RESUMEN
Under homoeostatic conditions, the relationship between the coral Pocillopora damicornis and Vibrio coralliilyticus is commensal. An increase in temperature, or in the abundance of V. coralliilyticus, can turn this association pathogenic, causing tissue lysis, expulsion of the corals' symbiotic algae (genus Symbiodinium), and eventually coral death. Using a combination of microfluidics, fluorescence microscopy, stable isotopes, electron microscopy and NanoSIMS isotopic imaging, we provide insights into the onset and progression of V. coralliilyticus infection in the daytime and at night, at the tissue and (sub-)cellular level. The objective of our study was to connect the macro-scale behavioural response of the coral to the micro-scale nutritional interactions that occur between the host and its symbiont. In the daytime, polyps enhanced their mucus production, and actively spewed pathogens. Vibrio infection primarily resulted in the formation of tissue lesions in the coenosarc. NanoSIMS analysis revealed infection reduced 13C-assimilation in Symbiodinium, but increased 13C-assimilation in the host. In the night incubations, no mucus spewing was observed, and a mucus film was formed on the coral surface. Vibrio inoculation and infection at night showed reduced 13C-turnover in Symbiodinium, but did not impact host 13C-turnover. Our results show that both the nutritional interactions that occur between the two symbiotic partners and the behavioural response of the host organism play key roles in determining the progression and severity of host-pathogen interactions. More generally, our approach provides a new means of studying interactions (ranging from behavioural to metabolic scales) between partners involved in complex holobiont systems, under both homoeostatic and pathogenic conditions.
Asunto(s)
Antozoos/microbiología , Simbiosis , Vibrio/fisiología , Animales , Antozoos/anatomía & histología , Antozoos/metabolismo , Antozoos/fisiología , Conducta Animal , Dinoflagelados/metabolismo , Interacciones Huésped-Patógeno , Nutrientes , TemperaturaRESUMEN
BACKGROUND: Global warming has triggered an increase in the prevalence and severity of coral disease, yet little is known about coral/pathogen interactions in the early stages of infection. The point of entry of the pathogen and the route that they take once inside the polyp is currently unknown, as is the coral's capacity to respond to infection. To address these questions, we developed a novel method that combines stable isotope labelling and microfluidics with transmission electron microscopy (TEM) and nanoscale secondary ion mass spectrometry (NanoSIMS), to monitor the infection process between Pocillopora damicornis and Vibrio coralliilyticus under elevated temperature. RESULTS: Three coral fragments were inoculated with 15N-labeled V. coralliilyticus and then fixed at 2.5, 6 and 22 h post-inoculation (hpi) according to the virulence of the infection. Correlative TEM/NanoSIMS imaging was subsequently used to visualize the penetration and dispersal of V. coralliilyticus and their degradation or secretion products. Most of the V. coralliilyticus cells we observed were located in the oral epidermis of the fragment that experienced the most virulent infection (2.5 hpi). In some cases, these bacteria were enclosed within electron dense host-derived intracellular vesicles. 15N-enriched pathogen-derived breakdown products were visible in all tissue layers of the coral polyp (oral epidermis, oral gastrodermis, aboral gastrodermis), at all time points, although the relative 15N-enrichment depended on the time at which the corals were fixed. Tissues in the mesentery filaments had the highest density of 15N-enriched hotspots, suggesting these tissues act as a "collection and digestion" site for pathogenic bacteria. Closer examination of the sub-cellular structures associated with these 15N-hotspots revealed these to be host phagosomal and secretory cells/vesicles. CONCLUSIONS: This study provides a novel method for tracking bacterial infection dynamics at the levels of the tissue and single cell and takes the first steps towards understanding the complexities of infection at the microscale, which is a crucial step towards understanding how corals will fare under global warming.
Asunto(s)
Enfermedades de los Animales/microbiología , Antozoos/microbiología , Microfluídica/métodos , Espectrometría de Masa de Ion Secundario/métodos , Espectrometría de Masa de Ion Secundario/veterinaria , Vibriosis/microbiología , Vibriosis/veterinaria , Vibrio/patogenicidad , Animales , Antozoos/citología , Antozoos/inmunología , Células Epidérmicas/microbiología , Células Epidérmicas/patología , Epidermis/microbiología , Epidermis/patología , Calentamiento Global , Marcaje Isotópico , Israel , Microscopía Electrónica de Transmisión , Temperatura , Vibriosis/patología , VirulenciaRESUMEN
PURPOSE: We report the largest series of renal embolizations performed for a variety of indications. PATIENTS AND METHODS: A retrospective analysis was performed on embolizations performed in our institution from 1997 to 2002 encompassing 36 patients who underwent 44 procedures. RESULTS: Embolization was successful on the first attempt in 87% of the patients. A second embolization was performed in four of the five unsuccessful cases, three successfully, increasing the success rate to 95%. The mean postoperative narcotic use was 27.2 mg of morphine equivalent, and 10 mg or less was required by 45% of the patients. In the 14 patients who had not also undergone a surgical procedure, the mean narcotic use was 21 mg, and 64% required 10 mg or less. Only 15% of the patients developed fever, which resolved within 2 days in all cases. Leukocytosis was seen in 47%. Follow-up creatinine and hypertension information was available in 16 and 18 patients, respectively. After a mean follow-up of 269 days, only one patient had a clinically significant rise in the creatinine concentration. After a mean follow-up of 496 days, two patients had new-onset hypertension. There was no statistically significant difference in the success rate, narcotic use, complications, creatinine concentrations, or the likelihood of fever, leukocytosis, or hypertension according to the indication for embolization or the agent used. Use of a microcatheter was associated with less parenchymal loss, and decreased parenchymal loss was associated with a significant reduction of narcotic use. CONCLUSIONS: Renal embolization is a highly effective and well-tolerated procedure in a variety of urologic conditions. The indications and material used did not have a significant effect on the outcome. Reducing parenchymal loss can significantly reduce morbidity.
Asunto(s)
Embolización Terapéutica/efectos adversos , Embolización Terapéutica/estadística & datos numéricos , Enfermedades Urológicas/terapia , Analgésicos/uso terapéutico , Creatinina/sangre , Fiebre/etiología , Humanos , Hipertensión/etiología , Leucocitosis/etiología , Morbilidad , Evaluación de Procesos y Resultados en Atención de Salud , Radiografía , Reoperación/estadística & datos numéricos , Estudios Retrospectivos , Tiempo , Resultado del Tratamiento , Enfermedades Urológicas/complicaciones , Enfermedades Urológicas/diagnóstico por imagenRESUMEN
UNLABELLED: Hemorrhagic hypotension may aggravate the detrimental effects of head trauma on neurologic outcome. Our study examined whether using phenylephrine or large volumes of saline IV to increase mean arterial blood pressure (MAP) to 70, 80, or 90 mm Hg during the combination of head trauma and uncontrolled hemorrhage would improve neurologic outcome. Rats were assigned to one of 17 groups. In Groups 1-5, the variables were head trauma (yes/no), hemorrhage (yes/no), 0 or 3 mL saline per milliliter of blood lost, and no target MAP. In Groups 6-11, hemorrhage was or was not combined with head trauma, and large volumes of saline were given IV to achieve target MAPs of 70, 80, or 90 mm Hg. Groups 12-17 were similar to Groups 6-11 except that phenylephrine was used rather than saline to achieve target MAPs. Saline increased blood loss at 2 h to approximately 16 and 25 mL at a MAP of 80 and 90 mm Hg respectively, increased (worsened) the neurodeficit score but not cerebral edema at 24 h, and decreased survival rate at 2 and 24 h. Because phenylephrine was fatal for 62 of 63 rats, group mean values for blood loss, neurodeficit score, and brain tissue specific gravity could not be calculated. We conclude that supporting MAP with either phenylephrine or large volumes of saline worsened the neurodeficit score and/or survival and did not affect cerebral edema formation in our rat model of head trauma combined with hemorrhage. IMPLICATIONS: The results of this study indicate that maintaining mean arterial blood pressure at 70, 80, or 90 mm Hg with either phenylephrine or large volumes of saline worsened the neurodeficit score and/or survival and did not affect cerebral edema formation in our rat model of head trauma combined with hemorrhage.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Encefalopatías/etiología , Traumatismos Cerrados de la Cabeza/complicaciones , Hemorragia/complicaciones , Hipotensión/tratamiento farmacológico , Análisis de Varianza , Animales , Encéfalo/fisiopatología , Edema Encefálico/etiología , Edema Encefálico/fisiopatología , Modelos Animales de Enfermedad , Fluidoterapia , Traumatismos Cerrados de la Cabeza/fisiopatología , Hemorragia/fisiopatología , Hipotensión/fisiopatología , Infusiones Intravenosas , Masculino , Fenilefrina/uso terapéutico , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Gravedad Específica , Tasa de Supervivencia , Factores de Tiempo , Vasoconstrictores/uso terapéutico , Heridas no Penetrantes/complicaciones , Heridas no Penetrantes/fisiopatologíaRESUMEN
Formalin-killed Streptococcus difficile strains used as vaccines delivered intraperitoneally were able to protect tilapia against a challenge of 100 LD50. The protection obtained was not strain specific. A vaccine based on an S. difficile extract containing 50% protein conjugated to alum also protected tilapia challenged with a virulent S. difficile strain. Protection in tilapia was correlated with the development of specific agglutinins. Western blot analysis supported the hypothesis that only a few proteins act as protective antigens in both the whole-cell vaccine and the streptococcal extract. The high efficacy of these vaccines make them good candidates for the control of streptococcal fish meningoencephalitis.