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Toxigenic members of Aspergillus flavus contaminate cereal grains, resulting in contamination by aflatoxin, a food safety hazard that causes hepatocellular carcinoma. This study identified probiotic strains as aflatoxin detoxifiers and investigated the changes to the grain amino acid concentrations during fermentation with probiotics in the presence of either A. flavus La 3228 (an aflatoxigenic strain) or A. flavus La 3279 (an atoxigenic strain). Generally, higher concentrations (p < 0.05) of amino acids were detected in the presence of toxigenic A. flavus La 3228 compared to the atoxigenic A. flavus La 3279. Compared to the control, 13/17 amino acids had elevated (p < 0.05) concentrations in the presence of the toxigenic A. flavus compared to the control, whereas in systems with the atoxigenic A. flavus 13/17 amino acids had similar (p > 0.05) concentrations to the control. There were interspecies and intraspecies differences in specific amino acid elevations or reductions among selected LAB and yeasts, respectively. Aflatoxins B1 and B2 were detoxified by Limosilactobacillus fermentum W310 (86% and 75%, respectively), Lactiplantibacillus plantarum M26 (62% and 63%, respectively), Candida tropicalis MY115 (60% and 77%, respectively), and Candida tropicalis YY25, (60% and 31%, respectively). Probiotics were useful detoxifiers; however, the extent of decontamination was species- and strain-dependent. Higher deviations in amino acid concentrations in the presence of toxigenic La 3228 compared to atoxigenic La 3279 suggests that the detoxifiers did not act by decreasing the metabolic activity of the toxigenic strain.

The goal is to study the features of the biological properties of Vibrio cholerae strains found in environmental objects (EO) on the territory of Rostov-on-Don from 1989 to 2018.Materials and methods. Bacteriological, molecular biological and statistical methods were used to study 73 strains of cholera vibrios O1, O139 and 1702 strains of nonO1/nonO139 serogroups isolated over a 30-year period.Results and discussion. It was found that in Rostovon-Don, 78.0 % of toxigenic strains were isolated from all isolated from environmental protection in Russia, among which 2 were genetically unchanged. The V. cholerae O1 classical strain and 72 V. cholerae El Tor strains were found, of which 70 % belonged to the Ogawa serovar, and 3 % of the nontoxigenic strains were susceptible to the classical phage. A “very weak direct relationship” was statistically revealed between the isolation of V. cholerae O1, O139 and V. cholerae nonO1/nonO139 strains, in which the dominant serogroups (O67, O76, O75, O53, O16) were identified, and 18 % of the strains showed atypical agglutinability. Revealing the biological characteristics of V. cholerae strains contributes to the optimization of monitoring studies.

The genus Corynebacterium includes species of biotechnological, medical and veterinary importance. An atypical C. ulcerans strain, W25, was recently isolated from a case of necrotizing lymphadenitis in a wild boar. In this study, we have analysed the genome sequence of this strain and compared the phenotypic and virulence properties with other corynebacterial pathogens. Phylogenomic analyses revealed that strain W25 belongs to a novel species along with PO100/5 and KL1196. The latter strains were isolated from a pig and a roe deer, respectively; hence, this species appears to be associated to animals. The isolate W25 is likely a non-toxigenic tox gene bearing strain and may have compromised abilities to adhere to pharyngeal and laryngeal epithelial cells due to potential loss of the gene functions in spaBC and spaDEF pilus gene clusters. A number of corynebacterial virulence genes are present including pld encoding phospholipase D. Therefore, this strain may be able to cause severe invasive infections in animals and zoonotic infections in humans.

The study was carried out to enumerate fungal contamination, and toxigenicity of Aspergillus flavus strains on cacao beans during drying and storage. As many as 3500 g cacao beans during drying and storage were purchased from smallholder farmers on the plantation areas at Karo Regency, Northern Sumatera. The percentage of the beans contaminated by fungi was conducted using direct plating. Fungal populations on soil and beans were determined using dilution followed by pour plated in dichloran 18% glycerol agar (DG18) and Aspergillus flavus and parasiticus agar (AFPA). The mycological evaluation was carried out based on morphological characteristics. Results showed eighteen genera of soil fungi were isolated at the cacao plantation; genera of Aspergillus sp., A. niger, A. flavus, and Penicillium citrinum were the most important contaminants. Six species of the fungi were associated with contamination on cocoa beans during drying i.e. Aspergillus sp., Candida tropicalis, Saccharomyces sp., A. niger, Penicillium spp., and Fusarium spp. Whereas three fungal species were associated during storage i.e. A. niger, A. flavus, and P. citrinum. The percentage of cacao contaminated during drying and storage was dominated (>40%) by Aspergillus sp. Fusarium sp. A. niger, A. flavus, and P. citrinum, respectively. Among 21 strains of A. flavus, 3 strains (15%) were isolated from soil, and 18 strains (85%) were isolated from beans during storage. Among toxigenic A. flavus, both strain scaf6 isolated from soil and strain cbaf5 isolated from beans during storage were the highest aflatoxin producers (30.0 ppb). Preventing soil contamination during harvesting, drying, and storage of cacao beans was a prerequisite to minimise fungal contamination.

We describe a case of interspecies transmission of toxigenic Clostridioides difficile involving a female and her dog, both with diarrhea without another diagnosis. Genomic analysis showed that isolates were grouped into MLST clade I, closely related to ribotype 020 and shared identical genotypes.

Aspergillus flavus is a phytopathogenic fungus able to produce aflatoxin B1 (AFB1), a carcinogenic mycotoxin that can contaminate several crops and food commodities. In A. flavus, two different kinds of strains can co-exist: toxigenic and non-toxigenic strains. Microbial-derived volatile organic compounds (mVOCs) emitted by toxigenic and non-toxigenic strains of A. flavus were analyzed by solid phase microextraction (SPME) coupled with gas chromatography–mass spectrometry (GC-MS) in a time-lapse experiment after inoculation. Among the 84 mVOCs emitted, 44 were previously listed in the scientific literature as specific to A. flavus, namely alcohols (2-methylbutan-1-ol, 3-methylbutan-1-ol, 2-methylpropan-1-ol), aldehydes (2-methylbutanal, 3-methylbutanal), hydrocarbons (toluene, styrene), furans (2,5-dimethylfuran), esters (ethyl 2-methylpropanoate, ethyl 2-methylbutyrate), and terpenes (epizonaren, trans-caryophyllene, valencene, α-copaene, β-himachalene, γ-cadinene, γ-muurolene, δ-cadinene). For the first time, other identified volatile compounds such as α-cadinol, cis-muurola-3,5-diene, α-isocomene, and β-selinene were identified as new mVOCs specific to the toxigenic A. flavus strain. Partial Least Square Analysis (PLSDA) showed a distinct pattern between mVOCs emitted by toxigenic and non-toxigenic A. flavus strains, mostly linked to the diversity of terpenes emitted by the toxigenic strains. In addition, the comparison between mVOCs of the toxigenic strain and its non-AFB1-producing mutant, coupled with a semi-quantification of the mVOCs, revealed a relationship between emitted terpenes (β-chamigrene, α-corocalene) and AFB1 production. This study provides evidence for the first time of mVOCs being linked to the toxigenic character of A. flavus strains, as well as terpenes being able to be correlated to the production of AFB1 due to the study of the mutant. This study could lead to the development of new techniques for the early detection and identification of toxigenic fungi.

Background The biological control mechanism of Aspergillus flavus (aflatoxigenic) strains in groundnuts with atoxigenic strains from the same species through competitive exclusion employed the use of endemic and well-adapted strains within the agro-ecological zones of Zimbabwe. The selected elite non-aflatoxigenic isolates of A. flavus native to Zimbabwe were evaluated for their capability to reduce aflatoxin contamination in groundnuts under laboratory conditions. Results Average reduction percentages in aflatoxin B concentration for the 2019 and 2020 set of experiments ranged from 91.6 ± 3.4 to 95.8 ± 3.1% and 90.29 ± 3.6% to 95.29 ± 4.1%, respectively. Levels of aflatoxin in the co-inoculation research experiments administered were significantly reduced in all the experimental units carried out. Treatment efficiencies of the tested isolates in this study at 4:1 and 2:1 ranged from 1.20 to 2.52 and from 1.02 to 1.21, respectively. The efficacy of the tested non-aflatoxigenic strains against the aflatoxigenic strain native to Zimbabwe (ZMW 0127) indicates that the non-aflatoxigenic isolates of A. flavus. have sound practical applications against vast communities of aflatoxin-producing fungi across all the agro-ecological zones in Zimbabwe. Conclusion The recognized non-aflatoxigenic isolates will be of an incentive as dynamic active ingredients in biocontrol formulations for the decrease in aflatoxins in groundnuts grown in Zimbabwe.

Aspergillus flavus is a phytopathogenic fungus able to produce aflatoxin B1 (AFB1), a carcinogenic mycotoxin that can contaminate several crops and food commodities. In A. flavus, two different kinds of strains can co-exist: toxigenic and non-toxigenic strains. Microbial-derived volatile organic compounds (mVOCs) emitted by toxigenic and non-toxigenic strains of A. flavus were analyzed by solid phase microextraction (SPME) coupled with gas chromatography–mass spectrometry (GC-MS) in a time-lapse experiment after inoculation. Among the 84 mVOCs emitted, 44 were previously listed in the scientific literature as specific to A. flavus, namely alcohols (2-methylbutan-1-ol, 3-methylbutan-1-ol, 2-methylpropan-1-ol), aldehydes (2-methylbutanal, 3-methylbutanal), hydrocarbons (toluene, styrene), furans (2,5-dimethylfuran), esters (ethyl 2-methylpropanoate, ethyl 2-methylbutyrate), and terpenes (epizonaren, trans-caryophyllene, valencene, α-copaene, β-himachalene, γ-cadinene, γ-muurolene, δ-cadinene). For the first time, other identified volatile compounds such as α-cadinol, cis-muurola-3,5-diene, α-isocomene, and β-selinene were identified as new mVOCs specific to the toxigenic A. flavus strain. Partial Least Square Analysis (PLSDA) showed a distinct pattern between mVOCs emitted by toxigenic and non-toxigenic A. flavus strains, mostly linked to the diversity of terpenes emitted by the toxigenic strains. In addition, the comparison between mVOCs of the toxigenic strain and its non-AFB1-producing mutant, coupled with a semi-quantification of the mVOCs, revealed a relationship between emitted terpenes (β-chamigrene, α-corocalene) and AFB1 production. This study provides evidence for the first time of mVOCs being linked to the toxigenic character of A. flavus strains, as well as terpenes being able to be correlated to the production of AFB1 due to the study of the mutant. This study could lead to the development of new techniques for the early detection and identification of toxigenic fungi.

Bu tez çalışması Bilimsel Araştırma Projeler Birimi tarafından 2015FBE046 nolu proje ile desteklenmiştir. Nesli tehlike altında olan deniz kaplumbağalarının korunmasında popülasyonunun istikrarlı ve artış gösterir halde olması önemlidir. Yuvalama alanlarının abiyotik faktörlere karşı korunması, biyotik faktörlerden ise yavru bireylerin korunarak yuvalardan yavru çıkış başarılarının arttırılması önem kazanmaktadır. Bu çalışmada yuva kumu, başarısız yumurta ve ergin bireylerden izole edilebilen mikrofungusların; bazı fiziksel parametreler ile embriyonik sürece olası etkileri araştırılmıştır. Bu amaç ile 2015 ve 2016 yuvalama sezonları boyunca Dalyan İztuzu Kumsalı’na yuvalama yapan İribaş deniz kaplumbağalarının (Caretta caretta) yuvalarından kuluçka öncesi kum örneği ile kuluçka sonrası kum örneği, ergin birey kloak sıvısı ve kuluçka süresi dolan yuvalardaki başarısız yumurtalar örneklenerek kolonize olan mikrofunguslar saptanmıştır. Toplam 51 yuvadan 885 başarısız yumurta örneği alınmış ve bu yumurtalar; 386 döllenmemiş, 327 erken dönem, 80 orta dönem ve 92 ileri dönem embriyo olarak dağılım göstermektedir. İlk önce temel mikrobiyolojik tanı ile cins düzeyinde belirlemeleri yapılan izolatlar daha sonra Kantitatif-PZR ile tür teşhisleri tamamlanmıştır. Yumurta, ergin kloak sıvısı, kuluçka öncesi kum ve kuluçka sonrası kum örneklerinden toplam 26 tür izole edilmiştir. Yumurta örneklerinden 16 tür, kuluçka öncesi kum ve kuluçka sonrası kum örneklerinden 11 ve ergin kloak sıvısı örneklerinden 16 tür izole edilmiştir. Mikrofungus izole edilen ergin kloak sıvısı ve yuva örneklerinde yavru çıkış başarısı düşük saptanmıştır. Kuluçkası tamamlanan yuvalarda mikrofungus artışı gözlenmiştir. İnsan aktivitesi yoğun bölgelerde yavru çıkış başarısı düşük saptanırken tersi durumda yavru çıkış başarısında artış hesaplanmıştır. Mikrofunguslar tür sayısı ile yavru çıkış başarısı arasında güçlü negatif korelasyon saptanmış ve yumurta örnekleri ile kum örnekleri arasında negatif yönlü ve düşük dereceli ilişki görülmüştür. The loggerhead sea turtle, Caretta caretta, is a threatened species. It is important to protect the nesting areas against abiotic and abiotic factors. In this study, sampled from nesting sand, failed eggs and adult cloaca liquids which can be colonised fungi, some physical parameters and probable effects on the embryonic process were investigated. For this aim, identification microfungi of colonized sampling from pre-laying nest sands, post-hatchling nest sands, adult cloaca liquid and failed eggs of loggerhead sea turtles nests at Dalyan Iztuzu Beach during the 2015-2016 nesting season. 885 failed eggs were taken from total of 51 nest sand these was 386 unfertilized, 327 early, 80 middle and 92 late embryos. The isolates were firstly identified by basic microbiological diagnosis and genus level, later species identification was completed by quantitatif PCR. Twenty six species were isolated from eggs, adult cloaca fluid, pre-nesting and post-hatchling nest sands. Sixteen species were isolated from adult cloaca fluid and egg samples, eleven species were isolated from nest sands. In cloaca fluid and nest samples isolated from microfungi, the success rate of hatchling was low. An increase in microfungi was observed in nests whose incubation was completed. In areas with high human activity, hatchling success was low, while an increase in hatchling success was calculated in the opposite case. A strong negative correlation was found between the number of microfungi species and hatchling success, and a negative and low-grade relationship was observed between egg and sand samples.

Azaspiracids (AZAs) are a group of lipophilic biotoxins responsible for the azaspiracid shellfish poisoning syndrome (AZP) in humans after consumption of contaminated shellfish. AZAs are produced by four representatives of the marine nanoplanktonic family Amphidomataceae (Dinophyceae), i.e. Azadinium spinosum, Az. poporum, Az. dexteroporum and Amphidoma languida. Among those species, Az. spinosum producing AZA-1, -2 and -33 (as known in 2017) and, to lesser extent, Az. poporum producing AZA-37, are known from the North Atlantic. These toxigenic species pose a major concern, especially for the coastal shellfish production in Ireland, and are thus frequently monitored along with AZA toxins by the regulatory authorities of the Irish government. A third North Atlantic AZA producer, Amphidoma languida, has been described based on an isolate obtained from Irish coastal waters, but the actual threat by this species and the respective AZA variants (AZA-38, -39) is unknown. In contrast to AZAs produced by Az. spinosum and Az. poporum, these AZA congeners are currently not regulated within the EU. The three AZA producers have been confirmed in the North Sea as well, but current knowledge on the biogeography of toxigenic Amphidomataceae relies on a limited number of observations and studies. The lack of data impedes an assessment of the actual risk of AZP in the North Sea and adjacent waters at present. However, shellfish farming in European coastal waters including the North Sea is of increasing importance for seafood supply, and enhanced production capacities are heavily advocated by the European Commission (EC). The goal of this thesis study was to increase knowledge about the current biogeography of toxigenic Amphidomataceae in the eastern North Atlantic, and to evaluate the risk potential of AZP in the area under the perspective of global change. Interpretations of the results should help to improve safety and sustainable use of coastal seafood production sites in the North Sea and adjacent areas. Major difficulties for reliable species detection and identification are the small cell size and inconspicuousness of nanoplanktonic Amphidomataceae, as well as the sympatric occurrence of toxigenic and non-toxigenic representatives. Multiple methods, i.e. light microscopy (LM) and scanning electron microscopy (SEM) for morphological inspection, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for AZA analysis, and quantitative polymerase chain reaction (qPCR) for DNA-based cell detection, were applied to respond to these challenges and to gain a broad spectrum of new insights into (toxigenic) Amphidomataceae. The isolation and characterization of (in total) 102 new Az. spinosum and Am. languida strains from the North Atlantic in 2016 and 2018 yielded increased knowledge on variation in AZA profiles and cell quotas of these toxigenic species. Samples from the North Sea provided 30 new Am. languida strains, all confirming previous morphological, phylogenetic and toxinological (i.e. AZA-38 and -39) records from the area. The 72 new Az. spinosum strains represented both Ribotype A in the North Sea and Irish Sea, but Ribotype B was only detected from the North Sea. For the first time, variability in the toxin profile of Ribotype A was confirmed, with different combinations of the three AZA variants (AZA-1 always present, combined with presence/absence of AZA-2 and/or -33), whereas the toxin profile of Ribotype B (AZA-11 and -51) was consistent in all strains. Multiple analyses over 18 months revealed that the AZA profile within all given strains remained stable. In contrast, AZA cell quotas were highly variable among and within Az. spinosum strains, and variability of single analogs was as high as 330-fold. These findings confirmed previous studies, but the reasons for the cell quota variability remain unclear. Five new amphidomatacean strains isolated from the 2018 field survey displayed the morphological characteristics of Az. spinosum, but exhibited significant DNA sequence differences (clustering closer to Az. obesum in phylogenetic trees) and no AZA production. The final taxonomic assignment remains undetermined, and the strains were thus designated as Az. cf. spinosum. The newly identified Az. cf. spinosum and the description of four new non-toxigenic Azadinium species (i.e. Az. galwayense, Az. perforatum, Az. perfusorium and Az. pseudozhuanum) highlighted in fact that amphidomatacean biodiversity is still underestimated and that AZA production is rather exceptional within this dinophyte family. Although qPCR assays for Az. spinosum and Az. poporum were already available prior to this study, the respective assay for quantification of toxigenic Amphidoma languida cells was developed and extensively evaluated in the course of this doctoral thesis project. A quick, cost-effective and high throughput application, coupled with high specificity and quantification limit down to 10 target gene copies per reaction, enables this sensitive assay to detect even single Am. languida cells per liter of seawater, and thus is a valuable tool for subsequent biogeographical studies. With respect to multiple newly discovered species and isolated amphidomatacean strains, specificity testing of the three alternative qPCR assays was of upmost importance to test for false-positive or falsenegative amplification and therefore to assure reliable detection and quantification in monitoring programs. None of the three assays showed false-positive signals, including for the new nontoxigenic Az. cf. spinosum, except for rDNA amplification from a new non-toxigenic Az. poporum isolate from the Danish coast. The most concerning result, however, was the significant amplification efficiency difference between Az. spinosum Ribotype A and B strains, revealing a degree of uncertainty for quantification from natural field samples by application of the current Az. spinosum assay because both ribotypes have been shown to co-occur in the Norwegian Sea and the North Sea. Although the current Az. spinosum and Az. poporum assays have not completely lost their validity for field applications, they should be redesigned for improved reliability. Multiple DNA sample sets, comprising more than 200 field samples from various expeditions between 2015 and 2019 to the eastern North Atlantic, were analyzed by qPCR for the presence and cell abundance of the three toxigenic amphidomatacean species. All three AZA-producers were found to be widely distributed in the area. In terms of positive geographical hits and cell densities (up to 8.3 x 104 cells L-1) Az. spinosum was the dominant toxigenic species in Irish coastal waters in summer 2018, underlining the threat for Irish shellfish production. Multiple hits and relatively high cell abundances of Az. spinosum were frequently found in the North Sea, as well. Amphidoma languida was also widely present and relatively abundant (2.3 x 104 cells L-1) around Ireland at that time, but highest cell density was found in the central North Sea, with an extraordinary abundance of ~ 1.2 x 105 cells L-1. This represents the highest ever recorded field abundance for this species and for North Atlantic Amphidomataceae in general. This finding, together with multiple further geographical records, indicated that Am. languida may be the dominant AZA producer in the North Sea. On this basis, incorporation of this species is recommended for both the national Irish- and official EU monitoring programs. Several amphidomatacean species have been found in Arctic and Subarctic waters before, and this finding was confirmed in the course of this study. Amphidoma languida was the only AZA producing species detected in the Arctic (> 75 °N) close to Spitzbergen in 2015, indicating that this species is able to cope with colder (around 5 °C) water temperatures. In contrast to Az. spinosum and Am. languida, Az. poporum was found in only a few locations and at low cell densities usually < 100 cells L-1, but with one extraordinary signal at Scapa Flow, Orkney Islands in June 2016, corresponding to ~ 3 x 103 cells L-1. This indicates an overall much lower potential contribution of this species to AZA contamination in recent years. Due to continuous sampling at several fixed North Sea stations, this thesis contains detailed qPCR data (in total 245 samples) on the seasonality of all three toxigenic species. The subsequent analysis revealed recurrent occurrence from July to October, consistent with observations at the Irish coastline (Marine Institute, Galway, Ireland), and indicating higher AZP risk in summer and fall. In addition, weekly sampling at the North Sea islands Helgoland and Sylt suggested relatively rapid population increases, demonstrating that sudden bloom events of toxigenic Amphidomataceae leading to rapid shellfish toxicity should be considered for respective monitoring frequency. First data on the vertical distribution of toxigenic Amphidomataceae presented here revealed no distinct distributional pattern in the water column, and hence pooling of water samples from various depths is an appropriate sampling method. Simultaneous on-board application of alternative technologies during an expedition in 2018 revealed a highly significant correlation between the results of light microscopy of plankton cells and qPCR assays for the detection and enumeration of toxigenic Amphidomataceae, and chemical analysis of AZA composition in the field. Detailed method-specific advantages and disadvantages are presented herein, but in particular the qPCR approach has proven to give solid results by combining high specificity with convenient detection limits. Laboratory experiments with North Atlantic strains representing all three toxigenic Amphidomataceae (including the first study on Am. languida) targeted temperature dependent growth and AZA production. Growth rates and AZA cell quota were inversely related: whereas higher temperatures led to higher growth rates, AZA content per cell decreased with increasing temperatures. Nevertheless, faster growth was shown to overcompensate for lower toxin cell quotas, leading to similar or even higher total AZA content per seawater volume (μg AZA L-1) at higher temperatures. This suggests a potentially increasing AZP risk under expected rising ocean temperatures. Highest AZA production was found in Az. spinosum Ribotype A (with a characteristic toxin profile of AZA-1, -2 and -33), highlighting a major role of this taxon determining AZP risk in the eastern North Atlantic. Except for Az. spinosum Ribotype B strain (containing AZA-11 and -51), all investigated strains showed lower extracellular than intracellular AZA levels. This suggests that AZA is predominantly retained intracellularly, and that screening for cells and intracellular AZAs is an appropriate monitoring method for AZP risk assessment. In conclusion, extensive research in this doctoral study, including development of a reliable qPCR assay for toxigenic Am. languida, with the description of new amphidomatacean species, strains, AZA variants, toxin profiles, adds considerably to the knowledge base on biogeography and variability within the Amphidomataceae. Combining data on AZA cell quota variability with the comprehensive data set on biogeography, seasonality and vertical distribution of the three toxigenic representatives in the North Sea has redefined our view of the role and importance of (toxigenic) Amphidomataceae and AZAs in the North Sea and adjacent areas. Thus, this doctoral thesis study provides a highly valuable baseline for official monitoring and future studies on toxigenic Amphidomataceae.

Corynebacterium ulcerans is an important zoonotic pathogen which is causing diphtheria-like disease in humans globally. In this study, the genomes of three recently isolated C. ulcerans strains, 4940, 2590 and BR-AD 2649, respectively from an asymptomatic carrier, a patient with pharyngitis and a canine host, were sequenced to investigate their virulence potential. A comparative analysis was performed including the published genome sequences of 16 other C. ulcerans isolates. C. ulcerans strains belong to two lineages; 13 strains are grouped together in lineage 1, and six strains comprise lineage 2. Consistent with the zoonotic nature of C. ulcerans infections, isolates from both the human and canine hosts clustered in both the lineages. Most of the strains possessed spaDEF and spaBC gene clusters along with the virulence genes cpp, pld, cwlH, nanH, rpfI, tspA and vsp1. The gene encoding Shiga-like toxin was only present in one strain, and 11 strains carried the tox gene encoding the diphtheria-like toxin. However, none of strains 4940, 2590 and BR-AD 2649 carried any toxin genes. These strains varied in the number of prophages in their genomes, which suggests that they play an important role in introducing diversity in C. ulcerans. The pan-genomic analyses revealed a variation in the number of membrane-associated and secreted proteins that may contribute to the variation in pathogenicity among different strains. Keywords: Corynebacterium ulcerans, diphtheria, nontoxigenic, sore throat, toxigenic, vaccine, virulence, zoonotic

Cholera is a severe diarrheal disease caused by Vibrio cholerae, a natural inhabitant of brackish water. Effective control of cholera outbreaks depends on prompt detection of the pathogen from clinical specimens and tracking its source in the environment. Although the epidemiology of cholera is well studied, rapid detection of V. cholerae remains a challenge, and data on its abundance in environmental sources are limited. Here, we describe a sensitive molecular quantification assay by qPCR, which can be used on-site in low-resource settings on water without the need for DNA extraction. This newly optimized method exhibited 100% specificity for total V. cholerae as well as V. cholerae O1 and allowed detection of as few as three target CFU per reaction. The limit of detection is as low as 5 × 103 CFU/L of water after concentrating biomass from the sample. The ability to perform qPCR on water samples without DNA extraction, portable features of the equipment, stability of the reagents at 4 °C and user-friendly online software facilitate fast quantitative analysis of V. cholerae. These characteristics make this assay extremely useful for field research in resource-poor settings and could support continuous monitoring in cholera-endemic areas.

Background: Clostridium difficile (CD) is a major cause of healthcare-associated infections and antibiotic-associated diarrhea in hospitalized patients worldwide. Carriers of toxigenic CD (tCD) have a higher risk of developing CD infections and can transmit CD to the environment and susceptible patients. However, little is known regarding the carriers and transmission of tCD in China. Methods: A multi-center cross-sectional study of tCD colonization (tCDC) was conducted from October 24 to 31, 2014, at 33 hospitals in Shanghai, China. Rectal swabs or stool samples were collected and tested, and the clinical and demographic status, epidemiological data, and blood parameters of 531 participants were recorded. The status of tCDC was defined by a positive result on the nucleic acid amplification test for the tcdA (toxin A), tcdB (toxin B), and cdtAB (toxin CDT) genes after positive bacterial culture. Results: The overall prevalence of CD colonization (CDC) was 19.02%, tCDC accounted for 92.08%, and A+B+CDT– was the dominant genotype (87.13%). The CD infection (CDI) prevalence was 1.51%. Potential tCDC-associated factors were admission to secondary grade hospitals, a body mass index

Cholera has been woven into human history through numerous pandemics, with the most recent ongoing since 1961. Global rates of cholera continue to decline, but outbreaks continue to pose diagnostic challenges for clinicians, which delays initiation of treatment and prolongs the disease course. Despite millions of infections and thousands of deaths worldwide each year, cholera remains rare in the United States, with the few cases each year usually being the result of pathogen acquisition while the patient traveled abroad. This article presents a unique case of cholera acquired in the United States, which emphasizes the necessary vigilance of symptom recognition, in the context of appropriate clinical investigation, in ensuring that the patient had a full recovery. Cholera in the United States is exceedingly rare, yet effective diagnosis with early initiation of treatment is known to reduce mortality and shorten disease course. While other more common diagnoses must definitely be excluded first, it is important for cholera to be kept on the differential for patients presenting with treatment refractory, watery diarrhea causing hypotension. This case of a patient with a recent travel history to Hawaii and infection with cholera underscores the importance of investigative medicine and clinical expertise in optimizing patient care, even when presented with rare illnesses.

Background Diphtheria has been reported as an outbreak in some regions in Indonesia, most especially in East Java Province. Resistance to penicillin, erythromycin, and other antibiotics, single or multiple, has been reported in several studies. This study aims to evaluate the first-line antibiotic susceptibility pattern of toxigenic Corynebacterium diphtheriae isolates. Methods This descriptive observational study was performed from August to November 2018. C. diphtheriae isolates were collected from diphtheria patients and carriers in East Java from 2012 to 2017 and kept at the Balai Besar Laboratorium Kesehatan Daerah Surabaya or the Public Health Laboratory of Surabaya. Sample selection was done by random cluster sampling. The sensitivity test by E-test®of the five antibiotics (penicillin, oxacillin, erythromycin, azithromycin, and clarithromycin) was done to determine the minimum inhibitory concentration (MIC). The Clinical and Laboratory Standards Institute M45A (2015) Corynebacterium spp. for penicillin and erythromycin was used as standard. Results From 114 targeted isolates, 108 were viable and toxigenic. The E-test was performed on the viable isolates. The majority of the hosts were male (58.3%), with median (range) age of 6.5 (1–14) years. Half of the samples were from the 1 to 5-year-old age group. The isolates were acquired much more from patients (78.7%) than carriers (21.3%) and from pharyngeal swab (74.1%). Most of these isolates were from Madura Island (47.2%) and the northern and eastern parts of the province (horseshoe area). Mitis isolates were the major variant (76.9%). The susceptibility pattern of C. diphtheriae to erythromycin was better than that to penicillin. The E-test result for penicillin was 68.52% susceptible, 31.48% intermediate, and 0% resistant (MIC range, 256 μg/L) was 85.2% susceptible, 12% intermediate, and 2.8% resistant The MIC range for oxacillin was 1 to 96 μg/L, while for both azithromycin and clarithromycin were 256 μg/L. Conclusion The susceptibility rate of C. diphtheriae to erythromycin is higher than that to penicillin. The regular update of antibiotic selection to the national guidelines is recommended. The MIC reference standard to azithromycin and clarithromycin is also needed.

Background: Clostridium difficile (CD) is a major cause of healthcare-associated infections and antibiotic-associated diarrhea in hospitalized patients worldwide. Carriers of toxigenic CD (tCD) have a higher risk of developing CD infections and can transmit CD to the environment and susceptible patients. However, little is known regarding the carriers and transmission of tCD in China. Methods: A multi-center cross-sectional study of tCD colonization (tCDC) was conducted from October 24 to 31, 2014, at 33 hospitals in Shanghai, China. Rectal swabs or stool samples were collected and tested, and the clinical and demographic status, epidemiological data, and blood parameters of 531 participants were recorded. The status of tCDC was defined by a positive result on the nucleic acid amplification test for the tcdA (toxin A), tcdB (toxin B), and cdtAB (toxin CDT) genes after positive bacterial culture. Results: The overall prevalence of CD colonization (CDC) was 19.02%, tCDC accounted for 92.08%, and A+B+CDT– was the dominant genotype (87.13%). The CD infection (CDI) prevalence was 1.51%. Potential tCDC-associated factors were admission to secondary grade hospitals, a body mass index

International audience; Epidemiology of diphtheria in the southwestern Indian Ocean is poorly documented. We analyzed 14 cases of infection with toxigenic Corynebacterium diphtheriae reported during 2007–2015 in Mayotte, a French department located in this region. Local control of diphtheria is needed to minimize the risk for importation of the bacterium into disease-free areas.

In September 2018, a child who had returned from Somalia to Germany presented with cutaneous diphtheria by toxigenic Corynebacterium diphtheriae biovar mitis. The child’s sibling had superinfected insect bites harbouring also toxigenic C. diphtheriae. Next generation sequencing (NGS) revealed the same strain in both patients suggesting very recent human-to-human transmission. Epidemiological and NGS data suggest that the two cutaneous diphtheria cases constitute the first outbreak by toxigenic C. diphtheriae in Germany since the 1980s.

Bovine mastitis has a negative impact on milk production and can pose risks to public health. The present study aimed to evaluate the quality of bovine milk from small farms in the Botucatu/SP region. Somatic cell counts (SCC), identification of pathogens involved in mastitis, and sensitivity antimicrobial profile of staphylococci isolated were performed. The presence of enterotoxin encoding genes in isolates of staphylococci obtained from milk was investigated. Milk samples from individual mammary quarters of cows were submitted to the California mastitis test (CMT) and SCC. Of the 239 dairy cows from 21 dairy herds evaluated (mean = 11.4 animals/property), two cows (0.8%) presented clinical mastitis and 86 (35.9%) subclinical mastitis. Bacterial culture was performed in 177 quarter milk samples. Staphylococci were identified in 55 (31.1%), corynebacteria in 45 (25.4%), streptococci in 25 (14.1%) and coliforms in four (2.3%) milk samples. Average SCC from culture-positive samples was 1598x103 cells/mL, in case of staphylococci was 1362x103 cells/ml, streptococci was 2857x103 cells/mL, corynebacteria was 976x103 cells/mL and in the cases of coliforms 1161x103 cells/mL were obtained. Staphylococci showed a high sensitivity (>95%) to cephalothin, cotrimoxazole, enrofloxacin, and gentamicin, with a 41.2% resistance to penicillin and 11.8% to oxacillin. Both coagulase positive (CPS) and negative staphylococci (CNS) carried genes encoding enterotoxins in 21.6% of the first group and 41.9% in the second. The sea gene was the most detected 45.8% (n=24) between them, followed by seb with 29.2% and sec with 25.0%. The sed gene was not identified. We highlight the potential risk to public health in the possibility of strains of Staphylococcus spp. enterotoxin-producing genes that can cause staphylococcal food poisoning. RESUMO: A mastite bovina impacta negativamente a produção leiteira e pode acarretar riscos à saúde pública. O presente estudo teve como objetivo a avaliação da qualidade do leite bovino proveniente de pequenas propriedades na região de Botucatu/SP. Foi realizada a contagem de células somáticas (CCS), identificação dos patógenos envolvidos nas mastites, e realizado o perfil de sensibilidade aos antimicrobianos dos estafilococos isolados. Pesquisou-se a presença de genes codificadores de enterotoxinas em isolados de estafilococos obtidos a partir do leite mastítico. Amostras de leite de quartos mamários individuais de vacas foram submetidas ao “California mastitis test” (CMT) e à CCS. Das 239 vacas em lactação provenientes de 21 rebanhos leiteiros avaliados (média = 11,4 animais/propriedade), dois (0,8%) animais apresentaram mastite clínica e, 86 (35,9%) mastite subclínica. 177 amostras de leite foram cultivadas em ágar sangue bovino 5% e ágar MacConckey e obteve-se 55 (31,1%) Staphylococcus spp., 25 (14,1%) Streptococcus spp., 45 (25,4%) Corynebacterium spp. e quatro (2,3%) coliformes. A média da CCS das amostras procedentes de todos os quartos mamários infectados avaliados foi de 1598x103 células/mL, enquanto que nos casos que foram isolados Staphylococcus spp. foi de 1362x103 células/mL, Streptococcus spp. 2857x103 células/mL, Corynebacterium spp. de 976x103 células/mL e nos casos de coliformes 1161x103 células/mL. Os estafilococos revelaram grande sensibilidade (>95%) à cefalotina, cotrimoxazol, enrofloxacina e gentamicina, com resistência de 41,2% à penicilina e 11,8% à oxacilina. Tanto estafilococos coagulase positivos (ECP) como negativos (ECN) revelaram genes codificadores de enterotoxinas em 21,6% do primeiro grupo e 41,9% no segundo. O gene sea foi o mais detectado 45,8% (n=24), seguido pelo seb com 29,2% e sec com 25,0%. O gene codificador da sed não foi identificado. Frente aos resultados, destaca-se o risco potencial à saúde pública pela possibilidade de veiculação de linhagens de Staphylococcus spp. carreadores de genes produtores de enterotoxinas, podendo ocasionar toxi-infecções alimentares.

In early 2017, a United Kingdom (UK)-born person in their 20s presented with a skin ulcer on the foot 3 weeks after returning from Ghana. The patient had last received a diphtheria-containing vaccine in 2013, completing the recommended course. MALDI-TOF of a cutaneous swab identified Corynebacterium diphtheriae. Real-time PCR ascertained the species and presence of the diphtheria toxin gene. An Elek test confirmed toxigenicity. The isolate was macrolide sensitive and penicillin resistant. The local Public Health England (PHE) Health Protection Team obtained the patient’s clinical history and traced contacts to inform appropriate public health action. One close contact (in their early 80s with uncertain immunisation status who had not recently travelled) had a positive throat swab for toxigenic C. diphtheriae and reported a history of mild coryzal symptoms. Multilocus sequence typing revealed that strains from the index case and contact had Sequence Type 493*. Diphtheria is extremely rare in the UK due to high vaccine coverage and this is the first documented transmission in 30 years. Clinicians and laboratory staff should remain highly suspicious of lesions in overseas travellers, even when patients are fully vaccinated. Older individuals who might not have completed a full immunisation course may have higher diphtheria susceptibility.

Objective There is a lack of study in Corynebacterium diphtheriae isolates in Malaysia. The alarming surge of cases in year 2016 lead us to evaluate the local clinical C. diphtheriae strains in Malaysia. We conducted single nucleotide polymorphism phylogenetic analysis on the core and pan-genome as well as toxin and diphtheria toxin repressor (DtxR) genes of Malaysian C. diphtheriae isolates from the year 1986–2016. Results The comparison between core and pan-genomic comparison showed variation in the distribution of C. diphtheriae. The local isolates portrayed a heterogenous trait and a close relationship between Malaysia’s and Belarus’s, Africa’s and India’s strains were observed. A toxigenic C. diphtheriae clone was noted to be circulating in the Malaysian population for nearly 30 years and from our study, the non-toxigenic and toxigenic C. diphtheriae strains can be differentiated significantly into two large clusters, A and B respectively. Analysis against vaccine strain, PW8 portrayed that the amino acid composition of toxin and DtxR in Malaysia’s local strains are well-conserved and there was no functional defect noted. Hence, the change in efficacy of the currently used toxoid vaccine is unlikely to occur.

Introduction: Toxigenic Clostridium difficile is the major cause of antibiotic-associated diarrhea, colitis, and pseudomembranous colitis. The pathogenicity of C. difficile is related to toxins A&B. Children with cancer are at risk of developing C. difficile infection (CDI) due to increased exposure to antibiotics, immunosuppression, and longer hospital stays. Recently, due to higher sensitivity and specificity of nucleic acid amplification test (NAATs) compared to toxin enzyme immunoassays (EIAs), many laboratories are transitioning to NAATs for detection of CDI. We aimed to use a multiplex PCR to detect the C. difficile toxin genes tcdA, tcdB and tpi in stool of cancerous children. We also aimed to show the effects of chemotherapy regimens on the prevalence of C. difficile in these children. Methods: 105 fecal samples were collected from cancerous children who were hospitalized and undergoing chemotherapy. The presence of tcdA, tcdB, and tpi genes were tested. Results: C. difficile was identified in 17.14% of children and the detection rate of A-B+ strains was higher than A+B+ and A+B- strains. C. difficile was found in 17.8% of children who received single antibiotic (5/28 cases virulence genes were detected in 4 cases) and in 41.4% of patients who received more than one antibiotics (12/29 cases virulence genes were detected in 8 cases﴿. Conclusion: Multiplex PCR is a powerful technique for preliminary screening and rapid detection of C. difficile and its virulence genes in the stool of cancerous children. The prevalence of C. difficile in cases receiving several antibiotics was higher than those receiving single antibiotics.

Background Diphtheria remains a major public health concern with multiple recent outbreaks around the world. Moreover, invasive non-toxigenic strains have emerged globally causing severe infections. A diphtheria epidemic in the former Soviet Union in the 1990s resulted in ~5000 deaths. In this study, we analysed the genome sequences of a collection of 93 C. diphtheriae strains collected during and after this outbreak (1996 – 2014) in a former Soviet State, Belarus to understand the evolutionary dynamics and virulence capacities of these strains. Results C. diphtheriae strains from Belarus belong to ten sequence types (STs). Two major clones, non-toxigenic ST5 and toxigenic ST8, encompassed 76% of the isolates that are associated with sore throat and diphtheria in patients, respectively. Core genomic diversity is limited within outbreak-associated ST8 with relatively higher mutation rates (8.9 × 10−7 substitutions per strain per year) than ST5 (5.6 × 10−7 substitutions per strain per year) where most of the diversity was introduced by recombination. A variation in the virulence gene repertoire including the presence of tox gene is likely responsible for pathogenic differences between different strains. However, strains with similar virulence potential can cause disease in some individuals and remain asymptomatic in others. Eight synonymous single nucleotide polymorphisms were observed between the tox genes of the vaccine strain PW8 and other toxigenic strains of ST8, ST25, ST28, ST41 and non-toxigenic tox gene-bearing (NTTB) ST40 strains. A single nucleotide deletion at position 52 in the tox gene resulted in the frameshift in ST40 isolates, converting them into NTTB strains. Conclusions Non-toxigenic C. diphtheriae ST5 and toxigenic ST8 strains have been endemic in Belarus both during and after the epidemic in 1990s. A high vaccine coverage has effectively controlled diphtheria in Belarus; however, non-toxigenic strains continue to circulate in the population. Recombination is an important evolutionary force in shaping the genomic diversity in C. diphtheriae. However, the relative role of recombination and mutations in diversification varies between different clones. Electronic supplementary material The online version of this article (10.1186/s12864-017-4276-3) contains supplementary material, which is available to authorized users.

Aim: To examine powdered infant formula (PIF) and follow-up formula (FUF) for the presence of toxigenic Clostridiumdifficile.Materials and Methods: A total of 100 random samples of PIF and FUF, 50 samples each, from various pharmacies andsupermarkets located in Assiut city were collected during 2008-2010.Results: Our results show that 16 out of 100 (16%) examined samples of PIF and FUF were contaminated with C. difficile; 4(8%) and 12 (24%) of the examined PIF and FUF samples tested positive for C. difficile, respectively. Only two (16.67%)isolates of C. difficile from the examined FUF were toxigenic, while the isolates from the PIF samples were not toxigenic.Conclusions:The presence of C. difficile in PIF and FUF samples suggests that there is a high potential for the transmission ofC. difficile through these products. Thus, proper preparation and handling of these products is required to reduce the risk of theillnesses arising due to C. difficile.

Organisms, including Vibrio cholerae, can be transferred between harbors in the ballast water of ships. Zones in the Caribbean region where distance from shore and water depth meet International Maritime Organization guidelines for ballast water exchange are extremely limited. Use of ballast water treatment systems could mitigate the risk for organism transfer.

Severe necrotizing fasciitis was diagnosed in a 53-year-old man in Germany in 2012. Toxigenic Corynebacterium ulcerans was grown from a wound swab sample. One of the patient´s 2 dogs was found to harbor a toxigenic C. ulcerans strain. Results of next generation sequencing of both isolates supported recent zoonotic transmission of this bacterial pathogen.

An epidemic of cholera infections was documented in Haiti for the first time in more than 100 years during October 2010. Cases have continued to occur, raising the question of whether the microorganism has established environmental reservoirs in Haiti. We monitored 14 environmental sites near the towns of Gressier and Leogane during April 2012–March 2013. Toxigenic Vibrio cholerae O1 El Tor biotype strains were isolated from 3 (1.7%) of 179 water samples; nontoxigenic O1 V. cholerae was isolated from an additional 3 samples. All samples containing V. cholerae O1 also contained non-O1 V. cholerae. V. cholerae O1 was isolated only when water temperatures were ≥31°C. Our data substantiate the presence of toxigenic V. cholerae O1 in the aquatic environment in Haiti. These isolations may reflect establishment of long-term environmental reservoirs in Haiti, which may complicate eradication of cholera from this coastal country. Download MP3 Length: 1:33

Corynebacterium ulcerans may cause diphtheria in humans and caseous lymphadenitis in animals. We isolated nontoxigenic tox-bearing C. ulcerans from 13 game animals in Germany. Our results indicate a role for game animals as reservoirs for zoonotic C. ulcerans.

We resolved the relationships between 2 pandemic clones of Vibrio cholerae. Using 26 housekeeping genes, we showed that the US Gulf clone, the Australian clone, and 3 El Tor strains isolated before the seventh pandemic were related to the seventh pandemic clone. The sixth pandemic clone was well separated from them.

Endemic oregano's, Origanum hypericifolium O. Schwartz and P.H. Davis, essential oil was extracted to exert its biological activity in vitro. Fifteen components in its extracts performed by hydro distillation. The major components in the fruit and flower volatiles of O. hypericifolium were p-cymene (34.33 g/100 g oil), carvacrol (21.76 g/100 g oil), thymol (19.54 g/100 g) and -terpinene (13.91 g/100 g oil). The antifungal activity of O. hypericifoliums oil was evaluated against 14 fungi isolated from hazelnut and walnut. Nuts are capable of harboring toxigenic fungi and the threat of mycotoxin contamination on them exists. Essential oil of O. hypericifolium was found to be active both in contact and headspace assays in vitro producing hyphal growth inhibition. In the contact assay, P. frequentans was found to be the least sensitive species. The more sensitive species were P. castellonense, P. verrucosum. var. cyclopium, C. globosum, and A. kiliense. Their growth was completely (100%) inhibited at days 3 and 6. In the volatile assay, all the mycelial growth of all tested fungi was completely inhibited at day 3. The volatile activity was found to be highly efficient than that of contact activity assay. This could be because of the aromatic contents of Origanum, such as monoterpenes, carvacrol, thymol, and p-cymene. © 2012 Copyright Taylor and Francis Group, LLC.

To the Editor: In the March 2014 issue of Emerging Infectious Diseases, Alam et al. reported a survey of water sources in Haiti conducted to isolate Vibrio cholerae (1). Each month from April 2012 through March 2013, they sampled 15 sites at 3 rivers and 1 estuary in West Department. From 179 water samples and 144 aquatic animals and plants, they obtained 7 V. cholerae O1 isolates, including 3 ctx-positive toxigenic strains. Unfortunately, the results for all 7 V. cholerae O1 isolates were aggregated, and no details were provided about the exact time and location of collection of samples corresponding to the 3 ctx-positive strains. The authors posed the question of whether V. cholerae O1 has become established in environmental reservoirs in Haiti, subsequently warning that “as long as the causative microorganism is present in the environment, eradication of the disease will not be possible.” However, after challenging their results with more accurate epidemiologic data, we found that these 3 ctx-positive toxigenic strains could more likely have been present in the sampled rivers as a result of recent fecal contamination (Figure). Indeed, many cholera cases were reported in the corresponding communal sections (i.e., the smallest Haitian administrative unit, average 25 km2) when the samples containing the 7 V. cholerae O1 isolates were collected. In this context of an ongoing cholera epidemic associated with persisting rainfall (Figure), generalized open-air defecation inevitably leads to contamination of water sources. It is therefore impossible to determine whether V. cholerae–positive rivers constitute perennial reservoirs of the bacteria or whether they act only as transient vectors of the pathogens. Figure Weekly cholera incidence in communal sections of Haiti in which water samples were positive for Vibrio cholerae O1; accumulated precipitation in the studied area by week during April 2012–March 2013; and number of environmental sites from which ... The recent dramatic decrease in cholera transmission may provide a good opportunity to address this issue (2). We thus encourage Alam et al. to continue the search for ctx-positive toxigenic V. cholerae O1 strains in surface waters, especially during cholera-free periods.

Vibrio cholerae is the etiological agent of cholera that has emerged as an endemic disease in different regions of the world in recent years. Traditional microbial culture and microscopy methods are considered to be the best standard for diagnosing V. cholerae infection. These methods, however, delay any available confirmatory answer by days. Molecular methods have the potential to provide sensitive, accurate, and rapid analysis of V. cholerae infection. We have developed a multiplex PCR assay to detect virulence and toxigenic-associated (VTA) genes (ctxA, tcpA, and ompW). To evaluate PCR specificity, additional bacteria from the enterobacteriaceae family (Salmonella typhi, Shigella dysantry, and entrotoxigenic E. coli) and Aeromonas hidrophyla were examined in this study. Specificity tests were evaluated using the genome dilution method. Importantly, the results show that our PCR specificity method represents the best tool for the rapid detection of VTA genes because of its simplicity, cost effectiveness, and accuracy. This multiplex PCR method can be used for examining the existence of VTA genes in patient samples, and therefore will distinguish V. cholerae from other vibrios and bacteria. This method is able to detect 10-100 colony forming units (CFUs) of V.Cholerae and 8.5-85 picograms (pg) of genomic DNA. The multiplex PCR method is also more specific and sensitive than other methods, validating it as an appropriate and sensitive tool for detecting the presence of toxigenic and pathogenic V. cholerae.

Diatoms responsible for harmful events in other geographic areas, found in the coastal waters of the San Matías Gulf (Río Negro Province, Argentina), were studied. Materials were collected in two sampling stations related to natural banks and cultures of bivalves: Las Grutas and Piedras Coloradas. Two potentially toxigenic taxa and 18 harmful non toxigenic taxa were found. Despite that some of these taxa were occasionally abundant in the plankton of the area, none of them caused evident discolorations or detectable harmful effects during the study period. Este trabajo tuvo como objetivo estudiar las diatomeas presentes en las aguas costeras del área norte del Golfo San Matías (Provincia de Río Negro, Argentina) que han sido citadas como productoras de eventos nocivos en otras áreas geográficas. El material fue recolectado en dos estaciones de muestreo: Las Grutas y Piedras Coloradas, relacionadas con bancos naturales y cultivo de bivalvos respectivamente. Dos taxa potencialmente toxígenos y 18 taxa nocivos no toxígenos fueron hallados. A pesar de que algunos de ellos fueron ocasionalmente abundantes en el plancton del área, ninguno produjo discoloraciones evidentes ni se detectó que hubieran causado efectos nocivos durante el período de estudio. Facultad de Ciencias Naturales y Museo

Diatoms responsible for harmful events in other geographic areas, found in the coastal waters of the San Matías Gulf (Río Negro Province, Argentina), were studied. Materials were collected in two sampling stations related to natural banks and cultures of bivalves: Las Grutas and Piedras Coloradas. Two potentially toxigenic taxa and 18 harmful non toxigenic taxa were found. Despite that some of these taxa were occasionally abundant in the plankton of the area, none of them caused evident discolorations or detectable harmful effects during the study period., Este trabajo tuvo como objetivo estudiar las diatomeas presentes en las aguas costeras del área norte del Golfo San Matías (Provincia de Río Negro, Argentina) que han sido citadas como productoras de eventos nocivos en otras áreas geográficas. El material fue recolectado en dos estaciones de muestreo: Las Grutas y Piedras Coloradas, relacionadas con bancos naturales y cultivo de bivalvos respectivamente. Dos taxa potencialmente toxígenos y 18 taxa nocivos no toxígenos fueron hallados. A pesar de que algunos de ellos fueron ocasionalmente abundantes en el plancton del área, ninguno produjo discoloraciones evidentes ni se detectó que hubieran causado efectos nocivos durante el período de estudio., Facultad de Ciencias Naturales y Museo

Las enfermedades diarreicas constituyen una causa frecuente de mortalidad y morbilidad en niños. Su etiología no siempre se logra identificar sugiriendo la presencia de otros enteropatógenos desconocidos. El aislamiento de Acinetobacter spp. en heces de niños con diarrea aguda no asociada a enteropatógenos clásicos, motivó su investigación a fin de detectar un posible efecto tóxico sobre células HT-29. El estudio estuvo constituido por 150 niños en edades entre 0 a 5 años; 120 fueron asistidos en el Hospital Materno Infantil del Este, Caracas, Venezuela, por presentar síndrome gastrointestinal y 30 eran niños sanos que acudían a dicho centro por control de rutina. En el 25% de los pacientes sintomáticos se diagnosticaron parásitos y bacterias de identificación rutinaria. De cuatro pacientes sintomáticos se aislaron tres cepas de Acinetobacter baumannii y dos de Acinetobacter calcoaceticus, estas cepas se cultivaron durante 24 y 48 horas a 35°C en caldo infusión cerebro corazón de donde se obtuvieron por centrifugación y filtración los sobrenadantes cuya actividad se ensayó en células HT-29. Los sobrenadantes de las tres cepas de A. baumannii presentaron profundas alteraciones en la monocapa celular, manifestada por el desprendimiento de la misma, disgregación, redondeamiento y expansión de las células. La toxicidad del sobrenadante de la cepa 016 superó la de los controles positivos. Este efecto toxigénico de A. baumannii, podría representar un mecanismo de patogenicidad cuya definición requiere de posteriores estudios para determinar un posible papel en la patogenia de este bacilo. Diarrheal diseases with diarrhea are the most frequent cause of morbidity and mortality in children; however the causative agent cannot be identified always, which suggests the presence of unknown enteropathogens inducing diarrhea. The isolation of Acinetobacter sp. from feces of children with acute diarrhea, unrelated to known enteropathogens motivated this investigation to detect a possible enterotoxigenic effect on HT-29 cells. The study population comprised 150 children with an age range from 0 to 5 years old; 120 were assisted in the “Hospital Materno Infantil del Este with gastrointestinal syndrome and 30 healthy controls who went to the center for routine analysis. In 25% of symptomatic patients were diagnosed parasites and bacteria, identified routinely. From four symptomatic patients were isolated three Acinetobacter baumannii strains and two A. calcoaceticus strains. The strains were cultured in brain-heart infusion for 24 and 48 hrs, at 35°C, and the supernatants were obtained by centrifugation and filtration and their activity tested on HT-29 cell monolayers. The supernatants of the three strains of A. baumannii induced alterations of the cell monolayer, showed by detachments of cell monolayers, cell segregation, cell rounding and swelling. These effects were more intense with the 48 h culture exoproducts of the 016 strain, which were higher than the positive control. This toxigenic effect of A. baumannii, could represent a pathogenic mechanism whose definition requires more studies to determine the possible role in the pathogenicity of this bacillus.

Undersøkelsen ble utført for Vestfold interkommunale vannverk for å fremskaffe en løpende bedømmelse av råvannskvaliteten i Akersvatnet. Månedlige situasjonsbeskrivelser ble utarbeidet. Sommeren 1997 ble den varmeste på Østlandet på over hundre år. Masseutvikling av planktonalger, med Ceratium hirundinella og blågrønnalger i dominans, medførte bl.a. en kritisk situasjon for fiskebestanden i perioden juli-august. Praktiske tiltak ble iverksatt, fiskedød inntraff ikke. Vannkvaliteten var i perioden med masseutvikling av alger uegnet som råvann til Akersvannverket. Cyanotoksiner ble påvist i blågrønalgene som utviklet seg. Aphanizomenon flos-aquae fikk oppmerksomhet som mulig ny toksinproduserende art i Akersvatnet. Vestfold interkommunale vannverk (VIV)

Isolates of Clostridium difficile from 159 hospitalized Danish patients (2005) were analysed by a new 5-plex PCR method targeting the toxin genes tcdA, tcdB, cdtA and cdtB, and 16S rDNA as an internal positive control. Additionally, the toxin-regulating gene tcdC was partially sequenced by a new sequencing-based method that revealed genetic changes that may render the gene product inactive. Finally tcdA was analysed using a previously published method for the detection of internal deletions. The 5-plex PCR revealed four different toxin gene profiles: 36 tcdA+, tcdB+, cdtA+ / cdtB+; one tcdA+, tcdB–, cdtA+ / cdtB+; 98 tcdA+, tcdB+, cdtA– / cdtB–; and 24 non-toxigenic tcdA–, tcdB–, cdtA– / cdtB–. Deletion studies revealed that 26 strains contained a c. 700-bp deletion in tcdA, and 39 strains contained at least one possible inactivation feature in tcdC. The prevalence of the binary toxin genes was 23%. All strains with the tcdA+, tcdB+, cdtA+ / cdtB+ profile were investigated by PCR ribotyping, and this revealed eight different ribotypes, none of which were 027. The 5-plex PCR method offers a one-step, rapid and specific screening method for C. difficile toxin genes. This toxin gene profiling, together with deletion studies in tcdA and tcdC, may allow an evaluation of the pathogenic potential of C. difficile.