Ray Carthy for being incredible BFREE partners and mentors to the BFREE staff. Thanks to Day Ligon and Denise Thompson for their support and tutelage of Jaren and other BFREE staff over the past few years. Finally, thanks to the many donors who supported Jaren’s GoFundMe campaign. Each and every one of your gifts mattered!

Wildlife Fellow, Jonathan Dubon Embarks on 2nd Year

Jonathan Dubon is the second Wildlife Fellow to take part in the BFREE Fellowship Program. Jonathan began working with BFREE in June 2020, immediately after the shelter-in-place order was lifted in Belize. He assists Tom Pop at the Hicatee Conservation and Research Center (HCRC), and although his program began during the COVID pandemic, his first year has been a productive one.

Under the mentorship of Tom Pop, Jonathan has learned how to feed and care for all turtles at the HCRC. He has learned to look for signs of stress and illness and to collect morphometric data during the bi-annual health assessments. He has worked closely with Tom on several projects to upgrade the facility; the most notable have been improvements to the nesting areas and the water movement.

Additionally, Jonathan has been responsible for updating and managing an ongoing census of the captive population of Hicatee turtles in residence at the HCRC. With hundreds of turtles hatching each year and with the recent turtle releases, this is no easy task. The information is constantly changing, so he has to review the data regularly to ensure accuracy. He has taken on the essential responsibilities of creating quarterly reports of activities affiliated with the HCRC, water quality testing at the HCRC, and managing the weather data for the entire field station.

Working with radio-tracking devices to better understand what happens to hicatee in the wild.

Over the past year, Jonathan has both led and co-presented on several virtual presentations. On August 27th at 12:15 EDT, he will present virtually at the 19th Annual Symposium on the Conservation of Tortoises and Freshwater Turtles. His talk is “Environmental Education and Re-wilding of the Critically Endangered Central American River Turtle (Dermatemys mawii) During the 2020 Pandemic.” He will discuss his role in the newly formed Hicatee Awareness Month planning committee and will highlight last October’s awareness campaign. He will also describe the release of the HCRC’s first re-wilded Hicatee turtles and one of the associated community outreach events. To learn more about attending the conference or to see Jonathan’s presentation visit our partner’s website and register for free here:

https://turtlesurvival.org/2021-symposium/

Jonathan has spent the last month in the field monitoring the movements of 25 recently re-wilded turtles from the 2021 hatchling cohort. He and our partners from Belize Turtle Ecology Lab are radio-tracking turtles to begin to understand how far they travel from their release point, what their habitat preferences are, and how they fare after being released.

In addition to turtles, Jonathan has an interest in snakes and large cats. In his final year as Wildlife Fellow, he hopes to work more directly with the large mammal camera trapping project. With new Panthera cameras stationed throughout the property, his involvement will likely become a reality in the very near future.

Introducing Jonathan Dubon, BFREE Science & Education Fellow

Jonathan Dubon, BFREE Science & Education Fellow

BFREE is pleased to introduce our newest Science & Education Fellow, Jonathan Dubon. Jonathan grew up in Independence Village about 20 miles east of BFREE and has known from an early age that he wanted a career that would include his passions for field experience and outdoor adventures. This passion grows from visiting his Grandma’s farm near Punta Gorda as a child where he has many fond memories of exploring her land and being exposed to nature. Because of this childhood experience and influence from his brother who is also involved in conservation, Jonathan went on to study Natural Resource Management at Independence Junior College. He graduated with his Associate’s Degree in June 2019 and with the highest honors in his department.

Jonathan’s first visit to BFREE was on a school field trip with Independence Junior College in February 2019. Jonathan says, “I fell in love with the place and it’s environment – at that very moment I knew I wanted to come back. I like everything about being at BFREE including the friendly staff, the environment, everything is just very welcoming. This is exactly where I imagine my dream job.” He returned one year later as a volunteer in the Spring 2020 Hicatee Health Assessments where he assisted in the 5-day health check.

Jonathan, second from the left, back row, along with fellow classmates from IJC on a field course at BFREE in February 2019.

Now, in the second week of his fellowship, Jonathan shares, “It’s so exciting to be here at BFREE right now. I only know a little bit about the biology of Hicatee Turtles and I am overly excited that every day I now get to learn something new about them. It is thrilling to work with the hatchlings; I am also eager to learn about all the other animals found here at BFREE such as birds, snakes, and mammals. I also really enjoy hearing the birds singing early in the morning while working by the pond. “

Jonathan says, “my message to all Belizeans is that the Hicatee are especially important to our ecosystem, and it is critical that we protect them – Belize has the honor of being the final stronghold for these turtles, who are the last in their lineage. “

We are thankful to the Turtle Survival Alliance for their funding of the BFREE Science and Education Fellowship. This is the second fellowship funded by the TSA; the first was awarded to Jaren Serano who served as the BFREE Science and Education Fellow from January 2018 – December 2019. The Science and Education Fellowship is assigned to support the operations in one of three areas at BFREE – the Hicatee Conservation & Research Center, the cacao agroforestry project or the protected areas program. It is a two-year immersive work training opportunity for recent Belizean junior college and college graduates who exhibit leadership potential combined with a clear interest in the conservation of the country’s natural resources

Jonathan hands an adult hicatee turtle from the breeding pond to BFREE Executive Director, Jacob Marlin during the Spring 2020 Hicatee Health Assessments.