A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.

Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples

In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43), cattle diarrheic faeces (48) and poultry faecal swabs (52) only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30), milk (35), cheese (30), only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each) as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.

Unraveling the signaling and signal transduction mechanisms controlling arbuscular mycorrhiza development

Arbuscular mycorrhiza (AM) are the most widespread symbiotic associations between plant roots and soil fungi. AM can contribute to increasing the survival and fitness of plants to limiting environments mostly due to their ability in improving nutrient uptake from the soil solution. Despite their ecological significance, the mechanisms controlling AM development and functioning are largely unknown. The obligate mutualistic nature of the arbuscular mycorrhizal fungi (AMF) has hampered the advances on the understanding and application of the symbiosis. Significant alterations in the genetic programs of both symbionts are required for the successful establishment of an AM, and complex signaling and signal transduction mechanisms are likely involved. The analyses of legume mutants affected in the development of nitrogen fixing nodules and AM suggest that part of the signal transduction pathways involved in the regulation of both symbioses are conserved. Even though the use of genomics of model plants has helped to advance our understanding of the regulatory mechanisms in AM, identifying the signal molecules involved in plant-AMF communication and determining their transduction pathways is still essential for its biotechnological application in agriculture.

As micorrizas arbusculares (MAs) são as associações simbióticas entre raízes de plantas e fungos mais comuns na natureza. Elas podem contribuir para o aumento da sobrevivência e adaptação das plantas a ambientes limitantes, principalmente devido a sua maior capacidade em absorver nutrientes da solução do solo. Apesar de sua importância ecológica, os mecanismos que controlam o desenvolvimento e fisiologia das MAs são pouco conhecidos. A natureza mutualística obrigatória dos fungos micorrízicos arbusculares (FMAs) tem limitado os avanços na compreensão e aplicação da simbiose. Alterações significativas nos programas genéticos de ambos simbiontes são necessárias para o estabelecimento de MAs, e mecanismos complexos de sinalização e transdução de sinais estão provavelmente envolvidos. A análise de mutantes de leguminosas defectivos no desenvolvimento de nódulos fixadores de nitrogênio e MAs sugere que uma parte das vias de transdução de sinais envolvidas na regulação dessas simbioses é conservada. Muito embora o uso da genômica de plantas modelos tenha contribuído para o avanço na compreensão dos mecanismos que regulam MAs, a identificação de moléculas sinais envolvidas na comunicação planta-FMA, e a determinação de suas vias de transdução, é essencial para aplicação da simbiose na agricultura.

Unraveling the signaling and signal transduction mechanisms controlling arbuscular mycorrhiza development

Arbuscular mycorrhiza (AM) are the most widespread symbiotic associations between plant roots and soil fungi. AM can contribute to increasing the survival and fitness of plants to limiting environments mostly due to their ability in improving nutrient uptake from the soil solution. Despite their ecological significance, the mechanisms controlling AM development and functioning are largely unknown. The obligate mutualistic nature of the arbuscular mycorrhizal fungi (AMF) has hampered the advances on the understanding and application of the symbiosis. Significant alterations in the genetic programs of both symbionts are required for the successful establishment of an AM, and complex signaling and signal transduction mechanisms are likely involved. The analyses of legume mutants affected in the development of nitrogen fixing nodules and AM suggest that part of the signal transduction pathways involved in the regulation of both symbioses are conserved. Even though the use of genomics of model plants has helped to advance our understanding of the regulatory mechanisms in AM, identifying the signal molecules involved in plant-AMF communication and determining their transduction pathways is still essential for its biotechnological application in agriculture.

As micorrizas arbusculares (MAs) são as associações simbióticas entre raízes de plantas e fungos mais comuns na natureza. Elas podem contribuir para o aumento da sobrevivência e adaptação das plantas a ambientes limitantes, principalmente devido a sua maior capacidade em absorver nutrientes da solução do solo. Apesar de sua importância ecológica, os mecanismos que controlam o desenvolvimento e fisiologia das MAs são pouco conhecidos. A natureza mutualística obrigatória dos fungos micorrízicos arbusculares (FMAs) tem limitado os avanços na compreensão e aplicação da simbiose. Alterações significativas nos programas genéticos de ambos simbiontes são necessárias para o estabelecimento de MAs, e mecanismos complexos de sinalização e transdução de sinais estão provavelmente envolvidos. A análise de mutantes de leguminosas defectivos no desenvolvimento de nódulos fixadores de nitrogênio e MAs sugere que uma parte das vias de transdução de sinais envolvidas na regulação dessas simbioses é conservada. Muito embora o uso da genômica de plantas modelos tenha contribuído para o avanço na compreensão dos mecanismos que regulam MAs, a identificação de moléculas sinais envolvidas na comunicação planta-FMA, e a determinação de suas vias de transdução, é essencial para aplicação da simbiose na agricultura.