Biosecurity practices and sanitary challenges in dairy farms in BrazilABSTRACT:
The research applied an online questionnaire about farm characteristics, sanitary and manure management, carcass disposal, reproductive and milking practices, and farmers’ perceptions of disease prevention. Farms were classified by milk production and production system. Less than half of the respondents (41.8%) had previously participated in any training on biosecurity or disease prevention, and most (94.5%) consider prevention to be more advantageous and less time-consuming than disease treatment. Small and medium-sized farms had a lower level of biosecurity practice utilization compared to larger farms; although, critical practices are followed on these farms. Preventive measures, such as performing diagnostic tests (P < 0.05) and quarantining newly acquired animals (P < 0.05), were significant in larger herds and intensive production systems, respectively. Practices related to female and newborn management, such as maternity pasture (P < 0.05), were described in intensive systems. There were associations between large and intensive farms performing colostrum feeding only with a colostrum bank (30%), and extensive farms with lower milk production using the natural colostrum method (70%). The collection and storage of manure are carried out by most farms (74%), although only 9 (10%) subject it to any type of treatment. The data suggested that, in larger farms and intensive systems, more preventive measures and actions related to herd health are carried out. Given the variety in the application of these measures, awareness, and technical assistance are necessary to conduct and apply specific biosecurity measures according to the resources and needs of each farm.
Key words:
biosecurity; dairy cattle farming; production systems
RESUMO:
A pesquisa aplicou um questionário online sobre as características das propriedades, manejo sanitário e de esterco, descarte de carcaças, práticas reprodutivas e de ordenha, e percepções dos produtores sobre prevenção de doenças. As propriedades foram classificadas por produção de leite e sistema de produção. Menos da metade dos respondentes (41,8%) havia participado anteriormente de qualquer treinamento sobre biossegurança ou prevenção de doenças, e a maioria (94,5%) considera a prevenção mais vantajosa e menos consumidora de tempo do que o tratamento de doenças. Propriedades de pequeno e médio porte utilizam práticas de biossegurança em menor nível em comparação com as de maior porte, embora práticas críticas sejam seguidas nessas propriedades. Medidas preventivas, como a realização de testes diagnósticos (P < 0,05) e o isolamento de animais recém-adquiridos (P < 0,05), foram significativas em rebanhos maiores e sistemas de produção intensivos, respectivamente. Práticas relacionadas ao manejo de fêmeas e recém-nascidos, como o pasto de maternidade (P < 0,05), foram descritas em sistemas intensivos. Foram encontradas associações entre grandes propriedades e sistemas intensivos que realizam a alimentação com colostro apenas com banco de colostro (30%), e propriedades extensivas com menor produção de leite que utilizam o método natural de colostro (70%). A coleta e armazenamento de esterco são realizadas pela maioria das propriedades (74%), embora apenas nove (10%) o submetam a qualquer tipo de tratamento. Os dados sugerem que, em propriedades maiores e sistemas intensivos, são realizadas mais medidas preventivas e ações relacionadas à saúde do rebanho. Dada a variedade na aplicação dessas medidas, é necessário conscientização e assistência técnica para a implementação e aplicação de medidas específicas de biossegurança de acordo com os recursos e as necessidades de cada propriedade.
Palavras-chave:
biosseguridade; bovinocultura de leite; sistemas de produção
INTRODUCTION
Dairy cattle farming and the production of milk and dairy products hold great economic and social importance for the state of Minas Gerais and the country. Brazil stands out as the 5th largest milk producer in the world and has great potential as a dairy exporter due to its climatic, market, and geographic conditions, among other characteristics (FAO, 2023). Preventing the introduction, development, and spread of transmissible diseases in the herd, primarily through the adoption of biosecurity practices, is necessary to ensure greater profitability for the production system, guarantee animal health, preserve public health, and ensure food safety.
The health and well-being of cows and heifers determine the profitability of a dairy farm, as only a healthy herd can reach its maximum productive potential and perform effectively. The incorporation of biosafety policies based on risk assessment and management into the herd health management program helps reduce the introduction and spread of infectious diseases within farms (BUSHRA et al., 2024).
Biosecurity in the context of animal production can be defined as a set of technical measures aimed preventing the introduction, establishment, and spread of diseases, infections, or infestations within an animal population (LIMA-CAMPÊLO et al., 2024). In this regard, it includes both external and internal measures, related to actions designed to prevent the entry or exit of pathogens. Therefore, all these measures are intended to reduce the spread of these pathogenic agents within the herd (RENAULT et al., 2018). Established biosecurity practices on dairy farms ensure healthier and more resistant animals, improve animal welfare, reduce the spread of zoonotic diseases, and decrease the use of medications, such as antibiotics; consequently, improving the quality and safety of the final product (PEGORARO, 2019).
In Brazil, studies on the implementation of biosecurity in cattle farms are rare, which is noteworthy given the importance of the topic of milk production, the diversity of systems, and the need to conduct and apply specific measures for each farm to ensure a safe and high-quality final product, preserving animal health and strengthening One Health (FRANCO et al., 2021). Moreover, a strategic biosecurity program needs to be more than just a list of generic tasks; it is imperative that it be decision-focused and flexible enough to adapt to the unique situations of individual farms (PEGORARO et al., 2018).
Due to the importance of biosecurity in ensuring animal health and milk production quality, as well as the scarcity of studies on its implementation in Brazilian dairy farms, this study analyzed the adoption of biosecurity practices in dairy farming. Using an online questionnaire, it investigated the measures implemented and their relationship with variables such as herd size and farming system, contributing to the development of more effective and adaptable biosecurity strategies.
MATERIALS AND METHODS
The research consisted of administering an online questionnaire, followed by a descriptive and analytical statistical analysis of the results, and a discussion of the main findings to understand which measures are adopted and the main reasons related to the presence or absence of biosecurity in dairy farming, as well as its relationship with variables such as herd size and type of farming system.
The data regarding biosecurity practices on dairy farms were collected through a questionnaire developed using the Google Forms application and approved by the Research Ethics Committee (CEP) of the National Health Council (CAEE: 57307822.1.0000.8158). After approval, the questionnaire was distributed via social media and private groups between September and November 2022, targeting rural producers, owners, or those responsible for the dairy operation of the farm.
The questionnaire used in this study was developed based on a previously validated model by RENAULT et al. (2018) and DAMIAANS (2019), which originally contained 304 questions, and was adapted to consist of a total of 54 questions. The questionnaire considered information about the farm and its respective representatives; sanitary management and protective measures of the farm; manure management; disposal of dead animal carcasses and veterinary waste; reproductive and milking management; and farmers’ perceptions of disease prevention and control in the herd. Furthermore, for better data coding and analysis, the participating farms were classified according to herd size based on milk production and type of farming system.
Based on the extensive practical experience of the research team in Brazilian dairy cattle production, the questionnaire was adapted to focus on reducing the number of questions to facilitate participation and completion. To ensure clarity and reasonable response time, the authors internally tested the questionnaire by simulating its completion and made adjustments to improve objectivity and response efficiency. Additionally, the draft was shared with two producers known to one of the authors to refine the final version. While no formal validation with a pilot group or external experts was conducted, the methodology ensured a balance between data relevance, response feasibility, and field applicability.
The classification of herd size concerning milk production, according to SANTOS & BITTAR (2015), determined small farms as those producing less than 200 liters/day; medium-sized farms, those producing between 201 and 700 liters/day; and large farms, those with production greater than 701 liters/day. Additionally, the farms were classified according to the type of animal production system, as described by LUCIANO et al. (2017), into extensive, intensive, or semi-intensive. With these classifications, it was possible to group respondents and establish appropriate statistical correlations for the sample group, to provide greater clarity to the results obtained.
With the results in hand, the data was coded and entered into a Microsoft Office Excel® spreadsheet and analyzed using the statistical software StataCorp/14 (2015). The data were exported to the program containing 91 viable respondents. The Shapiro-Wilk test was used to verify the normality of the quantitative variables related to milk production in liters. Comparisons between categorical variables (farm size and type of farming system) regarding the adoption of biosecurity practices were performed using Fisher’s exact test. Given the significant associations, pairwise comparisons were made by observing standardized residuals, with values less than or greater than the range -1.96 and 1.96 considered significant (DOHOO et al., 2012). For all analyses, a statistical significance of 5% was considered.
RESULTS AND DISCUSSION
Brazil still faces diseases in bovine herds that are significant for both animal production and public health, such as brucellosis and tuberculosis. Additionally, the country is in the process of becoming free from foot-and-mouth disease without vaccination. Advancing biosecurity practices is essential for improving animal health, which impacts the exportation of animal products. Although, biosecurity measures are essential in animal production systems, it is poorly implemented across dairy farms in Brazil, particularly in small operations that often lack such practices (FERREIRA et al., 2024). Thus, studies like this one can provide essential insights for dairy sectors in developing effective control strategies and offering guidance to cattle farmers on managing and preventing diseases.
Data were obtained from 91 dairy farms, with participation from the following Brazilian states: Minas Gerais (69 - 75.8%), Bahia (3), Ceará (2), Federal District (2), Espírito Santo (1), Goiás (3), Piauí (1), Rio de Janeiro (3), Rio Grande do Norte (1), Rio Grande do Sul (1), Santa Catarina (1), São Paulo (3), and Sergipe (1). The dichotomous questions and answers contained in the questionnaire are described, separated by external and internal biosecurity measures and subdivided according to the areas of the production system, analyzed according to the classifications of farm size and type of farming system.
Table 1 examines how external measures varied according to the farm size and table 2 focuses on differences in external measures conforming to the type of production system.
In this study, the use of technology and veterinary assistance were significantly correlated with larger farms and semi-intensive production models. This finding aligns with the study by DELA RUE et al. (2020), which surveyed 500 dairy farmers in New Zealand and showed that the use of technology is more common in dairy farms that typically have larger, newer herds and large-scale production. Use of technology and veterinary assistance provide better monitoring, early disease detection, and more precise herd management, which reduce disease outbreaks and improve overall animal health (BUSHRA et al., 2024). However, smaller farms often face financial and structural barriers to adopting such practices (DENIS-ROBICHAUD et al., 2019; BUSHRA et al., 2024). Expanding access to resources would improve the overall health of the dairy sector, reduce the risk of disease transmission, and promote sustainable practices across farms of all sizes. Additionally, as the next results will reinforce, herd size plays a significant role in determining the adoption of biosecurity practice.
DAMIAANS et al. (2019) evaluated the level of implementation of biosecurity practices in calf rearing on Belgian farms and found that measures related to animal movement were described as risk factors for several diseases and were among the most frequently cited by respondents. In the study in question, regarding the introduction of new animals, the practice of acquiring cattle from other farms was adopted by 55% of the properties. Although this practice was not associated with herd size or type of production system, the practice of testing for diseases when purchasing animals was associated with larger farms (Table 1). These results contrast with those of a recent study conducted by FERREIRA et al. (2024) on 69 dairy farms in the Campos Gerais region, in the state of Paraná, Brazil. In this study, the relationship between biosecurity measures and farm size was investigated, where the results showed that the purchase of animals was more common in medium and large farms, with 61% of medium-sized farms and 75% of large farms making acquisitions, compared to only 40% of small farms.
PEGORARO et al. (2018) assert that the best strategy to prevent the introduction of infectious diseases into the herd is to keep it closed, without introducing new animals. However, when purchasing is unavoidable, animals with a known disease history should be chosen, and before integrating them into the herd, they should undergo serological testing for major infectious diseases. According to table 1 and table 2, this practice was more common in larger properties. After purchasing the animals, 58% of respondents reported that they test for some disease, in contrast to the research by HARUN et al. (2022), where 40% of owners tested their animals before introducing them into the herd, and the study by FERREIRA et al. (2024), which revealed that only 29% of the analyzed farms conduct serological tests before introducing new animals, with this practice being more common in smaller properties.
Sixty percent of respondents stated that they conduct tests for Brucellosis and Tuberculosis, and this action was significant in larger properties and intensive production systems. According to the National Program for Brucellosis and Tuberculosis Control and Eradication (PNCEBT), recommended prevention methods include diagnostic testing and mandatory vaccination against brucellosis for all female cattle and buffalo aged 3 to 8 months. Only 11 participating farms (12%) do not have or do not follow the vaccination schedule, contrary to what was shown by DAMIAANS et al. (2019), where more than half of the farmers (65%) did not do so because they believed vaccination was not important, found it too expensive, or because most vaccines can only be administered at a certain age of the animal.
DENIS-ROBICHAUD et al. (2019) observed that, when associating the implementation of a quarantine plan with the sociodemographic characteristics of dairy farms in Canada, employees with higher levels of education found this practice to be more effective than those with lower levels of education. A similar result was found in the study by BUSHRA et al. (2024), which analyzed 205 dairy farms in five districts of Bangladesh. Educated farmers achieved better results in biosafety, health, and disease management practices compared to those who were illiterate or had only primary education. This difference can be explained by the tendency of farmers with lower educational levels to compromise biosafety standards.
When acquiring herds with unknown or doubtful health status is to maintain them under quarantine or separate them from other animals (PEGORARO et al., 2018). This practice was significantly attributed to intensive systems in the study, and only 9 respondents (18%) implemented it, approximately the same as reported by HARUN et al. (2022). A different result was identified by BUSHRA et al. (2024), found that among farmers who bought new cattle in the past year, 26.8% vaccinated them and 56.2% quarantined them before integrating them into the herd.
It’s evident that measures related to disease management, such as veterinary consultancy, vaccination, diagnostic testing, and quarantine of newly acquired animals, are typically linked to larger herds and intensive production systems in the study. A similar result was identified by DENIS-ROBICHAUD et al. (2019), where biosecurity measures associated with animal movement are commonly adopted in Canadian farms of medium to high production, likely due to practical and economic viability, but also due to a greater perception of risks and threats to dairy farming. Similarly, VILLARROEL et al. (2007), in their outline on critical control points for biosecurity in large dairy farms in the USA, noted that most respondents had an established vaccination protocol, which was reviewed annually by veterinarians.
The articles presented and our results highlight the significant role farm size and production model play in the adoption of biosecurity practices and veterinary assistance. Larger farms are more likely to invest in technology and veterinary services. However, smaller farms face substantial barriers to adopting these practices, primarily due to financial constraints, limited access to veterinary resources, and lower education levels. To address this gap, it is crucial to implement policies that provide financial support and training to smaller producers, enabling them to overcome these barriers, making biosecurity more accessible to smaller farms. Expanding access to diagnostic tools, veterinary guidance, and biosecurity education could lead to more uniform disease management practices, ultimately benefiting the entire dairy sector.
Access control to the interior of the property is a critical point regarding the biosecurity of dairy farms. In 63 participating properties (69%), people have unrestricted access to animal facilities, which contradicts DAMIAANS et al. (2019), who reported that in 65% of the properties, access was controlled by closed gates, with the requirement for visitors to announce themselves before entering. When asked about the conduct of employees upon arriving at the property, 28 owners (31%) require, as a property rule, bathing, changing clothes, and wearing clean boots. The same was described by DAMIAANS et al. (2019), where 30% of the properties provided boots, and six (20%) provided clean or specific clothing for visitors and technicians. Conversely, in the study by HARUN et al. (2022), only 5% of producers provided specific clothing for visitors. Similarly, the study by BUSHRA et al. (2024) highlighted that hygienic measures related to the use of clean coveralls and boots by animal caretakers, visitors, and agricultural service providers were largely neglected, with 98.5% of farms not adopting these practices. The lack of access control to dairy farm facilities and inadequate hygiene practices for workers and visitors present significant biosecurity risks, as unrestricted access can facilitate the introduction and spread of infections. Moreover, the limited adoption of hygiene measures, such as the provision of clean clothing and boots, further compounds these risks (MEE et al., 2012). Farm management should prioritize educating staff on the importance of hygiene and access control, as well as providing the necessary resources for their implementation.
Practices related to the care of pregnant cows and newborn calves are among the main ones in preventing the spread of existing pathogens within the herd. According to table 3, table 4, and table 5, 62% of respondents reported having a maternity pasture or facility for this category within the property, and the existence of this location was associated with medium and large properties and intensive production systems in the study. This finding contrasts with the research of HARUN et al. (2022), where 90% of properties do not have maternity barns, coupled with the fact that over 50% of the properties studied by him are low-income and have small herds and/or extensive management.
When asked about cleaning and disinfecting the maternity area after each birth or abortion, only 13% of respondents practiced this, and 12% did so only in cases of abortion. These results differed from a study conducted on farms in Quebec, Canada, where biosafety questionnaires were completed by 3,825 dairy producers. In that study, 47% of producers reported not having maternity areas, and among those who did, only 55% reported disinfecting and cleaning the pen after birth (LIMA-CAMPÊLO et al., 2024). PEGORARO et al. (2018) explained that areas designated for this purpose should always be clean, dry, well-ventilated, located near the pens, and easily visible, and that the cleaner and disinfected they are, respecting the sanitary void, the more effective the bio compartmentation and biocontainment will be (VILLARROEL et al., 2007; RENAULT et al., 2020).
According to table 3 and table 5, in half of the properties (51%), calves are separated from their mothers shortly after birth and housed in groups (54%) or individually (46%). Additionally, collective calf pens were significantly attributed to medium-sized farms and extensive production systems, while individual calf pens were associated with larger properties and intensive systems. The same was indicated by VILLARROEL et al. (2007) in their discussion on critical control points in the management of newborn calves on large dairy farms. In this case, it was noted that, typically, in these properties, calves are born in a maternity area and are soon separated from their mothers and transferred to individual areas. This practice minimizes the potential risks of pathogen transmission, especially those related to the gastrointestinal and respiratory tracts of the animals.
Questions regarding the management of colostrum for calves were not addressed; although, respondents were asked about the use of colostrum banks. There were associations between large and intensive farms performing colostrum management exclusively with a colostrum bank, and extensive farms with lower milk production using the natural colostrum method.
The results discussed above highlighted significant biosecurity challenges in the management of pregnant cows and newborn calves. Only a small proportion of the farms implement adequate cleaning and disinfection protocols, a critical biosecurity measure. Well-maintained, clean, and disinfected maternity areas are essential for controlling pathogen transmission. Furthermore, the practice of early separation of calves, while more common on larger, intensive farms, remains less consistent in smaller, extensive systems. Additionally, the management of colostrum, particularly through colostrum banks, is notably absent in most farms, further contributing to potential pathogen transmission risks (MEE et al., 2012). These findings underscore the need for improved biosecurity practices across all farm types, particularly in smaller and extensive operations, to reduce pathogen risks and enhance herd health. Integrating routine sanitation, better calf management, and colostrum control into farm practices is crucial for effective pathogen containment and overall biosafety.
Correct management during milking, both in mechanical and manual processes, requires special attention to biosecurity practices. Among the surveyed properties, 82% use mechanical milking, 80% clean and disinfect teats before milking (pre-dipping), and 74% disinfect teats after milking (post-dipping). The latter differed in extensive production systems. Additionally, all respondents clean or disinfect equipment at the end of milking. In contrast, the study by BUSHRA et al. (2024) found that on most farms (94.1%), milkers washed and sanitized their hands before milking. Teat dipping before and after milking was practiced in 3.4% of farms, with teat dipping before milking (pre-dipping) practiced in 0.5% and teat dipping after milking (post-dipping) practiced in 3.4% of farms only.
YANG et al. (2019), examining the adoption rate of biosecurity practices related to milk safety on dairy farms in northern China, found that teat antisepsis before milking was performed by 85.71% of properties, and 70.59% after milking, and that the average adoption rates of measures related to milk hygiene increased with the size of the property.
The implementation of biosecurity practices during milking is crucial for minimizing pathogen transmission and ensuring milk safety (JIMENEZ et al., 2023). Despite the high level of equipment sanitation observed, the variation in teat dipping practices between extensive and intensive systems indicates that biosecurity adoption is influenced by farm size and production system. While intensive systems tend to adopt these measures more consistently, smaller farms may face barriers such as limited resources or lack of knowledge, as seen in the lower rates of practice in extensive systems. These findings highlighted the need for targeted biosecurity training, especially for smaller and extensive farms, to ensure uniform implementation of critical hygiene practices and minimize risks to herd health.
Regarding issues related to preventing and maintaining the hoof health of the herd, as shown in table 4 and table 6, just over half of the respondents (56%) perform hoof trimming of the animals, and only 35% have a routine of disinfection through footbaths. These practices were significant in properties with medium to high milk production and in intensive models. According to HASKELL et al. (2006), confined and semi-confined systems, characteristic of intensive high-production farms, face the risk of increasing lameness and knee injury in their cows. Therefore, to ensure the welfare and hoof health of the animals, producers can address the negative effects of these conditions preventively, with treatments such as corrective hoof trimming and the use of footbaths, in addition to monitoring and improving the facilities and paddocks that accommodate these animals.
Just like the adoption of disinfection protocols, the non-sharing of materials with other properties, such as syringes and needles, constitutes biocontainment practices (RENAULT et al., 2020). Observing table 4 and table 6, eighty-six percent of the properties in the study do not share materials; however, needles are multi-use (73%). Notably, 71% of respondents disinfect needles before reusing them. This suggested a need for stricter biosecurity measures, as reusing needles, even when disinfected, still poses a risk for pathogen transmission.
Regarding waste management, in Brazil, there is still no legislation regarding the disposal of dairy cattle waste in the soil, despite its widespread use under agronomic recommendations, such as fertilizer (PEGORARO et al., 2018). In this study, the collection and storage of manure are carried out by most properties (74%); although, only 9 (10%) subject it to any type of treatment, which was higher than described by HARUN et al. (2022), where 65% of properties cleaned and disinfected the stall after manure removal. As treatment alternatives, respondents described the use of biodigesters, manure pits, and compost bins.
Finally, for the prevention and control of infectious diseases, the existence of a space dedicated to the disposal of carcasses was observed on 63 properties (69%), protected from access by other animals. Respondents revealed that carcasses were buried, incinerated, disposed of in pastures and/or trenches, or deposited in compost bins. This percentage was lower than that detected in the work of DENIS-ROBICHAUD et al. (2019), where 92% of properties adopted the same disposal methods, including storage and freezing of carcasses. Additionally, as demonstrated by HARUN et al. (2022), carcasses were delivered to specialized services.
Once again, waste management practices were significant in larger properties and intensive production systems. According to the assessment by GWYTHER et al. (2011), concerning the socioeconomic and biosecurity aspects of methods used worldwide for carcass disposal, burial, burning, and composting offered the best cost-benefit ratio and practicality for farmers. Moreover, it is ensured that more sophisticated practices, such as on-farm or large-scale incineration, become profitable with increased production and herd size.
The main limitation of this study is the participation rate and the absence of follow-up on the participants. Researchers have tried different methods to improve response rates, such as monetary and non-monetary incentives, changes in the survey length, mode of administration, and appearance of the questionnaire (SMITH et al., 2019; BOOKER et al., 2021; LAVIDAS et al., 2022). Our study was voluntary and in Brazil is not allowed monetary incentives. Before the application of the questionnaire, we tested it with some dairy farms to adequate the length. Possible reasons for the low response rate in our study may include concerns about privacy and confidentiality (despite assurances in the consent letter), lack of interest, low engagement, high demanding work schedules, age, and internet (SMITH et al., 2019; BOOKER et al., 2021). Additionally, one study revealed that the respondents are more likely to participate in survey when the web survey sponsorship has a reputation or is known to the participants (LAVIDAS et al., 2022). Moreover, cultural characteristics of sector should be considered, and if the target population do not find the research relevant, it could significantly reduce response rate (BOOKER et al., 2021).
Although, this study had a limited number of respondents, it offers a significant contribution to understanding biosecurity practices in dairy farms in Brazil. Unlike studies that aimed to broadly characterize properties or provide a detailed explanation of biosecurity application, this work focuses on revealing, from the perspective of the questionnaire respondents, which specific measures are being implemented. Therefore, the goal is not to generalize the results to the entire population of dairy farms but to provide an overview of the biosecurity practices reported by those who participated in the survey.
CONCLUSION
The study highlighted a positive perception among livestock farmers regarding the importance of implementing biosecurity practices within farms, despite their still low adoption rates in the investigated properties, particularly in small and medium-sized farms and extensive production systems. However, even in these properties, critical practices are followed, such as animal vaccination schedules and those related to fomites and visitors.
Understanding how producers utilize biosecurity, identifying which measures they find useful, and factors that can potentially influence their acceptance are crucial. This understanding should be based on individual property characteristics and potential risks. Additionally, all stakeholders involved in the sector must be aware of the risks associated with the introduction and spread of pathogens within the property. This shared awareness will foster mutual efforts towards biosecurity measures that contribute to establish a production system grounded in herd health and quality.
ACKNOWLEDGEMENTS
We would like to thank all milk producers in Brazil who collaborated with this research by answering the questionnaire; Fundação Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and IFMG for encouraging research and for granting scholarships to undergraduate research students.
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Rudi Weiblen (0000-0002-1737-9817) Rudiger Daniel Ollhoff (0000-0002-7972-9472)
