Bovine Rabies Research Articles

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.

Landscape and cattle management attributes associated with the incidence of Desmodus rotundus attacks on cattle

Bovine paralytic rabies, transmitted by Desmodus rotundus, causes economic losses for Latin American livestock producers. Although the total number of reported cases has decreased markedly due to vaccination campaigns, economic losses can be significant for smallholders. Identifying risk factors for vampire bat attacks on cattle could help guide preventive control efforts in specific areas. The objective of this study was to analyze the relationship between the incidence of D. rotundus attacks on cattle and various landscape features and cattle management practices in northeastern Puebla, Mexico. We visited 61 properties with cattle between May and October 2011, in which we quantified the incidence of attacks on cattle and described landscape and cattle management attributes. The following features were described within a 200-m radius around the nightly resting site of cattle: topography, percent forest cover, and number of forest fragments, rivers and streams, tree lines, buildings, roads and highways. With these variables, we built models that were assumed to describe the levels of human interference, prey availability, and movement facilitation for bats. The human interference model (number of buildings plus number of roads) best explained the incidence of attacks on cattle. According to our results, the cattle with the highest vulnerability to attacks by vampire bats are those resting at night in less confined sites, far away from human activity. Results of this study suggest that rabies vaccination campaigns on cattle should focus on fragmented areas with few livestock management practices.

Bovine rabies: economic loss and its mitigation through antirabies vaccination

ABSTRACT: Rabies is among the most common neurological disease in cattle in Brazil, causing significant economic losses. Data on the economic impact of rabies in livestock are available in several countries. However, in Brazil, these data focus mainly on the public health point o view, emphasizing the costs related to the prevention of rabies in humans, in dogs, or wildlife. Specific studies carried out in different regions of Brazil indicate critical economic losses caused by rabies in cattle in this country. However, the studies on the losses caused by the disease in cattle lack a detailed analysis of the affected rural properties based on data from official disease control agencies. The objective of this work was to evaluate the economic impact of bovine rabies, and its mitigation through antirabies vaccination in rural properties in Mato Grosso do Sul, Midwestern Brazil.

The emergence of vampire bat rabies in Uruguay within a historical context.

Pathogen spillover from wildlife to humans or domestic animals requires a series of conditions to align with space and time. Comparing these conditions between times and locations where spillover does and does not occur presents opportunities to understand the factors that shape spillover risk. Bovine rabies transmitted by vampire bats was first confirmed in 1911 and has since been detected across the distribution of vampire bats. However, Uruguay is an exception. Uruguay was free of bovine rabies until 2007, despite high-cattle densities, the presence of vampire bats and a strong surveillance system. To explore why Uruguay was free of bovine rabies until recently, we review the historic literature and reconstruct the conditions that would allow rabies invasion into Uruguay. We used available historical records on the abundance of livestock and wildlife, the vampire bat distribution and occurrence of rabies outbreaks, as well as environmental modifications, to propose four alternative hypotheses to explain rabies virus emergence and spillover: bat movement, viral invasion, surveillance failure and environmental changes. While future statistical modelling efforts will be required to disentangle these hypotheses, we here show how a detailed historical analysis can be used to generate testable predictions for the conditions leading to pathogen spillover.

The impact of paralytic bovine rabies transmitted by vampire bats in Latin America and the Caribbean.

The effective control of dog rabies in Latin America is justifiably seen as a major success in the struggle to limit this devastating zoonosis. However, rabies remains a problem, due to the presence of the virus in bat populations throughout the region, including vampire bats. Vampire bats obtain nutrition exclusively through consuming blood by biting mammals and birds. This makes the species a highly efficient vector of the rabies virus, responsible for sporadic outbreaks of rabies in human populations and numerous cases of rabies in livestock. This, in turn, causes economic losses to the farming industry in countries throughout the region. For over four decades, efforts to control rabies have been directed at controlling the reservoir species and vaccinating cattle. However, this approach has not eliminated rabies in livestock. A major barrier to innovation in vampire rabies control is a lack of consistent surveillance to establish the extent of the problem. This precludes any calculation of its cost to the economy or the cost of potential solutions, such as vaccinating livestock. This paper outlines the problem of livestock rabies in Latin America and considers factors that influence the economic cost of potential solutions to this continuing challenge to human and livestock health.

Glycoprotein-G-gene-based molecular and phylogenetic analysis of rabies viruses associated with a large outbreak of bovine rabies in southern Brazil.

A large outbreak of hematophagous-bat-associated bovine rabies has been occurring in Rio Grande do Sul (RS), the southernmost Brazilian state, since 2011, with official estimates exceeding 50,000 cattle deaths. The present article describes a genetic characterization of rabies virus (RABV) recovered from 59 affected cattle and two sheep, from 56 herds in 16 municipalities (2012-2016). Molecular analysis was performed using the nucleotide (nt) and predicted amino acid (aa) sequences of RABV glycoprotein G (G). A high level of nt and aa sequence identity was observed among the examined G sequences, ranging from 98.4 to 100%, and from 97.3 to 100%, respectively. Likewise, high levels of nt and aa sequence identity were observed with bovine (nt, 99.8%; aa, 99.8%) and hematophagous bat (nt, 99.5%; aa, 99.4%) RABV sequences from GenBank, and lower levels were observed with carnivore RABV sequences (nt, 92.8%; aa, 88.1%). Some random mutations were observed in the analyzed sequences, and a few consistent mutations were observed in some sequences belonging to cluster 2, subcluster 2b. The clustering of the sequences was observed in a phylogenetic tree, where two distinct clusters were evident. Cluster 1 comprised RABV sequences covering the entire study period (2012 to 2016), but subclusters corresponding to different years could be identified, indicating virus evolution and/or introduction of new viruses into the population. In some cases, viruses from the same location obtained within a short period grouped into different subclusters, suggesting co-circulation of viruses of different origins. Subcluster segregation was also observed in sequences obtained in the same region during different periods, indicating the involvement of different viruses in the cases at different times. In summary, our results indicate that the outbreaks occurring in RS (2012 to 2016) probably involved RABV of different origins, in addition to a possible evolution of RABV isolates within this period.

Geographic distribution of <em>Desmodus rotundus</em> in Mexico under current and future climate change scenarios: Implications for bovine paralytic rabies infection

Veterinaria México OA ISSN: 2448-6760Cite this as:Zarza H, Martínez-Meyer E, Suzán G, Ceballos G. Geographic distribution of Desmodus rotundus in Mexico under current and future climate change scenarios: Implications for bovine paralytic rabies infection. Veterinaria México OA. 2017;4(3). doi: 10.21753/vmoa.4.3.390.Climate change may modify the spatial distribution of reservoirs hosting emerging and reemerging zoonotic pathogens, and forecasting these changes is essential for developing prevention and adaptation strategies. The most important reservoir of bovine paralytic rabies in tropical countries, is the vampire bat (Desmodus rotundus). In Mexico, the cattle industry loses more than $2.6 million US dollar, annually to this infectious disease. Therefore, we predicted the change in the distribution of D. rotundus due to future climate change scenarios, and examined the likely effect that the change in its distribution will have on paralytic rabies infections in Mexico. We used the correlative maximum entropy based model algorithm to predict the potential distribution of D. rotundus. Consistent with the literature, our results showed that temperature was the variable most highly associated with the current distribution of vampire bats. The highest concentration of bovine rabies was in Central and Southeastern Mexico, regions that also have high cattle population densities. Furthermore, our climatic envelope models predicted that by 2050–2070, D. rotundus will lose 20 % of its current distribution while the northern and central regions of Mexico will become suitable habitats for D. rotundus. Together, our study provides an advanced notice of the likely change in spatial patterns of D. rotundus and bovine paralytic rabies, and presents an important tool for strengthening the National Epidemiological Surveillance System and Monitoring programmes, useful for establishing holistic, long-term strategies to control this disease in Mexico.Figure 4. Modelled suitability for future distribution of Desmodus rotundus according to Global Climate Model GFDL-CM3 for two time periods (2050 and 2070), and two Representative Concentration Pathways (RCP 4.5 and 8.5). Left-hand column shows suitability values, with blue indicating more suitable conditions.

Geographical Analysis for Detecting High-Risk Areas for Bovine/Human Rabies Transmitted by the Common Hematophagous Bat in the Amazon Region, Brazil.

BackgroundThe common hematophagous bat, Desmodus rotundus, is one of the main wild reservoirs of rabies virus in several regions in Latin America. New production practices and changed land use have provided environmental features that have been very favorable for D. rotundus bat populations, making this species the main transmitter of rabies in the cycle that involves humans and herbivores. In the Amazon region, these features include a mosaic of environmental, social, and economic components, which together creates areas with different levels of risk for human and bovine infections, as presented in this work in the eastern Brazilian Amazon.MethodologyWe geo-referenced a total of 175 cases of rabies, of which 88% occurred in bovines and 12% in humans, respectively, and related these cases to a number of different geographical and biological variables. The spatial distribution was analyzed using the Kernel function, while the association with independent variables was assessed using a multi-criterion Analytical Hierarchy Process (AHP) technique.FindingsThe spatiotemporal analysis of the occurrence of rabies in bovines and humans found reduction in the number of cases in the eastern state of Pará, where no more cases were recorded in humans, whereas high infection rates were recorded in bovines in the northeastern part of the state, and low rates in the southeast. The areas of highest risk for bovine rabies are found in the proximity of rivers and highways. In the case of human rabies, the highest concentration of high-risk areas was found where the highway network coincides with high densities of rural and indigenous populations.ConclusionThe high-risk areas for human and bovine rabies are patchily distributed, and related to extensive deforested areas, large herds of cattle, and the presence of highways. These findings provide an important database for the generation of epidemiological models that could support the development of effective prevention measures and controls.

Rabies in Costa Rica: Documentation of the Surveillance Program and the Endemic Situation from 1985 to 2014.

This is the first comprehensive epidemiological analysis of rabies in Costa Rica. We characterized the occurrence of the disease and demonstrated its endemic nature in this country. In Costa Rica, as in other countries in Latin America, hematophagous vampire bats are the primary wildlife vectors transmitting the rabies virus to cattle herds. Between 1985 and 2014, a total of 78 outbreaks of bovine rabies was reported in Costa Rica, with documented cases of 723 dead cattle. Of cattle outbreaks, 82% occurred between 0 and 500 meters above sea level, and seasonality could be demonstrated on the Pacific side of the country, with significantly more outbreaks occurring during the wet season. A total of 1588 animal samples, or an average of 55 samples per year, was received by the veterinary authority (SENASA) for rabies diagnostic testing at this time. Of all samples tested, 9% (143/1588) were positive. Of these, 85.6% (125/1588) were from cattle; four dogs (0.3% [4/1588]) were diagnosed with rabies in this 30-year period. Simultaneously, an extremely low number (n = 3) of autochthonous rabies cases were reported among human patients, all of which were fatal. However, given the virus' zoonotic characteristics and predominantly fatal outcome among both cattle and humans, it is extremely important for healthcare practitioners and veterinarians to be aware of the importance of adequate wound hygiene and postexpositional rabies prophylaxis when dealing with both wild and domestic animal bites.

Spatial and Temporal Description of Laboratory Diagnosis of Bovine Rabies in the State of Rio Grande do Sul, Brazil

Background: Rabies remains one of the most important zoonosis worldwide and in Brazil. In the Brazilian state of Rio Grande do Sul (RS) the disease is one of the main causes of death in bovine herds. The confirmation of suspected rabies cases upon clinical examination depends on specific laboratory diagnosis. The Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF) is the reference laboratory in RS to diagnose rabies. The objective of the present study was to present a spatial-temporal description of diagnostic procedures of rabies in cattle in the years 2011 to 2015 in RS.Materials, Methods & Results: The results of diagnosis procedures of rabies in cattle carried out in the Laboratory of Virology, IPVDF, between January 2011 and December 2015 were listed. Direct immunofluorescence (DIF) and biological assays for confirming rabies cases were used. Cattle population density per area of a municipality, as well as date, species involved, location of suspected case, and diagnosis were obtained from exams and from official rabies record, and input to a spreadsheet file. Spatial distribution maps of cattle rabies confirmed in laboratory and cattle population density per municipality were constructed using the software TerraView 4.2.2 (INPE). In the study period, 998 samples of brains of cattle presenting neurological symptoms compatible with rabies, of which 582 (58%) were confirmed in the laboratory. The frequency of positive cases on a yearly basis varied between 41 and 65%, with statistically lower number of positives in 2011 (P > 0.05). The annual incidence of rabies for a population of 10,000,000 bovines was 33, 88, 112, 108, and 79, respectively, for the years 2011 to 2015, in that order. A positive correlation was observed between the number of samples analyzed and the incidence of positive cases in the study period, and statistically significant difference in incidence between years (P > 0.05). Samples were from 175 municipalities, of which 114 (65%) had at least one confirmed rabies case and 61 did not have any. The five municipalities that sent most samples for analysis were Viamão (n = 66), Montenegro (n = 40), Camaquã (n = 39), Gravataí (n = 38), and São Lourenço do Sul (n = 37). The spatial distribution of rabies in cattle in a year did not follow a defined pattern, though most cases confirmed in the laboratory occurred in the greater Porto Alegre region and in southeast RS.Discussion: High incidence of rabies was observed in 2012 and 2013, and in the two following years the number of cases confirmed in laboratory remained high, compared with 2011. The high incidence of rabies cases confirmed in laboratory correlated with the number of samples analyzed. The fall in vaccine coverage during the study period may also have prompted the increase in suspected cases and in the number of samples examined in the laboratory. A large number of the municipalities presenting rabies cases in cattle was in the greater Porto Alegre region and in southeast RS, which are areas of high and intermediate risk for the disease, according to a previous epidemiological modeling study. The spatial distribution of positive cases was not associated with the cattle population density. The large number of rabies in cattle in RS underscores the importance of the disease and the high risk of exposure, both to humans and animals. The results presented herein may help define control and prevention actions against rabies in RS.

Virus isolation in cell culture for confirmatory diagnostic of rabies in bovine specimens

ABSTRACT: This study investigated the suitability of virus isolation (VI) in mouse neuroblastoma cells (N2A) and baby hamster kidney cells (BHK-21) as a confirmatory test for diagnosis of bovine rabies. Fourty-eight brain samples from cattle suspected of rabies were initially submitted to fluorescent antibody test (FAT) and mouse inoculation test (MIT) for routine diagnostic. Subsequently, these specimens were submitted to three protocols of VI in each cell line: a single 24h or 72h passage (T1, T2), or three 48h passages (T3). The FAT and MIT combined detected 32/48 positive samples, from which MIT detected 32 and FAT 31. The average time required for final MIT results was 12.3 days (8 - 21). VI in BHK-21 cells provided definitive, positive results in 100% of the samples in 72h (T2) and in 96.9% after three 48h passages (T3). VI in N2A cells yielded positive results in 100% in 72h (T2) and in 93.7% of samples after three 48h passages (T3). Sensitivity, specificity, positive and negative predictive values were 100% in T2 in N2A and BHK-21 cells, and the Kappa value was excellent in both cells (k=1). A single 24h passage (T1) in both cell lines performed poorly, detecting less than 40% of the positive samples. Taking together, these results indicate that VI in both cell lines, especially in BHK-21 cells that grow faster and are much easier to maintain, does represent an adequate alternative for MIT as a confirmatory test for rabies diagnostic in bovine specimens, yielding reliable results in reduced time.

Bovine rabies in Greece in 2013

Rabies has a worldwide distribution and continues to remain a considerably significant cause of death in many developing and third world countries. Rabies carries a noteworthy economic burden on endemic...

Retrospective Study on Rabies in Laellay Adiabo District of Tigrey, Ethiopia

Retrospective study from records of human health center and veterinary clinic of the district was conducted to determined prevalence of rabies in the last five years (2009-2013) in Laellay Adiabo district of Tigrey, Ethiopia. A questionnaire was distributed to 60 randomly selected individuals. Of the animal case recorded, within the past five years, a total of 154 suspected cases of rabies were recorded at the veterinary clinic. 41.56% (64/154) of bovine rabies cases were presented to the veterinary clinic. On the other hand, of the human case recorded at the health center, a total of 62 suspected cases of rabies were recorded on the case book of the health center. In the last five years, out of the sixty two suspected human rabies cases, 31.66% (19/62) was presented to the health center of the district in the year 2013. Similarly males were more affected than females and 43.55% of the suspected cases were registered in the age range of greater than 15 years. Questionnaire survey revealed that 25% (15/60) had observed rabies cases in human and in different animals in their surroundings. 100% respondents replied that the transmission of rabies was through dog bite. 58.33%, 8.33% and 33.34% of the study individuals’ diagnosis rabid animals by salivation, barking the dogs and eating inanimate objects, respectively.

Histopathology and immunohistochemistry of tissues outside central nervous system in bovine rabies.

We performed a histopathological and immunohistochemical study of tissues outside the central nervous system in 48 cases of bovine rabies confirmed by direct immunofluorescence and/or immunohistochemistry (IHC) of the central nervous system. In the bovines of this study, mononuclear inflammation in all ganglia (trigeminal, spinal, stellate, and celiac) and adrenal medulla was observed. This injury also occurred in 85 % of neuro-pituitaries in 55 % of pars intermediate and 15 % of the pars distalis of pituitary evaluated. IHC was positive in 92.31 % of lumbar spinal ganglia, 90.9 % of trigeminal ganglia, stellate ganglia of 41.67 and 16.67 % of the celiac ganglia. One of the evaluated adrenal (1/17) showed strong immunohistochemical labeling in the cytoplasm of pheochromocytes. The pituitary IHC was positive in one case in the neurohypophysis (1/20) and in one case in the pars intermedia of the adenohypophysis (1/20). Data from this study indicate that in suspected cases of rabies, besides the complex pituitary rete mirabile and trigeminal ganglion, the evaluation of other ganglia, particularly the lumbar spinal, and adrenal may also contribute to the diagnosis and understanding of the clinical presentation and pathogenesis of the disease in bovines.

Epidemiologia molecular de surto de raiva bovina na região central do Rio Grande do Sul, 2012

A raiva é uma doença infecciosa do sistema nervoso central de mamíferos causada pelo vírus da raiva (RabV), geralmente, transmitido pela mordedura de animais infectados. No Brasil, os morcegos hematófagos Desmodus rotundus são as principais fontes de infecção do RabV para bovinos e equinos. Este artigo descreve uma investigação epidemiológica e molecular de surtos de raiva ocorridos na região central do Rio Grande do Sul, entre maio e agosto de 2012. Nesse período, 45 casos suspeitos de raiva foram relatados em 22 pequenos rebanhos, localizados dentro de um raio de 4,7km, no município de Pinhal Grande. Desses, 32 amostras foram submetidas para diagnóstico da raiva, sendo que o RabV e/ou antígenos virais foram identificados em 27 amostras. Em um segundo momento, 11 amostras foram submetidas à transcrição reversa/reação em cadeia da polimerase (RT-PCR) para o gene da nucleoproteína (N) do RabV, seguido de sequenciamento nucleotídico e análise filogenética. Sete das 11 amostras apresentaram sequências nucleotídicas idênticas e uma apresentou mutação sinônima, não-codificante, indicando uma provável origem comum dos vírus. Por outro lado, três amostras apresentaram mutações que resultaram em alterações de aminoácidos, sugerindo uma origem diferente do vírus. Esses resultados sugerem que RabV de diferentes origens/linhagens co-circulam na região e foram envolvidos nos surtos descritos. Investigações sobre a circulação de ambos os genótipos em morcegos na região estão em andamento.

Bovine diseases causing neurological signs and death in Mexican feedlots

The number of large feedlot operations, similar to that of USA and Canada, has notably increased in Mexico in the last three decades. Clinical and laboratory diagnoses of neurological diseases in feedlot cattle are crucial in Mexico and Central America because of the high incidence of bovine paralytic rabies (BPR). Because of its zoonotic potential, BPR must be promptly diagnosed and differentiated from other bovine neurological diseases such as thrombotic meningoencephalitis (TME), polioencephalomalacia (PEM) and botulism. More recently, BPR and botulism have been diagnosed with increasing frequency in Mexican feedlots. Neither BPR nor botulism has relevant gross lesions, thus post-mortem diagnosis without laboratory support is impossible. Herein, we describe five outbreaks of neurological diseases in Mexican feedlots in which BPR, botulism and PEM were diagnosed either independently or in combination. A diagram illustrating the most conspicuous pathologic findings and ancillary laboratory test required to confirm the diagnoses of these neurological diseases in feedlot cattle is proposed.

Predictive qualitative risk model of bovine rabies occurrence in Brazil

Bovine rabies remains endemic in Brazil and despite control efforts, the disease still spreads insidiously. The main vector is the hematophagous bat, Desmodus rotundus. The present work aimed to create a predictive qualitative model of the occurrence of bovine rabies in each municipality in 25 of the 27 Brazilian States. The risk of rabies transmission from bats to bovine was estimated using decision-tree models of receptivity and vulnerability. Questionnaires, which covered a number of questions related to the surveillance of possible risk factors, such as bovine rabies outbreaks in the previous year, the presence of bat roosts, bat rabies positivity and environmental changes, were sent to the local veterinary units of each State. The bovine density and geomorphologic features were obtained from national databases and geographic information systems. Of the 433 municipalities presenting bovine rabies outbreaks in 2010, 178 (41.1%) were classified by the model as high risk, 212 (49.0%) were classified as moderate risk, 25 (5.8%) were classified as low risk, whereas the risk was undetermined in 18 municipalities (4.1%). An ROC curve was built to determine if the risk evaluated by the model could adequately discriminate between municipalities with and without rabies occurrence in future years. The risk estimator for the year 2011 was classified as moderately accurate. In the future, these models could allow the targeting of rabies control efforts, with the adoption of control measures directed to the higher risk locations and the optimization of the field veterinary staff deployment throughout the country. Additionally, efforts must be made to encourage continuous surveillance of risk factors.

Bovine rabies in Turkey: patterns of infection and implications for costs and control

The incidence of rabies in livestock is an important factor for estimating the economic impact of the disease, but obtaining reliable data is hindered by inadequate surveillance. In order to understand the contribution of livestock rabies to the overall burden of disease, the rabies incidence in cattle was investigated in detail for Turkey between 2008 and 2011. Data were compiled on cattle numbers, samples submitted for rabies diagnosis, vaccinated animals and positive rabies cases in animals for seven regions in Turkey. Rabies incidence in cattle fluctuated annually and differed between regions from 0·10 to 3·87 cases/100 000 animals. The positive influence of compensation schemes was observed. Livestock losses were conservatively estimated at around $250 000 international dollars per annum, although in areas where compensation schemes are not operating this could be an underestimate of the economic burden. Vaccination of cattle remains an option for disease prevention, although oral rabies vaccination through aerially distributed baits should be implemented to prevent the further spread of fox-mediated rabies, which could result in much greater economic costs.

Antigenic and genetic characterization of rabies virus isolates from Uruguay

After 25 years without any reported cases of rabies in Uruguay, the northern region of the country experienced an epizootic of bovine paralytic rabies in October 2007. The outbreak affected bovines and equines, and the main source of infection was the bat Desmodus rotundus, the only hematophagous species in the country. From October 2007 to July 2008, 42 bovine, 3 equine and 120 chiropteran samples were submitted to the National Veterinary Diagnostic Laboratory for rabies testing. A total of 12 samples (7 bovine, 2 equine and 3 from D. rotundus) were positive by the fluorescent antibody test, and viruses were isolated by the mouse inoculation test. The objective of this study was to compare the antigenic and genetic characteristics of these isolates and three isolates from insectivorous bats from other regions. Antigenic typing using a panel of eight monoclonal antibodies identified all 12 viruses as variant 3 (AgV3), a variant associated with D. rotundus. Two isolates from insectivorous bats (Tadarida brasiliensis and Molossus sp.) were characterized as antigenic variant 4 (AgV4) while the third, from Myotis sp., could not be characterized using this panel as its reactivity pattern did not match that of any of the known antigenic variants. Partial N-gene sequences (nt 149–1420) of these isolates were aligned with homologous sequences derived from GenBank by the CLUSTAL/W method and used to build a neighbor-joining distance tree with the Kimura 2-parameter model. All 12 isolates were genetically grouped into the D. rotundus cluster as they shared 100% identity. In the phylogenetic analysis, the three isolates from insectivorous bats segregated into three clusters: one related to T. brasiliensis, one to Myotis sp. and the other to Lasiurus sp., although the isolate associated with the latter came from a Molossus sp. specimen. These results indicate that AgV3 was associated with the outbreak of bovine paralytic rabies in Uruguay. This is the first report of rabies virus having been detected in non-hematophagous bats in this country.

Domestic microwave processing for rapid immunohistochemical diagnosis of bovine rabies.

The present study describes the use of a microwave processing protocol for the rapid histopathological and immunohistochemical diagnosis of bovine rabies. Immunohistochemistry has been used for rabies diagnosis in formalin-fixed tissue with satisfactory results, although the time to diagnosis is considerably longer than that with direct immunofluorescence. The protocol provided a provisory histopathological rabies diagnosis in approximately three and half hours and the immunohistochemical diagnosis was available after six hours. The protocol achieved 100% correlation with direct immunofluorescence and is a promising method, particularly in situations in which only material in formalin is available for diagnosis or when the refrigeration or transportation of biological material is difficult.