g Highly virulent pantropic canine coronavirus (CCoV) strains belonging to subtype IIa were recently identified in dogs. To assess the distribution of such strains in Europe, tissue samples were collected from 354 dogs that had died after displaying systemic disease in France (n ‫؍‬ 92), Hungary (n ‫؍‬ 75), Italy (n ‫؍‬ 69), Greece (n ‫؍‬ 87), The Netherlands (n ‫؍‬ 27), Belgium (n ‫؍‬ 4), and Bulgaria (n ‫؍‬ 1). A total of 124 animals tested positive for CCoV, with 33 of them displaying the virus in extraintestinal tissues. Twenty-four CCoV strains (19.35% of the CCoV-positive dogs) detected in internal organs were characterized as subtype IIa and consequently assumed to be pantropic CCoVs. Sequence and phylogenetic analyses of the 5= end of the spike protein gene showed that pantropic CCoV strains are closely related to each other, with the exception of two divergent French viruses that clustered with enteric strains. C oronaviruses (CoVs; order Nidovirales, family Coronaviridae) are positive-sense RNA viruses commonly associated with mild infections in birds and mammals. The family Coronaviridae is now organized into two subfamilies, Coronavirinae and Torovirinae, with the former including four different genera, Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus. Canine coronavirus (CCoV) belongs to the genus Alphacoronavirus and forms a unique species, Alphacoronavirus 1, along with feline coronaviruses (FCoVs), transmissible gastroenteritis virus of swine (TGEV), and its derivative, porcine respiratory coronavirus (PRCoV) (1, 2).CCoVs are paradigmatic of the CoV genetic evolution and complexity (3, 4). In addition to the known genotypes, CCoV types I (CCoV-I) and II (CCoV-II) (5), which share up to 96% of nucleotide sequence identity in the viral genome but are highly divergent in the spike (S) protein gene (6), CCoV subtypes and biotypes have been more recently identified (3, 4). Detection of TGEV/CCoV recombinant strains led to the classification of CCoV-II into two subtypes, including the classical (CCoV-IIa) and recombinant (CCoV-IIb) subtypes, respectively (7,8). A hypervirulent CCoV-IIa strain, designated pantropic CCoV, was isolated from dead pups at a pet shop in Italy in 2005 (9). The virus, strain CB/05, was associated with severe clinical signs and postmortem lesions. Experimental infections of dogs reproduced the disease, with the severity varying with the age and immune status of the infected animals (10, 11) but invariably leading to long-term lymphopenia (12).Natural outbreaks of pantropic CCoV infection have been reported in France and Belgium (13), Greece (14), and Italy (15).The aim of the present study is to report the detection of pantropic CCoV in some European countries. MATERIALS AND METHODSSample collection. A total of 354 carcasses of dogs that died after displaying systemic disease consisting of fever, leukopenia, depression, enteritis, respiratory distress, and/or neurological signs were sampled from 2009 to 2011 (Table 1). Cases were admitted to the study i...

We report the genetic and biological characterisation of a novel pantropic canine coronavirus (CCoV), strain 450/07, which caused the death of a 60-day-old miniature pinscher. At the genetic level, this virus was strictly related to the prototype strain CB/05, but displayed some unique features. After experimental infection with the new pantropic isolate, most inoculated dogs showed diarrhoea and acute lymphopenia. Gross lesions and histological changes were mainly evident in the gut and lymphoid tissues, although some animals showed remarkable changed also in parenchymatous organs. The viral RNA was detected in the faeces and/or internal organs of most pups. These findings seem to indicate that strain 450/07 is able to spread to internal organs (mainly lymphoid tissues), causing lymphopenia but inducing a mild disease.

Background: Lung, kidney and small intestine are involved in fetal volume regulation and amniotic fluid secretion and play a pivotal role in the transition from intrauterine to extrauterine life. Objective: This study was performed to determine the ontogeny of mineralocorticoid receptors (MR) and glucocorticoid receptors (GR), and of MR- and GR-regulated genes and proteins, serum and glucocorticoid-induced kinase (Sgk-1), epithelial sodium channel (ENaCα), and Na,K-ATPase α1. Methods: Lung, renal cortex and medulla, and small intestine were collected from fetuses at 80, 100, 120, 130 and 145 days’ gestation and from day 1 and 7 neonatal lambs. Real-time PCR was performed to determine mRNA concentration for MR, GR, the 11β-hydroxysteroid dehydrogenases (11β-HSD1 and 2), Sgk-1, ENaCα, and Na,K-ATPase α1. Protein expression of ENaCα and Na,K-ATPase α1 in whole cell and membrane fractions was determined by immunoblotting. Results: Expression of corticosteroid-induced genes in renal cortex increases at term; in small intestine the induction occurs postnatally. In contrast, in lung expression of MR and GR mRNAs were greater at 100 days to term than postnatally and 11β-HSD1 peaked at 145 days; the corticosteroid-induced genes also increased prenatally: Sgk-1 and ENaCα increased by 120 days, peaking at 145 days, and Na,K-ATPase α1 was greatest at 130 days. Conclusions: The expression of high levels of MR and 11β-HSD1 in preterm fetal lung suggest low endogenous fetal cortisol may exert actions at the high affinity MR in vivo, leading to increases in expression of sodium channels important in the regulation of lung liquid secretion and reabsorption.

The pattern of Fos-like immunoreactivity (FLI) in the periaqueductal gray (PAG) associated with activation of arterial chemoreceptors versus baroreceptor afferents was examined in urethane-anesthetized rats. Chemoreflex responses elicited by repeat intravenous injections of potassium cyanide (KCN; 90 microg/kg) significantly increased FLI in all columns of the PAG relative to saline-injected animals. Pressor responses elicited by intravenous phenylephrine (PE) produced a similar pattern of increased FLI throughout the PAG except in the dorsomedial and lateral columns of the caudal PAG, where FLI was minimal. Chemoreflex responses were unaltered by blockade of excitatory amino acid receptors in the dorsomedial PAG, and < 10% of the neurons of the caudal PAG that expressed FLI after KCN stimulation were retrogradely labeled from the A5 region of the caudal ventrolateral pons. These results indicate that integration of chemoreceptor inputs occurs primarily in the dorsal and lateral columns of the caudal PAG, but these neurons have little direct descending influence over lower brain stem regions integral to the central arterial chemoreflex arc.

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Restraint stress ulcers in rats were developed and procedures evaluated designed at preservation of intact gastric microcirculation (pO2). Neither prior truncal vagotomy, splanchnicotomy nor combined dissection of abdominal autonomic nerves were effective in preventing the stress mediated fall of mucosal pO2 and the rise in plasma glucagon. The ulcer index remained elevated and gastrin essentially was unchanged. Prophylactic injection of increasing doses (1, 2, 4, 8, 16 U/kg secretin maintained microcirculation at pO2-levels subnormal for unstressed animals (Vmax 15.53 mm Hg; Km. 0.99 U/kg), but simultaneously brought about a continuous rise in serum gastrin. Up to 8 U/kg plasma glucagon was higher than in saline control groups reaching a peak value with 2 U/kg when ucler index showed its nadir. Secretin therapy (4, 8 U/kg) markedly improves both mucosal pO2 and ulcer index. It is suggested that breakdown of gastric microcirculation may not be solely responsible for stress ulcer development.

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