Comparative genomic fingerprinting of Campylobacter: application in routine public health surveillance and epidemiological investigations.

A subtyping methodology for Campylobacter, Comparative Genomic Fingerprinting (CGF40), has been described recently. The objective of this study was to assess the utility of CGF40 as a tool to enhance routine public health surveillance of campylobacteriosis. Isolates of Campylobacter from across the province were requested and sent for CGF40 subtyping. Epidemiological data from cases reported to public health officials in Nova Scotia, Canada, from January 2012 to March 2015 were linked with blinded CGF40 subtyping results. CGF40 was epidemiologically valid; subtyping discerned known epidemiologically related isolates and augmented case-finding. Predominant sources and locations of subtype detection from the national reference database showed some study subtypes were rare and even novel to the database, while others were more commonly identified over multiple years and with exposures locally and internationally. A case-case study design was applied to examine risk factors for the most common CGF40 subtypes detected. Differences in the epidemiology of different CGF40 subtypes were observed. Statistically significant associations were noted for specific subtypes with rural residence, local exposure, contact with a pet dog or cat, contact with chickens, and drinking unpasteurized milk. With prospective use, CGF40 could potentially identify unrecognized outbreaks and contribute to epidemiological investigations of case clusters.

Heterogeneity in the spatial distribution of bacterial sexually transmitted infections.

OBJECTIVES: Detailed knowledge of the spatial distribution of disease is required to inform service delivery and plan effective interventions. In order to elucidate the spatial epidemiology of three common sexually transmitted infections (STI) further, any significant spatiotemporal clustering of gonorrhoea, chlamydia or syphilis cases in New South Wales (NSW) was detected and described. METHODS: Eleven years of notified STI case data were analysed. Calculation of age and sex-stratified incidence rates was followed by spatiotemporal cluster analyses to investigate differences in the epidemiology of gonorrhoea, chlamydia and syphilis. RESULTS: More than one-third of all gonorrhoea, chlamydia and syphilis cases in NSW were detected within cluster areas. Gonorrhoea cases were the most highly clustered, followed by syphilis, then chlamydia. Clusters were highly significant and relative risk estimates ranged from 1.6 to 22.9. CONCLUSION: The findings establish the high degree of geographical heterogeneity in STI incidence in NSW and indicate that the postal area of residence is an important predictor of STI incidence. Geographical surveillance could be incorporated into routine STI surveillance to identify populations in need of intervention. The evidence presented in this report indicates a need to implement geography-specific and phase-appropriate STI prevention and control strategies.

Calcium-stimulated short-circuit currents in the canine proximal colonic epithelium: effects of DK-PGD2, a metabolite of prostaglandin D2.

Prostaglandin D2 (PGD2) has marked inhibitory effects on the canine proximal colonic epithelium set up in Ussing chambers. These effects involved a receptor that is pharmacologically distinct from the classical DP, presumably the recently identified CRTH2/DP2 variety. The mechanism underlying these effects was studied using 13,14-dihydro-15-keto-PGD2 (DK-PGD2), a stable metabolite of the parent prostanoid. The metabolite quickly reversed short circuit currents (I(sc)) stimulated by diverse agonists. Greater inhibitory effects were seen with stimulants such as carbachol and cyclopiazonic acid (CPA) rather than with forskolin or protein kinase A activators. Since the same stimulants were differentially affected by removal and replacement of serosal Ca2+, we tested the possibility that the prostanoid inhibited basolateral Ca2+ entry. In the absence of serosal Ca2+, tissues primed with CPA demonstrated concentration-dependent increases in I(sc), to cumulative additions of Ca2+ or Sr2+, though the former was more potent. Cl- removal and pretreatment with bumetanide virtually abolished responses, suggesting that the increase in I(sc) reflected Ca2+ dependent Cl- secretion. Though responses were insensitive to the L-type channel antagonist, verapamil, a marked inhibition was seen in the presence of metal cations (Gd3+, Cd2+, and La3+). Pretreatment with DK-PGD2 inhibited responses to Ca2+ in CPA-primed tissues. Thus, basolateral Ca2+ entry via store-operated Ca2+ channels may be the locus for the inhibitory effects of PGD2 in this tissue. These results could indicate a potential transduction mechanism for the novel DP receptor variously called CRTH2 or DP2.

Protease-activated receptor-1 stimulates Ca(2+)-dependent Cl(-) secretion in human intestinal epithelial cells.

The thrombin receptor, protease-activated receptor-1 (PAR-1), has wide tissue distribution and is involved in many physiological functions. Because thrombin is in the intestinal lumen and mucosa during inflammation, we sought to determine PAR-1 expression and function in human intestinal epithelial cells. RT-PCR showed PAR-1 mRNA expression in SCBN cells, a nontransformed duodenal epithelial cell line. Confluent SCBN monolayers mounted in Ussing chambers responded to PAR-1 activation with a Cl(-)-dependent increase in short-circuit current. The secretory effect was blocked by BaCl2 and the Ca(2+)-ATPase inhibitor thapsigargin, but not by the L-type Ca(2+) channel blocker verapamil or DIDS, the nonselective inhibitor of Ca(2+)-dependent Cl(-) transport. Responses to thrombin and PAR-1-activating peptides exhibited auto- and crossdesensitization. Fura 2-loaded SCBN cells had increased fluorescence after PAR-1 activation, indicating increased intracellular Ca(2+). RT-PCR showed that SCBN cells expressed mRNA for the cystic fibrosis transmembrane conductance regulator (CFTR) and hypotonicity-activated Cl(-) channel-2 but not for the Ca(2+)-dependent Cl(-) channel-1. PAR-1 activation failed to increase intracellular cAMP, suggesting that the CFTR channel is not involved in the Cl(-) secretory response. Our data demonstrate that PAR-1 is expressed on human intestinal epithelial cells and regulates a novel Ca(2+)-dependent Cl(-) secretory pathway. This may be of clinical significance in inflammatory intestinal diseases with elevated thrombin levels.