Culture-independent characterization of bacteria and fungi in a poultry bioaerosol using pyrosequencing: a new approach.

Work in animal production facilities often results in exposure to organic dusts. Previous studies have documented decreases in pulmonary function and lung inflammation among workers exposed to organic dust in the poultry industry. Bacteria and fungi have been reported as components of the organic dust produced in poultry facilities. To date, little is known about the diversity and concentration of bacteria and fungi inside poultry buildings. All previous investigations have utilized culture-based methods for analysis that identify only biota cultured on selected media. The bacterial tag-encoded flexible (FLX) amplicon pyrosequencing (bTEFAP) and fungal tag-encoded flexible (FLX) amplicon pyrosequencing (fTEFAP) are modern and comprehensive approaches for determining biodiversity of microorganisms and have not previously been used to provide characterization of exposure to microorganisms in an occupational environment. This article illustrates the potential application of this novel technique in occupational exposure assessment as well as other settings. An 8-hr area sample was collected using an Institute of Medicine inhalable sampler attached to a mannequin in a poultry confinement building. The sample was analyzed using bTEFAP and fTEFAP. Of the bacteria and fungi detected, 116 and 39 genera were identified, respectively. Among bacteria, Staphylococcus cohnii was present in the highest proportion (23%). The total inhalable bacteria concentration was estimated to be 7503 cells/m³. Among the fungi identified, Sagenomella sclerotialis was present in the highest proportion (37%). Aspergillus ochraceus and Penicillium janthinellum were also present in high proportions. The total inhalable fungi concentration was estimated to be 1810 cells/m³. These estimates are lower than what has been reported by others using standard epifluorescence microscope methods. However, no study has used non-culture-based techniques, such as bTEFAP and fTEFAP, to evaluate bacteria and fungi in the inhalable fraction of a bioaerosol in a broiler production environment. Furthermore, the impact of this bTEFAP and fTEFAP technology has yet to be realized by the scientific community dedicated to evaluating occupational and environmental bioaerosol exposure.

Soluble complement receptor 1 (sCR1) protects against experimental autoimmune myasthenia gravis.

The loss of muscle function seen in myasthenia gravis and in the animal model of the disease, experimental autoimmune myasthenia gravis (EAMG) is in part due to the activation of complement by anti-acetylcholine receptor (AChR) antibodies at the motor end-plate. In this study we describe the effects of a soluble recombinant form of human complement receptor 1 (sCR1) on the development of clinical disease and receptor loss in EAMG induced passively by administration of anti-AChR antibodies. Daily intraperitoneal injection of sCR1 significantly reduced the weight loss and severity of clinical symptoms seen and allowed treated animals to recover normal muscle function. These data suggest that sCR1 could provide a useful additional therapeutic agent in myasthenia.

Effect of continuous complement inhibition using soluble complement receptor type 1 on survival of pig-to-primate cardiac xenografts.

A single bolus of soluble complement (C) receptor type 1 (sCR1, TP-10) has been shown to delay hyperacute rejection (HAR) of porcine cardiac xenografts (Xgs) by primate recipients. In these recipients, C activity slowly returned and C deposition was noted in the Xgs at rejection. To evaluate the effect of sustained C inhibition using sCR1 on HAR, two additional cynomolgus monkeys received porcine cardiac Xgs and a continuous infusion of sCR1. In the first recipient, Xgs survival was 5 days (120+ hr), whereas in the second, Xg survival was 7 days (168+ hr). Serial biopsies of the Xgs were remarkable for an increasing cellular infiltrate composed predominantly of neutrophils and macrophages, and the development of edema, hemorrhage, and myocyte necrosis. These findings suggest that once C-mediated HAR has been inhibited, infiltration of the Xg by these cells may lead to accelerated acute rejection, which is an additional barrier to successful longer term Xg survival.

Inhibition of complement, evoked antibody, and cellular response prevents rejection of pig-to-primate cardiac xenografts.

Complement (C) inhibition alone using a recombinant soluble form of complement receptor type 1 (sCR1) prevents hyperacute rejection but not subsequent irreversible accelerated acute rejection of discordant pig-to-cynomolgus monkey cardiac xenografts, which occurs within 1 week. To inhibit accelerated acute rejection, which is associated with a rise in serum xenoreactive antibody (Ab) and a cellular infiltrate, triple therapy with standard immunosuppressive agents (cyclosporine, cyclophosphamide, and steroids [CCS]) was combined with continuous C inhibition using sCR1. Each of two monkeys that received sCR1 + CCS showed minimal evidence of rejection when killed on days 21 and 32 in comparison to a monkey that received sCR1 + subtherapeutic CCS (rejected at 11 days) and a control that received CCS alone (rejected at 38 min). Prolonged xenograft survival was associated with low Ab levels and a minimal cellular infiltrate, suggesting that combined inhibition of C, xenoreactive Ab responses, and cellular immunity may be a useful approach in overcoming the immune barriers to discordant xenotransplantation.

The effect of soluble complement receptor type 1 on hyperacute rejection of porcine xenografts.

The use of xenografts (Xgs) from distantly related species to relieve the increasing shortage of organs for clinical transplantation is prevented by the occurrence of hyperacute rejection (HAR). This process, in which C activation plays a central role, cannot be inhibited with currently available immunosuppressants. In two clinically relevant xenotransplantation models, this study evaluated the effect of C inhibition using recombinant soluble complement receptor type 1 (sCR1) on HAR. In an ex vivo model in which porcine cardiac Xgs were perfused with human blood, cardiac function ceased within 34 min when the perfusate blood was untreated (n = 3). When the perfusate blood was treated with sCR1 (300 micrograms/ml), cardiac Xg function was maintained for up to 4 hr (n = 3). Immunohistologic examination of these Xgs demonstrated deposition of C3b/iC3b and C3d in Xgs perfused with untreated human blood but only C3d deposition in those Xgs perfused with sCR1-treated human blood. These findings are consistent with the cofactor activity of sCR1 for factor I-mediated degradation of deposited C3b/iC3b to C3d. Treatment with sCR1 also prevented the histopathologic changes of HAR observed when untreated blood was used as the perfusate. In an in vivo pig-to-primate heterotopic cardiac xenotransplantation model, in which porcine Xgs transplanted into untreated cynomolgus monkey recipients underwent HAR in 1 hr or less (n = 3), a single intravenous bolus of sCR1 (15 mg/kg) administered to the recipient immediately before Xg reperfusion markedly inhibited total and alternative pathway serum C activity and prolonged Xg survival to between 48 and 90 hr (n = 5). These studies confirm the important role of C activation in HAR of porcine cardiac Xgs by primates and indicate that sCR1 may be a useful agent for xenotransplantation.

An unusual case of mixed-dust exposure involving a "noncommercial" asbestos.

Our health center evaluated an individual for suspected pneumoconiosis, which had resulted from exposures in a foundry/metal reclamation facility. Appropriate consent forms were obtained for the procedures. Historically, individuals who work in foundries have been exposed to various types of dusts. The clinical findings in this case were consistent with silicosis with a suspicion of asbestos-induced changes as well. A sample from this individual, analyzed by electron microscopy, showed both classical and atypical ferruginous bodies. The uncoated fiber burden in this individual indicated an appreciable number of anthophyllite asbestos fibers. This finding, coupled with analysis of cores from ferruginous bodies and the presence of ferruginous bodies in areas of interstitial fibrosis, pathologically supported the diagnosis of asbestos-related disease. The unique factor associated with this case is that unlike in some settings in Finland where anthophyllite was mined and used commercially, this mineral fiber is not commonly found in commercially used asbestos products in the United States. Although the actual source of the asbestos exposure in this case is still being sought, it should be recognized that anthophyllite is a contaminant of many other minerals used in workplace environments, including foundries. The fiber burden indicates a unique type of exposure, differing from that usually construed as typical in occupational settings in the United States.

The effect of soluble complement receptor 1 (sCR1) and human thyroid antibodies on the course of experimental autoimmune thyroiditis in rats.

Experimental autoimmune thyroiditis (EAT), induced by immunisation of rats with thyroid extract and complete Freund's adjuvant, has been used as a model to study the effects of complement inhibition mediated by soluble complement receptor 1 (sCR1) administration during the initial phase of the disease. There was no effect of sCR1 on the severity of thyroiditis at day 28 after immunisation or on the levels of thyroid antibodies, whether sCR1 was given during the first or second week after immunisation. Human IgG containing high levels of thyroid peroxidase antibodies given to rats at the time of immunisation caused significant worsening of thyroiditis severity (P < 0.01 compared to animals receiving normal IgG) but sCR1 again had no effect in this variant of the EAT model. The results indicate that complement does not play a major role in the initial phase of tissue injury in EAT and complement inhibition does not impair the generation of an autoimmune response against the thyroid, although it remains possible that complement activation is important during the chronic phase of disease maintenance in human autoimmune thyroid disease.

Effect of soluble complement receptor type 1 on porphyrin-induced phototoxicity in guinea pigs.

The effect of recombinant human soluble complement receptor type 1 (sCR1) on the porphyrin-mediated phototoxic reaction was evaluated in guinea pigs. Phototoxicity was induced in the animals by intraperitoneal injection of hematoporphyrin derivative (HpD), followed by irradiation at a wavelength range of 320-450 nm. sCR1 administration decreased CH50 titers in a dose-dependent fashion, while it only moderately suppressed HpD/radiation-induced ear swelling at a high dose. These findings suggest that phototoxic dermatological changes in cutaneous porphyrias are not solely due to complement activation.

Pseudocoarctation of the aortic arch in a patient with Turner's syndrome.

Pseudocoarctation of the aortic arch is a rare congenital anomaly which resembles true coarctation by the presence of a narrowing in the descending thoracic aorta immediately distal to the origin of the left subclavian artery. However, it is differentiated from that entity by the absence of a significant hemodynamic obstruction and by the presence of aneurysmal dilatation proximal and distal to the area of narrowing. As a result, the commonly observed clinical findings of coarctation such as a prominent collateral circulation and decreased blood pressure in the lower extremities are not observed. "Kinking" or "buckling" are common adjectives used to describe the roentgenographic appearance of the aortic arch in this condition. The association of various congenital cardiac abnormalities with pseudocoarctation has been reported, and are similar to those commonly seen with true coarctation. We now present the first reported case of pseudocoarctation associated with a probable bicuspid aortic valve in a patient with Turner's syndrome.

Azoospermia and oligospermia among a large cohort of DBCP applicators in 12 countries.

Azoospermia and oligospermia have been well demonstrated among workers exposed to 1,2-dibromo-3-chloropropane (DBCP) in manufacturing and formulation of this pesticide. After DBCP was banned in the United States in the late 1970s, two American companies continued to export it to many less developed countries. In the early to mid-1990s, attorneys assembled a cohort of approximately 26,400 male plaintiffs who, as workers on banana and pineapple plantations in 12 of these countries, had been exposed to DBCP, primarily during its application. These attorneys, for the purpose of a lawsuit against the two American companies, developed from interrogatories a database on these men that included information about stated periods of occupational DBCP exposure. Seminal fluid analysis results were also entered into the database. Analyzing information in this database, the authors found that, after a median exposure to DBCP of three years, 64.3% of these men overall, and 90.1% of men studied from the Philippines, had azoospermia or oligospermia. The mean number of children reported by the men was 2.5 overall. The percentage of men with no children was 28.5% overall. This report represents the largest cohort of DBCP-exposed workers in which adverse reproductive health effects have been described, and the first report of the adverse effects on the reproductive health of workers exposed to DBCP primarily through its application in a cohort of this size. This serious and extensive occurrence of adverse reproductive health effects due to the export of a hazardous pesticide before and after its ban in the United States illustrates a number of needs for monitoring, research, education, and policy development.

An individual with a majority of ferruginous bodies formed on chrysotile cores.

A 59-year-old man was exposed to chrysotile asbestos while rebuilding clutches. Analytical electron microscopy showed a chrysotile core in 72% of the ferruginous bodies from lung tissue. Long, uncoated chrysotile fibers were also present. Sufficient exposure to long chrysotile in jobs such as this appears to allow the majority of ferruginous bodies to be formed on chrysotile, an exception to the rule that most ferruginous bodies form on amphibole cores.

Respiratory effects of outdoor air pollution.

Outdoor air pollution adversely affects human health and the quality of the environment. However, epidemiologic studies of these effects are difficult to control because of confounding variables such as age and cigarette smoking and the difficulty in estimating doses of pollutants. Drs Griffith and Levin discuss the relationship between major types of pollutants and increased morbidity and mortality from respiratory disease.

Symposium on environmental medicine: into the next millennium.

Physicians and other health care providers are witnessing a growth in the number of patients who present with concerns related to environmental exposures. Environmental medicine emphasizes evaluation and prevention of exposure-related illness. Both patients and their communities view physicians as credible sources of information about environmental health matters; however, the knowledge and training that physicians have in this area are generally limited. Recognizing this shortcoming, the Institute of Medicine (IOM) has recommended that competency-based training be integrated into all levels of medical education for physicians. This symposium issue includes articles written by Texas physicians from various forms of practice, and complements the IOM learning approach by serving as a resource of information. The goal is to increase knowledge and awareness of environmental issues among physicians who play a special stewardship role for our planet and the health of its human inhabitants.

The Texarkana mercury incident.

In November 1997, 2 teenagers allegedly removed a large amount of metallic mercury from an abandoned sign plant and distributed the material among friends. One teenager developed symptoms and admitted playing with mercury to his physician. His blood mercury was elevated. In February 1998, faculty from the University of Texas Health Center at Tyler conducted an investigation that included in-depth evaluations on 10 patients with urine mercury concentrations up to 100 micrograms/L. Exposure pathways and timelines were reconstructed from records assembled by the Arkansas State Health Department epidemiologist. Mercury contamination was found among teenagers, children, and adults who came in contact with the metal. Biomarkers of exposure documented reduction in mercury concentrations after these persons were removed from their homes and sources of mercury. Neurobehavioral assessment, including assessment of tremor, failed to establish a relationship between mercury exposure and performance.