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We report here the complete genome of one Salmonella Agona strain isolated in 2017 from a dried milk powder in France.

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Multidrug-resistant (MDR) Salmonella enterica serovar Agona infections affect public health globally. This investigation aimed to ascertain the antimicrobial resistance profiles and molecular characteristics of Salmonella Agona isolates obtained from food-producing animals. A total of 209 Salmonella Agona isolates were recovered from mostly chickens (139 isolates), pigs (56 isolates), cattle (11 isolates), and ducks (3 isolates) between 2010 and 2020 in South Korea. In addition, these Salmonella Agona isolates were obtained from 25 slaughterhouses nationwide. Furthermore, this serotype suddenly increased in chickens in 2020. Salmonella Agona from chickens showed high resistance (69-83%) to ampicillin, streptomycin, tetracycline, trimethoprim/sulfamethoxazole, and chloramphenicol. Moreover, chicken/duck isolates (83.1%) showed significantly higher levels of MDR than cattle/pig isolates (1.5%). For molecular analysis by pulsed-field gel electrophoresis, infrared spectroscopy biotyping, and multilocus sequence typing in combination, a total of 23 types were observed. Especially two major types, P1-III-2-13 and P1-IV-2-13, comprised 59.3% of the total isolates spreading in most farms. Moreover, Salmonella Agona sequence type (ST)13 was predominant (96.7%) among three different STs (ST13, ST11, and ST292) widely detected in chickens (94.3%) in most farms located nationwide. Taken together, MDR Salmonella Agona in chickens might pose a potential risk to public health through direct contact or the food chain.

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Salmonella enterica Agona (S. Agona) and Salmonella enterica Saintpaul (S. Saintpaul) are among the emerging drug-resistant Salmonella in turkey production and processing. Rapid solutions to control emerging and uncommon serotypes such as S. Agona and S. Saintpaul are needed. This study tested pimenta essential oil (PEO) as a processing antibacterial against S. Agona and S. Saintpaul in experiments representative of different stages of turkey processing. The compound effectively reduced S. Agona and S. Saintpaul in nutrient broth studies and with mature biofilm assays. PEO was tested against a combination of S. Agona and S. Saintpaul in ground turkey meat and nonprocessed breast meat. In the first experiment with ground turkey, samples were inoculated with a mixture of S. Agona and S. Saintpaul (∼3 log10 CFU/g) and treated with PEO at different concentrations (0% PEO, 0.25% PEO, 0.5% PEO, 1% PEO, 2% PEO, and 2.5% PEO). In the second experiment with turkey breast, samples inoculated with ∼3 log10 CFU/g (SA+SP) were dipped in different concentrations of PEO with chitosan (CN) for 2 min. In both these experiments, samples were stored at 4°C, and Salmonella recovery was carried out at 0, 1, 3, 5, and 7 d. All experiments followed a completely randomized design and were repeated 6 times (n = 6). Statistical analysis was done using the PROC-ANOVA procedure of SAS. In the ground turkey meat, PEO at or above 2% reduced 2 log10 CFU/g of Salmonella by day 1. PEO at 2.5% in ground turkey meat resulted in enrichment-negative samples by 1 min, indicative of the rapid killing effect of the compound at a high concentration of PEO (P ≤ 0.05). A maximum reduction of 1.7 log10 CFU Salmonella/g of turkey breast meat was obtained after 2 min of dip treatment containing CN and 2.5% PEO. Results indicate that PEO could be used as a plant-based processing antibacterial against S. Agona and S. Saintpaul in turkey processing. Upscaling to plant-level studies is necessary before recommending its usage.

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Salmonella enterica subsp. enterica serovar Agona has a history of causing food-borne outbreaks and any emergence of multidrug-resistant (MDR) isolates in novel food products is of concern. Particularly, in food products frequently consumed without sufficient heating prior to consumption. Here, we report about the MDR isolate, 18-SA00377, which had been isolated from a dietary supplement in Germany in 2018 and submitted to the German National Reference Laboratory for Salmonella. WGS-based comparative genetic analyses were conducted to find a potential reservoir of the isolate itself or mobile genetic elements associated with MDR. As a phylogenetic analysis did not yield any closely related S. Agona isolates, either globally or from Germany, a detailed analysis of the largest plasmid (295,499 bp) was performed as it is the main carrier of resistances. A combined approach of long-read and short-read sequencing enabled the assembly of the isolate's chromosome and its four plasmids. Their characterization revealed the presence of 23 different antibiotic resistance genes (ARGs), conferring resistance to 12 different antibiotic drug classes, as well as genes conferring resistance to six different heavy metals. The largest plasmid, pSE18-SA00377-1, belongs to the IncHI2 plasmid family and carries 16 ARGs, that are organized as two distinct clusters, with each ARG associated with putative composite transposons. Through a two-pronged approach, highly similar plasmids to pSE18-SA00377-1 were identified in the NCBI database and a search for Salmonella isolates with a highly similar ARG resistance profile was conducted. Mapping and structural comparisons between pSE18-SA00377-1 and these plasmids and Salmonella isolates showed that both the plasmid backbone and identical or similar ARG clusters can be found not only in Salmonella isolates, originating mostly from a wide variety of livestock, but also in a diverse range of bacterial genera of varying geographical origins and isolation sources. Thus, it can be speculated that the host range of pSE18-SA00377-1 is not restricted to Salmonella and its spread already occurred in different bacterial populations. Overall, this hints at a complex history for pSE18-SA00377-1 and highlights the importance of surveilling multidrug-resistant S. enterica isolates, especially in novel food items that are not yet heavily regulated.

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Ovarian abscess is thought to be caused by ascending infection. We experienced a case with ovarian abscess caused by Salmonella. The patient had been monitored for endometrial cysts at an outpatient clinic. She visited our hospital emergently with chief complaints of persistent fever and diarrhea. Although sexually transmitted disease was not suspected, she was diagnosed with pelvic inflammatory disease due to her high C-reactive protein (CRP) level and started antibacterial treatment. Two days later, diagnostic laparoscopy was performed for persisting CRP elevation, and she was diagnosed with ovarian abscesses due to Salmonella infection. As the patient had regularly eaten soft-boiled eggs and chicken, an association with her eating habits was strongly suspected as a cause. When a patient has symptoms of enteritis, preceding tubo-ovarian abscess should be considered the possibility of Salmonella infection. Early diagnosis and treatment are necessary for fertility preservation, and minimally invasive diagnostic laparoscopy is useful.

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@#Introduction: Bacteria had long been known to have tumour-targeting and tumour inhibition capabilities and have re-emerged into the limelight of cancer research as a possible alternative treatment for solid tumours. Conventional therapies for solid tumours are either by surgery, chemotherapy, radiotherapy, which are very invasive and non-specific to the tumours and results in various adverse effects on the patients. Bacterial Mediated Tumour Therapy often utilises attenuated bacteria as therapeutic agents to ensure reduced pathogenicity of the strains. However, this often results in lower invasiveness towards the tumours itself. In this study, we studied the tumour inhibition capabilities of Salmonella Pathogenicity Island (SPI) attenuated Salmonella Typhimurium (S. Typhimurium) and Salmonella Agona (S. Agona), specifically with attenuation of sopB, sopD, and pipD genes. Methods: Balb/c mice bearing CT26 tumours were inoculated with S. Typhimurium and S. Agona, both unattenuated and ΔsopBΔsopDΔpipD attenuated strains. Tumour volumes were monitored daily. Organs and blood were collected for plasma liver enzyme analysis and histopathology studies on testis, liver, kidneys and brain. Results: The ΔsopBΔsopDΔpipD S. Agona treated group showed improved inhibition of tumour growth with 51.11% tumour volume reduction compared to unattenuated S. Agona. The ΔsopBΔsopDΔpipD strains have also shown lesser systemic effects as observed in plasma and histopathological studies) compared to its unattenuated counterparts. Conclusion: The present study showed that ΔsopBΔsopDΔpipD S. Agona has a great potential to be utilised as tumour therapeutic agent as it exerts lesser systemic effect while having similar tumour inhibition capabilities as the well-studied S. Typhimurium strain.

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Buffaloes and pigs play an important epidemiological roll in the Salmonella infection cycle, and asymptomatic animals can act as key component in the dissemination of the disease by horizontal, vertical, and cross-species transmission. Our study aimed and was able to confirm evidences of a cross-species transmission of Salmonella Agona between asymptomatic buffaloes and pigs. Also, we described Salmonella infection within the pig production phases, involving serotypes Agona, Senftenberg and Schwarzengrund. Rectal samples were collected from Jafarabadi buffaloes (n = 25) and Piau pigs (n = 32), located on a single farm. Salmonella Agona was isolated from lactating buffaloes, gilts, pregnant sows, and weaned pigs, Salmonella Schwarzengrund from lactating sows and Salmonella Senftenberg from gilts, pregnant sows, lactating sows, and weaned pigs. Pulsed-field Gel Electrophoresis protocol (PFGE) was performed and revealed four different profiles. Profile 1 (Salmonella Agona), isolated from a pregnant sow, a gilt and two lactating buffaloes, revealed a indistinguishable PFGE pattern, confirming evidences of potential cross-species transmission. Profile 2 (Salmonella Agona), 3 (Salmonella Senftenberg), and 4 (Salmonella Schwarzengrund), isolated from pigs, revealed important indistinguishable PFGE patterns, evidencing Salmonella infection within the pig production phases. Considering the epidemiological relevance of buffaloes and pigs in the cycle of Salmonella infection, confirmation of a potential cross-species transmission of Salmonella Agona and potential Salmonella infection within the pig production phases highlights the importance of the correct establishment of preventive health strategies in farms, in special the importance of avoiding contact between buffaloes and pigs, since cross-species transmission can occur, increasing the risk of spreading the disease.

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Our previous studies found that a dominant serovar of Salmonella enterica isolates from three farms raising broilers in 2014 and 2015 was serovar Agona and the number of Infantis isolates decreased (the serovar shift). In this study, 52 S. Agona strains which isolated between 1993 and 2008, were compared to the serovar shift clone by molecular epidemiology and phylogenetic analyses, using pulsed field gel electrophoresis and whole genome sequence analyses. Of the 52 strains, one strain isolated from a human case in 1995 was genetically identical to the serovar shift clone, even though it was isolated prior to the serovar shift. These results suggested that the S. Agona serovar shift clone had existed in a source other than chicken penetrated chicken population.

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The fight against cancer has been a never-ending battle. Limitations of conventional therapies include lack of selectivity, poor penetration and highly toxic to the host. Using genetically modified bacteria as a tumour therapy agent has gained the interest of scientist from the past few decades. Low virulence and highly tolerability of Salmonella spp. in animals and humans make it as the most studied pathogen with regards to anti-tumour therapy. The present study aims to construct a genetically modified S. Agona auxotroph as an anti-tumour agent. LeuB and ArgD metabolic genes in ΔSopBΔSopD double knockout S. Agona were successfully knocked out using a Targetron gene knockout system. The knockout was confirmed by colony PCR and the strains were characterized in vitro and in vivo. The knockout of metabolic genes causes significant growth defect in M9 minimal media. Quadruple knockout ΔSopBΔSopDΔLeuBΔArgD (BDLA) exhibited lowest virulence among all of the strains in all parameters including bacterial load, immunity profile and histopathology studies. In vivo anti-tumour study on colorectal tumour bearing-BALB/c mice revealed that all strains of S. Agona were able to suppress the growth of the large solid tumour as compared with negative control and ΔLeuBΔArgD (LA) and BDLA auxotroph showed better efficacy. Interestingly, higher level of tumour growth suppression was noticed in large tumour. However, multiple administration of bacteria dosage did not increase the tumour suppression efficacy. In this study, the virulence of BDLA knockout strain was slightly reduced and tumour growth suppression efficacy was successfully enhanced, which provide a valuable starting point for the development of S. Agona as anti-tumour agent.

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Salmonella enterica serovar Agona strains isolated from human cases were compared to strains that were derived from a clone caused a serovar shift in broilers. Pulsed field gel electrophoresis (PFGE) analysis with XbaI or BlnI digestion showed that three of seven strains from human case strains and most of the 81 strains from broilers were clustered in single complex in a minimum spanning tree (MST) reconstructed from the PFGE data. All the strains from human cases and 22 randomly selected strains from broilers were also analyzed by whole genome sequencing (WGS). Analysis of single nucleotide polymorphism (SNP) in the S. Agona core genes showed that four strains from human cases and all the strains from broilers were clustered in a maximum likelihood phylogenetic tree (ML tree) and an MST. These results indicated that the strains derived from the clone caused the serovar shift had already spread to humans. PFGE analysis with XbaI showed that four strains from broilers did not cluster with the other strains in an MST, though all those strains clustered in an ML tree and an MST reconstructed from SNP data. Moreover, three strains from broilers did not cluster in an MST reconstructed from PFGE with BlnI digestion, though those strains clustered in an ML tree and an MST reconstructed from SNP data. Therefore, it was suggested that S. Agona strains derived from a particular clone could not be traced by PFGE analysis but can be investigated by WGS analysis.

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On 1 December 2017, an outbreak of Salmonella Agona infections among infants was identified in France. To date, 37 cases (median age: 4 months) and two further international cases have been confirmed. Five different infant milk products manufactured at one facility were implicated. On 2 and 10 December, the company recalled the implicated products; on 22 December, all products processed at the facility since February 2017. Trace-forward investigations indicated product distribution to 66 countries.

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During May 2015, an increase in Salmonella Agona cases was reported from western Sydney, Australia. We examine the public health actions used to investigate and control this increase. A descriptive case-series investigation was conducted. Six outbreak cases were identified; all had consumed cooked tuna sushi rolls purchased within a western Sydney shopping complex. Onset of illness for outbreak cases occurred between 7 April and 24 May 2015. Salmonella was isolated from food samples collected from the implicated premise and a prohibition order issued. No further cases were identified following this action. Whole genome sequence (WGS) analysis was performed on isolates recovered during this investigation, with additional S. Agona isolates from sporadic-clinical cases and routine food sampling in New South Wales, January to July 2015. Clinical isolates of outbreak cases were indistinguishable from food isolates collected from the implicated sushi outlet. Five additional clinical isolates not originally considered to be linked to the outbreak were genomically similar to outbreak isolates, indicating the point-source contamination may have started before routine surveillance identified an increase. This investigation demonstrated the value of genomics-guided public health action, where near real-time WGS enhanced the resolution of the epidemiological investigation.

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The genome of a multidrug-resistant Salmonella Agona isolated from Larus audouinii (Audouin's gull) in Spain was examined. The isolate showed high levels of resistance to different antimicrobials, including third generation cephalosporins and fluoroquinolones, which is a public health concern as those being used to treat severe salmonellosis in humans. Whole genome sequencing revealed the strain being multilocus sequence type ST13, and eight resistance genes (aadA2, aadB, blaCTX-M-9,blaDHA-1, qnrA1, tetA, sul1 and dfrA16) belonging to seven antimicrobial classes were confirmed, as well as the presence of two plasmids. Migratory Audouin's gulls have the ability to cover long distances during annual movements. Therefore, they have the potential to disseminate multidrug-resistant Salmonella and resistance genes in the environment and over great geographic distances, contributing to the global dissemination of resistance genes.

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Se evaluaron retrospectivamente las características clínicas de 59 pacientes con aislamientos extraintestinales de Salmonella enterica no Typhi y la resistencia antibiótica entre 1988 y 2004. En el 95% de los casos se conocieron las condiciones de base del huésped que consistieron en: enfermedad oncohematológica (15), lupus eritematoso sistémico (11), síndrome de inmuno deficiencia adquirida (10), pacientes internados en neonatología (8), colecistitis (4) y otras (7). Las formas clínicas halladas fueron: bacteriemias primarias (26), bacteriemias secundarias a gastroenteritis (15), artritis séptica (7), infección urinaria (12), infección intraabdominal (5), meningitis (1) y pericarditis (1). En 29 pacientes se aisló Salmonella enterica no Typhi en más de una localización. El 15,8% y el 21,0% de 59 aislamientos presentaron resistencia a cefalosporinas de 3ª generación y a ampicilina, respectivamente. Salmonella Agona causó un brote epidémico en la sala de cuidados intensivos de neonatología, aislándose de sangre en 5 pacientes y en uno también de LCR. Este serotipo fue multirresistente. Imipenem, trimetroprima-sulfometoxazol y ciprofloxacina fueron los antibióticos más activos (CIM 50: 0,5; 0,5 y 0,002 µg/mL, respectivamente).

The clinical and antimicrobial resistance of 59 patients with extraintestinal infection by nontyphi Salmonella was retrospectively evaluated from 1988 to 2004. In 95% of the patients there were underlying diseases, which included: oncohematologic disease (15), systemic lupus erythematosus (11), acquired immunodeficiency disease (AIDS) (10), patients of neonatal units (8), colecistitis (4), and other disorders (7). The clinical manifestation were primary bacteremias (26), bacteremias secondary to gastroenteritis (15), arthritis (7), urinary tract infections (12), abdominal infections (5), meningitis (1) and pericarditis (1). In 29 patients Salmonella enterica no Typhi was isolated in more than one site. Resistance to third generation cephalosporins was observed in 15,8 % of these isolates, and resistance to ampicillin in 21,0%. Salmonella Agona was isolated from an outbreak in neonatal ICU; the organism was recovered from blood samples of five neonates and in one patient also isolated from CSF. These isolates showed multiresistance. Imipenem, trimetoprim-sulfametoxazol and ciprofloxacin were the three most active antimicrobial agents (MIC 50: 0.5, 0.5 and 0.002 µg/mL, respectively).

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