Assessing the impact of various tuberculin PPD brands on bovine tuberculosis diagnosis.

Although several brands of tuberculin purified protein derivatives (PPDs) are available for diagnosing bovine tuberculosis (bTB), comparative studies to determine their diagnostic accuracy are infrequent. In Ecuador we compared two different PPD brands for bTB diagnosis using skin testing and measuring skin thickness increase. Additionally, we evaluated four PPD brands, including those used for skin testing, in the Bovine Tuberculosis Interferon Gamma Test (IFN-γ test) measuring IFN-γ induction in whole blood. The study included 17 naturally tuberculosis-infected PPD and IFN-γ test positive bovines. Both the field and laboratory results showed significant differences in classifying the 17 bovines as bTB positive or negative. We hypothesize that several factors, such as the genetic background of the cows, sensitization to environmental mycobacteria, M. bovis strains involved in the bTB infection, and the manufacturing procedures of the PPDs, could have influenced the immune reaction toward the different tuberculin PPD brands. Our study emphasizes the necessity for comparative studies aimed at determining the diagnostic accuracy of PPD brands for bTB diagnosis as well as the development of standardized methods for PPD production and potency determination.

First report and molecular identification of Trypanosoma (Duttonella) vivax outbreak in cattle population from Ecuador.

The bovine trypanosomosis is responsible for economic losses from tropical and subtropical areas of Africa and Latin America. This disease is characterized by fever, anaemia, loss of production and even death. Few studies have been carried out in Ecuador regarding Trypanosoma spp. presence but the species has not been determined in cattle and those have only determined the presence of genus, but not the species. The aim of this study was to identify and characterize the trypanosome species involved in the suspected bovine trypanosomosis outbreak reported in Convento Village in Manabí Province located in the coastal region of Ecuador. Twenty cattle from three farms were sampled. Three samples were positive for T. vivax, using an end-point polymerase chain reaction (PCR) to amplify a fragment of the cathepsin L-like cysteine protease (CatL-like) gene. A phylogenetic tree analysis of these three Ecuadorian isolates showed a close relationship with isolates from South America (Colombia, Brazil and Venezuela) and West Africa (Nigeria). This is the first report of T. vivax in Ecuadorian cattle.

Molecular characterization of Eimeria sp. from Galápagos giant tortoises (Chelonoidis spp.).

Galápagos giant tortoises are an essential component of their ecosystem and evaluation of parasites in their populations is essential for the management of conservation processes. Coccidiosis is the most common intestinal infection in free-living and captive reptiles. The aim of this study was to characterize molecularly the presence of Eimeria sp. in captive reared giant tortoises from Santa Cruz, Santiago, Española, and Pinzon Islands hatched and housed at the tortoise rearing center on Santa Cruz Island, Galápagos, by sequencing of the 18S rRNA gene. Galápagos. All samples were previously analyzed by coproparasitoscopic flotation technique and PCR for molecular identification. The results obtained by microscopy examination showed oocysts in all samples. PCR and sequencing indicated the presence Eimeria sp., showing a similarity percentage of 98% with Eimeria environmental. In conclusion, we identified a group of coccidia of the genus Eimeria sp. (MK909931) in Galápagos tortoises.

Prevalence of bovine tuberculosis in slaughtered cattle identified by nested-PCR in abattoirs from two dairy areas of Ecuador.

Bovine tuberculosis (bTB) is a chronic granulomatous disease that primarily affects lung tissue and lymph nodes (LN) in cattle, with economic impact on their productivity. Furthermore, it is potential zoonoses that may cause public health hazard. In this study, we evaluated the presence of bTB in two abattoirs: Cayambe and Pelileo countries located in the Ecuadorian provinces of Pichincha and Tungurahua, respectively. In total, 578 cattle were sampled (Cayambe 271 and Pelileo 307): 1,156 LN and 578 lung tissue samples were collected to apply in vitro culture and nested-PCR, respectively. The results determined a total apparent prevalence of 4.33%, with 4.06% at Cayambe's abattoir and 4.56% at Pelileo's abattoir. Additionally, the Bayesian analysis showed a total true prevalence of 2.51%, with 89.7% of sensitivity and 97.6% of specificity. The risk factors were evaluated by the use of simple logistic regressions with and without the random effect of places of origin. Associations of the origin of cattle in the selected slaughterhouses were found. The results showed an efficient method for the detection of bTB, which could identify a large number of infected animals, and the usefulness of lung tissue samples for early diagnosis of the disease was demonstrated in this study.

Situation of bovine tuberculosis in Ecuador.

Bovine tuberculosis (BTB) is a chronic and contagious disease that affects domestic animals, wildlife, and humans. Caused by Mycobacterium bovis, BTB causes major economic losses and poses a serious constraint to international livestock trade. Moreover, in developing countries where BTB controls are lacking, M. bovis is a public health concern. In most developing countries, the prevalence of BTB in livestock is unknown because the information is either not reported or not available. In Ecuador, there is no national BTB control program. This article reviews the BTB situation in Ecuador by examining exhaustive data from tuberculin testing surveys and slaughterhouse surveillance studies conducted in 1972-2008 in a variety of the country's geographic areas. In Ecuador, several factors, including the dairy industry's expansion (preempted by the high demand for milk and its by-products), intensified efforts to increase the cattle population, the presence of M. bovis, and a lack of BTB controls, have caused a rise in BTB prevalence, and consequently, a growing push for the implementation of a national BTB control program.

Situation of bovine tuberculosis in Ecuador / Situación de la tuberculosis bovina en el Ecuador

Bovine tuberculosis (BTB) is a chronic and contagious disease that affects domestic animals, wildlife, and humans. Caused by Mycobacterium bovis, BTB causes major economic losses and poses a serious constraint to international livestock trade. Moreover, in developing countries where BTB controls are lacking, M. bovis is a public health concern. In most developing countries, the prevalence of BTB in livestock is unknown because the information is either not reported or not available. In Ecuador, there is no national BTB control program. This article reviews the BTB situation in Ecuador by examining exhaustive data from tuberculin testing surveys and slaughterhouse surveillance studies conducted in 1972-2008 in a variety of the country's geographic areas. In Ecuador, several factors, including the dairy industry's expansion (preempted by the high demand for milk and its by-products), intensified efforts to increase the cattle population, the presence of M. bovis, and a lack of BTB controls, have caused a rise in BTB prevalence, and consequently, a growing push for the implementation of a national BTB control program.

La tuberculosis bovina es una enfermedad contagiosa crónica que afecta a los animales domésticos, los animales salvajes y los seres humanos. Es producida por Mycobacterium bovis; causa grandes pérdidas económicas y plantea una grave limitación para el comercio ganadero internacional. Por otro lado, en los países en desarrollo donde no hay controles de la tuberculosis bovina, la infección por M. bovis representa un problema de salud pública. En la mayoría de los países en desarrollo, la prevalencia de tuberculosis en el ganado se desconoce porque la información no se comunica o no se consigue. En el Ecuador no hay un programa nacional de control de la tuberculosis bovina. En este artículo se revisa la situación de la tuberculosis bovina en el Ecuador, sobre la base de un análisis de los datos exhaustivos obtenidos de encuestas sobre pruebas de tuberculina y de los estudios de vigilancia llevados a cabo en mataderos entre 1972 y 2008 en varias zonas geográficas del país. En el Ecuador varios factores, como el crecimiento de la industria láctea (impulsado por la alta demanda de leche y sus derivados), los intensos esfuerzos para aumentar la población bovina, la presencia de M. bovis y la falta de controles de la tuberculosis bovina, han ocasionado un aumento de la prevalencia de esta y, en consecuencia, representan una motivación creciente para llevar a cabo un programa nacional de control de la tuberculosis bovina.

Post-mortem examination and laboratory-based analysis for the diagnosis of bovine tuberculosis among dairy cattle in Ecuador.

Veterinary inspection in slaughterhouses allows for the detection of macroscopic lesions reminiscent of bovine tuberculosis, but the presence of Mycobacterium bovis must be confirmed by laboratory methods. This study aimed at comparing the performances of the standard diagnostic tools used to identify M. bovis in tissue specimens sampled from suspicious animals. During a two years period, 1390 cattle were inspected at the Machachi abattoir in the Mejia canton - Ecuador. A total of 33 animals with granulomatous lesions were detected, representing 2.33% (16/687) and 2.42% (17/703) animals examined in 2007 and 2008, respectively. Ninety-four tissue specimens were sampled and screened for the presence of mycobacteria. Acid-fast bacilli were identified in one third of the suspicious cattle (11/33) and suggestive microscopic lesions in 27.3% (9/33) of the samples examined by direct microscopy and histopathology, respectively. Culturing on Stonebrink medium and 16S-rRNA-based polymerase chain reaction (PCR) yielded 36.4% (12/33) and 27.3% (9/33) of positives, respectively. Compared to culture, other diagnostic procedures displayed a lower sensitivity, with 56.5% for PCR, and 43.5% for direct microscopy and histopathology; however, the specificity was higher (94.4% for PCR and microscopy, and 97.2% for histopathology). We conclude that reliable post-mortem laboratory testing either requires the combination of a set of available diagnostic tools or necessitates the development of improved new-generation tools with better sensitivity and specificity characteristics.

European 1: a globally important clonal complex of Mycobacterium bovis.

We have identified a globally important clonal complex of Mycobacterium bovis by deletion analysis of over one thousand strains from over 30 countries. We initially show that over 99% of the strains of M. bovis, the cause of bovine tuberculosis, isolated from cattle in the Republic of Ireland and the UK are closely related and are members of a single clonal complex marked by the deletion of chromosomal region RDEu1 and we named this clonal complex European 1 (Eu1). Eu1 strains were present at less than 14% of French, Portuguese and Spanish isolates of M. bovis but are rare in other mainland European countries and Iran. However, strains of the Eu1 clonal complex were found at high frequency in former trading partners of the UK (USA, South Africa, New Zealand, Australia and Canada). The Americas, with the exception of Brazil, are dominated by the Eu1 clonal complex which was at high frequency in Argentina, Chile, Ecuador and Mexico as well as North America. Eu1 was rare or absent in the African countries surveyed except South Africa. A small sample of strains from Taiwan were non-Eu1 but, surprisingly, isolates from Korea and Kazakhstan were members of the Eu1 clonal complex. The simplest explanation for much of the current distribution of the Eu1 clonal complex is that it was spread in infected cattle, such as Herefords, from the UK to former trading partners, although there is evidence of secondary dispersion since. This is the first identification of a globally dispersed clonal complex M. bovis and indicates that much of the current global distribution of this important veterinary pathogen has resulted from relatively recent International trade in cattle.

Comparative intradermal tuberculin test in dairy cattle in the north of Ecuador and risk factors associated with bovine tuberculosis.

We studied the prevalence of bovine tuberculosis (BTB) in Mejia canton, the major dairy cattle production area in northern Ecuador. Twenty dairy herds comprising 2,022 cattle were selected. In 2007, each animal was tested using the comparative intradermal tuberculin test (CITT). In 2008, a follow-up test was performed in the same herds. The true annual incidence was 1.70%, and the true prevalence was 7.41% and 7.13% in 2007 and 2008, respectively. The prevalence was 0.27% and 0.57% in medium-sized herds in 2007 and 2008, respectively, compared with 8.63% and 8.43% in large herds (P < 0.01). The number of skin test-positive cases also increased significantly with age (P = 0.03), contacts with other species of animals (P < 0.01), and introduction of new cattle (P = 0.04). Herd prevalence was 55% in 2007 and 65% in 2008. This study shows the lack of knowledge in cattle farmers about this zoonosis and the necessity for a national BTB control program in Ecuador.

Preliminary observations on Mycobacterium spp. in dairy cattle in Ecuador.

This study evaluated bovine tuberculosis in Mejia canton, a major dairy cattle production region in Ecuador. Randomly selected cattle (1,012 from 59 farms) classified according to herd size were tested by the single tuberculin test (STT). Sixty days later, positive reactors were tested again by the comparative tuberculin test (CTT). In addition, tissue samples from two STT-CTT-positive reactors detected on a farm were obtained in a local slaughterhouse and analyzed bacteriologically. A total of 4.24% of the cattle were positive in the STT and 3.85% were positive in the CTT, with the highest number (7.95%) in large herds versus 3.4% in medium herds and 0.3% in small herds. Mycobacterium bovis was isolated from mesenteric lymph nodes and lungs of one animal. A 16S ribosomal RNA-based polymerase chain reaction confirmed culture results and differentiated mycobacteria other than M. tuberculosis. This study confirms the zoonotic importance of tuberculosis in Ecuadorian dairy cattle with herd size likely to be a crucial parameter in the prevalence of the disease. The implementation of a national control program is necessary and should be based on the detection of positive cattle by STT in combination with CTT.