Biological indexing is the method generally recognized for the certification of propagative grapevines in many countries, and it is mandatory in the European Union. It consists of the evaluation of the plant material after grafting on indicators that are inspected for symptom development. This is a lengthy process that requires well-trained workers, testing field, etc. Alternative diagnostic methods such as serology and RT-qPCR have been discarded for certification because of their intrinsic drawbacks. In turn, high-throughput sequencing (HTS) of plant RNA has been proposed as a plausible alternative to bioassay, but before it is accepted, different aspects of this process must be evaluated. We have compared the HTS of small RNAs with bioassays and other diagnostic methods from a set of 40 grapevine plants submitted for certification. The results allowed the authors the identification of numerous grapevine viruses in the samples, as well as different variants. Besides, relationships between symptom expression and viromes were investigated, in particular leafroll-associated viruses. We compared HTS results using analytical and bioinformatics approaches in order to define minimum acceptable quality standards for certification schemes, resulting in a pipeline proposal. Finally, the comparison between HTS and bioassay resulted favorable for the former in terms of reliability, cost, and timing.
Hepatitis E virus (HEV) is an important enteric agent that can circulate in swine; it is excreted in manure, and of zoonotic interest. The present study investigated, by RT-qPCR, the circulation of HEV in swine manure from different types of pig farms (maternity, nursery, and grow-finish farms) in Santa Catarina State, the major pig production area of Brazil, and also evaluated the HEV removal efficiency of psychrophilic anaerobic biodigesters (PABs). While HEV was consistently detected in manure from grow-finish pig farms (>4 log HEV genome copies (GC) L(−1)), the virus was not detected in manure from maternity and nursery farms. These findings suggest a potential high biosafety status during primary-swine production, with a subsequent contamination in grow-finish production. The anaerobic biodigestion process reduced more than 2 log(10) HEV GC in the processed swine manure. However, the virus concentration in final effluent remained high, with an average value of 3.85 log(10) HEV GC L(−1). Consequently, our results demonstrate that PABs can be a robust tool for effective inactivation of HEV, while reinforcing the need for sanitary surveillance and legislation of swine manure-derived biofertilizers, to avoid the spread of zoonotic enteric pathogens such as HEV.
Upaya strategis Pemerintah Kabupaten Sampang dalam meningkatkan kualitas Sumber Daya Manusia (SDM) yaitu dengan proses pemberdayaan masyarakat, dalam hal ini Pemerintah Kabupaten Sampang mengeluarkan kebijakan Program Kegiatan Peningkatan Sarana dan Prasarana Pendukung Lingkungan Sehat Perumahan Kecamatan Sampang, Kabupaten Sampang Tahun 2018. Program tersebut dimaksudkan untuk memberikan keleluasaan bagi masyarakat di tingkat RT untuk mengajukan kegiatan berbasis pemberdayaan masyarakat. Tujuan penelitian ini yaitu untuk menganalisis dan mendeskripsikan Implementasi Program Kegiatan Peningkatan Sarana dan Prasarana Pendukung Lingkungan Sehat Perumahan pada RT 02 RW 04 dan RT 02 RW 02 Kelurahan Dalpenang, ditinjau berdasarkan teori implementasi Bottom Up yaitu identifikasi jaringan aktor yang terlibat, jenis kebijakan publik yang mendorong masyarakat mengerjakan sendiri implementasi kebijakannya atau masih melibatkan pejabat pemerintah di level bawah, kebijakan yang dibuat sesuai dengan harapan keinginan publik yang menjadi target, prakarsa masyarakat secara langsung atau melalui lembaga swadaya masyarakat. Penelitian ini menggunakan jenis penelitian deskriptif dengan pendekatan kualitatif. Teknik pengumpulan data melalui observasi, wawancara, dan dokumentasi. Hasil penelitian menunjukkan, implementasi program tersebut telah dilaksanakan dengan jaringan aktor yang terlibat yaitu Tim Fasilitasi Kegiatan, Lurah Dalpenang, Ketua RT, dan tokoh masyarakat setempat. Jenis kebijakan publik yang mendorong masyarakat mengerjakan sendiri implementasi kebijakannya ditunjukkan dengan pekerja yang dapat diambil dari lingkungan RT setempat sedangkan birokrat level bawah yaitu Lurah Dalpenang hanya sebagai fungsi koordinasi. Adanya Musrembang di RT 02 RW 04 terkait program pemasangan penerangan jalan umum dan RT 02 RW 02 terkait program rehab jalan rabat beton menunjukkan program tersebut disesuaikan dengan kebutuhan riil masyarakat setempat. Saran peneliti dalam penelitian ini yaitu diharapkan kedepan Pemerintah Kabupaten Sampang dapat mengalokasikan anggaran kegiatan yang lebih besar, memperkuat tugas, tanggung jawab serta koordinasi pelaksana kegiatan dengan Tim Fasilitasi kegiatan, sehingga kebijakan program ini dapat dilaksanakan secara masif di tahun-tahun selanjutnya. Kata Kunci: Implementasi, Program Peningkatan Sarana Prasarana Strategic effort of Sampang Regency Givernment in improving the quality of Human Resources (HR) is through the process of community empowerment, in this case Sampang district givernment issued a program of Activities to Increase Facilities and Infrastructure Supporting the Healthy Environment for Housing in Sampang District, Sampang Regency in 2018. The program was intended to provide flexibility for the community at the RT level to propose community empowerment based activities. The purpose of this study is to analyze and describe the Implementation of the Program for the Improvement of Facilities and Infrastructure of Supporting Healthy Environments for Housing in RT 02 RW 04 and RT 02 RW 02 Dalpenang, reviewed based on Bottom Up implementation theory, namely identification of the network of actors involved, types of public policies that encourage people do their own policy implementation by themselves or still involve government officials at lower levels, policies made in accordance with the expectations of target public, community initiatives directly or through nongovernmental organizations. This research uses descriptive research with a qualitative approach. Data collection techniques through observation, interviews, and documentation. The results shows that the implementation of the program had been carried out with a network of actors involved, namely theActivity Facilitation Team, Dalpenang Village Chief, RT Chairperson, and local community leaders. The type of public policy that encourages the community to carry out its own policy implementation is shown by workers who can be taken from the local RT environment while the lower level bureaucrats namely the Dalpenang urban village leaders are only a function of coordination. The existence of Musrembang in RT 02 RW 04 related to the public street lighting installation program and RT 02 RW 02 related to the concrete rebate road rehabilitation programs show that the program is adjusted to the real needs of the local community. Researcher's suggestion in this research is that in the future the Sampang Regency Government can allocate a larger budget for activities, strengthen its tasks, responsibilities and coordinate the implementation of activities with the Facilitation Team of activities, so that this program's policies can be carried out massively in the following years. Keywords: Implementation, Infrastructure Facility Improvement Program.
Milking production is currently facing new challenges such as the end of the limit of annual milk's production in the European Union and the realization of the impact on the environment of cow's breeding. On the other hand, the industry has to produced more to answer the increase of dairy products. Thus the research on dairy cow and especially the bovine mammary gland is important. The mammary gland is a dynamic organ composed by different populations such as mammary stem cells, progenitor and differentiated cells, myoepithelial cells, adipocytes, fibroblasts and endothelial cells. All these population of cells contributed to milking production. Some surface cells markers such as CD49f, CD24, CD10 and EpCAM and the KRT49 gene are used to phenotype these cells. We used these makers to characterize the mammary stem progenitor and differentiated cells and myoepithelial cells during lactation and between two stages of development (heifer / lactating cow). We discovered that most of the populations stained were constant during our experiments. One population with regenerative properties decreased during lactation and the other variations concerned epithelial cell type at the beginning of lactation and were linked to the quantity of milk production. This stability among the cells could be due to the fixation of cell populations during the development of the mammary gland. To test this hypothesis, a work on a 7 month-old heifer must be done. On the other hand, the stem cells' reservoir could be situated in the adipocytes population. An extensive study of this population of cell should improve our understanding of the evolution of the mammary gland. ; La recherche sur la fonction de lactation et notamment sur le fonctionnement de la glande mammaire bovine est un outil indispensable pour inscrire la filière laitière dans la durabilité tout en lui permettant de faire face aux défis auxquels elle est confrontée comme l'arrêt des quotas laitiers ou la prise de conscience de l'impact environnemental de l'élevage bovin. La glande mammaire est en effet un organe dynamique composé de plusieurs populations de cellules qui contribuent à la production de lait : les cellules épithéliales souches, progénitrices et sécrétrices, les myoépithéliales, les fibroblastes, les adipocytes et les cellules endothéliales. Différents marqueurs tels que CD49f, CD24, CD10 et EpCAM ou le gène KRT19 permettent de phénotyper ces cellules par la technique de la cytométrie en flux et la RT-qPCR. Au cours de ce stage, nous avons utilisé ces différents marqueurs pour phénotyper les cellules épithéliales souches, progénitrices, sécrétrices et myoépithéliales au cours de la lactation d'une part et entre deux stades physiologiques contrastés (génisse / vache en lactation) d'autre part. Les résultats nous montrent que la majorité de ces populations restent stables au cours des échelles de temps étudiées. Pour les populations présentant des variations, elles concernent d'une part une population impliquée dans le renouvellement mammaire qui diminue au cours de la lactation et d'autre part des populations de type épithélial qui permettent d'expliquer les différences de production en début de lactation. Ces résultats nous ont amené sur de nouvelles hypothèses de travail parmi lesquelles les populations de cellules de la glande mammaire seraient fixées au cours du développement de cet organe ce qui nous amènerait à travailler sur la génisse de 7 mois et d'autre part que les adipocytes pourraient jouer un rôle de réservoir des cellules souches.
Los debates metodológicos en la ciencia política estadounidense se han profundizado a lo largo de las últimas décadas, marcadas por el Movimiento Perestroika de contestación de preferencias teóricas y metodológicas en la disciplina en Estados Unidos y, recientemente, por el esfuerzo de aprobación de una política de transparencia y acceso a datos, la Data Access and Research Transparency (DA-RT). La iniciativa ha dado lugar a que investigadores cualitativos y cuantitativos adopten distintas posturas con respecto a los significados de ciencia, transparencia y la propia profesión del politólogo. En este contexto, nuestro objetivo es ofrecer un panorama de los discursos sobre la DA-RT ante el debate metodológico subyacente a su implementación y sus potenciales desdoblamientos en la ciencia política en América Latina.
Diabetes mellitus is a form of metabolic disorder where patients are incapable to organize glucose metabolism. The most common types are Type I and Type II, constituting about 10% and 90% of cases, respectively. The cause of type I diabetes, which usually spreads in children and adolescents, is the disability of the endocrine system to produce insulin. On the other hand, The most common type of diabetes, type II diabetes, is often presented in adults. It is usually presented as a collection of insulin deficiency and insulin resistance. This work was done to estimate the count of microbiota in diabetics to find an appraoch for detection and follow-up treatment. The count of two types of bacteria Lactobacillus and Bifidobacterium was determined using qPCR based on the standard curve that was created from the serial decimal dilution of samples containing an unknown number of bacteria taken from probiotic capsules. The main results of this study show that the Lactobacillus count was affected by diabetes types, where a decrease was observed in the mean value in the case of diabetes type I group (32978.13) compared with the control group (610680.26). The mean value in diabetes type II was close to that of the control group (682199.27). While, the count of the Bifidobacterium showed a significant reduction in the mean value in both type I and type II diabetes groups (7521.70, 51880.82, respectively), compared with the control group (63405999.00).
Abstract Rapid and specific detection of extended-spectrum β-lactamase-producing (ESBL) bacteria is crucial both for timely antibiotic therapy when treating infected patients as well as for appropriate infection control measures aimed at curbing the spread of ESBL-producing isolates. Whereas a variety of phenotypic methods are currently available for ESBL detection, they remain time consuming and sometimes difficult to interpret while being also affected by a lack of sensitivity and specificity. Considering the longer turnaround time (TAT) of susceptibility testing and culture results, DNA-based ESBL identification would be a valuable surrogate for phenotypic-based methods. Putative ESBL-positive Enterobacteriaceae isolates (n = 330) from clinical specimen were prospectively collected in Bulgaria, Romania and Democratic Republic of Congo and tested in this study. All isolates were assessed for ESBL-production by the E-test method and those giving undetermined ESBL status were re-tested using the combination disk test. A genotypic assay successively combining qPCR detection of bla CTX-M, bla TEM and bla SHV genes with a multiplex pyrosequencing of bla TEM and bla SHV genes was developed in order to detect the most common ESBL-associated TEM and SHV single nucleotides polymorphisms, irrespective of their plasmid and/or chromosomal location. This assay was applied on all Enterobacteriaceae isolates (n = 330). Phenotypic and genotypic results matched in 324/330 (98.2%). Accordingly, real-time PCR combined with multiplex pyrosequencing appears to be a reliable and easy-to-perform assay with high-throughput identification and fast TAT (~5 h).
Rapid and specific detection of extended-spectrum β-lactamase-producing (ESBL) bacteria is crucial both for timely antibiotic therapy when treating infected patients as well as for appropriate infection control measures aimed at curbing the spread of ESBL-producing isolates. Whereas a variety of phenotypic methods are currently available for ESBL detection, they remain time consuming and sometimes difficult to interpret while being also affected by a lack of sensitivity and specificity. Considering the longer turnaround time (TAT) of susceptibility testing and culture results, DNA-based ESBL identification would be a valuable surrogate for phenotypic-based methods. Putative ESBL-positive Enterobacteriaceae isolates (n = 330) from clinical specimen were prospectively collected in Bulgaria, Romania and Democratic Republic of Congo and tested in this study. All isolates were assessed for ESBL-production by the E-test method and those giving undetermined ESBL status were re-tested using the combination disk test. A genotypic assay successively combining qPCR detection of blaCTX-M, blaTEM and blaSHV genes with a multiplex pyrosequencing of blaTEM and blaSHV genes was developed in order to detect the most common ESBL-associated TEM and SHV single nucleotides polymorphisms, irrespective of their plasmid and/or chromosomal location. This assay was applied on all Enterobacteriaceae isolates (n = 330). Phenotypic and genotypic results matched in 324/330 (98.2%). Accordingly, real-time PCR combined with multiplex pyrosequencing appears to be a reliable and easy-to-perform assay with high-throughput identification and fast TAT (~5 h).
Rapid and specific detection of extended-spectrum β-lactamase-producing (ESBL) bacteria is crucial both for timely antibiotic therapy when treating infected patients as well as for appropriate infection control measures aimed at curbing the spread of ESBL-producing isolates. Whereas a variety of phenotypic methods are currently available for ESBL detection, they remain time consuming and sometimes difficult to interpret while being also affected by a lack of sensitivity and specificity. Considering the longer turnaround time (TAT) of susceptibility testing and culture results, DNA-based ESBL identification would be a valuable surrogate for phenotypic-based methods. Putative ESBL-positive Enterobacteriaceae isolates (n = 330) from clinical specimen were prospectively collected in Bulgaria, Romania and Democratic Republic of Congo and tested in this study. All isolates were assessed for ESBL-production by the E-test method and those giving undetermined ESBL status were re-tested using the combination disk test. A genotypic assay successively combining qPCR detection of blaCTX-M, blaTEM and blaSHV genes with a multiplex pyrosequencing of blaTEM and blaSHV genes was developed in order to detect the most common ESBL-associated TEM and SHV single nucleotides polymorphisms, irrespective of their plasmid and/or chromosomal location. This assay was applied on all Enterobacteriaceae isolates (n = 330). Phenotypic and genotypic results matched in 324/330 (98.2%). Accordingly, real-time PCR combined with multiplex pyrosequencing appears to be a reliable and easy-to-perform assay with high-throughput identification and fast TAT (~5 h).
