RESUMEN

Equine Piroplasmosis (EP) and Equine Granulocytic Anaplasmosis (EGA) are diseases that affect horses, transmitted by ixodid ticks, causing a nonspecific febrile syndrome. Equine Piroplasmosis is endemic in Brazil, and most horses are in enzootic stability. Serological and molecular studies carried out on horses in Brazil have shown the presence of Anaplasma phagocytophilum, however, the clinical relevance of this infection has not yet been established. The present study aims to evaluate the importance of Babesia caballi, Theileria equi, and A. phagocytophilum as etiological agents in horses with clinical manifestations suggestive of these diseases in the metropolitan mesoregion of Rio de Janeiro. A total of 45 animals with clinical signs were submitted to DNA extraction followed by qPCR test. Anaplasma phagocytophilum, Neorickettsia risticii and Theileria haneyi were not found in any of the horses with clinical signs, however 62.2% were infected with at least one agent of EP. Theileria equi was the most frequent etiologic agent (35.5%), followed by coinfection (15.5%) and B. caballi (11.2%). These results suggest that A. phagocytophilum has minor clinical importance in the region, while EP is frequently found in symptomatic horses, representing an important differential diagnosis in suspected cases.

Asunto(s)

Anaplasma phagocytophilum , Babesia , Babesiosis , Ehrlichiosis , Enfermedades de los Caballos , Theileria , Theileriosis , Caballos , Animales , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/epidemiología , Brasil/epidemiología , Theileria/aislamiento & purificación , Babesia/aislamiento & purificación , Anaplasma phagocytophilum/aislamiento & purificación , Theileriosis/epidemiología , Theileriosis/parasitología , Babesiosis/epidemiología , Babesiosis/parasitología , Ehrlichiosis/veterinaria , Ehrlichiosis/epidemiología , Ehrlichiosis/microbiología , Masculino , Femenino

RESUMEN

Endometritis is the leading cause of mare subfertility. Most mares respond to standard therapy, but alternative therapies have been developed for mares failing to respond. This study aimed to investigate a commercially available, yet unassessed, product labeled as a uterine sanitizer to determine the in vitro antimicrobial activity against microorganisms associated with endometritis and its in vitro stability to dilute antibiotics. In experiment 1, the microdilution broth technique and antimicrobial effects were assessed against Escherichia sp, Staphylococcus sp., Klebsiella sp., Pseudomonas sp., and Candida sp. Percentage inhibition was calculated by comparing the optical density. The minimum inhibitory concentration (MIC) 100% was determined using the resazurin dye technique. MIC 50% and 90% were determined using a dose-response non-linear regression. In experiment 2, the uterine sanitizer was used to dilute commonly used antibiotics achieving a final volume of 90 mL at 5°C, 21°C, and 37°C. The pH was measured at 0, 1, 3, 6, and 24 h after dilution. The uterine sanitizer had inhibitory properties against all microorganisms; Escherichia sp. being the most susceptible, and Pseudomonas sp. the most resistant. The uterine sanitizer had an acidic pH=4; however, when combined with the antibiotics, the pH of the antibiotic remained unchanged with the different temperatures and did not precipitate. In conclusion, the uterine sanitizer showed antimicrobial effects against endometritis-causing microorganisms. The dilution of antibiotics in the uterine sanitizer was stable and this association could potentiate the antimicrobial effects. Uterine sanitizer's safety and clinical efficacy in vivo remain to be tested.

Asunto(s)

Antibacterianos , Bacterias , Endometritis , Enfermedades de los Caballos , Pruebas de Sensibilidad Microbiana , Femenino , Animales , Endometritis/tratamiento farmacológico , Endometritis/microbiología , Endometritis/veterinaria , Caballos , Antibacterianos/farmacología , Antibacterianos/química , Enfermedades de los Caballos/tratamiento farmacológico , Enfermedades de los Caballos/microbiología , Bacterias/efectos de los fármacos , Estabilidad de Medicamentos

RESUMEN

Leptospirosis is an anthropozoonosis of economic and public health importance, caused by bacteria of the genus Leptospira. Horses are deemed important in its transmission chain due to their proximity to humans, and because the species is often asymptomatic, making these animals potential silent reservoirs. In this context, the objectives of this study were to determine the prevalence of seropositive horses for Leptospira spp., and to identify the presence of Leptospira spp. serogroups and antibody titers, the occurrence of areas with higher density of infection cases and demographic characteristics associated with seropositivity in the states of Paraíba (PB), Pernambuco (PE), Rio Grande do Norte (RN) and Ceará (CE), in the Northeast region of Brazil, during rainy (May and June) and dry (October and November) seasons from 2017 to 2019. Using the microscopic agglutination test (MAT), 1152 equine serum samples from 225 municipalities were analyzed. Anti-Leptospira antibodies were detected in 23.9 % (95 % CI= 21.4 - 26.3 %) of the samples in the three-year period, with a frequency of 30.4 % (95 % CI= 26.7 - 34.2 %) during the rainy period (with greater emphasis on the Ballum serogroup) and 17.4 % (95 % CI= 14.3 - 20.5 %) in the dry period (with greater emphasis on the Sejroe serogroup). Age of horses ≥ 6 years (6-10 years, 11-15 years and ≥ 16 years), rainy season, and animal belonging to Pernambuco state were factors with higher seropositivities. Regarding spatial distribution, a higher percentage of seropositive animals was observed in Pernambuco (P < 0.05), in interstate border areas, and large urban centers, with a spatial cluster detected in the dry season of 2018 with relative risk of 2.8 (P = 0.049) times higher in municipalities within the cluster. It is suggested that measures for controlling rodents and contact with wild animals in equine farming, both in rainy and dry periods, combined with care regarding the use of pastures shared with cattle and the adoption of immunoprophylaxis are important in preventing and controlling leptospirosis in horses in the Northeast region of Brazil.

Asunto(s)

Enfermedades de los Caballos , Leptospira , Leptospirosis , Estaciones del Año , Animales , Caballos , Leptospirosis/veterinaria , Leptospirosis/epidemiología , Leptospirosis/microbiología , Brasil/epidemiología , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/microbiología , Leptospira/aislamiento & purificación , Leptospira/inmunología , Estudios Seroepidemiológicos , Prevalencia , Masculino , Femenino , Anticuerpos Antibacterianos/sangre , Análisis Espacial , Serogrupo

RESUMEN

Probiotic microorganisms can stimulate an immune response and increase the efficiency of vaccines. For example, Bacillus toyonensis is a nonpathogenic, Gram-positive bacterium that has been used as a probiotic in animal supplementation. It induces immunomodulatory effects and increases the vaccine response in several species. This study aimed to evaluate the effect of B. toyonensis supplementation on the modulation of the immune response in horses vaccinated with recombinant Clostridium tetani toxin. Twenty horses were vaccinated twice, with an interval of 21 days between doses, and equally divided into two groups: the first group was supplemented orally for 42 days with feed containing viable spores of B. toyonensis (1 × 108) mixed with molasses (40 ml), starting 7 days before the first vaccination; the second (control) group received only feed mixed with molasses, starting 7 days before the first vaccination. Serum samples were collected to evaluate the humoral immune response using an in-house indirect enzyme-linked immunosorbent assay (ELISA), and peripheral blood mononuclear cells (PBMCs) were collected to evaluate cytokine transcription (qPCR). For the specific IgG-anti-rTENT titer, the supplemented group had ELISA values that were four times higher than those of the control group (p < 0.05). The supplemented group also showed higher ELISA values for the IgGa and IgGT sub-isotypes compared to the control group. In PBMCs stimulated with B. toyonensis, relative cytokine transcription of the supplemented group showed 15-, 8-, 7-, and 6-fold increases for IL1, TNFα, IL10 and IL4, respectively. When stimulated with a vaccine antigen, the supplemented group showed 1.6-, 1.8-, and 0.5-fold increases in IL1, TNFα, and IL4, respectively, compared to the control group. Horses supplemented with B. toyonensis had a significantly improved vaccine immune response compared to those in the control group, which suggests a promising approach for improving vaccine efficacy with probiotics.

Asunto(s)

Bacillus , Enfermedades de los Caballos , Probióticos , Animales , Caballos/inmunología , Bacillus/inmunología , Probióticos/administración & dosificación , Probióticos/farmacología , Enfermedades de los Caballos/prevención & control , Enfermedades de los Caballos/inmunología , Enfermedades de los Caballos/microbiología , Tétanos/prevención & control , Tétanos/inmunología , Toxoide Tetánico/inmunología , Toxoide Tetánico/administración & dosificación , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/administración & dosificación , Masculino , Alimentación Animal , Femenino , Dieta/veterinaria , Citocinas/metabolismo

RESUMEN

Leptospirosis is an infectious disease with a worldwide distribution, which represents a major challenge in animal production across developing countries, mainly in tropical areas. Horses are particularly susceptible to the disease, presenting manifestations ranging from subclinical to the development of uveitis that compromises the visual health of the animals. In recent years, serological studies have been carried out in equid populations from America, demonstrating high exposure. For this reason, the aim of this study was to demonstrate microbiologically and molecularly the presence of the members of the genus Leptospira in urine samples from equids in an endemic state of leptospirosis in Mexico, and to detect the serological presence of anti-Leptospira antibodies in the sampled animals. For this reason, blood and urine samples were collected from 28 horses and one mule from three localities in the state of Veracruz, Mexico. Urine samples were inoculated in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium, and the recovered isolates were typed using a short Multi Locus Sequence Typing scheme. Amplifications of the expected size were subjected to sequencing, and the recovered sequences were compared with those of reference deposited in GenBank using the BLAST tool. To identify their phylogenetic position, we performed a phylogenetic reconstruction using the maximum likelihood method. Additionally, Microscopic Agglutination test was performed on the serum samples to identify anti-Leptospira antibodies. We recovered 16 urine isolates which tested positive for the presence of Leptospira DNA. The phylogenetic reconstruction and the MLST analysis confirmed the presence of several genotypes of Leptospira interrogans and Leptospira santarosai. An overall serological frequency of 97.1 % was detected. Our results represent the first record of the presence of Leptospira through bacteriological isolates in equids from Mexico.

Asunto(s)

Anticuerpos Antibacterianos , Enfermedades de los Caballos , Leptospira , Leptospirosis , Filogenia , Animales , Leptospirosis/veterinaria , Leptospirosis/microbiología , Leptospirosis/epidemiología , México/epidemiología , Caballos/microbiología , Enfermedades de los Caballos/microbiología , Leptospira/aislamiento & purificación , Leptospira/genética , Leptospira/clasificación , Anticuerpos Antibacterianos/sangre , Leptospira interrogans/aislamiento & purificación , Leptospira interrogans/genética , Leptospira interrogans/clasificación , Tipificación de Secuencias Multilocus , Análisis de Secuencia de ADN , ADN Bacteriano/genética

RESUMEN

The reproductive features of equine leptospirosis are often neglected. Equine genital leptospirosis is characterized as a silent chronic syndrome, and besides abortions, leads to placental abnormalities, stillbirths, and birth of weak foals. This study aimed to study the occurrence of placental abnormalities associated with Leptospira interrogans infection in naturally infected mares under field conditions. The studied herd had a high occurrence of placentitis and abortions. Ten pregnant mares, eight with placental abnormalities on ultrasonography and were selected. Serum and cervicovaginal mucus (CVM) samples were collected for serology and PCR, respectively. Positive samples in lipL32-PCR were submitted to the sequencing of the secY gene. In lipL32-PCR of CVM, five out of 10 (50%) mares were positive and all were characterized as Leptospira interrogans. Our results highlight the presence of placental abnormalities in the reproductive subclinical leptospirosis syndrome. We encourage field veterinarians to include leptospirosis testing in their reproductive management.

Asunto(s)

Enfermedades de los Caballos , Leptospira interrogans , Leptospirosis , Enfermedades Placentarias , Placenta , Complicaciones Infecciosas del Embarazo , Animales , Caballos , Leptospirosis/veterinaria , Leptospirosis/microbiología , Leptospirosis/epidemiología , Leptospirosis/complicaciones , Enfermedades de los Caballos/microbiología , Embarazo , Femenino , Leptospira interrogans/aislamiento & purificación , Enfermedades Placentarias/microbiología , Enfermedades Placentarias/veterinaria , Enfermedades Placentarias/patología , Complicaciones Infecciosas del Embarazo/veterinaria , Complicaciones Infecciosas del Embarazo/microbiología , Placenta/microbiología , Placenta/patología

RESUMEN

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a mucosal commensal of the lower genital tract in horses and is the most isolated bacterium causing endometritis in mares. The aim of this study was to determine the molecular diversity of S. zooepidemicus obtained from endometritis in mares in Buenos Aires province, Argentina. Thirty isolates obtained from the uterus of mares in 2005 and 2017 were studied. The MLST scheme was applied to identify the Argentinian genotypes and the clonal relationships and patterns of evolutionary descent were identified using the eBURST algorithm - goeBURST. Twenty six different Sequence types (STs) were identified, being only 11 of them previously reported in horses and also, from several host species and tissues. The other 15 STs were reported in Argentinian reproductive strains of mares in our study for the first time. The genotypes obtained from uterus in Argentina were not evenly distributed when all the published S. zooepidemicus STs were analysed, thus, it was not possible to establish that the same lineage circulates in our equine population. The fact that the identified genotypes were also reported in other countries, diverse samples and host species suggest that there is not a host, and an anatomical niche adaptation. Finally, the isolation of the same genotype in the vagina/clitoris and the uterus of the same mare highlights the versatility of S. zooepidemicus and its role as an opportunistic pathogen.

Asunto(s)

Endometritis , Genotipo , Enfermedades de los Caballos , Infecciones Estreptocócicas , Animales , Caballos/microbiología , Enfermedades de los Caballos/microbiología , Femenino , Argentina , Endometritis/veterinaria , Endometritis/microbiología , Infecciones Estreptocócicas/veterinaria , Infecciones Estreptocócicas/microbiología , Variación Genética , Tipificación de Secuencias Multilocus/veterinaria , Útero/microbiología , Streptococcus/genética , Streptococcus/aislamiento & purificación , Streptococcus/clasificación , Streptococcus equi/genética , Streptococcus equi/aislamiento & purificación , Streptococcus equi/clasificación

RESUMEN

A clinical case of an adult horse with invasive, ulcerative, proliferative, pyogranulomatous disease of the skin (tumor) in the shoulder region is presented. The mass had a granulomatous and crater-shaped appearance, with serosanguinous discharge and the presence of fistulas with caseous material. The tumor was removed by surgery and sent to the laboratory for diagnosis. Histopathology was performed using Grocott-Gomori methenamine silver stain. The presence of necrotic material, fibrosis, infiltrated cells, and brown-colored hyphae, characteristic of members of the genus Pythium, were observed. To identify the infecting species, conventional PCRs for the amplification of the ITS-1 was carried out. Histopathological and PCR tests confirmed infection by a Pythium insidiosum strain closely associated with previous records from the US and Central America. Our report represents the first molecularly confirmed case of equine pythiosis in Mexico.

Asunto(s)

Enfermedades de los Caballos , Pitiosis , Pythium , Animales , Pitiosis/diagnóstico , Pitiosis/microbiología , Pitiosis/patología , Caballos , Pythium/aislamiento & purificación , Pythium/genética , Pythium/clasificación , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/diagnóstico , México , Reacción en Cadena de la Polimerasa , ADN Espaciador Ribosómico/genética , Masculino , Histocitoquímica , Piel/patología , Piel/microbiología , Piel/parasitología

RESUMEN

Endometritis, the inflammation of the endometrium, is the leading cause of subfertility in mares, and therefore responsible for major economic losses in the horse industry worldwide. It is generally treated with uterine lavages combined with ecbolic agents and local or systemic antibiotics. However, since antibiotic overuse has been associated with antimicrobial resistance in mares with persistent endometritis, new prevention and treatment alternatives are needed. One such alternative could be the use of probiotic lactic acid bacteria (LAB) isolated from the host. Thanks to their species specificity, resident microbiota may restore ecological equilibrium within the host, and therefore, help prevent infections and improve physiological functions. In the present study, 257 bacterial strains were isolated from 77 healthy mares, and 88.76% (n = 228) of them were phenotypically classified as LAB. Within this group, 65.79% were able to inhibit at least one strain from each of the genera that most commonly cause equine endometritis (Streptococcus equi subsp. zooepidemicus, Escherichia coli, and Staphylococcus spp.). Five strains (RCE11, RCE20, RCE91, RCE99, and RCE167) were selected on the basis of their beneficial properties: ability to autoaggregate and adhere to equine epithelial cells, high inhibition of and co-aggregation with all the bacteria isolated from clinical cases of endometritis evaluated, and negative co-inhibition between one another. All five were finally identified as Enterococcus spp., namely E. faecium (two strains), E. hirae (two strains), and E. gallinarum (one strain). Further studies will assess their safety and biotechnological potential for the design of a multi-strain probiotic formula to prevent equine endometritis.

Asunto(s)

Endometritis , Enfermedades de los Caballos , Probióticos , Animales , Caballos , Femenino , Endometritis/veterinaria , Endometritis/prevención & control , Endometritis/microbiología , Enfermedades de los Caballos/prevención & control , Enfermedades de los Caballos/microbiología , Probióticos/farmacología , Probióticos/administración & dosificación , Lactobacillales/aislamiento & purificación , Lactobacillales/fisiología , Genitales Femeninos/microbiología

RESUMEN

Pythiosis is a disease caused by the oomycete Pythium insidiosum, mainly reported in equines, dogs and humans and directly transmitted through contaminant zoospores in aquatic environments. We report the first outbreak of equine pythiosis in five equines. Wound samples were submitted for diagnostic testing including mycological culture and nested PCR. Treatment approaches consisted of conventional and alternative therapies. Microbiological analyses were performed using water samples from the riverbanks close to where the animals had grazed. All animals were positive for P. insidiosum cultures, and two animals responded successfully to alternative therapy (ozone therapy). After culture and molecular analysis of environmental samples, the presence of P. insidiosum in one section of the Tietê River was confirmed through a 99% sequence identity. Phylogenetic analyses using the cytochrome oxidase II gene showed that the animal isolates clustered in clade I and the environmental isolates clustered in clade III. Although the environmental and wound isolates belonged to different genetic clades, we concluded that the Tietê River is an important source of infection by P. insidiosum and that research concerning environmental isolation of P. insidiosum from rivers and lakes should be strongly facilitated in Brazil.

Asunto(s)

Brotes de Enfermedades , Enfermedades de los Caballos , Pitiosis , Pythium , Animales , Brasil/epidemiología , Brotes de Enfermedades/veterinaria , Perros , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/microbiología , Caballos , Filogenia , Pitiosis/epidemiología , Pythium/clasificación

RESUMEN

Dermatophilus congolensis is a bacterial pathogen mostly of ruminant livestock in the tropics/subtropics and certain temperate climate areas. It causes dermatophilosis, a skin disease that threatens food security by lowering animal productivity and compromising animal health and welfare. Since it is a prevalent infection in ruminants, dermatophilosis warrants more research. There is limited understanding of its pathogenicity, and as such, there is no registered vaccine against D. congolensis. To better understanding the genomics of D. congolensis, the primary aim of this work was to investigate this bacterium using whole-genome sequencing and bioinformatic analysis. D. congolensis is a high GC member of the Actinobacteria and encodes approximately 2527 genes. It has an open pan-genome, contains many potential virulence factors, secondary metabolites and encodes at least 23 housekeeping genes associated with antimicrobial susceptibility mechanisms and some isolates have an acquired antimicrobial resistance gene. Our isolates contain a single CRISPR array Cas type IE with classical 8 Cas genes. Although the isolates originate from the same geographical location there is some genomic diversity among them. In conclusion, we present the first detailed genomic study on D. congolensis, including the first observation of tet(Z), a tetracycline resistance-conferring gene.

Asunto(s)

Dermatophilus/efectos de los fármacos , Dermatophilus/genética , Actinobacteria/genética , Animales , Antibacterianos/farmacología , Bovinos , Enfermedades de los Bovinos/metabolismo , Biología Computacional/métodos , Dermatophilus/metabolismo , Genoma Bacteriano , Infecciones por Bacterias Grampositivas/genética , Infecciones por Bacterias Grampositivas/microbiología , Infecciones por Bacterias Grampositivas/veterinaria , Enfermedades de los Caballos/microbiología , Caballos , Resistencia a la Tetraciclina/genética , Secuenciación Completa del Genoma/métodos

RESUMEN

Brazilian spotted fever (BSF) is a common tick-borne disease caused by Rickettsia rickettsii. Horses are the primary hosts of the main vector, Amblyomma sculptum, and are considered efficient sentinels for circulation of Rickettsia. Therefore, the aim of this study was to detect antibodies reactive to R. rickettsii antigens in horses from a non-endemic area in the north-central region of Bahia state, Brazil. Blood samples and ticks were collected from 70 horses from the municipalities of Senhor do Bonfim, Antônio Gonçalves, Pindobaçu, and Campo Formoso. The sera obtained were tested by an indirect immunofluorescence assay to detect anti-Rickettsia antibodies. Overall, 5.7% (4/70) of the horses reacted to R. rickettsii antigens. Ticks were collected from 18.6% (13/70) of the horses and were identified as A. sculptum and Dermacentor nitens. Despite being a non-endemic area, seropositive animals were identified in our study, suggesting circulation of rickettsial agents in the region. This is the first serological survey of this agent in horses from the north-central region of Bahia, and further studies are needed to understand the epidemiology of BSF in these locations.

Asunto(s)

Enfermedades de los Caballos/diagnóstico , Caballos/microbiología , Rickettsia rickettsii , Fiebre Maculosa de las Montañas Rocosas/veterinaria , Garrapatas , Animales , Brasil/epidemiología , Técnica del Anticuerpo Fluorescente , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/microbiología , Fiebre Maculosa de las Montañas Rocosas/diagnóstico , Fiebre Maculosa de las Montañas Rocosas/epidemiología , Garrapatas/clasificación , Garrapatas/microbiología

RESUMEN

Bacteria of the genus Ehrlichia are transmitted by ticks and also are an important cause of infection in wild and domestic mammals. Infection with Ehrlichia spp. has been reported in horses, especially in the USA, Nicaragua and Brazil. In this study, we report the parasitism by Amblyomma sculptum, Rhipicephalus microplus and Dermacentor nitens ticks in horses from a ranch located in south Pantanal wetland. Molecular and serological analyzes to determine infection by Ehrlichia spp. in horses and their respective ticks were carried out. A total of 12 horses were submitted to blood collection to investigate antibodies by indirect immunofluorescence assay (IFA) using Ehrlichia canis crude antigens and to be tested by polymerase chain reaction (PCR) in order to amplify fragments of the 16S rRNA, dsb, groEL and sodB gene of Ehrlichia spp. A total of 164 tick specimens were removed from horses, stored in isopropanol and later identified as D. nitens, A. sculptum and R. microplus. DNA from ticks were extracted and subjected to the same PCR assays to detect Ehrlichia spp. Anti-Ehrlichia spp. antibodies were detected in five/12 (41.7 %) horses by IFA, with antibody titers ranging from 40 to 160. All horse DNA samples were negative for the 16S rRNA, dsb, groEL and sodB of Ehrlichia spp. One A. sculptum female was positive to all target genes of Ehrlichia. This tick was parasitizing an Ehrlichia-seropositive horse with antibody titer of 80. Nucleotide sequences of 16S rRNA, dsb, groEL and sodB genes showed close relationship with different strains of Ehrlichia detected in wild mammals, Amblyomma ticks and horses from Brazil and Argentina. Detection of anti-Ehrlichia sp. antibodies suggests that horses have been exposed to an ehrlichial agent in the Pantanal. Future studies on Ehrlichia infection should be carried out to better elucidate and to bring new information about equine ehrlichiosis, since these animals are important hosts of ticks in the Brazilian Pantanal wetlands.

Asunto(s)

Amblyomma/microbiología , Ehrlichia/aislamiento & purificación , Ehrlichiosis/veterinaria , Enfermedades de los Caballos/microbiología , Infestaciones por Garrapatas/veterinaria , Animales , Brasil , Ehrlichiosis/microbiología , Femenino , Enfermedades de los Caballos/parasitología , Caballos , Masculino , Ninfa/microbiología , Infestaciones por Garrapatas/parasitología

RESUMEN

Equine infectious disease outbreaks may have profound economic impact, resulting in losses of millions of dollars of revenue as a result of horse loss, quarantine, and cancelled events. Early and accurate diagnosis is essential to limit the spread of infectious diseases. However, laboratory detection of infectious agents, especially the simultaneous detection of multiple agents, can be challenging to the clinician and diagnostic laboratory. Next-generation sequencing (NGS), which allows millions of DNA templates to be sequenced simultaneously in a single reaction, is an ideal technology for comprehensive testing. We conducted a proof-of-concept study of targeted NGS to detect 62 common equine bacterial, viral, and parasitic pathogens in clinical samples. We designed 264 primers and constructed a bioinformatics tool for the detection of targeted pathogens. The designed primers were able to specifically detect the intended pathogens. Results of testing 27 clinical samples with our targeted NGS assay compared with results of routine tests (assessed as a group) yielded positive percent agreement of 81% and negative percent agreement of 83%, overall agreement of 81%, and kappa of 0.56 (moderate agreement). This moderate agreement was likely the result of low sensitivity of some primers. However, our NGS assay successfully detected multiple pathogens in the clinical samples, including some pathogens missed by routine techniques.

Asunto(s)

Secuenciación de Nucleótidos de Alto Rendimiento/veterinaria , Enfermedades de los Caballos/diagnóstico , Animales , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/virología , Caballos

RESUMEN

Spotted fever group (SFG) rickettsiae are obligatory intracellular bacteria that cause disease in humans and other animals. Ixodid ticks are the principal vectors of SFG rickettsiae. The present study aimed to determine the prevalence and species identity of SFG rickettsiae in ticks and horses from urban and rural areas of western Cuba using PCR assays. Tick samples, collected from 79 horses, consisted of 14 Amblyomma mixtum adults, 111 Dermacentor nitens adults and 19 pools of D. nitens nymphs (2-5 individuals/pool). The PCR results revealed the presence of Rickettsia spp. in 64% of the A. mixtum adults, 16% of the D. nitens adults, and 11% of the pooled samples of D. nitens nymphs. In contrast, Rickettsia spp. was not detected in any of the 200 horse blood samples included in this study. DNA sequence data of the rickettsial 17 kDa antigen gene showed that Rickettsia amblyommatis was present in A. mixtum; and Rickettsia felis in D. nitens. This is the first report of R. felis in D. nitens in Cuba. The present study extends our knowledge of the potential vector spectrum and distribution of SFG rickettsiae pathogens in western Cuba.

Asunto(s)

Caballos , Ixodidae/microbiología , Rickettsia , Rickettsiosis Exantemáticas/veterinaria , Amblyomma/microbiología , Animales , Vectores Arácnidos/microbiología , Cuba/epidemiología , ADN Bacteriano/genética , Dermacentor/microbiología , Enfermedades de los Caballos/microbiología , Caballos/microbiología , Caballos/parasitología , Ninfa/microbiología , Patología Molecular , Reacción en Cadena de la Polimerasa/veterinaria , Rickettsia/genética , Rickettsia/aislamiento & purificación , Rickettsiosis Exantemáticas/epidemiología , Rickettsiosis Exantemáticas/microbiología , Infestaciones por Garrapatas/veterinaria

RESUMEN

Abstract Brazilian spotted fever (BSF) is a common tick-borne disease caused by Rickettsia rickettsii. Horses are the primary hosts of the main vector, Amblyomma sculptum, and are considered efficient sentinels for circulation of Rickettsia. Therefore, the aim of this study was to detect antibodies reactive to R. rickettsii antigens in horses from a non-endemic area in the north-central region of Bahia state, Brazil. Blood samples and ticks were collected from 70 horses from the municipalities of Senhor do Bonfim, Antônio Gonçalves, Pindobaçu, and Campo Formoso. The sera obtained were tested by an indirect immunofluorescence assay to detect anti-Rickettsia antibodies. Overall, 5.7% (4/70) of the horses reacted to R. rickettsii antigens. Ticks were collected from 18.6% (13/70) of the horses and were identified as A. sculptum and Dermacentor nitens. Despite being a non-endemic area, seropositive animals were identified in our study, suggesting circulation of rickettsial agents in the region. This is the first serological survey of this agent in horses from the north-central region of Bahia, and further studies are needed to understand the epidemiology of BSF in these locations.

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Resumo A Febre Maculosa Brasileira (FMB) é uma importante zoonose causada por Rickettsia rickettsii, sendo seu vetor o Amblyomma sculptum. Cavalos são os principais hospedeiros desse carrapato e considerados sentinelas eficientes para detectar a circulação de Rickettsia. O objetivo deste estudo foi detectar anticorpos reativos a antígenos de R. rickettsii em cavalos de uma área não-endêmica na região Centro-Norte da Bahia, Brasil. Foram coletadas amostras de sangue e carrapatos de 70 cavalos nos municípios de Senhor do Bonfim, Antônio Gonçalves, Pindobaçu e Campo Formoso. Os soros obtidos foram testados pela reação de imunofluorescência indireta para detectar anticorpos anti-Rickettsia. No geral, 5,7% (4/70) dos cavalos reagiram sorologicamente para antígenos de R. rickettsii. Os carrapatos foram coletados em 18,6% (13/70) dos cavalos e foram identificados como A. sculptum e Dermacentor nitens. Embora seja uma área não endêmica, foram observados animais soropositivos, sugerindo a circulação de riquétsias na região estudada. Este é o primeiro estudo de levantamento sorológico desse agente em cavalos do Centro-Norte da Bahia. São necessários mais estudos para compreender a epidemiologia da FMB nesses locais.

Asunto(s)

Animales , Rickettsia rickettsii , Garrapatas/clasificación , Garrapatas/microbiología , Fiebre Maculosa de las Montañas Rocosas/veterinaria , Enfermedades de los Caballos/diagnóstico , Caballos/microbiología , Brasil/epidemiología , Fiebre Maculosa de las Montañas Rocosas/diagnóstico , Fiebre Maculosa de las Montañas Rocosas/epidemiología , Técnica del Anticuerpo Fluorescente , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/epidemiología

RESUMEN

Leptospirosis in horses is often associated with reproductive disorders. In the southern states of Brazil, horses are used for various jobs and cultural practices; nevertheless, serological surveillance for Leptospira is rare. Therefore, the objective of this study was to determine the seroprevalence of Leptospira spp. in horses in southern Brazil, as well as to identify the risk factors for infection and its impacts on reproduction. We performed microscopic agglutination tests for 12 serovars that corresponding 9 serogroup (Sejroe, Icterohaemorrhagiae, Australis, Pyrogenes, Pomona, Canicola, Grippotyphosa, Tarassovi and Ballum) in 595 samples from 60 herds. A brief history was obtained to analyze risk factors for reproductive disorders. A total of 45.9% of the tested horses were seropositive, of which the most frequent serogroups were Icterohaemorrhagiae (Icterohaemorrhagiae and Copenhageni serovars) and Ballum (Ballum serovar). Simple infections were found in 45.4% of seropositive animals, while mixed infections occurred in 54.6% of horses. There was a correlation between seropositivity and age and sex, that is, seropositivity was more frequent in animals over 6 years old and in females. There was no correlation between seropositivity and reproductive disorders. We conclude that there is a high seroprevalence of Leptospira spp. in southern Brazil with predominance of Icterohaemorrhagiae serogroup, mainly in older animals. Location, breeds, contact with dogs or other domestic animals are not risk factors, whereas gender is a risk factor. Reproductive disorders are not due to leptospirosis in the study region.

Asunto(s)

Enfermedades de los Caballos/epidemiología , Leptospira/inmunología , Leptospirosis/veterinaria , Reproducción , Factores de Edad , Pruebas de Aglutinación/veterinaria , Animales , Anticuerpos Antibacterianos/sangre , Brasil/epidemiología , Perros , Femenino , Enfermedades de los Caballos/microbiología , Enfermedades de los Caballos/fisiopatología , Caballos , Leptospirosis/epidemiología , Leptospirosis/fisiopatología , Masculino , Embarazo , Complicaciones Infecciosas del Embarazo/epidemiología , Complicaciones Infecciosas del Embarazo/microbiología , Complicaciones Infecciosas del Embarazo/fisiopatología , Complicaciones Infecciosas del Embarazo/veterinaria , Prevalencia , Factores de Riesgo , Rumiantes , Estudios Seroepidemiológicos

RESUMEN

We document the causes and pathological findings in 50 cases of equine pneumonia and pleuritis in Southern Brazil. Suppurative (17/50), pyogranulomatous (14/50), aspiration (5/50), mycotic (4/50), bronchointerstitial (3/50), embolic (3/50) and eosinophilic granulomatous pneumonia (1/50) and pleuritis (3/50) were the main conditions identified. Streptococcus spp. were identified in 11 cases of suppurative pneumonia. Suppurative pneumonia was further divided into acute (8/17), subacute (6/17) and chronic (3/17) based on the morphological pattern of lesions. Rhodococcus equi was identified in all cases of pyogranulomatous pneumonia and exclusively affected young foals.

Asunto(s)

Enfermedades de los Caballos/microbiología , Pleuresia/veterinaria , Neumonía/veterinaria , Animales , Brasil , Caballos

RESUMEN

Detection of Leptospira interrogans is difficult as a result of intermittent leptospiruria and brief leptospiremia. Hence, diagnosis relies heavily on serologic testing, the reference method of which is the microscopic agglutination test (MAT). In horses, clinical leptospirosis has been associated with abortion, recurrent uveitis, and sporadic cases of hepatic and renal disease. Little information exists on the seroprevalence of antibodies to L. interrogans in equids in the United States; past nationwide studies suggest that the seroprevalence in some areas is as high as 77% (reciprocal titer ≥ 100). We tested sera from 124 apparently healthy horses previously submitted for equine infectious anemia (EIA) serology using MAT for 6 serovars-Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, and Pomona. When using a reciprocal MAT titer cutoff of ≥ 100, 102 of 124 (82%) of the samples were positive for at least one serovar. Seropositivity was significantly associated with increasing age. Query of specimens from clinical cases submitted to the Colorado State University Veterinary Diagnostic Laboratory for MAT since 2010 indicated significantly greater seroprevalence (p = 0.015) of pathogenic serovar Pomona in clinical cases compared to sera submitted from healthy equids for routine EIA testing. Information from our diagnostic laboratory submission forms also suggests a correlation between uveitis or other ophthalmic problems and serovar Pomona.

Asunto(s)

Enfermedades de los Caballos/epidemiología , Leptospirosis/veterinaria , Factores de Edad , Animales , Anticuerpos Antibacterianos/sangre , Colorado/epidemiología , Femenino , Enfermedades de los Caballos/microbiología , Caballos , Leptospira , Leptospirosis/epidemiología , Leptospirosis/microbiología , Masculino , Prevalencia , Estudios Seroepidemiológicos , Serogrupo

RESUMEN

Considering the lack of studies evaluating the performance of commercially available methods for diagnosis of Clostridioides (Clostridium) difficile infection (CDI) in animals, the present study aimed to assess an immunochromatographic test for detection of glutamate dehydrogenase (GDH) and A/B toxins of C. difficile, also evaluated by an ELISA kit, in foals and neonatal piglets. Intestinal contents of 47 piglets and feces of 35 foals were tested to GDH antigen and A/B toxins in a lateral flow method (Ecodiagnostica, Brazil). Also, these samples were submitted to A/B toxin detection by an ELISA kit (C. difficile Tox A/B II, Techlab Inc., USA), using the toxigenic culture (TC) as the reference method. The GDH component of the lateral flow test showed sensitivity and negative predictive value (NPV) of 100% and a high specificity in samples of piglets (82.61%) and foals (100%). Detection of A/B toxins using the lateral flow test and the ELISA resulted in a specificity of 100% in samples of both species. On the other hand, the sensibility ranged from 54.2 to 90% for the ELISA and from 12.5 to 60% for the lateral flow test for piglets' and foals' samples, respectively. In conclusion, the present work suggests that the lateral flow test for GDH detection could be a useful method for diagnosing CDI in these species. On the other hand, the low sensitivity of the lateral flow test for A/B toxins might compromise its utility in piglets.

Asunto(s)

Toxinas Bacterianas/análisis , Clostridioides difficile/aislamiento & purificación , Diarrea/veterinaria , Ensayo de Inmunoadsorción Enzimática/métodos , Glutamato Deshidrogenasa/análisis , Enfermedades de los Caballos/microbiología , Inmunoensayo/métodos , Enfermedades de los Porcinos/microbiología , Animales , Animales Recién Nacidos/microbiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/metabolismo , Brasil , Clostridioides difficile/enzimología , Clostridioides difficile/metabolismo , Infecciones por Clostridium/diagnóstico , Infecciones por Clostridium/microbiología , Infecciones por Clostridium/veterinaria , Diarrea/diagnóstico , Diarrea/microbiología , Heces/microbiología , Glutamato Deshidrogenasa/metabolismo , Enfermedades de los Caballos/diagnóstico , Caballos , Inmunoensayo/veterinaria , Porcinos , Enfermedades de los Porcinos/diagnóstico