Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin along with marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Hematology of Swiss mice (Mus musculus) of both genders and different ages

PURPOSE: To describe the hematologic values of male and female, young and adult, Swiss mice (Mus musculus). METHODS: Mus musculus (n=14) were randomly selected and separated by gender. The male and female, young and adult animals were sedation to obtain a blood sample, by intracardiac route at 30, 45, 60, 75, 90, 105 and 120 days after birth. RESULTS: The Swiss mouse hemogram values obtained, in relation to total eosinophils, basophils, and number of platelets, there was no statistical differences according to the genders or the age of the animals. Regarding the erythrocyte, hemoglobin and hematocrit values obtained, these were higher in females. The RDW-CD and MPV values were higher in the females than in the males. CONCLUSIONS: Lymphocytes are the predominant cells in the peripheral blood. The collection of 800 µL of blood by intracardiac route, every 15 days, did not affect the health of the animals. Analyses of the blood samples contribute to the experimental models provided by the Central Animal Facility of UFMS and used by professors. .

Hematological changes in mice exposed to biting of the bedbug: Cimex lectularius L. [Hemiptera: Cimicidae]

The studies on hematologic changes in humans or animals as a result of bedbug bites are lacking. This study was undertaken to examine changes in the blood picture of mice [Mus musculus] exposed to Cimex lectularius biting. As compared to the check animals, mice exposed to bedbug bites either once or twice within 7 days showed insignificantly higher WBC's [1.6 and 2.8% increase, respectively] and lower HGB content [0.5 and 0.8% decrease, respectively] and significantly higher PLT's [P<0.01] by 2.2% and 3.0%, respectively. Significantly higher [P<0.01] RBC's counts in mice bitten once than those of normal animals or those exposed to twice bites [5.3 and 5.9% increase, respectively]. Bedbug biting exerts its effects largely upon the differential WBC's. Mice bitten once or twice showed significantly lower number of neutrophils [1.2% and 12.1% decrease, respectively] than those for normal animals. Mice exposed to twice bites showed significantly [P<0.01] higher numbers of lymphocyte [18.8%], monocyte [13.6%], eosinophil [200.0%] and basophil [500%] than those of normal mice

Estudio cariométrico de placentas de ratones con infección aguda por diferentes cepas de Trypanosoma cruzi / Karyometric study of placentas from mice with acute infection by different strains of Trypanosoma cruzi

El objetivo de este trabajo fue evaluar cariometricamente las alteraciones causadas por diferentes cepas de T. cruzi en la placenta del ratón. Ratones hembras de 60 días, grávidas, fueron inoculadas, intraperitonealmente, con 2 x 10(5) tripomastigotes sanguíneos de las cepas colombiana, Y, Solivia o RC del T. cruzi. Fueron observadas claras diferencias en las alteraciones cariométricas de las células trofoblásticas gigantes y de las células trofoblásticas de la zona esponjosa. Los resultados demostraron que las cepas colombiana y RC causan alteraciones tanto en las células trofoblásticas gigantes como en las células del trofoblasto esponjoso, mientras que las cepas Y y Bolivia provocan alteraciones solamente en las células trofoblásticas gigantes. Es posible concluir que cada cepa posee características propias y que, a pesar del tipo similar de transmisión, presenta matices diferenciales en el proceso de la patogénesis placentaria.

The objective of this work was to evaluate karyometrically the alterations caused by different strains of Trypanosoma cruzi in the mouse placenta. Pregnant mice, 60-day old, were intraperitoneally inoculated with 2 x 10(5) bloodstream trypomastigotes of the Colombian, Y, Bolivia or RC strain of T cruzi. There were observed clear differences in the karyometric alterations of the trophoblast giant cells and in the spongiotrophoblast cells. The results demonstrate that the Colombian and RC strains cause alterations both in the trophoblast giant cells and in the spongiotrophoblast cells, whereas the Y and Bolivia strains provoke alterations only in the trophoblast giant cells. It is possible concluding that each strain has its own characteristics and that, in spite of the similar type of transmission, it show differential nuances in the placental pathogenic process.

Study of the genotoxic effect of cyclophosphamide on albino mice bone marrow polychromatic erythrocytes -and the protective effect of captopril

Cyclophosphamide [CYP] is widely used as an antineoplastic and an immunosuppressive drug. However, it has been found to cause DNA damage in normal tissues as well. Captopril [CAP], an angiotensin converting enzyme inhibitor, was reported to have a potential protective effect on the genotoxic effect of CYP possibly through its antioxidant effect. The aim of the present work is to experimentally detect the genotoxic effect of cyclophosphamide using in vivo micronuclei assay in albino mice bone marrow polychromatic erythrocytes and to test the protective effect of captopril on reducing the genotoxicity of CYP. In the present study thirty adult male albino mice were equally divided into six groups. Group I [control group] animals received single physiological saline, group II mice received single injection of captopril [CAP] [50mg/kg], group III animals received single injection of 25mg/kg cyclophosphamide [CYP] dissolved in physiological saline, group IV mice received single injection of 50 mg/kg CYP dissolved in physiological saline, and groups V and VI were the same as group III and IV but CYP injection was preceded by CAP [50mg/kg] injection. The number of micronucleated polychromatic erythrocytes [MNPCEs] was determined in 1000 polychromatic cells from bone marrow smears obtained after sacrificing the animals 24 hrs from exposure to CYP or the control substance. Statistical comparison of the different groups showed that the difference between group I and II was not statistically significant [P=0.106], indicating that CAP does not induce genotoxicity. Whereas, comparing Groups III, IV to group I showed that the difference was statistically significant [P=0.013, 0.00021] It was observed that CYP increased the number of MNPCEs in a dose dependent way. Comparison of groups V and Vito groups III and IV respectively showed a significantly lower number of MNPCEs confirming a protective effect of CAP when administered prior to CYP. The results of the present study confirm a protective role of CAP and support the possibility of administration of captopril prior to cyclophosphamide to ameliorate its genotoxic effect and the possibility to develop secondary cancers

Estudio morfométrico del hígado de ratón en la enfermedad de Chagas experimental / Morphometrical study of the mouse liver in the experimental Chagas' disease

El objetivo de este trabajo fue caracterizar histopatológicamente y morfométricamente las alteraciones del tejido hepático de ratón, durante la fase aguda de la infección por la cepa MORC-2 de Trypanosoma cruzi. Esta cepa mostró acentuado tropismo por el hígado, con numerosos nidos de amastigotes en los cortes examinados. El hígado de los animales infectados estaba constituido por células menores, con citoplasma granuloso. En algunas áreas, los sinusoides estaban congestionados y las células de Kupffer hipertróficas e hiperplásicas. El tejido hepático mostró focos circunscritos de células inflamatorias en áreas de necrosis, sinusoides, en torno de las venas centrolobulillares y de los espacios porta. La vena centrolobulillar estaba dilatada y congestionada, con necrosis focales y ruptura de la pared en algunos campos. Los espacios porta estaban desorganizados, a veces, con intenso infiltrado inflamatorio. En algunas áreas fue posible observar degeneración cística (spongis hepatis). Por todo el tejido hepático se observaron nidos de amastigotes, de tamaño variable, algunos rodeados por infiltrado inflamatorio crónico. En el espacio porta, el volumen relativo de los conductos biliares y vasos sanguíneos, así como la densidad de superficie de las arterias fueron mayores en el grupo infectado.

The objective of this work was to characterize histopatologically and morphometrically the alterations of the mouse liver during the acute infection by the MORC-2 strain of Trypanosoma cruzi. This strain showed marked tropism by the liver, with numerous nests of amastigotes in the examined sections. The liver of the infected animals was constituted by smaller cells, with granular cytoplasm. In some areas, the sinusoids were congested and the Kuppfer cells were hipertrofied and hiperplasic. The hepatic tissue showed circumscribed foci of inflammatory cells into necrotic areas, sinusoids, around the contrilobular veins and the portal spaces. The centrilobular vein was dilated and congested, with focal necrosis and rupture of the wall in some regions. The portal spaces were disorganized, sometimes with intense inflammatory infiltrate. In some areas it was possible to observe cystic degeneration (spongis hepatis). In the hepatic tissue, nests of amastigotes, of variable sizes, were observed, some surrounded by chronic inflammatory infiltrate. In the portal space, the relative volume of the biliary ducts and blood vessels, as well as the surface density of the arteries was greater in the infected group.

Intercellular adhesion molecule-2 [ICAM2] in experimintal trichinosis

The estimation of the serum adhesion molecules [ICAM-2] levels in experimental trichinosis and evaluation of its impact on the course of infection as well as its possible role in the diagnosis of early trichinosis were studied. Serum levels of ICAM-2 correlated significantly with the inflammatory and course sequences of trichinosis in mice and had a similar relation with blood eosinophilia. So, estimation of ICAM-2 serum levels may prove useful in diagnosis of trichinosis recent infections, and in monitoring the prognosis and response to treatment. Investigations of the ICAM-2 role in human trichinosis and applying the findings to improve the diagnostic approaches are highly recommended

Infectivity of cysts of the ME-49 Toxoplasma gondii strain in bovine milk and homemade cheese

Objetivo: Analisar a infecciosidade e a resistência de cistos de T. gondii em leite e queijo fresco caseiro, pela infecção artificial de leite bovino. Métodos: O leite bovino pasteurizado foi infectado artificialmente com 10 cistos/ml de T.gondii cepa ME49 e inoculado em grupos de camundongos, imediatamente ou após ser estocado por 5, 10 e 20 dias a 4oC. Preparou-se queijo fresco caseiro com leite infectado, sendo testado em grupos de camundongos, utilizando a mesma conservação. A infecção foi detectada pela presença de cistos no cérebro dos camundongos desafiados ou testes sorológicos após cinco semanas, também confirmada por Western Blotting e histologia. Resultados: A infecciosidade dos cistos da cepa ME49 de T.gondii foi mantida mesmo quando armazenado no leite até 20 dias de conservação em condições de refrigeração a 4oC. Os cistos resistiram ao processo de fabricação do queijo e eram infectantes após um período de 10 dias nas mesmas condições. Conclusões: Os achados mostraram que o leite e seus derivados podem ser uma importante fonte de contaminação humana pelo T.gondii, reforçando a importância da pasteurização do leite antes de qualquer processamento ou ingestão

Evaluación de un conjugado anti-IgG de ratón-fluoresceína mediante técnicas de inmunofluorescencia indirecta y citometría de flujo / Evaluation of an anti-IgG conjugate of mouse-fluorescin by the indirect immunofluorescent techniques and flow citomerty

Se purificó una inmunoglobulina G de ratón a partir de suero por cromatografía de afinidad en proteína A. Con esta preparación se inmunizaron los conejos cuyos sueros fueron capaces de reconocer al antígeno inyectado mediante inmunodifusión doble. Los anticuerpos fueron precipitados del suero de conejo y purificados mediante cromatografía de intercambio iónico. Esta preparación fue conjugada a isotiocianato de fluorescencia según la tecnología convencional. El conjugado obtenido fue evaluado con las cepas de referencia de virus Parainfluenza 1, 2, 3; Adenovirus; virus sincitial respiratorio y virus influenza A y B por una técnica de inmunofluorescencia indirecta y muestras positivas de VIH mediante citometría de flujo. En ambos casos se utilizaron anticuerpos monoclonales específicos. Se evaluaron muestras clínicas de pacientes con infección respiratoria aguda

Estandarización de la reacción de polimerización en cadena en muestras de sangre de ratones infectados con Trypasonoma cruzi / Standarization of the polymerase chain reaction in blood samples of mice infected with Trypasonoma cruzi

Se estandarizó la reacción de polimerización en cadena (PCR) para el diagnóstico parasitológico de la enfermedad de Chagas. Se determinó condiciones de estandarización de DNA, así como las concentraciones óptimas de dNTPs, partidores, Taq DNA polimerasa y las condiciones de termociclación. Se determinó que el ensayo es capaz de amplificar hasta 100 fg de DNa templado de trypasonoma cruzi, con una sensibilidad diagnóstica de 2x10² parásitos/ml. Los productos de PCR amplificados demostraron ser específicos mediante ensayos de hibridación con una sonda de DNA kinetoplastídico de T. cruzi

Desarrollo del virus de la encefalitis equina venezolana en el tejido nervioso de ratones recién nacidos: ultraestructura e histoquímica / Ultrastructural and histochemical: study on the development of venezuelan equine encephalitis virus in the central nervous system of newborn mice

Se estudio la estructura del tejido nervioso de ratones recién nacidos inoculados con el virus de la EEV. Las células gliales mostraron hichamiento y aumento del glucógeno; las neuronas presentaron alteraciones en la estructura de las mitocondrias y en la organización de los ribosomas; el complejo de Golgi presentó gran desarrollo y tortuosidad de sus cisternas. La actividad de fosfatasa ácida en la cisterna perinuclear y en el retículoendoplasmático granular, sugiere un aumento de producción de esta enzima en las neuronas invadidas por el virus de la EEV. Se estudió la localización de las partículas virales en relación con la viremia y diseminación de la infección, destacándose la presencia de partículas virales en la luz de los vasos sanguíneos y alrededor de las vainas de mielina de algunos axones. El desarrollo de las partículas virales se comparó con las observaciones hechas en otras arbovirosis, proponiéndose que los precursores virales se originen en masas de viroplasma, desde donde migran a través del citoplasma, hacia las cisternas y vacuolas del complejo de Golgi o hacia la membrana plasmática, transformándose en partículas virales maduras por gemación

GABA metabolism in venezuelan equine encephalomyelitis virus infection: preliminary communication / Metabolismo de GABA en la infección producida por el virus de la encefalitis equina venezolana

Mice infected with venezuelan equine encephalomyelitis virus showed a significant decrease in GABA content of cerebral hemispheres. Activity of the enzyme which synthesizes GABA, glutamic acid decarboxylase, is also reduced in whole cerebral hemispheres, caudate nucleus, and frontal cortex of infected animals, as compared to values obtained from the same regions of control mice

Haemopoietic stem cells during development of mouse embryo.

Haemopoiesis in mammals takes place in yolk-sac and in mouse it can be detected on the 7th day of gestation. Erythropoietin (EPO) responsive cells can be detected from 7th day onwards. However, the cells committed to the myeloid lineage which can respond to the haemopoietic growth factor (viz. granulocyte macrophage colony stimulating factor; GM-CSF) can be demonstrated only on 10th day of gestation. At the same time, the 12-day spleen colony forming cells i.e. the late colony forming unit spleen (CFU-s) which are multipotent stem cells can also be detected. Data suggest that the stem cells seen in the embryo from 7-10 days of gestation may be a primitive population confined only to the yolk-sac. Liver haemopoiesis which begins in the liver of 13-day embryos is due to primitive haemopoietic pluripotent stem cells, arising de novo in the embryo and not in the yolk-sac, since no primitive pluripotent stem cells capable of repopulating lethally irradiated bone-marrow can be detected in the yolk-sac.

Comparative study of different methods used to isolated Trypanosoma cruzi from defibrinated blood of irradiated mice

Different methods are being used for the isolation and purification of Trypanosoma cruzi blood forms from infected vertebrate hosts. In this study we compare four of these methods (differential centrifugation, Ficoll-Hypaque, Histopaque 1077 and metrizamide) in terms of parasite recovery rates, contamination with cells, duration of the process and role of host irradiation. male albino Swins mice irradiated in a Gamma Cell 220 (500 rads) were inoculated with CL and VL-10 T. cruzi strains and bled at the peak of parasitemia. Infected defibrinated blood was then used for the isolation. Although all methods permitted the recovery of viable trypomastigotes, the best results were obtained with Ficoll-Hypaque and Histopaque 1077. Recovery rates ranged between 71% to 88% and parasite-enriched preparations were obtained in approximately 75 min. irradiation and blood defibrination drastically reduced platelet and leukocyte contamination of the preparations

Isotypic distribution of antibody responses in lines of mice selected for high or low immunoresponsiveness

The isotype distribution of antibody (Ab) responses to Salmonella antigens (Ag) was investigated in high (H) and low (L) Ab responder lines of mice from Selections III and carried out for responsiveness to flagellar (f) and somatic (s) Ag, respectively. Primary immuniztion resulted in higher Ab titers of all isotypes in response to both Ag in H mice fro m both selections and was confirmed after booster injections. The interline difference (H-L) in response to the distinct isotypes ranged from 3.0 to 7.0 log2 to Ag f in Selection III and from 2.0 to 5.1 log2 to Ag s in Selection IV. Comparison of isotype production to 3 Ag in Selections I,II,III and IV demonstrated that: 1) the highest responses in all mice are those against the selection Ag, 2) the isotypic pattern depends on both the Ag injected and the host's genetic constitution, and 3) the presence or lack of a multispecific effect is not due to isotype-restricted regulation