Etiología de la Mastitis bovina en Zamora-Chinchipe, Ecuador

Background: Cattle mastitis is a global sanitary and economic problem that affects human and animal health. Several microorganisms involved in its etiology, such as Mycoplasma bovis, make prevention and control more difficult. Aim. To deepen the bacterial etiology of cattle mastitis in Zamora-Chinchipe, Ecuador, where collected milk was observed to have Mollicutes. Materials and methods: A number of 247 milking cows were studied in 2015-2016 in herds from Zamora-Chinchipe province, selected in areas with adequate ecological characteristics. The California Mastitis Test (CMT) was run on milk samples from each cow whereas Polymerase Chain Reaction (PCR) was performed to detect Mollicutes, and a bacteriological diagnostic was made to identify other pathogens. A comparison of proportions analysis was conducted to analyze the results of Mollicute presence, and CMT in the cows per cantons, and between the pathogens identified in the Mollicute-positive cows. Chi-square helped evaluate the relation between the Mollicute diagnostic results and CMT, and the pathogens identified. Logistic regression was useful to compare the prevalence ratio (PR). Results: The frequency of subclinical mastitis was found at high levels (81.4%) through CMT, and a frequency of 33.2% of Mycoplasma spp.-positive cows. In the bacteriological diagnostic, Streptococcus spp. (27.2%) and Staphylococcus aureus (22.1%) showed the highest frequencies (P< 0.05). S. aureus was detected with the highest frequency in the Mycoplasma spp.-positive cows (43.8%; P<0.05). Conclusions: The high infection caused by Mycoplasma spp. and S. aureus was significant, due to their potential repercussion on the clinical-epidemiological behavior of bovine mastitis in the province. Key words:   Bovine mastitis, Mollicutes, Mycoplasma, PCR, StaphylococcusAntecedentes: La mastitis bovina, problema sanitario y económico a nivel global, tiene implicaciones para la salud animal y humana. Numerosos microorganismos involucrados en su etiología complican su prevención y control, como Mycoplasma bovis. Objetivo. Profundizar en la etiología bacteriana de la mastitis bovina en Zamora-Chinchipe, Ecuador, en cuyos rebaños se constató previamente la presencia de Mollicutes en leche de tanque. Materiales y métodos: Durante 2015 y 2016 se muestrearon 247 vacas en ordeño de rebaños bovinos lecheros en la provincia Zamora-Chinchipe, seleccionados en los conglomerados definidos en el territorio según sus características ecológicas. Las muestras de leche de cada vaca se investigaron mediante Prueba California para Mastitis (CMT), y para el diagnóstico microbiológico de Mollicutes se realizó Reacción en Cadena de la Polimerasa (PCR), así como el diagnóstico bacteriológico para la identificación de otros patógenos. Mediante comparación de proporciones se analizaron los resultados del diagnóstico de Mollicutes y CMT en vacas por cantones y entre los patógenos identificados en las vacas positivas a Mollicutes. A través de Chi Cuadrado se evaluó la relación entre los resultados del diagnóstico de Mollicutes y CMT, y de los patógenos identificados, y por regresión logística se comparó su Razón de Prevalencia (RP). Resultados: Se encontró elevada frecuencia de mastitis subclínica (81.4%) mediante CMT y una frecuencia de 33,2 % de vacas positivas a Mycoplasma spp. En el diagnóstico bacteriológico Streptococcus spp. (27,2%) y Staphylococcus aureus (22,1%) tuvieron las mayores frecuencias (P< 0,05). S. aureus se detectó con la mayor frecuencia en vacas positivas a Mycoplasma spp. (43,8%; P<0,05). Conclusiones: Se destaca la importancia de la elevada coinfección de Mycoplasma spp. y S. aureus por su repercusión potencial en el comportamiento clínico-epidemiológico de la mastitis bovina en la provincia. Palabras clave: Mastitis Bovina, Mollicutes, Mycoplasma, PCR, Staphylococcu

Molecular diagnosis, prevalence and importance of Zoonotic Vector-Borne pathogens in Cuban shelter dogs- a preliminary study

The present study aimed to determine the prevalence of zoonotic vector-borne pathogens, including Anaplasma platys, Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Ehrlichia canis and Rickettsia spp. in shelter dogs from Cuba. Blood samples were collected from 100 shelter dogs and examined by molecular methods. Overall, 85 (85%; 95% CI: 77.88-92.12) dogs tested positive for at least one vector-borne pathogen using species-specific qPCR assays. Among the positive samples, E. canis was the most prevalent 62% (95% CI: 52.32-71.68), followed by A. platys 40% (95% CI: 30.23-49.77) and Rickettsia felis 27% (95% CI: 18.15-35.85), whereas 36% (95% CI: 26.43-45.57) showed co-infections. All samples were negative for A. phagocytophilum and B. burgdorferi s.l. The presence of 248 Rhipicephalus sanguineus ticks collected from the dogs was not statistically associated with the occurrence of infections. Thrombocytopenia was the most frequent haematological alteration found in PCR-positive dogs; it was statistically associated with the presence of E. canis, as well as co-infections (p < 0.05). The phylogenetic analyses of A. platys and E. canis based on 16S rRNA, groEL and gltA genes showed a low genetic diversity between Cuban strains. The present study demonstrates the high prevalence of vector-borne pathogens with zoonotic potential in shelter dogs from Cuba

Molecular Diagnosis, Prevalence and Importance of Zoonotic Vector-Borne Pathogens in Cuban Shelter Dogs : A Preliminary Study

The present study aimed to determine the prevalence of zoonotic vector-borne pathogens, including Anaplasma platys, Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Ehrlichia canis and Rickettsia spp. in shelter dogs from Cuba. Blood samples were collected from 100 shelter dogs and examined by molecular methods. Overall, 85 (85%; 95% CI: 77.88-92.12) dogs tested positive for at least one vector-borne pathogen using species-specific qPCR assays. Among the positive samples, E. canis was the most prevalent 62% (95% CI: 52.32-71.68), followed by A. platys 40% (95% CI: 30.23-49.77) and Rickettsia felis 27% (95% CI: 18.15-35.85), whereas 36% (95% CI: 26.43-45.57) showed co-infections. All samples were negative for A. phagocytophilum and B. burgdorferi s.l. The presence of 248 Rhipicephalus sanguineus ticks collected from the dogs was not statistically associated with the occurrence of infections. Thrombocytopenia was the most frequent haematological alteration found in PCR-positive dogs; it was statistically associated with the presence of E. canis, as well as co-infections (p &lt; 0.05). The phylogenetic analyses of A. platys and E. canis based on 16S rRNA, groEL and gltA genes showed a low genetic diversity between Cuban strains. The present study demonstrates the high prevalence of vector-borne pathogens with zoonotic potential in shelter dogs from Cuba

Molecular detection and characterization of Hepatozoon canis in stray dogs from Cuba

Canine hepatozoonosis caused by Hepatozoon canis is a worldwide distributed tick-borne disease of domestic and wild canids that is transmitted by ingestion of Rhipicephalus sanguineus sensu lato (s.l.) ticks. The present study was aimed to determine the prevalence of Hepatozoon infections in 80 stray dogs from Havana Province in Cuba, and to confirm the species identity and phylogenetic relationships of the causative agent. Samples were screened by microscopical examination of thin blood smears for the presence of Hepatozoon spp. gamonts and by genus-specific SYBR green-based real-time PCR assay targeting the 18S rRNA gene. Direct microscopy examination revealed Hepatozoon gamonts in the peripheral blood of 8 dogs (10.0%; 95% CI: 4.80-18.0%), while 38 animals (47.5%; 95% CI: 36.8-58.4%) were PCR-positive, including all microscopically positive dogs. Hence, the agreement between the two detection methods was 'poor' (κ = 0.20). Hematological parameters did not differ significantly between PCR-positive and PCR-negative dogs (p > 0.05). The DNA sequences of the 18S rRNA gene of the Hepatozoon spp. from Cuban dogs showed a nucleotide identity >99% with those of 18S rRNA sequences of Hepatozoon canis isolates from Czech Republic, Brazil and Spain. Phylogenetic analysis revealed that obtained sequences clustered within the Hepatozoon canis clade, different from the Hepatozoon felis or Hepatozoon americanum clades. The present study represents the first molecular characterization of Hepatozoon canis in stray dogs within Cuba