Virulence-associated genes in faecal and clinical Escherichia coli isolates cultured from broiler chickens in Australia.

A healthy chicken's intestinal flora harbours a rich reservoir of Escherichia coli as part of the commensal microbiota. However, some strains, known as avian pathogenic E. coli (APEC), carry specific virulence genes (VGs) that enable them to invade and cause extraintestinal infections such as avian colibacillosis. Although several VG combinations have been identified, the pathogenic mechanisms associated with APEC are ill-defined. The current study screened a subset of 88 E. coli isolates selected from 237 pre-existing isolates obtained from commercial poultry flocks in Australia. The 88 isolates were selected based on their enterobacterial repetitive intergenic consensus (ERIC) and antimicrobial resistance (AMR) profiles and included 29 E. coli isolates cultured from chickens with colibacillosis (referred to as clinical E. coli or CEC) and 59 faecal E. coli (FEC) isolates cultured from clinically healthy chickens. The isolates were screened for the presence of 35 previously reported VGs. Of these, 34 were identified, with iucA not being detected. VGs focG, hlyA and sfa/foc were only detected in FEC isolates. Eight VGs had a prevalence of 90% or above in the CEC isolates. Specifically, astA (100%); feoB (96.6%); iutA, iss, ompT, iroN and hlyF (all 93.1%); and vat (89.7%). The prevalence of these were significantly lower in FEC isolates (astA 79.7%, feoB 77.9%, iutA 52.5%, iss 45.8%, ompT 50.9%, iroN 37.3%, hlyF 50.9% and vat 42.4%). The odds ratios that each of these eight VGs were more likely to be associated with CEC than FEC ranged from 7.8 to 21.9. These eight VGs may be used to better define APEC and diagnostically detect APEC in Australia. Further investigations are needed to identify the roles of these VGs in pathogenicity.

Antimicrobial susceptibility, plasmid replicon typing, phylogenetic grouping, and virulence potential of avian pathogenic and faecal Escherichia coli isolated from meat chickens in Australia.

Globally, avian colibacillosis is a leading cause of morbidity and mortality in poultry, associated with economic losses and welfare problems. Here, clinical avian pathogenic E. coli isolates (CEC; n = 50) and faecal E. coli isolates from healthy (FEC; n = 187) Australian meat chickens collected between 2006 and 2014 were subjected to antimicrobial susceptibility testing, phylogenetic grouping, plasmid replicon (PR) typing, multilocus sequence typing, and virulence gene (VG) profiling. Extended-spectrum cephalosporin (ESC)- and fluoroquinolone (FQ)-resistant E. coli isolates underwent further genetic characterization. Significant proportions of CEC and FEC were, respectively, susceptible (13/50; 48/187) or MDR (9/50; 26/187) to 20 tested antimicrobials. Phylogenetic groups A and C, and PR types IncFIB and IncFrep were most represented. Five tested CEC-associated VGs were more prevalent in CEC (≥ 90%) than FEC (≤ 58%). Some isolates (CEC n = 3; FEC n = 7) were resistant to ESCs and/or FQs and possessed signature mutations in chromosomal FQ target genes and plasmid-mediated qnrS, blaCMY-2, and blaDHA-1 genes. Sequence type 354 (n = 4), associated with extraintestinal infections in a broad range of hosts, was prevalent among ESC- and/or FQ-resistant FEC. This study confirmed existence of a small reservoir of ESC- and FQ-resistant E. coli in Australian commercial meat chickens despite absence of use in the industry of these drugs. Otherwise, diversity of VGs and PR types in both FEC and CEC populations was identified. We hypothesize that the source of ESC- and FQ-resistant E. coli is external to poultry production facilities.RESEARCH HIGHLIGHTSLow-level resistance to older and newer generation antimicrobial drugs detected.The most common sequence type (ST) associated with FQ resistance was ST354 (4/10).A small proportion of CEC (n = 3) and FEC (n = 7) were resistant to ESCs and/or FQs.

Prevalence of Tritrichomonas foetus in beef bulls slaughtered at two abattoirs in northern Australia.

Bovine trichomoniasis, caused by the protozoal parasite Tritrichomonas foetus, is a highly contagious venereal disease characterised by early pregnancy loss, abortion and pyometra. Persistently infected bulls and cows are the primary reservoirs of infection in infected herds. This research investigated the prevalence of T. foetus infection in bulls from properties located across northern Australia and New South Wales. Preputial samples were collected from 606 bulls at slaughter and tested for T. foetus using the VetMAX-Gold Trich Detection Kit (Thermo Fisher Scientific). The apparent prevalence of T. foetus infection varied between regions, with northern regions in the Northern Territory, Queensland and Western Australia showing a prevalence of 15.4%, 13.8% and 11.4%, respectively. There was some evidence of an association between infection and postcode (P = 0.06) and increasing bull age (P = 0.054). This study confirms that T. foetus infection is likely to be present in many beef breeding herds and contributing to lower than expected reproductive performance, particularly across northern Australia.

Root and canal morphology of mandibular first and second molar teeth in a Jordanian population.

AIM: To examine the root and canal morphology of mandibular permanent first and second molar teeth in a Jordanian population. METHODOLOGY: A total of 685 extracted mandibular first and second permanent molar teeth were collected from dental clinics within north Jordan. The teeth were examined visually and the root number and morphology were recorded. After that, access cavities were prepared, pulp tissue dissolved by sodium hypochlorite and the root canals injected with Indian ink. Stained teeth were decalcified with 10% nitric acid, dehydrated with ascending concentrations of alcohol and rendered clear by immersion in methyl salicylate. The following observations were evaluated: (i) number of canals per root; (ii) number of root canals per tooth; (iii) canal configuration in each root; (iv) number and location of lateral canals and (v) presence of intercanal communications. RESULTS: Of 330 mandibular first molars, the majority had three (48%) or four (46%) canals, whilst 4% had a third disto-lingual root. Of 355 mandibular second molars, 58% had three canals, 19% two and 17% had four canals, whilst 10% had C-shaped roots. The most prevalent canal configuration in the mesial root of both first (53%) and second (40%) molars was type IV, and in distal roots was type I (54% in first and 79% in second molars). CONCLUSIONS: Jordanian mandibular first and second molar teeth exhibit features close to the average Caucasian root and canal morphology.

Root form and canal morphology of mandibular premolars in a Jordanian population.

AIM: To investigate the root canal anatomy of mandibular premolars in a Jordanian population. METHODOLOGY: Nine hundred extracted mandibular premolars were examined. After the length of the teeth was measured, the presence of developmental grooves and furcated roots was noted. Following the preparation of access cavities, pulp tissue was removed and the canal systems were stained. The teeth were then rendered clear by demineralization and immersion in methyl salicylate. Cleared teeth were examined and the following features were evaluated: (i) type of root canals; (ii) presence and location of lateral canals; transverse anastomosis; (iii) location of apical foramina; and (iv) frequency of apical deltas. RESULTS: The mean lengths of first and second mandibular premolars were 22.6 mm (18-27.5 mm) and 22.2 mm (16-26.5 mm), respectively. Although the majority of the specimens corresponded to Vertucci's classification scheme, analysis of this large data set revealed four additional root canal morphologies. Variable root canal morphologies were found in the mandibular first premolars; two separate apical foramina were found in 33% of the teeth with two canals, compared to 6.2% with one apical foramen. Teeth with three separate apical foramina were scarce (2.2%). The majority of the mandibular second premolars had a single canal; 72% of teeth possessed type I canal systems, whilst 22.8% of the roots had two canals with two separate apical foramina. CONCLUSIONS: The prevalence of multiple canals in the investigated Jordanian mandibular premolars was high, especially for the second mandibular premolar, in comparison with previous studies performed on populations of different racial origin.

Root canal morphology of mandibular incisors in a Jordanian population.

AIM: To investigate the root canal morphology of mandibular incisors in a Jordanian population using a canal staining and tooth-clearing technique. METHODOLOGY: Four hundred and fifty extracted mandibular incisors were collected from dental clinics within north Jordan. Following pulp tissue removal and staining of the canal systems, the teeth were decalcified with 5% nitric acid, dehydrated with ascending concentrations of alcohol and rendered clear by immersion in methyl salicylate. Cleared teeth were examined by eye and the following features were evaluated: (i) number and type of root canals; (ii) presence and location of lateral canals and intercanal communications; (iii) location of apical foramina; and (iv) frequency of apical deltas. RESULTS: The majority of mandibular incisors had a single canal (73.8% of teeth possessed a Type I canal system). Although 26.2% of the roots possessed two canals, only 8.7% had two separate apical foramina. CONCLUSIONS: The prevalence of two canals in this group of mandibular incisors was 26.2% and is within the range of previous studies performed on populations of different racial origin.

Quantitative analysis of substance P, neurokinin A and calcitonin gene-related peptide in pulp tissue from painful and healthy human teeth.

AIM: The purpose of this study was to investigate the levels of substance P (SP), neurokinin A (NKA) and calcitonin gene-related peptide (CGRP) in painful and healthy human dental pulps. METHODOLOGY: Forty-six samples of pulp tissue were collected from extracted or endodontically treated painful teeth and 20 from clinically healthy teeth extracted for orthodontic reasons. All pulp samples were boiled in 0.5 m acetic acid for 10 min, centrifuged and the supernatant collected. SP, NKA and CGRP levels were measured using radioimmunoassay. RESULTS: Substance P and CGRP were present in all samples and NKA was detected in 96% of the pulps. CGRP was present in much higher concentrations than SP and NKA in both painful and non-painful teeth. The painful teeth had significantly higher concentrations of SP (P = 0.02), NKA (P < 0.001) and CGRP (P = 0.03) than non-painful teeth. The concentration of CGRP was significantly higher in the pulps of smokers compared with non-smokers (P = 0.02). CONCLUSIONS: Elevated levels of these neuropeptides in pulps from painful teeth indicate that they may play an important role in the process of pulpal inflammation and pain. Further investigation of the association between these neuropeptides and pulpal status may help to improve our understanding of pulpal inflammation and dental pain.

A comparison of four extraction methods for substance P, neurokinin A and calcitonin gene-related peptide from human dental pulp tissue.

Measuring neuropeptides in biological tissues by radioimmunoassay requires efficient extraction that maintains their immunoreactivity. Many different methods for extraction have been described, but there is little information on optimal extraction methods for individual neuropeptides from human dental pulp tissue. The aim was therefore to identify an effective extraction procedure for three pulpal neuropeptides; substance P, neurokinin A and calcitonin gene-related peptide. Tissue was obtained from 20 pulps taken from teeth freshly extracted for orthodontic reasons. The pulp samples were divided into four equal groups and different extraction methods were used for each group. Boiling whole pulp in acetic acid gave the highest overall yield and, in addition, offered an easy and rapid means of pulp tissue processing. The use of protease inhibitors did not increase the recovery of the immunoreactive neuropeptides but did provide the best combination of maximal recoveries and minimal variability. These results should be useful for planning the extraction of these neuropeptides from human pulp tissue in future studies.

The effect of a resin-modified glass ionomer restorative material on artificially demineralised dentine caries in vitro.

OBJECTIVES: Recurrent caries is one of the most common reasons for the replacement of restorations. One method of reducing the frequency of this problem may be by using fluoride-releasing restorative materials. The aim of this in vitro study was to evaluate the progression of artificial lesions around resin-modified glass ionomer (Vitremer) restorations placed in root surfaces. Class V cavities were prepared on the mesial and distal surfaces of 28 human premolar teeth. Artificial carious wall lesions were created in all cavities. The root of each tooth was then hemisected through the middle of the two cavities, before being restored with either the glass ionomer or amalgam, while the opposing cavities on the same root portion were varnished as negative controls. Forty-eight specimens were pH-cycled in remineralising and demineralising solutions for 20 h and 4 h, respectively, each day for 4 weeks, whilst in a smaller control group of eight specimens, the demineralising solution was replaced with deionised water. Mineral changes in the carious lesions were evaluated using contact microradiography. RESULTS: Results showed that varying degrees of subsurface demineralisation and remineralisation were evident, with a laminated appearance in lesions adjacent to the glass ionomer. There was higher remineralisation in the glass ionomer-filled cavities compared with the amalgam-filled cavities. In the water-cycled group, glass ionomer showed an increase in mineral content (p < 0.05), while no changes were observed in the amalgam-filled cavities. CONCLUSIONS: This model has shown that glass ionomer has a greater potential than amalgam for remineralisation of artificially created wall lesions within an acidic environment.