This research focuses on the lives of migrant mine labourers in the Free State Gold mines of Welkom. After a lengthy absence from home, away from home, some of the migrant mine labourers contact the deadly disease of HIV/AIDS. As they become weaker to work at the mines, some are relegated work at the surface, some are sent to the local hospice whilst some are left to die alone without anyone caring for them. The local community of Welkom regarded these miners as the carriers and transporters of this deadly disease due to their perceived slackness in morality by using the services of the local ladies of the night (commercial sex workers). On the basis of the above painted scenario, the migrant mine labourers were thus subjected to discrimination and stigmatisation by the local community, who regarded them as foreigners even in their country of birth. The research, therefore, aimed to pastorally journey with the infected mine labourers through their trauma of being infected by HIV/AIDS AND of being discriminated and rejected by the community which is supposed to support them in their hour of need and despair. Since the spread of HIV/AIDS is largely through sexual contact, the research, therefore, examined both the community and the church’s attitude towards sex, stigma and discrimination. A participatory observation approach was used and the analysis of the concepts that were at play during the trauma of the infected mine labourers were examined. In this research, the local community was viewed as the fertile ground of hostility against the infected migrant mine labourers. In this regard, the study powered the infected mining community with the challenges posed by HIV/AIDS and therefore sought ways and means of forming a support base for those infected and affected. This was done by breaking the conspiracy of silence around the issue of HIV/AIDS both at the community and the church level. Finally, the study concluded with ways of empowering the pastoral care-giver on how to journey with someone who has been diagnosed with HIV in order to close one’s last chapter of life in honour and dignity. New approaches based on relevant literature and affirmation of God’s power and healing were suggested. / Dissertation (MA(Theol))--University of Pretoria, 2011. / Practical Theology / unrestricted

Orientador: Altair Antoninha Del Bel Cury / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-26T14:52:55Z (GMT). No. of bitstreams: 1 Bertolini_MartinnadeMendoncae_D.pdf: 3303888 bytes, checksum: 17ca780a99a3e63d87bcf4ed94370ffc (MD5) Previous issue date: 2015 / Resumo: O patógeno oportunista Candida albicans e Streptococcus do grupo Mitis formam comunidades complexas em múltiplos sítios da cavidade oral, nos quais o ambiente e a disponibilidade de nutrientes sofrem mudanças constantes. Objetivou-se estudar as características estruturais e de virulência de biofilmes de Candida albicans na presença e ausência de S. oralis crescendo sobre um modelo tri dimensional de mucosa oral humana, em diferentes condições: (1) umidade da superfície mucosa (molhada ou semi seca), (2) disponibilidade de nutrientes (suplementação do meio de cultura com BHI) e (3) morfotipo da hifa (hifa ou pseudo hifa). Para isso foram utilizados modelos tri dimensionais de mucosa oral humana formado por queratinócitos imortalizados (linhagens celulares OKF6-TERT2 ou SCC15) sobre uma matriz colágena com fibroblastos para o crescimento dos biofilmes. Estes foram infectados por Streptococcus oralis 34, e/ou Candida albicans, sendo uma cepa de referência e cepas mutantes para a formação de pseudo-hifas, pela deleção dos genes ndt80 ou tup1. A determinação do biovolume e estrutura do biofilme foram realizadas por microscopia confocal a laser, com os biofilmes sendo corados por imunofluorescência com anticorpo especifico para C. albicans e sonda para Streptococcus. Como determinante de virlência secções de tecido com 5 ?m de espessura foram coradas da mesma maneira anterior ou por hematoxilina e eosina, com o intuito de se detectar a invasão de microorganismos. O dano tecidual também foi mensurado pela liberação de lactato desicrogenase no meio de cultura. Os dados foram avaliados por análises de variâncias (ANOVA) e os procedimentos para comparações múltiplas pareadas por Bonferroni t-test, com ? = 5%. Em condições úmidas C. albicans estendeu hifas longas e entrelaçadas, formando um biofilme de superfície homogênea. Biofilmes mistos apresentaram uma estrutura estratificada, com S. oralis crescendo em contato com a mucosa e a C. albicans cobrindo a superfície bacteriana. Em condições de semi-secas a C. albicans formou densos focos de crescimento localizados a partir dos quais as hifas estenderam-se radialmente para se entrelaçarem com hifas de focos adjacentes. Em biofilmes mistos este fenômeno provocou o acúmulo focal de S. oralis co-localizado com os focos de C. albicans. Embora o biovolume do biofilme de C. albicans tenha sido significativamente maior em condições úmidas (P<0,001), houve uma invasão tecidual mínima em comparação com as condições semi-secas, na qual a barreira epitelial foi completamente destruída. A suplementação do meio de cultura, em condições semi-secas não alterou a arquitetura do biofilme, mas intensificaram o crescimento, o biovolume e a invasão/dano tecidual (P<0,001), proporcionalmente as concentrações testadas. Mutantes para a formação de pseudo-hifas formaram biofilmes defeituosos, nos quais a maioria dos S. oralis estava em contato com a superfície epitelial, abaixo das pseudo-hifas. A presença de S. oralis promoveu invasão e dano tecidual em todas as condições. Conclui-se que a umidade, a disponibilidade de nutrientes, o morfotipo da Candida e a presença de S. oralis afetam fortemente a arquitetura e virulência de biofilmes de C. albicans crescidos sobre nas mucosas / Abstract: The opportunistic pathogen Candida albicans and streptococci of the Mitis group form complex communities in multiple oral sites, where the environment and nutrient availability change constantly. We aimed to study structural and virulence characteristics of Candida albicans biofilms in the presence or absence of S. oralis, growing on a three-dimensional model of human oral mucosa, under different conditions: (1) moisture of mucosal surface (wet or semi dry), (2) nutrient availability (BHI supplementation on culture media) and (3) hyphal morphotype (hyphae or pseudohyphae). For this it was used a three-dimensional model of the human oral mucosa formed by immortalized oral keratinocytes (OKF6-TERT2 or SCC15 cell lines) on a fibroblast-embedded collagenous matrix to grow biofilms. Infections were carried out using Streptococcus oralis 34, a C. albicans reference strain and pseudohyphal mutants with a homozygous deletion of the ndt80, or tup1 gene. Determination of biofilm biovolume and structure was done by confocal scanning laser microscopy with biofilms stained by immunofluorescence using an anti-Candida antibody and a Streptococcus probe. As determinant of virulence, 5-?m-thick tissue sections were stained same way or with hematoxylin and eosin in order to detect invasion of microorganisms. Also tissue damage was measured by lactate dehydrogenase release in the culture media. Statistical analyses were performed using SigmaPlot 12 software at 5% significance level. Data were evaluated by analysis of variance (ANOVA) and when statistical significances were found, all pairwise multiple comparison procedures were performed with Bonferroni t-test, with ? = 5%. Under wet conditions C. albicans extended long intertwined hyphae, forming a homogeneous surface biofilm. Mixed biofilms had a stratified structure, with S. oralis growing in close contact to the mucosa and C. albicans growing on the bacterial surface. Under semi-dry conditions, C. albicans formed localized foci of dense growth from which hyphae extended radially to intertwine with hyphae from adjacent foci. In mixed biofilms this promoted focal growth of S. oralis co-localizing with C. albicans. Although Candida biofilm biovolume was significantly greater under wet conditions (P<0.001), there was minimal tissue invasion compared to semidry conditions where the epithelial barrier was completely destroyed. Supplementing the infection medium with nutrients under semidry conditions did not change the biofilm architecture but intensified focal growth and increased biofilm biovolume and tissue invasion/damage (P<0.001), proportionally to the tested concentrations. Pseudohyphal mutants formed defective mixed biofilms, with most S. oralis in contact with the epithelial surface, below the pseudohyphal mass. Interestingly, the presence of S. oralis promoted fungal invasion and tissue damage under all conditions. Moisture, nutrient availability, hyphal morphotype and presence of S. oralis strongly affect architecture and virulence of mucosal fungal biofilms / Doutorado / Protese Dental / Doutora em Clínica Odontológica

Orientadores: Francisco Hideo Aoki, Ângela Maria de Assis / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T00:12:52Z (GMT). No. of bitstreams: 1 Santos_CristinaRodrigues_M.pdf: 1833064 bytes, checksum: 4087a171a36bf726cff1abdcd0bd4cf8 (MD5) Previous issue date: 2015 / Resumo: O Pneumocystis jirovecii é um fungo ascomiceto que pode causar pneumonia (PPC) em indivíduos imunossuprimidos e eventualmente em imunocompetentes. Os métodos laboratoriais de baixo custo para detecção ainda é a coloração em lâmina. Os métodos moleculares são considerados mais eficientes e tem oferecido alternativas para o entendimento da biologia e da doença causada por esse microrganismo. O objetivo desse estudo foi detectar a presença de Pneumocystis jirovecii em pacientes com doença pulmonar. O método adotado foi a técnica de coloração de lâmina com azul de toluidina para detecção de cistos, PCR para detecção do fragmento mitocondrial pAZ102-H/E, Nested PCR com primers internos pAZ102-X/Y e sequenciamento pelo método de Sanger para detecção de alterações moleculares na região genômica estudada. Foram coletadas 139 amostras, sendo 102 de lavado broncoalveolar (LBA), 7 de escarro, 10 amostras de sangue periférico (fracionadas em hemácias/leucócitos e plasma) e 10 amostras de soro. Nos resultados da coloração pela técnica de azul de toluidina, 2 amostras de escarro e 1 de LBA foram positivas. Através dos métodos moleculares, 6 amostras foram positivas na primeira PCR e 91 amostras foram positivas pela Nested PCR. Os fragmentos pAZ102-X de 51 amostras foram sequenciadas, 18 amostras apresentaram mutação nos seguintes códon/posição 4406/13217, 4419/13257, 4431/13294, 4439/13316, 4440/13321, 4446/13338, 4452/13357, 4456/13367 e 4457/13371. Os pacientes que apresentaram essas alterações tinham um perfil com doenças pulmonares ou fatores de risco associados à infecção por P. jirovecii. Como a detecção de P. jirovecii, diagnóstico e prevenção de PPC são ligadas a história clínica do paciente, os avanços moleculares podem ser um caminho para diagnóstico de PPC, diferenciando essa pneumonia dos casos de colonização por esse fungo / Abstract: Pneumocystis jirovecii is an ascomycete fungus that can cause pneumonia (PCP) in immunocompromised individuals and possibly in immunocompetent. The low cost of laboratory methods for detection is with conventional staining methods. Molecular methods are considered more efficient and have offered alternatives to the understanding of biology and disease caused by this organism. The aim of this study was to detect the target DNA fragment (mtLSUrRNA) of P. jirovecii in patients with lung disease. We compared the staining technique with molecular methods using toluidine blue for microscopy and "in house" DNA extraction, PCR and Nested PCR for molecular detection and sequencing by the Sanger method to detect molecular alterations in the genomic region studied. It was analyzed 139 samples, such as, 102 bronchoalveolar lavage (BAL), 7 sputum, 10 samples of peripheral blood (fractionated in red blood cells / leukocytes and plasma) and 10 serum samples. The results of the toluidine blue staining technique, two sputum samples and 1 BAL were positive. Through molecular methods, 6 samples were positive in the first PCR and 91 samples were positive by Nested PCR. The pAZ102-X fragments of 51 samples were sequenced, 18 samples had mutations in codon/position 4406/13217, 4419/13257, 4431/13294, 4439/13316, 4440/13321, 4446/13338, 4452/13357, 4456 / 13367 and 4457/13371. Patients who presented these alterations had a profile with lung diseases or risk factors associated with infection by P. jirovecii. As the detection of P. jirovecii, diagnosis and prevention of PCP are linked to clinical history, molecular advances can be a way for diagnosis of PCP, differentiating this pneumonia cases of colonization by the fungus. OBSERVAÇÃOPor favor manter essa palavra em itálico: Pneumocystis jirovecii / Mestrado / Clinica Medica / Mestra em Ciências

Background: Microsporidia are obligate intracellular opportunistic fungi that cause significant pathology in immunocompromised hosts. However, 11 percent of immunocompetent individuals in the general population are microsporidia-seropositive, indicating that severe immune suppression may not be a prerequisite for infection. Encephalitozoon intestinalis is transmitted in contaminated water and initially infects gastro-intestinal enterocytes, leading to diarrheal disease. This organism can also disseminate to many other organs. A recent report suggests that microsporidia can establish persistent infections, which anti-fungal treatment does not eradicate. Like other intracellular pathogens, microsporidia infection stresses the host cell and infected individuals have elevated hydrogen peroxide and free radical levels. Findings. As oxidative stress can lead to DNA damage, we hypothesized that E. intestinalis-infection would increase host cell nuclear mutation rate. Embryo fibroblasts from Big Blue§ssup§TM§esup§ transgenic mice were E. intestinalis-infected and host nuclear mutation frequency was determined by selection of temperature-sensitive c-II gene mutant λ phage. The host mutation frequency in E. intestinalis-infected cultures was 2.5-fold higher than that observed in either mock-infected cells or cells infected with UV-inactivated E. intestinalis spores. Conclusions: These data provide the first evidence that microsporidia infection can directly increase host cellular mutation frequency. Additionally, some event in the microsporidia developmental cycle between host cell attachment and parasitophorous vacuole formation is required for the observed effect. As there is considerable evidence linking infection with other intracellular pathogens and cancer, future studies to dissect the mechanism by which E. intestinalis infection increases host mutation frequency are warranted.

Background: Microsporidia are obligate intracellular opportunistic fungi that cause significant pathology in immunocompromised hosts. However, 11 percent of immunocompetent individuals in the general population are microsporidia-seropositive, indicating that severe immune suppression may not be a prerequisite for infection. Encephalitozoon intestinalis is transmitted in contaminated water and initially infects gastro-intestinal enterocytes, leading to diarrheal disease. This organism can also disseminate to many other organs. A recent report suggests that microsporidia can establish persistent infections, which anti-fungal treatment does not eradicate. Like other intracellular pathogens, microsporidia infection stresses the host cell and infected individuals have elevated hydrogen peroxide and free radical levels. Findings. As oxidative stress can lead to DNA damage, we hypothesized that E. intestinalis-infection would increase host cell nuclear mutation rate. Embryo fibroblasts from Big Blue§ssup§TM§esup§ transgenic mice were E. intestinalis-infected and host nuclear mutation frequency was determined by selection of temperature-sensitive c-II gene mutant λ phage. The host mutation frequency in E. intestinalis-infected cultures was 2.5-fold higher than that observed in either mock-infected cells or cells infected with UV-inactivated E. intestinalis spores. Conclusions: These data provide the first evidence that microsporidia infection can directly increase host cellular mutation frequency. Additionally, some event in the microsporidia developmental cycle between host cell attachment and parasitophorous vacuole formation is required for the observed effect. As there is considerable evidence linking infection with other intracellular pathogens and cancer, future studies to dissect the mechanism by which E. intestinalis infection increases host mutation frequency are warranted.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 4 uppsatser.

Submitted by Luanna Matias (lua_matias@yahoo.com.br) on 2015-03-11T16:13:39Z No. of bitstreams: 2 Dissertação - Solange Moreira da Silva - 2012.pdf: 2785230 bytes, checksum: 7aaa8506c7565237cf88ab856e88a595 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luanna Matias (lua_matias@yahoo.com.br) on 2015-03-11T20:03:28Z (GMT) No. of bitstreams: 2 Dissertação - Solange Moreira da Silva - 2012.pdf: 2785230 bytes, checksum: 7aaa8506c7565237cf88ab856e88a595 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-03-11T20:03:28Z (GMT). No. of bitstreams: 2 Dissertação - Solange Moreira da Silva - 2012.pdf: 2785230 bytes, checksum: 7aaa8506c7565237cf88ab856e88a595 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2012-12-12 / Oral manifestations are common in Human Immunodeficiency Virus (HIV) infected individuals and are present at any moment of this morbidity. They are caused by opportunistic agents like fungus, virus or bacteria and neoplasms occur sometimes. The objective of this study was to develop a systematic literature review to know about the prevalence for oral manifestations related to HIV infection and to verify their occurrence in patients infected with HIV, followed at Sistema Único de Saúde, associated factors and side effects related to antirretroviral therapy. Initially was developed a systematic literature review to answer about oral manifestation prevalence and their meaning in HIV infection. Simultaneously an observational study was initiated including 201 HIV infected patients regulated by the Sistema Único de Saúde. These patients were currently attended at the Ambulatory of Infectology from Hospital das Clínicas of Universidade Federal de Goiás. All the participants were submitted to an oral examination which permitted a presumptive diagnostic based on international criteria of The EC Clearinghouse on Oral manifestations for HIV infection recommended by The World Health Organization. They also answered to a structured formulary e data were collected from the medical records. Characteristics lesions were photographed. The systematic literature review identified 337 papers. Only 20 were finally included after Relevance Tests I and II. These papers revealed a prevalence for oral manifestations related to HIV ranging from 19, 5% to 85,0%. Candidosis and oral hairy leucoplakia were the most prevalent lesions and they have been considered as immunologic markers for disease progression and therapeutic failure. The statistical analyse were descriptive and univariate logistic regression surveyed associations between oral manifestations related to human immunedeficiency virus infection and sociodemographic datas, CD4+ counting, time of disease, use of antiretroviral therapy, prosthesis usage and presence of carie lesion. Fisher’s exact test was applied to comparisons. At the observational study we found a low prevalence for these lesions with significant difference between groups HIV+ and AIDS. There were 5,40% of periodontitis related to HIV infection in the Group I and 11,00% in the Goup II. Neoplasms were not found. The most common side effect for antiretroviral therapy was dry mouth (7,60% at the HIV Group and 11,00% at the AIDS Group). There was statistical significant association between oral lesion occurrence and carie presence (AIDS Group) or use of prosthesis and age (HIV Group). From the systematic literature review we concluded that oral manifestations are common in HIV+/AIDS patients and candidosis and oral hairy leucoplakia are immunologic markers for the disease progression and Abstract xx therapeutic failure. From the observational study we concluded that the prevalence verified was relatively short. Periodontal diseases were the most common lesions. Age and prosthesis use were associated to oral manifestations occurrence at the HIV Group and carie lesions also influenced at the AIDS Group. / As manifestações bucais em indivíduos infectados pelo Virus da Imunodeficiência Humana (HIV) são comuns e estão presentes em algum momento da infecção. Causadas por germes oportunistas como fungos, bactérias e vírus, podem resultar até mesmo em lesões malignas. Este estudo objetivou realizar uma revisão sistemática da literatura científica para investigar a prevalência das manifestações bucais relacionadas ao HIV em pacientes infectados por este vírus e verificar a ocorrência das mesmas em pacientes atendidos pelo Sistema Único de Saúde, possíveis fatores associados e efeitos adversos bucais relacionados ao uso de antirretrovirais. Inicialmente uma revisão sistemática da literatura verificou a prevalência destas lesões e sua importância no curso da doença. Simultaneamente foi desenvolvido um estudo observacional incluindo 201 pacientes infectados pelo HIV, sendo 92 pertencentes ao Grupo I (HIV+) e 109 pertencentes ao Grupo II (AIDS), regulados pelo Sistema Único de Saúde, atendidos no Ambulatório de Infectologia do Hospital das Clínicas da Universidade Federal de Goiás. Todos os participantes foram submetidos a exame da cavidade bucal e o diagnóstico presuntivo das manifestações bucais foi baseado em critérios internacionais do European Community – Clearinghouse recomendado pela Organização Mundial de Saúde. A seguir, todos responderam a um formuário estruturado do estudo e dados foram coletados dos prontuários médicos. Lesões consideradas relevantes foram fotografadas. Na revisão sistemática, dos 337 artigos científicos identificados, apenas 20 atenderam aos critérios de inclusão retratando uma variação na prevalência de lesões orais entre 19,50% a 85,00%. Nesta revisão, a candidose e a leucoplasia pilosa foram as mais lesões bucais mais prevalentes, consideradas marcadores da progressão da doença e de falência terapêutica. No estudo observacional a análise estatística foi descritiva e a verificação de associações entre a ocorrência de manifestações bucais relacionados à infecção pelo vírus da imunodeficiência humana e dados sóciodemográficos, contagem de CD4+, tempo de doença, uso de terapia antirretroviral, uso de prótese e presença de lesão de cárie foi realizada por meio de regressão logística univariada. Para comparações foi utilizado o teste exato de Fisher. No estudo observacional, foi encontrada uma prevalência de 19,60% para o grupo I e de 33,00% para o Grupo II para as manifestações bucais relacionadas ao HIV. Verificou-se diferença estatisticamente significante entre os dois grupos. Houve uma prevalência de 5,40% de periodontite relacionada ao HIV no Grupo I e 11,00% no Grupo II. Não foram encontradas lesões malignas. O efeito adverso bucal relacionado ao uso de antirretroviral mais comum nos dois grupos foi xerostomia (7,60% no Grupo HIV e 11,00% no Grupo AIDS). Houve associação estatisticamente significante entre a ocorrência de lesões bucais e a presença de cárie (Grupo AIDS) e idade e uso de prótese (Grupo HIV). Da revisão sistemática da literatura conclui-se que estas manifestações são comuns nos pacientes HIV+/AIDS e que candidose e leucoplasia pilosa bucal são marcadores imunológicos da doença e de falência terapêutica. Do estudo observacional concluímos que a prevalência de manifestações bucais relacionadas ao HIV foi relativamente baixa. As doenças periodontais relacionadas ao HIV foram as lesões mais prevalentes. Idade e uso de prótese foram fatores influenciadores na ocorrência dessas manifestações bucais no Grupo HIV. No Grupo AIDS, a presença de lesão de cárie influenciou na ocorrência dessas manifestações.

Chronic granulomatous disease (CGD) is associated with defective function of the NADPH-oxidase system in conjunction with phagocytic defects which leads to granuloma formation and serious infectious complications. This is often associated with significant morbidity and mortality. The association of defective phagocyte function with other coincidental immune defects is unknown. Defects in innate pathways seen with CGD, including complement systems, and toll-like and dectin receptor pathways, have not been described before. We present the case of a 2-year old male patient hospitalized with recurrent pneumonia, a non-healing skin ulcer, necrotizing lung granulomas, and epididymo-orchitis. Defective neutrophil chemiluminescence was detected by dihydrorhodamine (DHR) testing. Further evaluation demonstrated characteristic molecular mutations of CYBB consistent with CGD. Immune evaluation demonstrated polyclonal hyperglobulinemia, but a greatly reduced mannose binding lectin (MBL) level. Six biallelic polymorphisms in MBL gene and its promoter were analyzed using Light Cycler™ Real-time PCR assay. The LXPA/LYPB haplotype of MBL was detected in our patient; the latter is the defective haplotype associated with low MBL levels. Due to the implications for innate immunity and the protection against bacterial, viral, and fungal infections provided by MBL, a deficiency of this protein may have disastrous consequences on the long term outcomes of CGD. MBL deficiency can also complicate other disorders affecting the immune system, significantly increasing the risk of infection in such patients. Further studies looking at the frequency and implications of MBL deficiency in CGD are needed. © 2012 Bentham Science Publishers.

Microbial interactions represent an understudied facet of human health and disease. In this study, the interactions that occur between Chlamydia trachomatis and the opportunistic fungal pathogen, Candida albicans were investigated. Candida albicans is a common component of the oral and vaginal microbiota responsible for thrush and vaginal yeast infections. Normally, Candida exist in the body as yeast. However, disruptions to the microbiota create conditions that allow expanded growth of Candida, conversion to the hyphal form, and tissue invasion. Previous studies have shown that a myriad of outcomes can occur when Candida albicans interacts with pathogenic bacteria. To determine if C. trachomatis physically interacts with C. albicans, we incubated chlamydial elementary bodies (EB) in medium alone or with C. albicans yeast or hyphal forms for 1 h. Following incubation, the samples were formaldehyde-fixed and processed for immunofluorescence assays using anti-chlamydial MOMP or anti- chlamydial LPS antibodies. Replicate samples were replenished with culture medium and incubated at 35°C for 0-120 h prior to fixation for immunofluorescence analysis or collection for EB infectivity assays. Data from this study indicates that both C. trachomatis serovar E and C. muridarum EB bind to C. albicans yeast and hyphal forms. This interaction was not blocked by pre-incubation of EB with the Candida cell wall components, mannan or β-glucans, suggesting that EB interact with a Candida cell wall protein or other structure. Bound EB remained attached to C. albicans for a minimum of 5 days (120 h). Infectivity assays demonstrated that EB bound to C. albicans are infectious immediately following binding (0h). However, once bound to C. albicans, EB infectivity decreased at a faster rate than EB in medium alone. At 6h post binding, 40% of EB incubated in medium alone remained infectious compared to only 16% of EB bound to C. albicans. Likewise, pre-incubation of EB with laminarin, a soluble preparation of β-glucan, alone or in combination with other fungal cell wall components significantly decreases chlamydial infectivity in HeLa cells. These data indicate that interactions between EB and C. albicans inhibit chlamydial infectivity, possibly by physically blocking EB interactions with host cell receptors.

When patients develop new-onset infections after hospital admission, the origin of the infection is typically assumed to be nosocomial; however, ambulances are potentially unexplored reservoirs for emerging pathogens. This study seeks to identify the scope of bacterial contamination in rural East Tennessee ambulances. Though universal precautions and cleaning procedures aim to reduce the spread of infectious diseases to provider and patient, little is known about the bacterial microbiome of ambulances. To the best of our knowledge, this is the first study of its kind to be performed in the state of Tennessee and the first since the introduction of UVGI units as an ambulance-based COVID-19 infection control measure. Our dissemination of post-pandemic findings may impact ambulance sanitation measures and will add to the national and global knowledge pertaining to the microbiome of emergency medical patient transport systems. Ambulances in East Tennessee were sampled using environmental sampling contact plates. At least one active ambulance unit for each EMS service underwent sampling. Three samples were obtained from each of three areas: the floor of the ambulance transport area, the rear door panel inside the transport area and stretcher. The plates were then incubated at 30-35C for 48 hours. Colony counts were manually performed before the plates were shipped for species identification via MALDI-TOF DNA analysis by MIDI laboratories (Newark, DE). One plate from each ambulance door and stretcher was sent for bacterial identification. Only one sample returned free of growth. All floor samples, several stretcher samples, and three door samples presented vast growth with colonies too numerous to count. The results from bacterial identification showed all flora were human commensal flora or environmental flora. The flora found on ambulance doors with opportunistic capabilities are as follows: Staphylococcus hominis, Staphylococcus epidermidis, Enterobacter cloacae, Enterobacter xinagfangensis, Bacillus cereus, Klebsiella oxytoca, and Bacillus subtilis; and the flora found on the stretchers with opportunistic capabilities are as follows: Staphylococcus haemolyticus, Staphylococcus epidermidis, Staphylococcus cohnii ssp urealyticus, Bacillus cereus, Corynebaccterium mucifaciens, Staphylococcus pettenkoferi, Klebsiella oxytoca, Staphylococcus capitis, Bacillus subtillis, and Staphylococcus caprae. In this era of increasing antibiotic resistance, it is concerning that several microbes with pathogenicity were found, including species that often confer the spread of resistance such as Klebsiella oxytoca and Enterobacter cloacae. Overall, the finding of numerous diverse colonies does not support adequate sanitation of the ambulances. Further study is required to identify the most effective sanitation methods, and further metagenomic study is needed to explore the presence of genes that facilitate the spread of microbial resistance.