Spatial-temporal risk factors in the occurrence of rabies in Mexico.

Rabies is a zoonotic disease that affects livestock worldwide. The distribution of rabies is highly correlated with the distribution of the vampire bat Desmodus rotundus, the main vector of the disease. In this study, climatic, topographic, livestock population, vampire distribution and urban and rural zones were used to estimate the risk for presentation of cases of rabies in Mexico by co- Kriging interpolation. The highest risk for the presentation of cases is in the endemic areas of the disease, i.e. the States of Yucatán, Chiapas, Campeche, Quintana Roo, Tabasco, Veracruz, San Luis Potosí, Nayarit and Baja California Sur. A transition zone for cases was identified across northern Mexico, involving the States of Sonora, Sinaloa, Chihuahua, and Durango. The variables topography, vampire distribution, bovine population and rural zones are the most important to explain the risk of cases in livestock. This study provides robust estimates of risk and spread of rabies based on geostatistical methods. The information presented should be useful for authorities responsible of public and animal health when they plan and establish strategies preventing the spread of rabies into rabies-free regions of México.

Vaccination Strategies in a Potential Use of the Vaccine against Bovine Tuberculosis in Infected Herds.

Bovine tuberculosis (bTB) is a disease of cattle that represents a risk to public health and causes severe economic losses to the livestock industry. Recently, one of the strategies recommended for reducing the prevalence of the disease in animals is the use of the BCG vaccine, alone or in combination with proteins. It has been shown that the vaccine elicits a strong immune response, downsizes the number of animals with visible lesions, and reduces the rate of infection as well as the bacillary count. This paper, based on scientific evidence, makes suggestions about some practical vaccination alternatives that can be used in infected herds to reduce bTB prevalence, considering BCG strains, vaccine doses, routes of application, and age of the animals. Our conclusion is that vaccination is a promising alternative to be included in current control programs in underdeveloped countries to reduce the disease burden.

Descriptive and Time-Series Analysis of Rabies in Different Animal Species in Mexico.

The spatio-temporal epidemiology of rabies has related the influence of environmental factors and anthropogenic changes on the movements of the hematophagous bat Desmodus rotundus. In Mexico, D. rotundus is the main transmitter of the rabies virus for different livestock species, modifying annually the fluctuation of the number of cases of rabies and its dissemination in subtropical areas and regions considered free of the disease. The purpose of this study was to perform a descriptive analysis of the distribution of cases of rabies in Mexico, and to perform a time-series analysis to evaluate stationarity and to predict the number of cases for the following year. A total of 3,469 cases were reported in the period of interest, of which the 89.1% occurred in cattle, 4.3% in horses, 1.5% in sheep, 0.6% in goats, 0.01% in pig, 3.1% in vampire bats, 0.3% in cervids, 0.2% in skunks, 0.1% in insectivorous bats, 0.1% in foxes, 0.1% in buffaloes, and 0.02% in coatis; 0.5% were not identified. The most frequent antigenic variants reported were AgV11, AgV5, and AgV3, associated with D. rotundus. The distribution of cases in bats correlates with the distribution of cases in domestic and wild animals; however, cases were observed in wild species in non-endemic areas of Mexico, like the State of Chihuahua. The additive model used in the time-series analysis showed a seasonal pattern with a peak of cases at the beginning of each year, from January to March. The model showed a good predicting value; the Pearson correlation coefficient R2 was 0.705. The highest probability for the occurrence of rabies cases in the different species estimated by Ordinary Kriging was in the coast of the Gulf of Mexico, involving the states of Tamaulipas, Veracruz, Tabasco, Chiapas, and Yucatan. This study confirms that rabies in domestic and wild species is endemic in tropical and subtropical areas-however, cases have been observed in new geographic areas-and provides useful information to support actions to stop the spread of the rabies virus or the reservoir, and for planning vaccination strategies considering time and place.

Spatiotemporal analysis of rabies in cattle in central Mexico.

Spatial epidemiology of bat-transmitted rabies in cattle has been limited to spatial distribution of cases, an approach that does not identify hidden patterns and the spread resulting in outbreaks in endemic and susceptible areas. Therefore, the purpose of this study was to determine the relationship between the three variables average annual maximum, annual minimum temperature and precipitation in the region on the one hand, and the spatial distribution of cases on the other, using geographic information systems and co-Kriging considering that these environmental variables condition the existence of the rabies vector Desmodus rotundus. A stationary behaviour between the primary and the secondary variables was verified by basic statistics and moving window statistics. The directions of greater and lesser spatial continuity were determined by experimental cross-semivariograms. It was found that the highest risk for bovine paralytic rabies occurs in areas known as La Huasteca Potosina and La Sierra Gorda that are characterized by a maximum temperature of 29.5 °C, a minimum temperature of 16.5 °C and precipitation of 1200 mm. A risk estimation map was obtained for the presence of rabies with a determination coefficient greater than 95%, and a correlation coefficient greater than 0.95. Our conclusion is that ordinary co- Kriging provides a better estimation of risk and spatial distribution of rabies than simple Kriging, making this the method recommended for risk estimation and regional distribution of rabies.

Molecular epidemiology of cattle tuberculosis in Mexico through whole-genome sequencing and spoligotyping.

Mycobacterium bovis infection in cattle persists in Mexico, posing a threat to human health. Control of bovine tuberculosis, through the National Program Against Bovine Tuberculosis, has led to the decrease of disease prevalence in most of the country, except for high dairy production regions. Genotyping of M. bovis has been performed mainly by spoligotyping and variable number tandem repeats (VNTR), but higher resolution power can be useful for a finer definition of the spread of the disease. Whole genome sequencing and spoligotyping was performed for a set of 322 M. bovis isolates from different sources in Mexico: Baja California, Coahuila, Estado de Mexico, Guanajuato, Hidalgo, Jalisco, Queretaro and Veracruz, from dairy and beef cattle, as well as humans. Twelve main genetic clades were obtained through WGS and genetic diversity analysis. A clear differentiation of the Baja California isolates was seen as they clustered together exclusively. However, isolates from the central states showed no specific clustering whatsoever. Although WGS proves to have higher resolving power than spoligotyping, and since there was concordance between WGS and spoligotyping results, we consider that the latter is still an efficient and practical method for monitoring bovine tuberculosis in developing countries, where resources for higher technology are scarce.

Molecular Relationship between Strains of M. bovis from Mexico and Those from Countries with Free Trade of Cattle with Mexico.

The purpose of this study was to identify relationships between spoligotypes of M. bovis from cattle in Mexico and those reported in countries with free trade of cattle with Mexico: Australia, Canada, New Zealand and the United States of America. Mexican spoligotypes were obtained from isolates collected from cattle in different parts of the country. Spoligotypes from Canada and New Zealand were obtained from different reports in the literature. Those from the United States were obtained from the database of the National Veterinary Services Laboratory in APHIS-USDA. In order to perform the analysis in a single data set, spoligotypes were all converted to binary data and classified according to www.mbovis.org or www.pasteur-guadeloupe.fr:8081. Epidemiologic information included country and species infected. From 3,198 isolates, 174 different spoligotypes were obtained, 95 were orphans. Ninety one percent of the isolates came from the Unites States (n = 1,609) and Mexico (n = 1,323). Spoligotype SB0265 is shared between Canada and the United States in cattle and wildlife. Six spoligotypes, SB0673, SB0121, SB0145, SB0971, SB0140 and SB1165, were frequent in cattle and wildlife in the United States and cattle in Mexico, suggesting wide exchange of strains. Spoligotype SB0669 was found only in Mexico. Spoligotype SB0140 was the most common in Australia and the sixth in the United States and Mexico. In a phylogenetic analysis, spoligotype SB0140 appears as the oldest spoligotype in the data set, suggesting this as the ancestral spoligotype for all spoligotypes in the five countries. Some spoligotypes are shared by animals and humans, corroborating the zoonotic importance of M. bovis.

Gray fox (Urocyon cinereoargenteus) parasite diversity in central Mexico.

Mexico has a long history of parasitological studies in communities of vertebrates. However, the mega diversity of the country makes fauna inventories an ongoing priority. Presently, there is little published on the parasite fauna of gray foxes (Urocyon cinereoargenteus Schereber, 1775) and this study provides new records of parasites for gray foxes in central Mexico. It is a continuation of a series of previous parasitological studies conducted with this carnivore in Mexico from 2003 to the present. A total of 24 foxes in the Parque Nacional El Cimatario (PANEC) were trapped, anaesthetized, and parasites recovered. The species found were Dirofilaria immitis, Ctenocephalides canis, C. felis, Euhoplopsillus glacialis affinis (first report for gray foxes in Mexico) Pulex simulants, and Ixodes sp. Three additional gray fox carcasses were necropsied and the parasites collected were adult nematodes Physaloptera praeputialis and Toxocara canis. The intensive study of the gray fox population selected for the 2013-2015 recent period allowed for a two-fold increase in the number of parasite species recorded for this carnivore since 2003 (nine to 18 parasite species), mainly recording parasitic arthropods, Dirofilaria immitis filariae and adult nematodes. The parasite species recorded are generalists that can survive in anthropic environments; which is characteristic of the present ecological scenario in central Mexico. The close proximity of the PANEC to the city of Santiago de Queretaro suggests possible parasite transmission between the foxes and domestic and feral dogs. Furthermore, packs of feral dogs in the PANEC might have altered habitat use by foxes, with possible impacts on transmission.

Genetic diversity based on MIRU-VNTR profile of isolates of Mycobacterium bovis from Mexican cattle.

Bovine tuberculosis (bTB) is a disease caused by Mycobacterium bovis (M. bovis), which affects cattle, animal species and humans. To determinate the genetic structure of strains of M. bovis in mexican cattle, 467 isolates obtained from 2009 to 2010 from different regions of Mexico with known spoligotype were included in the study. The isolates were genotyped by interspersed repeated mycobacterial units-variable number tandem repeats (MIRU-VNTR) obtaining 13 MIRU-VNTR groups. When combining MIRU-VNTR patterns with its spolygotypes, the Hunter genetic discrimination index (HGDI), we obtained 421 genetic patterns distributed in 17 groups. The HGDI for the total loci was 0.99. The locus that presented the higher HGDI was 2461 (0.857), while the locus with the lowest HGDI was 2686 (0.239). When we analyzed our results, using just 6 or 8 MIRU-VNTR we obtained an discriminatory power of 0.8499 and 0.8875 respectively indicating lower HGDI than 12 MIRU-VNTR locus.

Evaluación de la Patogenicidad y Capacidad Protectora de un Inmungeno Fresco Combinado de Babesia Bigemina y Babesia Bovis / Evaluación of pathogenicity and protección capability of a fresh combined immunogen of Babesia bigemina and Babesia bovis

El objetivo del trabajo fue evaluar la inocuidad y la capacidad inmunoprotectora de un inmunógeno combinado de Babesia bigemina y Babesia bovis a una dosis de 108 y 109, respectivamente, bajo un desafío controlado. En el primer experimento (inocuidad) se utilizaron 16 bovinos Holstein provenientes de una zona libre de garrapatas y negativos a anticuerpos contra Babesia spp, mediante la prueba de inmunofluorescencia indirecta (IFI), los cuales fueron inoculados, vía intramuscular, con 1 X 109 eritrocitos infectados (EI) con una cepa atenuada de B. bigemina y 108 EI de una clona irradiada de B. bovis, ambas derivadas de cultivo in vitro. Se observaron cambios en las constantes fisiológicas a partir del día 7 y hasta el día 14 posinoculación (PI) en los animales sin verse éstos físicamente afectados. El porcentaje de eritrocitos parasitados (PEP) fue <0.01 por ciento. El segundo experimento (inmunogenicidad), se realizó tres meses PI y consistió en el desafío de 8 de los animales del experimento 1 con cepas virulentas de ambas especies del protozoario a una dosis de 108 EI de cada una. Cuatro animales adicionales sirvieron como grupo testigo. El grupo inmunizado presentó ligera disminución en el volumen celular aglomerado (VCA) con temperatura rectal (TR) sin cambios y PEP de 0.06 y <0.01 para B. bigemina y B. bovis, respectivamente. El grupo testigo presentó TR mayor a 400C, disminución del VCA (29 por ciento) y el PEP fue de 0.5 para B. bigemina y 0.03 para B. bovis; estos animales requirieron tratamiento para evitar su muerte. Se concluye que el inmunógeno combinado de B. bigemina y B. bovis a dosis altas no provoca reacciones clínicas severas en animales susceptibles y proporciona una sólida protección al desafío controlado con cepas virulentas

Anaplasma marginale: diferentes grados de virulencia en dos aislados mexicanos / Anaplasma marginale: differences in the virulence of two mexican isolates

En el presente trabajo se evaluó la virulencia de los aislados MEX-15-099 (MEX15) y MEX-31-096 (MEX31) de Anaplasma marginale. Dos grupos de cuatro bovinos susceptibles, fueron inoculados con una dosis de 1 ï 106 eritrocitos infectados por bovino. La temperatura rectal, el volumen celular aglomerado (VCA), la rickettsemia y la presencia de anticuerpos específicos se registraron durante el periodo experimental de 65 días. Los animales del grupo MEX15 alcanzaron un máximo PEI entre 8.8 por ciento y 34.7 por ciento; en contraparte, en los animales del grupo MEX31 se observaron valores máximos en el PEI entre 0.1 por ciento y 16.3 por ciento. Sólo los bovinos del grupo MEX15 presentaron fiebre >40§C, coincidente con la amplificación de la rickettsia. Se registró una pérdida media del VCA de 68.2 ñ 8.9 por ciento en los animales inoculados con MEX15 y de 50.1 ñ 8.9 por ciento en los inoculados con MEX31, esta diferencia fue significativa entre los grupos (P<0.05). Excepto por un animal infectado con MEX31, todos desarrollaron valores de DO entre 0.355 y 0.829. Todos los animales infectados con MEX15 desarrollaron anaplasmosis clínica, mientras que sólo uno de los infectados lo fue con el aislado MEX31. Las observaciones anteriores muestran una mayor virulencia para el aislado MEX15.

Evaluación en Condiciones de Campo de la Vacuna inactivada de Anaplasma Marginale Denominada Plazvax / Evaluation in field conditions of inactivated vaccines of anaplasma marginale named Plazvax

Para determinar la efectividad de la vacuna Plazvax© contra la anaplasmosis bovina, se utilizaron 32 novillos susceptibles a la enfermedad. Un grupo de 24 novillos fue inmunizado contra Anaplasma marginale, inoculando subcutáneamente 1 ml (dos veces, 21 días aparte) de la vacuna. Ocho animales fungieron como testigos sin vacunar. Un mes después de la segunda inmunización, los animales fueron trasladados a una zona endémica de anaplasmosis para ser desafiados naturalmente. A partir del día ocho posintroducción al potrero (PIP) y hasta el día 105 se realizó un seguimiento que incluyó observación de manifestaciones clínicas, temperatura rectal (TR), hematocrito (HT) y parasitemia (PP). Tres bovinos murieron de babesiosis y los 29 animales restantes tuvieron seguimiento dos veces por semana a partir del día 42 PIP. El día 52 PIP se detectaron los primeros animales infectados con Anaplasma marginale (6, todos del grupo vacunado), y cuatro fueron tratados con oxitetraciclinas (días 52-64 PIP). A partir del día 84 PIP los animales testigo presentaron anaplasmosis severa con TR promedio de 40.50C, HT promedio de 13.2 por ciento y PP promedio de 9.4 por ciento hacia el día de tratamiento (días 87-95 PIP). Los animales vacunados presentaron los siguientes valores promedios de TR, 40.2ºC; HT, 16.3 por ciento; y PP 3.3 por ciento en el día de tratamiento (días 93-105 PIP). A pesar de existir diferencia en PP entre los animales vacunados y testigos, todos padecieron anaplasmosis clínica, requiriendo tratamiento. En conclusión, en su forma actual la vacuna no es recomendable para usarse en México; se sugiere el empleo o introducción en la vacuna Plazvax, de material antigénico de uno o más aislados de A. marginale provenientes de distintas regiones geográficas de México y realizar estudios sobre su capacidad inmunoprotectora en bovinos