Quantitation of Rabies Virus in Various Bovine Brain Structures.

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.

Swine virome on rural backyard farms in Mexico: communities with different abundances of animal viruses and phages.

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.

Lack of association between amino acid sequences of the bovine leukemia virus envelope and varying stages of infection in dairy cattle.

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.

Virulence of infectious pancreatic necrosis virus (IPNV) isolates from Mexico.

Infectious pancreatic necrosis virus (IPNV) causes economic losses in Mexican rainbow trout industry. In this study, virulence and genetic fingerprints of Mexican IPNV isolates was investigated for the first time. Two Mexican IPNV isolates were analyzed in rainbow trout fry and the Sp strain was included as high virulence. One of the Mexican IPNV isolate was obtained from diseased fish and the other from fish without clinical signs. The infection was performed using a standardized immersion. Clinical signs were observed at 4 days post infection in fry group infected with strain Sp, two days earlier than in trout infected with IPNV isolates Mexican. Severe lesions were found in 100% of the individuals of Sp group, but only in 25% of each isolated Mexican group. Results suggest that Mexican IPNV isolates are pathogenic, but less virulent than strain Sp. The amino acid motif residues of both Mexican isolates, corresponded to a subclinical disease. Nevertheless, the accumulated motility observed in the field, suggest that other factors play a role in the virulence of the disease.

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.

Leukocyte Profiles Reflect Geographic Range Limits in a Widespread Neotropical Bat.

Quantifying how the environment shapes host immune defense is important for understanding which wild populations may be more susceptible or resistant to pathogens. Spatial variation in parasite risk, food and predator abundance, and abiotic conditions can each affect immunity, and these factors can also manifest at both local and biogeographic scales. Yet identifying predictors and the spatial scale of their effects is limited by the rarity of studies that measure immunity across many populations of broadly distributed species. We analyzed leukocyte profiles from 39 wild populations of the common vampire bat (Desmodus rotundus) across its wide geographic range throughout the Neotropics. White blood cell differentials varied spatially, with proportions of neutrophils and lymphocytes varying up to six-fold across sites. Leukocyte profiles were spatially autocorrelated at small and very large distances, suggesting that local environment and large-scale biogeographic factors influence cellular immunity. Generalized additive models showed that bat populations closer to the northern and southern limits of the species range had more neutrophils, monocytes, and basophils, but fewer lymphocytes and eosinophils, than bats sampled at the core of their distribution. Habitats with access to more livestock also showed similar patterns in leukocyte profiles, but large-scale patterns were partly confounded by time between capture and sampling across sites. Our findings suggest that populations at the edge of their range experience physiologically limiting conditions that predict higher chronic stress and greater investment in cellular innate immunity. High food abundance in livestock-dense habitats may exacerbate such conditions by increasing bat density or diet homogenization, although future spatially and temporally coordinated field studies with common protocols are needed to limit sampling artifacts. Systematically assessing immune function and response over space will elucidate how environmental conditions influence traits relevant to epidemiology and help predict disease risks with anthropogenic disturbance, land conversion, and climate change.

Fecal virome composition of migratory wild duck species.

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.

Hologenomic adaptations underlying the evolution of sanguivory in the common vampire bat.

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.

Viral Communities Among Sympatric Vampire Bats and Cattle.

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.

Bats, Primates, and the Evolutionary Origins and Diversification of Mammalian Gammaherpesviruses.

Gammaherpesviruses (γHVs) are generally considered host specific and to have codiverged with their hosts over millions of years. This tenet is challenged here by broad-scale phylogenetic analysis of two viral genes using the largest sample of mammalian γHVs to date, integrating for the first time bat γHV sequences available from public repositories and newly generated viral sequences from two vampire bat species (Desmodus rotundus and Diphylla ecaudata). Bat and primate viruses frequently represented deep branches within the supported phylogenies and clustered among viruses from distantly related mammalian taxa. Following evolutionary scenario testing, we determined the number of host-switching and cospeciation events. Cross-species transmissions have occurred much more frequently than previously estimated, and most of the transmissions were attributable to bats and primates. We conclude that the evolution of the Gammaherpesvirinae subfamily has been driven by both cross-species transmissions and subsequent cospeciation within specific viral lineages and that the bat and primate orders may have potentially acted as superspreaders to other mammalian taxa throughout evolutionary history. IMPORTANCE: It has long been believed that herpesviruses have coevolved with their hosts and are species specific. Nevertheless, a global evolutionary analysis of bat viruses in the context of other mammalian viruses, which could put this widely accepted view to the test, had not been undertaken until now. We present two main findings that may challenge the current view of γHV evolution: multiple host-switching events were observed at a higher rate than previously appreciated, and bats and primates harbor a large diversity of γHVs which may have led to increased cross-species transmissions from these taxa to other mammals.

Detection and phylogenetic analysis of bovine papillomavirus in cutaneous warts in cattle in Tamaulipas, 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.

Les papillomes sont rencontrés plus fréquemment chez les bovins que chez n'importe quelle autre espèce domestiques. L'agent causal de la papillomatose bovine est un virus appartenant à la famille Papillomaviridae. Dans l'état mexicain de Tamaulipas le virus est considéré comme un problème sérieux et a empêché l'exportation de bovin vers les États-Unis d'Amérique, causant ainsi des pertes économiques importantes. Étant donné le manque d'information concernant les sous-types de papillomavirus qui infectent les bovins au Mexique, l'objectif de l'étude était de déterminer les sous-types présents dans l'état de Tamaulipas. Cinquante-deux verrues ont été analysées par réaction d'amplification en chaine par la polymérase (ACP) à l'aide d'amorces amplifiant le gène E7 du papillomavirus bovin (PVB). Les produits de l'ACP ont été séquencés afin de différencier les sous-types PVB-1 et PVB-2. La qualité du séquençage fut déterminée à l'aide du logiciel MEGA 6.0. La comparaison des séquences obtenues pour l'état de Tamaulipas avec celles des types connus de PVB par l'algorithme MUSCLE a permis de démontrer que 53 % étaient des PVB-1 et 47 % de PVB-2. La distribution des deux sous-types chez les bovins était homogène. La présente étude démontre la présence de PVB-1 et PVB-2 chez les bovins de Tamaulipas et constitue le premier rapport sur la caractérisation moléculaire des papillomes au Mexique.(Traduit par Docteur Serge Messier).

Prevalence of Hepatitis E Virus (HEV) Antibodies in Mexican Pigs.

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.

Comportamiento epidemiológico de la rabia paralítica bovina en la región central de México, 2001-2013 / Epidemiological trends in bovine paralytic rabies in central Mexico, 2001-2013

OBJETIVO:Determinar la distribución geoespacial de los casos de rabia paralítica bovina transmitida por Desmodus rotundus en los estados de Guanajuato, Querétaro y San Luis Potosí, México. MÉTODOS: Estudio epidemiológico transversal a partir de los casos notificados por las campañas estatales de control de la rabia paralítica bovina en Guanajuato (2008-2013), Querétaro (2005-2013) y San Luis Potosí (2001-2013). Se confirmaron los casos por inmunofluorescencia directa. Se elaboraron mapas de distribución de los casos por año y por especie, mediante el software ArcMap versión 10.1. Para identificar áreas con condiciones apropiadas para la presencia de casos se combinaron las variables bioclimáticas con los casos georreferenciados, mediante el programa MaxEnt versión 3.3.3. RESULTADOS: Se registraron 1 037 casos, de los cuales, 911 (87,9%) ocurrieron en San Luis Potosí, 82 (7,9%) en Querétaro y 44 (4,2%) en Guanajuato. Del total, 87,4% ocurrió en alturas menores de 1 500 msnm. En Guanajuato y Querétaro, 77,3% y 42,3% de los casos, respectivamente, ocurrieron en alturas mayores de 1 500 msnm. Los meses de mayor incidencia fueron de diciembre a marzo. La variante antigénica viral V11 fue la más frecuente (173 casos), presente en los tres estados estudiados. En el canal endémico, el promedio de casos se mantiene en la zona de seguridad de enero a marzo, pero de abril a junio excede la mediana. La distribución espacial de los casos muestra la diseminación reciente de la enfermedad, lo que coincide con la presencia del murciélago vampiro. CONCLUSIONES: La rabia paralítica bovina se ha extendido a regiones anteriormente libres de esta enfermedad. Las características ambientales y la altura sobre el nivel del mar no limitan la presentación de casos. Se debe mantener un monitoreo constante para la detección oportuna de casos. La vacunación se debe realizar antes del comienzo de las lluvias, sin esperar la aparición de brotes.

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

The evolution of bat nucleic acid-sensing Toll-like receptors.

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.

A novel endogenous betaretrovirus in the common vampire bat (Desmodus rotundus) suggests multiple independent infection and cross-species transmission events.

The Desmodus rotundus endogenous betaretrovirus (DrERV) is fixed in the vampire bat D. rotundus population and in other phyllostomid bats but is not present in all species from this family. DrERV is not phylogenetically related to Old World bat betaretroviruses but to betaretroviruses from rodents and New World primates, suggesting recent cross-species transmission. A recent integration age estimation of the provirus in some taxa indicates that an exogenous counterpart might have been in recent circulation.

[Epidemiological trends in bovine paralytic rabies in central Mexico, 2001-2013]. / Comportamiento epidemiológico de la rabia paralítica bovina en la región central de México, 2001-2013.

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Molecular characterization of the VP2 gene of infectious pancreatic necrosis virus (IPNV) isolates from Mexico.

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.

Prolonged excretion of a low-pathogenicity H5N2 avian influenza virus strain in the Pekin duck.

H5N2 strains of low-pathogenicity avian influenza virus (LPAIV) have been circulating for at least 17 years in some Mexican chicken farms. We measured the rate and duration of viral excretion from Pekin ducks that were experimentally inoculated with an H5N2 LPAIV that causes death in embryonated chicken eggs (A/chicken/Mexico/2007). Leghorn chickens were used as susceptible host controls. The degree of viral excretion was evaluated with real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) using samples from oropharyngeal and cloacal swabs. We observed prolonged excretion from both species of birds lasting for at least 21 days. Prolonged excretion of LPAIV A/chicken/ Mexico/2007 is atypical.

Viral vaccines for bony fish: past, present and future.

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.