Identification of Cryptosporidium spp. oocysts in fecal smears stained with Heidenhain's iron hematoxylin

There is no paucity of methods for diagnosing Cryptosporidium spp. infection. The merits of immunoassays notwithstanding, microscopic identification of Cryptosporidium spp. oocysts in fecal samples remains an important diagnostic procedure. It owes the persistence of its use to such characteristics as dispensing with expensive equipment and kits, requiring only basic laboratory facilities, and having a low probability of false positive results when permanent slides are prepared, which can be re-examined in case of doubt. Cryptosporidium spp. oocysts can be readily identified in fecal smears prepared according to a regressive iron hematoxylin staining technique. The number of steps and their duration, as well as costs, were reduced to a minimum without loss of image quality and permanence of the preparations.

Identification of toxocara canis antigens by Western blot in experimentally infected rabbits

Toxocariasis is a frequent helminthiasis that can cause visceral and ocular damage in humans specially in children. The identification of specific antigens of Toxocara canis is important in order to develop better diagnostic techniques. Ten rabbits were infected orally with a dose of 5000 Toxocara canis embryonated eggs. Rabbits were bled periodically and an ELISA assay was performed to determine levels of specific Toxocara IgG antibodies. ELISA detected antibodies at day 15 after infection. Western blot (WB) assay was performed using excretory/secretory antigens (E/S) of T. canis second stage larvae. Different antigen concentrations were evaluated: 150, 200, 250 and 300 µg/mL. The concentration of 250 µg/mL was retained for analysis. Rabbit sera were diluted 1:100. Secondary antibody was used at a dilution of 1:1000. Results of WB indicated that in the first month after infection specific antibodies against the 200 KDa, 116 KDa, 92 KDa and 35 KDa antigens were detected; antibodies against the 92 KDa, 80 KDa, 66 KDa, 45 KDa, 31 KDa and 28 KDa antigens appeared later. All positive sera in the ELISA test were also positive in WB. Two antigen bands, 92 KDa and 35 KDa, were identified since the beginning and throughout the course of infection. These antigens merit further evaluation as candidates for use in diagnosis.

"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.

Human vaccinia-like virus outbreaks in São Paulo and Goiás States, Brazil: virus detection, isolation and identification

Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the São Paulo and Minas Gerais States and from São Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM) in samples of 49 (66.21%) patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM) of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA) gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus.

Molecular identification of Candida dubliniensis isolated from oral lesions of HIV-positive and HIV-negative patients in São Paulo, Brazil

Candida dubliniensis is a new, recently described species of yeast. This emerging oral pathogen shares many phenotypic and biochemical characteristics with C. albicans, making it hard to differentiate between them, although they are genotypically distinct. In this study, PCR (Polymerase Chain Reaction) was used to investigate the presence of C. dubliniensis in samples in a culture collection, which had been isolated from HIV-positive and HIV-negative patients with oral erythematous candidiasis. From a total of 37 samples previously identified as C. albicans by the classical method, two samples of C. dubliniensis (5.4%) were found through the use of PCR. This study underscores the presence of C. dubliniensis, whose geographical and epidemiological distribution should be more fully investigated.

PCR-based identification of Burkholderia pseudomallei

DNA amplification techniques are being used increasingly in clinical laboratories to confirm the identity of medically important bacteria. A PCR-based identification method has been in use in our centre for 10 years for Burkholderia pseudomallei and was used to confirm the identity of bacteria isolated from cases of melioidosis in Ceará since 2003. This particular method has been used as a reference standard for less discriminatory methods. In this study we evaluated three PCR-based methods of B. pseudomallei identification and used DNA sequencing to resolve discrepancies between PCR-based results and phenotypic identification methods. The established semi-nested PCR protocol for B. pseudomallei 16-23s spacer region produced a consistent negative result for one of our 100 test isolates (BCC #99), but correctly identified all 71 other B. pseudomallei isolates tested. Anomalous sequence variation was detected at the inner, reverse primer binding site for this method. PCR methods were developed for detection of two other B. pseudomallei bacterial metabolic genes. The conventional lpxO PCR protocol had a sensitivity of 0.89 and a specificity of 1.00, while a real-time lpxO protocol performed even better with sensitivity and specificity of 1.00, and 1.00. This method identified all B. pseudomallei isolates including the PCR-negative discrepant isolate. The phaC PCR protocol detected the gene in all B. pseudomallei and all but three B. cepacia isolates, making this method unsuitable for PCR-based identification of B. pseudomallei. This experience with PCR-based B. pseudomallei identification methods indicates that single PCR targets should be used with caution for identification of these bacteria, and need to be interpreted alongside phenotypic and alternative molecular methods such as gene sequencing.

ISO 9001 Quality Management Systems through the Lens of Organizational Culture

Both managers and scholars have convictions about the organizational approaches that best support organizational performance of the respective organizations and its Quality Management Systems. After a literature review of ISO 9001 Quality Management Systems (including the changes introduced by the 2015 edition), Organizational Culture theories are addressed and input from a CEO´s focus group was gathered. The importance of organizational culture for the success of Quality Management Systems and the achievement of the organizational desired results is highlighted. The article advances a proposal to analyze ISO 9001 International Standard through the lens of organizational culture theories identifying a stronger open systems approach (influence of the environment, dynamic perspective, need for survival) of the 2015 ISO 9001 edition when compared with the 2008 one. This provides additional knowledge both to scholars and practitioners for a better understanding of the culture issues that can maximize ISO 9001 Quality Management Systems 2015 edition contributions to organizational enduring success.

Impact of Supplier Management Strategies on the Organizational Performance of ISO 9001 Certified Organizations

Purpose: The purpose of this article is to discuss the impact of Supplier orientation and the resulting Supply Chain Management (SCM) approach, on the organizational performance of ISO 9001 Quality Management Systems certified organizations. Methodology/Approach: Following a literature review, a full structural conceptual model was proposed. An online survey was administered to managers of Portuguese organizations with certified ISO 9001 Quality Management Systems. Descriptive Statistics and Structural Model Equations were used to validate the proposed conceptual model. Findings: There are positive relationships between Organization Strategy and Supplier Orientation, between Supplier Orientation and Stakeholders Satisfaction, and between Stakeholders Satisfaction and Organizational Performance Orientation, supporting ISO 9001:2015. These findings provide insights that supplier orientation, mediated by stakeholder satisfaction, is an essential tool for organizational competitive sustainable advantage. Research Limitation/implication: The analysis was based on managers of ISO 9001 certified organizations perceptions, so additional studies with actual data and longitudinal studies should be useful for further validation. Originality/Value of paper: The importance of the overall organizational ecosystem is highlighted with potential impact on the more than 1 Million ISO 9001 organizations certified worldwide and in their suppliers.

Multilevel model of safety climate for furniture industries

BACKGROUND: Furniture companies can analyze their safety status using quantitative measures. However, the data needed are not always available and the number of accidents is under-reported. Safety climate scales may be an alternative. However, there are no validated Portuguese scales that account for the specific attributes of the furniture sector. OBJECTIVE: The current study aims to develop and validate an instrument that uses a multilevel structure to measure the safety climate of the Portuguese furniture industry. METHODS: The Safety Climate in Wood Industries (SCWI) model was developed and applied to the safety climate analysis using three different scales: organizational, group and individual. A multilevel exploratory factor analysis was performed to analyze the factorial structure. The studied companies’ safety conditions were also analyzed. RESULTS: Different factorial structures were found between and within levels. In general, the results show the presence of a group-level safety climate. The scores of safety climates are directly and positively related to companies’ safety conditions; the organizational scale is the one that best reflects the actual safety conditions. CONCLUSIONS: The SCWI instrument allows for the identification of different safety climates in groups that comprise the same furniture company and it seems to reflect those groups’ safety conditions. The study also demonstrates the need for a multilevel analysis of the studied instrument.

Organizational Culture in Cooperatives: an exploratory approach

In recent years, organizational culture has become one of the common themes of interest of scientific and academic research. Each organization has its own unique cultural identity. Based on the recognition that organizational culture is considered important to an organization’s results, and social economy organizations are concerned with improving managerial practices and results, our objective is to study organizational culture in cooperatives: identifying their organizational culture as a specific type of organization of the social economy, recognized as increasingly important economic agents; and in doing so, explore the usage of a widely known model, the Competing Values Framework (Quinn & Rohrbaugh 1983). Three cooperatives were studied. Their presidents were interviewed, and a questionnaire was applied to cooperative members to obtain demographic and organizational culture data. Differences between the cooperatives’ cultural profiles seem to be consistent with both the circumstances of Portuguese social economy organizations (SEOs), and to the organizations’ uniqueness regarding their trade, focuses, and history. International firm trends were compared with this study’s results, and also appear to be explained by the SEO’s management practices evolution standpoint: lack of structured way of working, and the need to improvise and innovate in order to get things done. The importance of our research is held in the fact that social economy, and the cooperative movement in particular, has a developing importance in the expansion of many economies, the lack of literature on culture in SEOs, and the exploratory usage of a well-known model of management literature in cooperatives.

Genotypic identification of Cryptosporidium spp. isolated from hiv-infected patients and immunocompetent children of São Paulo, Brazil

Cryptosporidium isolates identified in fourteen stool samples, collected from five HIV-infected patients and nine immunocompetent children, living in the Sate of São Paulo, Brazil, were submitted to a molecular analysis using a nested PCR followed of restriction fragment length polymorphism (RFLP), for genetic characterization. The analysis was based on digestion with RsaI restriction enzyme of a DNA fragment amplified from the Cryptosporidium oocyst wall protein (COWP) gene. Based on this analysis, four samples were identified as Cryptosporidium parvum, eight as Cryptosporidium hominis and two presented a profile that correspondedto Cryptosporidium meleagridis when compared to the standards used in the analysis. The use of molecular methods can be helpful to identify source of infections and risk factors related to Cryptosporidium infection in our communities.

Identification of Paracoccidioides brasiliensis in adrenal glands biopsies of two patients with paracoccidioidomycosis and adrenal insufficiency

The authors report two cases of adrenal insufficiency secondary to infiltration of the adrenal glands by Paracoccidioides brasiliensis. The first patient had been treated for a chronic multifocal form of paracoccidiodomycosis 11 years ago. The diagnosis of the mycosis was done simultaneous with that of the adrenal insufficiency in the second patient. In both patients the diagnosis was done by direct visualization of fungus in adrenal biopsies. They were treated with hormonal supplements and itraconazol by 12 and six months, without relapses during the follow-up period.

Generic competencies and practices of managing by competencies: an exploratory study in different organizational contexts

Given the increasingly demanding labor market, today’s professional should act according to certain generic competencies, which allows him to perform as required by the contemporary world. In parallel, human resource management (HRM) current trends, highlight the importance that managing by competencies has in the organizational context, in the sense that it impels a more strategic and integrated HRM. The evidence of the theoretical and practical pertinence of this theme resulted in an exploratory study of a qualitative nature, which findings lead us to conclude that the most valued generic competencies by firms are flexibility, interpersonal relationship, adjustment to change and teamwork. Another finding is that HRM managing by competencies practices is characterized by informality. Managing by competencies is more frequent presence in processes as: recruitment, retention and in the employees’ development plans.

Organizational culture in cooperatives: an exploratory approach

In recent years, organizational culture has become one of the common themes of interest of scientific and academic research. Each organization has its own unique cultural identity. Based on the recognition that organizational culture is considered important to an organization’s results, and social economy organizations are concerned with improving managerial practices and results, our objective is to study organizational culture in cooperatives: identifying their organizational culture as a specific type of organization of the social economy, recognized as increasingly important economic agents; and in doing so, explore the usage of a widely known model, the Competing Values Framework (Quinn & Rohrbaugh 1983). Three cooperatives were studied. Their presidents were interviewed, and a questionnaire was applied to cooperative members to obtain demographic and organizational culture data. Differences between the cooperatives’ cultural profiles seem to be consistent with both the circumstances of Portuguese social economy organizations (SEOs), and to the organizations’ uniqueness regarding their trade, focuses, and history. International firm trends were compared with this study’s results, and also appear to be explained by the SEO’s management practices evolution standpoint: lack of structured way of working, and the need to improvise and innovate in order to get things done. The importance of our research is held in the fact that social economy, and the cooperative movement in particular, has a developing importance in the expansion of many economies, the lack of literature on culture in SEOs, and the exploratory usage of a well-known model of management literature in cooperatives.

An organizational culture approach to performance in information technology micro-firms

This study aims to analyze and compare micro-firms’ organizational culture related to organizational performance. A case study methodology was used based on four firms, competitors among themselves in the Information Technology business, focusing on the years between 2008-2013. Findings pointed out many similarities to larger firms, but some specificities of micro-firms were found and propositions were defined: clan culture predominance is related to best performing micro-firms; the configuration of several culture types seemed to be the most suitable for obtaining good organizational results, provided that they do not focus only on hierarchy and market types of culture; the market culture predominance perception by employees is associated with low job satisfaction; and, after a certain time in business, micro-firms, as do larger companies, seek to standardize and control processes. Recognizing that organizational culture is considered important to firms’ results, this study sheds some light on that important factor for micro-firms.