RESUMO
Swine dysentery, spirochetal colitis, and salmonellosis are production-limiting enteric diseases of global importance to the swine industry. Despite decades of efforts, mitigation of these diseases still relies on antibiotic therapy. A common knowledge gap among the 3 agents is the early B-cell response to infection in pigs. Thus, this study aimed to characterize the porcine B-cell response to Brachyspira hyodysenteriae, Brachyspira hampsonii (virulent and avirulent strains), Brachyspira pilosicoli, and Salmonella Typhimurium, the agents of the syndromes mentioned above. Immortalized porcine B-cell line derived from a crossbred pig with lymphoma were co-incubated for 8 h with each pathogen, as well as E. coli lipopolysaccharide (LPS) and a sham-inoculum (n = 3/treatment). B-cell viability following treatments was evaluated using trypan blue, and the expression levels of B-cell activation-related genes was profiled using reverse transcription quantitative PCR. Only S. Typhimurium and LPS led to increased B-cell mortality. B. pilosicoli downregulated B-lymphocyte antigen (CD19), spleen associated tyrosine Kinase (syk), tyrosine-protein kinase (lyn), and Tumour Necrosis Factor alpha (TNF-α), and elicited no change in immunoglobulin-associated beta (CD79b) and swine leukocyte antigen class II (SLA-DRA) expression levels, when compared to the sham-inoculated group. In contrast, all other treatments significantly upregulated CD79b and stimulated responses in other B-cell downstream genes. These findings suggest that B. pilosicoli does not elicit an immediate T-independent B-cell response, nor does it trigger antigen-presenting mechanisms. All other agents activated at least one trigger within the T-independent pathways, as well as peptide antigen presenting mechanisms. Future research is warranted to verify these findings in vivo.
Assuntos
Infecções por Bactérias Gram-Negativas , Doenças dos Suínos , Suínos , Animais , Infecções por Bactérias Gram-Negativas/patologia , Infecções por Bactérias Gram-Negativas/veterinária , Escherichia coli , Lipopolissacarídeos/toxicidade , Diarreia/veterinária , Diarreia/patologiaRESUMO
The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains under diagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acidSchiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, where as rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.(AU)
O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulinae inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células [...].(AU)
Assuntos
Animais , Galinhas/anatomia & histologia , Brachyspira/química , Brachyspira/imunologia , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/patologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterináriaRESUMO
The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains under diagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acidSchiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, where as rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.
O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulinae inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células [...].
Assuntos
Animais , Brachyspira/imunologia , Brachyspira/química , Galinhas/anatomia & histologia , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/patologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterináriaRESUMO
Introduction. In recent years, the Herbaspirillum genus has emerged as a pathogen in healthcare-related infections and has became stablished as an opportunistic pathogen.Hypothesis/Gap Statement. Little is known about the pathogenesis induced by Herbaspirillum genus.Aim. To evaluate the cytotoxic effects of genus Herbaspirillum, its ability to adhere to lung human cells and the ability of environmental and clinical strains of Herbaspirillum to induce pneumonia in mice.Methodology. Environmental and clinical isolates of Herbaspirillum were examined for their cytotoxic effects on the Calu-3 cell lineage. Cytotoxic activity of secretome was tested using MTT/neutral red assays and cell morphology analysis. Herbaspirillum adhesion on Calu-3 cells was assessed using bright-field microscopy and cell-associated bacteria were counted. A mouse model of acute lung infection was done using a clinical and an environmental strain. Adult male mice were used, and the pneumonia was inducted by intra-tracheal inoculation of 108 or 109 bacteria. Mice weight variations were evaluated at the end of the experiment. Bronchoalveolar lavage was collected and evaluated for total and differential cytology. A histological examination of lungs was performed giving a histological score.Results. The secretomes of all the strains induced morphological alterations in cells, but only H. seropedicae SmR1 were cytotoxic in MTT and neutral red assays. Clinical strains of H. frisingense AU14459 and H. hutttiense subsp. huttiense AU11883 exhibited low adherence to lung cells, while SmR1 was non-adhesive. Following intratracheal inoculation, mice treated with 109 c.f.u. of the SmR1 and AU11883 strains lost 18 and 6% of their weight over 7 days, respectively, and presented moderate clinical signs. Infected mice showed inflammatory cell infiltration in the perivascular and peribroncheal/peribronchiolar spaces. Bronchoalveolar fluid of mice inoculated with SmR1 109 c.f.u. presented an increase in total leucocyte cells and in neutrophils population.Conclusion. These in vivo and in vitro results provide insights into how some Herbaspirillum strains cause infection in humans, providing a basis for the characterization of pathogenesis studies on this emerging infectious agent.
Assuntos
Exossomos/metabolismo , Infecções por Bactérias Gram-Negativas/microbiologia , Herbaspirillum/patogenicidade , Pneumonia/microbiologia , Animais , Aderência Bacteriana , Líquido da Lavagem Broncoalveolar/citologia , Linhagem Celular , Sobrevivência Celular , Infecções por Bactérias Gram-Negativas/patologia , Herbaspirillum/isolamento & purificação , Herbaspirillum/metabolismo , Humanos , Pulmão/microbiologia , Pulmão/patologia , Masculino , Camundongos , Pneumonia/patologia , VirulênciaRESUMO
INTRODUCTION: Despite increasing application of molecular diagnostic methods for the detection of sexually transmitted infections, the cytological findings in pap smears of patients with pathogens that can be identified only by PCR are not yet well described. The aim of this study was to describe the most common cytological features in cervical pap smears of patients with Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum detected by multiplex PCR. METHODS: Cervical samples for conventional and liquid-based cytology and for multiplex PCR were collected from women ranging from 23 to 54 years old, who underwent routine screening at a gynecological Unit. RESULTS: Multiplex PCR was positive in 36.2% of the samples: Ureaplasma parvum 14.9%, Chlamydia trachomatis 10.6%, Trichomonas vaginalis 10.6%, Mycoplasma hominis 8.5%, Ureaplasma urealyticum 4.2%, Neisseria gonorrhoeae 2.1%, and Mycoplasma genitalium (0). Multiple pathogens were observed in 12.8% of samples. Microscopic cervicitis (≥10 polymorphonuclear leukocytes/epithelial cell) and normal (predominantly lactobacillary) microbiota were the most frequent findings in the samples in which the pathogens were detected alone or in multiple infections, except for samples with Trichomonas vaginalis in which the coccobacillary microbiota was the most common. In samples with microscopic cervicitis and normal microbiota, those with at least one pathogen identified by multiplex PCR were significantly more frequent than those with no pathogen, 66.6% versus 33.3%. CONCLUSION: Failure to identify an inflammatory agent in pap smear with intense neutrophil exudate may suggest the presence of Ureaplasma parvum, Ureaplasma urealyticum, Chlamydia trachomatis, or Trichomonas vaginalis. A remark on the intensity of inflammation should be made in the reports of cervical pap smears so that this cytological finding can be correlated with clinical and PCR results.
Assuntos
Colo do Útero/citologia , Colo do Útero/microbiologia , Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Ureaplasma urealyticum/genética , Adulto , Chlamydia trachomatis/genética , Feminino , Humanos , Programas de Rastreamento , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Mycoplasma/microbiologia , Mycoplasma hominis/genética , Neisseria gonorrhoeae/genética , Trichomonas vaginalis/genética , Ureaplasma/genética , Cervicite Uterina/microbiologia , Cervicite Uterina/patologia , Adulto JovemRESUMO
Aeromonas bacteria can cause an infection characterized by septicemia and is one of the most common pathogens in tropical fish. This disease is responsible for high morbidity and mortality rates, causing considerable losses in aquaculture. Thus, the understanding of its pathophysiology is crucial to develop control strategies of this bacterial infection in farmed fish. This study aimed to characterize early pathological aspects of acute sepsis in pacu (Piaractus mesopotamicus) experimentally infected with Aeromonas hydrophila. A total of 160 juvenile pacus were inoculated intraperitoneally with A. hydrophila (1.78 x 109 CFU/mL) and at 0 (control), 1, 3, 6, and 9 hours post-inoculation (hpi), animals were anesthetized and samples were collected for microbiological, light microscopy and transmission electron microscopy (TEM) analyzes. The results showed the occurrence of hemodynamic alterations, such as hemorrhage and congestion, which were observed mainly after 6 and 9 hpi. It was possible to re-isolate Aeromonas at all sampling times except in control group. However, just after 9 hpi it was possible to find the bacteria in all fish and tissues. Light microscopy analyses revealed a degenerative process, necrosis and vascular damage mainly at 6 and 9 hpi. According to the ultrastructural examination, areas of cellular death were identified in all examined tissues, especially at 6 and 9 hpi. However, the most severe, related to necrosis, were observed after 6 and 9 hpi. The findings suggested that this bacterium spreads in the first hpi through the fish organs, mainly affecting spleen, liver and kidney, causing irreversible lesions at the molecular level.
Assuntos
Aeromonas hydrophila , Caraciformes/microbiologia , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Sepse/veterinária , Animais , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Microscopia Eletrônica de Transmissão/veterinária , Sepse/microbiologia , Sepse/patologiaRESUMO
The prostate is the seat of three major causes of morbidity: benign prostatic hyperplasia, prostate cancer and prostatitis, three conditions in which inflammation has been implicated. A state of inflammation of the prostate gland, originally incited by an infection, an autoimmune response, a neurogenic stimulus or another trigger may have consequences on prostate functionality. In fact, male fertility depends intrinsically on the content of prostatic fluid factors secreted by the prostatic epithelium. Taking into account that the prostate gland is the major male accessory gland that exerts essential functions for male fertility, a state of local inflammation can alter male fertility by either directly impairing sperm quality or, indirectly, by causing prostate dysfunction. In the present review, we summarise the current knowledge regarding prostatitis due to well-known infections such as Escherichia coli, Chlamydia trachomatis and other commonly identified microorganisms focusing on inflammatory markers detected during these infections and seminal quality and male fertility alterations reported. We also focused on type III prostatitis or chronic nonbacterial prostatitis/chronic pelvic pain syndrome, of unknown aetiology, in which inflammation of an autoimmune origin, neurogenic stimuli or another trigger have been proposed and fertility alterations reported.
Assuntos
Fertilidade/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Infertilidade Masculina/imunologia , Próstata/imunologia , Prostatite/imunologia , Autoimunidade , Chlamydia trachomatis/imunologia , Chlamydia trachomatis/patogenicidade , Doença Crônica , Escherichia coli/imunologia , Escherichia coli/patogenicidade , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Humanos , Infertilidade Masculina/microbiologia , Infertilidade Masculina/patologia , Masculino , Próstata/microbiologia , Próstata/patologia , Prostatite/complicações , Prostatite/microbiologia , Prostatite/patologia , Sêmen/imunologia , Sêmen/microbiologiaRESUMO
Aeromonas dhakensis (Ad) CAIM 1873 growth was evaluated at different conditions and antibiotic susceptibility. Mortality and histopathological damages in hybrid tilapia Oreochromis niloticus × O. mossambicus, and virulence factors caused by Ad bacterial cells and extracellular products (ECPs) were evaluated, and the whole genome was obtained. Ad grew between 0.0 and 5.5% NaCl at a pH of between 4 and 10 and from 4 to 37°C. The lowest minimum inhibitory concentration was found for enrofloxacin (<5 µg ml-1), and bacteria were resistant to erythromycin, amoxicillin and ampicillin. Ad bacterial cells (1.86 × 105 cells g-1) and ECPs (0.462 µg protein fish-1) were highly virulent to challenged hybrid tilapia and caused over 80% mortality at 24 h. The primary clinical sign caused was haemorrhage, and damage was most marked in the spleen, liver, kidney and brain of fish challenged with bacterial cells. To our knowledge, this is the first report that Ad causes pyknotic and karyorrhectic nuclei of erythrocytes in the internal organs of hybrid tilapia, which was the most striking histopathological observation. The virulence of Ad to hybrid tilapia may be primarily related to the activity of haemolysins (hlyA genes) and cytotoxins (aerolysin aerA), along with the production of siderophores and proteases. We also found ß-lactamase, tetracycline and multiple antibiotic resistance genes, as well as adherence, iron acquisition, toxins (aerolysin family, haemolysins) and diverse protease genes.
Assuntos
Aeromonas/patogenicidade , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Tilápia/genética , Aeromonas/genética , Aeromonas/metabolismo , Animais , Doenças dos Peixes/patologia , Regulação Bacteriana da Expressão Gênica , Genoma Bacteriano , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , VirulênciaRESUMO
Digital dermatitis (DD) is one of the main causes of lameness in dairy cattle worldwide, and it is frequently reported in high-yielding, free stall dairy herds from regions with a temperate climate. However, DD is also observed with high prevalence in grazing cattle with a low milk yield in tropical regions. To clarify whether these differences have an impact on the etiology of the disease, we studied DD lesions from all year round grazing cattle of mixed breed in Brazil using high-throughput 16S rRNA gene sequencing and fluorescent in situ hybridization. The study included samples from 66 skin lesions and 5 healthy skins collected from five farms. Both techniques showed Treponema spp. to be the most abundant bacteria, present in all but one of the samples with minimal epidermal alterations. We identified eleven different Treponema strains belonging to the six major phylotypes of Treponema which have all previously been identified in DD lesions. Furthermore, we identify Dichelobacter nodosus in DD lesions by gene sequencing and also by fluorescent in situ hybridization in almost half of biopsy specimens in areas with mild epithelial damage and together with Treponema. The present data support the hypothesis that Treponema constitutes the main pathogen responsible for DD, independent of the environment and region where cows are kept, and it further suggests D. nodosus as another potentially important pathogen.
Assuntos
Criação de Animais Domésticos/métodos , Doenças dos Bovinos/microbiologia , Dichelobacter nodosus/patogenicidade , Dermatite Digital/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Infecções por Treponema/veterinária , Animais , Biópsia , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/patologia , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Dichelobacter nodosus/genética , Dichelobacter nodosus/isolamento & purificação , Dermatite Digital/epidemiologia , Dermatite Digital/patologia , Comportamento Alimentar , Feminino , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Herbivoria , Hibridização in Situ Fluorescente , Coxeadura Animal/epidemiologia , Coxeadura Animal/microbiologia , Coxeadura Animal/patologia , Ribotipagem , Treponema/genética , Treponema/isolamento & purificação , Infecções por Treponema/epidemiologia , Infecções por Treponema/microbiologia , Infecções por Treponema/patologiaRESUMO
Several pieces of evidence have linked the involvement of xanthine oxidase (XO), a source of uric acid and reactive oxygen species (ROS), to pro-oxidative and pro-inflammatory effects observed during bacterial fish diseases. Thus, the aim of this study was to evaluate whether upregulation of splenic XO activity contributes to disease pathogenesis of Aeromonas caviae infection, as well as whether it may be considered a pathway involved in ROS and nitric oxide (NO) production. XO activity increased in the spleen of infected animals, as did the splenic levels of uric acid, ROS and metabolites of nitric oxide (NOx), compared to the uninfected control group. Based on this evidence, upregulation of splenic XO activity induces pro-oxidant and pro-inflammatory profiles in the spleen of fish infected by A. caviae due to excessive formation of uric acid. Moreover, excessive uric acid induces the release of pro-inflammatory mediators, such as ROS and NOx, which contribute to disease pathophysiology. In summary, upregulation of XO activity may be considered a pathway involved in ROS and NOx production.
Assuntos
Aeromonas caviae/patogenicidade , Peixes-Gato/microbiologia , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/patologia , Óxido Nítrico/metabolismo , Ácido Úrico/metabolismo , Xantina Oxidase/metabolismo , Animais , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Inflamação/patologia , Oxirredução , Estresse Oxidativo , Baço/enzimologia , Baço/imunologia , Baço/patologiaRESUMO
Cytosolic and mitochondrial creatine kinases (CK), through the creatine kinase-phosphocreatine (CK/PCr) system, provide a temporal and spatial energy buffer to maintain cellular energy homeostasis. However, the effects of bacterial infections on the kidney remain poorly understood and are limited only to histopathological analyses. Thus, the aim of this study was to investigate the involvement of cytosolic and mitochondrial CK activities in renal energetic homeostasis in silver catfish experimentally infected with Aeromonas caviae. Cytosolic CK activity decreased in infected animals, while mitochondrial CK activity increased compared to uninfected animals. Moreover, the activity of the sodium-potassium pump (Na+, K+-ATPase) decreased in infected animals compared to uninfected animals. Based on this evidence, it can be concluded that the inhibition of cytosolic CK activity by A. caviae causes an impairment on renal energy homeostasis through the depletion of adenosine triphosphate (ATP) levels. This contributes to the inhibition of Na+, K+-ATPase activity, although the mitochondrial CK activity acted in an attempt to restore the cytosolic ATP levels through a feedback mechanism. In summary, A. caviae infection causes a severe energetic imbalance in infected silver catfish, which may contribute to disease pathogenesis.
Assuntos
Aeromonas caviae/patogenicidade , Peixes-Gato/microbiologia , Creatina Quinase Mitocondrial/metabolismo , Citosol/metabolismo , Metabolismo Energético/fisiologia , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Rim/metabolismo , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Brasil , Creatina Quinase/metabolismo , Citosol/enzimologia , Modelos Animais de Doenças , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Homeostase , Rim/microbiologia , Rim/patologia , Mitocôndrias/enzimologia , Mitocôndrias/metabolismo , Fosfocreatina/metabolismo , Fosforilação , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
BACKGROUND: Bovine respiratory disease (BRD) is an important problem in cattle production that is responsible for economic losses in dairy herds. Mycoplasma spp. are described as an important etiological agent of BRD. HYPOTHESIS: To evaluate the occurrence of the most important mycoplasmas in the lower respiratory tract of healthy and BRD cattle in relationship to clinical signs of BRD. ANIMALS: Sixty young dairy cattle were classified as healthy (n = 32) or cattle showing clinical signs of BRD (n = 28). METHODS: Tracheal lavage samples were collected and added to tubes containing Hayflick media. Mycoplasma spp. were identified by the presence of "fried egg" like colonies, biochemical tests and polymerase chain reaction (PCR). Occurrence of Mollicutes, M. bovis, M. mycoides subsp. mycoides SC and M. dispar was evaluated. The association between clinical signs of BRD and the presence of Mycoplasma spp. also was evaluated. RESULTS: Colonies were obtained from a 1-year-old BRD calf only. However, species identification was not possible. Mollicutes (P = .035) and M. dispar (P = .036) were more common in BRD cattle. The relationship between Mollicutes and crackle (P = .057) was not significant. M. dispar was associated to tachypnea (P = .045) and mixed dyspnea (P = .003). Relationships to heart rate (P = .062) and crackle (P = .062) were not significant. CONCLUSIONS AND CLINICAL IMPORTANCE: The results confirmed the importance of mycoplasma as an etiologic agent of BRD and suggested M. dispar as part of the respiratory microbiota and its possible role in the development of BRD.
Assuntos
Doenças dos Bovinos/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Infecções Respiratórias/veterinária , Tenericutes , Animais , Estudos de Casos e Controles , Bovinos , Doenças dos Bovinos/patologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Mycoplasma , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/patologia , Infecções por Mycoplasma/veterinária , Reação em Cadeia da Polimerase/veterinária , Infecções Respiratórias/microbiologia , Infecções Respiratórias/patologia , Tenericutes/patogenicidadeAssuntos
Aeromonas/isolamento & purificação , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Aquicultura , Argentina/epidemiologia , Proteínas de Bactérias/genética , Peixes-Gato , Surtos de Doenças/veterinária , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Proteínas Hemolisinas/genéticaAssuntos
Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Doenças dos Peixes/parasitologia , Animais , Aquicultura/métodos , Bactérias Gram-Negativas/isolamento & purificação , Infecções por Bactérias Gram-Negativas/patologia , Infecções por Bactérias Gram-Negativas/veterinária , Mesomycetozoea/isolamento & purificação , Infecções por Mesomycetozoea/patologia , Oncorhynchus mykiss/crescimento & desenvolvimento , PeruRESUMO
Despite the worldwide occurrence of Francisella noatunensis subsp. orientalis (Fno) infection in farmed tilapia, sensitivity and specificity estimates of commonly used diagnostic tests have not been reported. This study aimed to estimate the sensitivity and specificity of bacteriological culture and qPCR to detect Fno infection. We tested 559 fish, sampled from four farms with different epidemiological scenarios: (i) healthy fish in a hatchery free of Fno; (ii) targeted sampling of diseased fish with suggestive external clinical signs of francisellosis during an outbreak; (iii) convenience sampling of diseased and clinically healthy fish during an outbreak; and (iv) sampling of healthy fish in a cage farm without a history of outbreaks, but with francisellosis reported in other farms in the same reservoir. The qPCR had higher median sensitivity (range, 48.8-99.5%) than culture (range, 1.6-74.4%). Culture had a substantially lower median sensitivity (1.6%) than qPCR (48.8%) to detect Fno in carrier tilapia (farm 4). Median specificity estimates for both tests were >99.2%. The qPCR is the superior test for use in surveillance and monitoring programmes for francisellosis in farmed Nile tilapia, but both tests have high sensitivity and specificity which make them fit for use in the diagnosis of Fno outbreaks.
Assuntos
Ciclídeos , Doenças dos Peixes/microbiologia , Francisella/classificação , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Aquicultura , Brasil , DNA Bacteriano/genética , Surtos de Doenças/veterinária , Doenças dos Peixes/patologia , Francisella/genética , Francisella/isolamento & purificação , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e EspecificidadeRESUMO
UNLABELLED: Aeromonads are considered potential pathogens for humans and animals and are responsible for the etiology of intestinal and extraintestinal diseases. The presence of Aeromonas spp. in food and water shows that it is an important vehicle of infection in humans. The pathology caused by these bacteria involves several virulence factors, such as the ability to produce toxins, adhesion and invasion. The present study investigated the interaction of five Aeromonas caviae strains isolated from human diarrheic faeces with rabbit ileal and colonic mucosa ex vivo, using in vitro organ culture model. The in vitro adhesion assays using cultured tissue were performed with A. caviae strains co-incubated with intestinal fragments of ileum and colon over a period of 6 h. The fragments were analyzed by light and electron microscopy. All strains adhered to rabbit ileal and colonic mucosa ex vivo, with higher degree of adherence presented on colonic mucosa. The typical aggregative adherence pattern was observed among strains studied. Through electron and light microscopy, we observed extensive colonization of ileal and colonic mucosa, large mucus production, biofilm formation and morphological alterations such as intense vacuolization, structural disorganization, cell extrusion and destruction of the villi. These results demonstrate that in vitro organ culture of intestinal mucosa from rabbit may be used to investigate Aeromonas spp. PATHOGENESIS: Finally, our results support the pathogenic potential of Aeromonas emphasising their importance in public health.
Assuntos
Aeromonas caviae/citologia , Aderência Bacteriana/fisiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Mucosa Intestinal/microbiologia , Aeromonas caviae/genética , Aeromonas caviae/isolamento & purificação , Aeromonas caviae/patogenicidade , Animais , Biofilmes/crescimento & desenvolvimento , Modelos Animais de Doenças , Fezes/microbiologia , Humanos , Mucosa Intestinal/patologia , Coelhos , VirulênciaRESUMO
Aeromonas hydrophila is causing substantial economic losses in world aquaculture. This study determined the tolerance limit (LD50-96h) of A. hydrophila in Arapaima gigas, and also investigated the clinical signs after intradermal inoculation. Arapaima gigas fingerlings were inoculated intraperitoneally with 0 (control), 1.0×10(5), 1.0×10(6), 1.0×10(7), 1.0×10(9) and 1.0×10(10)CFU/mL of A. hydrophila for the determination of LD50-96h, which was 1.8×10(8)CFU/mL. In another trial with intradermal inoculation of 1.8×10(8)CFU/mL A. hydrophila, there was a 91.6% of mortality between 8 and 23h, and several clinical signs were found. As follows: depigmentation in the tegument, lesions in the tail and fins, loss of balance, reduction of respiratory movements, hemorrhagic foci, necrotic hemorrhages in the kidney, liver and swim bladder, splenomegaly, ascites in the abdominal cavity and hyperemia, enlargement of the gall bladder, among other clinical signs observed. The results showed that A. gigas has a relative tolerance to A. hydrophila when compared to other Neotropical fish species.
Assuntos
Aeromonas hydrophila/fisiologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Peixes , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Dose Letal Mediana , Rios , Clima TropicalRESUMO
Nephrotoxicity is the main adverse effect of colistin and polymyxin B (PMB). It is not clear whether these two antibiotics are associated with different nephrotoxicity rates. We compared the incidences of renal failure (RF) in patients treated with colistimethate sodium (CMS) or PMB for ≥48 h. A multicenter prospective cohort study was performed that included patients aged ≥18 years. The primary outcome was renal failure (RF) according to Risk, Injury, Failure, Loss, and End-stage renal disease (RIFLE) criteria. Multivariate analysis with a Cox regression model was performed. A total of 491 patients were included: 81 in the CMS group and 410 in the PMB group. The mean daily doses in milligrams per kilogram of body weight were 4.2 ± 1.3 and 2.4 ± 0.73 of colistin base activity and PMB, respectively. The overall incidence of RF was 16.9% (83 patients): 38.3% and 12.7% in the CMS and PMB groups, respectively (P< 0.001). In multivariate analysis, CMS therapy was an independent risk factor for RF (hazard ratio, 3.35; 95% confidence interval, 2.05 to 5.48;P< 0.001) along with intensive care unit admission, higher weight, older age, and bloodstream and intraabdominal infections. CMS was also independently associated with a higher risk of RF in various subgroup analyses. The incidence of RF was higher in the CMS group regardless of the patient baseline creatinine clearance. The development of RF during therapy was not associated with 30-day mortality in multivariate analysis. CMS was associated with significantly higher rates of RF than those of PMB. Further studies are required to confirm our findings in other patient populations.
Assuntos
Injúria Renal Aguda/induzido quimicamente , Antibacterianos/efeitos adversos , Colistina/análogos & derivados , Falência Renal Crônica/induzido quimicamente , Polimixina B/efeitos adversos , Injúria Renal Aguda/tratamento farmacológico , Injúria Renal Aguda/mortalidade , Injúria Renal Aguda/patologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/administração & dosagem , Peso Corporal , Colistina/administração & dosagem , Colistina/efeitos adversos , Esquema de Medicação , Feminino , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Negativas/patogenicidade , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/mortalidade , Infecções por Bactérias Gram-Negativas/patologia , Humanos , Unidades de Terapia Intensiva , Infecções Intra-Abdominais/tratamento farmacológico , Infecções Intra-Abdominais/microbiologia , Infecções Intra-Abdominais/mortalidade , Infecções Intra-Abdominais/patologia , Falência Renal Crônica/tratamento farmacológico , Falência Renal Crônica/mortalidade , Falência Renal Crônica/patologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Polimixina B/administração & dosagem , Estudos Prospectivos , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/microbiologia , Infecções Respiratórias/mortalidade , Infecções Respiratórias/patologia , Fatores de Risco , Análise de SobrevidaRESUMO
Injuries caused by freshwater stingray are common in several regions of South America, although they are underreported. The riverside inhabitants are the main victims in the Amazonian and Midwest regions of South America. The fishermen are injured mainly in the new focus of colonization of the rivers by freshwater stingrays. With the increasing population in these regions, where freshwater stingrays are found, there has been a significant increase in injuries within the general population. The highest increase occurred among tourists from other regions, where these animals are not known, when visiting these areas. The envenomations from the stingray causes prolonged and intense pain, both local and regionally. Generally these are associated with other local inflammatory manifestations, such as swelling and erythema. The injury often progresses to necrosis and it is considered potentially tetanogenic. A secondary infection is also a frequent local complication and most frequently is caused by Aeromonas species, usually Aeromonas hydrophila. Herein we report the first 2 cases of tetanus after freshwater stingray injuries: a 51-year-old men who had tetanus and recovered without sequel and the second a 67-year-old men who had severe tetanus and a deep, necrotizing soft-tissue infection with sepsis, septic shock and evolution to death.
Assuntos
Aeromonas , Mordeduras e Picadas/microbiologia , Coinfecção/patologia , Infecções por Bactérias Gram-Negativas/patologia , Rajidae , Tétano/patologia , Idoso , Animais , Brasil , Evolução Fatal , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Las infecciones por bacilos Gram negativos multiresistentes (BGN-MR) son frecuentes en nuestro hospital. Presentan limitadas opciones terapéuticas e importante impacto en la morbimortalidad y costos. Objetivo: analizar los factoes de riesgo y evolución de las bacteriemias por BGN-MR en pacientes neutropénicos febriles con patologías hematológicas. Materiales y métodos: estudio prospectivo, descriptivo y observacional de los factores de riesgo para BGN-MR en la población descripta. Se realizó análisis univariado y multivariado de variables clínicas, epidemiológicas, microbiológicas y evolutivas. Resultados: El 27 % de los episodios de neutropenia y fiebre cursaron con bacteriemias por BGN, 42 % de ellos fueron producidos por BGN-MR. En el análisis univariado, dichas bacteriemias se asociaron al uso previo de antibióticos; a las bacteriemias de brecha y neutropenias mayores a 7 días. En el análisis multivariado la bacteriemia de brecha mantuvo su significancia estadística (P<0,001; OR: 5,17; IC 95 % 2,1-12,7). Acinetobacter spp fue el BGN-MR más frecuentemente aislado incluso en los pacientes fallecidos. No se detectó el foco en el 45,9 % de los episodios. Los tratamientos inadecuados fueron significativamente más frecuentes en los pacientes con BGN-MR y la mortalidad tanto global como atribuible también se asoció significativamente al tratamiento inadecuado de las bacteriemias por BGN-MR (P<0,04;RR: 2,46;IC 95 % 1,03-5,9 y P< 0,014; RR: 3,02; IC 95 % 1,22-0,45 respectivamente). Conclusiones: Las bacteriemias por BGN-MR son frecuentes en la población estudiada en especial los que han recibido ATB previo y en las que surgen intratratamiento ATB. Recibieron con mayor frecuencia tratamiento empírico inadecuado, lo que se asoció a mayor mortalidad...
Bacterial infections by multiresistant Gram-negative bacilli (BGN-MR) are an increasing problem in our hospital with a major impact on morbidity, mortality and costs. Objective: to analize risk factors and outcome in bacteremia due to multiresistant Gram-negative bacilli in febrile neutropenic patients with hematologic diseases. Material and Methods: We conducted a prospective, descriptive and observational study to describe the risk factors and outcome of BGN-MR bacteremia in these patients. Results: Twenty seven percent of neutropenia and fever episodes had Gram-negative bacilli bacteremia and 42 % of them were caused by BGN-MR. Previous use of antibioteics, breakthrough bacteremia and prolonged neutropenia (<7 days) were significant in univariate analysis. In multivariate analysis only breakthrough bacteremia was significant (P< 0.001; OR 5,17;IC 95 % 2.1-12.7). Acinetobactersppp was the most common BGN-MR isolated in blood-stream infections and in patients who died. The source of infections was unknown in 45,9 % of the episodes. Inadequate empirical therapy was most common in BGN-MR bacteremia and it was associated with increased overall and attributable mortality (P<0.04; RR: 2.46; IC 95 % 1.03-5.9 y P<0.014; RR: 3.02; IC 95 % 1.22-7.45). Conclusions: BGN-MR was frequent in neutropenic patients with hematological diseases specially in those exposed to antibiotics and in breakthroug bacteremia. Inappropiate antimicrobial therapy was common and is associated with adverse outcome...