Papers by Víctor Tenorio-gutiérrez
AN ISOLATED PROTEIN FRACTION OF VENOM OF AGKISTRODON PISCIVORUS WITH HEMAGGLUTINANT PROPERTY WAS ... more AN ISOLATED PROTEIN FRACTION OF VENOM OF AGKISTRODON PISCIVORUS WITH HEMAGGLUTINANT PROPERTY WAS EVALUATED IN ITS CAPACITY TO INHIBIT IN VITRO THE BOVINE PARAINFL UENZA-3 (PI3) VIRUS, EITHER BY ITS CAPACITY TO BLOCK CELL RECEPTORS OR ITS ADHERENCE TO THE HEMAGGLUTININ VIRAL SPICULES. FRACTION AL27 ACTING AS A RECEPTOR BLOCK, AT A CONCENTRATION 1.062 A“G/ML, INHIBITED ANY DAMAGE OR DESTRUCTION OF MADIN DARBY BOVINE KIDNEY (MDBK) CELL CULTURES BY THE PI-3 VIRUS (TITER 105.6 TCID50%), MAINTAINING A CELL VIABILITY BETWEEN 69.43 AND 84.86%. THE ADHERENCE CAPACITY OF AL27 TO VIRAL HEMAGGLUTININ SPICULES OF PI3 VIRUS WAS DETERMINED BY INCUBATING BOTH REACTANTS 1:1 (1 ML AL27 AT CONCENTRATION 1.062 A“G/ML 1 ML PI-3 VIRUS TITER 105.6 DICC50%) AT 37°C FOR 1 HOUR, AND A PROBABLE ADHERENCE TO THE VIRAL HEMAGGLUTININ SPICULES WAS DETECTED BY ELECTRON MICROSCOPE. THE INACTIVATION OF PI-3 VIRUS BY AL27 WAS DEDUCED BY THE REDUCTION OF THE VIRAL TITER FROM 105.6 TO 102.0 TCID50%. THE APPARENT IDENTI...
Revista Mexicana de Ciencias Pecuarias
Se evaluó la virulencia de cepas de Listeria monocytogenes todas de serotipo 4b procedentes de ca... more Se evaluó la virulencia de cepas de Listeria monocytogenes todas de serotipo 4b procedentes de cabras y susderivados. Se observaron niveles de virulencia variables cuando se comparó la virulencia relativa (porcentaje deletalidad) en ratones BALB/c inoculados vía intravenosa o intragástrica y su capacidad para infectar macrófagos J774A.1, y células epiteliales Caco-2. Dos cepas obtenidas de alimento de cabras produjeron 100 % de letalidad por ambas vías de inoculación y no mostraron diferencia significativa con la cepa testigo (P>0.05) respecto al porcentajede invasión y a los parámetros de la cinética de crecimiento cuadrática observada en ambas líneas celulares. Si bien todas las cepas lograron invadir las células Caco-2, solamente algunas consiguieron invadir el bazo después de la inoculación por vía intragástrica. Las dos cepas provenientes de alimento de cabras fueron las más virulentas, representando un riesgo para la salud humana y animal, ya que pueden ser diseminadas en e...
Pesquisa Veterinária Brasileira, 2016
RESUMO.-[Distribuição de linfócitos, células carreadoras de imunoglobulinas, macrófagos e células... more RESUMO.-[Distribuição de linfócitos, células carreadoras de imunoglobulinas, macrófagos e células dendríticas em glândulas sexuais acessórias de carneiros infectados experimentalmente com Actinobacillus seminis.] A distribuição das células envolvidas na resposta imune em glândulas sexuais acessórias de carneiros expe
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire
Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes protease... more Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes proteases that degrade porcine hemoglobin and IgA in vitro. To further characterize A. pleuropneumoniae proteases, we constructed a genomic library expressed in Escherichia coli DH5ox, and selected a clone that showed proteolytic activity. The recombinant plasmid carries an 800-base pair A. pleuropneumoniae gene sequence that codes for a 24-kDa polypeptide. A 350-base pair PstI fragment from the sequence hybridized at high stringency with DNA from 12 serotypes of A. pleuropneumoniae, but not with DNA from Actinobacillus suis, Haemophilus parasuis, Pasteurella haemolytica, Pasteurella multocida A or D, or E. coli DH5cx, thus showing specificity for A. pleuropneumoniae. The expressed polypeptide was recognized as an antigen by convalescent-phase pig sera. Furthermore, a polyclonal antiserum developed against the purified polypeptide recognized an A. pleuropneumoniae oligomeric protein in both crude-extract and cell-free culture media. This recombinant polypeptide cleaved azocoll, gelatin, and actin. Inhibition of the proteolytic activity by diethylpyrocarbonate suggests that this polypeptide is a zinc metalloprotease. Resume Actinobacillus pleuropneumoniae est l'agent etiologique de la pleuropneumonie porcine. Cette bacterie secrete des proteases qui degradent l'hemoglobine et les IgA d'origine porcine in vitro. Afin de mieux caracteriser les proteases d'A. pleuropneumoniae, une librairie
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire
Pseudomonas pseudomallei is the causative agent of melioidosis, a glanders-like disease of humans... more Pseudomonas pseudomallei is the causative agent of melioidosis, a glanders-like disease of humans and animals. The pathogenesis of melioidosis is not well understood, and the role of various extracellular enzymes produced by P. pseudomallei in the development of this disease is not known. The present studies were designed to purify and characterize an extracellular protease produced by P. pseudomallei isolates and to test the hypothesis that this protease may play a role in melioidosis. The protease was present in culture supernatants as an enzyme with a molecular weight of 36 000 that was optimally active at 60 °C and at pH 8.0. The P. pseudomallei protease was shown to be a metalloenzyme requiring iron for maximal activity, and activity was inhibited in the presence of ethylenediaminetetraacetic acid (150 mM). Antibodies directed against an alkaline protease produced by Pseudomonas aeruginosa cross-reacted with the P. pseudomallei protease. These data indicate that the P. pseudomallei protease belongs to the family of alkaline proteases sensitive to metal chelators. Purified P. pseudomallei protease was capable of digesting a variety of eucaryotic protein substrates including immunoglobulins. A P. pseudomallei strain deficient in protease production was shown to be less virulent than the parental strain in an animal model of lung infection. These data suggest that this protease may be a significant pathogenic determinant in infections caused by P. pseudomallei.Key words: protein, pathogenesis, substrates, disease.
Tecnica Pecuaria en Mexico
La salmonelosis producida por Salmonella enteritidis (SE), es una enfermedad que afecta a las ave... more La salmonelosis producida por Salmonella enteritidis (SE), es una enfermedad que afecta a las aves y a los mamíferos, incluyendo al hombre, por lo que se le considera una zoonosis. El propósito del presente estudio fue relacionar la presencia de plásmidos y la resistencia antimicrobiana en cepas de SE de origen aviar. Se trabajaron 50 cepas de SE, 21 del fagotipo 4 y 29 del fagotipo 8. Para realizar la prueba de resistencia antimicrobiana se utilizó la técnica de difusión en placa con un Kit comercial, siguiendo la técnica de Kirby-Bauer. Para la extracción del plásmido se utilizó el método lisis-alcalina. Se encontró una resistencia de un 100 % a amikacina, gentamicina, netilmicina y nitrofurantoína en cepas que contenían desde uno hasta cinco plásmidos, por lo que no es posible relacionar la resistencia encontrada en algunas cepas debido a la presencia de plásmidos. Todas las cepas presentaron un plásmido común de 12 Kpb. En todas las cepas del fagotipo 4 se observó la presencia d...
The AL27 crotalic fraction is a lectin recognized by cell receptors and then introduced into the ... more The AL27 crotalic fraction is a lectin recognized by cell receptors and then introduced into the cytoplasm. In the present research a new transfection promoter agent for a non viral expression vector was used, using as a model a plasmid expressing the gene that codifies for the G glycoprotein of the rabies virus. The objective was to evaluate the ability of AL27 to increase the rabies plasmid transfection in in vitro and in vivo conditions. The AL27 fraction was isolated from the venom of Agkistrodon piscivorus by HPLC-RP. AL27 maintained its recognition by sialic acid receptors up to a titer of 64 HU. The pC38 plasmid was selected from a panel of 18 clones constructed by inserting the gene that codifies for glycoprotein G of rabies strain HQIMSS99 into the commercial pCI-neo expression vector. In vitro results showed that pC38, with or without AL27, transfected CHO cells under the calcium-phosphate system and using Confocal Microscopy (CM) increased fluorescence (+++) was observed ...
Actinobacillus pleuropneumoniae serotype 1 adhered to immobilized swine-lung collagen. Bacteria b... more Actinobacillus pleuropneumoniae serotype 1 adhered to immobilized swine-lung collagen. Bacteria bound to collagen type I, III, IV and V. At 5 min incubation, 30 % of bacteria adhered to collagen, reaching saturation in around 90 min. Treatment of bacteria with divalent-metal chelators diminished their attachment to collagen, and Ca 2+ but not Mg 2+ increased it, suggesting Ca 2+ dependence for adherence. Proteolytic enzymes drastically reduced bacterial adherence to collagen, showing that binding involved bacterial surface proteins. Porcine fibrinogen, haemoglobin and gelatin partially reduced collagen adhesion. A 60 kDa outer-membrane protein of A. pleuropneumoniae recognized the swine collagens by overlay. This membrane protein was apparently involved in adhesion to collagen and fibrinogen, but not to fibronectin and laminin. Antibodies against the 60 kDa protein inhibited the adhesion to collagen by 70 %, whereas pig convalescent-phase antibodies inhibited it by only 40 %. Serotypes 1 and 7 were the most adherent to pig collagen (taken as 100 %); serotypes 6 and 11 were the lowest (~50 %), and neither showed the 60 kDa adhesin to biotinylated collagens. By negative staining, cells were observed initially to associate with collagen fibres in a polar manner, and the adhesin was detected on the bacterial surface. The results suggest that swine-lung collagen is an important target for A. pleuropneumoniae colonization and spreading, and that the attachment to this protein could play a relevant role in pathogenesis.
In experimentally infected rams with Actiobacillus seminis, pathological changes (reproductive or... more In experimentally infected rams with Actiobacillus seminis, pathological changes (reproductive organs) and antibodies (IgM, IgG and IgA) in serum, seminal plasma and smegma were evaluated. Twelve 1-year old rams were infected via intraurethal IU group (n = 4) and intraepididymal IE group (n = 4) with A. seminis; four animals were kept as Controls (CON) and inoculated both ways (intraepididymal and intraurethal) with saline solution. Blood, seminal plasma and smegma were collected, during 9 weeks: 4 before and 5 after inoculation. Bacteriological analysis of semen was performed. Animals were euthanized 35 days postinoculation; reproductive organs were collected for bacteriological and histopathological analysis. IE animals developed epididymitis. Ampullae and seminal vesicles in the IE and IU showed interstitial lymphoplasmacytic infiltrates. Thus, the bacteria were not re-isolated. Two controls developed epididymal granuloma. A. seminis was re-isolated in most of the semen samples a...
A routine andrology and semen examination was carried out on a group of 12, one-year old, Pelibue... more A routine andrology and semen examination was carried out on a group of 12, one-year old, Pelibuey breed rams, but one individual had persisting leukocytospermia. No clinical alterations of the reproductive system were found in any of the rams. At slaughter of the problem animal, the seminal vesicles were found to be markedly lobulated and larger than normal, as were the vas deferens ampoules. When dissected, both structures contained creamy white yellowish material. Histopathology demonstrated ampullitis and seminal vesiculitis. A Gram-negative pleomorphic bacillus was isolated from both of the organs, which was characterized through the API 20E system and PCR. It was found to behave similar to the reference strain for Actinobacillus seminis ATCC 15768.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, 2006
Actinobacillus seminis is a gram-negative bacterium of the Pasteurellaceae family that is involve... more Actinobacillus seminis is a gram-negative bacterium of the Pasteurellaceae family that is involved in ovine epididymitis. Looking for a protein specific to this species, we determined the protein profile of subcellular fractions of A. seminis (American Type Culture Collection number 15768): proteins from the outer membrane (OMPs), inner membrane (IMPs), and cytoplasm (CPs). These profiles provide the first data, to our knowledge, regarding subcellular fractions of A. seminis. In the OMP fraction, we identified a protein with a molecular mass of 75 kDa that proved to be immunogenic and apparently specific for A. seminis. This conclusion was based on the reaction of hyperimmune serum of rabbits inoculated with whole cells of A. seminis that was tested against sonicated complete cells of reference strains and field isolates of Brucella ovis, Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni. No protein of these bacteria cross-reacted with the 75-kDa protein of A. sem...
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, 2000
Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes protease... more Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes proteases that degrade porcine hemoglobin and IgA in vitro. To further characterize A. pleuropneumoniae proteases, we constructed a genomic library expressed in Escherichia coli DH5alpha, and selected a clone that showed proteolytic activity. The recombinant plasmid carries an 800-base pair A. pleuropneumoniae gene sequence that.codes for a 24-kDa polypeptide. A 350-base pair PstI fragment from the sequence hybridized at high stringency with DNA from 12 serotypes of A. pleuropneumoniae, but not with DNA from Actinobacillus suis, Haemophilus parasuis, Pasteurella haemolytica, Pasteurella multocida A or D, or E. coli DH5alpha, thus showing specificity for A. pleuropneumoniae. The expressed polypeptide was recognized as an antigen by convalescent-phase pig sera. Furthermore, a polyclonal antiserum developed against the purified polypeptide recognized an A. pleuropneumoniae oligomeric protein in bot...
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, 1994
It was found that 48 hour cultures of Actinobacillus pleuropneumoniae secreted proteases into the... more It was found that 48 hour cultures of Actinobacillus pleuropneumoniae secreted proteases into the medium. Electrophoresis in polyacrylamide gels (10%) copolymerized with porcine gelatin (0.1%), of the 70% (NH4)2SO4 precipitate from the culture supernatants, displayed protease activities of different molecular weights: > 200, 200, 90, 80, 70 and 50 kDa. They had activity over a broad range of pHs (4-8), with an optimal pH of 6-7. All were inhibited by 10 mM EDTA, and reactivated by 10 mM calcium. They were stable at -20 degrees C for more than a month. The proteases also degraded porcine IgA and porcine, human, and bovine hemoglobin, although they appeared to be less active against the hemoglobins. The IgA was totally cleaved in 48 h, using supernatants concentrated with polyvinyl pyrrolidone or the 70% (NH4)2SO4. Extracellular proteases could play a role in virulence.
Frontiers in Cellular and Infection Microbiology, 2012
The virB locus, which encodes the type IV secretion system, is a major component of virulence in ... more The virB locus, which encodes the type IV secretion system, is a major component of virulence in Brucella. A non-polar virB10 mutant and a virB11 deletion mutant were constructed in Brucella canis. In the mouse model, both mutants were cleared at day 21 post-infection, indicating reduced virulence in mice. After challenging with wild-type B. canis, the amounts of CFU recovered at day 15 were significantly lower in the group previously vaccinated with the virB10 mutant. Levels of IgG1, IgG2a, IgG2b, and IgM, the induction of the cytokines IL-2, IL-4, IL-10, and the production of IFN-γ were measured in lymphocyte cultures. All strains elicited similar levels of different antibody isotype profiles, and no significant differences were detected (P < 0.05).The wild-type strain induced a rapid and strong INF-γ response at 24 h, while both mutants induced mild INF-γ responses at 24 h, which remained constant over the course of sampling. Our results suggest that the virB mutants elicit a protective immunity and may be considered as candidates for studies to be conducted in dogs against canine brucellosis.
Microbiology, 2004
Actinobacillus pleuropneumoniae serotype 1 adhered to immobilized swine-lung collagen. Bacteria b... more Actinobacillus pleuropneumoniae serotype 1 adhered to immobilized swine-lung collagen. Bacteria bound to collagen type I, III, IV and V. At 5 min incubation, 30 % of bacteria adhered to collagen, reaching saturation in around 90 min. Treatment of bacteria with divalent-metal chelators diminished their attachment to collagen, and Ca2+ but not Mg2+ increased it, suggesting Ca2+ dependence for adherence. Proteolytic enzymes drastically reduced bacterial adherence to collagen, showing that binding involved bacterial surface proteins. Porcine fibrinogen, haemoglobin and gelatin partially reduced collagen adhesion. A 60 kDa outer-membrane protein of A. pleuropneumoniae recognized the swine collagens by overlay. This membrane protein was apparently involved in adhesion to collagen and fibrinogen, but not to fibronectin and laminin. Antibodies against the 60 kDa protein inhibited the adhesion to collagen by 70 %, whereas pig convalescent-phase antibodies inhibited it by only 40 %. Serotypes...
Revista Mexicana de …, 2006
Con el objetivo de inducir la infección experimental de A. seminis, se utilizaron 18 corderos de ... more Con el objetivo de inducir la infección experimental de A. seminis, se utilizaron 18 corderos de seis meses de edad: 4 testigos negativos, otros 3 se inocularon con A. seminis vía intraepididimal (IE) y 11 fueron inoculados por vía intrauretral (IU) como sigue: cuatro recibieron factor liberador de gonadotropinas (GnRH) cinco días previos al desafío, cuatro recibieron etilenglicol por vía IE 24 h previas al desafío y tres no recibieron tratamiento previo. De los cuatro testigos negativos, dos recibieron GnRH y dos etilenglicol. Se tomaron muestras semanales de suero para inmunodifusión, y de semen para bacteriología. Se sacrificaron a los 35 días posinoculación, obteniéndose testículos, epidídimos y glándulas anexas, para estudios bacteriológicos e histopatológicos. Del grupo tratado con GnRH, se recuperó la bacteria del semen pero no del aparato reproductor, aunque se observaron lesiones en 3 de 4 animales. Del grupo estimulado con etilenglicol, en uno se aisló A. seminis de ámpulas del deferente y de vesícula seminal. De los desafiados sin tratamiento previo, en 2 de 3 se aisló la bacteria del semen. De los tres inoculados IE, en uno se aisló del lugar de inoculación y en dos se aisló de una de las glándulas anexas. En la serología de los desafiados por vía IU, 2 de 11 resultaron positivos; de los 3 inoculados IE, 2 resultaron positivos. Las observaciones histopatológicas correspondieron a lesiones típicas de epididimitis con infiltrados inflamatorios en glándulas anexas. Los resultados sugieren que las glándulas anexas pueden ser un reservorio de A.seminis.
Veterinaria …, 2009
Detección de secuencias nucleotídicas de Mycobacterium bovis a partir de adn de moco nasal de cap... more Detección de secuencias nucleotídicas de Mycobacterium bovis a partir de adn de moco nasal de caprinos inoculados experimentalmente Detection of Mycobacterium bovis nucleotide sequences from nasal mucus of experimentally inoculated goats Recibido el 5 de julio de 2005 y aceptado el 25 de enero de 2006.
Journal of Animal and …, 2007
Cynthia Gonzalez-Ruiz, Victor Tenorio-Gutierrez, Francisco Trigo-Tavera et al. JOURNAL OF ANIMAL ... more Cynthia Gonzalez-Ruiz, Victor Tenorio-Gutierrez, Francisco Trigo-Tavera et al. JOURNAL OF ANIMAL AND VETERINARY ADVANCES, Vol. 6, No. 10. (OCT 2007). Mannheimia haemolytica is a commensal of cattle, sheep and ...
Avian Pathology, 2005
This article may be used for research, teaching, and private study purposes. Any substantial or s... more This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae, and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand, or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material.
Revista Mexicana de …, 2005
Es reconocido que Salmonella Enteritidis, es uno de los serotipos causantes de infección en aves ... more Es reconocido que Salmonella Enteritidis, es uno de los serotipos causantes de infección en aves y en diferentes especies animales, siendo una zoonosis causante de problemas entéricos en el humano. Se ha considerado al consumo de huevo contaminado como la principal vía de infección para el hombre. El objetivo del trabajo fue determinar la posible presencia de Salmonella Enteritidis en el huevo para consumo humano en la ciudad de México. Se muestrearon 400 huevos de 10 marcas comerciales. Se obtuvieron 131 aislamientos considerados en 12 diferentes géneros bacterianos: Acinetobacter sp., Alcaligenes sp., Bacillus sp., Branhamella sp., Edwardsiella sp., Hafnia sp., Klebsiella sp., Serratia sp., Shigella sp., Staphylococcus sp., Yersinia sp. y obteniendo una cepa de Salmonella Enteritidis, la cual se clasificó como no tipificable. Veinte aislamientos no pudieron ser clasificados. Se obtuvo un huevo contaminado por Salmonella Enteritidis, el cual en porcentaje del muestreo representó el 0.25 %; sin embargo se encontraron además 11 géneros bacterianos contaminantes que pudieran representar un riesgo para la producción avícola y la salud pública.
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Papers by Víctor Tenorio-gutiérrez