Dynamics of neutralizing antibodies against Bovine respiratory syncytial virus in a dairy herd from Santa Fe Province, Argentina

Abstract Bovine respiratory syncytial virus (BRSV) is one of the most relevant agents responsi-ble for respiratory disease in cattle from both dairy and beef farms. BRSV is spread by horizontalcontact causing a constant presence of seropositive animals that favors viral circulation throughout the year. Moreover, reinfections with BRSV are frequent between animals regardless of theirage as BRSV does not confer long-lasting protective immunity. Several studies have demonstrated the circulation of BRSV in cattle from different regions of the world; however, little isknown about the dynamics of BRSV infection in cows before and after they begin lactation. Theaim of this work was to study the dynamics of BRSV neutralizing antibodies from birth up to36 months of age in a closed dairy herd of Argentina specifically around the lactation period. Passive maternal antibodies against BRSV started to decrease monthly and became almost undetectable at 8 months of age. We detected two potential infection points at months 11 and 27after birth, in which 30% and 45% of the animals showed seroconversion, respectively. Specifically, an increase in the proportion of seropositive cows after the start of lactation suggests thatthey became reinfected around the time they began lactating. We demonstrate the importanceof understanding BRSV dynamics in a closed dairy herd to review the vaccination schedule ofthe animals to achieve protection against BRSV infection.

Resumen El virus respiratorio sincitial bovino (Bovine respiratory syncytial virus, [BRSV]) es uno de los principales agentes responsables de la enfermedad respiratoria en bovinos, tanto de tambos como de cría. El virus se transmite horizontalmente y causa la presencia constante de animales seropositivos, lo cual favorece la circulación viral a lo largo del ano. A su vez, las reinfecciones por BRSV son frecuentes entre animales independientemente de su edad, dado que el virus no confiere inmunidad protectora a largo plazo. Numerosos estudios han demostrado la circulación de BRSV en bovinos de diferentes regiones del mundo, sin embargo, poco se conoce acerca de la dinámica de infección en vacas antes y después del inicio de la fase de lactancia. El objetivo de este trabajo fue estudiar la dinámica de anticuerpos neutralizantes anti- BRSV en vacas lecheras desde el nacimiento hasta los 36 meses de vida en un tambo cerrado de Argentina, específicamente, en el período de lactancia. Los anticuerpos pasivos específicos para BRSV comenzaron a declinar mensualmente hasta ser casi indetectables a los 6 meses. Detectamos dos potenciales puntos de infección a los meses 11 y 27 luego del nacimiento, momentos en los que el 30 y el 45% de los animales mostraron seroconversión, respectivamente. El incremento en la proporción de vacas seropositivas luego del comienzo de la lactancia sugiere que estas se reinfectaron en el inicio de dicha etapa. Demostramos la importancia de entender la dinámica de circulación del BRSV en un tambo cerrado, a fin de revisar el esquema de vacunación de los animales para que estén protegidos frente a la posible infección por este virus.

Pathogenesis of Bovine alphaherpesvirus 2 in calves following different routes of inoculation / Patogênese do alfaherpesvírus bovino 2 em bezerros inoculados por diferentes vias

Bovine alphaherpesvirus 2 (BoHV-2) is the agent of herpetic mammilitis (BHM), a cutaneous and self-limiting disease affecting the udder and teats of cows. The pathogenesis of BoHV-2 is pourly understood, hampering the development of therapeutic drugs, vaccines and other control measures. This study investigated the pathogenesis of BoHV-2 in calves after inoculation through different routes. Three- to four-months seronegative calves were inoculated with BoHV-2 (107TCID50.mL-1) intramuscular (IM, n=4), intravenous (IV, n=4) or transdermal (TD) after mild scarification (n=4) and submitted to virological, clinical and serological monitoring. Calves inoculated by the IV route presented as light increase in body temperature between days 6 to 9 post-inoculation (pi). Virus inoculation by the TD route resulted in mild inflammatory lesions at the sites of inoculation, characterized by hyperemia, small vesicles, mild exudation and scab formation, between days 2 and 8pi. Virus or viral DNA was detected by PCR in the crusts/swabs collected from lesions of 3 out of 4 animals inoculated TD from day 2 to 8pi. Viremia was detected in 3/4 animals of the IM group (from day 4 to 8pi); in 2/4 animals of the IV group (days 6 and 8pi) but not in the TD group. Calves from all inoculated groups seroconverted to BoHV-2 in titers from 4 to 64, as indicated by virus-neutralizing (VN) assays performed in sera collected at day 15pi. Administration of dexamethasone (Dex) to the inoculated calves at day 48pi, did not result in virus reactivation as indicated by lack of virus detection in the blood and/or in inoculation sites and no increase in VN antibody titers. These results demonstrated that BoHV-2 was able to replicate efficiently in calves following different routes of exposure, produced viremia after IM and IV inoculation and was not reactivated by Dex treatment.(AU)

O alfaherpesvírus bovino 2 (BoHV-2) é um agente etiológico da mamilite herpética (BHM), uma doença cutânea e autolimitante do úbere e tetos de vacas. Pouco se sabe sobre a patogênese do BoHV-2, dificultando o desenvolvimento de medicamentos terapêuticos e vacinas. Este estudo investigou a patogênese do BoHV-2 em bezerros após a inoculação por diferentes vias. Bezerros soronegativos de três a quatro meses foram inoculados com BoHV-2 (107TCID50.mL-1) por via intramuscular (IM, n=4), por via intravenosa (IV, n=4) ou transdérmica (TD, n=4) após escarificação leve e submetidos a monitoramento virológico, clínico e sorológico. Os bezerros inoculados pela via IV apresentaram aumento leve da temperatura corporal entre os dias 6 a 9 pós-inoculação (pi). A inoculação do vírus pela via TD resultou em lesões inflamatórias leves nos locais de inoculação, caracterizadas por hiperemia, pequenas vesículas, exsudação leve e formação de crostas, entre os dias 2 e 8pi. O vírus ou DNA viral foi detectado por PCR nas crostas/swabs coletados de lesões de 3 de 4 animais inoculados TD do dia 2 ao 8pi. Viremia foi detectada em 3/4 dos animais do grupo IM (do dia 4 ao 8pi); em 2/4 animais do grupo IV (dias 6 e 8pi), mas não no grupo TD. Bezerros de todos os grupos inoculados soroconverteram o BoHV-2 em títulos de 4 a 64, conforme indicado por ensaios de vírus-neutralização (VN) realizados em soro coletado no dia 15pi. Administração de dexametasona (Dex) nos bezerros inoculados no dia 48pi, não resultou em reativação do vírus, como indicado pela falta de detecção de vírus no sangue e/ou nos locais de inoculação e pela ausência de aumento nos títulos de anticorpos. Estes resultados demonstraram que o BoHV-2 foi capaz de replicar eficientemente em bezerros seguindo diferentes vias de inoculação, produziu viremia após a inoculação IM e IV e não foi reativado pelo tratamento com Dex.(AU)

Coxiella burnetii associated with BVDV (Bovine Viral Diarrhea Virus), BoHV (Bovine Herpesvirus), Leptospira spp, Neospora caninum, Toxoplasma gondii and Trypanosoma vivax in reproductive disorders in cattle / Coxiella burnetii associada ao BVDV (Vírus da Diarreia Viral Bovina), BoHV (Vírus do Herpes Bovino), Leptospira spp, Neospora caninum, Toxoplasma gondii e Trypanosoma vivax em distúrbios reprodutivos em bovinos

Abstract This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens associated with reproductive disorders in cattle in four Brazilian states, including the zoonotic agent Coxiella burnetii. The used tests were Virus Neutralization Assay for IBR and BVD, Microscopic Agglutination Test for Leptospira spp., Indirect Fluorescent Antibody Test (IFAT) for C. burnetii and Toxoplasma gondii, and Enzyme-Linked Immunosorbent Assay for Neospora caninum and Trypanosoma vivax. Seropositivity for C. burnetii was 13.7% with titers from 128 to 131,072; 57.8% for BoHV-1, with titers between 2 and 1,024; 47.1% for BVDV-1a, with titers from 10 to 5,120; 89.2% for N. caninum; 50% for T. vivax; and 52.0% for Leptospira spp., with titers between 100 to 800 (the following serovars were found: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona and Icterohaemorrhagiae); 19.6% for T. gondii with titer of 40. This is the first study that has identified C. burnetii in cattle associated with BoHV and BVDV, N. caninum, Leptospira spp., T. gondii and T. vivax. Thus, future studies should be conducted to investigate how widespread this pathogen is in Brazilian cattle herds.

Resumo Este é um estudo transversal para avaliar a presença de anticorpos em ruminantes contra patógenos selecionados e associados a distúrbios reprodutivos em bovinos de quatro estados brasileiros, incluindo o agente zoonótico Coxiella burnetii. Os testes utilizados foram Teste de Vírus-Neutralização para BoHV e BVDV, teste de Aglutinação Microscópica para Leptospira spp., Reação de Imunofluorescência Indireta for C. burnetii e Toxoplasma gondii, e Ensaio de Imunoabsorção Enzimática para Neospora caninum e Trypanosoma vivax. A soropositividade para C. burnetii foi de 13,7% com títulos de 128 a 131.072; 57,8% para BoHV-1, com títulos entre 2 a 1.024; 47,1% para BVDV-1a, com títulos de 10 a 5.120; 89,2% para N. caninum; 50% para T. vivax; e 52,0% para Leptospira spp., com títulos entre 100 a 800 (sorovares encontrados: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona e Icterohaemorrhagiae) 19,6% para T. gondii com título de 40. Este é o primeiro estudo que evidencia a participação de C. burnetii em bovinos associada ao Vírus da Rinotraqueíte bovina infecciosa e da diarreia viral bovina, N. caninum, Leptospira spp., T. gondii e T. vivax em bovinos. Desta forma, futuros estudos devem ser conduzidos a fim de investigar o quão disseminado se encontra este patógeno em rebanhos bovinos brasileiros.

Bovine respiratory syncytial virus seroprevalence and risk factors in feedlot cattle from Córdoba and Santa Fe, Argentina / Seroprevalencia del virus respiratorio sincicial bovino y factores de riesgo asociados en engordes a corral de Córdoba y Santa Fe, Argentina

Bovine respiratory syncytial virus (BRSV) is one of the causative agents of respiratory disease in cattle all over the world, leading to important economic losses. The aim of this work was to determine the seroprevalence of BRSV in feedlot cattle of Argentina and the risk factors associated with the disease. Results showed a high individual seroprevalence of 78.64% (95% confidence interval adjusted -#91;CI-#93; =66.55-90.75%) against the virus. Positive association was found between the presence of high BRSV neutralizing antibody titers, and the following risk factors: cattle age, source of animals, presence of clinical respiratory signs and herd size. This work contributes to updating the understanding of its epidemiology in Argentinean feedlots and poses the need for reevaluating vaccination strategies against this virus in order to control infection and its impact on productivity.

El virus respiratorio sincicial bovino (BRSV) es uno de los agentes causantes de enfermedad respiratoria bovina a nivel mundial, conduciendo a importantes pérdidas económicas. El objetivo de este trabajo fue determinar la seroprevalencia del BRSV en bovinos de engorde a corral de Argentina y estudiar los factores de riesgo asociados. Los resultados mostraron una elevada seroprevalencia individual del 78,64% (IC = 66,55-90,75%) contra el virus. Se encontró una asociación positiva entre la presencia de altos títulos de anticuerpos neutralizantes contra BRSV y los factores de riesgo: edad del ganado, origen de los animales, presencia de signos clínicos respiratorios y el tamaño del rebano. Este trabajo contribuye en gran medida a la comprensión de la epidemiología en los establecimientos de engorde a corral de Argentina y plantea la necesidad de revaluar las estrategias de vacunación contra este virus con el fin de controlar la infección y su impacto en la producción.

Passive immunity to control Bovine coronavirus diarrhea in a dairy herd in Argentina / Inmunidad pasiva en el control de la diarrea por coronavirus bovino en un rodeo lechero de Argentina

Bovine coronavirus (BCoV) is a viral enteric pathogen associated with calf diarrhea worldwide being, in Argentina, mostly detected in dairy husbandry systems. The aim of the present work was to study if maternal IgG1 antibodies (Abs) to BCoV acquired by colostrum intake modulate the development of BCoV infection in calves reared in a dairy farm in Argentina. Thirty Holstein calves were monitored during their first 60 days of age. Animals were classified into two groups depending on their initial BCoV IgG1 Ab titers. The "failure of passive transfer" (FPT) group had significantly lower IgG1 Abs to BCoV than the "acceptable passive transfer" (APT) group of calves (log10 1.98 vs. 3.38 respectively) (p<0.0001). These differences were also observed when the total protein levels in both groups were compared (p = 0.0081). Moreover, 71% (5/7) of calves from the FPT group showed IgG1 seroconversion to BCoV compared to 29.4% (5/17) of animals from the APT group. Regarding viral circulation, BCoV was detected in 10% (3/30) of all calves and BCoV IgG1 Ab seroconversion was detected in 42% of the total animals showing that almost half of the calves were infected with BCoV. In conclusion, calves with high titers of specific BCoV IgG1 (≥1024) were mostly protected against viral infection, while animals with low titers of IgG1 (<1024) were mostly infected with BCoV. IgG1 Abs from colostrum origin are critical for prevention of BCoV infection.

El coronavirus bovino (Bovine coronavirus, BCoV) es un enteropatógeno viral asociado a la diarrea neonatal del ternero. El objetivo del presente trabajo fue estudiar si los anticuerpos IgG1 anti-BCoV adquiridos pasivamente mediante el calostro modulan la infección por BCoV en terneros de un rodeo lechero de Argentina. Se monitorearon 30 terneros raza Holstein durante los primeros 60 días de vida. Estos animales fueron clasificados en dos grupos según sus niveles de IgG1 anti-BCoV maternales: grupo con transferencia de inmunidad pasiva aceptable (APT) y grupo con fallas en la transferencia pasiva (FPT). Este último grupo tenía un título de IgG1 significativamente menor comparado con el primer grupo (log10 1,98 vs. 3,38, respectivamente; p< 0,0001). La misma diferencia se observó cuando se compararon los niveles de proteínas séricas totales (p = 0,0081). Además, el 71% (5/7) de los terneros del grupo FPT mostró seroconversión de IgG1, mientras que el 29,4% (5/17) de los terneros del grupo APT la mostró. Con respecto a la circulación viral, se detectó BCoV en el 10% (3/30) de los terneros así como también seroconversión de IgG1 en el 42% del total de los animales, lo que evidencia que aproximadamente la mitad de los terneros se infectaron con BCoV. Este estudio mostró que los terneros con altos títulos de IgG1 específica (≥ 1.024) estuvieron mayormente protegidos contra la infección con BCoV, mientras que los animales con títulos bajos de IgG1 (< 1.024) estuvieron predispuestos a la infección. Esto confirma que los anticuerpos IgG1 calostrales son críticos para la prevención de la infección por este agente viral.

Bovine Herpesvirus 4 in Parana State, Brazil: case report, viral isolation, and molecular identification

Bovine Herpesvirus 4 (BoHV-4) is a member of Gammaherpesvirinae sub-family and belongs to genus Rhadinovirus. This virus has been associated with different clinical manifestations and research activity has put forward a strong correlation among virus infection, postpartum metritis, and abortion. The goal of this work was to characterize a virus strain isolate from a cow’s uterine outflow. From swabs drawn of uterine secretion, a virus strain was isolated and characterized by its cytopathology, morphology, and molecular biology approaches. In culture there was CPE development, characterized mainly by long strands with several small balloons along them, radiated from infected cells. Electron microscopy analysis revealed virus particles that had icosahedrical capsid symmetry surrounded by a loose envelope, typical of a herpesvirus. A 2,571 bp PCR product after HindIII digestion generated four fragments, whose base pair composition were 403, 420, 535, and 1,125 bp. Restriction enzymes HindIII and BamHI generated the expected diagnostic bands as well as a 2,350 bp hypermolar fragment as a result of BamHI treatment to demonstrate that agent was a bovine herpesvirus 4, appertaining to DN-599 group.

Teat papillomatosis associated with bovine papillomavirus types 6, 7, 9, and 10 in dairy cattle from Brazil

This study describes the clinical, histopathological, and virological characterization of teat papillomatosis from Brazilian dairy cattle herds. Four types of bovine papillomavirus were identified (BPV6, 7, 9, and 10); one of these (BPV7) is being detected for the first time in Brazilian cattle.

Ultrastructural study of bovine papillomavirus during outbreaks in Brazil / Estudio ultraestructural del virus papiloma bovino durante brotes en Brasil

Papillomaviruses, belonging to the Papillomaviridae family, are small oncogenic viruses, causing papillomas and fibropapillomas in the mucosal and cutaneous epithelia of several animals. In bovine species, thirteen types (BPV 1-13) were characterized to date. In this study, the occurrence of papillomatosis in four outbreaks in cattle herds, coming from Brazilian states were registered. The papillomatous lesions were found located in the teats, udders, head and neck. Under the transmission electron microscope, by the negative staining technique, it was possible to visualized rounded-format papillomavirus, with icosahedral symmetry, characterized as "full" and "empty" particles, measuring on average 60 nm in diameter, in all the 40 samples observed of skin lesion fragments. The ultrathin sections revealed the presence of groups of viral, intranuclear, rounded particles measuring 35 nm in diameter and tubular particles with a diameter of 35-39 nm. At immunoelectron microscopy technique, positivity obtained was marked by the presence of aggregates of viral particles formed by the antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction showed colloidal gold particles evenly distributed over the surface of the virus. These results showed the importance of the transmission electron microscopy techniques in the diagnosis of bovine papillomatosis that can be used in routine procedures to identify viral agent of this important disease.

Los virus del papiloma que pertenecen a la familia Papillomaviridae son pequeños virus oncogénicos que causan papilomas y fibropapilomas en epitelio cutáneo y mucoso de distintas especies de animales. En el ganado vacuno, trece tipos (BPV- 1-13) se caracterizaron hasta el momento. En este estudio, se documenta la ocurrencia de cuatro brotes de papilomatosis en los rebaños de ganado, procedentes de estados brasileños. Las lesiones papilomatosas se localizaron en los pezones, la ubre, la cabeza y el cuello. Al microscopio electrónico de transmisión, en la técnica de tinción negativa fueran visualizadas partículas del virus del papiloma redondeadas, con simetría icosaédrica, caracterizadas como "llenas" y "vacías", midiendo unos 60 nm de diámetro en todas las 40 muestras de fragmentos de lesión de piel estudiado. Los cortes ultra finos mostraron la presencia de grupos de partículas virales, intranucleares redondeadas con 35 nm de diámetro y tubulares 35-39 nm de diámetro. En la técnica de microscopía inmunoelectrónica, la positividad obtenida se caracterizó por la presencia de agregados de partículas virales formadas por la interacción antígeno-anticuerpo. En la aplicación de la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo mostró partículas de oro coloidal distribuidos de manera uniforme sobre la superficie del virus. Estos resultados muestran la importancia de las técnicas de microscopía electrónica de transmisión en el diagnóstico de papilomatosis bovina, que pueden ser utilizados en los procedimientos de rutina para la identificación del agente viral causante de esta importante enfermedad.

Rabies in humans and non-human in the state of Pará, Brazilian Amazon

We evaluate the relationship of positive cases of rabies with the continuing expansion of livestock production, and analyse the trends of this zoonosis in human population in the state of Pará, Brazilian Amazon. The distribution of rabies cases was recorded between 1999 and 2004. Of 148 cases of rabies, 21% were in humans and 79% in non-human mammals. The rapid growth in livestock numbers seems to be associated with the increase of positive cases in bovine livestock transmitted by vampire bats. This idea is supported by positive and significant relationship of both events in time (p < 0.01), but failed when spatial distribution among regions of the state was considered. However, rabies cases tend to occur toward the northeastern of the state of Pará, where rabies cases are proportionally five times greater than other mesoregions, suggesting that increased livestock production may influence the increase of this zoonosis.

[Molecular and histopathological findings on induced pulmonary lesions by bovine viral diarrhea virus]

The bovine viral diarrhea virus [BVDV] is one of the bovine respiratory system pathogens that represses lungs immunity and raises the pathogenicity of other bacteria and viruses. The aim of the present study was to evaluate the effect of BVDV in lung tissue. In this study, 30 bovine lung tissues were used. Histopathologic analysis showed different lesions in lung tissues. RT-PCR test results and histopathologic observations were studied for each sample and compared. Kai Square Test was used to study any possible association between the existence of BVD virus and incidence of four common lesions: 1- Interestitial pneumonia, 2- bronchitis and bronchiolitis, 3- edema, emphysema and atelectasis and 4- bronchopneumonia. If the assumptions of X2 test were not applicable, the Fisher Exact Test was used. The RT-PCR test for BVDV results was positive for 10 lung samples. 5 BVD positive cases were involved with interestitial pneumonia, while 10 BVD negative cases were not affected. Finally, in the studied lung tissues, only the 3 + degree or severe interstitial pneumonia was significantly associated with the existence of BVD virus. No significant relation was seen regarding other possible lesions, which may have resulted from the probable roles of bacteria or other viruses and toxic factors that were not studied in this project

Clinical signs, diagnosis, and case reports of Vaccinia virus infections

Vaccinia virus is responsible for a zoonosis that usually affects cattle and human beings in Brazil. The initial clinical signs of the infection are focal red skin areas, fever, and general symptoms similar to those of a cold. Then, pustules and ulcerated lesions surrounded by edema and erythema follow, as well as local lymphadenopathy that can last for weeks. Cure and healing of the lesions occur over several weeks, leaving a typical scar in the skin of people and animals affected. The infection definitive diagnosis is made through morphological characterization of the virus by use of electron microscopy, followed by PCR for specific viral genes. Since 1963, circulating orthopoxviruses in infectious outbreaks in several regions of Brazil have been reported. Later, the etiological agent of those infections was characterized as samples of Vaccinia virus. In addition, the widespread use of those viruses in research laboratories and mass vaccination of militaries have contributed to increase the cases of those infections worldwide. Thus, several epidemiological and clinical studies are required, as well as studies of viral immunology, public health, and economic impact, because little is known about those Vaccinia virus outbreaks in Brazil.

Genetic characterization of rabies virus isolated from bovines and equines between 2007 and 2008, in the States of São Paulo and Minas Gerais / Caracterização genética de vírus da raiva isolado de bovinos e equinos entre 2007 e 2008, nos Estados de São Paulo e Minas Gerais

INTRODUCTION: Rabies is an acute disease of the central nervous system and is responsible for the deaths of thousands of humans, wild animals and livestock, particularly cattle, as well as causing major economic losses. This study describes the genetic characterization of rabies virus variants that circulate in Desmodus rotundus populations and are transmitted to herbivores. METHODS: Fifty rabies virus isolates from bovines and equines in the States of São Paulo and Minas Gerais, Brazil, were genetically characterized and compared with sequences retrieved from GenBank. RESULTS: Two clusters (I and II) with mean nucleotide identities of 99.1 and 97.6 percent were found. The first of these contained nearly all the samples analyzed. Lineages from other Brazilian states grouped in cluster II. CONCLUSIONS: Analysis of the amino acid sequences of the N proteins revealed the existence of genetic markers that may indicate possible variations between geographic regions, although the biologically active regions are conserved within the species over space and time.

INTRODUÇÃO: A raiva é uma doença aguda do sistema nervoso central e é responsável por mortes de milhares de humanos, animais silvestres e animais de criação - especialmente bovinos - além de causar elevadas perdas econômicas. Este trabalho descreve a caracterização genética das variantes do vírus da raiva que circulam em populações de Desmodus rotundus e são transmitidas aos herbívoros. MÉTODOS: Cinquenta isolados de vírus da raiva de bovinos e equinos provenientes dos Estados de São Paulo e Minas Gerais, Brasil, foram caracterizadas geneticamente e comparadas com sequências recuperadas do GenBank. RESULTADOS: Dois clusters, I e II, apresentando identidades médias de nucleotídeos de 99,1 e 97,6 por cento, foram obtidos, sendo o primeiro composto de quase a totalidade das amostras analisadas. Linhagens de outros estados do Brasil "clustered" no II. CONCLUSÕES: A análise das sequências de aminoácidos da proteína N revelou que existem marcadores genéticos que podem determinar uma possível regionalidade embora as regiões biologicamente ativas apresentem-se conservadas dentro das espécies ao longo do tempo e espaço.

Polimorfismos de BoLA-DRB3 en el bovino sintético colombiano Lucerna y asociación con conteo de células somáticas y mastitis / BoLA-DRB3 Polymorphism in Colombian synthetic cattle Lucerna and association with somatic cell counts and mastitis

En bovinos se han determinado asociaciones de BoLA-DRB3 con enfermedades infecciosas y con el onteo de células somáticas. En este estudio fue analizada la raza sintética colombiana Lucerna, parael microsatélite DRB3 intrón 2. Fueron identificados 24 alelos por amplificación con PCR y geles de poliacrilamida, encontrando frecuencias alélicas desde 0.02 hasta 0.26. El alelo mas frecuente en la población analizada fue el 161, y el menos frecuente 159. Ambas subpoblaciones afectadas y no afectadas por mastitis, presentaron frecuencias similares para la mayoría de los alelos. La población no se encuentra en equilibrio H.W (Hardy- Weinberg). (p <0.05), y presentó déficit de heterocigóticos. El valor medio de recuento de células somáticas transformado (log2 (CCS / 100000 ) + 3), para la población total fue de 4.8 (+/- 2.89) y para cada subpoblación de afectadas y no afectadas fue de 5.76 y 3.86, respectivamente. Se estudiaron las asociaciones potenciales entre los alelos BoLA-DRB3, el recuento de células somáticas y la mastitis clínica, y se encontrando una asociación significativa (p<0.05) entre el alelo 191 con un alto conteo de células somáticas. No se encontró asociación significativa entre el alelo 191 y la enfermedad, pero si hubo significancia estadística (p<0.05) para la asociación de la mastitis con las variables hato y número de partos.

Bovine papillomavirus type 2 detection in the urinary bladder of cattle with chronic enzootic haematuria

The bovine papillomavirus type 2 (BPV-2) involvement in the aetiology of chronic enzootic haematuria associated to bracken fern ingestion has been suggested for a long time. However, a few reports have shown the presence of the BPV-2 in urinary bladder tumors of cattle. The aim of this study was to investigate the presence of the BPV-2 infection in the urinary bladder of cattle with chronic enzootic haematuria in Brazilian cattle herds. Sixty-two urinary bladders were collected from adult cattle in beef herds from the north region of the state of Paraná, Brazil. According to clinical and pathological finds the specimens were distributed in three groups: the group A was constituted by 22 urinary bladders with macroscopic lesions collected at necropsy of cattle with clinical signs of chronic enzootic haematuria; the group B by 30 urinary bladders with macroscopic lesions collected in a slaughterhouse of cows coming from bracken fern-endemic geographical region; and the group C (control) by 10 urinary bladders without macroscopic lesions collected from asymptomatic cattle in a bracken fern-free geographical region. By a semi-nested polymerase chain reaction (PCR) assay, with an internal control, a fragment of the BPV-2 L1 gene with 386 bp length was amplified in 36 (58 percent) urinary bladder. The rate of BPV-2 positive urinary bladders was 50 percent (11/22) for group A, 80 percent (24/30) for group B, and 10 percent (1/10) for group C (control). The rate of the positive results found in groups A and B that included urinary bladder samples with macroscopic lesions was 67 percent (35/52) and the detection of the BPV-2 in both groups was significantly higher (P < 0.05) than in the control group. RFLP with Rsa I and Hae III enzymes evaluated the specificity of the BPV-2 amplicons. The PCR internal control that amplified a 626 bp fragment of the ND5 gene of the bovine mitochondrial genome was amplified in all analyzed samples and excluded false-negatives or invalid results in the semi-nested PCR...

Febre catarral maligna em bovinos do Mato Grosso do Sul e de São Paulo / Malignant catarrhal fever in cattle from the states of Mato Grosso do Sul and São Paulo, Brazil

São descritos a epidemiologia, os sinais clínicos, os achados de necropsia e a histopatologia de seis casos de febre catarral maligna (FCM) em bovinos de 5 fazendas localizadas nos estados de Mato Grosso do Sul e São Paulo. A doença ocorreu em bovinos de ambos os sexos e com idades variando de 4 meses a 11 anos. Os índices de morbidade variaram de 0,25 por cento a 6.6 por cento e a letalidade foi de 100 por cento. A evolução clínica foi aguda (2-3 dias) em seis casos e crônica em um (3 meses). O diagnóstico presuntivo de FCM nos casos descritos neste relato foi baseado nos sinais clínicos, achados de necropsia e confirmados pela histopatologia. Os principais sinais clínicos nos casos agudos foram febre, corrimento mucopurulento pelas fossas nasais, opacidade da córnea, sialorréia, úlceras em várias superfícies mucosas e distúrbios nervosos. O bovino do caso crônico mostrou opacidade da córnea e distúrbios neurológicos. Os principais achados de necropsia incluíam hiperemia e lesões diftéricas em várias superfícies epiteliais e a histopatologia consistiu de vasculite, focos de infiltrado mononuclear multifocal em vários órgãos e necrose de superfícies epiteliais.

Lengua Azul: Aislamiento y caracterización del virus e identificación de vectores en el noreste argentino / Bluetongue: isolation and characterization of the virus and identification of vectors in northeastern Argentina

To establish if BTV was circulating in Argentina, 94 bovines from the Santo TomÚ and Ituzaingó Departments of Corrientes Province were sampled every 30-60 days during 14 months. Red blood cells from those animals that showed seroconvertion with a c-ELISA were processed for virus isolation by inoculation in embryonated chicken eggs and cell cultures. Cells with CPE were positive by direct and indirect immunofluorescence with BTV specific reagents. These samples examined by electron microscopy showed virus particles with BTV morphological characteristics. Blood samples and tissue culture supernantants were positive by RT-PCR technique with primers corresponding to the segment 3 of the BTV genome. Haematophagous insects were captured in one farm using light traps and Culicoides insignis Lutz was the predominant species detected. This is the first isolation of BTV in Argentina from northeastern bovines without any disease symptom.

Rotavirus bovino grupo A: diagnóstico y caracterización antigénica de cepas circulantes en la República Argentina, 1994-1999 / Group A bovine rotavirus: diagnosis and antigenic characterization of strains circulating in the Argentine Republic, 1994-1999

Group A Bovine Rotavirus (BRV) has been identified as a major cause of neonatal diarrhea in cattle. The study was aimed to determine the prevalence of BRV and to antigenically characterize the G-types of circulating strains in dairy and beef herds in Argentina. A total of 1129 stool samples from diarrheic calves was analyzed from 1994 to 1999. The samples were initially screened for RV by ELISA and PAGE, and then G-typed using monoclonal antibodies (Mab) directed against G1, G2, G3, G6 and G10-specific epitopes. Forty percent (452/1129) of the samples were positive for RV by ELISA, while 24.7 (279/1129) were also positive for PAGE. VP7 was detected in the 70.5 (319/452) of the positive samples using a broadly reactive Mab (C60); 32.6 (104/319) were G6, 15.4 (49/319) were G10, and 6 (19/319) were G1. However, 46.1 (147/319) of the samples remained untypable. Rotavirus diarrhea prevalences were comparable in beef and dairy herds (87.3 and a 74.4, respectively). Finally, G6 was the most prevalent G-type circulating in beef herds while G10 prevailed in dairy herds. A better understanding of RV epidemiology will contribute to the optimization of current vaccines and prevention programs of RV diarrhea in calves.

Caracterización molecular de cepas del virus de la fiebre aftosa aisladas durante los años 1993-1994 en la Argentina / Molecular characterization of aphthous fever virus isolated during the years 1993-1994 in Argentina

Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.

Detection of poxvirus in cattle associated with human cases in the State of Rio de Janeiro: preliminary report

This preliminary report describes human and cow cases of poxvirus that recently ocurred in the State of Rio de Janeiro. The electron microscopic findings were consistent with parapoxviral and orthopoxviral infection. Orthopoxvirus strains were isolated from human and cow cases. Detailed viral characterization by means of genetical techniques is under investigation. Based on these informations, poxviral diseases should be also considered an emerging viral zoonosis that can affect human beings.