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91.
In order to determine the occurrence, serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) strains, 153 fecal samples of cattle randomly selected from six dairy farms in Sao Paulo State, Brazil, were examined for Shiga toxin (Stx) production by the Vero cell assay. Feces were directly streaked onto MacConkey Sorbitol Agar and incubated at 37 degrees C overnight. Sorbitol-negative colonies (maximum 20) and up to 10 sorbitol-positive colonies from each plate were subcultured onto presumptive diagnostic medium IAL. Sorbitol-negative isolates were screened with O157 antiserum for identification of O157:H7 E. coli. Isolates presenting cytotoxic activity were submitted to colony hybridization assays with specific DNA probes for stx1, stx2, eae, Ehly and astA genes. The isolation rate of STEC ranged from 3.8 to 84.6% depending on the farm analysed. STEC was identified in 25.5% of the animals, and most of them (64.1%) carried a single STEC serotype. A total of 202 STEC isolates were recovered from the animals, and except for the 2 O157:H7 isolates all the others expressed cytotoxic activity. The great majority of the STEC isolates carried both stx1 and stx2 genes (114/202, 56.4%) or stx2 (82/202, 40.6%); and whereas the Ehly sequence occurred in most of them (88%) eae was only observed in O157:H7 and O111:HNM isolates. Serotypes O113:H21, O178:H19 and O79:H14 were the most frequent STEC serotypes identified and widely distributed among animals from different farms, while others such as O77:H18, O88:H25 and O98:H17 occurred only in particular farms. This is the first report on the occurrence of STEC in dairy cattle in Sao Paulo State, and the results point to substantial differences in rate of isolation, serotypes and genetic profile of STEC that has been previously described among beef cattle in our community. Moreover, to our knowledge O79:H14 and O98:H17 represent new STEC serotypes, while O178:H19 has only been recently reported in Spain.  相似文献   
92.
OBJECTIVES: To evaluate the role of interleukin (IL)-10 in the inability of monocyte-derived bovine macrophages to kill Mycobacterium avium subsp paratuberculosis organisms in vitro. SAMPLE POPULATION: Monocytes were obtained from healthy adult Holstein dairy cows that had negative results when tested for infection with M avium subsp paratuberculosis. PROCEDURE: Monocyte-derived macrophages were incubated with M avium subsp paratuberculosis for 2, 6, 24, 72, or 96 hours with or without addition of saturating concentrations of a goat anti-human IL-10 that has been documented to neutralize bovine IL-10 activity. Variables assessed included ingestion and killing of M avium subsp paratuberculosis; expression of tumor necrosis factor (TNF)-alpha, IL-12, IL-8, major histocompatability (MHC) class II, vacuolar H+ ATPase, and B cell CLL/lymphoma 2 (BCL-2); production of nitric oxide; acidification of phagosomes; and apoptosis of macrophages. RESULTS: Neutralization of IL-10 enabled macrophages to kill 57% of M avium subsp paratuberculosis organisms within 96 hours. It also resulted in an increase in expression of TNF-alpha, IL-12, IL-8, MHC class II, and vacuolar H+ ATPase; decrease in expression of BCL-2; increase in acidification of phagosomes; apoptosis of macrophages; and production of nitric oxide. CONCLUSIONS AND CLINICAL RELEVANCE: The capacity of M avium subsp paratuberculosis to induce IL-10 expression may be a major determinant of virulence for this organism.  相似文献   
93.
An enteric infection in cockatoos associated with a 30 nm diameter enterovirus-like agent seen in faeces and intestinal epithelial cells is described. The disease is characterised by intractable, profuse, mucoid diarrhoea, weight loss, dehydration and death. Lesions in the intestine consist of villous atrophy, villous fusion, enterocyte hyperplasia and, in some cases, chronic inflammation. Affected birds so far examined have concurrent psittacine beak and feather disease.  相似文献   
94.
Growth and photochemical response of triazine-susceptible and -resistant rutabaga ‘Laurentian’ genotypes to cyanazine (chloro-s-triazine) and metribuzin (methylthio-as-triazine) were evaluated. The young seedlings of the susceptible rutabaga were killed when cyanazine or metribuzin were applied either pre- or post-emergence. The triazine-resistant rutabaga, however, displayed a differential response to these herbicides. Metribuzin applied pre-emergence killed the seedlings at 0.4 kg ha?1, and at 0.2 kg ha?1 the growth was severely affected. Cyanazine even at higher rates applied pre- or post-emergence failed to inhibit growth in these plants.Chlorophyll fluorescence in leaf sections at the cotyledonary, 2-leaf and 4-leaf stages in the susceptible plants increased by 130 and 172% in response to 10?5 M cyanazine and metribuzin, respectively, suggesting that Photosystem II reactions in these plants were severely impaired. In resistant plants, there was little or no leaf chlorophyll fluorescence (LCF) increase at this herbicide concentration. However, at 10?4 M cyanazine and metribuzin, the LCF in resistant plants increased significantly and the increase was greater in response to metribuzin (86%) than to cyanazine (51%). This indicates that with these triazine-resistant genotypes, metribuzin is relatively more potent and at high rates is capable of interfering with photosynthesis.  相似文献   
95.
The prevalence of anti-Toxoplasma gondii antibodies was evaluated by the indirect immunofluorescent-antibody test in serum of 57 wild canids from three different species: Lycalopex gymnocercus, Cerdocyon thous and Dusicyon vetulus from the northeast, southeast and southern regions of Brazil. The prevalence was 35.1%, with 20 of the 57 canids demonstrating antibodies anti-T. gondii at dilutions of 1:16 in 2, 1:32 in 4, 1:64 in 2, 1:128 in 2, 1:256 in 6, 1:512 in 2 and 1:2048 in 2 animals. None of the D. vetulus were positive. Among the L. gymnocercus 11 (91.7%) of the 12 samples were positive and among C. thous 9 (60%) of the 15 had antibodies anti-T. gondii.  相似文献   
96.
Ehrlichia canis is the causative agent of canine monocytic ehrlichiosis. In order to evaluate platelet counts as a screening test for E. canis in an endemic area, 217 whole blood samples from dogs were divided into three groups: 71 non-thrombocytopenic samples (group A, platelet counts greater than 200 000/mL) and 146 thrombocytopenic samples (less than 200 000/mL). The thrombocytopenic group was further divided into 62 with platelet counts between 100 000-200 000/mL (Group B) and 84 samples with less than 100 000 platelets/mL (Group C). All samples were examined for the presence of a segment of the Ehrlichia canis 16S rRNA gene using a nested polymerase chain reaction. Sixty-seven of the 217 samples (30.9%) were positive for the presence of the E. canis 16S rRNA gene; 53 (63.1%) of the group C samples and 13 (21%) of group B. Only one (1.4%) of the non-thrombocytopenic samples (Group A) was positive. These data support the concept that platelet counts may be a good screening test for canine monocytic ehrlichiosis, and that the magnitude of thrombocytopenia may increase the reliability of diagnosis.  相似文献   
97.
OBJECTIVE: To evaluate activation of Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) pathway in bovine monocytes after incubation with Mycobacterium avium subsp paratuberculosis (Mptb) organisms. SAMPLE POPULATION: Bovine monocytes obtained from 4 healthy adult Holstein dairy cows. PROCEDURES: Bovine monocytes were incubated with Mptb organisms with or without a specific inhibitor of the JNK/SAPK pathway (SP600125) for 2, 6, 24, or 72 hours. Expression of interleukin (IL)-1beta, IL-10, IL-12, IL-18; transforming growth factor-beta (TGF-beta); and tumor necrosis factor-alpha (TNF-alpha) and the capacity of Mptb-infected monocytes to acidify phagosomes and kill Mptb organisms were evaluated. Phosphorylation status of JNK/SAPK was evaluated at 10, 30, and 60 minutes after Mptb incubation. RESULTS: Compared with uninfected control monocytes, Mptb-infected monocytes had increased expression of IL-10 at 2 and 6 hours after incubation and had increased expression of TNF-alpha, IL-1beta, IL-18, and TGF-beta at 2, 4, and 6 hours. Additionally, Mptb-infected monocytes had increased expression of IL-12 at 6 and 24 hours. Addition of SP600125 (specific chemical inhibitor of JNK/SAPK) resulted in a decrease in TNF-alpha expression at 2, 6, and 24 hours, compared with untreated Mptb-infected cells. Addition of SP600125 resulted in a decrease in TGF-beta expression at 24 hours and an increase in IL-18 expression at 6 hours. Addition of SP600125 failed to alter phagosome acidification but did enhance the capacity of monocytes to kill Mptb organisms. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of JNK/SAPK may be an important mechanism used by Mptb to regulate cytokine expression in bovine monocytes for survival and to alter inflammatory and immune responses.  相似文献   
98.
The intestinal protozoan parasite Giardia duodenalis (Lambl, 1859) Kofoid & Christiansen, 1915 [syn. Giardia intestinalis and Giardia lamblia] has emerged as a widespread enteric pathogen in humans and domestic animals. In recent years, G. duodenalis has been found in cattle worldwide and longitudinal studies have reported cumulative prevalence of 100% in some herds. In the present study, we determined the prevalence and genetic characterisation of G. duodenalis in 200 dairy cattle from 10 dairy farms in S?o Paulo state, Brazil. All faecal specimens were screened for the presence of G. duodenalis using microscopy examination, enzyme immunoassay (EIA) and polymerase chain reaction (PCR). DNA was extracted from faecal samples and G. duodenalis were identified by amplification of the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes followed by restriction fragment length polymorphism (RFLP) or sequencing analysis. Giardia was identified in eight farm locations (80% prevalence). Overall, 15/200 (7.5%) animals were positive for infection, only one of which was a cow. Giardia duodenalis genotype E was present in 14 of the animals tested. Zoonotic genotype AI was present in one positive sample. Genotype E and genotype A represented 93% and 7% of G. duodenalis infections, respectively. This study demonstrates that G. duodenalis infection was prevalent in dairy calves in S?o Paulo state and that the non-zoonotic genotype E predominates in cattle in this region. Nevertheless, calves naturally infected in Brazil can shed Giardia cysts that can potentially infect humans, and thus, they may represent a public health risk.  相似文献   
99.
Colletotrichum lindemuthianum, causal agent of anthracnose in the common bean, has wide genetic variability. Differential bean cultivars and morphological and physiological characteristics were used to analyze 74 isolates of C. lindemuthianum collected in two counties in the state of Minas Gerais, Brazil. Six different races were found, with a predominance of race 65 at both locations. Isolates were classified according to their sensitivities to the fungicide thiophanate-methyl, normally used in the control of common bean anthracnose. In all, ≈10% of isolates were resistant to the fungicide in vitro. Characteristics such as indexes of mycelia growth rate, colony diameter, sporulation capacity, and percentage of germination demonstrated the high genetic variability of C. lindemuthianum. We also observed variation in conidial cytology. The conidia of most isolates showed septa formation after germination, in contrast to septa absence, previously reported in the literature. Sexual and asexual reproduction were evaluated for mechanisms that may contribute in the generation of variability in C. lindemuthianum. Conidial anastomosis tubes were commonly found, indicating that asexual reproduction can help increase variability in this species. Information from this study confirmed high variability in C. lindemuthianum and will guide future studies in basic knowledge and applied technologies.  相似文献   
100.
Moniliophthora perniciosa is a fungus that causes witches?? broom disease (WBD) in the cacao tree (Theobroma cacao). The M. perniciosa genome contains different transposable elements; this prompted an evaluation of the use of its retrotransposons as molecular markers for population studies. The inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) techniques were used to study the variability of 70?M. perniciosa isolates from different geographic origins and biotypes. A total of 43 loci was amplified. Cluster analysis of different geographical regions of C biotype revealed two large groups in the state of Bahia, Brazil. Techniques using retrotransposon-based molecular markers showed advantages over previously used molecular techniques for the study of genetic variability in M. perniciosa.  相似文献   
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