A Bacillus thuringiensis isolation method utilizing a novel stain, low selection and high throughput produced atypical results

BACKGROUND: Bacillus thuringiensis is a bacterium known for producing protein crystals with insecticidal properties. These toxins are widely sought after for controlling agricultural pests due to both their specificity and their applicability in transgenic plants. There is great interest in isolating strains with improved or novel toxin characteristics, however isolating B. thuringiensis from the environment is time consuming and yields relatively few isolates of interest. New approaches to B. thuringiensis isolation have been, and continue to be sought. In this report, candidate B. thuringiensis isolates were recovered from environmental samples using a combination of a novel stain, high throughput and reduced selection. Isolates were further characterized by SDS-PAGE, light microscopy, PCR, probe hybridization, and with selected isolates, DNA sequencing, bioassay or Electron Microscopy. RESULTS: Based on SDS-PAGE patterns and the presence of cry genes or a crystal, 79 candidate, non-clonal isolates of B. thuringiensis were identified from 84 samples and over 10,000 colonies. Although only 16/79 (20%) of the isolates showed DNA homology by Probe Hybridization or PCR to common cry genes, initial characterization revealed a surprisingly rich library that included a putative nematocidal gene, a novel filamentous structure associated with a crystal, a spore with spikes originating from a very small parasporal body and isolates with unusually small crystals. When compared to reports of other screens, this screen was also atypical in that only 3/79 isolates (3.8%) produced a bipyramidal crystal and 24/79 (30%) of the isolates' spores possessed an attached, dark-staining body. CONCLUSION: Results suggest that the screening methodology adopted in this study might deliver a vastly richer and potentially more useful library of B. thuringiensis isolates as compared to that obtained with commonly reported methodologies, and that by extension, methodologies fundamentally different from current methods should also be explored

In-house polymerase chain reaction for affordable and sustainable Chlamydia trachomatis detection in Trinidad and Tobago

Objectives: To provide a preliminary assessment of in-house polymerase chain reaction (PCR) as an alternative to the more costly commercial test for detection of asymptomatic infection by Chlamydia trachomatis and to provide much needed demographic data on infection indicators within the Trinidad and Tobago public health care system. Methods: An inexpensive in-house nested-PCR with an Internal Amplification Control was used to detect C. trachomatis and Neisseria gonorrhoeae in urine samples collected from 273 apparently healthy, pregnant women from March-September 2004 in Trinidad, West Indies. Demographic information on participants was collected and subjected to statistical analyses. Results: C. trachomatis was detected in 57/273 (21%) samples, of which 5 (2%) were also positive for N. gonorrhoeae. Infection correlated well with certain demographic parameters, with the highest incidence of C. trachomatis infection found among pregnant women that were single or of African descent. Conclusions: Given the lack of commercial tests in Trinidad, in-house PCR is an inexpensive alternative that can be used to detect asymptomatic infections of C. trachomatis and to provide demographic information needed for interventions by the public health care system

Comparison of polymerase chain reaction and bacterial culture for Salmonella detection in the Muscovy duck in Trinidad and Tobago

Objectives: The purpose of this study was to investigate the presence and serovar identity of Salmonella, at the national level, in farmed Muscovy ducks (Cairina moschata) in Trinidad and Tobago, and to compare the relative benefits of bacterial culture to those of polymerase chain reaction (PCR) for use in the routine detection and surveillance of Salmonella in these ducks. Methods: From March-September 2003, 110 fecal samples were collected from 82 farms across the islands of Trinidad and Tobago. Salmonella was isolated from fresh and frozen samples and the serotype of each was determined through bacterial culture. An in-house, nested PCR that detects all pathogenic Salmonella species was utilized in analyzing the samples. Results: Five samples were positive for Salmonella by bacterial culture, whereas 44 were positive by the nested PCR. Serovars isolated were Kiambu, Orion, Uganda, and two isolates from Group E1 whose H antigens could not be fully characterized. Of the samples, 87 (79%) gave equivalent PCR results for both enrichment broths-28 were positive for both and 59 were negative for both). However, 16 samples were positive for one broth, but not for the other, with the majority (14 of the 16) resulting positive for Selenite broth. PCR results for seven samples were inconclusive due to ambiguous band size or multiple bands near the expected band size. Conclusions: In Trinidad and Tobago, the Muscovy duck does not appear to be a significant source of S. typhimurium or S. enteritidis, but it does harbor other Salmonella species. In-house, nested PCR represents a simple, relatively inexpensive and potentially more sensitive method than bacterial culture for the routine surveillance of pathogenic Salmonella in the Muscovy duck

Perceptions of Collegial and Uncollegial Behaviors After a University Consolidation: A Quantitative and Qualitative Analysis of How Faculty Viewed Members of Their New Academic Units

Much has been written about collegiality in academe, most notably by Cipriano (2011), Buller (2006, 2012), and Cipriano and Buller (2012, 2017), Flaherty (2013). Concomitantly, awareness has increased about instances of abusive supervision (Gere, 2020), incivility (Andersson & Pearson, 1999), microaggressions (Sue & Rivera, 2011) bullying and mobbing (i.e., group bullying) in the workplace and in higher education (Cowan, 2009), Duffy (2009), Lutgen-Sandvik (2006), Lutgen-Sandvik and Tracy (2012), Heeman (2007), Lutgen-Sandvik & McDermott (2011), and Taylor (2012). Instances of incivilities have continued to be a concern as evident in the journal article in Nature titled: “Astronomers victimized colleagues—and put historic Swedish department in turmoil,” in which Witze (2021) reported that two high ranking faculty members (one male, one female) were investigated and found responsible for bullying at Lund University. Bullying in the academy is not confined to one country, one discipline, or one gender. Based on a review of the literature on university consolidations and on collegiality in academic settings, the research team found that there was a gap in the literature regarding how participants of a university consolidation (sometimes called mergers) perceive their environment in a departmental (or equivalent unit level), especially a “new” unit that has been formed because of the consolidation of two or more units from previously existing (legacy) institutions. Cipriano and Buller (2012) have used the CAM (Collegiality Assessment Matrix) and/or the Self-Assessment Matrix of Collegiality (SAM), proprietary instruments, to measure the “collegiality” of individuals in academic departments. However, there has not been an assessment of collegiality from a “departmental or equivalent unit” level perspective. This study, therefore, addresses this “gap” in the research. Moreover, this study expands the discussion of collegiality to include the identification of perceived uncollegial (conflict) behaviors of incivility, microaggressions (such as misogynistic statements), bullying, and mobbing

IntelliBeeHive: An Automated Honey Bee, Pollen, and Varroa Destructor Monitoring System

—study, we developed a honey bee monitoring system that aims to enhance our understanding of Colony Collapse Disorder, honey bee behavior, population decline, and overall hive health. The system is positioned at the hive entrance providing real-time data, enabling beekeepers to closely monitor the hive’s activity and health through an account-based website. Using machine learning, our monitoring system can accurately track honey bees, monitor pollen gathering activity, and detect Varroa mites, all without causing any disruption to the honey bees. Moreover, we have ensured that the development of this monitoring system utilizes cost-effective technology, making it accessible to apiaries of various scales, including hobbyists, commercial beekeeping businesses, and researchers. The inference models used to detect honey bees, pollen, and mites are based on the YOLOv7-tiny architecture trained with our own data. The F1-score for honey bee model recognition is 0.95 and the precision and recall value is 0.981. For our pollen and mite object detection model F1-score is 0.95 and the precision and recall value is 0.821 for pollen and 0.996 for ”mite”. The overall performance of our IntelliBeeHive system demonstrates its effectiveness in monitoring the honey bee’s activity, achieving an accuracy of 96.28% in tracking and our pollen model achieved a F1-score of 0.831

IntelliBeeHive: An Automated Honey Bee, Pollen, and Varroa Destructor Monitoring System

Utilizing computer vision and the latest technological advancements, in this study, we developed a honey bee monitoring system that aims to enhance our understanding of Colony Collapse Disorder, honey bee behavior, population decline, and overall hive health. The system is positioned at the hive entrance providing real-time data, enabling beekeepers to closely monitor the hive's activity and health through an account-based website. Using machine learning, our monitoring system can accurately track honey bees, monitor pollen-gathering activity, and detect Varroa mites, all without causing any disruption to the honey bees. Moreover, we have ensured that the development of this monitoring system utilizes cost-effective technology, making it accessible to apiaries of various scales, including hobbyists, commercial beekeeping businesses, and researchers. The inference models used to detect honey bees, pollen, and mites are based on the YOLOv7-tiny architecture trained with our own data. The F1-score for honey bee model recognition is 0.95 and the precision and recall value is 0.981. For our pollen and mite object detection model F1-score is 0.95 and the precision and recall value is 0.821 for pollen and 0.996 for "mite". The overall performance of our IntelliBeeHive system demonstrates its effectiveness in monitoring the honey bee's activity, achieving an accuracy of 96.28 % in tracking and our pollen model achieved a F1-score of 0.831

An Exploratory Study of Methicillin-Resistant Staphylococcus aureus and SCCmec Elements Obtained from a Community Setting Along the Texas Border with Mexico

An exploratory study of methicillin-resistant Staphylococcus aureus (MRSA) and SCCmec elements in bacteria along the Mexican border of south Texas was performed. Between September and December of 2008, 375 swabs of anterior nares were self-collected by students attending the University of Texas-Pan American (UTPA) and cultured for MRSA. Fifty seven bacterial isolates were kept for further analysis that included suspected MRSA and other SCCmec-containing bacteria. Isolates were examined for the presence of nuc, mecA, lukS-PV, and spa genes using PCR. SCCmec and spa typing were also performed. Seven S. aureus isolates were found of which six were classified as MRSA. SCCmec typing showed five of the six MRSA strains to be type IV, while one MRSA strain, and most of the non-S. aureus strains, were untypeable, producing results that were indicative of mixed SCCmec types. Five of the six MRSA strains contained known spa types (two of which corresponded to USA300 and one to USA600), while one strain had a novel spa type. Only one isolate, a USA300 MRSA, was positive for lukS-PV. Easy access by the Texas border community to antibiotics in Mexico without a prescription, and the strong partition in SCCmec types between MRSA and non-S. aureus bacteria suggest that this border region of Texas may be uniquely suited for the study of emerging SCCmec types, their horizontal transfer, and perhaps other aspects of antibiotic resistance in bacteria

Improving STEM Education in Research: Preliminary Report on the Development of a Computer-Assisted Student-Mentor Research Community

Research education in STEM disciplines currently suffers from 1) The inability to feasibly collect highly detailed data on both the student’s and mentor’s activities; 2) The lack of tools to assist students and mentors in organizing and managing their research activities and environments; and 3) The inability to correlate a student’s assessment results with their actual research activities. Together these three problems act to impede both the improvement and educational quality of student research experiences. We propose a computer-assisted student-mentor research community as a solution to these problems. Within this community setting, students and their mentors are provided tools to make their work easier, much like a word processor makes writing a letter easier. Through their use of these tools, details of student-mentor activities are automatically recorded in a relational database, without burdening users with the responsibility of archiving data. Equally important, student assessments of outcome can be directly related to student activity, allowing educators to identify practices resulting in successful research experiences. Community tools also facilitate the use of labor-intensive teaching laboratories involving real inquiry-based research. The community structure has the added benefit of allowing students to see, communicate and interact more freely with other students and their projects, thus enriching the student’s research experience. We provide herein a preliminary report on the development and testing of a prototype, student-mentor research community, and present its tools, an assessment of student interest in participating in the community, and discuss its further development into a nationally-available student-mentor research community

A nested-PCR with an Internal Amplification Control for the detection and differentiation of Bartonella henselae and B. clarridgeiae: An examination of cats in Trinidad

BACKGROUND: Bartonella species are bacterial blood parasites of animals capable of causing disease in both animals and man. Cat-Scratch Disease (CSD) in humans is caused mainly by Bartonella henselae and is acquired from the cat, which serves as a reservoir for the bacteria. A second species, B. clarridgeiae is also implicated in the disease. Diagnosis of Bartonellosis by culture requires a week or more of incubation on enriched media containing blood, and recovery is often complicated by faster growing contaminating bacteria and fungi. PCR has been explored as an alternative to culture for both the detection and species identification of Bartonella, however sensitivity problems have been reported and false negative reactions due to blood inhibitors have not generally been addressed in test design. METHODS: A novel, nested-PCR was designed for the detection of Bartonella henselae and B. clarridgeiae based on the strategy of targeting species-specific size differences in the 16S-23S rDNA intergenic regions. An Internal Amplification Control was used for detecting PCR inhibition. The nested-PCR was utilized in a study on 103 blood samples from pet and stray cats in Trinidad. RESULTS: None of the samples were positive by primary PCR, but the Nested-PCR detected Bartonella in 32/103 (31%) cats where 16 were infected with only B. henselae, 13 with only B. clarridgeiae and 3 with both species. Of 22 stray cats housed at an animal shelter, 13 (59%) were positive for either or both species, supporting the reported increased incidence of Bartonella among feral cats. CONCLUSION: The usefulness of a single PCR for the detection of Bartonella henselae and B. clarridgeiae in the blood of cats is questionable. A nested-PCR offers increased sensitivity over a primary PCR and should be evaluated with currently used methods for the routine detection and speciation of Bartonella henselae and B. clarridgeiae. In Trinidad, B. henselae and B. clarridgeiae are the predominant species in cats and infection appears highest with stray cats, however B. clarridgeiae may be present at levels similar to that of B. henselae in the pet population