How to to identify Bacillus cereus group with MALDI TOF MS

What is Bacillus cereus group?

The Bacillus cereus group, also known as B. cereus sensu lato, is a subdivision of the Bacillus genus that comprises several closely related species of Gram-positive, rod-shaped, spore-forming bacteria. The group includes eight formally recognized species: B. cereus sensu stricto, B. anthracis, B. thuringiensis, B. weihenstephanensis/B. mycoides, B. pseudomycoides, B. cytotoxicus, and B. toyonensis.

These bacteria are widespread in nature and are commonly associated with food poisoning, intestinal infections, and other pathogenic potential. The B. cereus group has been the subject of extensive research, and its taxonomy and species status have been determined based on phenotypic characteristics and genotypic methods, including Whole Genome Sequencing. The bacteria within this group have a wide range of virulence factors and can cause foodborne illness, localized wound and eye infections, as well as systemic disease.

Additionally, certain strains of B. thuringiensis are entomopathogens and have been commercialized for use as biopesticides, while some strains have been reported to cause infection in immunocompromised individuals.

The phylogeny of the B. cereus group is based on chromosomal genes, and the assignment of a specific isolate to a single species or phylogenetic group can be complicated due to the presence of important virulence and phenotypic traits that are plasmid-borne and can be lost or horizontally transferred.

In summary, the Bacillus cereus group encompasses a diverse set of bacteria with pathogenic potential and has implications for food safety, public health, and microbiology research.

Commercial MALDI-TOF MS systems can only differentiate species of the Bacillus cereus group with difficulty, often only at the group level.
However, the adapted work-up protocols and the Mabriteccentral.com algorithm enable a clear differentiation of all B. cereus group species using MALDI-TOF MS.

How we identify Bacillus cereus group with MALDI TOF MS

Fresh sample

Short incubation time (3-5 hours) to minimise the formation of spores

Sample preparation

Washing of sample and physical distruption with glass beads

Spectra acquisition

Sinapic acid as matrix in a massrange from 4-30kDa

Mabriteccentral.com

Identification and interpretation by mabriteccentral and it’s algorithm.

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Nocardia fluminea identified with MALDI TOF MS

Nocardia fluminea

Nocardia fluminea is a species of bacteria belonging to the genus Nocardia. It was described in 2000 as part of the Nocardia salmonicida clade, which also includes Nocardia cummidelens and Nocardia soli. Nocardia is a genus of weakly staining Gram-positive, catalase-positive, rod-shaped bacteria, and some species are known to cause nocardiosis in humans, particularly in immunocompromised individuals. The clinical and laboratory features of Nocardia species, including N. fluminea, are important for understanding their pathogenic potential and for guiding treatment decisions.

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Sphingobium limneticum identified with MALDI TOF MS

Sphingobium limneticum: A Scientific and Humoristic Overview

The Scientific Side

*Sphingobium limneticum* is a fascinating character in the microbial world, making its home in the serene environments of alpine and pre-alpine lakes. This bacterium, a chemo-organoheterotroph, thrives in freshwater ecosystems, showcasing its adaptability and resilience. It’s a member of the *Sphingomonadaceae* family, known for its gram-negative, rod-shaped, motile cells that form yellow, circular, convex colonies on various agar media.

Optimally growing at temperatures between 10 and 40°C (with a sweet spot at 28°C) and pH values from 5 to 10 (preferring a neutral pH 7), *S. limneticum* is not just any microbe; it’s a testament to life’s ability to flourish in diverse conditions. Its cellular machinery includes Q-10 as the dominant quinone, sphingoglycolipids, and 2-hydroxymyristic acid, elements that hint at its complex biochemistry and ecological roles.

The Humoristic Twist

Imagine if *Sphingobium limneticum* had a dating profile. It would probably say: “Thriving single bacterium, loves long swims in alpine lakes. Enjoys a pH-balanced lifestyle and warm summers at 28°C. Looking for a colony to form beautiful, yellow, convex relationships. Must appreciate the finer things in life, like Q-10 and sphingoglycolipids.”

But don’t let its preference for pristine waters fool you; *S. limneticum* is more than just a pretty face in the microbial community. It’s part of the *Sphingomonadaceae* family, a group known for breaking down complex organic compounds. So, it’s not just lounging around in those alpine lakes; it’s cleaning them up, one aromatic compound at a time.

In Conclusion

*Sphingobium limneticum* is a remarkable microbe, showcasing the incredible adaptability and ecological importance of bacteria in freshwater ecosystems. Its ability to thrive in a range of conditions, combined with its biochemical prowess, makes it a key player in its habitat. And while it might not actually be on the microbial dating scene, its role in nature is undoubtedly as crucial as it is fascinating. So, the next time you’re enjoying the serene beauty of an alpine lake, remember that *Sphingobium limneticum* might be silently working below the surface, keeping the waters clean and balanced.

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Pigmentiphaga kullae identified with MALDI TOF MS

Pigmentiphaga kullae

The Azo-Dye Assassin and Party Animal of the Microbial World

Imagine a world where bacteria are the unsung heroes, quietly working behind the scenes to clean up our messes. Enter *Pigmentiphaga kullae*, a bacterium so cool, it doesn’t just survive in the harsh, color-splashed wastelands of dye wastewater; it thrives by eating the very thing that makes these places inhospitable: azo dyes. This microbe is like the eco-friendly cleaner of the bacterial world, taking what we consider waste and turning it into an all-you-can-eat buffet.

But *P. kullae* isn’t just about keeping things clean. It’s also about living life to the fullest. Found in diverse environments, from the dye-infested waters to the serene soils, it’s the social butterfly of the microbial realm. It’s as if *P. kullae* is saying, “Give me your tired, your poor, your huddled masses of azo dyes yearning to be broken down, and I will give you cleanliness.”

And let’s not forget its genome, a treasure trove of potential, with a single circular chromosome packed with over 5,300 predicted coding sequences. This isn’t just any genome; it’s a blueprint for survival and versatility. With genes for nonribosomal peptide synthetase and bacteriocin biosynthesis, but lacking clusters for β-lactones found in other *Pigmentiphaga* genomes, *P. kullae* is like the MacGyver of bacteria, ready to tackle any challenge with a paperclip and a piece of gum.

But what truly sets *P. kullae* apart is its ability to decolorize azo dyes aerobically. In a world where synthetic dyes pollute our waters, *P. kullae* steps up as the eco-warrior, breaking down these complex molecules and reducing environmental damage. It’s not just cleaning up; it’s performing alchemy, transforming pollutants into harmless substances.

So, the next time you see a brightly colored piece of fabric, remember the unseen heroes like *Pigmentiphaga kullae*. They’re not just surviving in a world of waste; they’re thriving, partying, and making the world a cleaner place, one azo dye at a time. *P. kullae*: not all heroes wear capes; some just have a really cool genome and an appetite for pollution.

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Nocardia fluminea identified with MALDI TOF MS

Clostridium sulfidigenes

Clostridium sulfidigenes, a bacterium that could be the life of the microbial party, is a bit of a metalhead, showing a surprising fondness for heavy metals like cadmium and lead. This tiny organism, which could be mistaken for a minuscule alchemist, is not only resistant to these toxic substances but also has a penchant for reducing thiosulfate and sulfur, making it a bit of a chemical whiz in the microbial world.

While it’s not the type to make headlines for cellulose degradation like some of its bacterial brethren, C. sulfidigenes has carved out a niche for itself in the hot spring scene, where it’s been found living the high life in balmy waters. It’s a Gram-positive, strictly anaerobic, endospore-forming bacterium that doesn’t mind the heat, which suggests it knows how to handle a bit of thermal stress while it’s busy breaking down substances that would have most other organisms waving the white flag.

In the scientific community, C. sulfidigenes has gained a bit of a reputation as a multidrug-resistant organism, which means it’s not easily intimidated by antibiotics that would send other bacteria packing. This resilience, coupled with its metal resistance and chemical prowess, makes it an intriguing subject for researchers who are keen to understand how such a microscopic entity can stand up to environments and substances that are far from welcoming.

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Staphylococcus shinii with MALDI TOF MS

Staphylococcus shinii

Staphylococcus shinii is a newly proposed species of coagulase-negative staphylococci, isolated from fresh produce, particularly chives. It has been characterized for its antibiotic resistance genes and is of interest in the context of sulfur metabolism, which plays a significant role in the global sulfur cycle.
The bacterium was named in honor of Prof. Hyun-Kil Shin in recognition of his outstanding work in the area of food microbiology.
This species may possess antibiotic resistance genes and has been associated with nosocomial infections. It’s important to note that Staphylococcus shinii should not be confused with other staphylococci species, such as Staphylococcus epidermidis, or individuals like Shinji Okazaki, a Japanese footballer.

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Listeria aquatica identified with MALDI TOF MS

Listeria aquatica

Listeria aquatica was first described in 2014 together with L. floridensis, L. cornellensis, L. riparia and L. grandensis by M. Wiedmann et. al.
It is a Gram-positive, facultatively anaerobic, nonmotile, non-spore-forming rod-shaped bacterium. Interestingly, it is not pathogenic and belongs to the so called Listeria sensu lato species. The species was discovered in running water in Florida. The name “aquatica” comes from Latin, meaning “found in water, aquatic”.

One unique characteristic of Listeria aquatica is that it is the only member of the genus Listeria that can ferment maltose. It is also the only nonmotile Listeria that can ferment D-tagatose.
The type strain of Listeria aquatica is BEI NR-42633; DSM 26686; FSL S10-1188; LMG 28120.

Moreover, three sensu lato species (L. fleischmannii, L. floridensis, and L. aquatica) are unable to grow at low temperatures, one of the major characteristics of the genus Listeria. According to their phylogenetic and phenotypic characteristics a reclassification of L. fleischmannii, L. aquatica, and L. floridensis into the novelle genus Mesolisteria (referring to the mesophillic nature of species within this genus) is proposed.

It is crucial to monitor the presence of Listeria species in aquatic environments, including L. aquatica, as a potential indicator organism for the presence of L. monocytogenes and other pathogenic Listeria species.

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TODAY IN OUR LAB: Acetobacter orientalis

Acetobacter orientalis

Acetobacter orientalis is a species of bacteria that was first isolated from a canna flower, Canna hybrida. The name “orientalis” refers to the region where the strains were isolated.

This bacterium is a mesophilic species, meaning it grows best in moderate temperature conditions. It belongs to the genus Acetobacter, which is known for producing acid as a result of metabolic processes. Acetobacter is an obligatory aerobic, nitrogen-fixing bacteria and is characterized by the ability to convert ethanol to acetic acid in the presence of oxygen.
The type strain of this species is 21F-2; CIP 107379; DSM 15550; IFO 16606; JCM 11195; NBRC 16606; NRIC 481. The risk group for this bacterium is 1, indicating it’s not known to cause disease in healthy adult humans.

Acetobacter orientalis has several applications, particularly in the food and biotechnology industries:

Vinegar Production:

Strains of the genus Acetobacter, including A. orientalis, are used for vinegar fermentation due to their ability to oxidize ethanol to acetic acid and their high resistance to the resulting acetic acid.

Lactobionic Acid Production:

A. orientalis has been found to be involved in the production of lactobionic acid in Caucasian yogurt, also known as “Caspian Sea yogurt”, in Japan. Lactobionic acid is used in various applications, including as a component of the preservative solution used during organ transplantation and as an ingredient in the antibiotic erythromycin lactobionate.

Cellulose Degradation:

A strain of A. orientalis, XJC-C, has been found to have high-efficiency cellulase and ligninase activity, enabling it to degrade cellulose and lignin. This makes it useful for degrading various industrial and agricultural fertilizers containing cellulose and lignin, and for recycling byproducts and microbial fertilizers. This has practical value for waste treatment and high-efficiency utilization of microbial resources.

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TODAY IN OUR LAB: Cloacibacillus porcorum

Cloacibacillus porcorum

Isolated from a blood culture of a sepsis case in a swiss hospital.

Cloacibacillus porcorum is a species of bacteria that was first described by Looft et al. in 2013. It belongs to the genus Cloacibacillus. Cloacibacillus porcorum has been isolated from the intestinal tract of a pig from Ames in the United States. It is a Gram-negative, mesophilic (20°C-45°C), non-motile bacterium. It is known to degrade mucin, a type of protein produced by the epithelial tissues including those in the gastrointestinal tract.

The type strain of this species is CCUG 62631, CL-84, DSM 25858.
Cloacibacillus porcorum has been associated with human disease. It has been linked to rare cases of bacteremia. However, it’s important to note that the pathogenicity of this bacterium in humans is not fully understood and more research is needed to clarify its role in human disease.
Therefore, rapid identification of rare clinical germs is of great importance for appropriate treatment.

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TODAY IN OUR LAB: Zoogloea ramigera

Zoogloea ramigera

Taxonomy: Zoogloea ramigera was first described by Itzigsohn in 1868. The species is classified under the domain Bacteria, phylum Proteobacteria, class Betaproteobacteria, order Rhodocyclales, family Zoogloeaceae, and genus Zoogloea.

Characteristics: Zoogloea ramigera is a rod-shaped bacterium with actively mobile cells that possess a single polar flagellum. This bacterium forms characteristic cell aggregates surrounded by a gelatinous matrix, which gives it a “zoogloeal” appearance. It is a gram-negative bacterium, meaning it does not retain the crystal violet stain in the Gram staining process.

Habitat: Zoogloea ramigera is commonly found in organically enriched aqueous environments, such as activated sludges in sewage treatment plants. It can also be present in freshwater that contains organic matter. Additionally, Zoogloea ramigera has been isolated from forest soil, indicating its presence in natural environments as well.

Role in sewage treatment: Zoogloea ramigera has long been considered the typical activated sludge bacterium responsible for the formation of activated sludge flocs, which play a crucial role in wastewater treatment processes.

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