As a way of integrating the knowledge that we were gaining from the execution of the project with the knowledge of other people and acquiring their knowledge, we went after researchers, hospitals and the general population to compose this iGEM requirement.

During our research, we came across the production of the antiphid serum from the injection of snake venom in the horse. In view of this, a great doubt arose among us: what happens when the horse suffers an ophidian accident? To try to answer our doubt, we met with researcher and veterinarian Raquel Baccarin, who studies ophidic accidents in horses and the impacts that this brings to livestock.

At this meeting, we presented our project to the researcher and another guest, researcher José Paes, who is also researching cases of ophidian accidents in horses. During the meeting, the researchers presented to us cases of horses that were bitten by snakes and how the treatment was done. In addition, we were informed that these accidents are a big problem, because in many cases it is not possible to save the animal.

Figure 1 - Meeting

We learned that the antiophidic serum used for horses is lyophilized, which means that refrigeration is not necessary, and it can be used for any ophidic accident regardless of the species of snake. This happens because as the breeder only realizes that the animal was bitten by the snake after the symptoms appear there is no way to know which snake bit him.

We have also found that many of the accidents occur on the horse's snout, as the horse stays too long with its head down in order to graze. Accidents in this area are very harmful, because they make it difficult for the animal to breathe, which often causes death. Therefore, this meeting was of fundamental importance for us, as we better understood another side of the ophidian accidents, that of the injured animals.

Claudio Romeiro is a researcher and scientific influencer who promotes courses and lectures about snakebites and how to proceed and avoid these accidents. In addition, Claudio works at the Oswaldo Cruz Institute, the second largest producer of antivenom in Brazil.

Thinking about his great experience on the subject, we got in touch with him and set up a meeting to present our project and understand a little more about the subjects mentioned above. During the meeting, was discussed about large number of cases of snakebites and how the antiophidic serum its produce.

The researcher showed us how to proceed in case of snakebites and the production of antiophidic serum. At the meeting, the scientist also informed us how the characteristics of the snake's venom change as it grows, since the main purpose of the venom is to kill the prey so that the snake can feed.

In order to deepen our knowledge about the care of large animals, especially horses, we got in touch with Iris, a veterinary medicine student and volunteer at the Vale da Rainha sanctuary. This place is a recovery shelter for large animals that have suffered any kind of mistreatment.

Figure 2 - Meeting

The NGO is responsible for the medical treatment of physically exploited animals and also aims to prevent their euthanasia. As we have been informed by the NGO, the animals that suffer these abuses become useless to their owners and, consenquently, they end up losing their lives in the process. Thus, the NGO's goal is to rescue and treat the injured animals, as well as to offer them a free life.

In order to seek information about the treatment protocol for ophidic accidents and collect data such as: how often hospitals receive ophidic accidented victims, we visited some hospitals in Vale do Paraíba in the state of São Paulo. The hospitals visited were Santa Casa and Unimed, both in the city of Lorena.

During our research, we found that the incidence of these accidents is higher in the summer, probably because it is the most favorable time for outdoor activities. In Lorena and nearby towns, it is very common to hike in forests and this is where snakes come out to hunt.

We talked to the nurse Valéria, responsible for the infection control service at Santa Casa de Lorena since 2011, and we discovered that there is a protocol to be followed for the treatment of ophidic accidents. She told us that when the patient arrives at the hospital, a registration form, known as SINAN (Information System for Notification Worsening), is filled in to have an initial knowledge about the accident and the description of the animal. Then the patient's symptoms and their evolution over the days are evaluated. Fever, body aches and increased heart pressure are the most frequently seen mild symptoms.

The nurse also told us that in cases of accidents with venomous animals, treatment with serum therapy should be started as soon as possible, after a brief assessment of the type of snake, for a more precise indication of the correct serum to be used. Because of this, the hospital's nurses have 24-hour contact with biologists from the Butantan Institute that can identify the snake based on its bite and, consequently, help in the treatment.

According to her, this therapy is a treatment made from a serum and should be started within a maximum of 2 hours after the snake bite. In the first 30 minutes, corticosteroids are injected into the patient, and then the serum ampoules are diluted in a proportion of 10 ml of antivenom (pentavalent antibothropic serum, in the case of Bothrops jararaca) to 20 mL of glucose solution or saline. In the hospital, glucose serum is more commonly used, because it allows a faster effect on the body. The average observation time of patients affected by ophidic accidentes by non-venomous animals is 6 hours; however, for cases with venomous animals, the minimum observation time is 12 hours. The patient remains in observation state until released; Blood and other materials such as urine, for example, are constantly collected to monitor the kinetics of the poison in the body. If the patient does not show improvement and his symptoms continue to evolve and become more severe, the possibility of admission to the ICU is considered.

Futhermore, we had access to the treatment records of patients who had suffered from snakebite at Santa Casa in the last six months and we could see that serum therapy was used in most cases. As a way of complementing the results obtained with the hospital data, we created a forms about snakebites on our social networks and WhatsApp groups in order to seek data not only from Lorena and region, but from various parts of Brazil, increasing thus our basis for better conclusions about accidents in the country.

In order to take our project to as many places as possible, we presented Honorato in several schools and in several different places. Honorato was in 2 states of Brazil and in 3 different cities, taking knowledge about synthetic biology and making people aware of ophidic accidents. This is the distance from our college to all the educational institutions where we have given presentations:

As a way to publicize our project, we made a presentation at the Cooperpro Objetivo school in the city of Casa Branca – SP. We present the first class of the synthetic biology tournament to high school students.

Figure 3 - Presentation at School

Students also participated and engaged the conversation mainly in the part of snakebites in animals. We had several reports of snakebites in dogs and horses.

In conclusion, the presentation was terrific, we were able to present about the knowledge regarding synthetic biology and also explain our project, but the experience was reciprocal, since we also received knowledge from the students' experiences.

As a way of spreading knowledge about science and awareness about snakes, we went to Colégio Nossa Senhora de Fátima, in Vitória da Conquista – BA. The presentation was essentially for high school classes and we brought applied biology and genetics as a guide, and with the applications and demonstration of what we can do with these techniques, there are always doubts and questions from students.

In addition, again, we compared common sense knowledge, coming from students, and scientific knowledge. With this, we were able to demystify several misconceptions about snake and snake accidents, in addition to being able to pass on correct first aid practices in cases of ophidic accidents.

Figure 4 - Presentation at School

Invited by the professor of Biochemical Reactors discipline for the undergraduate environmental engineering course, we went to make a presentation as a way of exemplifying concepts based in the classroom and the scope of our project at the Federal Institute of Bahia, in Vitória da Conquista - BA.

The students, in addition to being delighted with our project and what synthetic biology allows us to do, engaged mainly in the part where we talked about snakebites, adding their knowledge to the conversation.

Together with "Associação Marie Curie Vestibulares", our team USP-EEL-Brazil went to present our project at Arnolfo Azevedo school, during the joint project between the school and the student organization "The power of engineering". In this presentation we talked about ophidic accidents and how we should proceed in such cases. We also did a quiz with the children after the presentation and those who got the right answer got a candy.

Figure 5 - Presentation at School

The participation of the students was of fundamental importance for the presentation. At the beginning of the presentation we asked if anyone have already had a snake accident in their family or in people they knew. One of the children answered positively and that his grandfather had been bitten and died. Even though everyone on the team was a little scared, we were able to show how serious ophidic accidents are and how important it is to follow the recommendations we presented to them.

The talk with the students was very satisfying, since they shared their experience with us and we transmitted information to them. Also, during this visit, we saw the drawings that the children had made and in them there were some snake drawings, which made us very happy, since we saw our project being remembered. At the end of the engineering project, we were invited to present again another time.

Figure 6 - Snake Drawing

In order to better understand the ophidic accident scenario, both from a national perspective and beyond our borders, we set up a form to collect reports related to these accidents. In addition to sharing it via our university's institutional email, aiming to reach the entire academic community, we also sent it to the iGEM IISER-Pune-India team and to the Chihuahua City team in Mexico.

We have received several personal experiences regarding snake bites and their care and treatment. It was reported to us that a person was bitten by a snake in a relative's backyard during a family lunch. When they went to the nearest hospital, there were no conditions necessary for treatment, and no ophidic serum was available. Thus, the person was taken to another hospital in the neighboring city and there received the serum and pain medicine. However, the next day the swelling and pain remained and together with the parents, the person went to a third hospital for medical consultation. There it was noticed that one toe was necrosed and they continued the treatment. Amputation was not necessary. The same person who was stung also reported that his aunt had suffered an ophidian accident years before.

Another critical account that we received from an Indian person was that a friend, in December 2021, was bitten by a snake and was taken to the hospital, where they noticed that the case was serious. They started treatment, and 2 to 3 toes were amputated, and after the person was kept under observation in the hospital, they died. This case was in a Village near to Haveri (Karnataka, India).

We have received several reports from relatives of people who have been bitten on farms, or in vacant lots, or even in backyards where there is a forest nearby. Most of the victims managed to find a hospital that offered the ophidian serum, but there were cases where sequels were observed. For example, purple marks near the site of the bite, pain in the days following the accident, or even amputations. One report we collected was of a man who, after being bitten, ran back home to inform his family and be taken to the hospital, but this caused the venom to spread more quickly in the blood and consequently his leg was amputated.

We also collected experiences about how lack of information and instruction can lead to more serious sequelae when it comes to ophidian accidents. In one of the cases, a tourniquet was performed at the site of the bite, causing the poison to concentrate there. Because of this procedure, done before arriving at the hospital, they lost part of their finger.

With the answers we received on the form, we can better analyze the context of ophidian accidents and understand how people react, what the consequences are, the most common sequelae and also treatment taken in hospitals. This data collection has greatly helped widen the team's vision and encouraged us to promote more informative and instructive posts and plan conversations with schools, farmers, nurses, victims etc. in order to get the information out to as many people as possible.