Last data update: Oct 28, 2024. (Total: 48004 publications since 2009)
Records 1-27 (of 27 Records) |
Query Trace: Kucerova Z[original query] |
---|
Rapid identification of enteric bacteria from whole genome sequences using average nucleotide identity metrics
Lindsey RL , Gladney LM , Huang AD , Griswold T , Katz LS , Dinsmore BA , Im MS , Kucerova Z , Smith PA , Lane C , Carleton HA . Front Microbiol 2023 14 1225207 Identification of enteric bacteria species by whole genome sequence (WGS) analysis requires a rapid and an easily standardized approach. We leveraged the principles of average nucleotide identity using MUMmer (ANIm) software, which calculates the percent bases aligned between two bacterial genomes and their corresponding ANI values, to set threshold values for determining species consistent with the conventional identification methods of known species. The performance of species identification was evaluated using two datasets: the Reference Genome Dataset v2 (RGDv2), consisting of 43 enteric genome assemblies representing 32 species, and the Test Genome Dataset (TGDv1), comprising 454 genome assemblies which is designed to represent all species needed to query for identification, as well as rare and closely related species. The RGDv2 contains six Campylobacter spp., three Escherichia/Shigella spp., one Grimontia hollisae, six Listeria spp., one Photobacterium damselae, two Salmonella spp., and thirteen Vibrio spp., while the TGDv1 contains 454 enteric bacterial genomes representing 42 different species. The analysis showed that, when a standard minimum of 70% genome bases alignment existed, the ANI threshold values determined for these species were ≥95 for Escherichia/Shigella and Vibrio species, ≥93% for Salmonella species, and ≥92% for Campylobacter and Listeria species. Using these metrics, the RGDv2 accurately classified all validation strains in TGDv1 at the species level, which is consistent with the classification based on previous gold standard methods. |
Draft genome sequences of Listeria monocytogenes strains of sequence type 1733 which constituted a pseudo-outbreak due to contaminated blood agar media
Lee S , Sadat A , Kucerova Z , Kathariou S . Microbiol Resour Announc 2023 e0073223 We report genome sequences of Listeria monocytogenes sequence type (ST) 1733 from a 2013 pseudo-outbreak, where L. monocytogenes isolation from non-sterile sites (urine, wound, or abscess) was an artifact from contaminated sheep blood in the isolation media. Two ST1733 strains from wound and urine in 2005 are also reported. |
Draft genome sequences of a historical collection of Listeria monocytogenes from humans and other sources, 1926-1964
Brown P , Murray RGE , Galsworthy S , Ivanova M , Leekitcharoenphon P , Ward T , Kucerova Z , Chen Y , Elhanafi D , Siletzky R , Kathariou S . Microbiol Resour Announc 2023 12 (10) e0062523 Listeria monocytogenes can persistently contaminate food processing environments and tolerate sanitizers. Most sequenced strains are from clinical and environmental sources in the contemporary era, with relatively few prior to extensive food processing and sanitizer use. We report the genome sequences of a diverse panel of 83 strains from 1926 to 1964. |
Corrigendum: Whole Genome Sequencing: Bridging One-Health Surveillance of Foodborne Diseases.
Gerner-Smidt P , Besser J , Concepción-Acevedo J , Folster JP , Huffman J , Joseph LA , Kucerova Z , Nichols MC , Schwensohn CA , Tolar B . Front Public Health 2019 7 365 In the original article, there was a mistake in Figure 1 and Figure 2 as published. The graphics used are different than those originally submitted. |
Horizontal Gene Transfer and Loss of Serotype-Specific Genes in Listeria monocytogenes Can Lead to Incorrect Serotype Designations with a Commonly-Employed Molecular Serotyping Scheme.
Brown P , Kucerova Z , Gorski L , Chen Y , Ivanova M , Leekitcharoenphon P , Parsons C , Niedermeyer J , Jackson J , Kathariou S . Microbiol Spectr 2022 11 (1) e0274522 Listeria monocytogenes is a Gram-positive, facultative intracellular foodborne pathogen capable of causing severe, invasive illness (listeriosis). Three serotypes, 1/2a, 1/2b, and 4b, are leading contributors to human listeriosis, with 4b including the major hypervirulent clones. The multiplex PCR scheme developed by Doumith and collaborators employs primers targeting specific lineages (e.g., lineage II-specific lmo0737, lineage I-specific LMOf2365_2059) or serotypes (e.g., serotype 4b-specific LMOf2365_1900). The Doumith scheme (DS) is extensively employed for molecular serotyping of L. monocytogenes due to its high accuracy, relative ease, and affordability. However, for certain strains, the DS serotype designations are in conflict with those relying on antibody-based schemes or whole-genome sequence (WGS) analysis. In the current study, all 27 tested serotype 4b strains with sequence type 782 (ST782) within the hypervirulent clonal complex 2 (CC2) were designated 1/2b/3b using the DS. These strains lacked the serotype 4b-specific gene LMOf2365_1900, while retaining LMOf2365_2059, which, together with prs, yields the DS 1/2b/3b profile. Furthermore, 15 serotype 1/2a strains of four STs, mostly from water, were designated 1/2b/3b using the DS. These strains lacked the lmo0737 cassette but harbored genomic islands with LMOf2365_2059, thus yielding the DS 1/2b/3b profile. Lastly, we investigated a novel, dual 1/2a-1/2b profile obtained using the DS with 21 serotype 1/2a strains of four STs harboring both the lmo0737 cassette and genomic islands with LMOf2365_2059. The findings suggest that for certain strains and clones of L. monocytogenes the DS designations should be viewed with caution and complemented with alternative tools, e.g., traditional serotyping or WGS analysis. IMPORTANCE Listeria monocytogenes is a foodborne pathogen responsible for severe illness (listeriosis), especially in pregnant women and their fetuses, immunocompromised individuals, and the elderly. Three serotypes, 1/2a, 1/2b, and 4b, account for most human listeriosis, with certain serotype 4b clonal complexes (CCs) overrepresented in human disease. Serotyping remains extensively employed in Listeria epidemiologic investigations, and a multiplex PCR-based serotyping scheme is widely used. However, the PCR gene targets can be lost or gained via horizontal gene transfer, leading to novel PCR profiles without known serotype designations or to incorrect serotype assignments. Thus, an entire serotype 4b clone of the hypervirulent CC2 would be misidentified as serotype 1/2b, and several strains of serotype 1/2a would be identified as serotype 1/2b. Such challenges are especially common in novel clones from underexplored habitats, e.g., wildlife and surface water. The findings suggest caution in application of molecular serotyping, while highlighting Listeria's diversity and potential for horizontal gene transfer. |
Genome Sequences of Hemolytic and Nonhemolytic Listeria innocua Strains from Human, Food, and Environmental Sources.
McIntosh T , Kucerova Z , Katz LS , Lilley CM , Rowe LA , Unoarumhi Y , Batra D , Burnett E , Smikle M , Lee C . Microbiol Resour Announc 2022 11 (12) e0072322 This report describes genome sequences for nine Listeria innocua strains that varied in hemolytic phenotypes on sheep blood agar. All strains were sequenced using Pacific Biosciences (PacBio) single-molecule real-time (SMRT) chemistry; overall, the average read length of these sequences was 2,869,880 bp, with an average GC content of 37%. |
Listeria monocytogenes Illness and Deaths Associated With Ongoing Contamination of a Multi-Regional Brand of Ice Cream Products, United States, 2010-2015.
Conrad AR , Tubach S , Cantu V , Webb LM , Stroika S , Moris S , Davis M , Hunt DC , Bradley KK , Kucerova Z , Strain E , Doyle M , Fields A , Neil KP , Gould LH , Jackson KA , Wise ME , Griffin PM , Jackson BR . Clin Infect Dis 2022 76 (1) 89-95 BACKGROUND: Frozen foods have rarely been linked to Listeria monocytogenes illness. We describe an outbreak investigation prompted both by hospital clustering of illnesses and product testing. METHODS: We identified outbreak-associated listeriosis cases using whole-genome sequencing (WGS), product testing results, and epidemiologic linkage to cases in the same Kansas hospital. We reviewed hospital medical and dietary records, product invoices, and molecular subtyping results. Federal and state officials tested product and environmental samples for L. monocytogenes. RESULTS: Kansas officials were investigating five cases of listeriosis at a single hospital when, simultaneously, unrelated sampling for a study in South Carolina identified L. monocytogenes in Company A ice cream products made in Texas. Isolates from four patients and Company A products were closely related by WGS, and the four patients with known exposures had consumed milkshakes made with Company A ice cream while hospitalized. Further testing identified L. monocytogenes in ice cream produced in a second Company A production facility in Oklahoma; these isolates were closely related by WGS to those from five patients in three other states. These ten illnesses, involving three deaths, occurred from 2010 through 2015. Company A ultimately recalled all products. CONCLUSION: In this U.S. outbreak of listeriosis linked to a widely distributed brand of ice cream, WGS and product sampling helped link cases spanning five years to two production facilities, indicating longstanding contamination. Comprehensive sanitation controls and environmental and product testing for L. monocytogenes, with regulatory oversight, should be implemented for ice cream production. |
Whole-Genome Sequencing Reveals Multiple Subpopulations of Dominant and Persistent Lineage I Isolates of Listeria monocytogenes in Two Meat Processing Facilities during 2011-2015.
Burnett E , Kucerova Z , Freeman M , Kathariou S , Chen J , Smikle M . Microorganisms 2022 10 (5) Listeria monocytogenes is a foodborne pathogen with a highly clonal population structure comprising multiple phylogenetic sub-groups that can persist within food processing environments and contaminate food. The epidemiology of L. monocytogenes is well-described in some developed countries; however, little is known about the prevalence and population structure of this pathogen in food and food processing environments located in less developed regions. The aim of this study was to determine the genetic characteristics and clonal relatedness of L. monocytogenes that were isolated from two Jamaican meat processing facilities. Of the 37 isolates collected between 2011 and 2015, only a single lineage II isolate was recovered (serotype 1/2c), and the remaining were lineage I isolates representing serotypes 4b, 1/2b, 3b, and two untypeable isolates. Pulsed-field gel electrophoresis (PFGE) delineated isolates into seven pulsotypes, and whole-genome sequencing (WGS) categorized most isolates within one of three clonal complexes (CC): CC2 (N = 12), CC5 (N = 11), and CC288 (N = 11). Isolates representing CC1 (N = 2) and CC9 (N = 1) were also recovered. Virulence-associated genes such as inlA and the LIPI-3 cluster were detected in multiple isolates, along with the stress survival islet cluster-1 (SSI-1), and benzalkonium (bcrABC) and cadmium (cad1, cad2, cad4) resistance cassettes. Multiple isolates that belong to the same CC and matching PFGE patterns were isolated from food and the environment from both facilities across multiple years, suggesting the presence of persistent strains of L. monocytogenes, and/or constant re-entry of the pathogens into the facilities from common sources. These findings highlight the ability of lineage I isolates of L. monocytogenes to colonize, persist, and predominate within two meat-producing environments, and underscores the need for robust surveillance strategies to monitor and mitigate against these important foodborne pathogens. |
Septicaemic Listeriosis in a White-Faced Saki (Pithecia pithecia).
Struthers JD , Kucerova Z , Finley A , Goe A , Huffman J , Phair K . J Comp Pathol 2022 194 7-13 A 27-year-old female white-faced saki (Pithecia pithecia) died following an onset of vomiting and ptyalism. Necropsy revealed lesions of suppurative ventriculitis, choroid plexitis, periventricular encephalitis and meningitis with intralesional gram-positive coccobacilli and paired rods. The saki also had suppurative to mononuclear hepatitis, mild intestinal crypt necrosis, proliferative glomerulonephritis, aortic arteriosclerosis, pulmonary interstitial fibrosis, chronic mild epicarditis, ovarian medullary arteriopathy and a focal superficial cerebral fibrotic nodule with surrounding chronic mixed cell inflammation. Listeria monocytogenes was cultured from liver and spinal cord. Intralesional Listeria bacteria were immunolabelled in brain sections and real-time polymerase chain reaction of brain tissue detected L. monocytogenes. Whole genome multilocus sequence typing characterized the cultured bacterial isolates as sequence type 6 and clonal complex 6. A database search for related clinical and food listerial outbreaks identified genetically related isolates but, because these isolates were more than 20 alleles distant from the saki isolates, they were not a related cluster. Reports of listeriosis in non-human primates are infrequent, and when infections do occur, they tend to be haematogenous with the propensity to cause meningoencephalitis. This saki likely ingested environmental L. monocytogenes, which resulted in disease that may have been facilitated by pre-existing co-morbidities and age. © 2022 Elsevier Ltd |
Genome Sequences of Neurotropic Lineage III Listeria monocytogenes Isolates UKVDL9 and 2010L-2198.
Albrecht TM , Kucerova Z , D'Orazio SEF . Microbiol Resour Announc 2021 10 (18) We report the whole-genome sequence of Listeria monocytogenes UKVDL9 and an edited draft genome sequence of L. monocytogenes 2010L-2198. Both are neurotropic lineage III strains; UKVDL9 was isolated from a sheep brain, and 2010L-2198 was isolated from a human subject with rhombencephalitis. |
Complete Genome Sequence of a Serotype 7 Listeria monocytogenes Strain, FSL R9-0915.
Peters TL , Hudson LK , Bryan DW , Song Y , den Bakker HC , Kucerova Z , Denes TG . Microbiol Resour Announc 2021 10 (1) Listeria monocytogenes serotype 7 lacks glycosidic constituents in wall teichoic acids. Here, we present the complete genome sequence of L. monocytogenes serotype 7 strain FSL R9-0915 and an analysis of genes known to affect L. monocytogenes antigenicity. This strain is used as a control strain in Listeria phage host range analyses. |
Whole Genome Sequencing: Bridging One-Health Surveillance of Foodborne Diseases.
Gerner-Smidt P , Besser J , Concepcion-Acevedo J , Folster JP , Huffman J , Joseph LA , Kucerova Z , Nichols MC , Schwensohn CA , Tolar B . Front Public Health 2019 7 172 Infections caused by pathogens commonly acquired from consumption of food are not always transmitted by that route. They may also be transmitted through contact to animals, other humans or the environment. Additionally, many outbreaks are associated with food contaminated from these non-food sources. For this reason, such presumed foodborne outbreaks are best investigated through a One Health approach working across human, animal and environmental sectors and disciplines. Outbreak strains or clones that have propagated and continue to evolve in non-human sources and environments often show more sequence variation than observed in typical monoclonal point-source outbreaks. This represents a challenge when using whole genome sequencing (WGS), the new gold standard for molecular surveillance of foodborne pathogens, for outbreak detection and investigation. In this review, using recent examples from outbreaks investigated in the United States (US) some aspects of One Health approaches that have been used successfully to solve such outbreaks are presented. These include using different combinations of flexible WGS based case definition, efficient epidemiological follow-up, traceback, surveillance, and testing of potential food and environmental sources and animal hosts. |
Listeriosis outbreaks associated with soft cheeses, United States, 1998-2014
Jackson KA , Gould LH , Hunter JC , Kucerova Z , Jackson B . Emerg Infect Dis 2018 24 (6) 1116-1118 Since 2006, the number of reported US listeriosis outbreaks associated with cheese made under unsanitary conditions has increased. Two-thirds were linked to Latin-style soft cheese, often affecting pregnant Hispanic women and their newborns. Adherence to pasteurization protocols and sanitation measures to avoid contamination after pasteurization can reduce future outbreaks. |
Multistate outbreak of Listeria monocytogenes infections linked to whole apples used in commercially produced, prepackaged caramel apples: United States, 2014-2015.
Angelo KM , Conrad AR , Saupe A , Dragoo H , West N , Sorenson A , Barnes A , Doyle M , Beal J , Jackson KA , Stroika S , Tarr C , Kucerova Z , Lance S , Gould LH , Wise M , Jackson BR . Epidemiol Infect 2017 145 (5) 1-9 Whole apples have not been previously implicated in outbreaks of foodborne bacterial illness. We investigated a nationwide listeriosis outbreak associated with caramel apples. We defined an outbreak-associated case as an infection with one or both of two outbreak strains of Listeria monocytogenes highly related by whole-genome multilocus sequence typing (wgMLST) from 1 October 2014 to 1 February 2015. Single-interviewer open-ended interviews identified the source. Outbreak-associated cases were compared with non-outbreak-associated cases and traceback and environmental investigations were performed. We identified 35 outbreak-associated cases in 12 states; 34 (97%) were hospitalized and seven (20%) died. Outbreak-associated ill persons were more likely to have eaten commercially produced, prepackaged caramel apples (odds ratio 326.7, 95% confidence interval 32.2-3314). Environmental samples from the grower's packing facility and distribution-chain whole apples yielded isolates highly related to outbreak isolates by wgMLST. This outbreak highlights the importance of minimizing produce contamination with L. monocytogenes. Investigators should perform single-interviewer open-ended interviews when a food is not readily identified. |
Whole genome-based population biology and epidemiological surveillance of Listeria monocytogenes.
Moura A , Criscuolo A , Pouseele H , Maury MM , Leclercq A , Tarr C , Bjorkman JT , Dallman T , Reimer A , Enouf V , Larsonneur E , Carleton H , Bracq-Dieye H , Katz LS , Jones L , Touchon M , Tourdjman M , Walker M , Stroika S , Cantinelli T , Chenal-Francisque V , Kucerova Z , Rocha EP , Nadon C , Grant K , Nielsen EM , Pot B , Gerner-Smidt P , Lecuit M , Brisse S . Nat Microbiol 2016 2 16185 Listeria monocytogenes (Lm) is a major human foodborne pathogen. Numerous Lm outbreaks have been reported worldwide and associated with a high case fatality rate, reinforcing the need for strongly coordinated surveillance and outbreak control. We developed a universally applicable genome-wide strain genotyping approach and investigated the population diversity of Lm using 1,696 isolates from diverse sources and geographical locations. We define, with unprecedented precision, the population structure of Lm, demonstrate the occurrence of international circulation of strains and reveal the extent of heterogeneity in virulence and stress resistance genomic features among clinical and food isolates. Using historical isolates, we show that the evolutionary rate of Lm from lineage I and lineage II is low ( approximately 2.5 x 10-7 substitutions per site per year, as inferred from the core genome) and that major sublineages (corresponding to so-called 'epidemic clones') are estimated to be at least 50-150 years old. This work demonstrates the urgent need to monitor Lm strains at the global level and provides the unified approach needed for global harmonization of Lm genome-based typing and population biology. |
Implementation of Nationwide Real-time Whole-genome Sequencing to Enhance Listeriosis Outbreak Detection and Investigation.
Jackson BR , Tarr C , Strain E , Jackson KA , Conrad A , Carleton H , Katz LS , Stroika S , Gould LH , Mody RK , Silk BJ , Beal J , Chen Y , Timme R , Doyle M , Fields A , Wise M , Tillman G , Defibaugh-Chavez S , Kucerova Z , Sabol A , Roache K , Trees E , Simmons M , Wasilenko J , Kubota K , Pouseele H , Klimke W , Besser J , Brown E , Allard M , Gerner-Smidt P . Clin Infect Dis 2016 63 (3) 380-6 Listeria monocytogenes(Lm) causes severe foodborne illness (listeriosis). Previous molecular subtyping methods, such as pulsed-field gel electrophoresis (PFGE), were critical in detecting outbreaks that led to food safety improvements and declining incidence, but PFGE provides limited genetic resolution. A multiagency collaboration began performing real-time, whole-genome sequencing (WGS) on all U.S.Lmisolates from patients, food, and the environment in September 2013, posting sequencing data into a public repository. Compared with the year before the project began, WGS, combined with epidemiologic and product trace-back data, detected more listeriosis clusters and solved more outbreaks (2 outbreaks in pre-WGS year, 5 in WGS year 1, and 9 in year 2). Whole-genome multilocus sequence typing and single nucleotide polymorphism analyses provided equivalent phylogenetic relationships relevant to investigations; results were most useful when interpreted in context of epidemiological data. WGS has transformed listeriosis outbreak surveillance and is being implemented for other foodborne pathogens. |
Two Listeria monocytogenes pseudo-outbreaks caused by contaminated laboratory culture media
Matanock A , Katz LS , Jackson KA , Kucerova Z , Conrad AR , Glover WA , Nguyen V , Mohr MC , Marsden-Haug N , Thompson D , Dunn JR , Stroika S , Melius B , Tarr C , Dietrich SE , Kao AS , Kornstein L , Li Z , Maroufi A , Marder EP , Meyer R , Perez-Osorio AC , Reddy V , Reporter R , Carleton H , Tweeten S , Waechter H , Yee LM , Wise ME , Davis K , Jackson B . J Clin Microbiol 2015 54 (3) 768-70 Listeriosis is a serious foodborne infection that disproportionately affects elderly adults, pregnant women, newborns, and immunocompromised individuals. Diagnosis is made by culturing Listeria monocytogenes from sterile body fluids or products of conception. This report describes investigations of two listeriosis pseudo-outbreaks caused by contaminated laboratory media made from sheep blood. |
Evolutionary Relationships of Outbreak-associated Listeria monocytogenes Strains of Serotypes 1/2a and 1/2b Determined by Whole Genome Sequencing.
Bergholz TM , den Bakker HC , Katz LS , Silk BJ , Jackson KA , Kucerova Z , Joseph LA , Turnsek M , Gladney LM , Halpin JL , Xavier K , Gossack J , Ward TJ , Frace M , Tarr CL . Appl Environ Microbiol 2015 82 (3) 928-38 We used whole genome sequencing to determine evolutionary relationships among 20 outbreak-associated clinical isolates of Listeria monocytogenes serotypes 1/2a and 1/2b. Isolates from six of eleven outbreaks fell outside of the clonal groups or 'epidemic clones' that have been previously associated with outbreaks, suggesting that epidemic potential may be widespread in L. monocytogenes and is not limited to the recognized epidemic clones. Pairwise comparisons between epidemiologically-related isolates within clonal complexes showed that genome-level variation differed by two orders of magnitude between different comparisons, and the distribution of point mutations (core versus accessory genome) also varied. In addition, genetic divergence between one closely related pair of isolates from a single outbreak was driven primarily by changes in phage regions. The evolutionary analysis showed the changes could be attributed to horizontal gene transfer; members of the diverse bacterial community found in the production facility could have served as the source of novel genetic material at some point in the production chain. The results raise the question of how to best utilize information contained within the accessory genome in outbreak investigations. The full magnitude and complexity of genetic changes revealed by genome sequencing could not be discerned from traditional subtyping methods and the results demonstrate the challenges of interpreting genetic variation among isolates recovered from a single outbreak. Epidemiological information remains critical for proper interpretation of nucleotide and structural diversity among isolates recovered during outbreaks, and will remain so until we understand more about how various population histories influence genetic variation. |
Predominance and distribution of a persistent Listeria monocytogenes clone in a commercial fresh mushroom processing environment
Murugesan L , Kucerova Z , Knabel SJ , LaBorde LF . J Food Prot 2015 78 (11) 1988-98 A longitudinal study was conducted to determine the prevalence of Listeria spp. in a commercial fresh mushroom slicing and packaging environment. Samples were collected at three different sampling periods within a 13-month time interval. Of the 255 environmental samples collected, 18.8% tested positive for L. monocytogenes, 4.3% for L. innocua, and 2.0% for L. grayi. L. monocytogenes was most often found on wet floors within the washing and slicing and packaging areas. Each of the 171 L. monocytogenes isolates found in the environment could be placed into one of three different serotypes; 1/2c was predominant (93.6%), followed by 1/2b (3.5%) and 1/2a (2.9%). Of 58 isolates subtyped using multi-virulence-locus sequence typing, all 1/2c isolates were identified as virulence type (VT) 11 (VT11), all 1/2b isolates were VT105, and 1/2a isolates were either VT107 or VT56. VT11 was designated as the predominant and persistent clone in the environment because it was isolated repeatedly at numerous locations throughout the study. The overall predominance and persistence of VT11 indicates that it likely colonized the mushroom processing environment. Areas adjacent to the trench drain in the washing and slicing area and a floor crack in the packaging area may represent primary harborage sites (reservoirs) for VT11. Improvements made to sanitation procedures by company management after period 2 coincided with a significant (P ≤ 0.001) reduction in the prevalence of L. monocytogenes from 17.8% in period 1 and 30.7% in period 2 to 8.5% in period 3. This suggests that targeted cleaning and sanitizing procedures can be effective in minimizing the occurrence of L. monocytogenes contamination in processing facilities. Additional research is needed to understand why VT11 was predominant and persistent in the mushroom processing environment. |
Assessment of blood-brain barrier penetration of miltefosine used to treat a fatal case of granulomatous amebic encephalitis possibly caused by an unusual Balamuthia mandrillaris strain
Roy SL , Atkins JT , Gennuso R , Kofos D , Sriram RR , Dorlo TP , Hayes T , Qvarnstrom Y , Kucerova Z , Guglielmo BJ , Visvesvara GS . Parasitol Res 2015 114 (12) 4431-9 Balamuthia mandrillaris, a free-living ameba, causes rare but frequently fatal granulomatous amebic encephalitis (GAE). Few patients have survived after receiving experimental drug combinations, with or without brain lesion excisions. Some GAE survivors have been treated with a multi-drug regimen including miltefosine, an investigational anti-leishmanial agent with in vitro amebacidal activity. Miltefosine dosing for GAE has been based on leishmaniasis dosing because no data exist in humans concerning its pharmacologic distribution in the central nervous system. We describe results of limited cerebrospinal fluid (CSF) and serum drug level testing performed during clinical management of a child with fatal GAE who was treated with a multiple drug regimen including miltefosine. Brain biopsy specimens, CSF, and sera were tested for B. mandrillaris using multiple techniques, including culture, real-time polymerase chain reaction, immunohistochemical techniques, and serology. CSF and serum miltefosine levels were determined using a liquid chromatography method coupled to tandem mass spectrometry. The CSF miltefosine concentration on hospital admission day 12 was 0.4 mug/mL. The serum miltefosine concentration on day 37, about 80 h post-miltefosine treatment, was 15.3 mug/mL. These are the first results confirming some blood-brain barrier penetration by miltefosine in a human, although with low-level CSF accumulation. Further evaluation of brain parenchyma penetration is required to determine optimal miltefosine dosing for Balamuthia GAE, balanced with the drug's toxicity profile. Additionally, the Balamuthia isolate was evaluated by real-time polymerase chain reaction (PCR), demonstrating genetic variability in 18S ribosomal RNA (18S rRNA) sequences and possibly signaling the first identification of multiple Balamuthia strains with varying pathogenicities. |
Multistate outbreak of listeriosis caused by imported cheese and evidence of cross-contamination of other cheeses, USA, 2012
Heiman KE , Garalde VB , Gronostaj M , Jackson KA , Beam S , Joseph L , Saupe A , Ricotta E , Waechter H , Wellman A , Adams-Cameron M , Ray G , Fields A , Chen Y , Datta A , Burall L , Sabol A , Kucerova Z , Trees E , Metz M , Leblanc P , Lance S , Griffin PM , Tauxe RV , Silk BJ . Epidemiol Infect 2015 144 (13) 1-11 Listeria monocytogenes is a foodborne pathogen that can cause bacteraemia, meningitis, and complications during pregnancy. In July 2012, molecular subtyping identified indistinguishable L. monocytogenes isolates from six patients and two samples of different cut and repackaged cheeses. A multistate outbreak investigation was initiated. Initial analyses identified an association between eating soft cheese and outbreak-related illness (odds ratio 17.3, 95% confidence interval 2.0-825.7) but no common brand. Cheese inventory data from locations where patients bought cheese and an additional location where repackaged cheese yielded the outbreak strain were compared to identify cheeses for microbiological sampling. Intact packages of imported ricotta salata yielded the outbreak strain. Fourteen jurisdictions reported 22 cases from March-October 2012, including four deaths and a fetal loss. Six patients ultimately reported eating ricotta salata; another reported eating cheese likely cut with equipment also used for contaminated ricotta salata, and nine more reported eating other cheeses that might also have been cross-contaminated. An FDA import alert and US and international recalls followed. Epidemiology-directed microbiological testing of suspect cheeses helped identify the outbreak source. Cross-contamination of cheese highlights the importance of using validated disinfectant protocols and routine cleaning and sanitizing after cutting each block or wheel. |
Notes from the field: listeriosis associated with stone fruit - United States, 2014
Jackson BR , Salter M , Tarr C , Conrad A , Harvey E , Steinbock L , Saupe A , Sorenson A , Katz L , Stroika S , Jackson KA , Carleton H , Kucerova Z , Melka D , Strain E , Parish M , Mody RK . MMWR Morb Mortal Wkly Rep 2015 64 (10) 282-3 On July 19, 2014, a packing company in California (company A) voluntarily recalled certain lots of stone fruits, including whole peaches, nectarines, plums, and pluots, because of concern about contamination with Listeria monocytogenes based on internal company testing. On July 31, the recall was expanded to cover all fruit packed at their facility during June 1-July 17. After the initial recall, clinicians, state and local health departments, CDC, and the Food and Drug Administration (FDA) received many inquiries about listeriosis from concerned consumers, many of whom had received automated telephone calls informing them that they had purchased recalled fruit. During July 19-31, the CDC Listeria website received >500,000 page views, more than seven times the views received during the previous 52 weeks. However, no molecular information from L. monocytogenes isolates was available to assess whether human illnesses might be linked to these products. |
Serologic survey for exposure following fatal Balamuthia mandrillaris infection
Jackson BR , Kucerova Z , Roy SL , Aguirre G , Weiss J , Sriram R , Yoder J , Foelber R , Baty S , Derado G , Stramer SL , Winkelman V , Visvesvara GS . Parasitol Res 2014 113 (4) 1305-11 Granulomatous amebic encephalitis (GAE) from Balamuthia mandrillaris, a free-living ameba, has a case fatality rate exceeding 90 % among recognized cases in the USA. In August 2010, a GAE cluster occurred following transplantation of infected organs from a previously healthy landscaper in Tucson, AZ, USA, who died from a suspected stroke. As B. mandrillaris is thought to be transmitted through soil, a serologic survey of landscapers and a comparison group of blood donors in southern Arizona was performed. Three (3.6 %) of 83 serum samples from landscapers and 11 (2.5 %) of 441 serum samples from blood donors were seropositive (p = 0.47). On multivariable analysis, county of residence was associated with seropositivity, whereas age, sex, and ethnicity were not. Exposure to B. mandrillaris, previously unexamined in North America, appears to be far more common than GAE in Southern Arizona. Risk factors for disease progression and the ameba's geographic range should be examined. |
Multiplex bead assay for serum samples from children in Haiti enrolled in a drug study for the treatment of lymphatic filariasis
Moss DM , Priest JW , Boyd A , Weinkopff T , Kucerova Z , Beach MJ , Lammie PJ . Am J Trop Med Hyg 2011 85 (2) 229-37 A multiplex bead assay (MBA) was used to analyze serum samples collected longitudinally from children enrolled in a drug trial for treatment of filariasis in Leogane, Haiti. Recombinant antigens Bm14 and Bm33 from Brugia malayi, third polar tube protein (PTP3) from Encephalitozoon cuniculi, and merozoite surface protein-1(19) (MSP-1(19)) from Plasmodium falciparum were coupled to carboxylated polystyrene microspheres. IgG responses to PTP3 and MSP-1(19) were not affected by albendazole (ALB), diethylcarbamazine (DEC), or combination of diethylcarbamazine and albendazole (DEC/ALB). However, IgG and IgG4 responses to Bm14 and Bm33 were significantly decreased (P < 0.001) by DEC and DEC/ALB treatment. Antibody responses to Bm14 and Bm33 decreased after DEC treatment (but not placebo) among children who were negative for microfilaremia and antigenemia at baseline, suggesting that these children harbored early stages of infection. The MBA is an excellent serologic technique for multiple antigens that offers substantial advantages over single-antigen based enzyme-linked immunosorbent assay in mass drug administration studies for monitoring changes in antibody levels. |
Identification of antigenic targets for immunodetection of Balamuthia mandrillaris infection
Kucerova Z , Sriram R , Wilkins PP , Visvesvara GS . Clin Vaccine Immunol 2011 18 (8) 1297-301 The free-living amoeba Balamuthia mandrillaris causes granulomatous amoebic encephalitis (GAE) in humans. Rapid identification of balamuthiasis is critical for effective therapeutic intervention and case management. In the present study we identified target antigens for development of a serologic assay for B. mandrillaris infection. We demonstrated by silver staining that protein profiles for all eight isolates of B. mandrillaris, independent of human or animal origin or geographic origin, appeared to be similar except for some minor differences, indicating the molecular homogeneity of these strains. The profiles of all isolates ranged from 200 to 10 kDa were similar with a prominent protein visible around 30 kDa; all appeared considerably different from protein profiles of the control E6 cells, Acanthamoeba, and Naegleria isolates. Western blot analysis with rabbit hyperimmune serum identified the major immunodominant antigens of 25, 50, 75 and 80 kDa; positive human sera reacted strongly with proteins around 25, 40, 50 and 75 kDa. Proteins around 40 kDa detected by human serum were not recognized by hyperimmune rabbit serum. None of the target proteins were detected by uninfected control sera. Reactivities of five patients sera with 4 different isolates of B. mandrillaris (two strains of human and two strains of animal origins) revealed that patients' sera reacted slightly differently with different B. mandrillaris isolates, although major proteins of approximately 25, 50 and 75kDa were present in all extracts. |
Microsporidiosis and cryptosporidiosis in HIV/AIDS patients in St. Petersburg, Russia: serological identification of microsporidia and Cryptosporidium parvum in sera samples from HIV/AIDS patients
Kucerova Z , Sokolova OI , Demyanov AV , Kvac M , Sak B , Kvetonova D , Secor WE . AIDS Res Hum Retroviruses 2010 27 (1) 13-5 To determine seroprevalence of the opportunistic organisms Cryptosporidium parvum and microsporidia (Encephalitozoon cuniculi, E. intestinalis, E. hellem, and Enterocytozoon bieneusi) in Russian HIV/AIDS patients, we evaluated 46 sera from HIV/AIDS patients from the S.P. Botkin Clinical Infectious Diseases Hospital, St. Petersburg, Russia. Five (10.9%) sera were seropositive for E. cuniculi and 19 (41.3%) were positive for C. parvum by ELISA. By IFAT, 6 (13.0%) sera were seropositive for E. bieneusi, 4 (8.7%) for E. intestinalis, and 9 (19.6%) for E. hellem. This study is the first report to estimate the prevalence of infection with Cryptosporidium and microsporidia among Russian HIV/AIDS patients. |
Seropositivity for Enterocytozoon bieneusi, Czech Republic
Sak B , Kucerova Z , Kvac M , Kvetonova D , Rost M , Secor EW . Emerg Infect Dis 2010 16 (2) 335-7 To determine seropositivity for Enterocytozoon bieneusi in the Czech Republic, we tested 115 serum samples from various groups. We found that 20% from HIV-positive persons, 33% from persons with occupational exposure to animals, and 10% from healthy persons were positive by indirect immunofluorescence assay. Proteins of 32 kDa were detected in serum samples from seropositive persons. |
- Page last reviewed:Feb 1, 2024
- Page last updated:Oct 28, 2024
- Content source:
- Powered by CDC PHGKB Infrastructure