Ultrastructure of sperm 'tail stump' defect in wild boar.

Sperm 'tail stump' defect was found in ejaculates of a wild boar maintained in captivity. It was in good physical condition, the testes and genital tract were found to be of normal size and consistency. There was no evidence of macroscopic abnormalities at the clinical analysis and at necropsy. The volume and concentration of the semen samples obtained by electroejaculation were lower than normal. The slides examined contained a high level of abnormal spermatozoa (52.7%). The most frequent morphological finding was a droplet-like form attached to the base of the head or a very short stump. The non-stumped spermatozoa had no normal tail but a shortened one. Analysing the histological structure with light microscopy, no ring of spermatozoa was observed lining the lumen of the seminiferous tubules and the characteristically cellular structure was not conserved. The ultrastructural examination evidenced a disorganisation of the normal tubular structure of the flagellum, with lost of regular pattern of the axial bundle of fibrils and the mitochondrial helix. The origin of this abnormality is unknown.

Two-step (culture and PCR) diagnostic approach for differentiation of non-T. foetus trichomonads from genitalia of virgin beef bulls in Argentina.

Preputial fluids from 567 virgin Angus and Hereford bulls, 1-2 years old, were inoculated into Sutherland medium, and approximately 8.4% produced cultures with a protozoan suggestive of Tritrichomonas foetus. Under brightfield microscopy, large numbers of single-celled motile organisms with multiple anterior flagellae, a posterior flagellum, axostyle, and a visible undulating membrane were detectable. Motility was jerky and rolling, as described for T. foetus. Air-dried smears of cultures stained with Giemsa or Diff-Quick + iodine revealed an organism similar to T. foetus, although somewhat more rounded. Several organisms appeared to have four anterior flagellae. Scanning electron microscopy (5000x) of representative samples revealed four anterior flagellae on most organisms, and an axostyle that was consistently longer than that seen in T. foetus. Using pan-trichomonal primers and T. foetus-specific primers in a polymerase chain reaction (PCR) assay, amplification products of 372bp were detected in all virgin bull isolates, but only with the pan-trichomonal primers. Positive control isolates of T. foetus yielded amplification products of the expected size (372 and 347bp) with the two sets of primers, respectively. We conclude that these protozoa are not T. foetus, and note the similarity of these findings with those reported earlier in North American beef cattle. Because in several countries there is no legal treatment for bovine trichomonosis, veterinarians recommend slaughter of bulls with positive preputial cultures. The existence of easily mis-identified non-T. foetus trichomonads in the bovine prepuce suggests that the current "gold standard" diagnostic test (culture of preputial scrapings or washings) should be augmented with a more specific confirming test, such as the PCR employed in this study.

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

To establish if BTV was circulating in Argentina, 94 bovines from the Santo Tomé 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.

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.

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 TomU 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.(AU)

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 Tomé 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.

The human breast cancer cell line IIB-BR-G has amplified c-myc and c-fos oncogenes in vitro and is spontaneously metastatic in vivo.

IIB-BR-G is an undifferentiated, highly heterogeneous, hormone receptor negative human breast cancer cell line previously established in our laboratory from a patient's primary tumor. An in vitro growing cell line (IIB-BR-G) and a xenotransplanted tumor growing in nude mice (IIB-BR-G(NUDE)) were derived. To further characterize these systems, immunocytochemical analysis was performed for differentiation antigens (PEM 200 kDa, CEA, NCA 90 kDa), blood-group related antigens (Le(x), sTn), oncogenes and tumor suppressor gene products (Her-2/neu protein, p53), metastasis-related cathepsin D and CD63/5.01 Ag, and the chemokine monocyte chemotactic protein 1 (MCP-1). Expression of markers was heterogeneous in these different systems. Previously reported karyotypic analysis has shown extensive chromosomal alterations including double min. Searching for oncogene amplification, we detected augmented copy number of c-myc and c-fos, the last one with two rearranged fragments. No amplification was found for c-erbB-2 in the cell line or in IIB-BR-G(NUDE), although this oncogene was amplified in the patient's primary tumor DNA. The differences observed between the patient's tumor, the cell line and the IIB-BR-G(NUDE) tumors are probably due to clonal expansion of cell variants not present in the original tumor. Electron microscopy of IIB-BR-G growing cells revealed epithelial characteristics with abundant dense granules, presumably secretory, distributed all over the cytoplasm and great nuclear pleomorphism. In vitro, IIB-BR-G cells showed a significant number of invading cells by Matrigel assay. After nearly 40 sequential subcutaneous passages of the original xenograft through nude mice, 80% of recipients developed spontaneous metastases, primarily to the lung and lymph nodes. Since this experimental model allowed to analyze changes produced in cancer cells from the primary tumor during adaptation to in vitro and in vivo growth, our results provide novel insights on the behaviour of hormone independent metastatic breast cancer.

[Response of guinea pigs to vaccination with parainfluenza virus 3]. / Respuesta en cobayos a la vacunación con virus Parainfluenza-3.

An inactivated vaccine was prepared with Parainfluenza-3 virus strain LQ-514 and strains of Pasteurella hemolytica and P. multocida, suspended in oil adjuvant. The virus had been isolated from 30-60 day old calves during an epidemic of Pneumonia. The vaccine was tested in guinea pigs aged 1 to 2 months. The antibody response and the virus titres in organs after the challenge were the parameters studied. Hemagglutination inhibition antibodies were first detected 14 days after vaccination and reached maximum titres at day 28. The challenge was done at day 34, and a secondary antibody response was observed 72 hours later, which reached its peak the following day. Virus could be isolated from lung samples of control animals at day 3, 4 and 5 after infection. Moreover, viral antigens and particles were observed in the same samples by immunofluorescence and electron microscopy, respectively. In contrast, all three methods failed to demonstrate the presence of virus in organs of immunized guinea pigs after the challenge.

Respuesta en cobayos a la vacunación con virus Parainfluenza-3 / Response of guinea pigs to vaccination with Parainfluenza-3 virus

Se elaboró una vacuna inactivada con la cepa LQ-514 del virus Parainfluenza-3 aislado en brotes de neumonía en terneros de 30 a 60 días de edad y cepas de Pasteurella hemolítica y P. multocida. En este trabajo se estudió la posibilidad de evaluar dicha vacuna con adyuvante oleoso en cobayos de 1 a 2 meses de edad, a través de la respuesta de anticuerpos y aislamiento de virus luego de la descarga. En los animales vacunados pudieron detectarse anticuerpos inhibidores de la hemoaglutinación a partir del día 14 p.i. llegando a títulos máximos el día 28 p.i. A los tres días de la descarga se evidenció una respuesta de tipo secundario en los animales vacunados, alcanzando el máximo al cuarto día. El virus pudo aislarse en cultivos celulares a partir de pulmón de animales no inmunizados los días 3,4 y 5 post descarga, evidenciándose además su presencia por inmunofluorescencia directa y por microscopía electrónica. Por el contrario, en los animales vacunados y descargados no pudo aislarse virus de pulmón ni ponerse en evidencia por inmunofluorescencia directa o microscopía electrónica

Respuesta en cobayos a la vacunación con virus Parainfluenza-3 / Response of guinea pigs to vaccination with Parainfluenza-3 virus

Se elaboró una vacuna inactivada con la cepa LQ-514 del virus Parainfluenza-3 aislado en brotes de neumonía en terneros de 30 a 60 días de edad y cepas de Pasteurella hemolítica y P. multocida. En este trabajo se estudió la posibilidad de evaluar dicha vacuna con adyuvante oleoso en cobayos de 1 a 2 meses de edad, a través de la respuesta de anticuerpos y aislamiento de virus luego de la descarga. En los animales vacunados pudieron detectarse anticuerpos inhibidores de la hemoaglutinación a partir del día 14 p.i. llegando a títulos máximos el día 28 p.i. A los tres días de la descarga se evidenció una respuesta de tipo secundario en los animales vacunados, alcanzando el máximo al cuarto día. El virus pudo aislarse en cultivos celulares a partir de pulmón de animales no inmunizados los días 3,4 y 5 post descarga, evidenciándose además su presencia por inmunofluorescencia directa y por microscopía electrónica. Por el contrario, en los animales vacunados y descargados no pudo aislarse virus de pulmón ni ponerse en evidencia por inmunofluorescencia directa o microscopía electrónica (AU)

Respuesta en cobayos a la vacunación con virus Parainfluenza-3. / [Response of guinea pigs to vaccination with parainfluenza virus 3]

An inactivated vaccine was prepared with Parainfluenza-3 virus strain LQ-514 and strains of Pasteurella hemolytica and P. multocida, suspended in oil adjuvant. The virus had been isolated from 30-60 day old calves during an epidemic of Pneumonia. The vaccine was tested in guinea pigs aged 1 to 2 months. The antibody response and the virus titres in organs after the challenge were the parameters studied. Hemagglutination inhibition antibodies were first detected 14 days after vaccination and reached maximum titres at day 28. The challenge was done at day 34, and a secondary antibody response was observed 72 hours later, which reached its peak the following day. Virus could be isolated from lung samples of control animals at day 3, 4 and 5 after infection. Moreover, viral antigens and particles were observed in the same samples by immunofluorescence and electron microscopy, respectively. In contrast, all three methods failed to demonstrate the presence of virus in organs of immunized guinea pigs after the challenge.

[Infectious bovine rhinotracheitis (VHB-1). II. Physico-chemical properties of strain L-114]. / Rinotraqueítis infecciosa bovina (VHB-1). II: Propiedades físico-químicas de la cepa L-114.

The thermal stability, density gradient and morphological features of a recently isolated L-114 strain of infectious bovine rhinotracheitis virus were determined. Morphology studies showed an enveloped virus with 200 nm of total diameter, a core diameter of 90 nm and an icosaedral-type structure; purified preparations contained both complete and empty viral particles. More than 90% of the infectivity was lost after 15 hours at 37 degrees C; at 56 degrees C, inactivation was much faster, with a 3 log titer reduction, in 24 minutes. Density gradient studies in cesium chloride, carried out with virus concentrated on sucrose gradient, gave an estimated density of 1.25 g/ml for the purified virus, which fits with light herpesviruses.

Rinotraqueitis infecciosa bovina (VHB-1). II: Propiedades fisico-quimicas de la cepa L-114. / Infectious bovine rhinotracheitis (VHB-1). II.Physico-chemical properties of the L-114 strain

Se determino la estabilidad termica, densidad de flotacion y morfologia de la cepa L-114 de virus de la rinotraqueitis infecciosa bovina, aislada recientemente en el pais. La morfologia correspondio a un virus con envoltura, de 200 mm de diametro total y 90 mm de diametro del "core". La estructura de este ultimo fue de tipo icosaedrica, observandose particulas vacias y completas en la preparaciones purificadas de virus. A 37o C, cepa L-114 perdio mas del 90% de su infectividad en 15 hs. y a 56o C se inactivo muy rapidamente, perdiendo 3 log en 24 minutos. La densidad de flotacion en cloruro de cesio, luego de su concentracion por sedimentacion a traves de sacarosa al 47% (p/v), indica que el virus purificado tiene una densidad estimada de 1,25 g/ml, correspondiente a los virus herpes denominados livianos

Rinotraqueítis infecciosa bovina (VHB-1). II: Propiedades físico-químicas de la cepa L-114. / [Infectious bovine rhinotracheitis (VHB-1). II. Physico-chemical properties of strain L-114]

The thermal stability, density gradient and morphological features of a recently isolated L-114 strain of infectious bovine rhinotracheitis virus were determined. Morphology studies showed an enveloped virus with 200 nm of total diameter, a core diameter of 90 nm and an icosaedral-type structure; purified preparations contained both complete and empty viral particles. More than 90

of the infectivity was lost after 15 hours at 37 degrees C; at 56 degrees C, inactivation was much faster, with a 3 log titer reduction, in 24 minutes. Density gradient studies in cesium chloride, carried out with virus concentrated on sucrose gradient, gave an estimated density of 1.25 g/ml for the purified virus, which fits with light herpesviruses.

Rinotraqueitis infecciosa bovina (VHB-1). II: Propiedades fisico-quimicas de la cepa L-114. / Infectious bovine rhinotracheitis (VHB-1). II.Physico-chemical properties of the L-114 strain

Se determino la estabilidad termica, densidad de flotacion y morfologia de la cepa L-114 de virus de la rinotraqueitis infecciosa bovina, aislada recientemente en el pais. La morfologia correspondio a un virus con envoltura, de 200 mm de diametro total y 90 mm de diametro del "core". La estructura de este ultimo fue de tipo icosaedrica, observandose particulas vacias y completas en la preparaciones purificadas de virus. A 37o C, cepa L-114 perdio mas del 90% de su infectividad en 15 hs. y a 56o C se inactivo muy rapidamente, perdiendo 3 log en 24 minutos. La densidad de flotacion en cloruro de cesio, luego de su concentracion por sedimentacion a traves de sacarosa al 47% (p/v), indica que el virus purificado tiene una densidad estimada de 1,25 g/ml, correspondiente a los virus herpes denominados livianos