Assessment of reported vaccination coverage against neurotropic infectious diseases in equines from the Vale do Paraíba, BrazilABSTRACT
Infectious neurological diseases of the equine central nervous system are major health concerns, and vaccination is the main preventive measure. This study assessed vaccination coverage against rabies, encephalomyelitis, tetanus, and type I equine herpesvirus myeloencephalopathy in horses from the Vale do Paraíba region (São Paulo, Brazil). A cross-sectional survey with face-to-face questionnaires was conducted on 30 properties, covering 1,140 animals. Sampling followed the WHO-adapted 30-by-7 method, and analyses included principal component and multiple correspondence analysis. Most horses were adult females used in equestrian sports. The vaccination rates were 95% for rabies, 84% for encephalomyelitis and tetanus, and 14% for herpesvirus. Sport horses had higher vaccination adherence, while working horses showed lower coverage. On properties with foals, about 20% did not follow manufacturer recommendations for herpesvirus vaccination. The main study limitation was the absence of a regional equine census and centralized data. Although some properties showed high vaccination rates, others had clear immunization gaps. These findings can help guide regional vaccination strategies and inform similar initiatives elsewhere.
Keywords:
horse; epidemiology; vaccine
RESUMO
As doenças neurológicas infecciosas que acometem o sistema nervoso central de equinos representam importantes preocupações em saúde, e a vacinação é a principal medida preventiva. Este estudo avaliou a cobertura vacinal contra raiva, encefalomielite, tétano e mieloencefalopatia por herpesvírus equino tipo I em cavalos da região do Vale do Paraíba (São Paulo, Brasil). Foi realizado um inquérito transversal com questionários presenciais em 30 propriedades, abrangendo 1.140 animais. A amostragem seguiu o método 30-por-7 adaptado pela OMS, e as análises incluíram análise de componentes principais e análise de correspondência múltipla. A maioria dos equinos era composta por fêmeas adultas utilizadas em esportes equestres. As taxas de vacinação foram de 95% para raiva, 84% para encefalomielite e tétano, e 14% para herpesvírus. Cavalos de esporte apresentaram maior adesão vacinal, enquanto equinos de trabalho mostraram menor cobertura. Em propriedades com potros, cerca de 20% não seguiam as recomendações do fabricante para a vacinação contra herpesvírus. A principal limitação do estudo foi a ausência de um censo equino regional e de dados centralizados. Embora algumas propriedades apresentassem altas taxas de vacinação, outras evidenciaram lacunas claras de imunização. Esses achados podem contribuir para orientar estratégias regionais de vacinação e subsidiar iniciativas semelhantes em outras localidades.
Palavras-chave:
cavalo; epidemiologia; vacina
INTRODUCTION
Neurological diseases of infectious origin affecting the central nervous system of horses, including viral encephalomyelitis, rabies, herpesvirus myeloencephalitis and tetanus, are recurrent and particularly relevant in specific geographic regions (Costa et al., 2015). Treatment for some of these conditions has limited effectiveness, while vaccination remains the primary preventive measure (Chagas et al., 2019). Due to the economic importance of equines in the region (Sachs et al., 2006), the establishment of epidemiological surveillance becomes essential to preserve both animal and human health. Vaccination, combined with early detection and disease monitoring, supports more effective prevention and control policies.
The Vale do Paraíba region is in the southeast of the state of São Paulo and comprises thirty-nine municipalities distributed into five sub-regions (Fig. 1) (Mutuzoc and Vieira, 2012). Despite the global relevance of neurotropic infectious diseases in equines and the recognized need for effective preventive measures, regional information on equine health management and vaccination practices remains scarce. The region has a Gross Domestic Product (GDP) of R$ 28,500,000,000 (January-March 2019), equivalent to approximately US$ 7.5 billion at that year's exchange rate, representing the second-highest economic output among the 15 metropolitan and administrative areas of the state of São Paulo (Pib…, 2019). Agribusiness plays a central role in this economic profile, and the equine industry alone generates six times more jobs than the automobile sector in Brazil, accounting for nearly three million positions (Brasil, 2016). However, the absence of updated, region-specific data on preventive practices highlights an important gap, especially considering the economic and social relevance of equine production in the Vale do Paraíba.
The diseases included in the study differ in terms of risk, epidemiology, and zoonotic potential, but all have significant implications for horse breeding. Literature recommends vaccination and classifies infectious diseases in three groups, as follows. Group 1 includes rabies, tetanus, and viral encephalomyelitis, and vaccination against these diseases is considered essential (it is strongly recommended), as they pose a high risk of animal mortality, have a major impact on public health, and result in economic losses. Group 2 includes herpesvirus myeloencephalopathy and equine influenza, and immunization is only recommended. Group 3 includes other diseases not covered in this study (Ferraz, 2014). Vaccination is highlighted as the most efficient and economical strategy for herd protection. There are no data available about epidemiological surveillance or vaccination programs for the prevention and control of neurotropic diseases in horses of the Vale do Paraíba. Therefore, we aimed to estimate the vaccination coverage against rabies, encephalomyelitis, tetanus and herpesvirus type I myeloencephalopathy in horses from the Vale do Paraíba region (São Paulo, Brazil), determining the sampling profile, vaccination frequency, evasion, and adherence to protocol recommendations.
ETHICAL ASPECTS
The research was submitted to the Ethics Committee on Animal Use of the University of Vale do Paraíba (UNIVAP), and approved under the number CEUA03/2021.
MATERIALS AND METHODS
The “30 by 7” method published by the Department of Immunization, Vaccines, and Biologicals - World Health Organization (Hoshaw-Woodard, 2001) was adapted to define the sample size of this study. The adaptation consisted of selecting 5 clusters (5 municipalities, representing the 5 sub-regions selected aiming for greater representativeness) and, within these, 5 units (5 different types of properties, aiming to better represent the actual population in the sample profile). The adaptations in the number of clusters and units were made based on practical feasibility. The Vale do Paraíba region is divided into five sub-regions, each of them comprising various municipalities were selected through non-probabilistic sampling based on the highest Gross Domestic Product (GDP) in each of the sub-regions: São José dos Campos was defined as the representative of sub-region 1 (SR1), Taubaté for sub-region 2 (SR2), Guaratinguetá for sub-region 3 (SR3), Cruzeiro for sub-region 4 (SR4), and for sub-region 5, we chose two municipalities - Ilhabela and São Sebastião (Fig. 1). Within each municipality, an inventory of the properties it contained was carried out through local contacts, and invitations to participate in the study were sent. From those that accepted, a random selection was carried out to choose one representative of each property type established as the selection criterion. We defined 'properties' the six most common types of facilities in this region to be evaluated in each of the municipalities selected (Horse Farms, Riding Clubs, Training Centers, Farms, Ranches, Small Farms). The criterion for including a unit (property) in the study was: properties with horses falling into any of the classifications used in this study, provided they agreed to participate following prior telephone contact. All animals present on each property were included, in accordance with the methodology previously described in this study.
Vale do Paraíba map, São Paulo, Brazil (2022) and its sub-regions with the selected municipalities for this study.
A questionnaire developed by the authors was administered by a qualified team member through in-person interviews, following the signing of the Informed Consent Form, and conducted with individuals responsible for the animals (either the animal owner or the property manager). The questionnaire included 83 structured questions covering the following topics: property identification (identification code, municipality, whether there was a responsible veterinarian, the main breed raised, and the purpose of the animals); and vaccination (the specified number of vaccinated adult horses and foals, males and females; whether they were vaccinated against the diseases under study; whether the vaccinations were up to date; which application protocol was used; and the brand of vaccine administered). Most of the questions were open-ended, allowing for the collection of the most detailed and relevant information possible. This approach was adopted due to the absence of official documents and records that would allow for the collection of information in a more standardized and reliable manner. The instrument used in this study was not formally validated, but it was pretested with a pilot group to assess item clarity, comprehension, and relevance. Feedback from the pretest was incorporated into the final questionnaire.
The minimum vaccination coverage rate recommended by the WHO and the Brazilian Ministry of Health to consider a population protected against rabies is 80%. As there are no specific regulations for horses or for the other diseases addressed in this study, we extrapolated this recommended value (Kotait and Gonçalves, 1982; Schneider et al., 1996; Dia…, 2019)
Statistical analyses were performed to evaluate associations among vaccine types and to explore patterns in the relative percentage of vaccinated animals. A 5% significance level was adopted. All procedures were conducted in R (Version 4.1.0) within RStudio, with functions cited as ‘package::function’.
Principal component analysis (PCA) was applied to the relative percentage of animals vaccinated for each disease to identify underlying multivariate structures. The number of retained dimensions was determined by Horn’s parallel analysis, and four biplots were generated to illustrate these patterns (stats::princomp; psych::fa.parallel).
Multiple correspondence analysis (MCA) was used to examine interdependencies among sub-regions, management functions, animal categories, breeds, vaccination scores, and vaccine types. Vaccination percentages were converted into qualitative categories (>80% or <80%), and the data matrix was transformed into a Burt table for visualization in a two-dimensional perceptual map (FactoMineR::MCA; GDAtools::burt; factoextra::fviz_mca_biplot).
The Burt table underwent chi-squared testing to extract residuals, which were standardized using the z-normal scale. Interdependencies were considered significant when |Z| > 1.96, following Agresti (2007). Technical procedures used stats::chisq.test.
To define property profiles, individual scores from the first two MCA dimensions were extracted and compared through hierarchical clustering based on Euclidean distance and the Ward D method, revealing two major clusters. The same scores were also classified using K-means into two groups, incorporated into a new Burt table, and reanalyzed via standardized residuals (factoextra::get_mca_ind; stats::dist; stats::hclust; stats::kmeans).
RESULTS
The data described here were obtained through responses to questionnaires administered via interviews. It is important to emphasize that both the sample profile and the vaccination frequency, as well as adherence to recommendations, are entirely dependent on the report of the person responsible for the animals, due to the absence of official records.
Data concerning 1,140 animals were obtained from 30 different properties located in the five sub-regions of Vale do Paraíba. The proportional distribution of animals per sub-region was: 26% (N=298) for SR1, 37% (N=425) for SR2, 17% (N=192) for SR3, 11% (N=124) for SR4 and 9% (N=101) for SR5. The overall distribution of the population according to the type of property in this study was: 47% (N=531) Horse Farms, 20% (N=231) Riding Clubs, 10% (N=117) Training Centres, 7% (N=77), Farms, 13% (N=151) Ranches, and 3% (N=33) Small Farms. The overall demographic profile of the population can be seen in Table 1.
Demographic distribution of the horses in the sample studied across the five sub-regions of the Vale do Paraíba, São Paulo, Brazil, in 2022, according to breed, sex, age group (foals and adults), and activity performed (sport, leisure, or work). The values are presented as absolute numbers and percentages within the total evaluated (n = 1,140)
Vaccination coverage for the diseases considered in this study are listed in Table 2.
Distribution of the absolute and relative numbers (based on the total study population, n=1,140 animals) of horses vaccinated against rabies, encephalomyelitis, tetanus, and viral meningoencephalitis, according to the five sub-regions of the Paraíba Valley, São Paulo, Brazil.
Distribution of the percentage of horses vaccinated against rabies, encephalomyelitis, tetanus, and viral myeloencephalopathy across the five sub-regions of the Vale do Paraíba, São Paulo, Brazil, in 2022 (Fig. 2). The thematic maps illustrate spatial variations in vaccination coverage, with percentage gradients represented by the color scale (according to the legend in the center of the figure, where each color corresponds to a specific percentage range) (n=1,140).
Percentage distribution of horses vaccinated against rabies, tetanus, encephalomyelitis, and viral myeloencephalopathy, stratified by category: total sample, adults, males, females, pregnant mares, and foals, in the Vale do Paraíba, São Paulo, Brazil, in 2022 (Fig. 3). The charts show the proportion of adherence (Yes - vaccinated animals) and non-adherence (No - unvaccinated animals) to vaccination for each disease in each group. (N=1,140).
Percentage of vaccinated animals against neurotropic infectious diseases in the different sub-regions of the Vale do Paraíba, São Paulo, Brazil, 2022.
Percentage of horses vaccinated against rabies, tetanus, encephalomyelitis, myeloencephalopathy, in each category. Vale do Paraíba, São Paulo, Brazil, 2022.
Only the first Principal Component (PC) was retained according to Horn’s parallel analysis, with PC1 explaining 59.82% of the total variance. The relative vaccination percentages for rabies, encephalomyelitis, and tetanus loaded onto PC1 (loadings>|0.40|), indicating positive associations among these diseases. Encephalomyelitis and tetanus showed particularly strong correlation (Table 3). The myeloencephalopathy vaccine did not load onto PC1, likely due to its low frequency of administration. This pattern indicates that properties tend to vaccinate (or fail to vaccinate) consistently across these three diseases, reflecting a unified management style rather than disease-specific decision-making.
Factor loadings of the variables evaluated in the first three principal components (PC1, PC2, and PC3), accompanied by the eigenvalues, the variance explained by each component, and the cumulative variance resulting from the principal component analysis (PCA). Bold values indicate factor loadings considered relevant (>|0.40|), used to interpret the contribution of each variable to the formation of the components.
Qualitative assessment of the biplot revealed clear differences among functions, sub-regions, property types, breeds, and veterinary care regarding vaccination coverage (Fig. 4). Working horses were positioned farther from vaccine vectors, suggesting lower vaccination rates, whereas sport horses were closer to these vectors, indicating higher coverage (Fig. 4A). This suggests that horses used for labor may receive lower preventive care due to economic or management limitations, while sport horses - typically considered of higher commercial value - are consistently vaccinated.
Sub-regions showed similar centroid positions, except sub-region 1, which was closer to the myeloencephalopathy vaccine, indicating comparatively higher vaccination rates for this disease (Fig. 4B). This may reflect localized awareness campaigns, higher disease pressure, or greater access to veterinary services within this sub-region.
Ranches were located farther from vaccine vectors, suggesting lower vaccination coverage in this property type (Fig. 4C). This pattern indicates that ranch-type operations may have structural or operational barriers that reduce adherence to vaccination protocols.
Brazilian Horses and Quarter Horses were positioned closer to the myeloencephalopathy vector, indicating greater uptake of this vaccine among these breeds (Fig. 4D). This likely reflects selective prioritization of these breeds, which are often used in competitive or high-value activities where vaccination is more rigorously followed.
Finally, properties receiving veterinary assistance were closer to vaccine vectors, demonstrating higher overall vaccination coverage (Fig. 4E). This finding reinforces the importance of professional veterinary involvement as a determinant of adequate vaccination practices.
Distribution of the relative percentage (based on the PCA) of horses vaccinated against each disease in reference to functions, sub-regions, type of property, breed and veterinary assistance. Vale do Paraíba, São Paulo, Brazil, 2022.
Distribution of the relative vaccination percentages of horses for each disease based on Principal Component Analysis (PCA), in reference to the different categories analyzed: (A) functions; (B) sub-regions; (C) type of property; (D) breed; and (E) veterinary assistance. Smaller circles represent individual observations, whereas larger circles represent the centroids of each evaluated category. Centroids correspond to the center of gravity formed by interpolating the points of the same color. Arrows indicate the vectors of the variables (diseases), demonstrating their influence on the main axes (Dim1 and Dim2). Vale do Paraíba, São Paulo, Brazil, 2022. (%) = Relative number.
Here we demonstrate the difference between what is done and what is recommended by vaccine manufacturers. In Fig. 5, we present the adherence and non-adherence to the recommended protocol found in our survey according to each disease in each group.
Percentage distribution of properties that followed or did not follow the vaccination protocol recommended by the manufacturer for rabies, tetanus, encephalomyelitis, and viral myeloencephalopathy, stratified by animal categories (adults, pregnant mares, and foals) in the Vale do Paraíba, São Paulo, Brazil, in 2022. The charts show the proportion of adherence (Yes - properties that followed the protocol) and non-adherence (No - properties that did not follow the protocol) to the manufacturer-recommended vaccination protocol for each disease in each group. (N=1,140).
Percentage of properties that adhered or did not adhere to the protocol recommended by the vaccine manufacturer from the Vale do Paraíba, São Paulo, Brazil, 2022.
It was possible to identify response patterns through the analysis of standardized and adjusted z-normal residuals from the MCA (Table S1), which revealed interdependence among variables. Properties that failed to follow the manufacturer-recommended protocol for one vaccine generally did not follow the protocols for the other vaccines, even when more than 80% of their herds were vaccinated (Table S2). Conversely, properties adhering to the manufacturer’s guidelines for one vaccine tended to do so for all vaccines (Table S3). This suggests that non-compliance is not linked to a specific vaccine, but rather to the owner’s overall profile regarding adherence to vaccination recommendations.
In the adult horses and pregnant mares categories, at least 50% of properties housing these groups followed the manufacturers’ recommended protocols. In contrast, fewer than 30% of properties with foals complied with the recommended protocols for this category (Figure 5). Although a general pattern of adherence or non-adherence exists, foals appear to be the most affected by protocol non-compliance. This lower adherence in foals may reflect management challenges, lower prioritization of young animals, or limited knowledge about age-specific immunization requirements.
Based on the individual MCA scores of the first two dimensions, hierarchical clustering identified two groups of properties (Fig. 6). The interdependencies revealed by the adjusted and z-normalized residuals indicated that Profile 1 comprised properties with higher vaccination coverage, whereas Profile 2 consisted of properties with lower vaccination coverage (Table S4 and Figure S1). These two clusters reinforce that properties tend to exhibit consistent vaccination patterns, forming distinct profiles of high and low compliance that may guide targeted intervention or education strategies.
Individual scores of the first two dimensions of the multiple correspondence analysis showing the formation of two clusters in the sample from the Vale do Paraíba, São Paulo, Brazil, 2022.
Graphical representation of the two major clusters identified by the MCA (Profile 1 and Profile 2) among the studied properties (numerically represented on the X-axis) based on the dissimilarity within the population (Y-axis) from Vale do Paraíba, São Paulo, Brazil, 2022. These profiles are composed of individuals whose categorical responses/variables are more similar within each group and more distinct between groups.
DISCUSSION
After checking local official documents, it became evident that vaccination is mandatory only against influenza and for animals taking part in equestrian events and crowded gatherings. However, no mandatory requirements exist for other diseases, including those classified in Group 1, which are theoretically more severe than Group 2 (where influenza is included). Organizations promoting equestrian events may require, on their own, vaccines included in the groups mentioned above (Brasil, 2001; Ferraz, 2014; Defesa..., 2021; Regulamento…, 2022; Veterinary, 2023). This regulatory gap suggests insufficient understanding of the immunization needs of these populations, which constitutes the objective of the present research.
There are several methodologies to assess vaccination coverage, and field surveys are widely accepted due to the high reliability of the data obtained. Among these, the 30 by seven method stands out for its 95% reliability. As discussed by several authors, biases may occur that distort the assumptions presented in the results. Information bias is inherent, since the reported vaccination status depends entirely on the caretaker, and selection bias may occur due to the exclusion of subpopulations from the sampling frame.
One of the main difficulties encountered is the use of non-probability sampling and the lack of standardized and well-documented quality control procedures, which may reduce the reliability of results. Furthermore, the cost and time associated with large samples can be challenging, and the availability of vaccination documentation directly affects data accuracy (Cutts et al., 2013; Eisele et al., 2013; Cutts et al., 2016). Despite these limitations, field surveys remain valuable for describing demographics and vaccination characteristics of the population studied (Guérin, 1998; Hoshaw-Woodard, 2001).
The predominant demographic profile consisted of animals in horse farms, mostly adult females used in sports riding. This profile reflects only the study sample and not necessarily the entire regional equine population, due to sampling limitations. However, it allows greater precision in interpreting the obtained data. Because many animals were pregnant mares and foals, immunization management requires particular attention, especially considering their future participation in events involving large gatherings.
Based on these characteristics, vaccination plays a crucial role in protecting this group. Previous studies indicate that such factors increase infection risk (Hayama et al., 2010; Ferreira and Torman, 2013). According to the official immunization programs from the World Health Organization and Ministry of Health, a minimum vaccination coverage of 80% is necessary for population-level protection (Cutts et al., 2016). In our study, we observed satisfactory rates for rabies, encephalomyelitis and tetanus, suggesting likely high coverage, although limitations prevent confirmation. Percentages for myeloencephalopathy were below 80%, indicating likely low coverage.
Although reasons for non-vaccination were not investigated, low awareness among caretakers was evident, especially regarding myeloencephalopathy, which contrasts with the greater recognition of diseases relevant to human health, such as rabies (Dubé et al., 2013; Cutts et al., 2016; Monte, 2021). Studies show that about 69% of veterinary professionals do not know how to fulfil mandatory disease reporting raising concerns about under-recognition and, consequently, under-vaccination (Brasil, 2019; Dia…, 2019; Monte, 2021).
An interesting finding was that vaccination rates against viral encephalomyelitis and tetanus were similar. This association occurred because all vaccinated animals received polyvalent vaccines (Aguiar, 2012). When analyzing the principal components, the loading value of 0.96 confirmed this association. Reports indicated that caretakers purchase commercial formulations without full awareness of all diseases covered, which may encourage or discourage adherence depending on cost and accessibility. (Martins et al., 2012).
Despite high vaccination rates in some properties, the proportion of unvaccinated animals remains concerning, as it compromises herd immunity. Ranch animals and working equines exhibited the lowest coverage, indicating higher susceptibility (Hayama et al., 2010; Paillot et al., 2017). The risk increases in environments with crowding or frequent movement of animals (Ferreira and Torman, 2013). These findings highlight the need for targeted actions to increase vaccination in vulnerable groups (Hayama et al., 2010; Ferreira and Torman, 2013; Paillot et al., 2017).
In addition to the susceptible populations mentioned, even vaccinated animals may not be adequately immunized (Knight-Jones et al., 2014). We found that 18.7% of properties did not comply with vaccination protocols, particularly affecting foals, which often missed the primary sequence and boosters. Early disruptions compromise immunity duration and increase susceptibility (Paillot et al., 2017). Multiple factors influence protection levels, including vaccine characteristics, pathogen features, environmental exposure and host factors (Knight-Jones et al., 2014; Paillot et al., 2017). Furthermore, this study evaluates only vaccination coverage-not efficacy-so reported vaccination does not necessarily translate to immunological protection (Knight-Jones et al., 2014; Paillot et al., 2017; Chagas et al., 2019).
CONCLUSION
The study provided an estimate of vaccination coverage against the main neurotropic viral infectious diseases in horses in the Paraíba Valley. This enabled the identification of gaps in the protection of the equine population under study, where, considering the zoonotic potential of the diseases investigated, it is essential to implement more efficient vaccination programs, awareness-raising actions and continuous health monitoring. However, even with some values that suggest a likely high coverage, we also saw that it may not reflect the real immunization status of individuals due to the conditions in which the vaccines are administered. We emphasize the limitations inherent to the methodology of the present study, but it has potential for application, not only because of the relevance of the topic, but also because of the gap that was possible to highlight in this aspect of animal and human health. Future studies with larger samples and, if possible, through the evaluation of individual titers are necessary to obtain more solid conclusions and guide more effective actions.
ACKNOWLEDGEMENTS
We are grateful to Prof. Sandra Maria Fonseca da Costa, Ph.D. Researcher at the University of Vale do Paraiba (UNIVAP) Research and Development Institute (IP&D), for her contribution on the geographic study; to Prof. Regina El Dib (in memoriam), Ph.D. Researcher at University Estadual Paulista (UNESP) for guiding us through the universe of "Evidence-Based Health"; to Anna Celina de Moraes Alves e Freitas, for her exceptional contribution to our study with her deep knowledge of the English language; to Prof. Luisa Lina Villa, Ph.D. Researcher at the University of São Paulo (USP) and Dr. Fabio Luiz Tuna Vieira, for the encouragement and support that was opportunely provided to us, them both being, beyond inspirations, great examples of professionals whose assistance we had the honor of receiving.
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DATA AVAILABILITY STATEMENT
The research data are available in the repository <https://repositorio.univap.br/home>.
The research data are available in the repository <https://repositorio.univap.br/home>.
Map of the Vale do Paraíba region, in the state of São Paulo (SP), Brazil, depicting its territorial division into five administrative sub-regions (Subregion 1 to Subregion 5). The map highlights the municipalities selected for the study - São José dos Campos, Taubaté, Guaratinguetá, Cruzeiro, and Caraguatatuba - represented with different shades of gray to distinguish each sub-region, along with contour lines to identify municipal boundaries (the internal legend specifies the correspondence between the shades of gray and the sub-regions). The areas surrounding the Vale do Paraíba include the bordering states of Minas Gerais (MG) to the north and Rio de Janeiro (RJ) to the east. The orientation arrow at the bottom of the map indicates true north. Source: Prepared by Laboratório de Estudos das Cidades (Laboratory of Cities Studies) - UNIVAP (2022), based on IBGE (Brazilian Institute of Geography and Statistics) data.
