Sterility Test of Syringes As A Pharmaceutical Preparation That Obtained From Pasar Pramuka

Syringes are one of the pharmaceutical preparations that are in high demand. In healthcare institutions, syringes are used to aid in patient care and examination. Pharmaceutical preparations, such as various syringes, are widely available in the Pasar Pramuka. Syringes must be free from microbes and used syringes should not be reused. Microbiological sterility tests can be performed on a syringe to determine whether it is sterile or not. The purpose of the study is to test the sterility of syringes obtained from the Pasar Pramuka. A random sample of a syringe is being used in the study as an experimental method. The syringes were isolated and incubated for 14 days in Fluid Thioglycollate Medium (FTM) and Trypticase Soy Broth (TSB) at 30-35 oC and 20-25 oC, respectively, with frequent observations. If FTM and TSB media were turbid, then isolated into selective media based on their microbe as controls, namely Clostridium sporogenes ATCC 19404, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 25922, Escherichia coli ATCC 8739, Bacillus subtilis ATCC 6633, Salmonella typhi. The results showed that the B syringe was turbid on FTM media and did not contain pathogenic microbes after being identified on selective media, as in controls. The A and C syringes on TSB media were turbid, and after identification on selective media, Candida albicans and Aspergillus brasiliensis were found. In conclusion, the A, B, and C syringes are not steril.

Indole-3-Propionic Acid, a Functional Metabolite of Clostridium sporogenes, Promotes Muscle Tissue Development and Reduces Muscle Cell Inflammation

Clostridium sporogenes (C. sporogenes), as a potential probiotic, metabolizes tryptophan and produces an anti-inflammatory metabolite, indole-3-propionic acid (IPA). Herein, we studied the effects of C. sporogenes and its bioactive metabolite, IPA, on skeletal muscle development and chronic inflammation in mice. In the in vivo study, the muscle tissues and serum samples of mice with C. sporogenes supplementation were used to analyze the effects of C. sporogenes on muscle metabolism; the IPA content was determined by metabonomics and ELISA. In an in vitro study, C2C12 cells were exposed to lipopolysaccharide (LPS) alone or LPS + IPA to verify the effect of IPA on muscle cell inflammation by transcriptome, and the involved mechanism was revealed by different functional assays. We observed that C. sporogenes colonization significantly increased the body weight and muscle weight gain, as well as the myogenic regulatory factors’ (MRFs) expression. In addition, C. sporogenes significantly improved host IPA content and decreased pro-inflammatory cytokine levels in the muscle tissue of mice. Subsequently, we confirmed that IPA promoted C2C12 cells’ proliferation by activating MRF signaling. IPA also effectively protected against LPS-induced C2C12 cells inflammation by activating Pregnane X Receptor and restoring the inhibited miR-26a-2-3p expression. miR-26a-2-3p serves as a novel muscle inflammation regulatory factor that could directly bind to the 3′-UTR of IL-1β, a key initiator factor in inflammation. The results suggested that C. sporogenes with its functional metabolite IPA not only helps muscle growth development, but also protects against inflammation, partly by the IPA/ miR-26a-2-3p /IL-1β cascade.

Integration of Complete Plasmids Containing Bont Genes into Chromosomes of Clostridium parabotulinum, Clostridium sporogenes, and Clostridium argentinense

At least 40 toxin subtypes of botulinum neurotoxins (BoNTs), a heterogenous group of bacterial proteins, are produced by seven different clostridial species. A key factor that drives the diversity of neurotoxigenic clostridia is the association of bont gene clusters with various genomic locations including plasmids, phages and the chromosome. Analysis of Clostridium sporogenes BoNT/B1 strain CDC 1632, C. argentinense BoNT/G strain CDC 2741, and Clostridium parabotulinum BoNT/B1 strain DFPST0006 genomes revealed bont gene clusters within plasmid-like sequences within the chromosome or nested in large contigs, with no evidence of extrachromosomal elements. A nucleotide sequence (255,474 bp) identified in CDC 1632 shared 99.5% identity (88% coverage) with bont/B1-containing plasmid pNPD7 of C. sporogenes CDC 67071; CDC 2741 contig AYSO01000020 (1.1 MB) contained a ~140 kb region which shared 99.99% identity (100% coverage) with plasmid pRSJ17_1 of C. argentinense BoNT/G strain 89G; and DFPST0006 contig JACBDK0100002 (573 kb) contained a region that shared 100% identity (99%) coverage with the bont/B1-containing plasmid pCLD of C. parabotulinum Okra. This is the first report of full-length plasmid DNA-carrying complete neurotoxin gene clusters integrated in three distinct neurotoxigenic species: C. parabotulinum, C. sporogenes and C. argentinense.

Stability of Ready-to-Use and Laboratory-Prepared Culture Media

The culture media quality, which depends to a large extent on growth promotion properties, determines the reliability and accuracy of test results obtained by microbiological methods. The procedure for culture media preparation is quite labourconsuming and includes several stages: successive dissolution of components exactly as specified in the recipe (qualitative and quantitative composition), sterilisation, adjusting the pH of the medium, and testing of growth promotion properties—therefore it is important to demonstrate the stability of each particular culture medium.The aim of the study was to evaluate growth promotion properties of the culture media prepared in the laboratory from a dry mixture, and to assess their stability during long-term storage.Materials and methods: stability testing was performed for fluid thioglycollate medium (FTM) and soybean-casein digest broth (SCD) prepared in the laboratory. FTM growth promotion properties were tested using the following test microorganisms: Bacillus subtilis ATCC 6633, Clostridium sporogenes ATCC 19404, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538. SCD growth promotion properties were tested using: Aspergillus brasiliensis ATCC 16404, Candida albicans ATCC 10231, Bacillus subtilis ATCC 6633. The stability study was carried out for 6 months, assessing changes in appearance and growth promotion properties. The test media were stored at room temperature and in a refrigerator at 2–8 °C.Results: no growth of the anaerobic microorganism Clostridium sporogenes was observed in FTM after 3 months, regardless of storage conditions. Later on, there was no growth of the gram-positive microorganisms Bacillus subtilis and Staphylococcus aureus, which are more sensitive to the storage conditions than Pseudomonas aeruginosa. The growth promotion properties of the SCD prepared in the laboratory did not change during 6 months of storage.Conclusions: FTM prepared from a dry mixture remains stable and retains its growth promotion properties for no more than two months when stored in a refrigerator at 2–8 °C. SCD can remain stable for 6 months, both at room temperature and when stored in the refrigerator.

Comparative Evaluation of the Sterilization Effect of Six Kinds of Commercial Disinfectants against Four Pathogenic Bacteria

In this work, we systematically investigate the sterilization effect of six kinds of commonly used commercial disinfectants, including the DuPont Virkon disinfectant, peracetic acid disinfectant, sodium hypochlorite, bromogeramine disinfectant, water-soluble allicin, and absolute ethanol, against the Escherichia coli, Staphylococcus aureus, Monilia albican and Clostridium sporogenes. The inhibition zone was used to qualitatively determine the antibacterial effects of the six disinfectants, and then the minimum two-fold dilution method was used to quantitatively determine the minimum inhibitory concentration and minimum bactericidal concentration of the six disinfectants on the four pathogens. The result illustrated that the antibacterial effect of peracetic acid disinfectant is the best, and clostridium sporogenes is the most sensitive to it, followed by bromogermine disinfectant, which can inhibit the four pathogenic bacteria at the concentration recommended by the manufacturer. The antibacterial effect of DuPont Virkon disinfectant, sodium hypochlorite, water-soluble allicin and absolute ethanol is not as good as expected, and cannot inhibit the four kinds of pathogenic bacteria at the recommended concentration. In summary, the antibacterial effect of peracetic acid disinfectant is the strongest, followed by the bromogermine disinfectant, DuPont Virkon disinfectant, sodium hypochlorite and water-soluble allicin. The absolute ethanol exhibits the worst antibacterial properties.

Investigation of the Effectiveness of Disinfectants Used in Meat-Processing Facilities to Control Clostridium sporogenes and Clostridioides difficile Spores

Spore-forming bacteria are a major concern for the food industry as they cause both spoilage and food safety issues. Moreover, as they are more resistant than vegetative cells, their removal from the food processing environment may be difficult to achieve. This study investigated the efficacy of the ten most commonly used disinfectant agents (assigned 1–10), used at the recommended concentrations in the meat industry, for their ability to eliminate Clostridium sporogenes and Clostridioides difficile spores. Test-tube based suspension assays suggested that disinfectants 2 (10% v/v preparation of a mixture of hydrogen peroxide (10–30%), acetic acid (1–10%) and peracetic acid (1–10%)), 7 (4% w/v preparation of a mixture of peroxymonosulphate (30–50%), sulphamic acid (1–10%) and troclosene sodium (1–10%)) and 10 (2% v/v preparation of a mixture of glutaraldehyde (10–30%), benzalkonium chloride (1–10%)) were the most effective formulations. D-values for these ranged from 2.1 to 8.4 min at 20 °C for the target spores. Based on these findings, it is recommended that these disinfectants are used to control Clostridium spores in the meat plant environment.

Systematic dissection of a complex gut bacterial community

Efforts to model the gut microbiome have yielded important insights into the mechanisms of interspecies interactions, the impact of priority effects on ecosystem dynamics, and the role of diet and nutrient availability in determining community composition. However, the model communities studied to date have been defined or complex but not both, limiting their utility. Here, we construct a defined community of 104 bacterial strains composed of the most common taxa from the human gut microbiota. By propagating this community in growth media missing one amino acid at a time, we show that branched-chain amino acids have an outsize impact on community structure and identify a pathway in Clostridium sporogenes for generating ATP from arginine. We constructed and propagated the complete set of single-strain dropout communities, revealing a sparse network of strain-strain interactions including a novel interaction between C. sporogenes and Lactococcus lactis driven by metabolism. This work forms a foundation for studying strain-strain and strain-nutrient interactions in highly complex defined communities, and it provides a starting point for interrogating the rules of synthetic ecology at the 100+ strain scale.

Efficient Secretion of Murine IL-2 From an Attenuated Strain of Clostridium sporogenes, a Novel Delivery Vehicle for Cancer Immunotherapy

Despite a history dating back to the 1800s, using Clostridium bacteria to treat cancer has not advanced beyond the observation that they can colonise and partially destroy solid tumours. Progress has been hampered by their inability to eradicate the viable portion of tumours, and an instinctive anxiety around injecting patients with a bacterium whose close relatives cause tetanus and botulism. However, recent advances in techniques to genetically engineer Clostridium species gives cause to revisit this concept. This paper illustrates these developments through the attenuation of C. sporogenes to enhance its clinical safety, and through the expression and secretion of an immunotherapeutic. An 8.6 kb sequence, corresponding to a haemolysin operon, was deleted from the genome and replaced with a short non-coding sequence. The resultant phenotype of this strain, named C. sporogenes-NT, showed a reduction of haemolysis to levels similar to the probiotic strain, C. butyricum M588. Comparison to the parental strain showed no change in growth or sporulation. Following injection of tumour-bearing mice with purified spores of the attenuated strain, high levels of germination were detected in all tumours. Very low levels of spores and vegetative cells were detected in the spleen and lymph nodes. The new strain was transformed with four different murine IL-2-expressing plasmids, differentiated by promoter and signal peptide sequences. Biologically active mIL-2, recovered from the extracellular fraction of bacterial cultures, was shown to stimulate proliferation of T cells. With this investigation we propose a new, safer candidate for intratumoral delivery of cancer immunotherapeutics.