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ABSTRACT: Brucellosis is one of the most important zoonotic diseases in the world. Considering its strict zoonotic nature, understanding of the pathogenesis and immunity of Brucella spp. in natural animal hosts is essential to prevent human infections. Natural resistance against brucellosis has been demonstrated in cattle, and it is associated with the ability of macrophages to prevent intracellular replication of Brucella abortus. Identification of breeds that are resistant to B. abortus may contribute for controlling and eradicating brucellosis in cattle. This study aimed to compare macrophages from Nelore (Bos taurus indicus) or Holstein (Bos taurus taurus) regarding their resistance to B. abortus infection. Macrophages from Nelore were significantly more efficient in controlling intracellular growth of B. abortus when compared to Holstein macrophages even under intralysosomal iron restricting conditions. Furthermore, Nelore macrophages had higher transcription levels of inducible nitric oxide synthase (iNOS) and TNF-α at 12h post-infection (hpi) and higher levels of IL-12 at 24 hpi when compared to Holstein macrophages. Conversely, Holstein macrophages had higher levels of IL-10 transcripts at 24 hpi. Macrohages from Nelore also generated more nitric oxide (NO) in response to B. abortus infection when compared to Holstein macrophages. In conclusion, cultured Nelore macrophages are more effective in controlling intracellular replication of B. abortus, suggesting that Nelore cattle is likely to have a higher degree of natural resistance to brucellosis than Holstein.
Veterinary Immunology and Immunopathology 12/2012; · 2.08 Impact Factor
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ABSTRACT: The objective was to determine the effectiveness of various antimicrobial agents added to semen extender for inactivation of B. ovis or A. seminis in ovine semen after cryopreservation. In Experiment 1, 20 ejaculates from a crossbred ram infected with B. ovis were cryopreserved in Tris-based extenders with various antimicrobial agents: (I) control without antibiotics, (II) with penicillin and streptomycin (1000 IU/mL and 1 mg/mL, respectively), (III) lincomycin (0.15 mg/mL), (IV) sulphadiazine (0.60 mg/mL), and (V) gentamicin sulphate (0.25 mg/mL). Semen was stored in 0.25 mL straws at a final concentration of 150 × 10(6) spermatozoa/mL. After thawing (37 °C for 30 s), sperm total motility (TM), sperm morphology, integrity of sperm membranes, and bacterial growth were assessed. In Experiment 2, six B. ovis isolates were separately inoculated into aliquots of a fresh ejaculate from a B. ovis-free ram. Mock inoculated semen was processed for cryopreservation using the five extenders described above, and bacteriologically evaluated after thawing. In Experiment 3, sensitivity of A. seminis to the same antimicrobial agents was evaluated by inoculating an ejaculate from an A. seminis and B. ovis-free ram. There were no significant differences among treatments in post-thawing sperm parameters. B. ovis was isolated from 100% (20/20), 0% (0/20), 95% (19/20), 100% (20/20), and 5% (1/20) of semen samples diluted in tris-based extender of untreated (I) and treated semen samples with antimicrobial agents II, III, IV, and V, respectively. Frequencies of isolation from samples treated with antimicrobial agent II and V were significantly lower than untreated ones (P < 0.05). There were no significant differences in the profile of antimicrobial resistance of different B. ovis isolates. A. seminis had a similar sensitivity to the antimicrobial agents. We concluded that addition of a combination of penicillin and streptomycin or gentamicin alone to ram semen cryo-extenders inactivated B. ovis and A. seminis.
Theriogenology 11/2010; 74(8):1476-81. · 1.96 Impact Factor
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ABSTRACT: The goal of this study was to morphologically characterize a ligated ileal loop model of Salmonella enterica serotype Typhimurium infection in rhesus macaques (Macaca mulatta) and to verify the occurrence of Salmonella-induced cell death in vivo. Eight adult healthy male rhesus macaques were used for ligated ileal loop surgery. Four macaques had been intravenously inoculated with simian immunodeficiency virus (SIV) mac251. Ileal ligated loops were inoculated with wild-type (WT) S. Typhimurium strain IR715 (ATCC14028 nal (r)), an isogenic noninvasive mutant strain (ATCC14028 nal (r) ΔsipAΔsopABDE2), or sterile Luria Bertani broth. Loops were surgically removed at 2, 5, and 8 hours post-inoculation (hpi). Intestinal samples were processed for histopathology, immunohistochemistry for detecting Salmonella, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and transmission electron microscopy. Combined histopathology scores were similar between SIV-infected and control macaques. As expected, the invasion-deficient mutant was less pathogenic than WT S. Typhimurium. Neutrophil infiltrate in the intestinal mucosa correlated with bacterial loads (r = 0.7148; P < .0001) and fluid accumulation (r = 0.6019; P < .0001) in the lumen of the intestinal loops. Immunolabeled WT S. Typhimurium was observed in the epithelium and lamina propria at the tip of the villi at 2 hpi, progressing toward deeper lamina propria at 5-8 hpi. Most TUNEL-positive cells localized to the lamina propria, and some had morphological features of macrophages. Ultrastructurally, bacteria were observed intracellularly in the lamina propria as well as within apoptotic bodies. This study provides morphological evidence of Salmonella-induced cell death in vivo in a relevant nonhuman primate model.
Veterinary Pathology 11/2010; 48(5):933-41. · 1.95 Impact Factor
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Equine Veterinary Education. 01/2010; 12(5):256 - 259.
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ABSTRACT: This report describes a pathological, immunohistochemical and bacteriological study of 42 cows and their progeny (aborted fetuses, weak premature calves, and healthy full-term calves) infected at 6-7 months of gestation by conjunctival inoculation with Brucella abortus. Samples were collected at necropsy within 48 h of abortion or parturition. The most significant lesions were necrotizing and suppurative placentitis and lymphohistiocytic mastitis in cows, and fibrinous pleuritis, fibrinous pericarditis and bronchopneumonia in aborted fetuses. B. abortus was isolated more frequently from milk samples than from mammary tissues, and milk samples from cows with mastitis were often infected. Organisms were often demonstrated immunohistochemically and by culture in tissues showing moderate to severe histological changes.
Journal of comparative pathology 01/2009; 140(2-3):149-57. · 1.73 Impact Factor
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ABSTRACT: We report experimental results concerning observation of a pattern forming system, subject to directional viscous fingering (printer's instability). This system was excited by a time-dependent, periodic perturbation. A variety of spatiotemporal effects was observed, including pattern transient dynamics, wave vector selection, and morphological transitions. Detailed measurement of pattern shape and its associated Fourier modes assured the detection of a crossover between different regimes of the pattern evolution.
Physical Review E 06/2008; 77(6 Pt 2):066310. · 2.26 Impact Factor
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ABSTRACT: A precursor is a kind of instability in a nonequilibrium system observed prior to a bifurcation. It has been observed in important natural phenomena such as earthquakes and epileptic seizures. In most cases just qualitative results were reported, with the exception of some quantitative observations of noisy precursors. We report the observation of precursors excited by a deterministic perturbation. Quantitative characterization of this type of precursor is important in predicting a bifurcation in advance. This predicting ability comes from a dynamical structure present on the perturbed system.
Physical Review E 11/2006; 74(4 Pt 1):041104. · 2.26 Impact Factor
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ABSTRACT: A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis.
Veterinary Parasitology 10/2006; 140(3-4):356-61. · 2.58 Impact Factor
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ABSTRACT: Although visceral leishmaniasis is primarily transmitted by a biological invertebrate vector, transmission in the absence of the vector has been reported, including venereal transmission in humans. Considering the possibility of venereal transmission, we studied genital lesions in dogs naturally infected with visceral leishmaniasis and shedding of Leishmania sp. in the semen. Approximately 200 dogs were serologically tested for anti-Leishmania antibodies and divided into three groups: 1) serologically negative dogs (n = 20), 2) asymptomatic serologically positive dogs (n = 20), and 3) symptomatic serologically positive dogs (n = 20). Samples from both testes, all segments of both epididymes, prostate gland, glans penis, and prepuce were histologically evaluated and processed for immunodetection of Leishmania sp. Semen samples were obtained from 22 symptomatic serologically positive dogs and processed for detecting Leishmania DNA by polymerase chain reaction. A significantly higher frequency of inflammation was observed in the epididymes, glans penis, and prepuce of dogs with visceral leishmaniasis, which was associated with a high frequency of immunohistochemically positive tissues (up to 95% of tissues from symptomatic dogs were positive by immunohistochemistry). Leishmania DNA was detected in eight of 22 semen samples from symptomatic dogs. Together these findings indicate that genital lesions and shedding of Leishmania sp. (donovani complex) in the semen are associated with visceral leishmaniasis. Additional studies should address the possibility of venereal transmission of the disease in the dog.
Veterinary Pathology 10/2005; 42(5):650-8. · 1.95 Impact Factor
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ABSTRACT: The yield as well as phenotypic and functional parameters of canine peripheral blood monocyte-derived macrophages were analyzed. The cells that remained adherent to Teflon after 10 days of culture had high phagocytic activity when inoculated with Leishmania chagasi. Flow cytometric analysis demonstrated that more than 80% of cultured cells were positive for the monocyte/macrophage marker CD14.
Memórias do Instituto Oswaldo Cruz 09/2005; 100(5):521-4. · 2.15 Impact Factor
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ABSTRACT: Morphological changes in the placenta at periparturition and the involvement of apoptosis controlling cell populations in the placentome were evaluated in Holstein and Nelore cows. Twenty-two Holstein cows at the end of gestation with normal non-induced parturition (group I) and 10 Holstein cows with induced parturition (group II) were used in this study. In addition, groups III and IV were constituted by 10 Nelore cows at the end of gestation with normal non-induced parturition and 21 Nelore cows with induced parturition, respectivelly. Binucleate trophoblastic cells, epithelial caruncular cells and apoptotic bodies were quantified by light microscopy. Apoptosis was further evaluated by transmission electron microscopy, ELISA and DNA electrophoresis in agarose gel. The number of caruncular epithelial cells did not vary between breeds with normal or induced parturition. The number of binucleate cells was significantly different between Holstein and Nelore cows with normal non-induced parturition (groups I vs. III), but their numbers were not significantly different between breeds when parturition was induced. In cows with normal non-induced parturition, the intensity of apoptosis, as assessed by morphometric analysis and ELISA, was significantly higher in group I when compared to group III. However, no difference in the intensity of apoptosis was found between groups II and IV. Characteristic apoptotic ladder pattern of DNA fragmentation was detected by agarose gel electrophoresis. Ultra-structural features of apoptosis in maternal and fetal epithelium as well as capillary vascular endothelium were identified by transmission electron microscopy. In conclusion, in Nelore placental maturation occurs precociously when compared to Holstein. In both breeds, apoptosis is an active physiological event that seems to be required for maturation and normal post-partum release of the placenta.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2004;
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ABSTRACT: Infections with Salmonella serotypes are a major cause of food-borne diseases worldwide. Animal models other than the mouse have been employed for the study of nontyphoidal Salmonella infections because the murine model is not suitable for the study of Salmonella-induced diarrhea. The microbe has developed mechanisms to exploit the host cell machinery to its own purpose. Bacterial proteins delivered directly into the host cell cytosol cause cytoskeletal changes and interfere with host cell signaling pathways, which ultimately enhance disease manifestation. Recently, marked advances have been made in our understanding of the molecular interactions between Salmonella serotypes and their hosts. Here, we discuss the molecular basis of the pathogenesis of Salmonella-induced enteritis.
Brazilian Journal of Medical and Biological Research 02/2003; 36(1):3-12. · 1.13 Impact Factor
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ABSTRACT: Os efeitos do succinato sódico de hidrocortisona (SSH) nas lesões de isquemia e reperfusão no jejuno foram estudados em 12 eqüinos submetidos a isquemia total arteriovenosa e venosa no jejuno. Após uma hora de isquemia, seis eqüinos receberam 4,0mg/kg/IV de SSH (grupo T) e os demais receberam placebo (grupo NT). Foram colhidas amostras para avaliação histomorfológica após uma e duas horas de isquemia e uma, duas e 12 horas de reperfusão, sendo as alterações quantificadas por meio de escores. Os escores para infiltração de neutrófilos, edema e hemorragia foram equivalentes entre os grupos T e NT. No segmento submetido a isquemia venosa o agravamento da lesão na mucosa durante a reperfusão foi significativo (P<0,05) apenas no grupo NT, indicando que o SSH atenuou a lesão de reperfusão no jejuno. Após 12 horas de reperfusão os escores para lesão na mucosa na isquemia arteriovenosa foram equivalentes entre os grupos T e NT. Na isquemia venosa os escores foram significativamente menores no grupo T (P<0,01), indicando que o tratamento com SSH acelerou a reparação das lesões. Estes resultados indicam que o SSH pode ser utilizado como tratamento adjuvante de afecções isquêmicas e de reperfusão no eqüino.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2003;
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ABSTRACT: Foram estudadas características da bioquímica do sangue, da pressão arterial e da freqüência de pulso de 12 bezerros mantidos sob anestesia por 13 horas, utilizando-se propofol para a indução e isofluorano para manutenção, associados à administração de morfina intra-tecal. Os valores de freqüência de pulso, pressão arterial e glicemia apresentaram pequenas variações e se mantiveram próximos dos valores de referência para bezerros anestesiados. Ao longo do período de anestesia houve aumento significativo, mas discreto, do hematócrito, hemoglobina, pCO2, CO2 total, bicarbonato e potássio. O pH do sangue, pO2, Na+ e Ca++ apresentaram reduções significativas. Este protocolo anestésico foi seguro para a manutenção de bezerros anestesiados por período prolongado.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2003;
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ABSTRACT: The host response to Salmonella plays a major role in the outcome of infection. The present study was undertaken to further characterize Salmonella typhimurium infection in neonatal calves at both the morphologic and the molecular level using the ligated ileal loop model. Eight 4-5-week-old male Holstein calves underwent laparotomy, and loops were prepared in the ileum. The loops were either inoculated with an S. typhimurium strain pathogenic for cattle or injected with sterile LB broth as control. Samples for histology, transmission and scanning electron microscopy, and RNA extraction were collected at various time points between 5 minutes and 12 hours postinfection. Invasion of both M cells and enterocytes began at 15 minutes postinfection. No specific cell type was the main target for invasion. Intracellular bacteria were observed in the lamina propria after 1 hour postinfection. A severe acute neutrophilic response was associated with invasion of the Peyer's patches. Upregulated expression of CXC chemokines (interleukin [IL]-8, growth-related oncogenes, [GRO] alpha and gamma, and granulocyte chemotactic protein [GCP]2) was detected by reverse transcription polymerase chain reaction beginning at 1 hour postinfection. Expression of proinflammatory (IL-1beta, IL-18, and tumor necrosis factor [TNF]alpha) and anti-inflammatory (IL-10, IL-IRa, and IL-4) cytokines was also assessed. A marked increase in expression of IL-1beta was observed, whereas the profile of expression of IL-18 and TNFalpha did not change after infection. Upregulation of IL-1Ra and IL-4 but not of IL-10 was observed. These findings indicate that infection of bovine ligated ileal loops with S. typhimurium results in an acute neutrophilic inflammatory response that is associated with the upregulation of CXC chemokines (IL-8, GROalpha and gamma, and GCP2), IL-1beta, IL-IRa, and IL-4.
Veterinary Pathology 04/2002; 39(2):200-15. · 1.95 Impact Factor
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ABSTRACT: Salmonella enterica serovar Typhimurium causes cell death in bovine monocyte-derived and murine macrophages in vitro by a sipB-dependent mechanism. During this process, SipB binds and activates caspase-1, which in turn activates the proinflammatory cytokine interleukin-1beta through cleavage. We used bovine ileal ligated loops to address the role of serovar Typhimurium-induced cell death in induction of fluid accumulation and inflammation in this diarrhea model. Twelve perinatal calves had 6- to 9-cm loops prepared in the terminal ileum. They were divided into three groups: one group received an intralumen injection of Luria-Bertani broth as a control in 12 loops. The other two groups (four calves each) were inoculated with 0.75 x 10(9) CFU of either wild-type serovar Typhimurium (strain IR715) or a sopB mutant per loop in 12 loops. Hematoxylin and eosin-stained sections were scored for inflammation, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells were detected in situ. Fluid accumulation began at 3 h postinfection (PI). Inflammation was detected in all infected loops at 1 h PI. The area of TUNEL-labeled cells in the wild-type infected loops was significantly higher than that of the controls at 12 h PI, when a severe inflammatory response and tissue damage had already developed. The sopB mutant induced the same amount of TUNEL-positive cells as the wild type, but it was attenuated for induction of fluid secretion and inflammation. Our results indicate that serovar Typhimurium-induced cell death is not required to trigger an early inflammatory response and fluid accumulation in the ileum.
Infection and Immunity 08/2001; 69(7):4610-7. · 4.16 Impact Factor
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ABSTRACT: It was previously demonstrated that Salmonella enterica serovar Typhimurium induces cell death with features of apoptosis in murine macrophages. Mice infected with Salmonella serovar Typhimurium develop systemic disease without diarrhea, whereas the infection in cattle and in humans is localized and characterized by diarrhea. Considering these clinical disease expression differences between mice and cattle, we investigated whether serovar Typhimurium is cytotoxic for bovine macrophages. Macrophages infected with serovar Typhimurium grown in the logarithmic phase quickly underwent cell death. Macrophages infected with stationary-phase cultures or with a mutant lacking sipB underwent no immediate cell death but did develop delayed cytotoxicity, undergoing cell death between 12 and 18 h postinfection. Both pathways were temporarily blocked by the general caspase inhibitor Z-VAD-Fmk and by the caspase 1 inhibitor Z-YVAD-Fmk. Comparisons of macrophages from cattle naturally resistant or susceptible to intracellular pathogens indicated no differences between these two genetic backgrounds in terms of susceptibility to serovar Typhimurium-induced cell death. We conclude that Salmonella serovar Typhimurium induces cell death in bovine macrophages by two distinct mechanisms, early sipB-mediated and delayed sipB-independent mechanisms.
Infection and Immunity 05/2001; 69(4):2293-301. · 4.16 Impact Factor
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ABSTRACT: The objective of this report was to describe the pathological changes in the urinary bladder of female rats naturally infected with Trichosomoides crassicauda. Forty eight 5 to 8 months of age female Wistar rats were studied. At necropsy uroliths were detected in seven animals (14.6%). Among the bladders that contained the parasite only three (6.3%) did not show any significant histological change and were considered normal, but on the contrary, three (6.3%) did contain papillomas and uroliths, four (8.3%) had uroliths associated with epithelial hyperplasia ranging from moderate to severe, and the remaining (79.2%) showed variable degrees of epithelial hyperplasia and inflammation without urolithiasis. It cannot be concluded whether irritation of the epithelium by the parasite itself plays a role in the formation of papillomas. However, the absence of papillomas in animals without uroliths suggests a positive relationship between uroliths and papillomas.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2001;
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ABSTRACT: Sob anestesia geral, com constante controle sobre a pressão arterial e a saturação de oxigênio da hemoglobina arterial, realizou-se celiotomia em 12 eqüinos. No cólon menor exposto foram demarcados três segmentos de 25cm, separados entre si por igual distância. Dois desses segmentos foram submetidos à isquemia arteriovenosa completa por 90 (grupo A) ou 180 minutos (grupo B). O terceiro segmento foi o grupo-controle. Amostras para histopatologia foram colhidas ao final dos períodos de isquemia e após 90 e 180 minutos de reperfusão no grupo A e após 90 minutos de reperfusão no grupo B. No controle, colheram-se amostras no início e final do procedimento. Avaliaram-se as lesões produzidas na mucosa e na submucosa pelos métodos semiquantitativos-escores para desprendimento de epitélio, edema, hemorragia e infiltrado de neutrófilos, e pelos quantitativos-porcentagem de perda de mucosa (PM) e razão cripta:interstício (C:I). As lesões isquêmicas foram mais intensas no grupo B do que no A para PM, C:I, desprendimento de epitélio e edema de mucosa. As amostras obtidas após a reperfusão revelaram que houve agravamento na PM, C:I, desprendimento de epitélio e edema de submucosa em ambos os grupos. Concluiu-se que a reperfusão agravou as lesões isquêmicas no cólon menor e que o modelo proposto é viável para produção dessas lesões.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2001;
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ABSTRACT: Estudaram-se a ocorrência e a predisposição etária de neoplasias testiculares em cães. Em 497 cães necropsiados, 47 apresentaram tumores testiculares, correspondendo à frequência de 9,45% (47/497). Houve aumento significativo na freqüência de tumores testiculares em animais velhos.
Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 01/2000;
