Resumen La mastitis bovina es una enfermedad que afecta a las ganaderías de pequeños y medianos productores de los cantones Cayambe y Pedro Moncayo, Provincia de Pichincha-Ecuador. El tratamiento de esta enfermedad resulta complicado debido a la variedad de microorganismos que la provocan. El presente estudio se enfocó en la determinación molecular por medio de reacción en cadena de la polimerasa (PCR) de los agentes etiológicos de la mastitis. Esta técnica presenta múltiples ventajas al reconocer familia, género y especie de microorganismos, y es un método capaz de detectar genes de resistencia de antibióticos, lo que resulta importante al momento de diagnosticar y tratar enfermedades. El objetivo de esta investigación se centró en la identificación de bacterias causantes de la mastitis bovina, utilizando pruebas bioquímicas y moleculares. Las pruebas bioquímicas como tinción Gram, Catalasa, Coagulasa, y Agar Manitol Sal fueron eficientes para obtener cepas puras y determinar el género de algunas bacterias. Se utilizaron primers específicos (RNA16S) para la identificación molecular de 9 agentes etiológicos causantes de la enfermedad en las unidades productivas. Los microorganismos encontrados fueron; Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, la mayoría presentes en mastitis clínica. Para detectar genes de resistencia se utilizaron primers específicos, de los cuales 7 muestras presentaron el gen para resistencia a blaTEM (β-lactámicos) y 6 muestras presentaron el gen para resistencia a tetA (tetraciclinas). Se identificó multirresistencia en las especies: Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

Abstract Bovine mastitis is a disease that affects the farms of small and medium producers in the cantons of Cayambe and Pedro Moncayo, Pichincha Province-Ecuador. Treating this disease is not easy due to the different microorganisms that cause it. This study focused on the molecular determination by means of polymerase chain reaction (PCR) of the etiological agents of mastitis, having multiple advantages when recognizing family, gender and species of microorganisms. It is a method capable of detecting resistance genes of antibiotics, an important analysis when diagnosing and treating diseases. The aim of this research is to identify bacteria causing bovine mastitis by using biochemical and molecular tests. Biochemical tests such as: Gram staining, Catalase, Coagulase, and Mannitol Salt Agar were efficient to obtain pure strains and determine the gender of some bacteria. Specific primers (RNA16S) were used for the molecular identification of 9 etiological agents causing the disease in the productive units. The microorganisms found were Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, mostly present in clinic mastitis. To detect resistance genes, specific primers were used, of which 7 samples presented the gene for resistance to blaTEM (β-lactam) and 6 samples presented the gene for resistance to tetA (tetracyclines). Multi-resistance was identified in the species Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

Abstract The development of living beings depends on the health and chemical quality of the available water. In recent years, the importance of the environment in the spread of resistant and multi-resistant bacteria to antibiotics has been observed, a situation that has placed the problem of antibiotic resistance on a broader level and which is being studied from the perspective “One Health”. The objective of this work was to determine the presence of multi-resistant bacteria to antibiotics in irrigation water samples from the Chibunga river. Fourteen water samples of 100 mL each were collected in sterile containers and aseptically, were kept under refrigeration until analysis in the laboratory. Each sample was seeded by the surface seeding method on blood, cystine electrolyte deficient, MacConkey, Salmonella-Shigella and thiosulfate citrate bile sucrose agars, incubating in all cases at a temperature of 37 °C for a maximum time of 48 hours. The identification of bacterial colonies was carried out through physiological and biochemical tests according to the identification schemes indicated by MacFaddin. The resistance/sensitivity to antibiotics of the identified colonies was carried out by the disk diffusion method of Kirby Bauer. Two species of multi-resistant bacteria were identified, including strains of Morganella morgani and Plesiomonas shigelloides resistant to antibiotics of clinical use such as ceftazidime, aztreonam, ciprofloxacin, nalidixic acid and trimethoprim-sulfamethoxazole. The results show that the waters of Chibunga river harbor bacteria with multi-resistance to antimicrobials, representing a contamination risk of agricultural products harvested in its vicinity, as well as for the people who use these waters.

Resumen El bienestar y desarrollo de los seres vivos depende de la calidad sanitaria y química del agua disponible. En los últimos años se ha venido observando la importancia del ambiente en la diseminación de bacterias resistentes y multirresistentes a los antibióticos, situación que ha colocado la problemática de la resistencia en un plano más amplio y que se está estudiando desde la perspectiva “One Health”. En este sentido, el objetivo del presente trabajo fue determinar la presencia de bacterias multirresistentes a los antibióticos en muestras de agua de riego provenientes del río Chibunga. Se recolectaron 14 muestras de agua de un volumen de 100 mL cada una, en envases estériles y de manera aséptica, las cuales fueron conservadas bajo refrigeración hasta su análisis en el laboratorio. Cada muestra fue sembrada por el método de siembra en superficie en los agares cistina electrolito deficiente, MacConkey, Salmonella-Shigella, tiosulfato citrato bilis sacarosa y agar sangre, incubándose en todos los casos a una temperatura de 37 °C durante un tiempo máximo de 48 horas. La identificación de las colonias bacterianas se realizó mediante pruebas fisiológicas y bioquímicas de acuerdo con los esquemas de identificación indicados por MacFaddin. El perfil de susceptibilidad a los antibióticos de las colonias identificadas se realizó por el método de difusión en disco de Kirby Bauer. Se lograron identificar 2 especies de bacterias multirresistentes, entre ellas, cepas de Morganella morganii y Plesiomonas shigelloides resistentes a antibióticos de uso clínico como ceftazidima, aztreonam, ciprofloxacino, ácido nalidíxico y trimetroprim-sulfametoxazol. Los resultados muestran que las aguas del río Chibunga albergan bacterias con multirresistencia a los antimicrobianos, representando un riesgo de contaminación de los productos agrícolas cosechados en sus inmediaciones, así como para las personas que utilizan estas aguas.

RESUMEN Introducción: el lupus eritematoso sistémico (LES) se conoce como una enfermedad autoinmune multisistémica, cuya causa es desconocida, se caracteriza por la presencia de complejos inmunes y autoanticuerpos. En series internacionales se describe una presentación de procesos infecciosos en estos pacientes hasta en un 75 % de los casos, las infecciones ocurren tanto al inicio de la enfermedad como en etapas tardías, y son causa directa de muerte en 30 % a 60 % de los casos y motivo de hospitalización hasta de 30 %. Objetivos: determinar los procesos infecciosos más frecuentes en pacientes con lupus eritematoso sistémico internados en el Servicio de Clínica Médica del Hospital Nacional en el periodo 2022-2023. Metodología: diseño observacional, descriptivo de corte transversal de pacientes con diagnóstico de lupus eritematoso sistémico según criterios de European League Against Rheumatism (EULAR)-2019 que se encuentran internados en el servicio de Clínica Médica del Hospital Nacional (Itaugua-Paraguay) en el periodo de 2022-2023. Resultados: la edad media de los pacientes fue de 34 años, con predominio del sexo femenino en el 88.18 %. Las infecciones del aparato respiratorio fue la más frecuentemente diagnosticada en 50 (45.45 %) pacientes, seguido por la infección de vías urinarias 47 (42.38 %) pacientes, el condicionante de riesgo predominante fue el uso de corticoides en un total 105 (96.40 %) pacientes, se estableció en un total de 97 (88.18 %) pacientes con antibioticoterapia, dentro del aspecto demográfico de la zona urbana 56 (50.90 %) pacientes y rural 54(49.1 %). Conclusión: la infección del aparato respiratorio fue la infección más frecuente, el condicionante de riesgo predominante es el uso de corticoides y recibieron antibioticoterapia la cual en monoterapia fue la más utilizada

ABSTRACT Introduction: systemic lupus erythematosus (SLE) is known as a multisystem autoimmune disease, whose cause is unknown, and is characterized by the presence of immune complexes and autoantibodies. In international series, presentation of infectious processes is described in these patients in up to 75 % of cases. Infections occur both at the beginning of the disease and in late stages, and are a direct cause of death in 30 % to 60 % of patients. Cases and reason for hospitalization up to 30 %. Objectives: determine the most frequent infectious processes in patients with systemic lupus erythematosus admitted to the Medical Clinic Service of the Hospital Nacional in the period 2022-2023. Methodology: observational, descriptive cross-sectional design of patients with a diagnosis of systemic lupus erythematosus according to criteria of European League Against Rheumatism (EULAR)-2019 who are admitted to the Medical Clinic service of the Hospital Nacional (Itauguá-Paraguay) in the period of 2022-2023. Results: the average age of the patients was 34 years, with a predominance of the female sex in 88.18 %. Respiratory system infections were the most frequently diagnosed in 50 (45.45 %) patients, followed by urinary tract infection in 47 (42.38 %) patients. The predominant risk factor was the use of corticosteroids in a total of 105 (96.40 %) patients, it was established in a total of 97 (88.18 %) patients with antibiotic therapy, within the demographic aspect of the urban area 56 (50.90 %) patients and rural 54 (49.1 %). Conclusion: respiratory tract infection was the most frequent infection, the predominant risk factor is the use of corticosteroids and they received antibiotic therapy, which in monotherapy was the most used.

Abstract Poultry production is one of the most important agricultural sectors worldwide due to the high nutritional value of its products, such as meat and eggs, for human consumption. In this regard, veterinary antibiotics are used to treat or prevent disease-causing pathogens in order to ensure and maintain production. The objective of the study was to design and validate two questionnaires for assessing the risk of veterinary antibiotics used in egg-laying hens and their perceived impact in relation to food safety. Its logic and the validity of its content were determined by expert evaluation. Its construct validity was assessed by exploratory factor analysis, and its reliability, with Cronbach's Alpha coefficient. The survey was applied to 44 establishments or egg producers in the Espaillat province and to 385 consumers in the Santo Domingo province. A Cronbach's alpha coefficient of 0.799 was obtained for egg producers and veterinarians, and of 0.771, for consumers. The principal component analysis identified a KMO sample size adequacy measure of 0.558 for egg producers and veterinarians, and 0.797 for consumers. The questionnaire for egg producers and veterinarians consists of 8 factors and 22 items, and the questionnaire for consumers, of 3 factors and 8 items. The results confirm that the scale found is reliable and valid for the construct the risks associated with the potential consumption of food containing veterinary antibiotic residues.

Resumen La producción avícola es uno de los sectores agropecuarios de mayor importancia a nivel mundial por sus grandes aportes nutricionales en productos como la carne y el huevo para fines de la alimentación humana. En este sentido, para asegurar y mantener la producción se utilizan antibióticos veterinarios para tratar o prevenir agentes patógenos que causan enfermedades. El objetivo del estudio fue diseñar y validar dos cuestionarios para evaluar el riesgo de los antibióticos veterinarios de uso en gallinas ponedoras de huevos y su percepción de impacto con relación con la seguridad alimentaria. Se determinaron su lógica y validez de contenido mediante la evaluación por expertos. La validez de constructo se realizó mediante el análisis factorial exploratorio y la confiabilidad con el coeficiente de Alpha de Cronbach. Se aplicó a 44 establecimientos o productores de huevos en la provincia Espaillat y 385 consumidores de la provincia Santo Domingo. Se obtuvo un coeficiente de alfa de Cronbach de 0.799 para productores de huevos y veterinarias y 0.771 para los consumidores. El análisis de componente principales permitió identificar la medida de adecuación del tamaño de muestra KMO de 0.558 para los productores de huevos y veterinarias y 0.797 para los consumidores. El cuestionario está conformado por 8 factores y 22 ítems para los productores de huevos y veterinarias y 3 factores y 8 ítems para los consumidores. Los resultados confirman que la escala encontrada es confiable y válida para la construcción de los riesgos asociados al posible consumo de los alimentos con residuos de antibióticos veterinarios.

ABSTRACT Introduction. Inappropriate use of antimicrobials in the Intensive Care Unit (ICU) contributes to bacterial resistance. Objective. The purpose of the present study was to measure the use and type of antimicrobials in patients hospitalized in the Intensive Care Unit of the Loreto Regional Hospital (HRL). Methods. Descriptive study. We reviewed 120 medical records in ICU-HRL patients from January to June 2023. Results. Most patients used antimicrobials (74.2%), indicated for septic shock (42.7%), 1 to 3 antimicrobials (93.2%), with therapy of 1 to 3 days (45%). Of the total number of antimicrobials used, the greatest proportion were bactericides 89 (90.4%), from the cephalosporin family (33.1%) and carbapenemics (23.5%); the most commonly used drugs were ceftriaxone (26.5%) and meropenem (21.1%) by the intravenous route (90.4%). Conclusion. The highest proportion of hospitalized patients use antimicrobials of the cephalosporin and carbapenemics family.

RESUMEN Introducción. El uso inapropiado de antimicrobianos en las unidades de cuidados intensivos (UCI) contribuye a la resistencia bacteriana. Objetivo. El propósito del presente estudio fue medir el uso y tipo de antimicrobianos en los pacientes hospitalizados en la Unidad de Cuidados Intensivos del Hospital Regional de Loreto (HRL). Métodos. Estudio de tipo descriptivo. Revisamos 120 historias clínicas en pacientes de UCI-HRL desde enero a junio 2023. Resultados. La mayoría de los pacientes usaron antimicrobianos (74,2%), indicados por shock séptico (42,7%), de 1 a 3 antimicrobianos (93,2%), con terapia de 1 a 3 días (45%). Del total de antimicrobianos usados, la mayor proporción fueron bactericidas 89 (90,4%), de la familia de cefalosporinas (33,1%) y carbapenémicos (23,5%); los fármacos más empleados fueron ceftriaxona (26,5%) y meropenem (21,1%) por vía intravenosa (90,4%). Conclusión. La mayor proporción de pacientes hospitalizados usan antimicrobianos de la familia de cefalosporinas y carbapenémicos.

ABSTRACT Dengue is a viral infection which is transmitted by the Aedes aegypti mosquito and has four serotypes (DENV-1 to DENV-4). The disease triggers a variety of clinical manifestations, ranging from mild forms without warning signs to severe lifethreatening forms. We present the case of a 5-year-old boy, from the province of Callao, whose first symptoms were fever, headache and general malaise. On the third day, the child had mild abdominal pain and little vomiting; subsequently, abdominal distension, jaundice and choluria. He was admitted to the pediatric intensive care unit being alert and with moderate dehydration, jaundice, edema, distended and tender abdomen, shifting dullness and liver 2 cm below the right costal margin. Complementary tests revealed liver failure, hepatosplenomegaly and pleural effusion in the bases. Using a reactive IgM ELISA, severe dengue was diagnosed, as well as a superinfection due to probable spontaneous bacterial peritonitis. He started treatment with antibiotics, furosemide, fresh frozen plasma, cryoprecipitate and metamizole. As the child did not get better, the diuretic was optimized, and human albumin was administered. Thereafter, he got better showing decreased ascites, edema, jaundice and pleural effusion; improvement of the liver and coagulation profile; and being afebrile. He unexpectedly presented respiratory distress due to congestive heart failure caused by dilated cardiomyopathy diagnosed by echocardiography; thus, he was treated with diuretics. The patient was discharged afebrile, without edema and with resolution of liver failure and coagulation disorder.

RESUMEN El dengue es una infección viral transmitida a través del mosquito Aedes aegypti y presenta cuatro serotipos (DENV-1 a DENV-4). La enfermedad desencadena una variedad de manifestaciones clínicas, desde formas leves sin signos de alarma hasta formas graves, potencialmente mortales. Se presenta el caso de un niño de cinco años, procedente de la provincia del Callao, cuyos síntomas iniciales fueron fiebre, cefalea y malestar general. Al tercer día, el niño manifiestó dolor abdominal leve y vómitos escasos; posteriormente, distensión abdominal, ictericia y coluria. Fue hospitalizado en la unidad de cuidados intensivos pediátricos con deshidratación moderada, ictericia, edemas, abdomen distendido y doloroso, matidez desplazable, hígado a 2 cm debajo del reborde costal derecho y lúcido. Por exámenes complementarios, se evidenció falla hepática, hepatoesplenomegalia y derrame pleural en bases. Se diagnosticó dengue grave a través de una prueba de ELISA Ig M reactivo más sobreinfección por probable peritonitis bacteriana espontánea. Se inició el tratamiento con antibióticos, furosemida, plasma fresco congelado, crioprecipitado y metamizol. Al no observarse mejoría, se optimizó el diurético y se administró albúmina humana. Mostró mejoría con disminución de ascitis, edemas, ictericia y efusión pleural; también mejora del perfil hepático y de la coagulación, además de encontrarse afebril. Presentó inesperadamente dificultad respiratoria por insuficiencia cardiaca congestiva debido a miocardiopatía dilatada según ecocardiografía; se manejó con diuréticos. Fue dado de alta en estado afebril, sin edemas y con resolución de falla hepática y trastorno de coagulación.

ABSTRACT: This study addresses the need to understand the presence of functional microorganisms in tropical soils, providing valuable information for conservation and sustainable management. The main objective is to analyze the variability of functional microorganisms at different soil depths and their correlations with physicochemical parameters. To this end, soil samples were collected from 15 random points in a reserved forest and analyzed for physicochemical and microbiological properties at two depths (10 and 20 cm). The following functional microorganisms were quantified: viable aerobes, lactobacilli, actinomycetes, fungi, and nitrogen-fixing bacteria using specific protocols. The non-parametric Wilcoxon test was used to determine differences between depths, and the Pearson correlation coefficient (|ρ| ≥ 0.8) was used to analyze correlations between physicochemical parameters and microorganisms. The results suggest that the abundance of viable aerobic microorganisms, Lactobacillus, Actinomycetes, and Fungi decreases with soil depth, while nitrogen-fixing bacteria show no significant differences. The decrease in abundance is related to factors such as nutrient and oxygen availability and microenvironmental conditions. There is a strong positive correlation between the amount of viable aerobic microorganisms and Lactobacillus with phosphorus content, and a negative correlation between Actinomycetes and clay content. Actinomycetes prefer sandy soils, while Fungi thrive in soils rich in organic matter. The negative correlation between Fungi and Actinomycetes suggests competition for limited resources and possible chemical interactions, such as the production of antibiotic compounds by Actinomycetes. This study demonstrates that a detailed understanding of soil microbiota can offer new opportunities to improve agricultural practices, especially in terms of soil fertility management and environmental sustainability.

RESUMEN: Este estudio aborda la necesidad de entender la presencia microorganismos funcionales en suelos tropicales, proporcionando información valiosa para la conservación y manejo sostenible. Tiene como principal objetivo analizar la variabilidad de microorganismos funcionales en diferentes profundidades del suelo y las correlaciones con los parámetros fisicoquímicos. Para ello se recolectaron muestras de suelo de 15 puntos aleatorios en un bosque reservado y se analizaron propiedades fisicoquímicas y microbiológicas a dos profundidades (10 y 20 cm). Se cuantificaron los siguientes microorganismos funcionales: aerobios viables, lactobacilos, actinomicetos, hongos y bacterias fijadoras de nitrógeno utilizando protocolos específicos. Se empleó la prueba no paramétrica de Wilcoxon para determinar diferencias entre profundidades y el coeficiente de correlación de Pearson (|ρ| ≥ 0,8) para analizar correlaciones entre parámetros fisicoquímicos y microorganismos. Los resultados sugieren que la abundancia de microorganismos aerobios viables, Lactobacillus, Actinomicetos y Fungi disminuye con la profundidad del suelo, mientras que las bacterias fijadoras de nitrógeno no muestran diferencias significativas. La disminución de la abundancia se relaciona con factores como la disponibilidad de nutrientes y oxígeno y las condiciones microambientales. Existe una correlación positiva fuerte entre la cantidad de microorganismos aerobios viables y Lactobacillus con el contenido de fósforo, y una correlación negativa entre Actinomicetos y el contenido de Arcilla. Los Actinomicetos prefieren suelos arenosos, mientras que los Fungi prosperan en suelos ricos en materia orgánica. La correlación negativa entre Fungi y Actinomicetos sugiere competencia por recursos limitados y posibles interacciones químicas, como la producción de compuestos antibióticos por parte de los Actinomicetos. Este estudio demuestra que la comprensión detallada de la microbiota del suelo puede ofrecer nuevas oportunidades para mejorar las prácticas agrícolas, especialmente en términos de gestión de la fertilidad del suelo y sostenibilidad ambiental.

ABSTRACT: More and more countries are incorporating new safety, health and food safety regulations, responding to the growing concern about the use of antibiotic growth promoters (APC) in animal production, the scientific community has been forced to seek new alternatives that reduce or replace the use of (APC), with the ability to safeguard public health. Within these alternatives, phytobiotics are considered a very promising option, since they are natural substances made up of complex bioactive compounds. This document reviews the different applications and functionality of phytobiotics in animal production. Firstly, the main effects and mechanism of action are shown, highlighting and explaining their functionality as antioxidants, anti-inflammatory, antimicrobial, growth promoters, improvers of the organoleptic characteristics of foods and reduces the generation of gases that cause the greenhouse effect. Second, notable applications of phytobiotics to different species of zootechnical interest and their effect are reviewed. Finally, their bioavailability and the current and future challenges of phytobiotics are investigated.

RESUMEN: Cada vez más países van incorporando nuevas normativas de inocuidad, seguridad sanitaria y alimentaria, respondiendo a la progresiva preocupación por el uso de antibióticos promotores de crecimiento (APC) en producción animal; la comunidad científica se ha visto obligado a buscar nuevas alternativas que reduzcan o sustituyan el uso de APC, con la capacidad de salvaguardar la salud pública. Dentro de esas alternativas, los fitobióticos son considerados una opción muy prometedora, ya que son sustancias naturales constituidas por complejos compuestos bioactivos. Este documento revisa las distintas aplicaciones y la funcionalidad de los fitobióticos en la producción animal. En primer lugar, se manifiesta los principales efectos y mecanismo de acción, destacando y explicando su funcionalidad como antioxidantes, antiinflamatorios, antimicrobianos, promotores de crecimiento, mejoradores de las características organolépticas de los alimentos y reductores de la generación de gases causantes del efecto invernadero, Segundo, se revisan aplicaciones destacadas de los fitobióticos a las distintas especies de interés zootécnico y su efecto que estos tienen. Finalmente, se indaga su biodisponibilidad y los desafíos actuales y futuros de los fitobióticos.

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 impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O presente estudo foi realizado para isolar e caracterizar bactérias de amostras de água e solo retiradas do Canal Lahore, em Lahore, em diferentes locais, ou seja, Mall Road, Mohlanwal e Khera. As cepas bacterianas isoladas foram identificadas com base em testes morfológicos e bioquímicos. A identificação foi confirmada por cultura de bactérias em testes de meios seletivos. O teste de resistência aos antibióticos também foi realizado para observar a resistência das bactérias a diferentes antibióticos. Foi realizado o teste de ágar sangue para identificar diferentes bactérias patogênicas. O resultado revelou que amostras de água e solo do Canal Lahore, Lahore, de diferentes localidades estavam contaminadas com Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. e Staphylococcus spp. Por causa da presença desses patógenos, essa água não é adequada para qualquer uso doméstico e de irrigação. O estudo revelou que a água do Canal Lahore é prejudicial à saúde humana, pois está contaminada com bactérias que podem causar doenças graves, por exemplo: Escherichia coli pode ocasionar gastroenterite; Bacillus spp. pode causar náuseas e vômitos; Enterococcus sp. pode infectar o trato urinário; Salmonella sp. é responsável pela bacteremia; Staphylococcus spp. pode causar febre leve; e Vibrio sp. pode ser a razão da cólera. Assim, torna-se imprópria para uso humano, como natação, banho, lavagem etc., até que algumas medidas corretivas sejam empregadas para erradicar microrganismos patogênicos por WASA e LWMS de acordo com os padrões da OMS. Da mesma forma, é bastante prejudicial, quando usada para irrigação sem tratamento adequado.

Abstract The present study was conducted to isolate and characterize bacteria from water and soil sample taken from the Lahore Canal at different sites i.e. Mall Road, Mohlanwal and Khera site. Isolated bacterial strains were identified on the basis of morphological and biochemical tests. Identification was confirmed by culturing bacteria on selective media. Antibiotic resistance test was also performed to observe the resistance of bacteria against different antibiotics. Blood agar test was performed for identification of different pathogenic bacteria. The result revealed that water and soil samples of Lahore Canal Lahore from different sites were contaminated with Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. and Staphylococcus spp. Due to presence of these pathogens, this water is not suitable for any domestic and irrigation use. Study also revealed that water of the Lahore Canal is harmful for human health as it is contaminated with bacteria that can cause severe disease e.g., Escherichia coli can cause gastroenteritis, Bacillus spp. can cause nausea and vomiting, Enterococcus may infect urinary tract, Salmonella sp. is responsible for Bacteremia, Staphylococcus spp. can cause mild fever and Vibrio sp. can be the reason of cholera. Thus it is rendered unfit for any kind of human use even other than drinking like swimming, bathing, washing etc., until and unless some remedial measures are employed to eradicate pathogenic microorganisms by WASA and LWMS according to standards of WHO. Similarly, it is quite harmful, when and where ever it is used for irrigation without proper treatment.

Resumo A produção de plantas transgênicas com características agronômicas e hortícolas desejadas vem ganhando grande importância por atender às demandas da crescente população. A transformação genética também é um passo fundamental para estudar os fundamentos das ciências das plantas. Foram desenvolvidos e utilizados diferentes protocolos de transformação, que são confiáveis e eficientes. Esses protocolos usaram genes de resistência a antibióticos ou herbicidas incorporados ao gene de interesse para identificar plantas transformadas e não transformadas. Esses genes marcadores podem representar uma ameaça ao ser humano e ao meio ambiente. O uso de marcadores visuais permite a observação direta e fácil de plantas transformadas com mais precisão. No estudo atual, foi projetado um cassete de genes com o gene da hidroxilase de produção de pigmentos (PPH) sob promotor específico de fibra (GhSCFP) e terminador NOS a jusante. Após verificar a eficiência estrutural e funcional do gene otimizado por códons usando ferramentas de bioinformática, o cassete foi enviado para síntese química a partir de fonte comercial. O cassete do gene do pigmento (PPH_CEMB), clonado em pCAMBIA-1301, foi transformado em Agrobacterium por eletroporação. O método de imersão floral mediado por Agrobacterium foi usado para transformar a inflorescência de Camelina sativa. Após a formação de sementes, foi observado um total de 600 sementes com mudança de cor, das quais 19 desenvolveram uma coloração marrom-avermelhada, enquanto as 581 restantes permaneceram amarelas. A eficiência de transformação calculada com base na mudança de cor foi de 1%. A análise de PCR das folhas obtidas após a semeadura de sementes avermelhadas confirmou a transformação do gene produtor de pigmentos, enquanto não foi observada amplificação por PCR em folhas de plantas de sementes do tipo selvagem. A partir dos resultados, é evidente que a transformação da inflorescência de C. sativa mediada por Agrobacterium é uma técnica muito eficiente e favorável ao ambiente não só para a detecção de plantas transformadas, mas também para estudar processos celulares básicos.

Abstract Production of transgenic plants with desired agronomic and horticultural traits has gained great importance to fulfill demands of the growing population. Genetic transformation is also a fundamental step to study basics of plant sciences. Different transformation protocols have been developed and used which are reliable and efficient. These protocols used antibiotic or herbicide resistance genes incorporated along with gene of interest to identify transformed plants from non-transformed ones. These marker genes may pose a threat to human and environment. Use of visual markers enables direct and easier observation of transformed plants with more precision. In current study a gene cassette with ‘pigment production hydroxylase (PPH) gene under fiber specific promoter (GhSCFP) and downstream Nos-terminator was designed. After checking the structural and functional efficiency of codon optimized gene using bioinformatics tools, the cassette was sent for chemical synthesis from commercial source. The pigment gene cassette (PPH_CEMB), cloned in pCAMBIA-1301, was transformed into Agrobacterium through electroporation. Agrobacterium-mediated floral dip method was used to transform Camelina sativa inflorescence. After seed setting a total of 600 seed were observed for change in color and out of these, 19 seeds developed a reddish-brown coloration, while the remaining 581 seeds remained yellow. The transformation efficiency calculated on basis of color change was 1.0%. PCR analysis of leaves obtained after sowing reddish seeds confirmed the transformation of pigment production gene, while no PCR amplification was observed in leaves of plants from wild type seeds. From the results it is evident that Agrobacterium-mediated transformation of C. sativa inflorescence is very efficient and environment friendly technique not only for detection of transformed plants but also to study basic cellular processes.

Resumo A Escherichia coli patogênica aviária (APEC) induz a colibacilose, uma doença aguda e sistêmica, resultando em perdas econômicas substanciais no setor avícola. Este estudo teve como objetivo investigar o padrão de resistência a antibióticos associado à frequente distribuição de genes de virulência em APEC O78:K80 que podem causar alterações patológicas em galinhas. O perfil do antibiograma mostrou alta resistência à eritromicina, cloranfenicol, tetraciclina, ampicilina e cotrimoxazol; resistência intermediária à ciprofloxacina, levofloxacina, enrofloxacina, norfloxacina, nitrofurantoína e cloridrato de doxiciclina; e sensível à amicacina, estreptomicina, gentamicina e colistina . A distribuição de genes de virulência identificou oito genes (irp-2, iutA, ompT, iss, iucD, astA, hlyF e iroN) por meio de uma reação em cadeia da polimerase convencional. A APEC O78:K80 causou concentrações significativamente altas de enzimas hepáticas, níveis séricos de interleucina-6 e fator de necrose tumoral alfa em aves experimentais. Além disso, aves infectadas apresentaram hipoproteinemia, hipoalbuminemia e hiperglobulinemia. O exame de necropsia revelou peri-hepatite e pericardite fibrinosa, pulmões congestos, hemorragias equimóticas do intestino e granulomatose necrosante do baço. O exame histopatológico mostrou degeneração hepatocelular, necrose miocárdica, nefrite intersticial, hemorragias intestinais e linfopenia no baço. Este estudo é a primeira evidência para avaliar o perfil de resistência a antibióticos associado a genes de virulência e potencial clínico-patológico de APEC O78:K80 em galinhas no Paquistão, o que pode ser uma abordagem útil e rápida para prevenir e controlar a doença por meio do desenvolvimento de estratégias de controle.

Abstract Avian pathogenic Escherichia coli (APEC) induces colibacillosis, an acute and systemic disease, resulting in substantial economic losses in the poultry sector. This study aimed to investigate the antibiotic resistance pattern associated with frequent virulence gene distribution in APEC O78:K80 that may cause pathological alterations in chickens. The antibiogram profile showed high resistance to erythromycin, chloramphenicol, tetracycline, ampicillin, and co-trimoxazole, followed by intermediate resistance to ciprofloxacin, levofloxacin, enrofloxacin, norfloxacin, nitrofurantoin, and doxycycline hydrochloride, and sensitive to amikacin, streptomycin, gentamicin, and colistin. Virulence gene distribution identifies eight (irp-2, iutA, ompT, iss, iucD, astA, hlyF, iroN) genes through a conventional polymerase chain reaction. APEC O78:K80 caused significantly high liver enzyme concentrations, serum interleukin-6 and tumor necrosis factor-alpha levels in experimental birds. Also, infected birds have hypoproteinemia, hypoalbuminemia, and hyperglobulinemia. Necropsy examination revealed fibrinous perihepatitis and pericarditis, congested lungs, intestinal ecchymotic hemorrhages and necrotizing granulomatosis of the spleen. Histopathological examination depicted hepatocellular degeneration, myocardial necrosis, interstitial nephritis, intestinal hemorrhages and lymphopenia in the spleen. This study is the first evidence to assess the antibiotic resistance profile linked with virulence genes and clinicopathological potential of APEC O78:K80 in chickens in Pakistan, which could be a useful and rapid approach to prevent and control the disease by developing the control strategies.

Resumo Devido à extensa aplicação de antibióticos como promotores de crescimento na alimentação animal, a resistência aos antimicrobianos aumentou. Para superar esse desafio, os microbiologistas do rúmen buscam novos probióticos para melhorar a produtividade do gado. O presente estudo teve como objetivo isolar e avaliar bactérias lácticas específicas de raças (BAL) como potenciais probióticos animais. 12 cepas foram isoladas de diferentes amostras, incluindo leite, rúmen e fezes de búfalos Nilli Ravi. Esses isolados foram avaliados quanto ao seu potencial antimicrobiano contra patógenos animais comuns (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). Todos os isolados foram identificados por meio do sequenciamento do gene 16S rRNA e as análises filogenéticas inferiram que essas cepas apresentaram estreita relação com as espécies de vários gêneros; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis, Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis e Lactococcus lactis. O perfil probiótico dos isolados foi obtido usando critérios padrão, particularmente com propriedades probióticas animais.

Abstract Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties

Resumo Na biotecnologia moderna, o uso de enzimas para obter compostos novos ou modificados com propriedades antibacterianas, antifúngicas e anticancerígenas é crucial. Lactases de cogumelos são biocatalisadores promissores para síntese e modificação de diferentes compostos, por serem enzimas baratas e disponíveis para a preparação de componentes de reação, e vem recebendo a devida atenção recentemente. A purificação da lacase foi realizada a partir do basidiomiceto Lentinus strigosus em vários estágios: Etapa 1 - na cromatografia de troca iônica em TEAE Servacell 23 (400 ml), foram observados dois picos de atividade da lacase distintamente separados, com eluição do transportador a 0,21 e 0,27 M de NaCl. Para reduzir a perda de enzimas, todas as frações com atividade de lacase foram coletadas, concentradas e dessalinizadas em uma célula de ultrafiltração (Amicon, Estados Unidos) com membrana UM-10; Etapa 2 - a preparação resultante com atividade de lacase foi aplicada a uma coluna Q-Sepharose (60 ml). Durante a eluição, foram obtidos dois picos bem separados com atividade de lacase: lacase I (NaCl 0,12 M) e lacase II (NaCl 0,2 M); Etapa 3 - no decurso da purificação adicional de ambas as enzimas, realizada no Recurso Q de transportador de troca aniônica (6 ml), um gradiente quebrado foi usado: 0-10%, 10-20% e 20-100% com NaCl 1M; Etapa 4 - tanto a lacase I como a lacase II, obtidas após o Recurso Q, foram dessalinizadas e concentradas para 1 ml cada e aplicadas a uma coluna de filtração em gel Superdex 75. Como resultado, duas lacases foram obtidas de forma homogênea.

Abstract In advanced biotechnology, the utilization of enzymes to achieve new or modified compounds with antibacterial, fungicidal, and anti-cancer specifications is crucial. Mushroom lactases are a hopeful biocatalyst for the synthesis and modification of different compounds. They are an accessible and inexpensive enzyme for the preparation of reaction objects and have recently received attention. Laccase purification was performed from basidiomycete Lentinus strigosus (LS) in several stages: Stage 1. On ion-exchange chromatography on TEAE Servacell 23 (400 ml), two distinctly separated laccase activity peaks were observed, eluted from the carrier at 0.21 and 0.27 M NaCl. In order to reduce the loss of enzymes, all fractions with laccase activity were collected, concentrated, and desalted using an ultrafiltration cell (Amicon, United States) with a UM-10 membrane. Stage 2. The resulting preparation with laccase activity was applied to a Q-Sepharose column (60 ml). Two well-separated peaks with laccase activity were obtained during the elution: laccase I (0.12 M NaCl) and laccase II (0.2 M NaCl). Stage 3. In the course of further purification of both enzymes, carried out on anion-exchange carrier Resource Q (6 ml), a broken gradient was used: 0 - 10%, 10 - 20%, and 20 - 100% with 1M NaCl. Stage 4. Both laccase I and laccase II, obtained after Resource Q, were desalted, concentrated to 1 ml each, and applied to a Superdex 75 gel filtration column. As a result, two laccases were obtained in a homogeneous form.