Characterization of Lactic Acid Bacteria and Pathogens Isolated from Traditionally Fermented Foods, In Relation to Food Safety and Antimicrobial Resistance in Tribal Hill Areas of Northeast India

Traditional fermented food products are often connected to various indigenous tribes and thus vary due to ethnicity, geography, and natural resource availability. The indigenous tribes from India greatly rely on fermentation processes for food preservation, flavor, and nutrition. Fermented foods can provide health benefits but also pose risks from harmful microbes and contaminants that grow in the food due to poor hygiene. In this study, we identified lactic acid bacteria (LAB) in fermented food collected from Northeast India, assessed their beneficial properties, and highlighted the risk from food pathogens that have antimicrobial resistance traits. A total of 113 different samples of fermented food products were collected from the local markets of five Northeastern Indian states (Nagaland, Manipur, Meghalaya, Arunachal Pradesh, and Sikkim). Standard laboratory methods were used to isolate LAB and determine their probiotic properties, conduct coliform counts, and isolate presumptive staphylococci from the fermented food samples. Antimicrobial susceptibility was determined by using the BD-Phoenix 100 automated system. We isolated 30 LAB with probiotic potential. The average aerobic colony count in different fermented food was 4.4-7.7 log center dot cfu/g, while coliforms were present in 43% (49/113, 95% (CI 34-53)) of the food samples, indicating low-hygiene levels. Additionally, some food samples contained staphylococci with phenotypic antibiotic-resistance markers (MRS, HLMUP, BLACT, and STAMLS). This study indicates that probiotic bacteria could be present in traditional fermented food products of Northeast India, but contamination with staphylococci and other bacterial pathogens with antibiotic resistance traits could put the health of consumers at risk

Avian strains of emerging pathogen Escherichia fergusonii are phylogenetically diverse and harbor the greatest AMR dissemination potential among different sources: Comparative genomic evidence

IntroductionEscherichia fergusonii is regarded as an emerging pathogen with zoonotic potential. In the current study, we undertook source-wise comparative genomic analyses (resistome, virulome, mobilome and pangenome) to understand the antimicrobial resistance, virulence, mobile genetic elements and phylogenetic diversity of E. fergusonii.MethodsSix E. fergusonii strains (5 multidrug resistant strains and 1 biofilm former) were isolated from poultry (duck faeces and retail chicken samples). Following confirmation by phenotypic and molecular methods, the isolates were further characterized and their genomes were sequenced. Comparative resisto-virulo-mobilome analyses and pangenomics were performed for E. fergusonii genomes, while including 125 other E. fergusonii genomes available from NCBI database.Results and discussionAvian and porcine strains of E. fergusonii were found to carry significantly higher number of antimicrobial resistance genes (p < 0.05) and mobile genetic elements (plasmids, transposons and integrons) (p < 0.05), while the pathogenic potential of bovine strains was significantly higher compared to other strains (p < 0.05). Pan-genome development trends indicated open pan-genome for all strains (0 < γ < 1). Genomic diversity of avian strains was found to be greater than that from other sources. Phylogenetic analysis revealed close clustering among isolates of similar isolation source and geographical location. Indian isolates of E. fergusonii clustered closely with those from Chinese and a singleton Australian isolate. Overall, being the first pangenomic study on E. fergusonii, our analysis provided important cues on genomic features of the emerging pathogen E. fergusonii while highlighting the potential role of avian strains in dissemination of AMR

Incidence and virulence properties of E. coli isolated from fresh fish and ready-to-eat fish products

Aim: To investigate the incidence and virulence properties of E. coli in fresh fish and ready-to-eat fish products from retail markets of the Ludhiana the present study was conducted. Materials and Methods: Total of 184 samples comprising 96 raw fish and 88 ready- to-eat (RTE) fish products were collected from Ludhiana and other parts of Punjab and were subjected to suitable microbiological methods for E. coli isolation. E. coli isolates were subjected for haemolytic activity and indicators of plausible cytotoxicity (lecithinase, protease and gelatinase production), congo red dye biding assay. To assess virulence potential isolates were molecularly screened for stx 1 and 2 genes. Results: From raw fish samples 47(48.95%), E. coli, were isolated. From RTE fish products 7(12.96%), E. coli were isolated. Overall incidence for E. coli was 54 (29.34%). In vitro virulence characterization of isolates exhibited that all E. coli isolates were haemolytic while indicators of plausible cytotoxicity ( lecithinase, protease and gelatinase production) were in the range of 16.67% to 35.19% indicated that though the isolates were haemolytic they were perhaps less likely to be cytotoxic. Congo Red binding assay for E. coli isolates revealed that majority (88.89%) of the isolates failed to uptake the dye and only few (11.11%) could bind the dye. Results of serotyping revealed a total of 15 different serotypes among the E. coli isolates. More variation was observed among isolates from raw fish samples (12 serotypes) while RTE fish products harboured only 5 different serotypes. Molecular characterization of E. coli isolates revealed that PCR screening of isolates revealed that total 39 (72.22%) samples out of 54 E. coli isolates were positive for stx1 gene and 28 (51.85%) of isolates were positive for stx2 gene. Sources wise, 36 (66.66%) of isolates from raw fish and 3(5.55%) of isolates from RTE fish products were positive for stx1 while and stx2 gene could be detected in 24(44.44%) isolates from raw fish and 4(7.4%) isolates from RTE fish products.Interestingly, about 20% (37.03%) isolates were positive for both stx1 and stx2 genes. Among these multivirulent isolates majority (n=18) belonged to raw fish samples compared to a few (n=2) from RTE fish products. Conclusion: The results of the present study highlighted the possible risks to consumers of fish and fish products in the region that demand action to address this public health concern. [Vet World 2013; 6(1.000): 5-9

Postoperative acute anisocoria and old traumatic brain injury

Anisocoria is an uncommon entity in general postoperative intensive care. We present a case of a 45-year-old man suffering from severe acute pancreatitis with a past history of traumatic brain injury (TBI), who developed hypertension, bradycardia and anisocoria soon after re-exploration surgery under general anaesthesia. Computed tomography showed no new lesion. Measures directed towards reducing intracranial pressure resulted in amelioration in about 12h. The possible role of old TBI in the causation of anisocoria during general anaesthesia and resuscitation has been explored in this report

Whole genome sequencing and analysis of Campylobacter coli YH502 from retail chicken reveals a plasmid-borne type VI secretion system

Campylobacter is a major cause of foodborne illnesses worldwide. Campylobacter infections, commonly caused by ingestion of undercooked poultry and meat products, can lead to gastroenteritis and chronic reactive arthritis in humans. Whole genome sequencing (WGS) is a powerful technology that provides comprehensive genetic information about bacteria and is increasingly being applied to study foodborne pathogens: e.g., evolution, epidemiology/outbreak investigation, and detection. Herein we report the complete genome sequence of Campylobacter coli strain YH502 isolated from retail chicken in the United States. WGS, de novo assembly, and annotation of the genome revealed a chromosome of 1,718,974 bp and a mega-plasmid (pCOS502) of 125,964 bp. GC content of the genome was 31.2% with 1931 coding sequences and 53 non-coding RNAs. Multiple virulence factors including a plasmid-borne type VI secretion system and antimicrobial resistance genes (beta-lactams, fluoroquinolones, and aminoglycoside) were found. The presence of T6SS in a mobile genetic element (plasmid) suggests plausible horizontal transfer of these virulence genes to other organisms. The C. coli YH502 genome also harbors CRISPR sequences and associated proteins. Phylogenetic analysis based on average nucleotide identity and single nucleotide polymorphisms identified closely related C. coli genomes available in the NCBI database. Taken together, the analyzed genomic data of this potentially virulent strain of C. coli will facilitate further understanding of this important foodborne pathogen most likely leading to better control strategies. The chromosome and plasmid sequences of C. coli YH502 have been deposited in GenBank under the accession numbers CP018900.1 and CP018901.1, respectively

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Novel APC gene mutations associated with protein alteration in diffuse type gastric cancer

Abstract Background The role of adenomatous polyposis coli (APC) gene in mitosis might be critical for regulation of genomic stability and chromosome segregation. APC gene mutations have been associated to have a role in colon cancer and since gastric and colon tumors share some common genetic lesions, it is relevant to investigate the role of APC tumor suppressor gene in gastric cancer. Methods We investigated for somatic mutations in the Exons 14 and 15 of APC gene from 40 diffuse type gastric cancersamples. Rabbit polyclonal anti-APC antibody was used, which detects the wild-type APC protein and was recommended for detection of the respective protein in human tissues. Cell cycle analysis was done from tumor and adjacent normal tissue. Results APC immunoreactivity showed positive expression of the protein in stages I, II, III and negative expression in Stages III and IV. Two novel deleterious variations (g.127576C > A, g.127583C > T) in exon 14 sequence were found to generate stop codon (Y622* and Q625*)in the tumor samples. Due to the generation of stop codon, the APC protein might be truncated and all the regulatory features could be lost which has led to the down-regulation of protein expression. Our results indicate that aneuploidy might occurdue to the codon 622 and 625 APC-driven gastric tumorigenesis, in agreement with our cell cycle analysis. The APC gene function in mitosis and chromosomal stability might be lost and G1 might be arrested with high quantity of DNA in the S phase. Six missense somatic mutations in tumor samples were detected in exon 15 A-B, twoof which showed pathological and disease causing effects based on SIFT, Polyphen2 and SNPs & GO score and were not previously reported in the literature or the public mutation databases. Conclusion The two novel pathological somatic mutations (g.127576C > A, g.127583C > T) in exon 14 might be altering the protein expression leading to development of gastric cancer in the study population. Our study showed that mutations in the APC gene alter the protein expression and cell cycle regulation in diffuse type gastric adenocarcinoma

Polymerase Chain Reaction (PCR) Assay for Rapid Diagnosis and Its Role in Prevention of Human Brucellosis in Punjab, India

Objectives: Brucellosis is the most common zoonotic disease that has been diagnosed mainly by serological tests and blood culture to some extent. This study was designed to establish a PCR technique for rapid diagnosis to be used in surveillance activities. Methods: The purpose of this study was firstly explained to the study population and verbal consent was obtained before sample collection. Peripheral blood was collected from 116 occupationally exposed groups with and without pyrexia of unknown origin from various districts of Punjab. Samples were subjected to blood culture, serological tests and DNA extraction was done using conventional laboratory extraction procedure. A primer pair B4/B5 that amplifies a gene encoding a 31 kDa immunogenic outer membrane protein (bcsp31) of Brucella species was used for PCR amplification. Results: The results showed that 8 (7%) of the cases had positive PCR and the detection threshold of primers used in this study were 715 cfu/ml. PCR results were 51.3% accurate for sensitivity of 12.6% and specificity of 100% using STAT as gold standard. Conclusions: Early-case reporting is possible by rapid tests like PCR. Thus, PCR is a promising diagnostic tool for routine investigation and surveillance of brucellosis which is the key element for management of prevention and control programmes. But patient condition before testing, optimal clinical specimen, sample volume used, simple and efficient DNA extraction protocol are the points of concern for PCR to be used as a routine test in clinical laboratory practice

Whole-genome sequence data and analysis of a Staphylococcus aureus strain SJTUF_J27 isolated from seaweed

The complete genome sequence data of S. aureus SJTUF_J27 isolated from seaweed in China is reported here. The size of the genome is 2.8 Mbp with 32.9% G + C content, consisting of 2614 coding sequences and 77 RNAs. A number of virulence factors, including antimicrobial resistance genes (fluoroquinolone, beta-lactams, fosfomycin, mupirocin, trimethoprim, and aminocoumarin) and the egc enterotoxin cluster, were found in the genome. In addition, the genes encoding metal-binding proteins and associated heavy metal resistance were identified. Phylogenetic data analysis, based upon genome-wide single nucleotide polymorphisms (SNPs), and comparative genomic evaluation with BLAST Ring Image Generator (BRIG) were performed for SJTUF_J27 and four S. aureus strains isolated from food. The completed genome data was deposited in NCBI׳s GenBank under the accession number CP019117, https://www.ncbi.nlm.nih.gov/nuccore/CP019117. Keywords: Staphylococcus aureus, Genome assembly, Whole genome sequencing (WGS), Virulence facto