Multilocus Genetic Characterization of Two Ant Vectors (Group II “Dirty 22” Species) Known To Contaminate Food and Food Products and Spread Foodborne Pathogens†

2012 ◽  
Vol 75 (8) ◽  
pp. 1447-1452 ◽  
Author(s):  
IRSHAD M. SULAIMAN ◽  
MICKEY ANDERSON ◽  
DAVID H. OI ◽  
STEVEN SIMPSON ◽  
KHALIL KERDAHI
Keyword(s):  
Foodborne Pathogens ◽  
Nested Pcr ◽  
Genetic Characterization ◽  
Food Products ◽  
Species Group ◽  
Pest Species ◽  
Group I ◽  
Group Ii ◽  
Primer Sets ◽  
Pcr Primer

The U.S. Food and Drug Administration utilizes the presence of filth and extraneous materials as one of the criteria for implementing regulatory actions and assessing adulteration of food products of public health importance. Twenty-two prevalent pest species (also known as the “Dirty 22” species) have been considered by this agency as possible vehicles for the spread of foodborne diseases, and the presence of these species is considered an indicator of unsanitary conditions in food processing and storage facilities. In a previous study, we further categorized the Dirty 22 species into four groups: group I includes four cockroach species, group II includes two ant species, group III includes 12 fly species, and group IV includes four rodent species. Here, we describe the development of three nested PCR primer sets and multilocus genetic characterization by amplifying the small subunit rRNA, elongation factor 1-alpha, and wingless (WNT-1) genes of group II Dirty 22 ant species Monomorium pharaonis and Solenopsis molesta. These novel group II Dirty 22 species-specific nested PCR primer sets can be used when the specimens cannot be identified using conventional microscopic methods. These newly developed assays will provide correct identification of group II Dirty 22 ant species, and the information can be used in the control of foodborne pathogens.

Identification of 18 vector species belonging to Group I, Group II, and Group III ‘Dirty 22’ species known to contaminate food and spread foodborne pathogens: DNA barcoding study of public health importance

2016 ◽  
Vol 37 (01) ◽  
pp. 1-10 ◽  
Author(s):  
Irshad M. Sulaiman ◽  
Emily Jacobs ◽  
Steven Simpson ◽  
Khalil Kerdahi
Keyword(s):  
Public Health ◽  
Foodborne Pathogens ◽  
Species Group ◽  
Diagnostic Methods ◽  
Coi Gene ◽  
Public Health Importance ◽  
Group Iii ◽  
Group I ◽  
Coi Barcoding ◽  
Group Ii

AbstractThe US Food and Drug Administration (US-FDA) uses the presence of filth and extraneous materials as one of the criteria in implementing regulatory actions and assessing food adulteration of public health importance. So far, 22 common pest species (‘Dirty 22’ species) have been considered by this agency for the spreading of foodborne illness, and their presence is an indicator of unsanitary conditions in food processing and storage facilities. Recently, we classified the ‘Dirty 22’ species into four groups: Group I (four cockroach species), Group II (two ant species), Group III (12 fly species), and Group IV (four rodent species), and described two molecular diagnostic methods for group-specific identification. We developed a PCR-RFLP assay based on rRNA gene for the detection and differentiation of Group I ‘Dirty 22’ species. Later, we designed three Group II ‘Dirty 22’ species-specific nested PCR primer sets and sequence characterized the rRNA, elongation factor 1-alpha (EF-1a), and wingless (WNT-1) loci. In this follow-up study, we have evaluated the robustness of five unique sets of published primers targeting the mitochondrial cytochrome oxidase I (COI) gene for insect barcoding. With modified PCR conditions, we successfully used COI barcoding for 18 members of Group I, Group II, and Group III ‘Dirty 22’ species. Results of this study reveal that COI barcoding is an effective tool for rapid identification of insects of Groups I, II, and III ‘Dirty 22’ species known to contaminate food and spread foodborne pathogens.

Droplet Digital PCR Reliably Detects a Single Copy of BCR-ABL1

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2784-2784
Author(s):  
Jaspal Kaeda ◽  
Simone Bonecker ◽  
Frauke Ringel ◽  
Michaela Schwarz ◽  
Bernd Dörken ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Chronic Phase ◽  
Digital Pcr ◽  
Group Iv ◽  
Droplet Digital Pcr ◽  
Pcr Assay ◽  
Group Iii ◽  
Group I ◽  
Increased Sensitivity ◽  
Group Ii

Abstract Data show 40% of chronic myeloid leukemia (CML) patients maintain complete molecular remission (CMR), i.e. failure to detect BCR-ABL1, consenting to termination of Imatinib mesylate (IM) therapy, following undetectable disease for ≥2 years by quantitative PCR (Q-PCR). These findings suggest majority of the patients experience molecular relapse. Furthermore, majority relapse in the first 6 months, implying Q-PCR assay sensitivity is suboptimum, to confidently identify patients for discontinuation of IM. Droplet digital PCR (ddPCR) is suggested to have sensitivity that is one log greater than the Taqman (Q-PCR) assay. If verified, ddPCR would enhance safe withdrawal of IM therapy from CML patients. Here we present data comparing ddPCR with Q-PCR. In total we assayed 161 samples, of these 6 were serial dilutions of the International reference (IR) BCR-ABL1 plasmid, the remainder were cDNA samples. The 161 samples comprised of 4 sample groups; I: CML chronic phase samples (n=118); II: CML samples post stem cell transplant (SCT) (n=22); III: normal control (NC) samples (n=16); IV: Serially diluted BCR-ABL1 1.04x10e2 copies/µl, IR ERM-AD623e (n=6). Group I comprised of 121 samples from 21 CML patients in chronic phase treated with IM. Group II comprised of 21 samples from 19 CML patients (2 samples each for 2 of these 19 patients) who had undergone SCT. Group II comprised of samples from normal adult blood donor volunteers. Finally, Group IV included 6 serially diluted samples, ranging 100 to 0.001 copies of the IR (1.04x10e2 copies/µl) plasmid (Sigma, Munich, Germany). All the Group I, III and IV samples were subjected to Q-PCR Taqman assay and ddPCR (Biorad, California, USA). The Group III samples were in addition subjected to nested PCR. Only those samples with cycle threshold (Ct) <37 in 2 or more replicates by Q-PCR were recorded positive. For ddPCR those samples with sum of ≥3 positive droplets with a minimum 5500 droplets per well were reported positive. All the PCR reactions were performed in triplicate in final volume of 20µl, which included 5µl of cDNA or reference plasmid. Among Group I, Q-PCR detected BCR-ABL1 in 57 of the 118 patient samples assayed, with a median of 16.72 transcripts (range 1.38-47450). In only 2 (2.25 and 4.96 transcripts) of these 57 samples, ddPCR failed to detect BCR-ABL1 transcripts. Q-PCR and ddPCR failed to detect BCR-ABL1 in 45 (38.1%) samples. For 14 (11.8%) of the samples ddPCR was positive, median 3.4 copies (range 3-15) but negative by Q-PCR. Among Group II, one of the 21 samples was excluded from the analysis because <5500 droplets were generated. Of the 21 samples 7 were negative by ddPCR. Nested PCR was negative for all 7 samples. Three samples were positive by all 3 technologies, nested PCR, Q-PCR and ddPCR. Remarkably, 5 of the 21 samples were negative by nested, but positive by ddPCR; median 18.0 copies (6.2-22.0). These 5 samples were not subjected to Q-PCR. In Group III all 16 NC samples were negative by Q-PCR and ddPCR. In Group IV, ddPCR did detect BCR-ABL1 in the serially diluted IR sample calculated to have 1 copy (26 positive droplets of the 106095 total droplets), but Q-PCR failed. However, the lower dilutions, calculated to contain 0.1, 0.01 and 0.001 copies were negative by ddPCR and Q-PCR. We assayed 161 samples by ddPCR and Q-PCR, of these 139 were from CML patients. In addition 21 of the samples were also subjected to nested PCR. Our data support the notion ddPCR is at least one log more sensitive than Q-PCR. Of the 140 patient samples assayed, 19 (13.5%) were positive by ddPCR but negative by Q-PCR. Only 2 of the 118 samples in Group I were negative by ddPCR but positive by Q-PCR. There was in insufficient sample to repeat these 2 assays. The increased sensitivity of ddPCR as implied by the clinical samples was supported by assays performed using the IR. The serial dilution equivalent to 1.0 BCR-ABL1 copy was reliably detected by ddPCR, but was negative by Q-PCR. In summary, these data suggest ddPCR is more sensitive. However, the clinical significance of this must be assessed in context of long-term clinical outcome of patients with detectable BCR-ABL1 by ddPCR and negative by Q-PCR. But, clearly increased sensitivity is likely to enhance safe withdrawal of IM therapy for CML patients in CMR. Furthermore, regular monitoring of these patients by ddPCR would enable early detection of molecular relapse and thereby minimize the risk of disease progression. Disclosures Le Coutre: Novartis: Honoraria.

scholarly journals Pigment production by Cryptococcus neoformans and other Cryptococcus species from aminophenols and diaminobenzenes

1978 ◽  
Vol 7 (2) ◽  
pp. 146-152
Author(s):  
S Chaskes ◽  
R L Tyndall
Keyword(s):  
Cryptococcus Neoformans ◽  
Species Group ◽  
Pigment Production ◽  
The Other ◽  
Aminosalicylic Acid ◽  
Pigmentation Pattern ◽  
Unique Pattern ◽  
Group I ◽  
Cryptococcus Species ◽  
Group Ii

Cryptococcus neoformans and other Cryptococcus species can produce pigment(s) from many aminophenol and diaminobenzene compounds. Pigment production from these compounds is similar to the conversion of diphenols to melanin by C. neoformans. Several pigmentation patterns (resulting in the identification or grouping of Cryptococcus species) have been observed by using diaminobenzene and aminophenol compounds as substrates. The most common pigmentation pattern observed was pigment production by both C. neoformans and C. terreus. In contrast to the diphenols, only two aminophenols (4-hydroxymetanilamide and 3-aminotyrosine) were found to be highly specific as substrates. They allowed only C. neoformans to produce pigment. When 4-aminosalicylic acid was the substrate, a unique pattern was observed because only C. terreus, C. diffluens, and C. albidus produced pigment. Finally, a pattern was observed in which C. neoformans produced large amounts of pigment from aminophenol and diaminobenzene compounds, whereas the other Cryptococcus species produced smaller amounts. A simplified scheme with three substrates resulted in the identification of C. terreus and C. neoformans as well as two groups of other Cryptococcus species, group I (C. albidus and C. diffluens) and group II (C. laurentii and C. luteolus).

Phylogenetic analysis of the Pantomorus-Naupactus complex (Coleoptera: Curculionidae: Entiminae) from North and Central America

Zootaxa ◽  
2011 ◽  
Vol 2780 (1) ◽  
pp. 1 ◽  
Author(s):  
MARÍA V. ROSAS ◽  
JUAN J. MORRONE ◽  
M. GUADALUPE DEL RÍO ◽  
ANALÍA A. LANTERI
Keyword(s):  
Phylogenetic Analysis ◽  
Central America ◽  
Great Plains ◽  
Type Species ◽  
Cladistic Analysis ◽  
The United States ◽  
Species Group ◽  
South American Species ◽  
Group I ◽  
Group Ii

We undertook the first cladistic analysis of the Pantomorus-Naupactus complex (Coleoptera: Curculionidae) from North and Central America, based on 35 species and 61 morphological characters, plus 1151 bp of the mtDNA COI and Cyt b genes. The morphological and the combined matrices analyzed with TNT yielded a single most parsimonious cladogram that allows recognition of two main lineages within the Pantomorus-Naupactus complex in North and Central America. One is represented by the species formerly placed in Phacepholis and the Pantomorus species group II sensu Sharp, ranging along the Pacific coast of Central America and Mexico and reaching the Great Plains of North America in the United States, yet not occurring in South America. The other lineage is represented by the species of Naupactus and Pantomorus species group I, with closer relationships to the South American species of these genera. The Pantomorus group I includes the type species of the genus (P. albosignatus Boheman) whereas the Pantomorus group II includes the type species of Athetetes Pascoe, 1886 (A. globicollis Pascoe). Based on the results of our phylogenetic analysis, we recommend retaining the name Pantomorus Schoenherr for most species of the Pantomorus group I, except P. stupidus (Boheman) and P. femoratus Sharp which should be transferred to Naupactus Dejean. Moreover, we enlarge the previous concept of Phacepholis to include most species of the Pantomorus group II, and we establish the synonymy of Athetetes Pascoe, 1886 with Phacepholis Horn, 1876, being the latter the valid name, by priority.

Eulepida mbala, a new species from Zambia (Coleoptera: Scarabaeidae: Melolonthinae)

Zootaxa ◽  
2018 ◽  
Vol 4399 (4) ◽  
pp. 591
Author(s):  
RICHARD SEHNAL
Keyword(s):  
New Species ◽  
Type Species ◽  
Morphological Characters ◽  
Species Group ◽  
Undescribed Species ◽  
Group Iii ◽  
Group I ◽  
Group Ii ◽  
One New Species ◽  
A New Species

The genus Eulepida Kolbe, 1894 (Coleoptera: Scarabaeidae: Melolonthinae: Leucopholini) was established to accommodate 10 Afrotropical species, seven new and three previously placed in Lepidiota Kirby, 1828, Proagosternus Blanchard, 1851, and Tricholepis Hampson, 1891. Lacroix (2010) designated Leucopholis lepidota Klug, 1855 as the type species of the genus Eulepida. Currently the genus contains 20 species divided into three groups based on morphological characters (Lacroix 2010, 2013): species group I includes Eulepida lepidota (Klug, 1855), E. minor Moser, 1913, E. nitidicollis Kolbe, 1894, E. nyassica Kolbe, 1894, E. sinuatifrons (Fairmaire, 1887), and E. zambiensis Lacroix, 2010; species group II includes E. anatina Brenske, 1896, E. tschindeana Péringuey, 1904, and E. werneri Lacroix, 2010; and species group III includes E. baumanni Kolbe, 1894, E. flavovestita Moser, 1913, E. gracilipes Kolbe, 1894, E. kameruna (Frey, 1972), E. kenyensis Lacroix, 2010, E. mamboiae Brenske, 1896, E. manowensis Moser, 1913, E. mashona Arrow, 1902, E. montana Kolbe, 1894, E. reichei (Thomson, 1858), and E. savagei (Hope, 1842). Examination of material recently collected in Zambia revealed an undescribed species belonging to species group II (sensu Lacroix 2010). This group is defined by the combination of the following characters: protibia bidentate; antennal club distinctly longer than antennal shaft; pygidium narrow, longer than wide, with a pronounced elongate terminal invagination; and parameres symmetrical, long, evenly curved in ventral aspect (Lacroix 2010). The purpose of this paper is to describe one new species, to add new geographic records for some Eulepida species of group II, and to update the key for this group. New faunistic records are reported for Eulepida tschindeana and Eulepida werneri from Zimbabwe. 

scholarly journals Mornitoring and Identification of Human Astrovirus from Groundwater in Korea Based on Highly Sensitive RT-nested PCR Primer Sets

2021 ◽  
Vol 27 (4) ◽  
pp. 255-263
Author(s):  
Siwon Lee ◽  
Kyung Seon Bae ◽  
Jihyun Park ◽  
Jin-Ho Kim ◽  
Jin-Young Lee ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Highly Sensitive ◽  
Human Astrovirus ◽  
Primer Sets ◽  
Pcr Primer

Ultrastructural Lesions Produced by Alcohol and Sucrose in the Heart and Liver of C57BL Mice

1970 ◽  
Vol 28 ◽  
pp. 208-209
Author(s):  
K.K. SEKHRI ◽  
C.S. ALEXANDER ◽  
H.T. NAGASAWA
Keyword(s):  
Osmium Tetroxide ◽  
Male Mice ◽  
Alcohol Group ◽  
Group Iii ◽  
Group I ◽  
C57bl Mice ◽  
Group Ii

C57BL male mice (Jackson Lab., Bar Harbor, Maine) weighing about 18 gms were randomly divided into three groups: group I was fed sweetened liquid alcohol diet (modified Schenkl) in which 36% of the calories were derived from alcohol; group II was maintained on a similar diet but alcohol was isocalorically substituted by sucrose; group III was fed regular mouse chow ad lib for five months. Liver and heart tissues were fixed in 2.5% cacodylate buffered glutaraldehyde, post-fixed in 2% osmium tetroxide and embedded in Epon-araldite.

Heterogeneity and Immunochemical Properties of Anti-β2-glycoprotein I Autoantibodies

1998 ◽  
Vol 80 (09) ◽  
pp. 393-398 ◽  
Author(s):  
V. Regnault ◽  
E. Hachulla ◽  
L. Darnige ◽  
B. Roussel ◽  
J. C. Bensa ◽  
...  
Keyword(s):  
Fluid Phase ◽  
Anticardiolipin Antibodies ◽  
Dual Reactivity ◽  
Group Iii ◽  
Important Group ◽  
Epitope Specificity ◽  
Glycoprotein I ◽  
Group I ◽  
Group Ii ◽  
Sufficient Concentration

SummaryMost anticardiolipin antibodies (ACA) associated with antiphospholipid syndrome (APS) are directed against epitopes expressed on β2-glycoprotein I (β2GPI). Despite a good correlation between standard ACA assays and those using purified human β2GPI as the sole antigen, some sera from APS patients only react in the latter. This is indicative of heterogeneity in anti-β2GPI antibodies. To characterize their reactivity profiles, human and bovine β2GPI were immobilized on γ-irradiated plates (β2GPI-ELISA), plain polystyrene precoated with increasing cardiolipin concentrations (CL/β2GPI-ELISA), and affinity columns. Fluid-phase inhibition experiments were also carried out with both proteins. Of 56 selected sera, restricted recognition of bovine or human β2GPI occurred respectively in 10/29 IgA-positive and 9/22 IgM-positive samples, and most of the latter (8/9) were missed by the standard ACA assay, as expected from a previous study. Based on species specificity and ACA results, IgG-positive samples (53/56) were categorized into three groups: antibodies reactive to bovine β2GPI only (group I) or to bovine and human β2GPI, group II being ACA-negative, and group III being ACA-positive. The most important group, group III (n = 33) was characterized by (i) binding when β2GPI was immobilized on γ-irradiated polystyrene or cardiolipin at sufficient concentration (regardless of β2GPI density, as assessed using 125I-β2GPI); (ii) and low avidity binding to fluid-phase β2GPI (Kd in the range 10–5 M). In contrast, all six group II samples showed (i) ability to bind human and bovine β2GPI immobilized on non-irradiated plates; (ii) concentration-dependent blockade of binding by cardiolipin, suggesting epitope location in the vicinity of the phospholipid binding site on native β2GPI; (iii) and relative avidities approximately 100-fold higher than in group III. Group I patients were heterogeneous with respect to CL/β2GPI-ELISA and ACA results (6/14 scored negative), possibly reflecting antibody differences in terms of avidity and epitope specificity. Affinity fractionation of 23 sera showed the existence, in individual patients, of various combinations of antibody subsets solely reactive to human or bovine β2GPI, together with cross-species reactive subsets present in all samples with dual reactivity namely groups III and II, although the latter antibodies were poorly purified on either column. Therefore, the mode of presentation of β2GPI greatly influences its recognition by anti-β2GPI antibodies with marked inter-individual heterogeneity, in relation to ACA quantitation and, possibly, disease presentation and pathogenesis.

Treatment of venous leg ulcers with sulodexide

Phlebologie ◽  
2003 ◽  
Vol 32 (05) ◽  
pp. 115-120 ◽  
Author(s):  
A. Franek ◽  
H. Koziolek ◽  
M. Kucharzewski
Keyword(s):  
Pharmacological Treatment ◽  
Healing Process ◽  
Leg Ulcers ◽  
Venous Ulceration ◽  
Venous Leg Ulcers ◽  
Group I ◽  
Group Ii

SummaryAim: The study of the influence of sulodexide in the treatment of venous leg ulcers. Patients and method: 44 patients with chronic venous ulceration were randomly divided into two groups. Group I: 21 patients (ulceration area: 12.7-18.9 cm2), Group II: 23 patients (ulceration size: 12.1-20.3 cm2). Both groups were treated by using Unna’s boot. This dressing was changed every seven days until the ulcer had healed. Additionally, the patients in group II received the systemic pharmacological treatment with sulodexide. Results: After 7 weeks of treatment ulcers of seven patients (35%) from group I had healed, and 3 weeks later the ulceration of two more patients had healed completely. After further 7 weeks the ulcers of 12 patients had healed completely. Whereas in group II after 7 weeks of treatment ulceration of 16 (70%, p <0.05) patient had healed completely and after further 3 weeks the ulcers of the remaining 7 patients had healed, too. Conclusion: The use of sulodexide in patients with chronic venous leg ulcers accelerates the healing process.

Effects of Rest and Exercise on Cardiac Blood Volume Determinations

1997 ◽  
Vol 36 (08) ◽  
pp. 259-264
Author(s):  
N. Topuzović
Keyword(s):  
Radionuclide Ventriculography ◽  
Left Ventricular ◽  
Peak Exercise ◽  
Volume Determination ◽  
Group I ◽  
Lv Volumes ◽  
Group Ii ◽  
Lv Volume

Summary Aim: The purpose of this study was to investigate the changes in blood activity during rest, exercise and recovery, and to assess its influence on left ventricular (LV) volume determination using the count-based method requiring blood sampling. Methods: Forty-four patients underwent rest-stress radionuclide ventriculography; Tc-99m-human serum albumin was used in 13 patients (Group I), red blood cells was labeled using Tc-99m in 17 patients (Group II) in vivo, and in 14 patients (Group III) by modified in vivo/in vitro method. LV volumes were determined by a count-based method using corrected count rate in blood samples obtained during rest, peak exercise and after recovery. Results: In group I at stress, the blood activity decreased by 12.6 ± 5.4%, p <0.05, as compared to the rest level, and increased by 25.1 ± 6.4%, p <0.001, and 12.8 ± 4.5%, p <0.05, above the resting level in group II and III, respectively. This had profound effects on LV volume determinations if only one rest blood aliquot was used: during exercise, the LV volumes significantly decreased by 22.1 ± 9.6%, p <0.05, in group I, whereas in groups II and III it was significantly overestimated by 32.1 ± 10.3%, p <0.001, and 10.7 ± 6.4%, p <0.05, respectively. The changes in blood activity between stress and recovery were not significantly different for any of the groups. Conclusion: The use of only a single blood sample as volume aliquot at rest in rest-stress studies leads to erroneous estimation of cardiac volumes due to significant changes in blood radioactivity during exercise and recovery.

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