Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry for identification of Clostridium species isolated from Saudi Arabia

Abstract The aim of this study was to identify different Clostridium spp. isolated from currency notes from the Ha’il region of Saudi Arabia in September 2014 using MALDI–TOF-MS. Clostridium spp. were identified by Bruker MALDI–TOF-MS and compared with VITEK 2. The confirmation of the presence of different Clostridium spp. was performed by determining the sequence of the 16S ribosomal RNA gene. In this study, 144 Clostridium spp. were isolated. Among these specimens, MALDI–TOF-MS could identify 88.8% (128/144) of the isolates to the species level and 92.3% (133/144) to the genus level, whereas, VITEK 2 identified 77.7% of the (112/144) isolates. The correct identification of the 144 isolates was performed by sequence analysis of the 500 bp 16S rRNA gene. The most common Clostridium spp. identified were Clostridium perfringens (67.36%), Clostridium subterminale (14.58%), Clostridium sordellii (9%) and Clostridium sporogenes (9%). The results of this study demonstrate that MALDI–TOF-MS is a rapid, accurate and user friendly technique for the identification of Clostridium spp. Additionally, MALDI–TOF-MS has advantages over VITEK 2 in the identification of fastidious micro-organisms, such as Clostridium spp. Incorporating this technique into routine microbiology would lead to more successful and rapid identification of pathogenic and difficult to identify micro-organisms.

Acute myonecrosis in horse caused by Clostridium novyi type A

The objective of this study was to describe the first report involving a case of equine acute myonecrosis caused by C. novyi type A with an emphasis on clinical signs, the pathological and bacteriological analysis, and molecular identification of the microorganisms as the key of the definitive diagnosis.

First Korean Case of Robinsoniella peoriensis Bacteremia in a Patient with Aspiration Pneumonia

Robinsoniella peoriensis has recently been identified as a Gram-positive, spore-forming, anaerobic rod originally recovered from swine manure storage pits. To date, 6 cases of R. peoriensis infection have been reported, including 2 cases of bacteremia, 1 of abdominal fluid collection, and 3 of wound infection. In the present study, we report a 76-yr-old man with R. peoriensis bacteremia who developed aspiration pneumonia. Gram staining of a purified colony revealed Gram-positive, rod-shaped bacteria. Biochemical identification using API 20 A (bioMerieux, France) indicated presence of Clostridium spp. We performed both 500-bp and full-gene sequencing of 16S rRNA of the isolate. The sequence was analyzed with MicroSeq ID 16S rRNA Library v2.0 (Applied Biosystems, USA), GenBank Basic Local Alignment Search Tool (BLAST) (http://www.ncbi.nlm.nih.gov/genbank), and EzTaxon database v2.1 (http://www.eztaxon.org). The 500-bp 16S rRNA sequence of the blood culture isolate showed 99.16-99.79% similarity with R. peoriensis and the full-gene 16S rRNA sequence showed 98.87-99.50% similarity with R. peoriensis. The organism was confirmed as R. peoriensis by using all of the mentioned databases except for MicroSeq, which did not include the RNA sequence of this bacterium. This case suggests that identification of R. peoriensis might be challenging in clinical laboratories with no access to molecular methods, as certain commercial identification systems may not identify, or may misidentify, this organism. To the best of our knowledge, this is the first report of the isolation of R. peoriensis in Korea.

Clostrídios em condimentos utilizados em embutidos cárneos / Ciostridium of vegetable spices utilized in sausages

Em 35 amostras de diferentes condimentos vegetais, utilizados em embutidos cárneos, conforme metodologia recomendada por Holdeman et ai. (1977) e Jürgensen e Jürgensen (1982) foram isolados microrganismos do gênero Clostridium. Nas análises efetuadas comprovamos a presença de clostrídios em 65,7% do total de amostras analisadas. Foram isolados, pelas características fenotípicas identificadas, os seguintes microrganismos nas amostras correspondentes: Pimenta branca (Clostridium glycolicum, Clostridium butyricum, Clostridium perfringens), Louro (Clostridium perfringens); Canela (Clostridium pertríngens), Noz-moscada (Clostrídíum pertríngens, clostrídíum ramosum); Cominllo (Clostridium perene, Clostridium glycolicum, Cilostridium barati); Alho (Clostridium perene, Clostridium pertringens); Cebola (Clostridium glycolicum, Clostridium paraputrificum, Clostridium pertringens); Orégano (Clostridium acetobutylicum, Clostridium glycolicum, Clostridium ramos um); Coentro (Clostridium perfringens, Clostridium glycolicum); Pimenta preta (Clostridium feisineum, Clostridium sporogenes).

Using metllodology recommended by Holdeman et ai. (1977) and Jürgensen and Jürgensen (1982), micro-organisms of the genus Clostridium were isolated from thirty-five vegetal species utilized in sausage. Clostridium was seen in 65,7% of lhe samples analysed. The following micro-organisms (identied based on phenotypical characteristics) and corresponding vegetal samples were white pepper... (Clostridium glycolicum, Clostridium butyricum, Clostridium perfringens), laurel (Clostridium perfringens), Cinammon (Clostridium perfringens), Nutmeg (Ciostridium ramosum; Clostridium perfringens), Cummín (Clostridium perene, Clostridium glycolicum; Clostridium barati), Garlic (Clostridium perfringens, Clostridium perene), Onion (Clostridium perfringens, Clostridium glycolicum, Clostridium paraputrificum), Origanum (Clostridium ramosum, Clostridium glycolicum; Clostridium acetobutyricum), Coentro (Clostridium perfringens; Clostridium glycolicum), Black pepper (Clostridium felsineum and Clostridium sporogenes).