Establishment of a microplate assay for flow cytometric assessment and it is use for the evaluation of age-related phenotypic changes in canine whole blood leukocytes     

Reis AB  Carneiro CM  Carvalho Md  Teixeira-Carvalho A  Giunchetti RC  Mayrink W  Genaro O  Corrêa-Oliveira R  Martins-Filho OA 《Veterinary immunology and immunopathology》2005,103(3-4):173-185

The effectiveness of flow cytometric assays for canine use is still requiring standardization. Despite several studies using purified mononuclear cells, no methodology or reference ranges are available for immunophenotyping of whole blood leukocytes (WBL). Fresh and pre-fixed WBL were used to identify cell-subsets, (Thy-1(+)/CD5(+)/CD4(+)/CD8(+)/CD21(+) and CD14(+)) and measure MHC-II, CD45RA/CD45RB expression. We described here an efficient method for fast quantification of canine-WBL, using pre-fix in a microplate assay, which allows long-term sample storage prior to phenotyping. Decreased percentage of CD5(+)-T-cells within the lymphocyte-gate and increased percentage of CD21(+)-B-cells were observed in young animals, which led to higher T/B cell ratios in middle-aged dogs. Lower numerical counts of Thy-1(+), CD4(+), CD8(+) and CD21(+) lymphocyte were observed when compared to young animals. In addition, we identified an age-related decline of MHC-II/CD45RA expression by lymphocytes. We proposed an improved method for phenotyping of canine peripheral blood mononuclear cells (PBMC) that has significant use for researchers and veterinary clinicians. The hematological changes of senescence previously identified on PBMC could be adequately reproduced on features identified by whole blood. Furthermore, this study supplies normal range references as baseline standards for clinical purposes, besides specific immunological parameters to monitor canine aging process.  相似文献   

Characterization, In Vitro Culture, and Molecular Analysis of Thecaphora solani, the Causal Agent of Potato Smut     

Andrade O  Muñoz G  Galdames R  Durán P  Honorato R 《Phytopathology》2004,94(8):875-882

ABSTRACT The fungus Thecaphora solani (syn.: Angiosorus solani), the causal agent of potato smut, was cultivated in vitro for the first time. Teliospores obtained from galls of infected potato plants were used to inoculate commonly used solid and liquid media. The teliospores produced two kinds of vegetative tissue depending on the nutrient status of the media. A very slow radial-growing, hyaline, and septate mycelium, as usually seen in most of the in vitro-cultivated filamentous fungi, was obtained in wateragar medium after 30 to 40 days. On the other hand, a white, sponge-like mycelial mass was obtained in HCM + 1% activated charcoal, and on common potato dextrose agar or malt-yeast-peptone solid or liquid media, after 40 to 50 days under lab conditions. The identity among teliospores and the sponge-like mycelial mass was corroborated by DNA fingerprinting and partial sequencing of the large subunit (LSU) rDNA region. The sexual cycle of the pathogen was completed under lab conditions based on the development of teliospores on the sponge-like mycelial mass. The first attempt to reproduce the disease under controlled conditions was successful, inducing a gall in a cv. Desirée potato explant cultivated in vitro inoculated with radial-growing mycelia. Phylogenetic analysis of LSU rDNA data of the genus Thecaphora and other smut fungi confirmed the initial classification of the pathogen as T. solani.  相似文献   

Pathology and Behavioral Effects Associated with Henneguya sp. (Myxozoa: Myxobolidae) Infections of Captive Pacu Piaractus mesopotamicus in Brazil     

M. L. Martins  V. N. Souza  F. R. Moraes    J. R. E. Moraes  A. J. Costa  U. F. Rocha 《Journal of the World Aquaculture Society》1997,28(3):297-300

  相似文献   

Selection and testing of epiphytic yeasts to control anthacnose in post-harvest of papaya fruit     

Guy de Capdeville  Manoel Teixeira Souza Jr.  Jansen Rodrigo Pereira Santos  Simone de Paula Miranda  Alexandre Rodrigues Caetano  Fernando Araripe Gonçalves Torres 《Scientia Horticulturae》2007

Anthracnose, caused by Colletotrichum gloeosporioides, is a major post-harvest disease in papaya fruit. The major objectives of the present work were to isolate, select and test the in vitro and in vivo ability of epiphytic microorganisms, isolated from papaya fruit and leaf surfaces, in controlling anthracnose onset after harvest. A total of 75 bacteria, 67 yeasts and 22 mycelial fungi were isolated. Thirty yeast isolates were able to inhibit the mycelial growth of C. gloeosporioide in vitro and seven of those were used in in vivo assays, resulting in the identification of two very effective isolates. Isolate CEN63, identified molecularly as Cryptococcus magnus, was the most effective in controlling the disease and therefore was studied in more detail. The results of the assays with C. magnus provided evidence that when fruit were treated with the antagonists at concentrations of 10⁷ to 10⁸ cells/ml, as early as 24 h, preferentially 48 h, before inoculation with the pathogen, the development of disease was significantly reduced. C. magnus is a potential antagonist for the development of a commercial product. Additional studies on the modes of action of this yeast isolate, as on its ability to interact with fungicides are being conducted to generate solid basis for the development of an environmentally friendly control agent.  相似文献   

Avermectin resistance of the cattle tick Boophilus microplus in Brazil     

Martins JR  Furlong J 《The Veterinary record》2001,149(2):64

  相似文献   

Toxicity of cypermethrin and diazinon to Haematobia irritans (Diptera: Muscidae) in its American southern range     

Guglielmone AA  Castelli ME  Volpogni MM  Medus PD  Martins JR  Suárez VH  Anziani OS  Mangold AJ 《Veterinary parasitology》2001,101(1):67-73

Synthetic pyrethroids are considered currently less effective than organophosphate insecticides for controlling Haematobia irritans on cattle in its American southern range. To test the validity of this hypothesis, horn fly populations from 95 cattle herds in central-northern Argentina and southern Brazil were evaluated to determine 50% lethal concentrations (LC(50)) to cypermethrin and diazinon, respectively. Horn flies were exposed to filter papers and glass vials treated with cypermethrin and diazinon, respectively, and LC(50) values were determined using probit analysis. Two and 78 fly populations showed LC(50) values indicating susceptibility and resistance to cypermethrin, respectively, in comparison to historical LC(50) values obtained when the horn fly populations were susceptible to cypermethrin. Five fly populations had an undetermined resistance status to cypermethrin. The diazinon tests showed no obvious indications of H. irritans resistance. Thirty nine percent of farmers still rely on pyrethroids to control horn fly populations on cattle which represents a decreasing proportion from a few years ago when horn fly control was carried out using pyrethroids almost exclusively. Organophosphates could provide control for horn flies resistant to pyrethroids; however, the reliance on chemicals alone to control H. irritans is not considered a proper approach for the future.  相似文献   

Erratum to "Pathogenic characteristics of Escherichia coli strain isolated from newborn piglets with diarrhoea in Brazil"     

Martins MF  Martinez-Rossi NM  Ferreira A  Brocchi M  Yano T  Castro AF  Silveira WD 《Veterinary microbiology》2001,78(1):91

  相似文献   

Histopathological and immunohistochemical investigations of the hepatic compartment associated with parasitism and serum biochemical changes in canine visceral leishmaniasis   总被引：1，自引：0，他引：1  

Giunchetti RC  Mayrink W  Carneiro CM  Corrêa-Oliveira R  Martins-Filho OA  Marques MJ  Tafuri WL  Reis AB 《Research in veterinary science》2008,84(2):269-277

The immunopathological evaluation of the hepatic compartment associated with parasitism and biochemical findings are essential for understanding the genesis of hepatomegaly in canine visceral leishmaniasis (CVL). Three clinical groups of dogs naturally infected with Leishmania chagasi [i.e., asymptomatic (AD, n=12), oligosymptomatic (OD, n=12) and symptomatic (SD, n=17)] were assessed and compared with a group of non-infected dogs (NID, n=11). Intense reaction of the Kupffer cells, capsule and portal inflammation, and the presence of intralobular granulomas, were observed in the different clinical groups. Dogs in the SD group presented a higher frequency of parasitism compared with the AD group. Inflammatory alterations were more intense in the SD group and were associated with parasitism. Our results indicated an association between histological liver changes and the progression of biochemical alterations according to progression of clinical forms of CVL, and the direct relationship between clinical symptoms and frequency of hepatic parasitism.  相似文献   

Antigenicity of a whole parasite vaccine as promising candidate against canine leishmaniasis     

Giunchetti RC  Reis AB  da Silveira-Lemos D  Martins-Filho OA  Corrêa-Oliveira R  Bethony J  Vale AM  da Silva Quetz J  Bueno LL  França-Silva JC  Nascimento E  Mayrink W  Fujiwara RT 《Research in veterinary science》2008,85(1):106-112

Human visceral leishmaniasis, one of the most important zoonoses, is caused by the protozoa Leishmania chagasi (syn. L. infantum) and is present as a fatal disease common in South America and Europe where dogs and wild canids are the main reservoirs. A vaccine against visceral leishmaniasis would be an important tool in the control of this disease in dogs. Although the current strategies for vaccination against leishmaniasis are based on the use of recombinant antigens, killed vaccines are still attractive in terms of stability of their biochemical composition and antigenicity, cost, and safety. Here we evaluate the immunogenicity of a whole parasite vaccine as a promising candidate against canine leishmaniasis, demonstrated by cellular reactivity, changes in the cellular profile of the peripheral blood and by the differential production of immunoglobulins. Our results showed that immunization elicited mainly a strong cellular reactivity and increase in T-lymphocytes, particularly the subpopulation CD8(+) that would be related to the control of tissue parasitism. In addition, a higher production of anti-Leishmania total IgG, characterized by mixed isotypes profile (IgG1 and IgG2), was demonstrated.  相似文献   

Evaluation of colostral immunity in swine with commercial anti-leptospira polyvalent whole-bacteria vaccine     

Martins Soto FR  Regina Pinheiro S  Honma Ito F  Maria Moraes Z  Paldes Gonçales A  Santos de Azevedo S  Bernardi F  Rodrigues Camargo S  Arruda Vasconcellos S 《Comparative immunology, microbiology and infectious diseases》2008,31(4):327-335

The intensity and duration of passive immunity against swine leptospirosis were investigated by the microscopic agglutination test and in vitro leptospira growth inhibition. Twenty-one females at first parturition were divided into three groups: Group A (n=08): received two doses with 30 days interval of the commercial anti-leptospira bacterin A. Group B (n=06) received two doses with 30 days interval of the commercial anti-leptospira bacterin B and Group C (n=07) was the control. In all groups the colostrums were collected. Blood collection of piglets was performed in four different ages. Agglutinin antibodies were equally detected in sera and colostrums for serovars Canicola, Grippotyphosa, Copenhageni, Icterohaemorrhagiae and Pomona (Group A) and Canicola, Copenhageni, Icterohaemorrhagiae, Pomona and Hardjo (Group B). Mean neutralizing antibodies titers were low. Passive immunity was low duration.  相似文献