Resumen La bacteria Escherichia coli causa la colibacilosis en animales de granja que actúan como reservorios de cepas patógenas. La resistencia antimicrobiana de E. coli productor de betalactamasas de espectro extendido [BLEE] es un grave problema de salud pública y se puede atribuir a factores relacionados con el consumo de alimentos y el contacto con animales domésticos. El objetivo de este estudio fue determinar la presencia y patrones de resistencia antimicrobiana de E. coli BLEE aislado en muestras fecales provenientes de bovinos productores de leche de la provincia de Pichincha. Se analizaron un total 182 muestras de heces de bovinos: 112 muestras de bovinos faenados en el Camal Metropolitano de la provincia de Pichincha y 70 muestras de la colección de la Unidad de Investigación de Enfermedades Transmitidas por Alimentos y Resistencias a los Antimicrobianos [UNIETAR], se realizó el aislamiento de E. coli BLEE, la identificación bioquímica y pruebas de resistencia a los principales antibióticos utilizados. Se logró identificar 93 muestras positivas a E. coli BLEE (51 %), el análisis fenotípico reveló que los antibióticos amoxicilina más ácido clavulánico, cefepime, ceftazidima, ciprofloxacina, amikacina y tetraciclina presentaron porcentajes de resistencia mayores al 80 %. Además, se observó una baja resistencia a la nitrofurantoína, cefoxitin y ertapenem, mientras que ningún aislado fue resistente a la tigeciclina. El 100 % de los aislados presentaron fenotipos de multirresistencia y el patrón más frecuente estuvo compuesto por 7 familias de antibióticos. En conclusión, estos resultados sugieren que E. coli originaria de bovinos lecheros podría ser un reservorio de genes BLEE.

Abstract Escherichia coli causes colibacillosis in farm animals that act as reservoirs for pathogenic strains. Antimicrobial resistance of E. coli producing Extended Spectrum Beta-lactamases [ESBL] is a serious public health problem that can be attributed to factors related to food consumption and contact with domestic animals. This study aimed to determine the presence and patterns of antimicrobial resistance of ESBL E. coli isolated from fecal samples from dairy cattle in the Pichincha province. A total of 182 bovine feces samples were analyzed, 112 samples from cattle slaughtered at the official slaughterhouse in Quito-Pichincha province, and 70 samples from the collection of the Foodborne Diseases and Antimicrobial Resistance Research Unit [UNIETAR]. The isolation of ESBL E. coli, biochemical identification, and resistance tests using the main antibiotics were carried out at UNIETAR. It was possible to identify 93 positive samples for ESBL E. coli (51%), phenotypic analysis revealed that antibiotics such as amoxicillin plus clavulanic acid, cefepime, ceftazidime, ciprofloxacin, amikacin, tetracycline, presented resistance higher than 80%. Furthermore, low resistance to nitrofurantoin, cefoxitin, and ertapenem was observed, while no isolate was resistant to tigecycline. One hundred percent of the isolates presented multi-resistance phenotypes, with the most frequent pattern being composed of 7 families of antibiotics. In conclusion, these results suggest that E. coli from dairy cattle origin could be an important reservoir of ESBL genes.

Abstract Introduction. In Mexico, 85 % of the crops are white maize, 10 % white yellow and 5 % other colors. Biochemical studies of pigmented native maize have been carried out, but there is little information on colour-improved maize. Objective. To evaluate the phytochemical content and antimicrobial capacity of two pigmented maize hybrids grown at the Tecnológico Nacional de México-Valle del Yaqui. Materials and methods. Extraction of biocompounds from a portion of the maize grains grown in southern Sonora, Mexico, during the summer-winter 2020 cycle was performed on red and purple maize hybrids. A thirty-part solution of ethanol, acetic acid and water was used. Proximal composition, anthocyanin, polyphenol and flavonoid content were determined by standardized methods. Antioxidant capacity was determined by ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methodologies. In addition, antimicrobial capacity and minimum inhibitory concentration were determined. Results. Purple maize showed higher content of anthocyanins (340.98±5.21 mg/100 g) and polyphenols (173.68±24.23 mg gallic acid/100 g) compared to red maize. Flavonoids are more abundant in purple maize (575.10±27.88 mg quercetin/100 g). Both maize hybrids exhibited more than 50 % antioxidant capacity against ABTS and DPPH radicals. Regarding antimicrobial activity, higher inhibition was observed for Escherichia coli and Salmonella (18 % and 47 %), and lower for Staphylococcus aureus and Shigella (19 % and 34 %) compared to gentamicin. Conclusion. Purple maize showed higher content of anthocyanins, polyphenols and flavonoids. Both hybrids had an antioxidant capacity higher than 50 %. Antimicrobial activity was higher against E. coli and Salmonella, than against S. aureus and Shigella.

Resumen Introducción. En México, el 85 % de las siembras son maíces blancos, el 10 % amarillo blancos y el 5 % de otros colores. Se han realizado estudios bioquímicos de maíces nativos pigmentados, pero existe poca información en maíces mejorados en color. Objetivo. Evaluar el contenido de fitoquímicos y capacidad antimicrobiana de dos híbridos de maíz pigmentado cultivados en el Tecnológico Nacional de México, Valle del Yaqui. Materiales y métodos. En granos de híbridos de maíz de color rojo y morado se realizaron las extracciones de biocompuestos a partir de una porción del maíz cultivado en el sur de Sonora, México, durante el ciclo verano-invierno 2020. Se utilizó una solución de etanol, ácido acético y agua en una proporción de treinta partes. La composición proximal, contenido de antocianinas, polifenoles y flavonoides se determinaron mediante métodos estandarizados. La capacidad antioxidante se evaluó mediante las metodologías ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) y DPPH (2,2-difenil-1-picrilhidracilo). Además, se determinó la capacidad antimicrobiana y la concentración mínima inhibitoria. Resultados. El maíz morado presentó mayor contenido de antocianinas (340,98±5,21 mg/100 g) y polifenoles (173,68±24,23 mg de ácido gálico/100 g), en comparación con el maíz rojo. Los flavonoides son más abundantes en el maíz morado (575,10±27,88 mg de quercetina/100 g). Ambos híbridos de maíz exhibieron una capacidad antioxidante superior al 50 % frente a los radicales ABTS y DPPH. Respecto a la actividad antimicrobiana, se observó mayor inhibición para Escherichia coli y Salmonella (18 % y 47 %), y menor para Staphylococcus aureus y Shigella (19 % y 34 %) en comparación con la gentamicina. Conclusión. El maíz morado mostró mayor contenido de antocianinas, polifenoles y flavonoides. Ambos híbridos tuvieron una capacidad antioxidante mayor al 50 %. La actividad antimicrobiana fue mayor contra E. coli y Salmonella, que contra S. aureus y Shigella.

Abstract Surveillance indicators of the quality of water for human consumption in the Amazon were analysed from 2016 to 2020 using 185,528 samples from 11 microregions. Of the samples analysed, 93.20% were from urban areas, 66.65% were from the public water supply system (WSS), 31.02% were from the Collective Alternative Solution-CAS, and 2.33% from the Individual Alternative Solution-IAS. There was an increase in the number of records by the WSS, with a downwards trend and fluctuations in records for the CAS and the IAS. The quality indicators of chemical and physical parameters for urban areas were higher than those for rural areas and traditional communities. Most of the samples presented pH values below the recommended level. In the quantification of microbiological parameters, a higher presence of total coliforms and E. coli was identified in samples from rural areas and in traditional communities. In conclusion, there were inadequacies in the chemical, physical and microbiological parameters as well as problems related to the supply, storage and surveillance of water distributed for human consumption. These findings indicate the need to build an agenda for public management to address water insecurity and its likely effects on food insecurity in the region. 201 202 185528 185 528 185,52 1 microregions 9320 93 20 93.20 6665 66 65 66.65 WSS , (WSS) 3102 31 02 31.02 SolutionCAS, SolutionCAS Solution CAS, Solution-CAS 233 2 33 2.33 SolutionIAS. SolutionIAS IAS Solution-IAS communities level E conclusion region 18552 18 52 185,5 932 9 93.2 666 6 66.6 (WSS 310 3 0 31.0 23 2.3 1855 5 185, 93. 66. 31. 2.

Resumo Analisaram-se indicadores de vigilância da qualidade da água para consumo humano no Amazonas, de 2016 a 2020, utilizando 185.528 amostras provenientes de 11 microrregiões. Das amostras analisadas, 93,20% são da área urbana, 66,65% provinham do sistema público (SAA), 31,02% da Solução Alternativa Coletiva (SAC) e 2,33% da Solução de Alternativa Individual (SAI). Observou-se aumento do número de registros pelo SAA, com tendência de queda e oscilações de registros para a SAC e a SAI. Os indicadores de qualidade dos parâmetros químicos e físicos da área urbana foram superiores aos das áreas rurais e de comunidades tradicionais. A maior parte das amostras apresentou valores de pH abaixo do recomendado. Na quantificação dos parâmetros microbiológicos, identificou-se maior presença de coliformes totais e E.coli na área rural e em comunidades tradicionais. Em conclusão, verificaram-se inadequações nos parâmetros químicos, físicos e microbiológicos, assim como problemas relativos ao abastecimento, armazenamento e à vigilância da água distribuída para consumo humano. Tais achados indicam a necessidade de construir uma agenda, pela gestão pública, para o enfrentamento da insegurança hídrica e seus prováveis efeitos sobre a insegurança alimentar existente na região. Analisaramse Analisaram se Amazonas 201 2020 185528 185 528 185.52 1 microrregiões analisadas 9320 93 20 93,20 6665 66 65 66,65 SAA , (SAA) 3102 31 02 31,02 (SAC 233 2 33 2,33 SAI . (SAI) Observouse Observou tradicionais recomendado microbiológicos identificouse identificou Ecoli E coli conclusão verificaramse verificaram abastecimento agenda pública região 202 18552 18 52 185.5 932 9 93,2 666 6 66,6 (SAA 310 3 0 31,0 23 2,3 (SAI 1855 5 185. 93, 66, 31, 2,

ABSTRACT: The Amazonian manatee (Trichechusinunguis) is an aquatic mammal threatened with extinction. However, few studies have investigated the pathogens in this species, which may affect both animal and human health. This study aimed to evaluate the frequency, distribution, and patterns of antimicrobial susceptibility of Staphylococcus spp. and Escherichiacoli colonizing the nasal and rectal cavities of Amazonian manatees kept in captivity at the National Institute for Amazonian Research (INPA) in the state of Amazonas, Brazil. Rectal and nasal swabs from 44 manatees of different ages were used in this study. The genus Staphylococcus was isolated from the nasal swabs of 32 (72.7%) animals, with two individuals harboring more than one species of Staphylococcus. S. sciuri was the most commonly isolated species. Resistance to penicillin was observed in 13 (40.6%) isolates, more frequent than the other antimicrobials tested (P = 0.01). E. coli was isolated from the rectal swabs of all animals, with phylogroup B1 being the most frequent among the strains obtained (P = 0.0008). Four isolates (6.8%) were positive for virulence factors, three of which were classified as enterotoxigenicE. coli (ETEC) and one as enteropathogenicE. coli (EPEC). To our knowledge, this is the first study to evaluate Staphylococcus spp. and E. coli in Amazonian manatee samples. This study revealed nasal colonization by Staphylococcus spp., mainly S. sciuri, and diarrheagenicE. coli isolates, including antimicrobial-resistant strains. ABSTRACT Trichechusinunguis (Trichechusinunguis extinction However health frequency distribution spp INPA (INPA Amazonas Brazil 4 3 72.7% 727 72 7 (72.7% animals S 1 40.6% 406 40 6 (40.6% P 0.01. 001 0.01 . 0 01 0.01) E B 0.0008. 00008 0.0008 0008 0.0008) 6.8% 68 8 (6.8% factors enterotoxigenicE ETEC (ETEC enteropathogenicE EPEC. EPEC (EPEC) knowledge samples diarrheagenicE antimicrobialresistant resistant 72.7 (72.7 40.6 (40.6 00 0.0 0000 0.000 000 6.8 (6.8 (EPEC 72. (72. 40. (40. 0. 0.00 6. (6. (72 (40 (6 (7 (4 (

RESUMO: O peixe-boi amazônico (Trichechus inunguis) é um mamífero aquático ameaçado de extinção. Apesar disso, poucos estudos investigaram patógenos nessa espécie que podem impactar a saúde animal e humana. O objetivo deste estudo foi avaliar a frequência, distribuição e padrões de suscetibilidade antimicrobiana de Staphylococcus sp. e Escherichia coli pertencentes à microbiota nasal e retal de peixes-bois da Amazônia mantidos em cativeiro no Instituto Nacional de Pesquisas da Amazônia (INPA) (Amazonas, Brasil). Foram utilizados suabes retais e nasais de 44 peixes-boi de diferentes idades. O gênero Staphylococcus foi isolado do swab nasal de 32 (72,7%) animais, sendo que dois indivíduos apresentaram mais de uma espécie de Staphylococcus. S. sciuri foi a espécie mais comumente isolada. A resistência à penicilina foi observada em 13 (40,6%) isolados, sendo mais frequente do que os outros antimicrobianos testados (P = 0,01). Escherichia coli foi isolada dos suabes retais de todos os animais, sendo o filogrupo B1 o mais frequente entre as cepas obtidas (P = 0,0008). Quatro isolados (6,8%) foram positivos para fatores de virulência, três dos quais foram classificados como E. coli enterotoxigênica (ETEC) e um isolado como E. coli enteropatogênica (EPEC). Este é o primeiro estudo a avaliar Staphylococcus spp. e E. coli de amostras de peixe-boi da Amazônia. O estudo revelou colonização nasal por Staphylococcus sp., principalmente S. sciuri, e por isolados diarreiogênicos de E. coli, incluindo cepas resistentes a antimicrobianos. RESUMO peixeboi peixe boi Trichechus inunguis extinção disso humana frequência sp peixesbois peixes bois INPA (INPA Amazonas, Amazonas (Amazonas Brasil. Brasil . Brasil) 4 peixesboi idades 3 72,7% 727 72 7 (72,7% animais S 1 40,6% 406 40 6 (40,6% P 0,01. 001 0,01 0 01 0,01) B 0,0008. 00008 0,0008 0008 0,0008) 6,8% 68 8 (6,8% virulência E ETEC (ETEC EPEC. EPEC (EPEC) spp 72,7 (72,7 40,6 (40,6 00 0,0 0000 0,000 000 6,8 (6,8 (EPEC 72, (72, 40, (40, 0, 0,00 6, (6, (72 (40 (6 (7 (4 (

Abstract: Bacteremia is a major cause of morbidity and mortality in patients with cancer and episodes of high-risk febrile neutropenia (HRFN). Objective: To identify the frequency of microorganisms isolated from blood cultures (BC) and their antimicrobial resistance (R) profile in children with HRFN, compared with the same data from previous studies of the same group. Method: Prospective, multicenter, epidemiological surveillance study of microorganisms isolated from BC in patients under 18 years of age, from 7 PINDA network hospitals, between 2016 and 2021. Results: 284 episodes of HRFN with positive BC were analyzed out of 1091 enrolled episodes (26%). Median age 7.2 years [3.0-12.3]. The main isolates were gram-negative bacilli (GNB) 49.2%, gram-positive cocci (GPC) 43.8%, and fungi 3.6%. The most frequently isolated microorganisms were viridans group Streptococci (VGS) (25.8%), Escherichia coli (19.8%), Pseudomonas spp. (11.2%), Klebsiella spp. (10.9%), and coagulase negative Staphylococci (CoNS) (10.9%). There was an increase in R to third-generation cephalosporins (p = 0.011) in GNB and to oxacillin in CoNS (p = 0.00), as well as a decrease in R to amikacin in non-fermenting GNB (p = 0.02) and to penicillin in VGS (p = 0.04). Conclusion: VGS is the main agent isolated in BC from pediatric patients with cancer and episodes of HRFN, followed by E. coli, Pseudomonas spp., and Klebsiella spp. Having epidemiological surveillance of microorganisms isolated from BC and their antimicrobial R profile is essential to favor the rational use of antimicrobials.

Resumen: Las bacteriemias son una causa importante de morbimortalidad en pacientes con cáncer y episodios de neutropenia febril de alto riesgo (NFAR). Objetivo: Determinar la frecuencia de microorganismos aislados de hemocultivos (HC) y su perfil de resistencia (R) antimicrobiana en niños con NFAR, comparado con iguales datos de estudios previos del mismo grupo. Material y Método: Estudio prospectivo, multicéntrico, de vigilancia epidemiológica de microorganismos aislados de HC en pacientes menores de 18 años, en siete hospitales de la red PINDA, 2016-2021. Resultados: Se analizaron 284 episodios de NFAR con HC positivos de 1091 episodios enrolados (26%). Mediana de edad 7,2 años [3,0-12,3]. Los aislados principales fueron bacilos gramnegativos (BGN) 49,2%, cocáceas grampositivas (CGP) 43,8% y hongos 3,6%, con mayor frecuencia de Streptococcus grupo viridans (SGV) (25,8%), Escherichia coli (19,8%), Pseudomonas spp. (11,2%), Klebsiella spp. (10,9%) y Staphylococcus coagulasa negativa (SCN) (10,9%). Hubo aumento de R a cefalosporinas de tercera generación en enterobacterales (p = 0,011) y de R a oxacilina en SCN (p = 0,00), así como disminución de R a amikacina en BGN no fermentadores (p = 0,02) y de R a penicilina en SGV (p = 0,04). Conclusión: SGV es el principal agente aislado en HC de pacientes pediátricos con cáncer y episodios de NF, seguido de E. coli, Pseudomonas spp. y Klebsiella spp. Contar con vigilancia epidemiológica de microorganismos aislados de HC y su perfil de R antimicrobiana es esencial para favorecer el uso racional de antimicrobianos.

Abstract This study analyzed the impact of leachate from a temporary landfill on soil microbial diversity in Tingo María, Huánuco region, Peru. Three treatments were used: untreated soil (S), addition of stream water (T0), leachate (T1), and leachate treated by coagulation and flocculation (T2), with 828.5 ml/week added in three weekly doses. Soil samples were collected from the Reserved Forest of the Universidad Nacional Agraria de la Selva. Twenty-one randomly distributed soil samples were taken and homogenized for analysis. Soil quality parameters measured included sand, clay, silt, texture, pH, organic matter, nitrogen, phosphorus and potassium. As for microorganisms, viable aerobes, lactobacilli, actinomycetes, fungi, nitrogen-fixing bacteria, and Escherichia coli were quantified using specific culture and counting methods for each of them. To evaluate the impact of the leachate on microbial diversity, equity indices (Shannon and inverse Simpson), dominance indices (complementary Simpson and Berger Parker) and the percentage composition of each microorganism per treatment were used. An ANOVA was performed to estimate differences in microbial diversity, with a Tukey test at a significance level of α = 0.05. The study showed that leachates affect soil microbial diversity, reducing equity and increasing the dominance of certain species such as E. coli. They also alter physicochemical parameters, decreasing organic matter and nitrogen but increasing other elements such as phosphorus and potassium. This could have implications for soil health and functionality.

Resumen Las infecciones por Entamoeba se producen en todo el mundo, con mayor frecuencia en países de bajo nivel socioeconómico y salud pública deficiente. Dado que se ha reconocido a Entamoeba histolytica como la única especie patógena del género, es importante realizar un diagnóstico diferencial respecto de otras especies de Entamoeba morfológicamente idénticas. Este estudio tuvo como objetivos determinar la prevalencia de especies de Entamoeba en 2 poblaciones de Argentina, realizar su diagnóstico diferencial por PCR y caracterizar los aislados de Entamoeba secuenciando un fragmento del gen SSU ARNr. Se obtuvieron 493 muestras fecales seriadas de individuos de las provincias de Buenos Aires (n=210) y Misiones (n=283). Las muestras se examinaron por métodos convencionales (sedimentación de formalina-etil acetato y flotación de Willis) y mediante PCR específicas para diferenciar especies de Entamoeba. Los aislamientos de Entamoeba se caracterizaron por secuenciar un fragmento del gen SSU ARNr. La prevalencia general de la infección por Entamoeba fue del 12,4% y fue mayor en Buenos Aires que en Misiones (14,8 vs. 10,6%). Se informó por primera vez un caso de Entamoeba histolytica en Buenos Aires, confirmado por PCR y análisis de secuencia. Además, se registraron nuevos datos genéticos sobre Entamoeba coli y Entamoeba dispar. El análisis filogenético reveló una congruencia entre las características morfológicas y las secuencias del gen SSU ARNr. A través de este estudio, hemos sumado información acerca de la distribución de estas especies en nuestro país y en la región de las Américas.

Abstract Entamoeba infections occur worldwide, with higher frequency in countries of low socioeconomic status and poor public health. Since Entamoeba histolytica has long been recognized as the only pathogenic species, making a differential diagnosis of other morphologically identical Entamoeba is important. This study aimed to determine the prevalence of Entamoeba species in two populations from Argentina, make a differential diagnosis by PCR and characterize Entamoeba isolates at the SSU rRNA gene. A total of 493 serial fecal samples were obtained from individuals in the provinces of Buenos Aires (n=210) and Misiones (n=283). Samples were examined by conventional methods (formalin-ethyl acetate and Willis flotation) and specific PCRs to differentiate Entamoeba species. Entamoeba isolates were characterized by sequencing a fragment of the SSU rRNA gene. The overall prevalence of Entamoeba infection was 12.4%, being more prevalent in Buenos Aires than in Misiones (14.8% vs. 10.6%). A case of E. histolytica confirmed by PCR and sequence analysis was reported for the first time in Buenos Aires. Moreover, new genetic data on Entamoeba coli and Entamoeba dispar were recorded. The phylogenetic analysis revealed a congruence between morphological characteristics and SSU rRNA gene sequences. This study increases the amount of information on the distribution of these species in Argentina and the region of the Americas.

Biologically active compounds in most spices possess antimicrobial and other important biomedical properties. There have been huge demands for natural immunity boosters (spices and herbs), considering the recent global pandemic and challenges relating to drug-resistant pathogens. This study was designed to compare the efficacy of ginger, garlic and turmeric spices against some pathogenic microorganisms. Aqueous extraction of spices, antimicrobial sensitivity and minimum inhibitory concentration tests were done using standard microbiological methods. Bioactive compounds were estimated using the gas chromatography-mass spectrometry (GC-MS) method. Aqueous extracts of ginger inhibited the growth of all test isolates except Streptococcus pneumoniae, with inhibition zones ranging between 0.9 mm and 13.5 mm. Escherichia coli, S. pneumoniae and Haemophilus influenzae were resistant to turmeric extracts, while the extract of garlic inhibited only four of the test pathogens. Inhibition zones for turmeric ranged between 4.4 mm and 10.9 mm, while those for garlic were between 4.7 mm and 11.5 mm. All the spice extracts did not inhibit microbial growth at 10-40%. An antibiotic spectrum indicated that Bacillus sp. was resistant to all but one, nitrofurantoin, which also inhibited the growth of almost all pathogens, except H. influenzae, with zones ranging between 10.5 mm and 11.6 mm. All test pathogens were resistant to cloxacillin except E. coli (10.6 mm). The major phyto-active compounds present in ginger are 2-Butanone,4-(4-hydroxy-3-methoxyphenyl), 1,3-Cyclohexadiene and 1-(4-Hydroxy-3-methoxyphenyl). SIGNIFICANCE: Conclusively, ginger, turmeric and garlic have varied inhibitory activities against diverse organisms, indicating their antimicrobial properties; however, ginger showed a higher inhibitory effect and more diverse antimicrobial property amongst selected isolates. Furthermore, certain bioactive compounds of biomedical importance were present. We therefore recommend the use of these spices as alternative natural food preservatives against spoilage organisms, as well as potential natural sources for bioactive compounds in drug development against pathogens.

FtsZ is a bacterial divisome protein responsible for Z-ring formation in cytokinesis. Characterization of Escherichia coli FtsZ protein oligomers (EcFtsZ) in native conditions is defiance because the protein is found as a multi-oligomer in self-association-dissociation equilibrium. We characterize the trimeric state of EcFtsZ through native PAGE, gel filtration chromatography and sucrose gradient techniques combined with chemical cross-linking. The filtration results indicate that the EcFtsZ trimer has a molecular mass of 131 kDa and a filtration friction coefficient (Rs/Rmin) equal to 1.9, while the theoretical filtration friction coefficient (fn/f1) calculated for a linear trimer yielded a value equal to 1.8 very close to the experimental value. On the other side, formaldehyde-crosslinked EcFtsZ showed a band 128 kDa recognized by anti-FtsZ antibodies, and a sedimentation friction coefficient (Smax/S20,w) equal to 1.9, while the theoretical sedimentation friction coefficient (Sn/S1) calculated for a triangular trimer was equal to the experimental value. These results suggest that EcFtsZ has two homotrimeric structures (linear and triangular). Finally, we report the aggregation of EcFtsZ at micromolar concentrations of Capsaicin without GTP and Mg2+.

La FtsZ es una proteína del divisoma bacteriano responsable de la formación del anillo Z en la citocinesis. La caracterización de los oligómeros de la proteína FtsZ de Escherichia coli (EcFtsZ) en su estado nativo es un desafío porque la proteína se encuentra como un multioligómero en equilibrio de autoasociación-disociación. Nosotros caracterizamos el estado trimérico de la EcFtsZ a través de técnicas de PAGE nativo, cromatografía de filtración en gel y gradiente de sacarosa combinada con entrecruzamiento químico. Los resultados por filtración indican que el trímero de EcFtsZ tiene una masa molecular de 131 kDa y un coeficiente de fricción por filtración (Rs/Rmin) igual a 1.9, mientras que el coeficiente de fricción por filtración teórico (fn/f1) calculado para un trímero lineal arrojó un valor igual a 1.8 que está muy cercano al valor experimental. Por otro lado, la EcFtsZ entrecruzada con formaldehído presentó una banda de 128 kDa reconocida por anticuerpos anti-FtsZ y un coeficiente de fricción por sedimentación (Smax/S20,w) igual a 1.9, mientras que el coeficiente de fricción por sedimentación teórico (Sn/S1) calculado para un trímero triangular resultó igual a valor experimental. Estos resultados sugieren que la EcFtsZ tiene dos estructuras homotriméricas (lineal y triangular). Finalmente, reportamos la agregación de la EcFtsZ a concentraciones micromolares de Capsaicina sin GTP y Mg2+.

Abstract: Bacteremia is a major cause of morbidity and mortality in patients with cancer and episodes of high-risk febrile neutropenia (HRFN). Objective: To identify the frequency of microorganisms isolated from blood cultures (BC) and their antimicrobial resistance (R) profile in children with HRFN, compared with the same data from previous studies of the same group. Method: Prospective, multicenter, epidemiological surveillance study of microorganisms isolated from BC in patients under 18 years of age, from 7 PINDA network hospitals, between 2016 and 2021. Results: 284 episodes of HRFN with positive BC were analyzed out of 1091 enrolled episodes (26%). Median age 7.2 years [3.0-12.3]. The main isolates were gram-negative bacilli (GNB) 49.2%, gram-positive cocci (GPC) 43.8%, and fungi 3.6%. The most frequently isolated microorganisms were viridans group Streptococci (VGS) (25.8%), Escherichia coli (19.8%), Pseudomonas spp. (11.2%), Klebsiella spp. (10.9%), and coagulase negative Staphylococci (CoNS) (10.9%). There was an increase in R to third-generation cephalosporins (p = 0.011) in GNB and to oxacillin in CoNS (p = 0.00), as well as a decrease in R to amikacin in non-fermenting GNB (p = 0.02) and to penicillin in VGS (p = 0.04). Conclusion: VGS is the main agent isolated in BC from pediatric patients with cancer and episodes of HRFN, followed by E. coli, Pseudomonas spp., and Klebsiella spp. Having epidemiological surveillance of microorganisms isolated from BC and their antimicrobial R profile is essential to favor the rational use of antimicrobials.

Resumen: Las bacteriemias son una causa importante de morbimortalidad en pacientes con cáncer y episodios de neutropenia febril de alto riesgo (NFAR). Objetivo: Determinar la frecuencia de microorganismos aislados de hemocultivos (HC) y su perfil de resistencia (R) antimicrobiana en niños con NFAR, comparado con iguales datos de estudios previos del mismo grupo. Material y Método: Estudio prospectivo, multicéntrico, de vigilancia epidemiológica de microorganismos aislados de HC en pacientes menores de 18 años, en siete hospitales de la red PINDA, 2016-2021. Resultados: Se analizaron 284 episodios de NFAR con HC positivos de 1091 episodios enrolados (26%). Mediana de edad 7,2 años [3,0-12,3]. Los aislados principales fueron bacilos gramnegativos (BGN) 49,2%, cocáceas grampositivas (CGP) 43,8% y hongos 3,6%, con mayor frecuencia de Streptococcus grupo viridans (SGV) (25,8%), Escherichia coli (19,8%), Pseudomonas spp. (11,2%), Klebsiella spp. (10,9%) y Staphylococcus coagulasa negativa (SCN) (10,9%). Hubo aumento de R a cefalosporinas de tercera generación en enterobacterales (p = 0,011) y de R a oxacilina en SCN (p = 0,00), así como disminución de R a amikacina en BGN no fermentadores (p = 0,02) y de R a penicilina en SGV (p = 0,04). Conclusión: SGV es el principal agente aislado en HC de pacientes pediátricos con cáncer y episodios de NF, seguido de E. coli, Pseudomonas spp. y Klebsiella spp. Contar con vigilancia epidemiológica de microorganismos aislados de HC y su perfil de R antimicrobiana es esencial para favorecer el uso racional de antimicrobianos.

Resumo As bactérias foram isoladas de amostras de suco de maçã fresco de lojas de três diferentes localidades de Lahore. A análise de amostras dos mercados Liberty, Anarkali e Yateem khana mostram diferentes níveis de contaminação. Havia bactérias patogênicas e não patogênicas em todas as amostras e foram identificadas pelos testes morfológicos e bioquímicos. A maioria dos plasmídeos de bactérias patogênicas tinha 4 kb em seu tamanho molecular. A ribotipagem do sequenciamento do gene do RNA ribossômico 16S foi realizada para confirmar a cepa de Helicobacter pylori e Gluconobacter oxydans. A maior sensibilidade de 210 mm foi mostrada por Enterobacter sp. contra disco de azteromisina (15µg) enquanto Micrococcus sp. foi altamente resistente a todos os antibióticos aplicados. A resistência a antibióticos de bactérias patogênicas também foi verificada contra extratos de plantas de Ricinus communis, todos os patógenos bacterianos isolados foram resistentes, mas apenas E. coli foi inibida em 300µl dos extratos. A presença de bactérias patogênicas nas amostras de suco de maçã deveu-se à contaminação da água de esgoto na água potável, enquanto algumas dessas bactérias patogênicas vieram da árvore da maçã e outras de armazéns de frutas.

Abstract Bacteria were isolated from samples of Fresh Apple juices from shops of three different localities of Lahore. Analysis of samples from Liberty, Anarkali and Yateem khana Markets show different levels of contamination. There were pathogenic and non-pathogenic bacteria in all samples and were identified by the morphological and biochemical tests. Most of the plasmids of pathogenic bacteria were 4kb in their molecular size. Ribotyping of 16S ribosomal RNA gene sequencing was done to confirm Helicobacter pylori strain and Gluconobacter oxydans. The highest sensitivity of 210mm was shown by Enterobacter sp. against Aztheromysine disk (15µg) while Micrococcus sp. was highly resistant against all of the Antibiotics applied. The antibiotic resistance of pathogenic bacteria was also checked against Ricinus communis plant's extracts, all isolated bacterial pathogens were resistant but only, E.coli was inhibited at 300µl of the extracts. Presence of pathogenic bacteria in Apple juice samples was due to contamination of sewage water in drinking water while some of these pathogenic bacteria came from Apple's tree and other from store houses of fruits.

Resumo O presente estudo envolve a análise química e bacteriológica de água de diferentes fontes, ou seja, furo, poços, garrafa e torneira, dos distritos de Peshawar, Mardan, Swat e Kohat da província de Khyber Pakhtunkhwa (KP), Paquistão. De cada distrito, 50 amostras de água (10 amostras de cada fonte), independentemente do status urbano e rural, foram coletadas dessas fontes e analisadas para sulfatos, nitratos, nitritos, cloretos, sólidos solúveis totais e coliformes (E. coli). Os resultados indicaram que a maioria das fontes de água tinha uma contagem inaceitável de E. coli, ou seja, > 34 UFC / 100 mL. As amostras positivas para E. coli foram elevadas no distrito de Mardan, seguido por Kohat, Swat e distrito de Peshawar. Além disso, algumas fontes de água também foram contaminadas quimicamente por diferentes fertilizantes inorgânicos (nitratos/nitritos de sódio, potássio), mas em níveis seguros, enquanto os resíduos agrícolas e industriais (compostos de cloreto e sulfato) estavam em níveis inseguros. Entre todos os distritos, a qualidade da água foi considerada comparativamente mais deteriorada nos distritos de Kohat e Mardan do que nos distritos de Peshawar e Swat. Essas fontes de água química e bacteriologicamente impróprias para beber podem causar problemas à saúde humana.

Abstract The present study involves the chemical and bacteriological analysis of water from different sources i.e., bore, wells, bottle, and tap, from Peshawar, Mardan, Swat and Kohat districts of Khyber Pakhtunkhwa (KP) province, Pakistan. From each district, 50 water samples (10 samples from each source), regardless of urban and rural status, were collected from these sources and analysed for sulphates, nitrates, nitrites, chlorides, total soluble solids and coliforms (E. coli). Results indicated that majority of the water sources had unacceptable E. coli count i.e.> 34 CFU/100mL. E. coli positive samples were high in Mardan District, followed by Kohat, Swat and Peshawar district. Besides this, the some water sources were also chemically contaminated by different inorganic fertilizers (nitrates/nitrites of sodium, potassium) but under safe levels whereas agricultural and industrial wastes (chloride and sulphate compounds) were in unsafe range. Among all districts, the water quality was found comparatively more deteriorated in Kohat and Mardan districts than Peshawar and Swat districts. Such chemically and bacteriologically unfit water sources for drinking and can cause human health problems.

ABSTRACT: The domestic and international market is increasingly seeking for foods with reduced chemical additives. The viability using a versatile product as mechanically separate meat (CMS) to produce safety food is necessary to expand the product market. Hansenulawingei strain was selected, a yeast known as a killer toxin for poultry CMs application. The first part of this study consisted of defining the best growth condition for yeast, static or agitation, as well as the time either growth. The result obtained was the medium without agitation and 108 hours of growth at room temperature. The extract was subjected to the atomization drying process and the pulverized extract obtained was evaluated in vitro for minimum inhibitory concentration (MIC) tests for Salmonellasp, Staphylococcus positive coagulase, E. coli and mesophilic bacteria. Proximal composition, which the most relevant values to be analyzed were directed to total proteins that were 4.26g% (± 0.66) and 4.37g% (± 0.01) for agitation and unrestrained system, respectively. In the result of MIC, dry extracts were efficient in controlling the growth of all tested bacteria, including Salmonella spp. In the in situ tests, in which mechanically separated meat was raw materials to determine the antimicrobial factor action against food pathogens the concentration of 0.083g/ml of dry extract in its raw form obtained an inhibition factor as satisfactory as healing salts traditionally applied by the carneous industry. The viability of its growth and concentration for drying, can be considered a viable antimicrobial with good prospects for bio-conservative action. ABSTRACT additives CMS (CMS selected application 10 temperature MIC (MIC Salmonellasp coagulase E bacteria composition 426g g 4 26g 4.26g ± ( 0.66 066 0 66 437g 37g 4.37g 0.01 001 01 system respectively spp 0083gml gml 083g ml industry bioconservative bio conservative 1 0.6 06 6 0.0 00 0.

RESUMO: O mercado nacional e internacional busca cada vez mais por alimentos com reduzido teor de aditivos químicos. Estudar a viabilidade de um novo método de conservação natural de um produto tão versátil como a carne mecanicamente separada (CMS) se faz necessário para expandir o mercado do produto, seguindo as tendências das demandas alimentares dos consumidores atuais. Portanto foi selecionada uma cepa de Hansenula wingei, uma levedura conhecida pela produção de toxina killer para aplicação em CMS de aves. A primeira parte deste estudo consistiu em definir qual a melhor condição de crescimento para a levedura, consistindo este em ser estático ou em agitação, bem como o tempo no qual haveria o maior pico de crescimento da mesma. O resultado obtido foi o meio sem agitação e com 108 horas de crescimento a temperatura ambiente. Na sequência o extrato foi submetido ao processo de secagem por atomização e o pó obtido foi avaliado in vitro para testes de concentração mínima inibitória (MIC) para Salmonella sp, Staphylococcus coagulase positivo, E. coli e Mesofilos aeróbicos e definição de sua composição proximal, a qual os valores mais relevantes a serem analisados eram direcionados para proteínas totais que foram 4.26g% (±0.66) e 4.37g% (±0.01) para sistema com agitação e sem agitação, respectivamente. Como resultado de MIC, os extratos secos mostraram-se eficientes no controle do crescimento de todas as bactérias testadas, inclusive Salmonella spp. Nos testes in situ, no qual a carne mecanicamente separada foi a matéria prima testada para determinar a ação fator antimicrobiano contra patógenos de alimentos a concentração de 0.083g/ml do extrato seco, em sua forma bruta, obteve um fator de inibição tão satisfatório quanto aos sais de cura aplicados tradicionalmente pela indústria cárnea e pela viabilidade de seu crescimento e concentração por secagem, pode ser considerado um antimicrobiano viável e com boas perspectivas para ação bioconservante. RESUMO químicos (CMS atuais wingei aves mesma 10 ambiente MIC (MIC sp positivo E proximal 426g g 4 26g 4.26g ±0.66 066 0 66 (±0.66 437g 37g 4.37g ±0.01 001 01 (±0.01 respectivamente mostraramse mostraram testadas spp situ 0083gml gml 083g ml seco bruta bioconservante 1 ±0.6 06 6 (±0.6 ±0.0 00 (±0.0 ±0. (±0. ±0 (±0 ± (± (

Abstract Emergence of plasmid mediated colistin and extended spectrum β-lactamases (ESBL) resistant genes has been impacted the efficacy of colistin and β-lactams drugs like 3rd, 4th generation cephalosporin. Current study was aimed to investigate antimicrobial resistance genes (ARGs) among Escherichia coli isolates from meat producing commercial broilers in Pakistan. Two hundred (n=200) fecal samples were collected during January-2018 to August-2019. For isolation of E. coli, pink colonies on MacConkey agar were transferred to EMB agar. Metallic sheen color colonies were tested biochemically using API-20E kit. The molecular identification of E. coli (n=153) was targeted by amplification of uid gene through polymerase chain reaction (PCR) and different ARGs i.e. gentamicin, streptomycin, tetracycline, colistin, β-lactams drugs, quinolone and ampicillin followed by sequence analysis. Genotypically, followed by phenotypically of resistant ARGs of isolated PCR-confirmed E. coli (153) shoed resistant against gentamicin (aac(3)-IV), streptomycin (aadA1), tetracycline (tetA), colistine (mcr-1), ampicillin (bla-TEM) and bla-CTX-M were 86%, 88%, 86%, 88%, 83% & 77% respectively. 33/38 (86%) of the isolate was positive for quinolone resistance. Colistine (mcr-1), ESBLs (bla-TEM) and (bla-CTX-M) resistance genes were 88%, 83% and 77% respectively. About 33 isolated E. coli harbored the both mcr-1 and ESBLs genes. All of E. coli isolates were found sensitive to ceftriaxone (CTX-30) and imipenem (IMP-10). The Isolated E. coli showed single or multi clade decadency. The E. coli and ARGs sequences showed single or multi clade decadency. This is first comprehensive study from Pakistan that described the molecular evidences of ARGs and their co-existence in single isolates originated from commercial poultry. Commercial chicken (Broilers) can act as melting pot of antibiotic resistance genes for human being. It is alarming situation for surveillance of antibiotic resistance program because of more regulated prescription of antimicrobial agents in Pakistan βlactamases β lactamases ESBL (ESBL βlactams lactams 3rd rd th cephalosporin (ARGs n=200 n200 n 200 (n=200 January2018 January 2018 January-201 August2019. August2019 August 2019. 2019 August-2019 E API20E APIE API 20E kit n=153 n153 153 (n=153 PCR (PCR ie i e i.e analysis Genotypically PCRconfirmed confirmed (153 aac3IV, aac3IV aacIV aac 3 IV , (aac(3)-IV) aadA1, aadA1 aadA (aadA1) tetA, tetA (tetA) mcr1, mcr1 mcr 1 (mcr-1) blaTEM bla TEM (bla-TEM blaCTXM CTX M 86 86% 88 88% 83 77 respectively 3338 38 33/3 (86% (bla-CTX-M mcr- CTX30 30 (CTX-30 IMP10. IMP10 IMP 10 . (IMP-10) decadency coexistence co existence poultry Broilers (Broilers being n=20 n20 20 (n=20 January201 201 January-20 August201 August-201 n=15 n15 15 (n=15 (15 (aac(3)-IV (aadA1 (tetA (mcr-1 8 7 333 33/ (86 CTX3 (CTX-3 IMP1 (IMP-10 n=2 n2 2 (n=2 January20 January-2 August20 August-20 n=1 n1 (n=1 (1 (aadA (mcr- (8 (CTX- (IMP-1 n= (n= January2 January- August2 August-2 ( (mcr (CTX (IMP- (n August- (IMP

Resumo O surgimento de colistina mediada por plasmídeo e genes de resistência a β-lactamases de espectro estendido (ESBL) afetou a eficácia de medicamentos colistina e β-lactâmicos, como as cefalosporinas de 3ª e 4ª geração. O presente estudo teve como objetivo investigar genes de resistência antimicrobiana (ARGs) entre isolados de Escherichia coli em frangos de corte comerciais no Paquistão. Duzentas (n = 200) amostras fecais foram coletadas durante janeiro de 2018 a agosto de 2019. Para o isolamento de E. coli, colônias rosas em ágar MacConkey foram transferidas para ágar EMB. As colônias de cores de brilho metálico foram testadas bioquimicamente usando o kit API-20E. A identificação molecular de E. coli (n = 153) foi direcionada pela amplificação do gene uid através da reação em cadeia da polimerase (PCR) e diferentes ARGs, ou seja, gentamicina, estreptomicina, tetraciclina, colistina, medicamentos β-lactâmicos, quinolona e ampicilina, seguido de análise de sequência. Genotipicamente, seguido por fenotipicamente de ARGs resistentes de E. coli isoladas foram confirmadas por PCR (153) como resistente contra gentamicina (aac(3)-IV), estreptomicina (aadA1), tetraciclina (tetA), colistina (mcr-1), ampicilina (bla-TEM) e bla-CTX-M, demonstrando resultados de 86%, 88%, 86%, 88%, 83% e 77%, respectivamente. Cerca de 33/38 (86%) do isolado foi positivo para resistência às quinolonas. Os genes de resistência à colistina (mcr-1), ESBLs (bla-TEM) e (bla-CTX-M) foram 88%, 83% e 77%, respectivamente. Cerca de 33 E. coli isoladas continham os genes mcr-1 e ESBLs. Todos os isolados de E. coli foram considerados sensíveis à ceftriaxona (CTX-30) e imipenem (IMP-10). A E. coli isolada apresentou decadência de um ou vários clados. As sequências de E. coli e ARGs apresentaram decadência de um ou vários clados. Este é o primeiro estudo abrangente do Paquistão que descreveu as evidências moleculares de ARGs e sua coexistência em isolados únicos originados de aves comerciais. Dessa forma, é possível concluir que o Frango comercial (Broilers) pode atuar como caldeirão de genes de resistência a antibióticos para o ser humano. É uma situação alarmante para a vigilância do programa de resistência a antibióticos devido à prescrição mais regulamentada de agentes antimicrobianos no Paquistão βlactamases β lactamases ESBL (ESBL βlactâmicos, βlactâmicos lactâmicos, lactâmicos β-lactâmicos ª geração (ARGs n 200 201 2019 E EMB API20E. API20E APIE API 20E. 20E API-20E 153 (PCR seja sequência Genotipicamente (153 aac3IV, aac3IV aacIV aac 3 IV , (aac(3)-IV) aadA1, aadA1 aadA (aadA1) tetA, tetA (tetA) mcr1, mcr1 mcr 1 (mcr-1) blaTEM bla TEM (bla-TEM blaCTXM, blaCTXM CTX M, M bla-CTX-M 86 86% 88 88% 83 77 77% respectivamente 3338 38 33/3 (86% quinolonas (bla-CTX-M mcr- CTX30 30 (CTX-30 IMP10. IMP10 IMP 10 . (IMP-10) clados forma Broilers (Broilers humano 20 15 (15 (aac(3)-IV (aadA1 (tetA (mcr-1 8 7 333 33/ (86 CTX3 (CTX-3 IMP1 (IMP-10 2 (1 (aadA (mcr- (8 (CTX- (IMP-1 ( (mcr (CTX (IMP- (IMP

Abstract Silver nanoparticles are opted to have various applications in different fields ranging from traditional medicines to culinary items. It is toxic and most effective against bacteria, fungi viruses, parasites, parasite carrying vectors such as mosquitoes and their larvae and other eukaryotic microorganisms at low concentration without any side effects and toxicity to humans. In view of these data, the present research has been investigated by synthesizing silver nanoparticles using 1mM silver nitrate and aqueous extract of Passiflora foetida. The variation of nanoparticles in size and shape concerning the concentration of extract prepared were analysed. The formation of silver nanoparticles was confirmed by colour changing from yellowish green to reddish-brown implicating the surface plasmon resonance. Further, it was concluded by obtaining an absorbance peak at 420 nm using UV-Visible spectrophotometer analysis. FTIR analysis was used to identify the capping ligands, which included alkanes, aromatic groups and nitro compounds. The average grain size of ~12 nm to 14 nm with crystalline phase was revealed by X-ray Diffraction studies. The SEM images depicted the surface morphology with agglomeration; TEM studies showed the shape of nanoparticles as spherical and hexagonal with sizes ranging from 40 nm to 100 nm and EDAX analysis confirmed the presence of elemental silver as the principal constituent. The characterized silver nanoparticles were then tested for synergistic antibacterial effects with tetracycline, and the results show that they are more active against E. coli and S. aureus, but moderately effective against B. cereus and K. pneumoniae . It also had a strong larval and pupal toxic effects on the dengue vector, Aedes aegypti with the highest mortality. As a result, silver nanoparticles could be a viable alternative for a variety of applications. items bacteria viruses parasites humans data mM foetida analysed reddishbrown reddish brown resonance Further 42 UVVisible UV Visible ligands alkanes compounds 12 ~1 1 Xray X ray agglomeration 4 10 constituent tetracycline E S aureus B K vector mortality result ~

Resumo Os nanopartículos de prata são optados por ter várias aplicações em diferentes áreas que variam de medicamentos tradicionais a itens culinários. É tóxico e mais eficaz contra bactérias, vírus de fungos, parasitas, parasitas que transportam vetores como mosquitos e suas larvas e outros microorganismos eucarióticos em baixa concentração, sem efeitos colaterais e toxicidade para os seres humanos. Em vista desses dados, a presente pesquisa foi investigada sintetizando nanopartículas de prata usando nitrato de prata de 1 mm e extrato aquoso de Passiflora foetida. Foi analisada a variação de nanopartículas em tamanho e forma relativa à concentração de extrato preparado. A formação de nanopartículas de prata foi confirmada pela mudança de cor de verde amarelado para marrom-avermelhado, implicando a ressonância plasmônica da superfície. Além disso, foi concluído pela obtenção de um pico de absorvância a 420 nm usando análise do espectrofotômetro visível por UV. A análise do FTIR foi usada para identificar os ligantes de captura, que incluíam alcanes, grupos aromáticos e compostos nitro. O tamanho médio dos grãos de ~ 12 nm a 14 nm com fase cristalina foi revelado por estudos de difração de raios-X. As imagens SEM retratavam a morfologia da superfície com aglomeração; Os estudos de TEM mostraram a forma das nanopartículas como esféricas e hexagonais, com tamanhos variando de 40 nm a 100 nm e a análise EDAX confirmou a presença de prata elementar como constituinte principal. As nanopartículas de prata caracterizadas foram então testadas quanto a efeitos antibacterianos sinérgicos com tetraciclina, e os resultados mostram que são mais ativos contra E. coli e S. aureus, mas moderadamente eficazes contra B. cereus e K. pneumoniae. Ele também teve um forte efeito tóxico larval e pupal no vetor de dengue, Aedes aegypti com a maior mortalidade. Como resultado, as nanopartículas de prata podem ser uma alternativa viável para uma variedade de aplicações. culinários bactérias fungos humanos dados foetida preparado marromavermelhado, marromavermelhado marrom avermelhado, avermelhado marrom-avermelhado disso 42 UV captura alcanes nitro raiosX. raiosX raios X. X raios-X aglomeração hexagonais 4 10 principal tetraciclina E S aureus B K pneumoniae dengue mortalidade resultado