scholarly journals Spatial, Temporal, and Matrix Variability of Clostridium botulinum Type E Toxin Gene Distribution at Great Lakes Beaches

2015 ◽  
Vol 81 (13) ◽  
pp. 4306-4315 ◽  
Author(s):  
Rasanthi U. Wijesinghe ◽  
Ryan J. Oster ◽  
Sheridan K. Haack ◽  
Lisa R. Fogarty ◽  
Taaja R. Tucker ◽  
...  
Keyword(s):  
Great Lakes ◽  
Clostridium Botulinum ◽  
Lake Michigan ◽  
Dry Weight ◽  
Toxin Gene ◽  
Average Water Temperature ◽  
Content Type ◽  
Gene Copies ◽  
Clostridium Botulinum Type E ◽  
City State

ABSTRACTClostridium botulinumtype E toxin is responsible for extensive mortality of birds and fish in the Great Lakes. TheC. botulinum bontEgene that produces the type E toxin was amplified with quantitative PCR from 150 sloughed algal samples (primarilyCladophoraspecies) collected during summer 2012 from 10 Great Lakes beaches in five states; concurrently, 74 sediment and 37 water samples from four sites were also analyzed. ThebontEgene concentration in algae was significantly higher than in water and sediment (P< 0.05), suggesting that algal mats provide a better microenvironment forC. botulinum. ThebontEgene was detected most frequently in algae at Jeorse Park and Portage Lake Front beaches (Lake Michigan) and Bay City State Recreation Area beach on Saginaw Bay (Lake Huron), where 77, 100, and 83% of these algal samples contained thebontEgene, respectively. The highest concentration ofbontEwas detected at Bay City (1.98 × 105gene copies/ml of algae or 5.21 × 106g [dry weight]). This study revealed that thebontEgene is abundant in the Great Lakes but that it has spatial, temporal, and matrix variability. Further, embayed beaches, low wave height, low wind velocity, and greater average water temperature enhance thebontEoccurrence.

CLOSTRIDIUM BOTULINUM FOOD POISONING1,2

1965 ◽  
Vol 28 (3) ◽  
pp. 86-91 ◽  
Author(s):  
E. M. Foster ◽  
Janet S. Deffner ◽  
Thomas L. Bott ◽  
Elizabeth McCoy
Keyword(s):  
Great Lakes ◽  
Clostridium Botulinum ◽  
Intestinal Tract ◽  
Lake Michigan ◽  
The United States ◽  
Main Body ◽  
Smoked Fish ◽  
Food Borne ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E

Summary The outbreaks of botulism in the United States during 1963 stimulated renewed interest in this food-borne disease, primarily because commercially prepared foods were involved. Three of the outbreaks were caused by Clostridium botulinum type E in fishery products. Two of these resulted from the consumption of smoked fish from the Great Lakes. A survey has been started to see if C. botulinum type E is common on fish from the Great Lakes. Toxin neutralization tests have shown the organism to be present in cultures from nine of ten locations sampled in Lake Michigan. The organism was found more frequently in the intestinal tract than on gills, livers or the external surfaces of the fish. Over 75% of the cultures prepared from the intestines of fish caught in one large bay of Lake Michigan proved to contain type E toxin. The incidence of the organism in fish from the main body of the lake has been much lower than this.

Effect of Salt Level and Nitrite on Toxin Production by Clostridium botulinum Type E Spores in Smoked Great Lakes Whitefish1

1987 ◽  
Vol 50 (3) ◽  
pp. 212-217 ◽  
Author(s):  
S. L. CUPPETT ◽  
J. I. GRAY ◽  
J. J. PESTKA ◽  
A. M. BOOREN ◽  
J. F. PRICE ◽  
...  
Keyword(s):  
Great Lakes ◽  
Water Activity ◽  
Salt Concentration ◽  
Clostridium Botulinum ◽  
Toxin Production ◽  
Water Phase ◽  
Salt Level ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E ◽  
Average Water

The effect of salt level and nitrite on botulinal safety of smoked whitefish was investigated. An average water-phase (wp) salt concentration of 4.4% inhibited outgrowth of Clostridium botulinum type E spores (103 spores/g) for over 35 d in temperature-abused (27°C) smoked whitefish. Incorporation of nitrite (220 mg/kg) during brining to the smoked salted (4.4%, wp) whitefish inhibited toxin production for 56 d at 27°C. An average salt concentration of 6.2% (wp), with or without nitrite, totally inhibited toxin production for the duration of the study (83 d). The effect of pH and water activity in temperature-abused smoked whitefish as a means of controlling toxin production by C. botulinum type E spores was evaluated.

scholarly journals Whole-Genome Single-Nucleotide-Polymorphism Analysis for Discrimination of Clostridium botulinum Group I Strains

2014 ◽  
Vol 80 (7) ◽  
pp. 2125-2132 ◽  
Author(s):  
Narjol Gonzalez-Escalona ◽  
Ruth Timme ◽  
Brian H. Raphael ◽  
Donald Zink ◽  
Shashi K. Sharma
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Clostridium Botulinum ◽  
Toxin Gene ◽  
Polymorphism Analysis ◽  
Snp Analysis ◽  
Whole Genome ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Content Type ◽  
Group I

ABSTRACTClostridium botulinumis a genetically diverse Gram-positive bacterium producing extremely potent neurotoxins (botulinum neurotoxins A through G [BoNT/A-G]). The complete genome sequences of three strains harboring only the BoNT/A1 nucleotide sequence are publicly available. Although these strains contain a toxin cluster (HA+OrfX−) associated with hemagglutinin genes, little is known about the genomes of subtype A1 strains (termed HA−OrfX+) that lack hemagglutinin genes in the toxin gene cluster. We sequenced the genomes of three BoNT/A1-producingC. botulinumstrains: two strains with the HA+OrfX−cluster (69A and 32A) and one strain with the HA−OrfX+cluster (CDC297). Whole-genome phylogenic single-nucleotide-polymorphism (SNP) analysis of these strains along with other publicly availableC. botulinumgroup I strains revealed five distinct lineages. Strains 69A and 32A clustered with theC. botulinumtype A1 Hall group, and strain CDC297 clustered with theC. botulinumtype Ba4 strain 657. This study reports the use of whole-genome SNP sequence analysis for discrimination ofC. botulinumgroup I strains and demonstrates the utility of this analysis in quickly differentiatingC. botulinumstrains harboring identical toxin gene subtypes. This analysis further supports previous work showing that strains CDC297 and 657 likely evolved from a common ancestor and independently acquired separate BoNT/A1 toxin gene clusters at distinct genomic locations.

scholarly journals Role of the Phenylalanine-Hydroxylating System in Aromatic Substance Degradation and Lipid Metabolism in the Oleaginous Fungus Mortierella alpina

2013 ◽  
Vol 79 (10) ◽  
pp. 3225-3233 ◽  
Author(s):  
Hongchao Wang ◽  
Haiqin Chen ◽  
Guangfei Hao ◽  
Bo Yang ◽  
Yun Feng ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
High Performance ◽  
Phenylalanine Hydroxylase ◽  
Uv Spectroscopy ◽  
Dry Weight ◽  
Life Forms ◽  
Mortierella Alpina ◽  
Content Type ◽  
Gene Copies ◽  
Aromatic Substance

ABSTRACTMortierella alpinais a filamentous fungus commonly found in soil that is able to produce lipids in the form of triacylglycerols that account for up to 50% of its dry weight. Analysis of theM. alpinagenome suggests that there is a phenylalanine-hydroxylating system for the catabolism of phenylalanine, which has never been found in fungi before. We characterized the phenylalanine-hydroxylating system inM. alpinato explore its role in phenylalanine metabolism and its relationship to lipid biosynthesis. Significant changes were found in the profile of fatty acids inM. alpinagrown on medium containing an inhibitor of the phenylalanine-hydroxylating system compared toM. alpinagrown on medium without inhibitor. Genes encoding enzymes involved in the phenylalanine-hydroxylating system (phenylalanine hydroxylase [PAH], pterin-4α-carbinolamine dehydratase, and dihydropteridine reductase) were expressed heterologously inEscherichia coli, and the resulting proteins were purified to homogeneity. Their enzymatic activity was investigated by high-performance liquid chromatography (HPLC) or visible (Vis)-UV spectroscopy. Two functional PAH enzymes were observed, encoded by distinct gene copies. A novel role for tetrahydrobiopterin in fungi as a cofactor for PAH, which is similar to its function in higher life forms, is suggested. This study establishes a novel scheme for the fungal degradation of an aromatic substance (phenylalanine) and suggests that the phenylalanine-hydroxylating system is functionally significant in lipid metabolism.

scholarly journals Genetic Diversity among Clostridium botulinum Strains Harboringbont/A2andbont/A3Genes

2012 ◽  
Vol 78 (24) ◽  
pp. 8712-8718 ◽  
Author(s):  
Carolina Lúquez ◽  
Brian H. Raphael ◽  
Lavin A. Joseph ◽  
Sarah R. Meno ◽  
Rafael A. Fernández ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Clostridium Botulinum ◽  
Type A ◽  
Gene Clusters ◽  
Nucleotide Sequencing ◽  
Toxin Gene ◽  
Content Type ◽  
Geographic Origins ◽  
Diversity Studies ◽  
Insight Into

ABSTRACTClostridium botulinumtype A strains are known to be genetically diverse and widespread throughout the world. Genetic diversity studies have focused mainly on strains harboring one type A botulinum toxin gene,bont/A1, although all reportedbont/Agene variants have been associated with botulism cases. Our study provides insight into the genetic diversity ofC. botulinumtype A strains, which containbont/A2(n= 42) andbont/A3(n= 4) genes, isolated from diverse samples and geographic origins. Genetic diversity was assessed by usingbontnucleotide sequencing, content analysis of thebontgene clusters, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Sequences ofbontgenes obtained in this study showed 99.9 to 100% identity with otherbont/A2orbont/A3gene sequences available in public databases. The neurotoxin gene clusters of the subtype A2 and A3 strains analyzed in this study were similar in gene content.C. botulinumstrains harboringbont/A2andbont/A3genes were divided into six and two MLST profiles, respectively. Four groups of strains shared a similarity of at least 95% by PFGE; the largest group included 21 out of 46 strains. The strains analyzed in this study showed relatively limited genetic diversity using either MLST or PFGE.

scholarly journals Distribution of Clostridium botulinum Type E Strains in Nunavik, Northern Quebec, Canada

2012 ◽  
Vol 79 (2) ◽  
pp. 646-654 ◽  
Author(s):  
Daniel Leclair ◽  
Jeffrey M. Farber ◽  
Bill Doidge ◽  
Burke Blanchfield ◽  
Sandy Suppa ◽  
...  
Keyword(s):  
Clostridium Botulinum ◽  
Incidence Rates ◽  
Pulsed Field Gel Electrophoresis ◽  
Hudson Bay ◽  
Multiple Sources ◽  
Content Type ◽  
Intestinal Contents ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E ◽  
Almost All

ABSTRACTThe distribution and levels ofClostridium botulinumtype E were determined from field sites used by Inuit hunters for butchering seals along the coast of Nunavik. The incidence rates ofC. botulinumtype E in shoreline soil along the coast were 0, 50, and 87.5% among samples tested for the Hudson Strait, Hudson Bay, and Ungava Bay regions, respectively. Spores were detected in seawater or coastal rock surfaces from 17.6% of butchering sites, almost all of which were located in southern Ungava Bay. Concentrations ofC. botulinumtype E along the Ungava Bay coast were significantly higher than on the coasts of Hudson Strait and Hudson Bay, with the highest concentrations (270 to 1,800/kg of sample) found near butchering sites located along the mouths of large rivers. The Koksoak River contained high levels ofC. botulinumtype E, with the highest median concentration (270/kg) found in sediments of the marine portion of the river.C. botulinumtype E was found in the intestinal contents (4.4%) and skins (1.4%) of seals. A high genetic biodiversity ofC. botulinumtype E isolates was observed among the 21 butchering sites and their surroundings along the Nunavik coastline, with 83% of isolates (44/53) yielding distinct pulsed-field gel electrophoresis genotypes. Multiple sources ofC. botulinumtype E may be involved in the contamination of seal meat during butchering in this region, but the risk of contamination appears to be much higher from environmental sources along the shoreline of southern Ungava Bay and the sediments of the Koksoak River.

Clostridium botulinum type E occurs and grows in the alga Cladophora glomerata

2009 ◽  
Vol 66 (6) ◽  
pp. 879-882 ◽  
Author(s):  
Muruleedhara N. Byappanahalli ◽  
Richard L. Whitman
Keyword(s):  
Clostridium Perfringens ◽  
Green Algae ◽  
Spore Germination ◽  
Clostridium Botulinum ◽  
Lake Michigan ◽  
Cladophora Glomerata ◽  
Algal Mats ◽  
E Gene ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E

In recent years, massive avian die-offs from Clostridium botulinum type E infection have occurred in the Sleeping Bear Dunes National Lakeshore (SLBE) area of Lake Michigan. These outbreaks have been coincidental with massive blooms of the green algae Cladophora , mostly Cladophora glomerata . We tested the hypothesis that Clostridium botulinum type E can grow under suitable conditions in these algal mats. In a lab mesocosm study, Cladophora from four outbreak-impacted beaches from SLBE were compared with four unimpacted beaches in the Milwaukee–Racine area for bontE gene of Clostridium botulinum. Frequency of the bontE gene was higher after incubation (25 °C for up to 6 weeks) of Cladophora from impacted vs. the unimpacted area. Since no type E gene was detected initially in Cladophora from any of the eight locations, we infer that the increased occurrence of type E gene arose from spore germination or vegetative Clostridium growth within the existing algal mats of SLBE. Moreover, we found that the congener Clostridium perfringens readily grows in mesocosms containing Cladophora.

scholarly journals Two Novel Toxin Variants Revealed by Whole-Genome Sequencing of 175 Clostridium botulinum Type E Strains

2014 ◽  
Vol 80 (20) ◽  
pp. 6334-6345 ◽  
Author(s):  
K. A. Weedmark ◽  
D. L. Lambert ◽  
P. Mabon ◽  
K. L. Hayden ◽  
C. J. Urfano ◽  
...  
Keyword(s):  
Clostridium Botulinum ◽  
Botulinum Neurotoxin ◽  
Hudson Bay ◽  
Whole Genome ◽  
Coding Sequences ◽  
Content Type ◽  
Northern Canada ◽  
River Region ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E

ABSTRACTWe sequenced 175Clostridium botulinumtype E strains isolated from food, clinical, and environmental sources from northern Canada and analyzed their botulinum neurotoxin (bont) coding sequences (CDSs). In addition tobont/E1andbont/E3variant types, neurotoxin sequence analysis identified two novel BoNT type E variants termed E10 and E11. Strains producing type E10 were found along the eastern coastlines of Hudson Bay and the shores of Ungava Bay, while strains producing type E11 were only found in the Koksoak River region of Nunavik. Strains producing BoNT/E3 were widespread throughout northern Canada, with the exception of the coast of eastern Hudson Bay.

Screen Mesh Size Affects Estimates of Macro- and Meio-benthos Abundance and Biomass in the Great Lakes

1981 ◽  
Vol 38 (9) ◽  
pp. 1027-1034 ◽  
Author(s):  
Thomas F. Nalepa ◽  
Andrew Robertson
Keyword(s):  
Great Lakes ◽  
Body Length ◽  
Water Depth ◽  
Lake Michigan ◽  
Mesh Size ◽  
Dry Weight ◽  
Head Capsule ◽  
Future Studies ◽  
Almost All ◽  
Screen Mesh

The efficiencies of screens with mesh openings of 595 and 106 μm in retaining, respectively, the macro- and meio-benthos were measured for samples taken in southeastern Lake Michigan. The use of these screens provides adequate estimates of dry weight biomass for both the macro- and meio-benthos, but serious underestimates of the numbers of many taxa, most notably naidids, enchytraeids, chironomids, nematodes, and rotifers, can result. For chironomids, retention on the 595-μm screen varied by species, with overall retention being closely related to both body length and head capsule width. Retention for certain macro-benthic taxa was significantly related to sampling date and water depth, indicating that future studies concerned with these variables should use mesh sizes small enough to retain all (or almost all) of the individuals of the taxa of interest.Key words: mesh size, screen efficiency, percent retention, macrobenthos, meibenthos, Lake Michigan

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