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La revisión se enfocó en presentar de manera concisa las aportaciones que investigadores del INIFAP, han desarrollado directamente o en colaboración con investigadores de otras instituciones sobre diferentes aspectos de las enfermedades que afectan a la ganadería bovina en México. Se describen investigaciones sobre enfermedades virales como la rabia y la diarrea viral bovina; bacterianas como la anaplasmosis, brucelosis, tuberculosis, paratuberculosis, leptospirosis y enfermedad respiratoria bovina; de las enfermedades parasitarias se incluye a la infestación por garrapatas y a la babesiosis. Se identifican posibles líneas de investigación que pueden coadyuvar a mitigar el impacto de las enfermedades en la producción. Se señalan aportes sobre el desarrollo o adaptación de técnicas diagnósticas de tipo serológico y molecular y se considera el diagnóstico de resistencia a los ixodicidas. Además, se indican parámetros epidemiológicos de las enfermedades y se refieren los biológicos generados que comprenden vacuna contra rabia, anaplasmosis y babesiosis; bacterina contra leptospirosis y una bacterina-toxoide contra neumonías. Asimismo, se comentan las evaluaciones del uso de BCG contra tuberculosis y una vacuna de nueva generación contra la brucelosis. En la revisión se concluye que la investigación del INIFAP en salud animal debe forzosamente tener como perspectiva las ciencias ómicas. Solo así se complementará el entendimiento de los mecanismos de las enfermedades, el desarrollo de nuevas técnicas diagnósticas y el diseño de vacunas efectivas y seguras. De modo que el gran reto será el involucramiento del área de salud animal al concepto de "Una Salud".

Domestic swine have been introduced by humans into a wide diversity of environments and have been bred in different production systems. This has resulted in an increased risk for the occurrence and spread of diseases. Although viromes of swine in intensive farms have been described, little is known about the virus communities in backyard production systems around the world. The aim of this study was to describe the viral diversity of 23 healthy domestic swine maintained in rural backyards in Morelos, Mexico, through collection and analysis of nasal and rectal samples. Next-generation sequencing was used to identify viruses that are present in swine. Through homology search and bioinformatic analysis of reads and their assemblies, we found that rural backyard swine have a high degree of viral diversity, different from those reported in intensive production systems or under experimental conditions. There was a higher frequency of bacteriophages and lower diversity of animal viruses than reported previously. In addition, sapoviruses, bocaparvoviruses, and mamastroviruses that had not been reported previously in our country were identified. These findings were correlated with the health status of animals, their social interactions, and the breeding/rearing environment (which differed from intensive systems), providing baseline information about viral communities in backyard swine.

El aislamiento en cultivo celular es el método principal para el diagnóstico del virus de la necrosis pancreática infecciosa (VNPI). Aunque es un método confiable, resulta costoso y conlleva tres semanas en proporcionar un resultado. Para disminuir el tiempo en el diagnóstico y aumentar la sensibilidad para la detección del VNPI, el objetivo del presente estudio fue establecer y evaluar el uso de una PCR anidada (PCRa) para un diagnóstico rápido del VNPI. Para ello, se diseñaron dos pares de iniciadores basados en secuencias mexicanas. El primer par (RT-PCR) amplificó un producto de 682 pb y el segundo par (PCRa) 229 pb del gen VP2. Posteriormente, se infectaron 70 crías de truchas arcoíris (Oncorhynchus mykiss) con la cepa virulenta MEX3-CSM-05 a una dosis de 1X105.8 DICC50%/0.02 ml. De cada organismo se colectó el riñón, el bazo, los sacos pilóricos, el hígado, el intestino y las branquias. Para evaluar las pruebas, se utilizaron 26 truchas adultas clínicamente sanas de granjas comerciales del Estado de México. La frecuencia de detección del VNPI mediante RT-PCR en las branquias fue de 87.1 %, en el hígado 61.4 %, en los sacos pilóricos 61.4 %, en el riñón 58.6%, en el intestino 35.7 % y en el bazo 32.9 % (P

A viromics study on the intestinal contents of migratory wild ducks of the genera Anas, Mareca, Spatula, and Oxyura during their winter stay in Mexico showed the presence of the virus family Alloherpesviridae. The genus Cyprinivirus is part of this family and includes cyprinid herpesvirus 3 (CyHV3). This is the etiological agent of the lethal disease known as koi herpesvirosis, which affects different strains of Common Carp Cyrprinus carpio. In this study, samples of the contents of 87 wild duck intestines were analyzed by endpoint PCR, of which 7 samples were positive for the amplification of the TK gene fragment corresponding to CyHV3. These results contribute to the knowledge about the spread of this virus to other species of aquatic animals in areas where fish and ducks coexist.

Bovine paralytic rabies (BPR) is a form of viral encephalitis that is of substantial economic importance throughout Latin America, where it poses a major zoonotic risk. Here, our objective was to utilize a laboratory protocol to determine the relative copy number of the rabies virus (RABV) genome in different bovine brain anatomical structures using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). qRT-PCR quantifies the specific number of gene copies present in a sample based on fluorescence emitted after amplification that is directly proportional to the amount of target nucleic acid present in the sample. This method is advantageous owing to its short duration, reduced risk of contamination, and potential to detect viral nucleic acids in different samples more easily compared to other techniques. The brains of six rabid animals were divided into six anatomical structures, namely the Ammon's horn, cerebellum, cortex, medulla, pons, and thalamus. All brains were identified as positive for RABV antigens based on a direct immunofluorescence test. The same anatomical structures from the brains of four RABV-negative bovines were also assessed. RNA was extracted from each structure and used for qRT-PCR. An assay was performed to determine the copy numbers of RABV genes using an in vitro transcribed nucleoprotein gene. The standard curve used to quantify viral RNA exhibited an efficiency of 100% and linearity of 0.99. Analysis revealed that the cortex, medulla, and thalamus were the ideal CNS portions for use in RABV detection, based on the observation that these structures possessed the highest levels of RABV. The test specificity was 100%. All samples were positive, no false positives were detected. This method can be used to detect RABV in samples that contain low levels of RABV during diagnosis of BPR.

The production of Nile tilapia ( Oreochromis niloticus ) has good technological development; however, today, it is still necessary to make it more efficient. One way to increase efficiency is to prevent disease and improve the food conversion factor. Since previous investigations of tilapia microbiota detected a high proportion of organisms belonging to the order Actinomycetes, this study was to isolate, identify, and describe the species of bacteria microbiota belonging to the cultured Nile tilapia. These were done with Nile tilapia grown in a warm sub-humid climate during spring and summer seasons. The biopsy of different organs was performed for bacteriological culture and 16S rRNA sequencing analysis. From the 180 tissue samples, 49 isolates of the order Actinomycetes were obtained, representing ten species from seven genera: Microbacterium , Brevibacterium , Cellulomonas , Corynebacterium , Kocuria , Actinomyces , and Micrococcus . In spring, Microbacterium dominated, accounting for 74% of the total population. In the summer, lower diversity was observed, with 39% represented by Microbacterium . 16S rRNA sequencing analysis enabled the classification of Actinomyces neuii and Microbacterium lacticum as Kocuria varians and Agromyces indicus ; the classification of Microbacterium imperiale as Rhodococcus and Micrococcus luteus was confirmed. No sequences of K. varians have been reported in fish. Microbacterium dextranoliticum showed high similarity to environmental samples. Here is the first study that analyzes the bacteria population in tilapia at the genetic level with an ecosystem approach, present in healthy cultured tilapia, indicating their beneficial associations with the host, making them candidates as probiotics, among other possible functions, applicable in tilapia cultivation.

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.

We collected 724 blood samples from dairy cattle from six Mexican states, and tested them for the presence of antibodies against BLV using a commercial ELISA test. Our study groups consisted of 32 samples: 12 asymptomatic cows, 12 cows with lymphocytosis and 8 samples of tumor tissue of the abomasum and heart of cattle with lymphoma. We designed three pairs of primers to amplify the complete BLV env gene, and obtained a fragment of 1548 nucleotides in length with the sequenced products. According to the phylogenetic tree we constructed to identify the viral genotype, 96.87 % of the sequences grouped into genotype 1, while a single sample from a cow with lymphocytosis (3.13 %) was associated with genotype 3 sequences. The similarity between the Mexican BLV sequences ranged from 0.985-1.00. In addition, the proportion of non-synonymous and synonymous mutations indicated negative selection. We did not identify any conserved residues in the viral protein sequences that could be related to BLV infection stage in cattle. Proviral quantification was performed using quantitative polymerase chain reaction, and we used Mood´s median test as statistical analysis. We found no significant association between proviral load and phase of infection. The sequences showed high similarity without any association between BLV surface glycoprotein and the different infection stages, nor differences in the proviral load. BLV genotype 1 was identified as prevalent in the studied samples, and for the first time in Mexico, we identified BLV genotype 3 in cattle.

The aim of this research was to detect rabies virus in peripheral tissues in captive vampires. Vampire bats were inoculated with 106 MICLD50 of homologous rabies virus. Bats displayed clinical signs of rabies beginning on d 8 until the 19th d post-inoculation (pi). Rabies virus antigens were found in the brain of all rabid bats. Viral RNA was detected in brain, salivary gland and tongue tissue by RT-PCR and nested PCR (nPCR). Viral genome was also detected in organs unrelated to the central nervous system. Rabies virus was not detected in saliva nor documented from any tissues without occurrence of viral antigens in the brain. Host humoral response was most pronounced via the induction of viral neutralizing antibodies (VNA) from d 8 to 20 pi, having a peak at d 14 with 0.9 IU. Antibody levels were variable, but tended to remain high after inoculation, showing significant differences to the negative control group (P=0.001). This research is one of the few recent studies focused upon Desmodus rotundus and contributes to the basic knowledge of rabies virus pathogenesis, which is required for an understanding of perpetuation in a major viral reservoir in Latin America.

Vampire bats are the only mammals known to feed exclusively on blood from other animals, often from domestic cattle. We tested the hypothesis that the adaptation of vampire bats to hematophagy would have resulted in shared viral communities among vampire bats and cattle, as a direct result of historic spillover events occurring due to hematophagy. We analyzed the presence of different viruses in sample populations of sympatric bat and prey populations and searched for shared viruses between taxa. A limited number of DNA viral groups were detected within each species. However, there was no evidence for a shared viral community among the vampire bat and cattle populations tested.

Adaptation to specialized diets often requires modifications at both genomic and microbiome levels. We applied a hologenomic approach to the common vampire bat (Desmodus rotundus), one of the only three obligate blood-feeding (sanguivorous) mammals, to study the evolution of its complex dietary adaptation. Specifically, we assembled its high-quality reference genome (scaffold N50 = 26.9 Mb, contig N50 = 36.6 kb) and gut metagenome, and compared them against those of insectivorous, frugivorous and carnivorous bats. Our analyses showed a particular common vampire bat genomic landscape regarding integrated viral elements, a dietary and phylogenetic influence on gut microbiome taxonomic and functional profiles, and that both genetic elements harbour key traits related to the nutritional (for example, vitamin and lipid shortage) and non-nutritional (for example, nitrogen waste and osmotic homeostasis) challenges of sanguivory. These findings highlight the value of a holistic study of both the host and its microbiota when attempting to decipher adaptations underlying radical dietary lifestyles.

Porcine reproductive and respiratory syndrome (PRRS) is an economically frustrating viral disease of pigs, characterized by severe reproductive failure in pregnant sows and respiratory disorders in piglets and growing pigs. Several research groups around the world have developed PRRSV vaccines. Some of these have been effective; however, owing to the complications that the syndrome presents and the viral evasion of the immune system, vaccines have not always been 100% effective. Biotechnological tools, such as the generation of plant-derived vaccines, offer alternatives to obtain more stable biologics, free of fermentation and cold chains. According to the literature, these vaccines are cost effective. In this document, we present some of the vaccines that have been developed against PRRSV, both traditional and new, and describe some alternatives developed in plants.

SUMMARYWest Nile virus (WNV) is a mosquito-borne neurotropic viral pathogen maintained in an enzootic cycle between mosquitoes (vectors) and birds (natural hosts) with equids, humans, and other vertebrates acting as dead-end hosts. WNV activity in Mexico has been reported in several domestic and wild fauna and in humans, and the virus has been isolated from birds, mosquitoes, and humans. However, no serological studies have been conducted in monkeys, and only two in a limited number of crocodiles (Crocodylus moreletii). Here we present data on the prevalence of neutralizing antibodies against WNV in 53 healthy wild monkeys (49 Ateles geoffroyi and four Alouatta pigra), and 80 semi-captive healthy crocodiles (60 C. acutus and 20 C. acutus–C. moreletti hybrids) sampled during 2012. None of the monkey sera neutralized WNV, whereas 55% of the crocodile sera presented neutralizing antibodies against WNV. These results can contribute to the design of surveillance programmes in Mexico.

Papillomas occur more frequently in cattle than other domestic animals. The causal agent of bovine papillomatosis is a virus that belongs to the family Papillomaviridae. In Tamaulipas, Mexico, the virus is considered a serious problem and has impeded the export of cattle to the United States, resulting in serious economic losses. Owing to the lack of information regarding the subtypes of papillomaviruses that infect cattle in Mexico, the aim of this study was to determine the subtypes in Tamaulipas. Fifty-two warts were analyzed with the use of polymerase chain reaction (PCR) involving primers that amplify the E7 gene of bovine papillomavirus (BPV). The PCR products were sequenced to differentiate the BPV-1 and BPV-2 subtypes. The sequencing quality was determined with the use of MEGA 6.0 software. Comparison of the Tamaulipas sequences with those of known BPV types by means of the MUSCLE algorithm showed that 53% of the former were BPV-1 and 47% were BPV-2. The distribution of the 2 subtypes in the cattle was homogeneous. This study demonstrated the presence of BPV-1 and BPV-2 in cattle from Tamaulipas and constitutes the first molecular characterization of papillomas in Mexico.

The fecal virome comprises a complex diversity of eukaryotic viruses, phages and viruses that infect the host. However, little is known about the intestinal community of viruses that is present in wild waterfowl, and the structure of this community in wild ducks has not yet been studied. The fecal virome compositions of six species of wild dabbling ducks and one species of wild diving duck were thus analyzed. Fecal samples were collected directly from the rectums of 60 ducks donated by hunters. DNA and RNA virus particles were purified and sequenced using the MiSeq Illumina platform. The reads obtained from the sequencing were analyzed and compared with sequences in the GenBank database. Viral-related sequences from the Herpesviridae, Alloherpesviridae, Adenoviridae, Retroviridae and Myoviridae viral families showed the highest overall abundances in the samples. The virome analysis identified viruses that had not been found in wild duck feces and revealed distinct virome profiles between different species and between samples of the same species. This study increases our understanding of viruses in wild ducks as possible viral reservoirs and provides a basis for further studying and monitoring the transmission of viruses from wild animals to humans and disease outbreaks in domestic animals.

Since 1970, aquaculture production has grown. In 2010, it had an annual average rate of 6.3% with 59.9 million tons of product and soon could exceed capture fisheries as a source of fishery products. However, the occurrence of viral diseases continues to be a significant limiting factor and its control is important for the development of this sector. In aquaculture farms, fish are reared under intensive culture conditions, and the use of viral vaccines has enabled an increase in production. Several types of vaccines and strategies of vaccination have been developed; however, this approach has not reached the expected goals in the most susceptible stage (fingerlings). Currently, there are inactivated and recombinant commercial vaccines, mainly for salmonids and cyprinids. In addition, updated genomic and proteomic technology has expedited the research and expansion of new vaccine models, such as those comprised of subunits or DNA. The objective of this review is to cover the various types of viral vaccines that have been developed and are available for bony fishes, as well as the advantages and challenges that DNA vaccines present for massive administration in a growing aquaculture, possible risks for the environment, the controversy regarding genetically modified organisms and possible acceptance by consumers.

Veterinaria Mexico OA ISSN: 2448-6760 Cite this as: Steffani Hernandez G, Chavez Maya F, Rojas Anaya E, Loza Rubio E, Garcia Espinosa G. Genomic analysis of an atypical Mexican low-pathogenic H5N2 avian influenza virus. Veterinaria Mexico OA. 2016;3(2). doi: 10.21753/vmoa.3.2.363 We analysed the genome of a low-pathogenic avian H5N2 influenza virus isolated from the faeces of experimentally infected Pekin ducks and Leghorn-type chickens to determine its origin and molecular characteristics. The complete genomic sequence was determined using a Sanger-based genome sequencing method and was subsequently characterized by phylogenetic analysis and genetic comparison. The results of this study showed that 8 genomic segments corresponded to an avian influenza virus that were related with strains isolated in Mexico. Investigation of the haemagglutinin gene revealed the presence of few basic amino acids at the cleavage site and lack of a potential N-glycosylation site at position 11. The gene encoding the PB1 protein lacked PB1-F2 and the basic polymerase gene codes for PA-X. In addition, the basic polymerase gene contained the consensus ribosomal frameshifting motif TCC TTT CGT C, which is required for the expression of the PA-X. Molecular characteristics showed that the virus has features of a low-pathogenic H5 influenza virus with the exception of a potential N-glycosylation site at position 11. The genome information for this particular virus will provide a molecular map for further in vivo studies to identify why some influenza viruses can persist in chickens for long periods of time. Such information will be useful in countries such as Mexico, where the virus has been a poultry health problem since 1994 and has the potential to evolve high pathogenicity. Figure 1. Phylogenetic tree of the HA gene of A/chicken/Mexico/2007 (H5N2) Influenza A virus [ ]. The phylogenetic tree was constructed using the neighbour-joining method. The percentages of replicate trees in which the associated taxa clustered together in a bootstrap test (1,000 replicates) are shown next to the branches. The evolutionary distances were computed using the Kimura 2-parameter method. The analysis involved 101 chicken nucleotide sequences. Evolutionary analyses were conducted in MEGA 5.05. A larger triangle size represents a larger number of nucleotide sequences with genetic relationship.

The hepatitis E virus (HEV) is the causative agent of Hepatitis E, an enterically transmitted disease. HEV infections in pigs and humans have been reported worldwide, but data from Mexico are scarce. In the present study, the prevalence of anti-HEV IgG antibodies was investigated in a quite large number of swine from Mexico by means of an ELISA based on a recombinant open reading frame 2 protein of HEV genotype 3. Serum samples from 683 healthy pigs (1–48 months old), collected during 2010–2013 in 109 herds from 48 municipalities located in 9 states in the centre of the country were assayed. A 30.75 % (210/683) of the sera tested were positive, and they were distributed along all the states included in the study. The prevalence of anti-HEV antibodies varied widely between municipalities and herds, and it was higher in pigs 4–6 months of age. No relationships were detected between seroprevalences and farm characteristics. Forty individual faecal samples were analysed by RT-PCR and all resulted negative. These data indicate that HEV infection is widespread in Mexican pigs; thus, representing a potential zoonotic risk for humans. © 2016, Springer Science+Business Media New York.

"Este estudio analizó el genoma de un virus de influenza aviar H5N2, aislado de heces de pollos tipo Leghorn y patos Pekín, infectados experimentalmen - te para conocer su origen y características moleculares. La secuencia comple - ta del genoma del virus se obtuvo por medio del método de secuenciación de Sanger, una vez obtenida la secuencia, se caracterizó mediante compa - ración genética y análisis filogenético. Los resultados del análisis mostraron que los ocho segmentos del genoma están relacionados con virus aislados en México. El análisis del gen de la hemoaglutinina reveló que codifica para pocos aminoácidos básicos en el sitio de corte y quizá carece de un sitio de glicosilación en la posición once. El gen que codifica para la proteína PB1 carece del fragmento PB1-F2, así como en el gen de la PA se encuentra el fragmento PA-X. También se observó, que el gen de la polimerasa contiene la secuencia consenso del ribosoma TCC TTT CGT C, requerida para la ex - presión de PA-X. Las características moleculares demostraron que el virus corresponde a un virus de baja patogenicidad del subtipo H5 con excepción del posible sitio once de glicosilación. La información del genoma para esta cepa del virus, proveerá un mapa molecular para futuros estudios in vivo que puedan contribuir a conocer por qué algunos virus de influenza aviar persisten en los pollos por periodos prolongados. Esta información puede ser de utilidad en países como México, donde el virus ha estado en la avicultura desde 1994 y con el potencial de evolucionar a virus de alta patogenicidad."

Plants have been identified as promising expression systems for the commercial production of vaccines because of the possibility of introducing exogenous genes into them, which permits the development of a new generation of biological products called edible vaccines. The advantages of oral vaccines of this new type are that they induce mucosal, humoral, cellular and protective immunity, they are cheaper, easier to store, distribute and administer, they do not require cold chain management, and some species can be stored for long periods of time without any spoilage and may be administered as purified proteins. Owing to these benefits, plant-produced vaccines represent a valuable option for animal health. The aim of this paper is to present a review of plant-produced vaccines against viruses affecting domestic animals. Some aspects of the feasibility of their use and the immune response elicited by such vaccines are also discussed, as the balance between tolerance and immunogenicity is a major concern for the use of plant-based vaccines.

Antigens derived from various pathogens can readily be synthesized at high levels in plants in their authentic forms. Such antigens administered orally can induce an immune response and, in some cases, result in protection against a subsequent challenge. We here report the expression of rabies virus G protein into carrots. The G gene was subcloned into the pUCpSSrabG vector and then used to transform carrot embryogenic cells by particle bombardment. The carrot cells were selected in liquid medium, a method previously unreported. The presence of the transgene was verified by PCR, and by RT-PCR. By western blot, G protein transgene was identified in 93.3% of adult carrot roots. The G protein was quantified by densitometric analysis (range 0.4-1.2%). The expressed protein was antigenic in mice. This confirms that the carrot is an adequate system for antigen expression.

We characterized the nucleic acid-sensing Toll-like receptors (TLR) of a New World bat species, the common vampire bat (Desmodus rotundus), and through a comparative molecular evolutionary approach searched for general adaptation patterns among the nucleic acid-sensing TLRs of eight different bats species belonging to three families (Pteropodidae, Vespertilionidae and Phyllostomidae). We found that the bat TLRs are evolving slowly and mostly under purifying selection and that the divergence pattern of such receptors is overall congruent with the species tree, consistent with the evolution of many other mammalian nuclear genes. However, the chiropteran TLRs exhibited unique mutations fixed in ligand-binding sites, some of which involved nonconservative amino acid changes and/or targets of positive selection. Such changes could potentially modify protein function and ligand-binding properties, as some changes were predicted to alter nucleic acid binding motifs in TLR 9. Moreover, evidence for episodic diversifying selection acting specifically upon the bat lineage and sublineages was detected. Thus, the long-term adaptation of chiropterans to a wide variety of environments and ecological niches with different pathogen profiles is likely to have shaped the evolution of the bat TLRs in an order-specific manner. The observed evolutionary patterns provide evidence for potential functional differences between bat and other mammalian TLRs in terms of resistance to specific pathogens or recognition of nucleic acids in general.

Using integrative epidemiologic techniques, we studied the changing relationships (beta and phylogenetic beta diversity) of multihost systems and virus associations in bat communities in fragmented landscapes from Chiapas, Campeche and Greater Mexico City. We combined computing applications, molecular detection, and nucleotide sequencing of coronaviruses, hantaviruses, paramyxoviruses and pegiviruses with ecological and phylogenetic analyses. A total of 22 viruses were discovered in 1,067 samples from 42 bat species, representing an estimated 78% of all viral richness in the system. Based on 17 virus genotypes discovered with an equal sampling effort, a total viral richness of 23 genotypes was estimated using a Chao2 statistic model. Using a residual model, we categorized host species and habitat types that are prone to harboring higher viral richness. Positive relationships were found between phylogenetic host diversity and both viral diversity (r = 0.41, p < 0.05) and viral richness (r = 0.51, p < 0.05). The beta diversity (the rate of change) of viral communities was explained by host beta diversity (r = 0.86, p < 0.05). To understand the change in viral and host communities, we partitioned beta diversity in nestedness (species loss) and turnover (compositional dissimilarity) components. In Chiapas, the host beta diversity was explained by the nestedness of species composition, while the phylogenetic host diversity was explained by turnover of the host lineages. Campeche showed a high phylogenetic host nestedness and low host turnover. Beta-diversity and beta-phylogenetic diversity indicated that patterns of local species assemblages and regional abiotic features in human-dominated landscapes are significant drivers of viral community composition. Our study represents the first effort in Mexico to study the relationship between viral diversity in bat communities in modified landscapes to understand host-virus relationships.

To determine the geospatial distribution of bovine paralytic rabies cases transmitted by Desmodus rotundus in the Mexican states of Guanajuato, Querétaro, and San Luis Potosí.This was a cross-sectional epidemiological study based on cases reported during statewide campaigns for the control of bovine paralytic rabies in Guanajuato (2008-2013), Querétaro (2005-2013) and San Luis Potosí (2001-2013). All cases were confirmed by direct immunofluorescence. Maps showing the distribution of cases by year and species were constructed using ArcMap version 10.1. To identify areas where conditions favor the appearance of cases, bioclimatic variables were combined with georeferenced cases using MaxEnt version 3.3.3.Of the 1037 cases recorded, 911 (87.9%) occurred in San Luis Potosí, 82 (7.9%) in Querétaro, and 44 (4.2%) in Guanajuato. Of the total number of cases, 87.4% occurred at altitudes of less than 1500 meters above sea level. In Guanajuato and Querétaro, 77.3% and 42.3% of the cases, respectively, occurred at altitudes greater than 1 500 meters above sea level. Peak incidence was recorded from December to March. The V11 antigenic variant of the virus was the most common (173 cases); it was found in all three states. In the endemic channel, the average number of cases remains within the security zone from January to March but exceeds the median value from April to June. The spatial distribution of cases shows that the disease has spread recently, which correlates with the presence of the vampire bat.Bovine paralytic rabies has spread to areas that were formerly free of the disease. Environmental characteristics and the altitude above sea level do not limit the appearance of cases. Constant monitoring should be conducted for early case detection. Vaccination should take place before the rainy season starts, without waiting for outbreaks to occur.

"Mediante análisis epidemiológicos integrales, se estudiaron la diversidad beta (entre sitios) y la beta filogenética de sistemas multi-hospederos, así como los virus asociados con comunidades de murciélagos en paisajes fragmentados de Chiapas, Campeche y la zona metropolitana de la Ciudad de México. Se combinaron aplicaciones computacionales, técnicas de detección molecular y la secuenciación de nucleótidos de coronavirus, hantavirus, paramyxovirus y pegivirus con análisis ecológicos y filogenéticos. Se descubrieron un total de 22 virus en 1 067 muestras pertenecientes a 42 especies de murciélagos, lo que representa una estimación de 78% de la riqueza viral total. Al aplicar el modelo estadístico Chao2 con datos de los 17 genotipos de virus encontrados con el mismo esfuerzo de muestreo, se calculó una riqueza viral de 23 genotipos. Usando un modelo con residuales, se categorizó a las especies hospederas y los tipos de hábitat que tienen mayor riesgo de asociarse con una mayor riqueza viral. Se encontró una relación positiva entre la diversidad filogenética de los hospederos y la diversidad viral (r = 0.41, P < 0.05) y con la riqueza viral (r = 0.51, P < 0.05). La diversidad beta (tasa de recambio) de las comunidades de virus se explicó por la diversidad beta de los hospederos (r = 0.86, P < 0.05). Para entender los patrones de cambio en las comunidades virales y de hospederos, la diversidad beta se dividió en: un componente de anidamiento —pérdida de especies— y uno de recambio —disimilitud en la composición—. En Chiapas, la diversidad beta de los hospederos se justificó por la anidación de especies, mientras que la diversidad beta filogenética se definió por el recambio de linaje de los hospederos. Campeche mostró altos valores de anidamiento filogenético y bajo recambio de hospederos. La diversidad beta y la diversidad beta filogenética indicaron que los patrones locales de la estructura de las comunidades de hospederos y las características abióticas regionales en paisajes dominados por actividades antropogénicas son un factor importante en la determinación de la composición de comunidades virales. Este estudio representa el primer esfuerzo en México para entender las relaciones hospedero-virus, a través del estudio de las relaciones entre la diversidad viral y las comunidades de murciélagos en paisajes transformados."

Transgenic plants have been employed successfully as a low-cost system for the production of therapeutically valuable proteins, including antibodies, antigens and hormones. Here, we report the expression of the fusion (F) gene of the Newcastle disease virus (NDV) in transgenic maize plants. The expression of the transgene, driven by the maize ubiquitin promoter, caused accumulation of the F protein in maize kernels. The presence of the transgene was verified by Southern and western blots. Feeding chickens with kernels containing the F protein induced the production of antibodies, which conferred protection against a viral challenge. This protection was comparable to that conferred by a commercial vaccine. Possible uses of this plant-based F protein as a potential mucosal vaccine are discussed.

While large-scale dog vaccination campaigns have significantly reduced urban rabies throughout Mexico, reports of sylvatic rabies, including cases of spill-over of bat strains into livestock and humans, are increasing. To improve knowledge of these epidemiological trends, 64 Mexican rabies virus isolates from various host species, have been characterized. Phylogenetic analysis at the viral P locus identified distinct viral strains associated with terrestrial reservoirs (dog, skunk and fox/bobcat) and a variant associated with the insectivorous bat, T. brasiliensis, consistent with prior reports. Of the two distinct clades of viruses associated with the vampire bat reservoir, one comprised just four specimens and formed an outlying group to all other vampire bat rabies isolates including those from South America and the Caribbean, a finding consistent with the early emergence of the vampire bat reservoir in Mexico. Antigenic variation of the vampire bat specimens did not correlate with the main genetic groupings; moreover complete N gene sequence analysis of selected specimens indicated limited variation within the encoded nucleoprotein that could form the basis of antigenic variation. A single isolate recovered from a cat represents a new viral variant not previously identified in North America that probably circulates in a species of insectivorous bat.

Infectious pancreatic necrosis virus (IPNV) is one of the most important viruses in the Pacific salmon Oncorhynchus spp., Atlantic Salmon Salmo salar, and Rainbow Trout O. mykiss industry. This virus has been shown to produce high mortality among salmonid fry and juveniles, and survivors might become carriers. Since 2000, IPNV has affected Mexican Rainbow Trout culture, resulting in considerable economic losses. In the current study, molecular characterization of the VP2 gene of a number of Mexican IPNV isolates was done and the virus's phylogenetic relationships to IPNV reference strains were investigated. The phylogenetic analysis indicated that Mexican IPNV isolates are closely related to strains from the United States and Canada and that all Mexican IPNV isolates belong to genogroup 1. Furthermore, low genetic diversity was found between the Mexican isolates (identity, 95.8-99.8% nucleotides and 95.8-99.6% amino acids). The result of the analysis of the amino acid residues found at positions 217, 221, and 247 (alanine, threonine, and glutamic acid, respectively) could be associated with virulence, although the expression of virulence factors is more complex and may be influenced by the agent and host factors. The high percentage of identity among the VP2 genes from geographically distant IPNV isolates and the evidence of wide distribution in the country might have been facilitated by carrier trout. This hypothesis is supported by the identification of the amino acid threonine at position 221 in all Mexican isolates, a factor related to the carrier state for IPNV, as reported by other studies.

Rabies has been one of the most feared diseases throughout history. Human rabies remains an important public health problem in many developing countries. The WHO reports that more than 55,000 people die of this disease every year. Most of these cases occur in developing countries. In most Latin American countries, the major reservoirs of rabies are the dog and the hematophagous bat (Desmodus rotundus), which is present in the tropical and subtropical areas from Northern Mexico to Northern Argentina and Chile and transmits the disease to cattle. One of the better options for controlling rabies is vaccination. The expression of rabies virus G protein in different plant systems for developing an oral rabies vaccine could reduce costs of production and distribution and would be convenient for developing countries where the disease is endemic.

Background The design of efficient rabies control programs within a geographic area requires an appropriate knowledge of the local epidemiological cycles. In Latin America, there is a geographical overlap of the two main epidemiological cycles: (a) the terrestrial cycle, where the dog is the main terrestrial vector and the principal cause of human transmission; and (b) the aerial cycle, in which the vampire bat Desmodus rotundus is representative in Mexico. This bat is the major sylvatic rabies vector transmitting rabies to cattle. The purpose of this study was to distinguish between the epidemiological cycles of rabies virus (aerial and terrestrial) circulating in Mexico, using restriction fragment length polymorphism (RFLP). Methods Thirty positive rabies isolates were obtained from different species (including humans, domestic, and wildlife animals) and geographical regions. The methodology included the extraction of RNA, and synthesis of cDNA, PCR, and RFLP using four restriction endonucleases. To determine the aerial cycle, BsaW I and BsrG I were utilized, and for terrestrial cycle, BamH I and Stu I. Most of the samples belonged to the aerial and terrestrial cycles, except for two skunk isolates from Northwestern Mexico, which were not cut by any of the enzymes. Results Three different migration patterns were detected: (a) the first was observed in six amplicons, which were cut by BsaW I and BsrG I (aerial cycle); (b) 19 amplified samples were digested with BamH I and Stu I enzymes (terrestrial cycle); and (c) two skunk isolates from Northwest Mexico, were not cut by any of the enzymes utilized in the experiments (hypervariable cycle). Conclusions This concludes that RFLP can be used for the classification of rabies field samples in epidemiological studies. Moreover, it has demonstrated its usefulness, not only for differentiating between the main epidemiological rabies cycles present in Mexico, but also to detect new cycles in wildlife species.

The introduction of exogenous genes into plants permits the development of a new generation of biological products, i.e., edible vaccines. Cereals, especially maize, have been the systems of choice for the expression of antigenic proteins because the proteins can be expressed at high levels in the kernel and stored for prolonged periods without excessive deterioration. The utilization of plant-derived antigens for oral delivery provides an alternative strategy for the control of pathogens in animals compared to the current vaccine administration methods, such as injection. However, there is some doubt about the efficacy of these types of vaccines in polygastric animals due to the features of their digestive system. Here, we report the efficacy of an edible vaccine against rabies evaluated in sheep. Kernels containing different doses of G protein (0.5, 1, 1.5 and 2mg) were given in a single dose by the oral route. Cumulative survival was better in groups that received 2mg of G protein and for the positive control (inactivated rabies vaccine); this observation was supported by the presence of neutralizing antibodies. Animals in the control group died after challenge. The degree of protection achieved for 2mg of G protein was comparable to that conferred by a commercial vaccine. In conclusion, this is the first study in which an orally administered edible vaccine showed efficacy in a polygastric model.

Specific-pathogen-free chickens immunized at 14 days of age with either an inactivated recombinant Newcastle disease virus-LaSota/avian influenza H5 (K-rNDV-LS/AI-H5) vaccine or a killed Newcastle disease/avian influenza whole-virus vaccine (K-ND/AI) were protected from disease when challenged with either A/chicken/Queretaro/14588-19/95 (H5N2), a high pathogenicity avian influenza virus (HPAIV) strain isolated in Mexico in 1995, or with a Mexican velogenic viscerotropic Newcastle disease virus (VVNDV) strain 21 days postvaccination. All nonvaccinated chickens challenged with HPAIV or VVNDV succumbed to disease, while those vaccinated with K-rNDV-LS/AI-H5 or K-ND/AI were protected from severe clinical signs and death. Both vaccines induced hemagglutination-inhibition (HI) antibody responses against NDV and AIV. Antibodies against AIV nucleoprotein were not detected by enzyme-linked immunosorbent assay (ELISA) in birds vaccinated with the inactivated rNDV-LS/AI-H5 vaccine. These chickens became positive for AIV antibodies by ELISA only after challenge with HPAIV. The data clearly indicate that the inactivated rNDV-LS/AI-H5 vaccine confers protection comparable to that of the conventional killed whole-virus vaccine against both NDV and AIV, while still allowing differentiation of infected from vaccinated animals by HI and ELISA tests.

West Nile virus (WNV), a Flavivirus distributed most widely, is presenting lately variable epidemiological and ecological patterns, including an increasing virulence that has already caused over 1000 human deaths in USA. Currently, diagnosis of WNV is achieved mainly by enzyme-linked immunoassays (ELISAs) based on the use of inactivated whole WNV (iWNV) as antigen, although results have to be confirmed by plaque reduction neutralization tests (PRNTs). Expression of WNV envelope recombinant E (rE) protein and its usefulness as ELISA antigen are described. Production of rE was achieved upon infection of Trichoplusia ni insect larvae with a recombinant baculovirus. Once optimized, the rE-based ELISA was validated with a battery of mouse and equine sera characterized previously. Concordance with the iWNV-based ELISA used routinely was good (95%), as it was with the reference PRNT (90%), with specificity of 94.4% and sensitivity of 88.1%. Production of rE protein in insect larvae allows for an easy, low cost and quite large-scale yield of partially purified antigen which is suitable for serological diagnosis of WNV, without the need for manipulation of large quantities of infective virus. © 2010 Elsevier B.V.

SUMMARYWest Nile virus (WNV) was probably introduced in southern and northern Mexico from the USA in two independent events. Since then, WNV activity has been reported in several Mexican states bordering the USA and the Gulf of Mexico, but disease manifestations seen there in humans and equids are quite different to those observed in the USA. We have analysed WNV seroprevalence in asymptomatic, unvaccinated equids from two Mexican states where no data had been previously recorded. WNV IgG antibodies were detected in 31·6% (91/288) of equine sera from Chiapas and Puebla states (53·3% and 8·0%, respectively). Analysis by plaque reduction neutralization test (PRNT) showed good specificity (99·4%) and sensitivity (84·9%) with the ELISA results. Further analyses to detect antibodies against three different flaviviruses (WNV, St Louis encephalitis virus, Ilheus virus) by haemagglutination inhibition (HI) tests on a subset of 138 samples showed that 53% of the 83 HI-positive samples showed specific reaction to WNV. These data suggest continuous expansion of WNV through Mexico.