Salmonella Serotypes Research Articles

Salmonella meningitis with hydrocephalus as complication in children: A case series

Background: Bacterial meningitis by salmonella serotype is a rare infection among neonates and young children. The treatment of salmonella meningitis requires bactericidal agents that are capable of penetrating into cerebrospinal fluid (CSF). Acute complications of bacterial meningitis mainly include hydrocephalus, subdural collection, cerebral infarction, ventriculitis, empyema, intracranial abscess and cranial nerve palsy. Two Cases Presentation: First case, 1 months-old boy brought to the general hospital with chief complaint of fever. Patient had decreased of consciousness and bulging of anterior fontanella. Analysis of cerebrospinal fluid (CSF) showed Pandy test (+4) and None test (+4), high protein 369 mg/dl (mononuclear 44%, polymorphonuclear 56%), liquor glucose 2 mg/dl, CSF culture and blood culture resulted salmonella. Electrolyte and blood sugar was normal, complete blood count showed leukocytes 13.96x 103/µL and mild normochromic normocytic anemia. Head CT scan revealed communicating hydrocephalus. Second case, 6 months-old boy brought to general hospital with seizure as chief complaint. Cerebrospinal fluid (CSF) analysis showed Pandy test (+4) and None test (+4), high protein 569 mg/dL, liquor glucose 26 mg/dL, CSF culture and blood culture results found salmonella sp. Head CT scan revealed subdural hygroma at frontoparietal. Patient was treated by 3rd generation of cephalosporins (ceftriaxone) for 4 weeks. Patient discharge from hospital. Summary: Management of salmonella meningitis include administration of empiric antibiotic, supportive treatment and management of complication. High dose of antibiotic is given empirically without waiting for the results of culture. Learning Points: Clinical manifestations and outcome of meningitis salmonella

Salmonellosis in the COVID­19 Pandemic Era

Relevance. In 2020, there was a unique situation caused by the COVID-19 pandemic and the anti-epidemic measures introduced in this regard. To date, the question of how these methods affect the spread of other infectious diseases, including salmonellosis, has not been studied.Target. To assess the impact of anti-epidemic measures during the COVID-19 pandemic on the epidemic process of salmonella infection in St. Petersburg.Materials and methods. Reporting form No. 2 of Rospotrebnadzor «Information on infectious and parasitic diseases» for 2018–2020 and data from the State report «On the state of sanitary and epidemiological well-being of the population in St. Petersburg in 2019», «On the state of sanitary and epidemiological well-being of the population in St. Petersburg in 2020». Data processing was carried out using Microsoft Excel (Microsoft®, USA), Statistica for Windows (StatSoft®, USA) with the determination of the level of significance (p), the calculation of confidence intervals (95% CI) was carried out in the EpiTools application.Results. The incidence of salmonellosis in 2020 was significantly lower than in the previous 2 years 28.86 (95% CI 27.45–30.34) per 100 ths population, and in 2019 and 2018 – 49.8 (95% CI 47.9–51.7) and 39.21 (95% CI 37.6– 40.9) per 100 ths of the population respectively. When analyzing the incidence of salmonellosis in 1995 to 2020, it was revealed that in 2020 the incidence rate was the lowest over the past 25 years and below the multi-year average (37 per 100 ths population) in St. Petersburg by 22%. A decrease in the incidence of salmonellosis was observed in 2020 and in the Russian Federation as a whole compared to 2019, the indicator increased 1.6 times and amounted to 14.71 per 100 ths rubles. population (multi-year average – 29.1). In 2020, there was a change in the intra-annual incidence of salmonellosis. While the peak incidence of salmonellosis persisted in the autumn period (September–October), in 2020 there was no characteristic rise in the spring period, as was observed in 2018 and 2019, which may be due to the spring lockdown (p = 0.03). The decrease in the incidence of salmonellosis was due to a significant decrease in the incidence among adults, while this was not observed in other age groups. In 2018–2020, diseases in the population were caused by 61 serotypes of Salmonella: in 2018 – 33, in 2019 – 32, and in 2020 – 39 serotypes. During the pandemic, the decrease in the incidence of salmonellosis was due to the decrease in the incidence caused by S. Enteritis, which in 2020 amounted to 19.91 (95% CI 18.73-21.14) per 100 thousand population, while in 2019 – 39, 01 (95% CI 37.4–40.7) and in 2018 – 31.24 (95% CI 29.8–32.8) per 100 ths population. At the same time, the decrease in the incidence rate was a decrease in the incidence rate among adults, while in other age groups no changes in the incidence rate were observed. The incidence due to other salmonella serotypes did not change. There were no significant differences in the incidence of salmonellosis by sex and age.Conclusion. The measures introduced during the pandemic in 2020 led to a decrease in the incidence of salmonellosis in St. Petersburg by reducing the incidence of salmonellosis caused by S. Enteritidis among adults.

Characterization of resistance genes and plasmids from sick children caused by Salmonella enterica resistance to azithromycin in Shenzhen, China.

Samonella is 1 of 4 key global causes of diarrhoeal diseases, sometimes it can be serious, especially for yong children. Due to the extensive resistance of salmonella serotypes to conventional first-line drugs, macrolides (such as azithromycin) have been designated as the most important antibiotics for the treatment of salmonella. Antimicrobial resistance is a major public health problem in the world, and the mechanism of azithromycin resistance is rarely studied. This study determined the azithromycin resistance and plasmids of Salmonella enterica isolates from children attending the Shenzhen Children's Hospital. The susceptibility of ampicillin (AMP), ciprofloxacin (CIP), ceftriaxone (CRO), sulfamethoxazole (SMZ), chloramphenicol (CL), and azithromycin (AZM) were detected and the genes and plasmids from azithromycin-resistant Salmonella were detected by Illumina hi-seq and Nanopore MinIone whole genome sequencing (WGS) using a map-based method, and the genomic background of these factors was evaluated using various bioinformatics tools. In total, 15 strains of nontyphoid Salmonella strains that were isolated (including S. typhimurium, S.London, S. Goldcoast, and S.Stanley) demonstrated resistance to azithromycin (minimum inhibitory concentration,MIC from 32 to >256 µg/mL), and the resistance rate was 3.08% (15/487). The sensitivity test to other antibiotics demonstrated 100% resistance to AMP, and the resistance to SMZ and CL was 86.7% and 80.0%, respectively. Through WGS analysis, all isolates were positive for a plasmid-encoded mphA gene. Plasmid incompatibility typing identified five IncFIB(K), five IncHI2/HI2A/Q1, two IncC, one IncHI2/HI2A/N, one IncR, one IncFII and one IncHI2/HI2A plasmids. Sequence analyses of plasmids revealed extensive homology to various plasmids or transposons in regions involved in plasmid replication/maintenance functions and/or in antibiotic resistance gene clusters. mphA is the main gene involved in azithromycin, a macrolide, and resistance to Salmonella. It is usually located on plasmids and easily spreads, hence posing a great threat to the current treatment of Salmonella infection. The plasmid sequence similarities suggest that the plasmids acquired resistance genes from a variety of enterica bacteria and underscore the importance of a further understanding of horizontal gene transfer among enterica bacteria.

Evaluation of antibiotic resistance in salmonella isolated from camel in a Najaf province

Non-Typhoidal Salmonellosis (NTS) is a form of salmonellosis that infect human from host-non adapted serotypes of salmonella, these salmonella strains are distributed in the environment, water and food via warm blooded animals causing gastroenteritis illnesses mostly, accompanied by high antibiotic resistance profile. In Najaf, camel meat considered one of the most used meat for human consumption. Yet, studies on distribution of salmonella and antibiotic resistance is scarce. In this research, (32) isolates of salmonella were isolated from camels in the province of Al-Najaf, using ordinary bacteriological methods, according to internationally known protocols for the cultivation and characterization of this bacteria. Then, all positive isolates subjected by antibiotic sensitivity test. The results showed a very high multidrug resistance profile was noticed among the isolated salmonella.

Salmonella serotypes isolated from Frozen chicken meat from Al- Najaf province markets

Salmonella is a zoonotic pathogen that can transfer from animals, contaminated animal products and contaminated water to humans. Non-Typhoidal Salmonella (NTS) are widely spread in wild, domestic animals and human being because it has no host restriction and this presents a broad spectrum of challenges to the public health causing gastroenteritis infections and dissemination of antibiotic resistance. Poultry is one of the most well-known host of NTS without apparent clinical manifestation, but study on overall salmonella population structure is limited. Therefore, a population structure study of salmonella serotypes in chicken meat is important to identify Salmonella serotypes and antibiotic resistance profile in Al-Najaf province. We targeted a novel genomic region that was previously approved for its ability to detect salmonella serotypes, strains of Salmonella in frozen chicken flesh, and antibiotic resistance patterns were conducted. 160 specimens from frozen chicken meat during the period extended from October 2020 to December 2020, 30 livers, 49 complete chickens, 25 thighs, and 56 breasts, collected from Al-Najaf province markets 44 (100%). The isolated samples were dominated by S. Infantis, 39/44 (88%) and 1/44 (2.2%) S.Enteritidis. Two non-salmonella bacteria, Citrobacter freundii 2/44 (4.5%) and Enterobacter cloacae 2/44 (4.5%) however.

Comparison of colourimetric loop-mediated isothermal amplification (LAMP), PCR and high-resolution melt curve analysis and culture-based diagnostic assays in the detection of three salmonella serotypes in poultry

ABSTRACT The accuracies of two molecular tests, PCR and loop-mediated isothermal amplification (LAMP) assay were compared with bacterial culture in detection of salmonella in poultry clinical samples. The icIR family transcriptional regulator gene was targeted and, out of 56 clinical specimens, 20 poultry field isolates were found positive for salmonella. Along with human isolates, reference strains of three different serovars, Salmonella Enteritidis (S. Enteritidis), S. Typhimurium and S. Infantis, were also tested. Eight different but genetically closely related bacterial genera (Klebsiella, Pseudomonas, Enterobacter, Campylobacter, Staphylococcus, Streptococcus, Escherichia and Pasteurella) were also used to evaluate the specificity of assay. The LAMP assay showed 80.8% sensitivity (95% CI, 0.66–0.95) and 100% specificity (95% CI, 0.71–1.00) when compared with microbiological culture and PCR, both with 100% sensitivity (95% CI, 0.87–1.00) and 100% specificity (95% CI, 0.71–1.00). High-resolution melt (HRM) curve analysis following PCR was able to differentiate between salmonella isolates based on their melting points, and all specimens were genotyped in three distinct HRM curve profiles. Each normalized melt curve profile represented one salmonella serotype and differences between the three melt profiles were correlated with nucleotide variations in the target gene sequences which demonstrated high discriminatory power of this technique. The colourimetric LAMP assay provided an alternative detection method capable of being used in the field, and showed analytical sensitivity for detection of 1 pg of salmonella DNA per reaction. The advantages and disadvantages of each test in detection of salmonella are discussed.

ISOLATION AND IDENTIFICATION OF SALMONELLA SPP FROM CHICKEN MEAT IN KURDISTAN REGION

Salmonella is a bacterial diseases that caused by many strains of salmonella. Salmonellosis is a disease occur in human and chicken, salmonellosis in poultry sometimes causes economic losses, also international trade can be affected if the salmonella presents in poultry products or poultry meat as in this case the infection can transmit to humans. Salmonella is a considered the most important and serious pathogen of bacteria that responsible of food borne infection all over the world. In Kurdistan region, Erbil city the common shapes of poultry are broilers and backyard. This study was done to inform the impact of salmonella epidemic chicken and the prevalence of salmonella's genetic types and serotypes among broilers and raw meat of chickens in Kurdistan, Iraq. Samples were collected from about fifty broiler chickens that suffered from diarrhea, fifty raw chickens meat and "30" patients that had diarrhea and signs of food poisoning. The serological identification of salmonella after isolation was "58.33%" salmonella enteritidis and "41.66%" salmonella typhimurium. Serotype salmonella enteritidis antigenic formula has "O" somatic antigen "1, 9, 12" and flagellar antigen "H", while the serotype salmonella typhimurium had the "O" somatic antigen "1, 4". The main infection source of humans includes; meat products that comes from consumption of contaminated meat of chicken. Scientifically, the control programs of impact of salmonella control are based on general procedures of hygiene on the campylobacter prevalence at the holding in broiler flocks and at the end of the process of slaughter on the broiler meat.

Monitoring of Salmonella infection of poultry for the period from 2016 to 2020

Salmonellosis is an acute intestinal infectious disease that belongs to the group of zoonoses that cause acute toxic infections in humans through the consumption of products of animal or plant origin contaminated with bacteria of the genus Salmonella spp. Salmonellosis is registered in all countries of the world, and our state is no exception.
 The aim of the research. Monitor salmonellosis pathogens in Ukrainian poultry farms and, on the basis of the obtained data, establish the relationship between avian and human diseases for salmonellosis.
 Materials and methods. The research was conducted in poultry farms in the North-Eastern region of Ukraine. Test systems from R-biopharm-Compact Dry SL and RIDA®STAMP SL were used for rapid diagnosis of bacterial microflora. To establish the salmonella serotype, the Spectate® salmonella test system was used, which is based on the use of latex strips coated with special antibodies to the corresponding serogroups of Salmonella A, B, C, D. Microbiological examination was performed washing from the walls, floor, eggshell, Brooder's cabinets and pathogens heart, liver, gallbladder of the dead bird. The relationship between isolated salmonella serovars from poultry was compared with salmonella isolated from humans according to the reporting form No. 40 “Zdorov”.
 Results. Various salmonella serovars were isolated from experimental farms of Ukraine, namely:
 Serovars such as S. Gallinarum–pullorum – 7.9 %, S. Enteritidis – 6.5 % were isolated from the meat and egg direction. From the meat direction, the following are: S. Tiphimurium – 12.5 %, S. Arizona – 6.0 %, S. Enteritidis – 3.5 %. From the breeding direction were also isolated from adult birds and hatching eggs in large quantities – S. Gallinarum–pullorum – 19.0 %, S. Tiphimurium – 17.8 %, S. Enteritidis – 10.1 %.
 The following species were separately isolated from the premises of the same farms in percentage terms: S. London – 1.7 %, S. Infantis – 1.5 % S. Bredeney – 1.4 %, S. Tsioque – 1.4 %, S Jawa – 1.2 %, S. Montevideo – 1.1 %, and 1 % each isolated S. Kentyki, S. Abony and S. Oxford. Pathogens of paratyphoid diseases were isolated from poultry and poultry products (S. Gallinarum–pullorum, S. Enteritidis, S. Tiphimurium) for the entire study period from 2016 to 2020 inclusive.
 Comparing the obtained data from state institutions of humane medicine, it should be noted that there was a tendency for the spread of salmonellosis among people associated with the consumption of poultry products.
 Conclusions. The obtained results indicate that salmonella infection is quite common among a number of poultry farms of different technological direction. In particular, bacteria of the genus Salmonella were isolated from meat, eggs and birds of different ages from the studied poultry houses in the North-Eastern region of Ukraine.
 As a result of monitoring of salmonellosis in Ukraine over the past five years, fluctuations in the incidence of salmonellosis among people ranged from a maximum of 74 % (2018) to a minimum of 30.1 % (2020), and for two years, there has been a reduction in cases due to the introduction of new international requirements and standards for product quality control at all stages of production of the Hazard Analysis and Critical Control Point (HACCP) system.

Evaluation of phytochemicals and activity index of some plant leaf extracts on typhoidal and non-typhoidal Salmonella isolates from selected hospitals in Bauchi, Nigeria

The pytochemicals and activity index evaluation was done for antibacterial activity of aqueous and methanol extracts of Cymbopogan citratus, Psidium guajava and Anacadium occidentale on clinical isolates of typhoidal and non typhoidal Salmonella. The typhoidal Salmonella isolates where S. typhi and S. paratyphi A, while non typhoidal was S. typhimurium. Well diffusion method was used. The activity of extracts was compared with that of orthodox drugs commonly used for the treatment of typhoid and non-typhoid salmonellosis thus; ciprofloxacin (10µg/Disc), amoxicillin (30µg/disc) and chloramphenicol (30µg/Disc) as standard. Phytochemical analysis of extracts was conducted using Trease and Evans methods of 2002. Both methanol and aqueous extracts had significant (p 1 on the serotypes indicating that Amoxacilin is resistant to salmonella serotypes tested. Using Ciprofloxacin as a standard, AI varies with serotypes S. paratypi A and S. typimurium have AI≤1 and S. typi have AI>1.Pyhtochemical screening of extract revealed the presence of some bioactive components like alkaloids saponin, tannins, anthraquinones steroid flavonoids glycosides. These properties determined the antimicrobial potential of the extracts. From the findings, it proved that plants crude extracts possess some potential as antibiotics and can be further studied to isolate the compound that is most active and formulated as drug against the disease.

Comparative Evaluation of the Effects of Binzalkonium Chloride, Iodine, Gluteraldehyde and Hydrogen Peroxide Disinfectants against Avian Salmonellae Focusing on Genotypic Resistance Pattern of the Salmonellae Serotypes toward Benzalkonium Chloride

ABSTRACT Using disinfectants in poultry houses is a common practice to ban the zoonotic pathogens like Salmonella. A major concern in using disinfectants is the emergence of bacteria strains that resist some disinfectants. This phenomenon is manifested in the resistance of some Salmonella serotypes against quaternary ammonium compounds. Such resistance is attributed to qacEΔ1 gene which may be possessed by some Salmonella serotypes. This work aimed to evaluate the resistance of Salmonella serotypes (S. Typhimurium, S. Infantis and S. Entiridis) against different disinfectants (benzalkonium chloride, iodine, gluteraldehyde and hydrogen peroxide). The effect of the disinfectants were evaluated by treatment of the bacteria with different concentrations (1:100, 200 and 400) at different temperatures and periods. Bacterial count was performed before and after the treatment. PCR for presence of qacEΔ1 gene was also performed before and after the treatment. The biocidal effect of the disinfectants found to be dependent on concentration, temperature and treatment period in addition to the type of the disinfectant. Hydrogen peroxide proved to be the most active agent followed by gluteradehyde, iodine and benzalkonium chloride. A link between the resistance against benzalkonium chloride and the existence of qacEΔ1 gene was proven in S. Typhimurium, whether treated or not treated with benzalkonium chloride.

СПЕКТР ЦИРКУЛИРУЮЩИХ СЕРОВАРИАНТОВ САЛЬМОНЕЛЛ В ПТИЦЕВОДЧЕСКИХ ХОЗЯЙСТВАХ

СПЕКТР ЦИРКУЛИРУЮЩИХ СЕРОВАРИАНТОВ САЛЬМОНЕЛЛ В ПТИЦЕВОДЧЕСКИХ ХОЗЯЙСТВАХ

Molecular Characterization of Different Salmonella Enterica Serotypes Isolated From Frozen Meat in Minoufiya Governorate

Salmonella is one of the most important causative agents of food poisoning and gastroenteritis in humans. This study spot highlights on isolation, identification and molecular characterization of salmonella serovars from imported frozen meat using the convential and modern molecular tools. Methods: A cross-sectional study was carried out on 100 samples of frozen meat collected from different supermarkets from Minufiya governorate, Egypt. Results: The prevalence of Salmonella were 6%. Serotyping of the obtained salmonella isolated revealed that Salmonella enteritidis, Salmonella typhimurium and Salmonella .paratyphi were the prevalent serotypes in the examined samples. S. typhimurium, only 3 samples (3%) ,Salmonella enteritidis was isolated from only 2 sample (2%) and S.paratyphi only 1 samples (1%). The application of conventional PCR for the six obtained isolates of Salmonella serotypes using universal gene (invA) was effective tool for identification and genotypic of pathogenic Salmonella serotypes. Conclusions: This study concluded that Salmonella is among the most important food borne pathogens worldwide contaminating a wide range of animal products including meat products. Also indicated that the cPCR was specific and rapid method for identification and genotyping of pathogenic salmonella serotypes.

Molecular Detection of Some Virulence Genes of Salmonella Serotypes Isolated From Poultry in Egypt

Salmonellosis is recognized as an important health problem and a major challenge in poultry industry worldwide. This study highlight the prevalence rate of salmonella serotypes in chickens as well as demonstration of some critical virulence genes by using specific primers sets in PCR reaction. Out of 100 freshly dead chickens, sixteen chicken samples were positive to Salmonella with 16% prevalence rate.The rate of recovery of Salmonellae from the different internal organs showed that the intestine is the most predication seat followed by, heart and liver with (8%), (6%) and (2%), respectively. Salmonella isolates (16) from chickens were serotyping using O and H antigen. The results revealed that S. Enteritidis was the most serotype found in 5 isolates (31.25%) followed by S.Infantis in 4 isolates (25%). Both S.Newlands and S.Kentucky found in 2 isolates 12.5%. Each of S.Ferruch, S.Weybridge and S.Naestved have one isolate (6.25%).The application of PCR for detection of invA, avrA and stn virulence genes in nine salmonella serovars revealed that these genes (invA, avrA and stn)were detected in 100% of tested strains except S. Naestved does not contain stn gene. Hence, this study recommends that PCR may be used for rapid and sensitive detection of Salmonella and help in understanding the role of these genes in pathogenicity of salmonella infection in poultry.

POTENTIAL MICROBIOLOGICAL HAZARDS IN BROILER CHICKENS DURING PROCESSING

Bacterial contamination of poultry carcasses during processing is undesirable and unavoidable consequently many potential microbiological hazards may be associated with preparation and processing of broiler carcasses. The aim of the present study was to obtain microbiological baseline data for broiler chickens at different processing steps in semi-automated small scale poultry abattoir located in Fayoum Governorate. Sampling was performed at 6 different processing points; after scalding, after defeathering, after evisceration, after washing, after chilling and after packaging. Carcasses were sampled by whole carcass rinse method and the rinsate was analyzed for coliforms (MPN), faecal coliforms (MPN), E. coli (MPN), Staphylococcus aureus count as well as isolation of Salmonellae and Campylobacter jejuni. Bacteriological examination recorded high bacterial counts (coliforms (MPN), faecal coliforms (MPN), E. coli (MPN) and Staphylococcus aureus count) in all sampling points. These counts increased during defeathering and evisceration then decreased after washing and chilling stage. E. coli, salmonella, Staphylococcus aureus and campylobacter jejuni were isolated by different percentages. The final packed carcasses were found to be heavily contaminated. The isolated E. coli serotypes were E. coli O1 , O127 , O44 , O78 ,O119 and O26 while the isolated salmonella serotypes were S. Virchow, S. Aba, S. Infantis and S. Kentucky.

Comparison of sporadic cases of Salmonella Typhimurium with other Salmonella serotypes in Castellon (Spain): case-case study

IntroductionSalmonella infections (SI) are common in Spain. The aim of this study was to appraise risk factors and the clinical characteristics of sporadic Salmonella Typhimurium infections compared with other sporadic salmonella serotype infections (OSI). MethodsFrom September 2014 to August 2015, a case-case study was carried out by the Epidemiology Division of the Public Health Centre of Castellon. Case 1 consisted of patients with sporadic S. Typhimurium infections, while case 2 comprised OSI patients, assessed according to the stool cultures analyzed by the Microbiology Laboratories of Hospital General de Castellon and Hospital de La Plana in Vila-real. Patients from detected outbreaks were not included. The salmonella serotype was identified by the National Centre of Microbiology (Madrid). ResultsThe total number of SI patients reported was 327, 242 of whom were studied (74.0%). 148 patients had sporadic S. Typhimurium infection and 64 had OSI, with median ages of 4 and 8.5 years, respectively. Sporadic S. Typhimurium infection patients presented more blood in feces and diarrhea episodes. Consumption of pork meat (OR=2.22; 95% CI 1.12–4.43), cold pork meats (OR=2.49; 95% CI 1.32–4.68) and playing in the dirt (OR=3.02; 95% CI 1.55–5.88), were associated with sporadic S. Typhimurium infection. In the 0–4 year-old group, the associated factors were consumption of cold pork meats, omelets and female gender. In the 5-year-old and over group, only playing in soil was associated with sporadic S. Typhimurium infection. ConclusionsThe consumption of pork and omelets, as well as playing in the dirt, were the main factors associated with infection. Children were most affected by sporadic S. Typhimurium infection.

Quasimetagenomics-Based and Real-Time-Sequencing-Aided Detection and Subtyping of Salmonella enterica from Food Samples.

Metagenomics analysis of food samples promises isolation-independent detection and subtyping of foodborne bacterial pathogens in a single workflow. The selective concentration of Salmonella genomic DNA by immunomagnetic separation (IMS) and multiple displacement amplification (MDA) shortened the time for culture enrichment of Salmonella-spiked raw chicken breast samples by over 12 h while permitting serotyping and high-fidelity single nucleotide polymorphism (SNP) typing of the pathogen using short shotgun sequencing reads. The herein-termed quasimetagenomics approach was evaluated on Salmonella-spiked lettuce and black peppercorn samples as well as retail chicken parts naturally contaminated with different serotypes of Salmonella Culture enrichment of between 8 and 24 h was required for detecting and subtyping naturally occurring Salmonella from unspiked chicken parts compared with 4- to 12-h culture enrichment when Salmonella-spiked food samples were analyzed, indicating the likely need for longer culture enrichment to revive low levels of stressed or injured Salmonella cells in food. A further acceleration of the workflow was achieved by real-time nanopore sequencing. After 1.5 h of analysis on a potable sequencer, sufficient data were generated from sequencing the IMS-MDA products of a cultured-enriched lettuce sample to enable serotyping and robust phylogenetic placement of the inoculated isolate.IMPORTANCE Both culture enrichment and next-generation sequencing remain time-consuming processes for food testing, whereas rapid methods for pathogen detection are widely available. Our study demonstrated a substantial acceleration of these processes by the use of immunomagnetic separation (IMS) with multiple displacement amplification (MDA) and real-time nanopore sequencing. In one example, the combined use of the two methods delivered a less than 24-h turnaround time from the collection of a Salmonella-contaminated lettuce sample to the phylogenetic identification of the pathogen. An improved efficiency such as this is important for further expanding the use of whole-genome and metagenomics sequencing in the microbial analysis of food. Our results suggest the potential of the quasimetagenomics approach in areas where rapid detection and subtyping of foodborne pathogens are important, such as for foodborne outbreak response and the precision tracking and monitoring of foodborne pathogens in production environments and supply chains.

Comparison of sporadic cases of Salmonella Typhimurium with other Salmonella serotypes in Castellon (Spain): case-case study

IntroductionSalmonella infections (SI) are common in Spain. The aim of this study was to appraise risk factors and the clinical characteristics of sporadic Salmonella Typhimurium infections compared with other sporadic salmonella serotype infections (OSI). MethodsFrom September 2014 to August 2015, a case-case study was carried out by the Epidemiology Division of the Public Health Centre of Castellon. Case 1 consisted of patients with sporadic S. Typhimurium infections, while case 2 comprised OSI patients, assessed according to the stool cultures analyzed by the Microbiology Laboratories of Hospital General de Castellon and Hospital de La Plana in Vila-real. Patients from detected outbreaks were not included. The salmonella serotype was identified by the National Centre of Microbiology (Madrid). ResultsThe total number of SI patients reported was 327, 242 of whom were studied (74.0%). 148 patients had sporadic S. Typhimurium infection and 64 had OSI, with median ages of 4 and 8.5 years, respectively. Sporadic S. Typhimurium infection patients presented more blood in feces and diarrhea episodes. Consumption of pork meat (OR=2.22; 95% CI 1.12–4.43), cold pork meats (OR=2.49; 95% CI 1.32–4.68) and playing in the dirt (OR=3.02; 95% CI 1.55–5.88), were associated with sporadic S. Typhimurium infection. In the 0–4 year-old group, the associated factors were consumption of cold pork meats, omelets and female gender. In the 5-year-old and over group, only playing in soil was associated with sporadic S. Typhimurium infection. ConclusionsThe consumption of pork and omelets, as well as playing in the dirt, were the main factors associated with infection. Children were most affected by sporadic S. Typhimurium infection.

Salmonella Thompson outbreak associated with consumption of chicken shawarma and the usefulness of genome sequencing in the investigation.

A sudden increase in Salmonella Thompson (S. Thompson) cases distributed throughout three border regions in the province of Quebec in November 2016 triggered a provincial investigation to identify a common source of contamination and to put the appropriate control measures into place. To report on the outbreak and to describe the use of genomic sequencing to identify the salmonella serotype responsible. A descriptive survey of all reported cases of Salmonella serogroup C1 that had occurred between October 1, 2016 and February 15, 2017 was conducted. A case definition was developed. Pulsed field gel electrophoresis supplemented by analyses of genome sequences using the single nucleotide variant phylogenomics method were used to demarcate and manage the outbreak. Eighteen cases of S. Thompson were identified through whole genome sequencing. The onset dates of symptoms for the 16 cases that presented enteric symptoms were November 21-December 2, 2016. Two cases that presented with atypical symptoms were not reported until February 2017. Among the 18 cases, 16 had eaten or probably eaten chicken shawarma at the same restaurant chain and nine of these cases ate it at the same restaurant. In total, five restaurants from this chain, spread throughout three border regions of Quebec, were identified. Outbreaks associated with chicken shawarma have been identified in the past. Efforts must be made to ensure that the owners of this type of restaurant know the contamination risk associated with this type of cooking and take the necessary steps to reduce this risk. The use of the genome sequencing method was very useful in defining the outbreak.

Resistance Testing of Salmonella Spp. Bacteria Isolated from Samples Derived from Poultry on Antimicrobial Drugs

In accordance with the Ordinance on establishing measures for the early detection, diagnosis, prevention of spreading, suppression and eradication of poultry infection by certain salmonella serotypes, antimicrobial agents are also applied in controlling salmonellosis of poultry, and the very treatment may lead to development of resistance to different drugs. The aim of this study was to determine the antimicrobial resistance of isolates, isolated from material originating from poultry farms during the one-year period (2014) in the territory of Nis and South Moravian epizootiological area. For the isolation and identification of causers, the standard microbiological methods according to EN ISO 6579: 2008 Annex D were used. Antimicrobial resistance is tested on 72 isolates of Salmonella spp. Examination of antimicrobial resistance was carried out by the disk diffusion method on Mueller-Hinton agar and interpreted according to the CLSI standard. The paper used thirteen types of antibiotic discs (Bioanalyse): enrofloxacin, norfloxacin, florfenicol, ceftiofur, amoxicillin, colistin, erythromycin, gentamicin, neomycin, kanamycin, flumequine, lincomicin /spectinomicin and sulfamethoxazole / trimethoprim. The results indicate the existence of a high percentage of resistance to some antimicrobial drugs. The greatest observed resistance was to erythromycin (68% at strain S. Enteritidis and 67% at S. Infantis), neomycin (61% at strain S. Enteritidis and 33% at S. Infantis), flumequine (32% at strain S. Enteritidis and 53% at S. infantis), strains of S. enteritidis showed a great resistance to kanamycin and sulfapreparations (23%) and colistin (20%), while the strains of S. infantis were resistant in a large percentage to amoxicillin (47%) and colistin (33%). There is slightly less resistance to other antibacterial drugs, and only a few isolates were resistant to quinolones and gentamycin. The emergence of resistant strains of Salmonella can result in the possibility of treating animals and humans and therefore a more rigorous implementation of measures for the suppression of salmonellosis in animals is required.

Isolation and Antimicrobial Resistance of Escherichia coli and Salmonella enterica subsp. enterica (O:6,8) in Broiler Chickens

Background: The third largest poultry flock in Northeast Brazil is located in Ceará State. Some pathogens are commonly disseminated in broiler chicken flocks, such as the bacteria from the Enterobacteriaceae family. Among these, some strains of Escherichia coli are frequently associated with different pathological manifestations in domestic animals, while bacteria from the genus Salmonella are considered the most frequent enteric pathogens reported causing foodborne infections in humans. Therefore, this study aimed to evaluate the prevalence and antimicrobial resistance of Salmonella sp. and Escherichia coli strains isolated from broiler chickens in the Metropolitan Region of Fortaleza city, Brazil.Materials, Methods & Results: Samples were collected from July-2014 to March-2015 in ten broiler chicken farms located in the Metropolitan Region of Fortaleza city, Brazil, with birds in pre-slaughter age. From each farm, 100 individual cloacal swabs were randomly collected from broilers independent of clinical status. Distinct methodologies were used in order to provide optimal isolation conditions for both the bacterial species. For Escherichia coli, the methodology consisted in enrichment with BHI broth, plating in EMB agar and biochemical identification, after which some isolates were maintained in nutrient agar for antimicrobial resistance evaluation. For the isolation of Salmonella sp., a standard method was used with pre-enrichment, selective enrichment, selective plating and biochemical identification steps. Antimicrobial susceptibility test (AST) was performed with disk diffusion technique and the following antibiotics were tested: ampicillin, ceftiofur, ciprofloxacin, trimethoprim-sulfamethoxazole, polymyxin B, gentamycin, cloranfenicol, tetracycline, azithromycin and fosfomycin. According to the methodology used, 95.9% of samples were positive for Escherichia coli and the most frequent resistance was to trimethoprim-sulfamethoxazole. Salmonella sp. was isolated from 0.2% of the samples, which were identified as the serotype Salmonella enterica subsp. enterica O:6,8. Both isolates presented the same antimicrobial resistance profile, which were resistant to six, out of ten tested antibiotics (trimethoprim-sulfamethoxazole, tetracycline, ciprofloxacin, azithromycin, chloramphenicol and ceftiofur).Discussion: The low prevalence of Salmonella observed in this study have also been reported by other studies performed in poultry farms in Ceará State, which suggests a good status for this pathogen in the local industry, however further efforts in order to eradicate this pathogen must be applied. The salmonella serotype detected in this study is rarely reported in the literature, especially from the poultry industry. In Brazil, the use of tetracyclines, quinolones and penicillins as feed additives or growth promoters is prohibited; however, a high resistance to drugs from these groups was detected. In addition, multidrug resistant E. coli isolates presented more elevated rates than other studies reported in the literature with antibiotics commonly used in the poultry industry and this may indicate an excessive use of these drugs in the production routine. These results should serve as a warning for surveillance programs to evaluate the incidence of these microorganisms as well as their antimicrobial resistance rates, which may be an important tool for control and prevention in meat poultry production.