Multiplex Lateral Flow Immunoassay for the Detection of Expanded-Spectrum Hydrolysis and CTX-M Enzymes

Background: Early detection of expanded-spectrum cephalosporinase (ESC) hydrolyzing ß-lactamases is essential for antibiotic stewardship. Here we have developed a multiplex lateral flow immunoassay (LFIA) that detects cefotaxime-hydrolyzing activity as well as the most prevalent ESC-hydrolyzing ß-lactamases: the CTX-M-like. Methods: The Rapid LFIA ESC test was evaluated retrospectively on 188 (139 Enterobacterales, 30 Pseudomonas spp. and 14 Acinetobacter spp.) agar-grown bacterial isolates with well-characterized ß-lactamase content. One single colony was resuspended in 150 µL extraction buffer containing cefotaxime, incubated at room temperature for 30 min prior to loading on the LFIA for reading within 10 min. Results: Out of the 188 isolates, all 17 that did not express a β-lactamase hydrolyzing cefotaxime gave negative results, and all 171 isolates expressing a β-lactamase known to hydrolyze cefotaxime, gave a positive test result. In addition, all 86 isolates expressing a CTX-M-variant belonging to one of the five CTX-M-subgroups were correctly identified. The sensitivity and specificity was 100% for both tests. Conclusions: The results showed that the multiplex LFIA was efficient, fast, low cost and easy to implement in routine laboratory work for the confirmation of ESC hydrolyzing activity and the presence of CTX-M enzymes.

Substantial variability in morphological scaling among bumblebee colonies

Differences in organ scaling among individuals may play an important role in determining behavioural variation. In social insects, there are well-documented intraspecific differences in colony behaviour, but the extent that organ scaling differs within and between colonies remains unclear. Using 12 different colonies of the bumblebee Bombus terrestris , we aim to address this knowledge gap by measuring the scaling relationships between three different organs (compound eyes, wings and antennae) and body size in workers . Though colonies were exposed to different rearing temperatures, this environmental variability did not explain the differences of the scaling relationships. Two colonies had differences in wing versus antenna slopes, three colonies showed differences in wing versus eye slopes and a single colony has differences between eye versus antenna slopes. There are also differences in antennae scaling slopes between three different colonies, and we present evidence for putative trade-offs in morphological investment. We discuss the utility of having variable scaling among colonies and the implication for understanding variability in colony fitness and behaviour.

Prevalence of non-O157 Shiga Toxin-producing E. Coli in Children and Calves in Al- Muthanna Province, Iraq

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.

Bacterial genomic epidemiology with mixed samples

Genomic epidemiology is a tool for tracing transmission of pathogens based on whole-genome sequencing. We introduce the mGEMS pipeline for genomic epidemiology with plate sweeps representing mixed samples of a target pathogen, opening the possibility to sequence all colonies on selective plates with a single DNA extraction and sequencing step. The pipeline includes the novel mGEMS read binner for probabilistic assignments of sequencing reads, and the scalable pseudoaligner Themisto. We demonstrate the effectiveness of our approach using closely related samples in a nosocomial setting, obtaining results that are comparable to those based on single-colony picks. Our results lend firm support to more widespread consideration of genomic epidemiology with mixed infection samples.

Listeria monocytogenes sensitivity to antimicrobial treatments depends on cell origin

In this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment (mild heating combined with terpenoids) in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase. After treatment, culturability on BHI medium and viability assessed by flow cytometry were evaluated. Our results showed that the cell origin significantly impacted viability and culturability of L. monocytogenes towards antimicrobial treatment. The mild heat treatment combined or not with terpenoids mainly affected culturability rather than viability, although the culturability of cells from single colony was less impacted. Therefore, to mimic the worst scenario, these latter were selected to contaminate Gorgonzola rind and roast beef slices and we evaluated the ability of L. monocytogenes cells to recover their culturability (on ALOA agar medium) and to growth on the food matrix stored at 4 °C for 7 days. Our results suggest that only Gorgonzola rind allowed a partial recovery of the culturability of cells previously heated in presence or not of terpens. In conclusion, we found a connection between the cell history and sensitivity toward an antimicrobial treatment, underlying the importance to standardize the experimental procedures (starting from the cells to be used in the assay) in the assessment of cell sensitivity to a specific treatment. Finally, our study clearly indicated that VBNC cells can resuscitate under favorable conditions on a food matrix, becoming a threat for consumer’s health.

A simulation to identify the optimal area to survey in the early stage of invasion by Solenopsis invicta

Efforts to eradicate invasive alien species commonly use simulations to calculate the cost-effectiveness of surveys. Although eradication of Solenopsis invicta in the early stages of an invasion is important, few simulations are available to calculate the cost-effectiveness of surveys when a single colony has been detected. In the case of S. invicta, it is difficult to determine from the status of the detected colony whether new queens have dispersed, so it is necessary to consider dispersal as a probabilistic event and calculate its probability. We therefore first constructed a mathematical model in which we used Bayesian statistics to estimate the probability of dispersal as a function of the results of the survey. This mathematical model revealed that the efficacy of the survey and the associated cost differed greatly between cases depending on whether dispersal was or was not confirmed. Next, we developed a simulation that incorporated this mathematical model to inform the determination of the survey area when a single colony had been detected. The simulation showed how ecological parameters and geographical information could be used to identify an efficacious survey area, even in heterogeneous landscapes such as international ports where invasions occur sporadically. Finally, we used this simulation to assess the efficacy of a survey in the case of an S. invicta outbreak at the Port of Tokyo, Japan. The results suggested that the survey covered a sufficiently wide area but that it could have been designed in a more efficacious manner.

A redescription of Stenopora and the type species Stenopora tasmaniensis Lonsdale, 1844 (Trepostomata, Bryozoa)

Type material for Stenopora tasmaniensis Lonsdale, 1844 was lost in the late nineteenth century, and subsequent descriptions of the genus have been based on material incorrectly assigned to the type species. A neotype is erected for S. tasmaniensis from the original type locality and the genus redescribed. The genus exhibits ramose, frondescent, encrusting, and massive colony morphologies, diaphragms are absent, and acanthostyles of a single size surround each aperture. This single size of acanthostyles aligns with the original type species description; however, it differs from the subsequently accepted genus description and may result in existing species being removed from the genus. Analysis of zooecial characters of a single colony exhibiting both frondescent and ramose morphologies reveals statistically significant differences between subsampled sections, despite being from the same colony. Differences relate to details of zooecial parameters and are not controlled by colony morphology. This variation within a single colony confirms the importance of using qualitative characters alongside quantitative measures in defining Paleozoic bryozoan species.