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1.
Gastroenterology ; 153(6): 1594-1606.e2, 2017 12.
Artículo en Inglés | MEDLINE | ID: mdl-28865734

RESUMEN

BACKGROUND & AIMS: Separation of newborn rats from their mothers induces visceral hypersensitivity and impaired epithelial secretory cell lineages when they are adults. Little is known about the mechanisms by which maternal separation causes visceral hypersensitivity or its relationship with defects in epithelial secretory cell lineages. METHODS: We performed studies with C3H/HeN mice separated from their mothers as newborns and mice genetically engineered (Sox9flox/flox-vil-cre on C57BL/6 background) to have deficiencies in Paneth cells. Paneth cell deficiency was assessed by lysozyme staining of ileum tissues and lysozyme activity in fecal samples. When mice were 50 days old, their abdominal response to colorectal distension was assessed by electromyography. Fecal samples were collected and microbiota were analyzed using Gut Low-Density Array quantitative polymerase chain reaction. RESULTS: Mice with maternal separation developed visceral hypersensitivity and defects in Paneth cells, as reported from rats, compared with mice without maternal separation. Sox9flox/flox-vil-Cre mice also had increased visceral hypersensitivity compared with control littermate Sox9flox/flox mice. Fecal samples from mice with maternal separation and from Sox9flox/flox-vil-cre mice had evidence for intestinal dysbiosis of the microbiota, characterized by expansion of Escherichia coli. Daily gavage of conventional C3H/HeN adult mice with 109 commensal E coli induced visceral hypersensitivity. Conversely, daily oral administration of lysozyme prevented expansion of E coli during maternal separation and visceral hypersensitivity. CONCLUSIONS: Mice with defects in Paneth cells (induced by maternal separation or genetically engineered) have intestinal expansion of E coli leading to visceral hypersensitivity. These findings provide evidence that Paneth cell function and intestinal dysbiosis are involved in visceral sensitivity.


Asunto(s)
Ansiedad de Separación/complicaciones , Escherichia coli/crecimiento & desarrollo , Microbioma Gastrointestinal , Hiperalgesia/etiología , Células de Paneth/microbiología , Dolor Visceral/etiología , Factores de Edad , Animales , Animales Recién Nacidos , Ansiedad de Separación/metabolismo , Ansiedad de Separación/microbiología , Ansiedad de Separación/fisiopatología , Modelos Animales de Enfermedad , Disbiosis , Heces/microbiología , Femenino , Predisposición Genética a la Enfermedad , Hiperalgesia/metabolismo , Hiperalgesia/microbiología , Hiperalgesia/fisiopatología , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Noqueados , Muramidasa/administración & dosificación , Muramidasa/metabolismo , Células de Paneth/metabolismo , Fenotipo , Factor de Transcripción SOX9/genética , Factor de Transcripción SOX9/metabolismo , Dolor Visceral/metabolismo , Dolor Visceral/microbiología , Dolor Visceral/fisiopatología
2.
Parasitol Res ; 114(10): 3815-26, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26143866

RESUMEN

Cattle besnoitiosis due to Besnoitia besnoiti is spreading across Europe and is responsible for severe economic losses in newly infected herds. Experimentally speaking, rabbits have been found to be susceptible to this parasite. The adaptation of B. besnoiti to rabbits may offer a new, easier and cheaper model of investigation for this disease. This study compared the virulence between tachyzoites and bradyzoites of B. besnoiti in rabbits. Eighteen New Zealand rabbits were allocated into three groups of six animals each. The rabbits from the control (group C), "tachyzoite" (group T) and "bradyzoite" (group B) groups were subcutaneously injected in the right flank with 66 µg of ovalbumin, 6.10(6) tachyzoites (125th passage on Vero cells) and 6.10(6) bradyzoites (collected from a natural infected cow) of B. besnoiti, respectively. Clinical follow-up and blood sampling for serological survey and qPCR were performed during 10 weeks until euthanasia. Molecular and immunohistochemistry examination was achieved on 25 samples of tissue per rabbit. Seroconversion occurred in group T without any clinical signs. Rabbits of group B exhibited a febrile condition (temperature above 40 °C from day 8 to day 11 following injection) with positive qPCR in blood. Cysts of B. besnoiti were found on skin samples and organs of rabbits from group B in tissue explored with threshold cycle (Ct) values below 30. These results suggest a higher virulence of bradyzoites in rabbits than Vero cell-cultivated tachyzoites. The proposed model could be used to assess the in vivo effectiveness of vaccine or drugs against cattle besnoitiosis.


Asunto(s)
Coccidiosis/veterinaria , Conejos/parasitología , Sarcocystidae/patogenicidad , Animales , Bovinos , Chlorocebus aethiops , Coccidiosis/parasitología , Europa (Continente) , Femenino , Inmunohistoquímica/veterinaria , Sarcocystidae/fisiología , Células Vero , Virulencia
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