[Molecular characterization of Listeria monocytogenes isolates from human and food sources in Argentina, 2018-2023]. / Caracterización molecular de aislamientos de Listeria monocytogenes de origen humano y alimentario en Argentina, 2018-2023.

Human listeriosis is an infectious disease caused by Listeria monocytogenes. The invasive form of this disease leads to a high rate of hospitalizations and fatality. The main mode of transmission is through contaminated ready-to-eat foods such as dairy, vegetables and meat products. The knowledge of the diversity and population dynamics of isolates collected from human and food sources is essential for the detection of clusters and the identification of common sites of infection. The aim of this study was the molecular characterization of L. monocytogenes isolates in Argentina. We sequenced a total of 63 isolates, 35 from human and 28 from food sources, collected between 2018 and 2023. Our genomic study divided the isolates into two lineages, four serogroups, 17 sequence types and 15 clonal complexes (CCs). The hypervirulent clone CC1 (lineage I; serogroup IVb) predominated in human and food samples. The phylogenomic analysis showed a high and possible epidemiological relationship between isolates from human and/or food sources, suggesting the presence of transmission chains in our country. These findings highlight the need to strengthen genomic surveillance of L. monocytogenes in Argentina. The identification of geographic distribution and characteristics of predominant and emerging clones from human and food sources might help to focus action plans and public health policies better directed at the control and prevention of listeriosis.

Resistencia antibiótica y aspectos clínicos de Neisseria gonorrhoeae anorrectal en un hospital de CABA, Argentina / Antimicrobial resistance and clinical aspects in anorectal Neisseria gonorrhoeae in a hospital in CABA, Argentina

La infección por Neisseria gonorrhoeae (NG) es considerada de alta prioridad en salud pública, por su capacidad para desarrollar resistencia a la mayoría de los antibióticos empleados para tratarla. La presentación anorrectal suele ser asintomática y frecuente en hombres que tienen sexo con hombres (HSH). En Argentina, se recomienda terapia antibiótica dual (ceftriaxona+azitromicina/doxiciclina) como primera línea empírica. Este estudio observacional y retrospectivo se realizó para evaluar el porcentaje de positividad de NG anorrectal, el perfil de sensibilidad a penicilina, tetraciclina, ciprofloxacina, ceftriaxona, cefixima y azitromicina, así como los aspectos clínicos-epidemiológicos de los pacientes atendidos entre 20/10/2015 y 20/03/2020 en consultorios coloproctológicos de un hospital público. Se detectaron 55/436 hisopados rectales positivos para NG (13%). El 95% era HSH y 71%, VIH+. En 18/55 NG fue la única infección. Las co-infecciones más frecuentes: HPV (38%) y C. trachomatis (35%). La sensibilidad a cefalosporinas de espectro extendido (CEE) y a azitromicina fueron 100% y 98%, respectivamente. Se observó la emergencia local de los primeros cinco aislamientos de NG anorrectal con sensibilidad reducida (SR) a CEE, el primer aislamiento con categoría no-sensible a azitromicina y otro con SR a azitromicina concomitantemente con SR a CEE. Aunque el uso de terapia empírica dual sigue siendo adecuado para nuestra institución, se observó la emergencia de aislamientos con SR y NS a las drogas de primera línea, evidenciando la importancia de la vigilancia epidemiológica a nivel local para definir los tratamientos empíricos.

Neisseria gonorrhoeae (NG) infection is considered a high public health priority because of its ability to develop resistance to most of the antibiotics used to treat it.The anorectal presentation is generally asymptomatic and frequent in men who have sex with men (MSM). In Argentina, dual therapy (ceftriaxone+azithromycin/doxycycline) is recommended as first line empiric therapy.This observational and retrospective study was conducted to evaluate the percentage of anorectal NG positivity, the susceptibility profile to penicillin, tetracycline, ciprofloxacin, ceftriaxone, cefixime and azithromycin, as well as the clinical-epidemiological aspects of patients attended between 20/10/2015 and 20/03/2020 in coloproctology of a public hospital.We detected 55/436 positive rectal swabs for NG (13%). 95% were MSM and 71% were PLHIV. In 18/55 NG was the only infection. The most frequent co-infections: HPV (38%) and C. trachomatis (35%).Susceptibility to extended-spectrum cephalosporins (ESCs) and azithromycin was 100% and 98%, respectively. Local emergence of the first five anorectal NG isolates with decreased susceptibility (DS) to ESCs, the first isolate with nonsusceptible category to azithromycin and another with DS to azithromycin concomitantly with DS to ESCs were observed.Although the use of dual empirical therapy continues to be adequate for our institution, the emergence of isolates with DS and NS to first-line drugs was observed, evidencing the importance of epidemiological surveillance at the local level to define empirical treatments

Sustained Transmission of Neisseria gonorrhoeae Strains with High-Level Azithromycin Resistance (MIC ≥ 256 µg/mL) in Argentina, 2018 to 2022.

Azithromycin combined with ceftriaxone is the recommended dual therapy for uncomplicated gonorrhea in many countries. Nevertheless, the increasing prevalence of azithromycin resistance compromises the effectiveness of this treatment strategy. From 2018 to 2022, we collected 13 gonococcal isolates with high-level azithromycin resistance (MIC ≥ 256 µg/mL) across Argentina. Whole-genome sequencing revealed that these isolates were mainly represented by the internationally spreading Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST) genogroup G12302, containing the 23S rRNA A2059G mutation (in all four alleles) together with mosaic mtrD and mtrR promoter 2 loci. This information is important to develop targeted public health policies to control the spread of azithromycin-resistant N. gonorrhoeae in Argentina and internationally. IMPORTANCE Azithromycin resistance in Neisseria gonorrhoeae has been increasing in numerous populations worldwide, which is of concern, as azithromycin is part of the recommended dual treatment in many countries. Here, we report 13 N. gonorrhoeae isolates with high-level azithromycin resistance (MIC ≥ 256 µg/mL). This study observed that high-level azithromycin-resistant gonococcal strains have shown sustained transmission in Argentina and are related to the successful international clone NG-MAST G12302. Genomic surveillance together with real-time tracing and data-sharing networks will be crucial in controlling the spread of azithromycin resistance in gonococcus.

Genomic Epidemiology of Azithromycin-Nonsusceptible Neisseria gonorrhoeae, Argentina, 2005-2019.

Azithromycin-nonsusceptible Neisseria gonorrhoeae strains are an emerging global public health threat. During 2015-2018, the prevalence of azithromycin-nonsusceptible gonococcal infection increased significantly in Argentina. To investigate the genomic epidemiology and resistance mechanisms of these strains, we sequenced 96 nonsusceptible isolates collected in Argentina during 2005-2019. Phylogenomic analysis revealed 2 main clades, which were characterized by a limited geographic distribution, circulating during January 2015-November 2019. These clades included the internationally spreading multilocus sequence types (STs) 1580 and 9363. The ST1580 isolates, which had MICs of 2-4 µg/mL, had mutations in the 23S rRNA. The ST9363 isolates, which had MICs of 2-4 or >256 µg/mL, had mutations in the 23S rRNA, a mosaic mtr locus, or both. Identifying the geographic dissemination and characteristics of these predominant clones will guide public health policies to control the spread of azithromycin-nonsusceptible N. gonorrhoeae in Argentina.

Testing of In Vitro Susceptibility of Neisseria gonorrhoeae to Azithromycin: Comparison of Disk Diffusion and Reference Agar Dilution Methods.

Azithromycin in combination with ceftriaxone is recommended as the first-line treatment for uncomplicated gonorrhea in many countries. Therefore, monitoring of azithromycin susceptibility of Neisseria gonorrhoeae isolates is essential. In 2019, the Clinical and Laboratory Standards Institute (CLSI) listed the MIC breakpoint for a susceptible-only category to azithromycin, but breakpoints for disk diffusion are not yet available. In this study, we evaluated the usefulness of disk diffusion for testing the susceptibility of N. gonorrhoeae isolates to azithromycin. A total of 189 clinical isolates susceptible and nonsusceptible to azithromycin were used. Agar dilution MICs were correlated with inhibition zone diameters of azithromycin disks (15-µg) manufactured by BBL and Oxoid. In addition, an interlaboratory study involving two clinical microbiology laboratories was conducted. There was a strong correlation between disk diffusion and agar dilution for BBL disks (r = -0.74; P < 0.001) and Oxoid disks (r = -0.75; P < 0.001). Using a zone diameter breakpoint of ≥27 mm (susceptible) and ≤26 mm (nonsusceptible) yielded good separation between susceptible and nonsusceptible isolates and the least number of discrepancies. Compared to agar dilution, disk diffusion showed high agreement and kappa values of 95.2% and 0.899 (P < 0.001) for BBL disks and 96.8% and 0.933 (P < 0.001) for Oxoid disks, respectively. Major and very major discrepancies were observed in isolates with azithromycin MICs (1 and 2 µg/ml, respectively) near to the breakpoint. These data illustrate that disk diffusion could be a reliable method in clinical laboratories to test susceptibility to azithromycin in N. gonorrhoeae isolates.

Use of Whole Genome Sequencing for the Molecular Comparison of Neisseria gonorrhoeae Isolates With Decreased Susceptibility to Extended Spectrum Cephalosporins From 2 Geographically Different Regions in America.

BACKGROUND: Neisseria gonorrhoeae isolates with reduced susceptibility or resistance to the recommended first-line antimicrobial therapy have been described in several countries. The purpose of this study was to use genome analyses to compare the molecular characteristics of N. gonorrhoeae isolates with decreased susceptibility to extended-spectrum cephalosporin from Ontario, Canada, and Argentina. METHODS: A total of 128 N. gonorrhoeae isolates, collected in 2015, were included. The susceptibility to penicillin G, tetracycline, ciprofloxacin, cefixime, ceftriaxone, and azithromycin was determined using the agar dilution method. Isolates were subjected to whole genome sequencing, and an in silico analysis was performed to identify antimicrobial resistance determinants and for genotyping. RESULTS: Decreased susceptibility to extended-spectrum cephalosporin was mainly associated with penA mosaic allele 34.001, together with an mtrR promoter A deletion and porB1b alterations G120K/A121N. N. gonorrhoeae multiantigen sequence typing ST1407 or closely related genotypes were identified circulating in both regions. CONCLUSIONS: An international multi-drug resistant clone of N. gonorrhoeae was associated with decreased susceptibility to extended-spectrum cephalosporin (ESC) in 2 different regions in America. Evidence of clonal dissemination of the organism in some regions suggests that the strength of surveillance programs and establishment of collaborative projects are essential.

Genome-based epidemiology and antimicrobial resistance determinants of Neisseria gonorrhoeae isolates with decreased susceptibility and resistance to extended-spectrum cephalosporins in Argentina in 2011-16.

OBJECTIVES: Our aim was to describe the molecular epidemiology and antimicrobial resistance determinants of isolates of Neisseria gonorrhoeae with decreased susceptibility and resistance to extended-spectrum cephalosporins (ESCs) in Argentina in 2011-16. METHODS: Gonococcal isolates (n=158) with decreased susceptibility and resistance to ESCs collected in 2011-16 across Argentina were subjected to WGS and antimicrobial susceptibility testing for six antimicrobials. RESULTS: In total, 50% of the isolates were resistant to cefixime, 1.9% were resistant to ceftriaxone, 37.3% were resistant to azithromycin and 63.9% of the isolates showed an MDR phenotype. Resistance and decreased susceptibility to ESCs was mainly associated with isolates possessing the mosaic penA-34.001, in combination with an mtrR promoter A deletion, and PorB1b amino acid substitutions G120K/A121N. Phylogenetic analysis revealed two main clades of circulating strains, which were associated with the N. gonorrhoeae multiantigen sequence typing (NG-MAST) ST1407 and closely related STs, and characterized by a high prevalence rate, wide geographical distribution and temporal persistence. CONCLUSIONS: N. gonorrhoeae isolates with decreased susceptibility and resistance to ESCs in Argentina have emerged and rapidly spread mainly due to two clonal expansions after importation of one or two strains, which are associated with the international MDR NG-MAST ST1407 clone. The identification of the geographical dissemination and characteristics of these predominant clones may help to focus action plans and public health policies to control the spread of ESC resistance in Argentina. Dual antimicrobial therapy (ceftriaxone plus azithromycin) for gonorrhoea needs to be considered in Argentina.

Comparison of disk diffusion and agar dilution methods for gentamicin susceptibility testing of Neisseria gonorrhoeae.

Gentamicin is a promising antibiotic for the treatment of multidrug-resistant gonorrhea. The aim of this study was to analyze the suitability and reliably of disk diffusion to monitor the susceptibility to gentamicin. We studied 237 Neisseria gonorrhoeae isolates obtained in 2013 and 2015. Reference MICs were correlated with inhibition zone diameters (in millimeters) of gentamicin 10 µg disks manufactured by BBL and Oxoid. The Pearson correlation between disk diffusion and agar dilution was r = -.68 (P < 0.001) for BBL disk and r = -.71 (P < 0.001) for Oxoid disk. No very major or major discrepancies were detected. However, a high percentage of minor discrepancies was observed (44.7%, BBL disk) and (21.9%, Oxoid disk). By adjusting the susceptible breakpoint to S ≥ 17 mm, the minor discrepancies rate was reduced to 19.4% (BBL disk) and 10.1% (Oxoid disk). The disk diffusion may be a screening method in clinical laboratories to detect the gentamicin susceptibility of N. gonorrhoeae.

Emergence and Spread of Neisseria gonorrhoeae Isolates With Decreased Susceptibility to Extended-Spectrum Cephalosporins in Argentina, 2009 to 2013.

BACKGROUND: The emergence of Neisseria gonorrhoeae strains with decreased susceptibility to cephalosporins represents a major concern globally. The aim of this study was to examine the phenotypic and molecular characteristics of N. gonorrhoeae isolates with decreased susceptibility to ceftriaxone and cefixime in Argentina. METHODS: A total of 1987 isolates were collected during 2009 and 2013. The susceptibility to penicillin G, tetracycline, ciprofloxacin, cefixime, ceftriaxone, and azithromycin was determined using the agar dilution method. The major extended-spectrum cephalosporin resistance determinants (penA, mtrR, and porB1b) were sequenced in 42 N. gonorrhoeae isolates that showed decreased susceptibility to ceftriaxone (minimum inhibitory concentration [MIC], 0.06-0.125 mg/L) and cefixime (MIC, 0.125-0.25 mg/L). Genotyping by N. gonorrhoeae multiantigen sequence typing (NG-MAST) was performed. RESULTS: Between 2009 and 2013, there was a shift in the modal MICs for ceftriaxone. Among the 42 isolates exhibiting decreased susceptibility to ceftriaxone and cefixime, 95.2% were resistant to penicillin G, 95.2% to tetracycline, 97.6% to ciprofloxacin, and 33.3% to azithromycin. Thirty-five (83.3%) of the 42 isolates had a mosaic penA allele XXXIV, which has been previously associated with resistance to ceftriaxone and cefixime as well as treatment failures. The isolates that contained the mosaic penicillin-binding protein 2 (PBP2) XXXIV were associated with NG-MAST ST1407 or closely related genotypes. CONCLUSIONS: In Argentina, N. gonorrhoeae isolates with decreased susceptibility to cefixime and ceftriaxone have now emerged, mostly due to the introduction of the internationally spread multidrug-resistant NG-MAST ST1407.

Prevalence of bla TEM-220 gene in Penicillinase-producing Neisseria gonorrhoeae strains carrying Toronto/Rio plasmid in Argentina, 2002 - 2011.

BACKGROUND: Penicillinase-producing Neisseria gonorroheae (PPNG) was first isolated in 1976. PPNG strains carrying bla TEM-1 and bla TEM-135 gene have been described in different countries. Recently, a novel bla TEM-220 allele was detected in PPNG isolates carrying Toronto/Rio plasmid. The prevalence and characteristics of TEM-220 strains worldwide are unknown, and therefore, it needs to be studied. The purpose of this study was to detect bla TEM-220 gene in PPNG strains possessing Toronto/Rio plasmid over a period of ten years in Argentina, and to evaluate the proportion of isolates producing non-TEM-220 containing the T539C substitution in the bla TEM allele. METHODS: One hundred and fifty one PPNG isolates carrying Toronto/Rio plasmid were studied between 2002 and 2011. A mismatch amplification mutation assay (MAMA) PCR was used to identify the T539C substitution in the bla TEM allele and a MAMA-PCR protocol was developed to detect the G547A substitution in the bla TEM-220. The reference agar dilution method of the Clinical and Laboratory Standard Institute (CLSI) was used for susceptibility testing to five ß-lactams antibiotics, ciprofloxacin, tetracycline and azithromycin. In all TEM-220-producing isolates, the whole bla TEM gene was sequenced and the isolates were typed using N. gonorroheae multiantigen sequence typing (NG-MAST). RESULTS: MAMA PCR successfully identified the G547A substitution in the bla TEM-220 allele. The proportion of isolates that possessed the bla TEM-220 allele was 2.6 %, and 93.2 % MAMA TEM-220 PCR-negative isolates showed the T539C substitution in the bla TEM gene. No differences in the susceptibility to five beta-lactam antibiotics tested were observed in PPNG isolates TEM-220-producing and PPNG isolates carrying the T539C substitution in the bla TEM gene. All TEM-220 isolates were indistinguishable by NG-MAST. CONCLUSION: This is the first study which shows the prevalence of bla TEM-220 in N. gonorrhoeae isolates carrying Toronto/Rio plasmid in Argentina. Although the bla TEM-220 allele does not appear to be associated with an extended spectrum beta-lactamase (ESBL) phenotype of resistance, a single nucleotide polymorphism added to the bla TEM-220 or bla TEM containing the T539C substitution could lead to the emergence of ESBL. Thus, it is imperative to investigate in surveillance programs, not only the plasmid type in PPNG isolates and the bla TEM allele associated, but phenotypical characteristics and geographical distribution of isolates.

Infección por neisseria gonorrhoeaey fenotipos de resistencia antimicrobiana,Mar del Plata, 2005-2010 / Neisseria gonorrhoeae infection and phenotypes of antimicrobial resistance, Mar del Plata, 2005-2010 / Infecção por Neisseria gonorrhoeae e fenótipos de resistência antimicrobiana, Mar del Plata, 2005-2010

Se realizó un estudio epidemiológico de tipo descriptivo retrospectivo, para determinar la tasa de infección por Neisseria gonorrhoeae y sus fenotipos de resistencia a los antimicrobianos, en 666 pacientes adultos de ambos sexos que asistieron al servicio al Servicio de ITS del Instituto Nacional de Epidemiología "Dr. Juan H. Jara"- ANLIS "Dr. Carlos G. Malbrán", de la ciudad de Mar del Plata, entre los años 2005-2010. Para el diagnóstico de infección por N. gonorrhoeae, las muestras fueron obtenidas por hisopados endocervicales e hisopados uretrales a varones y luego cultivadas. Los aislamientos fueron remitidos al Laboratorio Nacional de Referencia en ITS para completar el estudio de sensibilidad antimicrobiana. Se obtuvo una tasa de infección por N. gonorrhoeae del 12,2% [IC 95%: 9,83-14,95]. Los fenotipos de resistencia más prevalentes resultaron QRNG/CMRNG (resistencia a quinolonas conjuntamente con resistencia cromosómica a penicilina y tetraciclina), QRNG (resistencia a quinolonas), PPNG (cepa productora de penicilinasa) y CMTR (resistencia cromosómica a tetraciclina). En el marco de la vigilancia epidemiológica de las infecciones producidas por N. gonorrhoeae, el rol del laboratorio consiste no sólo en monitorear la incidencia de casos en la población sino también el perfil de resistencia a los antibióticos de uso terapéutico, a fin de controlar la enfermedad.

An epidemiological retrospective descriptive study was conducted in order to determine the rate of Neisseria gonorrhoeae infection and the antimicrobial resistance phenotypes in 666 adult patients of both sexes who attended the Sexually Transmitted Disease Service, at the National Institute of Epidemiology "Dr. Juan H. Jara"- ANLIS city of Mar del Plata, between the years 2005- 2010. For the diagnosis of N. gonorrhoeae infection, samples were obtained by endocervical swabs and urethral swabs in men. Microbiological growth on selective culture medium was identified using carbohydrate utilization. N. gonorrhoeae isolates were subsequently submitted to the National Reference Laboratory in STI for the study of antimicrobial susceptibility by determining the minimum inhibitory concentration. The N. gonorrhoeae infection rate was 12.2% [CI 95%: 9.83-14.95]. The most prevalent resistance phenotypes were QRNG/CMRNG (quinolone resistance in addition to chromosomal resistance to penicillin and tetracycline), QRNG (resistance to quinolone), PPNG (penicillinase producing strain) and CMTR (chromosomal resistance to tetracycline). As part of the epidemiological surveillance of infections by N. gonorrhoeae, the role of the laboratory is not only to monitor the incidence of cases in the population but also the antibiotic resistance profile for therapeutic use, in order to control the disease.

Um estudo epidemiológico de tipo descritivo retrospectivo foi realizado para determinar a taxa de infecção por Neisseria gonorrhoeae e seus fenótipos de resistência aos antimicrobianos em 666 pacientes adultos de ambos os sexos que compareceram no Serviço de ITS do Instituto Nacional de Epidemiología "Dr. Juan H. Jara, ANLIS "Dr. Carlos G. Malbrán" da cidade de Mar del Plata, entre os anos de 2005-2010. Para o diagnóstico de infecção por N. gonorrhoeae, as amostras foram obtidas por esfregaços endocervicais e esfregaços uretrais em homens e em seguida cultivadas. Os isolados foram encaminhados para o Laboratório Nacional de Referência em ITS para completar o estudo da sensibilidade antimicrobiana. Uma taxa de infecção de 12,2% em N. gonorrhoeae [IC 95%: 9,83-14,95] foi obtida. Os fenótipos de resistência mais prevalentes foram QRNG/CMRNG (resistência às quinolonas em conjunto com a resistência cromossômica à penicilina e tetraciclina), QRNG (resistência a quinolonas), PPNG (cepa produtora de penicilinase) e CMTR (resistência cromossômica à tetraciclina). Sob o controle epidemiológico das infecções produzidas por N. gonorrhoeae, o papel do laboratório é não só monitorar a incidência de casos na população, mas também o perfil de resistência aos antibióticos de uso terapêutico, visando a controlar a doença.