Phenotypic and Molecular Characterization of Staphylococcaceae and Enterobacteriaceae Species Isolated from Smoked, Dried, and Braised Fish Marketed in Ouagadougou

Background: To preserve its nutritional properties, fish must have good sanitary quality. The objective of this study was to investigate some pathogens contaminating smoked, dried, and braised fish marketed in Ouagadougou. Methodology: Potential pathogens of Enterobacteriaceae and Staphylococcus were screened in eight (8) species of processed fish. The investigation of the germs was carried out following the normative methods of microbiology. The identities of the strains were determined by API 20 E (BioMerieux S.A., France) and API STAPH (BioMerieux S.A., France) kits for Enterobacteriaceae and Staphylococcus species respectively. The uidA gene profile in Escherichia coli isolates was determined by simplex PCR. The identity of Staphylococcus aureus was confirmed by amplification of specific 23S rDNA regions and nuc gene profile with PCR. Results: A total of 235 fish samples were analyzed. A diversity of Enterobacteriaceae and Staphylococcus was detected. Twenty species of Enterobacteriaceae were identified among which, the most frequent were Escherichia coli, Salmonella sp, Raoultella ornithinolytica and Serratia odorifera, respectively in 22.6%, 4.3%, 28.9%, 17.4% of the samples analyzed. However, eleven species of Staphylococcus were identified among which, Staphylococcus xylosus, Staphylococcus aureus, Staphylococcus lugdunensis, and Staphylococcus sciuri were the most frequent with respective percentages of 47.7%, 23.4%, 12.8% and 10.6% of samples. For all the samples, the species frequently isolated were: Raoultella ornithinolytica, Escherichia coli, Serratia odorifera, Staphylococcus aureus, Staphylococcus xylosus, and Staphylococcus lugdunensis. The uidA gene specific to Escherichia coli was detected in 82.85% of strains (29/35). Amplification of the specific 23S rDNA region using staur primers was observed in 98% (49/50) of the isolated Staphylococcus aureus strains and the nuc gene was detected in 86% of Staphylococcus aureus strains. Conclusion: The isolated bacteria are potential pathogens involved in foodborne illnesses and intoxications. Effective sanitary safety systems must be implemented to guarantee the sanitary quality of fish supplied to consumers.

Molecular characterization of polar heteroatom species in oilsands bitumen-derived vacuum residue fractions by Fourier transform ion cyclotron resonance mass spectrometry

A Canadian in situ oilsands bitumen-derived vacuum residue(VR)was subjected to supercritical fluid extraction and fractionation(SFEF)into 13 extractable fractions and an unextractable end-cut and characterized by positive-and negative-ion electrospray ionization(ESI)Fourier transform ion cyclotron resonance mass spectrometry(FT-ICR MS).The results of negative-ion ESI FT-ICR MS showed that the N1 class species was the most abundant and the multifunctional group compounds,such as N1 O1,N1 O2,N1 S1,N1 S2,and N2 class species became abundant as the SFEF fraction became heavier.In positive-ion ESI mode,the relative abundance of N1 class species decreased gradually in the heavy SFEF fractions while that of multifunctional group compounds increased.The relative abundance of N4 V1 O1 increased dramatically in heavy fractions and the end-cut.The distributions of polar heteroatom species of VR derived from oilsands bitumen were similar with those of VR derived from the Venezuela Orinoco extra heavy oil.

Detection and Molecular Characterization of <i>Cronobacter sakazakii</i>Isolated from Powdered Infant Formula (PIF) from North Central Region, Nigeria

Cronobacter sakazakii is an emerging ubiquitous and opportunistic pathogen that currently contaminates a wide spectrum of foods including powdered milk and poses a lethal threat to neonates, the elderly and persons with immune deficiencies. They cause life threatening neonatal meningitis, septicemia, and necrotizing enterocolitis. A total of 360 samples of powdered infant formula were collected from postnatal hospital attendees reconstituting the PIF for their children in the North Central region of Nigeria where cases of infant mortality are very high and presenting as enterocolitis and diarrhea. Pre-enriched samples were cultured in chromogenic Cronobacter broth and were then further sub-cultured into a chromogenic Cronobacter sakazakii agar. They were positive, exhibiting yellowish cultures typical of Cronobacter sakazakii. Biochemical tests of the isolates were also carried out and indicated the presence of Cronobacter sakazakii. The isolates were then characterized molecularly using specie specific PCR detection of Cronobacter sakazakii. The targeted genes of interest were ompA gene and CPA gene. The isolates tested showed bands for ompA gene on electrophoresis imager and were confirmed as Cronobacter sakazakii. In Nigeria, majority of infants are still fed with PIF. There is no existing data on the detection of Cronobacter sakazakii previously reported in the North central region of Nigeria hence the need to carry out the present study. The result of the study demonstrated the need for effective prevention and control measures as contamination of PIF with Cronobacter sakazakii constituted potential public health risk to neonates and infants.

Preliminary Phenotypic and SNP-Based Molecular Characterization of Maize (<i>Zea mays</i>L.)-Mexicana (<i>Zea mays</i>SSP. <i>Mexicana</i>) Introgression Lines under Inbred Background of 48-2

Wild relatives possess potential genetic diversity for maize (Zea mays L.) improvement. Characterization of maize-mexicana introgression lines (ILs) is of great value to diversify the genetic base and improve the maize germplasm. Four maize-mexicana IL generations, i.e. BC1, BC2, BC3, and RIL, were constructed under the elite inbred background of 48-2, elite inbred line that is widely used in maize breeding in Southwestern China, and were phenotyped in different years and genotyped with 56110 SNPs. The results indicated that 48-2 had higher phenotypic performances than all the characterized ILs on most of the agronomic traits. Compared with other ILs, BC2 individuals exhibited more similar performance to 48-2 on most traits and possessed the highest kernel ratio (66.5%). Population structure and principal component analysis indicated that BC3 individuals gathered closer to 48-2 and exhibited the lowest mexicana-introgression frequency (0.50%), while BC2 (29.06%) and RIL (18.52%) showed higher introgression frequency. The high level of genetic diversity observed in the maize-mexicana ILs demonstrated that Z. mays ssp. mexicana can serve as a potential source for the enrichment of maize germplasm.

Biochemical Liver Functions and Molecular Identification of Fasciola hepatica from Experimentally Infected Rat Model

The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2nd, 4th, 6th, 8th, 10th, 12th and 14th weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14th WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.

Origin and molecular characterization of the human-infecting H6N1 infl uenza virus in Taiwan

In June 2013,the fi rst human H6N1 infl uenza virus infec-tion was confirmed in Taiwan.However,the origin and molecular characterization of this virus,A/Taiwan/2/2013(H6N1),have not been well studied thus far.In the present report,we performed phylogenetic and coalescent analy-ses of this virus and compared its molecular profi le/char-acteristics with other closely related strains.Molecular characterization of H6N1 revealed that it is a typical avian infl uenza virus of low pathogenicity,which might not rep-licate and propagate well in the upper airway in mammals.Phylogenetic analysis revealed that the virus clusters with A/chicken/Taiwan/A2837/2013(H6N1)in seven genes,except PB1.For the PB1 gene,A/Taiwan/2/2013 was clus-tered with a different H6N1 lineage from A/chicken/Taiwan/A2837/2013.Although a previous study demonstrated that the PB2,PA,and M genes of A/Taiwan/2/2013 might be derived from the H5N2 viruses,coalescent analyses revealed that these H5N2 viruses were derived from more recent strains than that of the ancestor of A/Taiwan/2/2013.Therefore,we propose that A/Taiwan/2/2013 is a reassor-tant from different H6N1 lineages circulating in chickens in Taiwan.Furthermore,compared to avian isolates,a sin-gle P186L(H3 numbering)substitution in the hemaggluti-nin H6 of the human isolate might increase the mammali-an receptor binding and,hence,this strain’s pathogenicity in humans.Overall,human infection with this virus seems an accidental event and is unlikely to cause an infl uenza pandemic.However,its co-circulation and potential reas-sortment with other infl uenza subtypes are still worthy of attention.

Molecular Screening of Rice Cultivated in Benin for the Identification of Xanthomonas oryzae Pv. oryzae and Bacterial Leaf Blight Resistance Genes

One of the most devastating diseases of rice worldwide is bacterial blight (BLB) caused by Xanthomonas oryzae pv. Oryzae (Xoo). In Benin, Xoo was first described in 2013 on wild rice Oryzae longistaminata. So far, no study has been done on Beninese Xoo strains. We do not know whether the pathogen has already passed into the rice varieties grown, or if they are exposed to other bacteria. Whereas the use of resistant varieties, carrying resistance genes, is the only highly effective and environmentally friendly way to control this disease, no information is available on these Xoo resistance genes in rice varieties grown in Benin apart from the one we recently. This study aims to identify Beninese Xoo strains, causing BLB and screen rice varieties grown in Benin for the main resistance genes. Diseased rice leaves showing typical symptoms of fire blight collected from different rice fields in the three phytogeographic areas of Benin were analyzed by PCR for Xoo-specific sequence identification. Furthermore, seventy-five collected rice accessions were screened to identify xa5, Xa7, xa13, and Xa21 resistance genes to Xoo. The results reveal that Xanthomonas oryzae was identified in two fields in Banikouara and one in Malanville. On the other hand, Sphingomonas sp. has been identified in several other rice fields in Benin. Forty-seven of seventy-five rice accessions examined (62.66%) carried Xoo resistance genes with 3 (4%) and 40 (53.33%) of xa5 and Xa21 respectively. None of the accessions had either Xa7 or xa13 resistance genes. Three accessions possess both xa5 and Xa21 genes. Isogenic lines IRBB60 and IRBB21, supposed to be a positive control, presented a Xoo sensitivity allele. These results indicate that Xoo has moved from the wild rice variety to the cultivated variety in northern Benin and varietal improvement programs must be implemented with varieties having several resistance genes for the efficient response against a possible BLB pandemic in Benin.

Toward Molecular Cytogenetical Characterizations in Cotton Genome

Cotton is viewed as the most important cash crop in the world,and sustains the agricultural economies of many nations by providing a sustainable fiber product for the textile industry.Due to

Molecular Cloning and Characterization of the Actin-depolymerizing Factor Gene in Gossypium barbadense

Sea Island cotton(Gossypium barbadense L.) has been highly valued in Verticillium wilt resistance and many fiber qualities including fiber length,strength,and fineness.To identify whether

Molecular Cloning and Characterization of Genes Involved in Cotton(Gossypium barbadense L.) Response to Verticillium dahliae

Verticillium dahliae Kleb.is a necrotrophic plant pathogen which causes serious soil borne vascular disease in cotton.The molecular basis the defense response of cotton to this pathogen is

Molecular Weight Characterization of Partially Hydrolyzed Polyacrylamide in the Complex Salts Solution

The effects of the nature of salts, salt concentration, pH value, degree of hydrolysis as well as surfactant on molecular size of HPAM were studied- It is found that [q] of HPAM changes with all the above variables, and the molecular weigh ofHPAM can be obtained through [q] measurement at a specific salts environment in which the [q] is independent of above factors.

Molecular Cloning,Expression,and Characterization of an Adenylyl Cyclase-associated Protein from Gossypium arboreum Fuzzless Mutant

CAP,an adenylyl cyclase-associated protein,is predicted to be involved in cytoskeletal organization and signal transduction.Recently,we found that CAP may play an important role in fuzz-like fiber cell initiation in cotton.For the further research,we isolated two CAP homologues from wild

Prevalence and methodologies for detection,characterization and subtyping of Listeria monocytogenes and L.ivanovii in foods and environmental sources

Listeria monocytogenes,one of the most important foodborne pathogens,can cause listeriosis,a lethal disease for humans.L.ivanovii,which is closely related to L.monocytogenes,is also widely distributed in nature and infects mainly warm-blooded ruminants,causing economic loss.Thus,there are high priority needs for methodologies for rapid,specific,cost-effective and accurate detection,characterization and subtyping of L.monocytogenes and L.ivanovii in foods and environmental sources.In this review,we(A)described L.monocytogenes and L.ivanovii,world-wide incidence of listeriosis,and prevalence of various L.monocytogenes strains in food and environmental sources;(B)comprehensively reviewed different types of traditional and newly developed methodologies,including culture-based,antigen/antibody-based,LOOP-mediated isothermal amplification,matrix-assisted laser desorption ionization-time of flight-mass spectrometry,DNA microarray,and genomic sequencing for detection and characterization of L.monocytogenes in foods and environmental sources;(C)comprehensively summarized different subtyping methodologies,including pulsed-field gel electrophoresis,multi-locus sequence typing,ribotyping,and phage-typing,and whole genomic sequencing etc.for subtyping of L.monocytogenes strains from food and environmental sources;and(D)described the applications of these methodologies in detection and subtyping of L.monocytogenes in foods and food processing facilities.

A Review of Prevalence, Antimicrobial Susceptibility Patterns and Molecular Characteristics of Methicillin-Resistant Staphylococcus aureus (MRSA) in the Caribbean

Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen causing infections resulting in severe morbidity and mortality worldwide. To date, the true nature and extent of MRSA infections in the Caribbean are not well understood. This is a review of the limited studies in the Caribbean describing the prevalence, epidemiology, and molecular characteristics of MRSA in hospitalized and non-hospitalized patients. Relevant articles were searched and extracted from PubMed and Mendeley and a narrative review of the findings was constructed. An aggregate of 24 articles, from 1999 to 2020, was found from 10 of 27 countries. Majority of the studies were from Trinidad and Tobago (29%) and Jamaica (21%) while 50% were from Barbados, Dominican Republic, Martinique, Haiti, Cuba, St. Kits & Nevis, Guadeloupe, and Guyana. Approximately 75% of investigations were conducted on hospitalized patients versus 20% on outpatients. The data revealed geographical differences in the prevalence of MRSA within the Caribbean;20% - 100% of Staphylococcus aureus clinical isolates from hospitalized patients and outpatients were resistant to methicillin, macrolides, and fluoroquinolones, but susceptible to several non-beta lactam antibiotics, due to the widespread occurrence of CA-MRSA clone ST8 SCCmec IV, PVL positive. There was moderate prevalence of ST72 SCCmec V (14% - 25%) in both hospital and community settings in a few of the countries while ST30 SCCmec IV, PVL positive, was moderately prevalent (27%) only in Dominican Republic. Also, there was moderate prevalence of HA-MRSA ST5 SCCmec II (18%) in community settings in the Dominican Republic and Martinique, but high prevalence of HA-MRSA ST239 SCCmec III (60%) in hospitalized patients in Cuba and Trinidad & Tobago. The epidemiologic profile of MRSA in both hospital and community settings is changing in the Caribbean. Epidemiological studies on outpatient settings and the implementation of stringent hospital infection control measures are needed in the region.

Helicobacter pylori CagA protein polymorphisms and their lack of association with pathogenesis

AIM: To investigate Helicobacter pylori (H. pylori) CagA diversity and to evaluate the association between protein polymorphisms and the occurrence of gastric pathologies. METHODS: One hundred and twenty-two clinical isolates of H. pylori cultured from gastric biopsies obtained from Colombian patients with dyspepsia were included as study material. DNA extracted from isolates was used to determine cagA status, amplifying the C-terminal cagA gene region by polymerase chain reaction. One hundred and six strains with a single amplicon were sequenced and results were used to characterize the 3' variable region of the cagA gene. To establish the number and type of tyrosine phosphorylation motifs Glutamine acid-Proline-Isoleucine-Tyrosine-Alanine (EPI-YA) bioinformatic analysis using Amino Acid Sequence Analyzer-Amino Acid Sequence Analyzer software was conducted. Analysis of the association between the number of EPIYA motifs and the gastric pathology was performed using χ2 test and analysis of the presence of EPIYA-C motifs in relation to the pathology was made by logistic regression odds ratios. Comparisons among EPIYA types found and those reported in GenBank were performed using a proportion test in Statistix Analytical Software version 8.0. RESULTS: After amplification of the 3' of the cagA gene, 106 from 122 isolates presented a single amplicon and 16 showed multiple amplicons. As expected, diversity in the size of the cagA unique fragments among isolates was observed. The 106 strains that presented a single amplicon after 3' cagA amplification came from patients with gastritis (19 patients), atrophic gastritis (21), intestinal metaplasia (26), duodenal ulcer (22) and gastric cancer. DNA sequence analysis showed that the differences in size of 3' cagA unique fragments was attributable to the number of EPIYA motifs: 1.9% had two EPIYA motifs, 62.3% had three, 33.0% had four and 2.8% had five motifs. The majority of tested clinical strains (62.3%) were found to harbor the ABC combination of EPIYA motifs and a significant statistical difference was observed between the frequencies of ABCC tyrosine phosphorylation motifs and Western strains sequences deposited in GenBank. CONCLUSION: The present report describes a lack of association between H. pylori CagA-protein polymorphisms and pathogenesis. ABCC high frequency variations compared with Western-strains sequences deposited in GenBank require more investigation.

Distribution of nitrogen and oxygen compounds in shale oil distillates and their catalytic cracking performance

The positive-and negative-ion electrospray ionization(ESI)coupled with Fourier transform-ion cyclotron resonance mass spectrometry(FT-ICR MS)was employed to identify the chemical composition of heteroatomic compounds in four distillates of Fushun shale oil,and their catalytic cracking performance was investigated.There are nine classes of basic nitrogen compounds(BNCs)and eleven classes of non-basic heteroatomic compounds(NBHCs)in the different distillates.The dominant BNCs are mainly basic N1 class species.The dominant NBHCs are mainly acidic O2 and O1 class species in the300-350℃,350-400℃,and 400-450℃distillates,while the neutral N1,N1 O1 and N2 compounds become relatively abundant in the>450℃fraction.The basic N1 compounds and acidic O1 and O2 compounds are separated into different distillates by the degree of alkylation(different carbon number)but not by aromaticity(different double-bond equivalent values).The basic N1 O1 and N2 class species and neutral N1 and N2 class species are separated into different distillates by the degrees of both alkylation and aromaticity.After the catalytic cracking of Fushun shale oil,the classes of BNCs in the liquid products remain unchanged,while the classes and relative abundances of NBHCs vary significantly.

The journey of personalizing gastric cancer treatment

Gastric cancer ranks the fourth most prevalent malignancy yet it is the second leading cause of cancer-related death.Every year,gastric cancer adds nearly 1 million new cancer cases,and 723,000 or 10% of cancer deaths to the global cancer burden.Approximately,405,000 or 43% of the new cases and 325,000 or 45% of the deaths are in China,making gastric cancer a particularly challenging malignancy.This thematic series discusses the molecular classifications of gastric cancer by the Cancer Genome Atlas(TCGA) and the Asian Cancer Research Group(ACRG) as well as the implications in personalized therapeutic choices;discusses the evolution of gastric surgery and presents perspectives on surgical techniques in treating gastric cancer;and reviews current and emerging targeted agents as well as immunotherapies in treating gastric cancer.With these advancements in molecular characterization,surgical intervention,and targeted and immunotherapies,gastric cancer will enter a personalized medicine era in the next 5 years.

Mayaro fever:A brief review on the immune profile

Mayaro virus is an emergent alphavirus that infects humans,leading to Mayaro fever.Approximately fifty percent of infected patients develop arthritis symptoms in the recovery phase,a phase that can last up to a year.The literature about Mayaro virus infection and its immune response is scarce,which may hamper the development of treatment strategies.We summarize changes in cytokines and chemokines in the acute and recovery phase in Mayaro virus infected patients,and relate this molecular characterization with the immune response.VEGF and IL-12/p70 show pronounced changes in patients in the acute phase,suggesting the development of cellular immunity and Th1 response.IL-6,IL-7,CXCL8/IL-8,IL-13,IL-17,and IFN-γare elevated in patients with arthritis symptoms in the long-term recovery phase,which may be related to the continuous inflammatory process,a possible Th2 inhibiting and promoting Th17 process.Although few studies discuss the issue,with a small number of patients and different backgrounds,inflammatory and immune response and manifestations seem to be closely linked.This information may help to develop the appropriate treatment strategies in Mayaro virus infection.Therefore,we analyzed and summarized data available in literature.

Comparison of RF3 <i>B. juncea</i>to RF3 <i>B. napus</i>

RF3 Brassica napus L. (InVigorTM Canola) has been in commercial cultivation since 1997. RF3 B. napus was produced by Agrobacterium-mediated transformation to provide glufosniate ammonium herbicide tolerance to the crop. Canola quality (CQ) varieties of Brassica juncea L. (Czern & Coss), a close relative of B. napus, have been available in Canada for more than 20 years. Currently, BASF Agricultural Solutions is developing CQ RF3 B. juncea via conventional breeding with RF3 B. napus to provide similar herbicide tolerance in the B. juncea species. The insertion event RF3 contains the bar gene (origin Streptomyces hygroscopicus) coding for phosphinothricin acetyl transferase (PAT/bar) protein which confers tolerance to glufosinate-ammonium herbicides. RF3 also contains the barstar gene (origin Bacillus amyloliquefaciens), coding for the Barstar protein, which is an inhibitor of the Barnase protein. In the absence of Barnase, there is no impact of Barstar. Many safety studies have been conducted during the development of the canola quality lines of RF3 B. napus and RF3 B. juncea. This report seeks to compare the two species based on the scientific data generated during the safety assessments of RF3 B. napus and RF3 B. juncea and discusses the utility of using studies with one species to support the safety of the other species both containing the same insertion event.