The catabolite control protein CcpA is a transcriptional regulator conserved in many Gram-positives, controlling the efficiency of glucose metabolism. Here we studied the role of Bacillus cereus ATCC 14579 CcpA in regulation of metabolic pathways and expression of enterotoxin genes by comparative transcriptome analysis of the wild-type and a ccpA-deletion strain. Comparative analysis revealed the growth performance and glucose consumption rates to be lower in the B. cereus ATCC 14579 ccpA deletion strain than in the wild-type. In exponentially grown cells, the expression of glycolytic genes, including a non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase that mediates conversion of D-glyceraldehyde 3-phosphate to 3-phospho-D-glycerate in one single step, was down-regulated and expression of gluconeogenic genes and genes encoding the citric acid cycle was up-regulated in the B. cereus ccpA deletion strain. Furthermore, putative CRE-sites, that act as binding sites for CcpA, were identified to be present for these genes. These results indicate CcpA to be involved in the regulation of glucose metabolism, thereby optimizing the efficiency of glucose catabolism. Other genes of which the expression was affected by ccpA deletion and for which putative CRE-sites could be identified, included genes with an annotated function in the catabolism of ribose, histidine and possibly fucose/arabinose and aspartate. Notably, expression of the operons encoding non-hemolytic enterotoxin (Nhe) and hemolytic enterotoxin (Hbl) was affected by ccpA deletion, and putative CRE-sites were identified, which suggests catabolite repression of the enterotoxin operons to be CcpA-dependent. The catabolite control protein CcpA in B. cereus ATCC 14579 is involved in optimizing the catabolism of glucose with concomitant repression of gluconeogenesis and alternative metabolic pathways. Furthermore, the results point to metabolic control of enterotoxin gene expression and suggest that CcpA-mediated glucose sensing provides an additional mode of control in moderating the expression of the nhe and hbl operons in B. cereus ATCC 14579.
The antimetabolite mangotoxin is a key factor in virulence of Pseudomonas syringae pv. syringae strains which cause apical necrosis of mango trees. Previous studies showed that mangotoxin biosynthesis is governed by the mbo operon. Random mutagenesis led to the identification of two other gene clusters that affect mangotoxin biosynthesis. These are the gacS/gacA genes and mgo operon which harbors the four genes mgoBCAD.
The aim of our study was to investigate the virulence and resistance of STEC from small ruminants farms in the Netherlands. Moreover, the potential transmission of STEC between animals and humans on farms was evaluated. From 182 farms, in total 287 unique STEC isolates were successfully recovered from animal samples. In addition, STEC was isolated from eight out of 144 human samples. The most detected serotype was O146:H21, however, among other serotypes also O26:H11, O157:H7 and O182:H25 isolates were present. Whole genome sequencing covering all human isolates and 50 of the animal isolates revealed a diversity of stx1, stx2, and eae sub-types and an additional 57 virulence factors. The assessed antimicrobial resistance phenotype, as determined by microdilution, was concordant with the genetic profiles identified by WGS. WGS also showed that three of the human isolates could be linked to an animal isolate from the same farm. The obtained STEC isolates showed great diversity in serotype, virulence and resistance factors. Further analysis by WGS allowed for an in-depth assessment of virulence and resistance factors present and to determine the relatedness of human and animal isolates.
Listeria monocytogenes (Lm) is ubiquitous in nature and known for its ability to contaminate foods during production processes. Near real-time monitoring of whole genome sequences from food and human isolates, complemented with epidemiological data, has been used in the Netherlands since 2019 to increase the speed and success rate of source finding in the case of (active) clusters. Nine clusters with 4 to 19 human cases investigated between January 2019 and May 2023 are described. Fish production sites were most often linked to outbreaks of listeriosis (six clusters), though other types of food businesses can face similar Lm problems, as the production processes and procedures determine risk. The results showed that low levels of Lm in food samples can still be linked to disease. Therefore, the investigation of a cluster of cases and deployment of the precautionary principle helps to focus on safe food and to prevent further cases. Good practice of environmental monitoring within a food business allows early detection of potential issues with food safety and helps food businesses to take appropriate measures such as cleaning to prevent regrowth of Lm and thus future outbreaks.
Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen associated with illness ranging from mild diarrhoea to haemolytic uremic syndrome (HUS) or even death. Cross-sectoral data sharing provides an opportunity to gain insight in reservoirs and sources of human infections and starting points for pro-active measures. Nevertheless, phylogenetic clustering of STEC strains from animals, food and human cases is low in the Dutch surveillance system. This is partly due to the substantial contribution of international travel and person-to-person spread in the STEC epidemiology. Furthermore, some STEC strains causing disease in humans may have a human reservoir. Although the main reservoirs and sources are included in the Dutch monitoring programmes, some animals and food products may be under-recognised as potential sources of human infections. More effort in investigating the role of other reservoirs beyond the well-known can provide a better understanding on STEC ecology in general, improving surveillance and source attribution, and ultimately provide better guidance for monitoring and source finding. This also implies having good diagnostics in place and isolates available for typing. Therefore, on the human side of the surveillance, the decision has been made to start isolating STEC at national level.