A novel multitargeted self-assembling peptide-siRNA complex for simultaneous inhibition of SARS-CoV-2-host cell interaction and replication.

Effective therapeutics are necessary for managing severe COVID-19 disease despite the availability of vaccines. Small interfering RNA (siRNA) can silence viral genes and restrict SARS-CoV-2 replication. Cell-penetrating peptides is a robust method for siRNA delivery, enhancing siRNA stability and targeting specific receptors. We developed a peptide HE25 that blocks SARS-CoV-2 replication by various mechanisms, including the binding of multiple receptors involved in the virus's internalization, such as ACE2, integrins and NRP1. HE25 not only acts as a vehicle to deliver the SARS-CoV-2 RNA-dependent RNA polymerase siRNA into cells but also facilitates their internalization through endocytosis. Once inside endosomes, the siRNA is released into the cytoplasm through the Histidine-proton sponge effect and the selective cleavage of HE25 by cathepsin B. These mechanisms effectively inhibited the replication of the ancestral SARS-CoV-2 and the Omicron variant BA.5 in vitro. When HE25 was administered in vivo, either by intravenous injection or inhalation, it accumulated in lungs, veins and arteries, endothelium, or bronchial structure depending on the route. Furthermore, the siRNA/HE25 complex caused gene silencing in lung cells in vitro. The SARS-CoV-2 siRNA/HE25 complex is a promising therapeutic for COVID-19, and a similar strategy can be employed to combat future emerging viral diseases.

The Role of the Fusarium oxysporum FTF2 Transcription Factor in Host Colonization and Virulence in Common Bean Plants (Phaseolus vulgaris L.).

The FTF (Fusarium Transcription Factor) gene family is composed of two members (FTF1 and FTF2) with high-sequence homology that encode transcription factors involved in the modulation of virulence in the F. oxysporum species complex (FOSC). While FTF1 is a multicopy gene exclusive of highly virulent strains of FOSC and is located in the accessory genome, FTF2 is a single-copy gene, located in the core genome, and well-conserved in all filamentous ascomycete fungi, except yeast. The involvement of FTF1 in the colonization of the vascular system and regulation of the expression of SIX effectors has been stablished. To address the role of FTF2, we generated and characterized mutants defective in FTF2 in a F. oxysporum f. sp. phaseoli weakly virulent strain and analyzed them together with the equivalent mutants formerly obtained in a highly virulent strain. The results obtained highlight a role for FTF2 as a negative regulator of the production of macroconidia and demonstrate that it is required for full virulence and the positive regulation of SIX effectors. In addition, gene expression analyses provided compelling evidence that FTF2 is involved in the regulation of hydrophobins likely required for plant colonization.

Vision-Based Module for Herding with a Sheepdog Robot.

Livestock farming is assisted more and more by technological solutions, such as robots. One of the main problems for shepherds is the control and care of livestock in areas difficult to access where grazing animals are attacked by predators such as the Iberian wolf in the northwest of the Iberian Peninsula. In this paper, we propose a system to automatically generate benchmarks of animal images of different species from iNaturalist API, which is coupled with a vision-based module that allows us to automatically detect predators and distinguish them from other animals. We tested multiple existing object detection models to determine the best one in terms of efficiency and speed, as it is conceived for real-time environments. YOLOv5m achieves the best performance as it can process 64 FPS, achieving an mAP (with IoU of 50%) of 99.49% for a dataset where wolves (predator) or dogs (prey) have to be detected and distinguished. This result meets the requirements of pasture-based livestock farms.

Primer reporte en Cuba de Anisakis spp. en muestras de pescado en conserva / First report in Cuba of Anisakis spp. in samples of canned fish

Introducción: Las especies de Anisakis representan uno de los grupos parasitarios patógenos transmitidos por alimentos más importantes en relación con sus capacidades zoonóticas y pueden producir una enfermedad emergente llamada anisakiosis. En Cuba, no existen reportes de la enfermedad como tampoco publicaciones sobre la detección del parásito en muestras de productos pesqueros. Objetivo: Identificar larvas de Anisakis spp. en muestras de productos pesqueros. Métodos: Se realizó un estudio para la búsqueda de larvas de Anisakis spp. en muestras de productos pesqueros en conserva recibidos en el Laboratorio de Parasitología de los alimentos del Instituto Nacional de Higiene, Epidemiología y Microbiología de Cuba. Se recibieron 96 muestras de dos marcas diferentes. A cada marca se le asignó las letras A o B para proteger la identidad del fabricante. Las larvas recuperadas fueron previamente aclaradas con lactofenol e identificadas morfológicamente por microscopía. Se utilizaron las claves de identificación para larvas de Anisakis spp. descritas en la literatura. Resultados: El 38,5 por ciento del total de muestras de pescado en conserva analizadas estuvieron parasitadas con larvas de Anisakis spp., con una frecuencia media de 1,2 larvas por cada submuestra. Las larvas fueron clasificadas de tercer estadio y de tipo I. Conclusiones: Por primera vez se identificó en Cuba larvas de Anisakis spp. en muestras de pescado en conserva y se proporcionó datos de interés a las autoridades sanitarias para el fortalecimiento de políticas públicas relacionadas con la calidad e inocuidad de los alimentos(AU)

Introduction: Anisakis species represent one of the most important foodborne pathogenic parasitic groups in relation to their zoonotic capabilities, and can produce an emerging disease called anisakiosis. In Cuba, there are no reports of the disease and there are no publications on the detection of the parasite in samples of fishery products. Objective: To identify Anisakis spp. larvae in samples of fishery products. Methods: A study was conducted to search for Anisakis spp. larvae in samples of canned fish products received at the Laboratory of Food Parasitology of the National Institute of Hygiene, Epidemiology and Microbiology of Cuba. Ninety-six samples of two different brands were received. Each brand was assigned letter A or B to protect the identity of the manufacturer. The larvae recovered were previously rinsed with lactophenol and identified morphologically by microscopy. The identification keys for Anisakis spp. larvae described in the literature were used. Results: 38.5 percent of the total canned fish samples analyzed were parasitized with Anisakis spp. larvae, with a mean frequency of 1.2 larvae per subsample. The larvae were classified as third instar and type I. Conclusions: For the first time in Cuba, Anisakis spp. larvae were identified in canned fish samples and provided data of interest to health authorities to strengthen public policies related to food quality and safety(AU)

In Planta Gene Expression Analysis and Colonization of Fusarium oxysporum.

In planta gene expression analysis and GFP-based confocal microscopy are two powerful techniques that may be coupled to assess the extent and dynamics of plant colonization by a fungal pathogen. Here we describe methods to prepare common bean plants for inoculation with a highly virulent strain of Fusarium oxysporum f. sp. phaseoli, quantify the extent of colonization by RT-qPCR, and visualize the colonized tissues by confocal microscopy.

Agrobacterium tumefaciens-Mediated Transformation of NHEJ Mutant Aspergillus nidulans Conidia: An Efficient Tool for Targeted Gene Recombination Using Selectable Nutritional Markers.

Protoplast transformation for the introduction of recombinant DNA into Aspergillus nidulans is technically demanding and dependant on the availability and batch variability of commercial enzyme preparations. Given the success of Agrobacterium tumefaciens-mediated transformation (ATMT) in diverse pathogenic fungi, we have adapted this method to facilitate transformation of A. nidulans. Using suitably engineered binary vectors, gene-targeted ATMT of A. nidulans non-homologous end-joining (NHEJ) mutant conidia has been carried out for the first time by complementation of a nutritional requirement (uridine/uracil auxotrophy). Site-specific integration in the ΔnkuA host genome occurred at high efficiency. Unlike other transformation techniques, however, cross-feeding of certain nutritional requirements from the bacterium to the fungus was found to occur, thus limiting the choice of auxotrophies available for ATMT. In complementation tests and also for comparative purposes, integration of recombinant cassettes at a specific locus could provide a means to reduce the influence of position effects (chromatin structure) on transgene expression. In this regard, targeted disruption of the wA locus permitted visual identification of transformants carrying site-specific integration events by conidial colour (white), even when auxotrophy selection was compromised due to cross-feeding. The protocol described offers an attractive alternative to the protoplast procedure for obtaining locus-targeted A. nidulans transformants.

Susceptibilidad antimicrobiana y serovariedades de Salmonella aisladas en carnes y productos cárnicos / Antimicrobial susceptibility and serovarieties of Salmonella isolated from meat and meat products

Introducción: La carne de ave y otros tipos de carnes provenientes de animales infectados son importantes vehículos de salmonelosis. Objetivo: Determinar la susceptibilidad a los antimicrobianos y las serovariedades de Salmonella aisladas en carnes y productos cárnicos. Material y Métodos: Se analizaron 172 aislados en el período enero de 2012 a marzo de 2020 en el laboratorio de Microbiología del Instituto Nacional de Higiene Epidemiología y Microbiología de Cuba. El serotipaje se determinó según la norma ISO/TR 6579-3:2014. La susceptibilidad antimicrobiana se realizó mediante el método de difusión con discos (Bauer-Kirby) de acuerdo con la metodología descrita en la normativa del Instituto de Normas Clínicas y de Laboratorio. Resultados: Las serovariedades identificadas con mayor frecuencia fueron: S. Enteritidis, S. Agona, S. Derby, S. Infantis y S. London, 18 (43,9 %) serovariedades identificadas en las carnes frescas también se encontraron en las procesadas. Los porcentajes más altos de resistencia antimicrobiana se hallaron frente al ácido nalidíxico, la tetraciclina y la ampicilina.S. Enteritidis y S. Typhimurium expresaron resistencia a un mayor número de antibióticos. Conclusiones: Los resultados sugieren que las carnes frescas constituyen una fuente de diseminación de Salmonella, incluso aquellas portadoras de resistencia a los antimicrobianos(AU)

Poultry and other types of meat from infected animals are important vehicles of salmonellosis. Objective: To determine the susceptibility to antimicrobial agents and serovarieties of Salmonella isolated from meat and meat products. Material and Methods: A total of 159 isolates were analyzed at the Cuban National Institute of Hygiene, Epidemiology and Microbiology during the period between January 2012 and March 2020. Serotypes were determined according to ISO/TR 6579-3: 2014. Antimicrobial susceptibility was determined by the Bauer-Kirby technique, according to the methodology described in the regulations of the Clinical and Laboratory Standards Institute. Results: The most frequent serovarieties identified were S. Enteritidis, S. Agona, S. Derby, S. Infantis and S. London. Also, 18 (43, 9 %) of serovarieties identified in fresh meat were found in processed meat. The highest percentages were related to antimicrobial resistance to nalidixic acid, tetracycline and ampicillin. S. Enteritidis and S. Typhimurium serotypes showed resistance to a greater number of antibiotics. Conclusions: The results suggest that fresh meats are an important source of Salmonella contamination, including those that are carriers of antimicrobial-resistant pathogens(AU)

Perfil del riesgo de brotes alimentarios por intoxicación estafilocócica en Cuba / Risk profile of staphylococcal food poisoning outbreaks in Cuba

Introducción: Staphylococcus ocasiona con frecuencia intoxicaciones alimentarias; en Cuba es una de las principales causas de brotes. Objetivo: analizar el perfil del riesgo de brotes alimentarios por Staphylococcus en Cuba. Métodos: Se realizó un estudio observacional analítico sobre los factores higiénicos y epidemiológicos asociados al riesgo de la intoxicación alimentaria por Staphylococcus en Cuba, en el Instituto Nacional de Higiene Epidemiología y Microbiología, durante el periodo de enero a noviembre de 2018. La información se obtuvo mediante revisión bibliográfica sistémica, análisis de datos del Laboratorio de Referencia Nacional en Microbiología Sanitaria y informes de brotes de la Dirección Nacional de Salud Ambiental. Se estimó el riesgo microbiológico mediante el programa semicuantitativo Ross-Sumner. Resultados: Se identificó como principal peligro Stahylococcus aureus, productores de enterotoxina A y la determinación de aislamientos resistentes a los antimicrobianos, como Stahylococcus aureus meticilina resistentes. Los alimentos más implicados fueron los productos de repostería y la inocuidad se afectó en mayor frecuencia por la contaminación cruzada en el 31,4 por ciento brotes y la inadecuada conservación en 46,1 por ciento. La predicción de personas que enferman por año en la población de interés fue de 6 260. Conclusiones: El riesgo de la ocurrencia de brotes por Staphylococcus se estimó como alto; la vigilancia se recomienda en alimentos que requieren manipulación directa y no se aplica tratamiento térmico antes del consumo, con gestiones de riesgo dirigidas a productores y consumidores(AU)

Introduction: Staphylococcus is a frequent cause of food poisoning. In Cuba it is one of the main causes of outbreaks. Objective: Analyze the risk profile of staphylococcal food poisoning outbreaks in Cuba. Methods: An observational analytical study was conducted about hygienic and epidemiological factors associated to the risk for staphylococcal food poisoning in Cuba. The study was carried out at the National Institute of Hygiene, Epidemiology and Microbiology from January to November 2018. The information was obtained from a systematic bibliographic review, analysis of data from the Medical Microbiology National Reference Laboratory, and outbreak reports from the National Environmental Health Division. Microbiological risk was estimated with Ross-Sumner semi-quantitative software. Results: The main hazard identified was Staphylococcus aureus, enterotoxin A producers and determination of antimicrobial resistant isolates like methicillin-resistant Staphylococcus aureus. The food items most commonly involved were sweets, and safety was most frequently affected by cross-contamination in 31.4 percent of the outbreaks and inadequate preservation in 46.1 percent. Prediction of members of the population of interest becoming ill every year was 6 260. Conclusions: Risk for the occurrence of staphylococcal food poisoning outbreaks was considered to be high; surveillance is recommended of food items requiring direct manipulation and appropriate thermal treatment is not applied before consumption, with risk management actions aimed at manufacturers and consumers(AU)

Specific tissue proteins 1 and 6 are involved in root biology during normal development and under symbiotic and pathogenic interactions in Medicago truncatula.

MAIN CONCLUSION: ST1 and ST6 are possibly involved in primary and lateral root and symbiotic nodule development, but only ST6 participates in the interaction with hemibiotrophic fungi. Specific tissue (ST) proteins have been shown to be involved in several processes related to plant nutritional status, development, and responses to biotic agents. In particular, ST1 and ST6 are mainly expressed in roots throughout plant development. Here, we analyze where and how the expression of the genes encoding both proteins are modulated in the legume model plant Medicago truncatula in response to the plant developmental program, nodulation induced by a beneficial nitrogen-fixing bacterium (Sinorhizobium meliloti) and the defense response triggered by a pathogenic hemibiotrophic fungus (Fusarium oxysporum). Gene expression results show that ST1 and ST6 participate in the vasculature development of both primary and lateral roots, although only ST6 is related to meristem activity. ST1 and ST6 clearly display different roles in the biotic interactions analyzed, where ST1 is activated in response to a N2-fixing bacterium and ST6 is up-regulated after inoculation with F. oxysporum. The role of ST1 and ST6 in the nodulation process may be related to nodule organogenesis rather than to the establishment of the interaction itself, and an increase in ST6 correlates with the activation of the salicylic acid signaling pathway during the infection and colonization processes. These results further support the role of ST6 in response to hemibiotrophic fungi. This research contributes to the understanding of the complex network that controls root biology and strengthens the idea that ST proteins are involved in several processes such as primary and lateral root development, nodule organogenesis, and the plant-microbe interaction.

Antimicrobial Resistance in Bacteria Isolated from Foods in Cuba.

INTRODUCTION Antimicrobial drug resistance constitutes a health risk of increasing concern worldwide. One of the most common av-enues for the acquisition of clinically-relevant antimicrobial resistance can be traced back to the food supply, where resistance is acquired through the ingestion of antimicrobial resistant microorganisms pres-ent in food. Antimicrobial resistance constitutes a health risk, leading to production losses and negative consequences for livelihood and food safety. OBJECTIVE Determine whether resistant bacteria are present in foods in Cuba. METHODS A descriptive observational study was conducted in theMicrobiology Laboratory of Cuba's National Institute of Hygiene, Epi-demiology and Microbiology from September 2004 through Decem-ber 2018. Researchers analyzed 1178 bacterial isolates from food samples. The isolates were identifi ed as Escherichia coli, Salmonella, Vibrio cholerae and coagulase-positive Staphylococcus. The antimi-crobial susceptibility study was performed using the Bauer-Kirby disk diffusion method, following procedures outlined by the Clinical and Laboratory Standards Institute. The data were analyzed using WHO-NET version 5.6. RESULTS Of the total isolates, 62.1% were resistant to at least one antibiotic. Within each group, >50% of isolates showed some type of resistance. E. coli and V. cholerae exceeded 50% resistance to tetracycline and ampicillin, respectively. Staphylococcus showed the highest resistance to penicillin, and Salmonella to tetracycline, nali-dixic acid and ampicillin. The highest percentages of non-susceptible microorganisms were identifi ed in meats and meat products. CONCLUSIONS These results serve as an alert to the dangers of acquiring antibiotic-resistant bacteria from food and demonstrate the need to establish a surveillance system and institute measures bacte-rial control in food products.KEYWORDS Microbial drug resistance, bacteria, food, foodborne disease, Cuba.

Fibroadenoma gigante juvenil de mama: presentación de un caso clínico / Juvenile giant fibroadenoma of the breast: presentation of a clinical case

RESUMEN El fibroadenoma gigante juvenil es un tumor de mama benigno y una variante rara de los fibroadenomas. La presentación clínica suele ser una masa tumoral unilateral, de crecimiento rápido e indolora. En este artículo presentamos el caso de un fibroadenoma gigante juvenil de 12 cm de diámetro en la mama de una niña de 13 años. Se realiza estudio radiológico e histológico de la lesión siendo categorizada como un fibroadenoma gigante juvenil por lo que se realiza tumorectomía completa con remodelación mamaria posterior. A los dos meses de seguimiento, la paciente se encuentra sin signos de recidiva, con buena situación general y a la espera de cirugía de remodelación mamaria.

ABSTRACT Juvenile giant fibroadenoma is a benign breast tumor and a rare variant of fibroadenomas. The clinical presentation is usually a painless, fast growing, unilateral tumor mass. In this article we present the case of a giant juvenile fibroadenoma of 12 cm in diameter in the breast of a 13-year-old girl. A radiological and histological study of the lesion was carried out and it was categorized as a juvenile giant fibroadenoma, so a complete lumpectomy with posterior breast remodeling was performed. After two months of follow-up, the patient is without signs of recurrence, in good general condition and waiting for the breast remodeling surgery.

Resistencia antimicrobiana en bacterias aisladas de pescados y mariscos / Antimicrobial resistance in bacteria isolated in fish and shellfish

RESUMEN Introducción: El alarmante incremento de la resistencia bacteriana frente a los antibióticos constituye uno de los mayores problemas en Salud Pública. Las bacterias de ambientes acuáticos pueden transferir genes de resistencia antimicrobiana a otras bacterias, incluyendo las patógenas para el hombre, lo cual constituye un riesgo. Objetivo: Determinar la resistencia a los antimicrobianos en aislamientos bacterianos de pescados y mariscos. Materiales y métodos: Se analizaron 154 aislamientos bacterianos en pescados y mariscos en el Laboratorio de Microbiología del Instituto Nacional de Higiene Epidemiología y Microbiología. La susceptibilidad antimicrobiana, se determinó mediante el método de difusión con discos (Bauer-Kirby), según la normativa del Clinical and Laboratory Standards Institute. El análisis de los resultados se realizó mediante el programa WHONET 5.6. Resultados: Se Identificó resistencia en dos cepas de Salmonella y seis de E. coli, la resistencia se expresó con mayor frecuencia para para la ampicilina y la tetraciclina. En Staphylococcus se determinó un patrón de multirresistencia para el cloranfenicol, la eritromicina y la tetraciclina. Vibrio cholerae fue el género con mayor aislamiento en pescados y mariscos, las cepas resistentes fueron más frecuentes en ostiones y pescados de aguas dulces. Conclusiones: Se determinaron los mayores porcentajes de resistencia para la ampicilina y la tetraciclina en el total de los microorganismos estudiados. El ostión se consideró el producto de mayor riesgo de diseminación de bacterias resistentes a los antimicrobianos.

ABSTRACT Introduction: The alarming increase of antibiotics resistance in bacteria is one of the greatest problems in Public Health. Bacteria in aquatic environments can transfer antimicrobial resistance genes to other bacteria, including pathogenic bacteria for humans, which is a health risk. Objective: To determine the antimicrobial resistance in bacterial isolates from fish and shellfish. Materials and methods: A total of 154 bacterial isolates were analyzed in fish and shellfish in the microbiology laboratory of the National Institute of Hygiene, Epidemiology and Microbiology. Antimicrobial susceptibility was determined by the Kirby- Bauer disc diffusion method according to the regulations of the Clinical and Laboratory Standards Institute. The analysis of the results was carried out by the WHONET 5.6 program. Results: Resistance was identified in two strains of Salmonella and six ones of Escherichia coli, the resistance to ampicillin and tetracycline was higher. A multiresistance pattern was identified in multiresistant staphylococcus to chloramphenicol, erythromycin and tetracycline. Vibrio cholerae was the most frequent genus found in fish and shellfish; resistant strains were more frequent in oysters and freshwater fish. Conclusions: The highest percentages of resistance were determined for ampicillin and tetracycline in the total of microorganisms studied. The oyster was considered the greatest risk product of dissemination of antimicrobial-resistant bacteria.

Ausencia de actividad antimicrobiana de un extracto acuoso liofilizado de Capraria biflora L. (esclaviosa) / Absence of antimicrobial activity in a lyophilized aqueous extract from Capraria biflora L. (goatweed)

Introducción: la especie Capraria biflora L. perteneciente a la familia Scrophulariaceae, presenta una larga historia en la medicina tradicional por tener gran aplicación en varios tratamientos: cuyas causas podrían estar relacionados con infecciones antimicrobianas. Objetivo: Determinar la actividad antimicrobiana del extracto acuoso liofilizado de la especie Capraria biflora L. frente a una batería mínima de microorganismos de interés clínico. Métodos: la obtención del extracto se realizó por secado en horno, pulverización en un mortero, extracción por decocción y posterior liofilización. La actividad antibacteriana del extracto acuoso se evaluó in vitro, en dos concentraciones (10 y 50 mg/mL), mediante el sistema de ensayo de difusión en agar, con una batería mínima de cepas de microorganismos compuesta por cuatro bacterias Gram negativas: Klebsiella neumoneae (ATCC 10031), Salmonella tiphymurium (ATCC 14028), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), una bacteria Gram positiva: Staphylococcus aureus (ATCC 6538), y dos levaduras Cándida albicans (ATCC 10231) y Saccharomyces cerevisiae (BCTP 20/agosto/2008). Resultados: el extracto acuoso de las hojas de Capraria biflora L no presenta actividad antimicrobiana, a las concentraciones estudiadas, frente a las cepas de microorganismos de interés clínico utilizadas en el ensayo. Conclusiones: se reporta por primera vez la evaluación de la actividad antimicrobiana del extracto acuoso liofilizado de las hojas de Capraria biflora L. frente a cepas de bacterias y hongos de interés clínico a las concentraciones de 10 y 50 mg/mL. No se encontró un efecto inhibitorio y se recomienda realizar otros estudios de actividad antimicrobiana que inserten la temática de la virulencia microbiana, encaminados a evaluar el posible efecto bacteriostático que pueda presentar este extracto.

Introduction: the species Capraria biflora L., from the Scrophulariaceae family, has a long history of use in traditional medicine for the treatment of various disorders whose causes could be related to microbial infections. Objective: determine the antimicrobial activity of lyophilized aqueous extract from the species Capraria biflora L. against a minimum set of microorganisms of clinical interest. Methods: the extract was obtained by oven drying, pulverization in a mortar, decoction and lyophilization. In vitro antibacterial activity of the aqueous extract was evaluated at two concentrations (10 and 50 mg/ml) by the agar diffusion test with a minimum set of microorganism strains composed of four gram-negative bacteria: Klebsiella neumoneae (ATCC 10031), Salmonella tiphymurium (ATCC 14028), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), one gram-positive bacterium: Staphylococcus aureus (ATCC 6538), and two yeasts: Candida albicans (ATCC 10231) and Saccharomyces cerevisiae (BCTP 20/08/2008). Results: at the study concentrations, the aqueous extract of Capraria biflora L. leaves does not display antimicrobial activity against the strains of microorganisms of clinical interest evaluated in the assay. Conclusions: this is the first evaluation of the antimicrobial activity of lyophilized aqueous extract from leaves of Capraria biflora L. against strains of bacteria and fungi of clinical interest at concentrations of 10 and 50 mg/ml. No inhibitory effect was found. It is recommended to conduct further antimicrobial activity studies incorporating the topic of microbial virulence, aimed at evaluating the potential bacteriostatic effect of the study extract.

Characterization of chronic clinical manifestations in patients with chikungunya fever / Caracterización de las manifestaciones clínicas crónicas en pacientes con fiebre chikunguña

Background: In Mexico there are no studies concerning the chronicity of clinical manifestations of chikungunya fever (CHIK). Objective: To describe the characterization of chronic clinical manifestations in patients with chikungunya fever from the Unidad de Medicina Familiar No.9 (UMF No. 9). Methods: A longitudinal, ambispective study was performed in a cohort of 116 patients with positive serology of CHIK, confirmed by RTPCR or IGM test performed at UMF No. 9 of Acapulco, Guerrero, from December 2014 to January 2017. The population was obtained from the epidemiological surveillance platform. The source of information was the epidemiological study of cases for vector-borne diseases. With measurements at 3, 4, 5, 6, 12 and 18 months of the onset of the acute phase, data were captured and analyzed in the statistical package SPSS, version 23. Results: 62% of patients presented chronicity at 3 months; arthralgia was the most frequent chronic symptom with 88.8%; female gender and the age group of 20-59 years were the most affected. Conclusions: Even though more than half of our population presented CHIK chronicity, this was lower than the reports from international literature; the main symptoms reported during the chronic phase were arthralgias and fatigue. The knowledge of CHIK symptoms behavior provides standards to generate integral management guides with a holistic and multidisciplinary approach.

Introducción: En México no hay estudios acerca de la cronicidad de las manifestaciones clínicas de la fiebre chikunguña (CHIK). Objetivo: Describir la caracterización de las manifestaciones clínicas crónicas en pacientes con CHIK de la Unidad de Medicina Familiar No. 9 (UMF No. 9). Métodos: Se realizó un estudio longitudinal ambispectivo, en una cohorte de 116 pacientes con serología positiva para CHIK, confirmada por prueba RTPCR o IGM realizada en la UMF No. 9 de Acapulco, Guerrero, de diciembre de 2014 a enero de 2017. La población se obtuvo de la plataforma de vigilancia epidemiológica de los sistemas especiales. La fuente de información fue el estudio epidemiológico de casos para enfermedades transmitidas por vector. Con mediciones a los 3, 4, 5, 6, 12 y 18 meses de iniciada la fase aguda, los datos se capturaron y analizaron en el paquete estadístico SPSS, versión 23. Resultados: El 62% de los pacientes presentó cronicidad a los 3 meses; la artralgia fue el síntoma crónico más frecuente con 88.8%; el género femenino y el grupo de edad de 20-59 fueron los más afectados. Conclusiones: Más de la mitad de nuestra población presentó cronicidad por CHIK, frecuencia menor a la reportada en la literatura internacional; los principales síntomas referidos durante la fase crónica fueron artralgias y fatiga. El conocimiento del comportamiento de los síntomas de la CHIK nos da la pauta para generar guías de manejo integral con un enfoque holístico y multidisciplinario.

The FTF gene family regulates virulence and expression of SIX effectors in Fusarium oxysporum.

The FTF (Fusarium transcription factor) gene family comprises a single copy gene, FTF2, which is present in all the filamentous ascomycetes analysed, and several copies of a close relative, FTF1, which is exclusive to Fusarium oxysporum. An RNA-mediated gene silencing system was developed to target mRNA produced by all the FTF genes, and tested in two formae speciales: F. oxysporum f. sp. phaseoli (whose host is common bean) and F. oxysporum f. sp. lycopersici (whose host is tomato). Quantification of the mRNA levels showed knockdown of FTF1 and FTF2 in randomly isolated transformants of both formae speciales. The attenuation of FTF expression resulted in a marked reduction in virulence, a reduced expression of several SIX (Secreted In Xylem) genes, the best studied family of effectors in F. oxysporum, and lower levels of SGE1 (Six Gene Expression 1) mRNA, the presumptive regulator of SIX expression. Moreover, the knockdown mutants showed a pattern of colonization of the host plant similar to that displayed by strains devoid of FTF1 copies (weakly virulent strains). Gene knockout of FTF2 also resulted in a reduction in virulence, but to a lesser extent. These results demonstrate the role of the FTF gene expansion, mostly the FTF1 paralogues, as a regulator of virulence in F. oxysporum and suggest that the control of effector expression is the mechanism involved.

Gene expression patterns and dynamics of the colonization of common bean (Phaseolus vulgaris L.) by highly virulent and weakly virulent strains of Fusarium oxysporum.

The dynamics of root and hypocotyl colonization, and the gene expression patterns of several fungal virulence factors and plant defense factors have been analyzed and compared in the interaction of two Fusarium oxysporum f. sp. phaseoli strains displaying clear differences in virulence, with a susceptible common bean cultivar. The growth of the two strains on the root surface and the colonization of the root was quantitatively similar although the highly virulent (HV) strain was more efficient reaching the central root cylinder. The main differences between both strains were found in the temporal and spatial dynamics of crown root and hypocotyl colonization. The increase of fungal biomass in the crown root was considerably larger for the HV strain, which, after an initial stage of global colonization of both the vascular cylinder and the parenchymal cells, restricted its growth to the newly differentiated xylem vessels. The weakly virulent (WV) strain was a much slower and less efficient colonizer of the xylem vessels, showing also growth in the intercellular spaces of the parenchyma. Most of the virulence genes analyzed showed similar expression patterns in both strains, except SIX1, SIX6 and the gene encoding the transcription factor FTF1, which were highly upregulated in root crown and hypocotyl. The response induced in the infected plant showed interesting differences for both strains. The WV strain induced an early and strong transcription of the PR1 gene, involved in SAR response, while the HV strain preferentially induced the early expression of the ethylene responsive factor ERF2.

Calidad microbiológica de las hortalizas y factores asociados a la contaminación en áreas de cultivo en La Habana / Microbiological quality of vegetables and factors associated with contamination in growing areas in Havana

Introducción: en los últimos años se han incrementado las enfermedades transmitidas por frutas y hortalizas, por lo que es importante evaluar los factores que afectan la inocuidad de estos productos. Objetivo: determinar la calidad microbiológica de hortalizas y los factores asociados a la contaminación en áreas de cultivo en La Habana. Materiales y métodos: se estudiaron 100 muestras de vegetales de 26 áreas y el agua de regadío de cada plantación, en el período de enero del 2009 a diciembre del 2011, en el Instituto de Nutrición e Higiene de los Alimentos. La determinación de parásitos se realizó según procedimiento descrito en el Manual de Análisis Bacteriológico FDA / CFSAN 2001, para el estudio bacteriológico de las hortalizas y el agua se emplearon las normas vigentes en el país. Resultados: se determinó la presencia de parásitos en 6 por ciento de los vegetales y Escherichia coli en 18,0 por ciento, con mayor frecuencia en lechuga, berro y espinaca. Se aislaron bacterias patógenas, Salmonella Weltevreden en cebollino y potencialmente patógena Listeria spp. en acelga. El 53,8 por ciento de las muestras de agua no tuvo una calidad microbiológica aceptable. El no cercado de las áreas de cultivo, la presencia de animales en el campo y el uso de agua contaminada, fueron los factores que más se observaron, encontrándose asociación estadística entre estos y la contaminación las hortalizas. Conclusiones: se detectó la presencia de parásitos, bacterias patógenas y potencialmente patógenas en las hortalizas estudiadas, lo que estuvo asociado principalmente al uso de agua de regadío no tratada, la presencia de animales en el campo y el no cercado de las áreas de cultivo(AU)

Introduction: over the last several years, there has been an increased a foodborne illness linked fruits and vegetables; therefone is important to evaluate factors affecting the safety of vegetal produce. Objective: to determine the microbiological quality of vegetables and factors associated with contamination in growing areas in Havana. Materials and Methods: 100 samples of vegetables from 26 areas were studied and irrigation water from each orchard, in the period January 2009 to December 2011 at the Institute of Nutrition and Food Hygiene. The determination of parasitic was performed according to the FDA Bacteriological Analytical Manual / CFSAN 2001, bacteriological study of vegetables and water was carried by the current standards in the country. Results: parasites was observed in 6% of vegetables and Escherichia coli in 18.0 percent, more frequently in lettuce, watercress and spinach. Were isolated pathogenic bacteria, Salmonella Weltevreden in chives and potentially pathogenic Listeria spp. in chard. Water samples 53.8 percent, did not have an acceptable microbiological quality. The fencing is not growing areas, the presence of animals in the field and the use of contaminated water were most frequently factors to observed, statistical association was found between these factors and vegetables contamination. Conclusions: it was detected the presence of the parasites, pathogen bacteria and potentially pathogen bacteria studied, and it was related to non-treated water for irrigation, the presence of animals in the soil, and the non-closed of the cultivate areas(AU)

Calidad microbiológica de las hortalizas y factores asociados a la contaminación en áreas de cultivo en La Habana / Microbiological quality of vegetables and factors associated with contamination in growing areas in Havana

Introducción: en los últimos años se han incrementado las enfermedades transmitidas por frutas y hortalizas, por lo que es importante evaluar los factores que afectan la inocuidad de estos productos. Objetivo: determinar la calidad microbiológica de hortalizas y los factores asociados a la contaminación en áreas de cultivo en La Habana. Materiales y métodos: se estudiaron 100 muestras de vegetales de 26 áreas y el agua de regadío de cada plantación, en el período de enero del 2009 a diciembre del 2011, en el Instituto de Nutrición e Higiene de los Alimentos. La determinación de parásitos se realizó según procedimiento descrito en el Manual de Análisis Bacteriológico FDA / CFSAN 2001, para el estudio bacteriológico de las hortalizas y el agua se emplearon las normas vigentes en el país. Resultados: se determinó la presencia de parásitos en 6% de los vegetales y Escherichia coli en 18,0%, con mayor frecuencia en lechuga, berro y espinaca. Se aislaron bacterias patógenas, Salmonella Weltevreden en cebollino y potencialmente patógena Listeria spp. en acelga. El 53,8% de las muestras de agua no tuvo una calidad microbiológica aceptable. El no cercado de las áreas de cultivo, la presencia de animales en el campo y el uso de agua contaminada, fueron los factores que más se observaron, encontrándose asociación estadística entre estos y la contaminación las hortalizas. Conclusiones: se detectó la presencia de parásitos, bacterias patógenas y potencialmente patógenas en las hortalizas estudiadas, lo que estuvo asociado principalmente al uso de agua de regadío no tratada, la presencia de animales en el campo y el no cercado de las áreas de cultivo.

Introduction: over the last several years, there has been an increased a foodborne illness linked fruits and vegetables; therefone is important to evaluate factors affecting the safety of vegetal produce. Objective: to determine the microbiological quality of vegetables and factors associated with contamination in growing areas in Havana. Materials and Methods: 100 samples of vegetables from 26 areas were studied and irrigation water from each orchard, in the period January 2009 to December 2011 at the Institute of Nutrition and Food Hygiene. The determination of parasitic was performed according to the FDA Bacteriological Analytical Manual / CFSAN 2001, bacteriological study of vegetables and water was carried by the current standards in the country. Results: parasites was observed in 6% of vegetables and Escherichia coli in 18.0%, more frequently in lettuce, watercress and spinach. Were isolated pathogenic bacteria, Salmonella Weltevreden in chives and potentially pathogenic Listeria spp. in chard. Water samples 53.8%, did not have an acceptable microbiological quality. The fencing is not growing areas, the presence of animals in the field and the use of contaminated water were most frequently factors to observed, statistical association was found between these factors and vegetables contamination. Conclusions: it was detected the presence of the parasites, pathogen bacteria and potentially pathogen bacteria studied, and it was related to non-treated water for irrigation, the presence of animals in the soil, and the non-closed of the cultivate areas.

Factores epidemiológicos de interés en brotes de enfermedades transmitidas por alimentos en La Habana / Epidemiological factors of interest in outbreaks of food-borne diseases in Havana

Objetivo: determinar los factores de interés epidemiológicos relacionados con la ocurrencia de brotes de enfermedades transmitidas por alimentos en La Habana en el período 2006-2010. Métodos: se realizó un estudio descriptivo retrospectivo de 130 brotes de enfermedades trasmitidas por alimentos, analizados en el Instituto de Nutrición e Higiene de los Alimentos en el período de enero del 2006 a diciembre del 2010. Los datos fueron obtenidos de los registros del laboratorio de Microbiología de los Alimentos de dicha institución y de las encuestas epidemiológicas del Programa Nacional de Vigilancia de estas enfermedades, del Ministerio de Salud Pública. Resultados: los lugares de ocurrencia de brotes más reportados fueron los comedores obreros (43,4 por ciento), ocasionados con mayor frecuencia por Clostridium perfringens, y los brotes familiares (32,8 por ciento) fueron producidos por Staphylococcus coagulasa positiva y Salmonella. Las carnes y productos cárnicos fueron los alimentos más implicados (52,0 por ciento), en la mayoría procedentes de comedores obreros y de escuelas. Los brotes familiares se relacionaron principalmente con el consumo de productos de repostería. El 89,2 por ciento y el 71,0 por ciento de los alimentos no fueron aceptables, según los indicadores de calidad sanitaria coliformes totales y coliformes fecales, respectivamente. El insuficiente tiempo, la temperatura durante la cocción y el calentamiento o recalentamiento del alimento, la inadecuada conservación en frío o caliente y la contaminación cruzada fueron los factores contribuyentes que más incidieron en la ocurrencia de brotes. Conclusiones: la calidad microbiológica inaceptable de los alimentos estudiados denota el incumplimiento de las buenas prácticas en la elaboración y conservación de los alimentos, lo que propicia la contaminación y la multiplicación de bacterias patógena(AU)

Objective: determine the epidemiological factors of interest related to the occurrence of outbreaks of food-borne diseases in Havana from 2006 to 2010. Methods: a retrospective descriptive study was conducted of 130 outbreaks of food-borne diseases analyzed at the Institute of Nutrition and Food Hygiene from January 2006 to December 2010. The data were obtained from records kept at the Food Microbiology laboratory of the institution and from epidemiological surveys contained in the National Surveillance Program of the Ministry of Public Health. Results: the places where most outbreaks were reported were staff cafeterias at workplaces (43.4 percent ), mostly caused by Clostridiumperfringens, and households (32.8 percent), caused by coagulase positive Staphylococcus and Salmonella. The foods most commonly reported were meat and meat products (52.0 percent), mostly from cafeterias at workplaces and schools. Household outbreaks were mainly related to the intake of sweets. 89.2 percent and 71.0 percent of the foods were not acceptable according to health indicators for total and fecal coliforms, respectively. Insufficient cooking time, cooking temperature, heating or reheating, inadequate cold or hot preservation and cross-contamination were the main contributing factors leading to the occurrence of outbreaks. Conclusions: the unacceptable microbiological quality of the foods studied reveals non-compliance with best practices in food preparation and preservation, paving the way for contamination and multiplication of pathogenic bacteria(AU)

Factores epidemiológicos de interés en brotes de enfermedades transmitidas por alimentos en La Habana / Epidemiological factors of interest in outbreaks of food-borne diseases in Havana

Objetivo: determinar los factores de interés epidemiológicos relacionados con la ocurrencia de brotes de enfermedades transmitidas por alimentos en La Habana en el período 2006-2010. Métodos: se realizó un estudio descriptivo retrospectivo de 130 brotes de enfermedades trasmitidas por alimentos, analizados en el Instituto de Nutrición e Higiene de los Alimentos en el período de enero del 2006 a diciembre del 2010. Los datos fueron obtenidos de los registros del laboratorio de Microbiología de los Alimentos de dicha institución y de las encuestas epidemiológicas del Programa Nacional de Vigilancia de estas enfermedades, del Ministerio de Salud Pública. Resultados: los lugares de ocurrencia de brotes más reportados fueron los comedores obreros (43,4 por ciento), ocasionados con mayor frecuencia por Clostridium perfringens, y los brotes familiares (32,8 por ciento) fueron producidos por Staphylococcus coagulasa positiva y Salmonella. Las carnes y productos cárnicos fueron los alimentos más implicados (52,0 por ciento), en la mayoría procedentes de comedores obreros y de escuelas. Los brotes familiares se relacionaron principalmente con el consumo de productos de repostería. El 89,2 por ciento y el 71,0 por ciento de los alimentos no fueron aceptables, según los indicadores de calidad sanitaria coliformes totales y coliformes fecales, respectivamente. El insuficiente tiempo, la temperatura durante la cocción y el calentamiento o recalentamiento del alimento, la inadecuada conservación en frío o caliente y la contaminación cruzada fueron los factores contribuyentes que más incidieron en la ocurrencia de brotes. Conclusiones: la calidad microbiológica inaceptable de los alimentos estudiados denota el incumplimiento de las buenas prácticas en la elaboración y conservación de los alimentos, lo que propicia la contaminación y la multiplicación de bacterias patógena

Objective: determine the epidemiological factors of interest related to the occurrence of outbreaks of food-borne diseases in Havana from 2006 to 2010. Methods: a retrospective descriptive study was conducted of 130 outbreaks of food-borne diseases analyzed at the Institute of Nutrition and Food Hygiene from January 2006 to December 2010. The data were obtained from records kept at the Food Microbiology laboratory of the institution and from epidemiological surveys contained in the National Surveillance Program of the Ministry of Public Health. Results: the places where most outbreaks were reported were staff cafeterias at workplaces (43.4 percent ), mostly caused by Clostridiumperfringens, and households (32.8 percent), caused by coagulase positive Staphylococcus and Salmonella. The foods most commonly reported were meat and meat products (52.0 percent), mostly from cafeterias at workplaces and schools. Household outbreaks were mainly related to the intake of sweets. 89.2 percent and 71.0 percent of the foods were not acceptable according to health indicators for total and fecal coliforms, respectively. Insufficient cooking time, cooking temperature, heating or reheating, inadequate cold or hot preservation and cross-contamination were the main contributing factors leading to the occurrence of outbreaks. Conclusions: the unacceptable microbiological quality of the foods studied reveals non-compliance with best practices in food preparation and preservation, paving the way for contamination and multiplication of pathogenic bacteria