Additive Manufacturing for Occupational Hygiene: A Comprehensive Review of Processes, Emissions, & Exposures.

This comprehensive review introduces occupational (industrial) hygienists and toxicologists to the seven basic additive manufacturing (AM) process categories. Forty-six articles were identified that reported real-world measurements for all AM processes, except sheet lamination. Particles released from powder bed fusion (PBF), material jetting (MJ), material extrusion (ME), and directed energy deposition (DED) processes exhibited nanoscale to submicron scale; real-time particle number (mobility sizers, condensation nuclei counters, miniDiSC, electrical diffusion batteries) and surface area monitors (diffusion chargers) were generally sufficient for these processes. Binder jetting (BJ) machines released particles up to 8.5 µm; optical particle sizers (number) and laser scattering photometers (mass) were sufficient for this process. PBF and DED processes (powdered metallic feedstocks) released particles that contained respiratory irritants (chromium, molybdenum), central nervous system toxicants (manganese), and carcinogens (nickel). All process categories, except those that use metallic feedstocks, released organic gases, including (but not limited to), respiratory irritants (toluene, xylenes), asthmagens (methyl methacrylate, styrene), and carcinogens (benzene, formaldehyde, acetaldehyde). Real-time photoionization detectors for total volatile organics provided useful information for processes that utilize polymer feedstock materials. More research is needed to understand 1) facility-, machine-, and feedstock-related factors that influence emissions and exposures, 2) dermal exposure and biological burden, and 3) task-based exposures. Harmonized emissions monitoring and exposure assessment approaches are needed to facilitate inter-comparison of study results. Improved understanding of AM process emissions and exposures is needed for hygienists to ensure appropriate health and safety conditions for workers and for toxicologists to design experimental protocols that accurately mimic real-world exposure conditions.ABBREVIATIONS ABS : acrylonitrile butadiene styrene; ACGIH® TLV® : American Conference of Governmental Industrial Hygienists Threshold Limit Value; ACH : air change per hour; AM : additive manufacturing; ASA : acrylonitrile styrene acrylate; AVP : acetone vapor polishing; BJ : binder jetting; CAM-LEM : computer-aided manufacturing of laminated engineering materials; CNF : carbon nanofiber; CNT : carbon nanotube; CP : co-polyester; CNC : condensation nuclei counter; CVP : chloroform vapor polishing; DED : directed energy deposition; DLP : digital light processing; EBM : electron beam melting; EELS : electron energy loss spectrometry; EDB : electrical diffusion batteries; EDX : energy dispersive x-ray analyzer; ER : emission rate; FDM™ : fused deposition modeling; FFF : fused filament fabrication; IAQ : indoor air quality; LSP : laser scattering photometer; LCD : liquid crystal display; LDSA : lung deposited particle surface area; LOD : limit of detection; LOM : laminated object manufacturing; LOQ : limit of quantitation; MCE : mixed cellulose ester filter; ME : material extrusion; MJ : material jetting; OEL : occupational exposure limit; OPS : optical particle sizer; PBF : powder bed fusion; PBZ : personal breathing zone; PC : polycarbonate; PEEK : poly ether ether ketone; PET : polyethylene terephthalate; PETG : Polyethylene terephthalate glycol; PID : photoionization detector; PLA : polylactic acid; PM1 : particulate matter with aerodynamic diameter less than 1 µm; PM2.5 : particulate matter with aerodynamic diameter less than 2.5 µm; PM10 : particulate matter with aerodynamic diameter less than 10 µm; PSL : plastic sheet lamination; PVA : polyvinyl alcohol; REL : recommended exposure limit; SDL : selective deposition lamination; SDS : safety data sheet; SEM : scanning electron microscopy; SL : sheet lamination; SLA : stereolithography; SLM : selective laser melting; SMPS : scanning mobility particle sizer; SVOC : semi-volatile organic compound; TEM : transmission electron microscopy; TGA : thermal gravimetric analysis; TPU : thermo polyurethane; UAM : ultrasonic additive manufacturing; UC : ultrasonic consolidation; TVOC : total volatile organic compounds; TWA : time-weighted average; VOC : volatile organic compound; VP : vat photopolymerization.

Reflections on the OECD guidelines for in vitro skin absorption studies.

At the 8th conference of Occupational and Environmental Exposure of the Skin to Chemicals (OEESC) (16-18 September 2019) in Dublin, Ireland, several researchers performing skin permeation assays convened to discuss in vitro skin permeability experiments. We, along with other colleagues, all of us hands-on skin permeation researchers, present here the results from our discussions on the available OECD guidelines. The discussions were especially focused on three OECD skin absorption documents, including a recent revision of one: i) OECD Guidance Document 28 (GD28) for the conduct of skin absorption studies (OECD, 2004), ii) Test Guideline 428 (TGD428) for measuring skin absorption of chemical in vitro (OECD, 2004), and iii) OECD Guidance Notes 156 (GN156) on dermal absorption issued in 2011 (OECD, 2011). GN156 (OECD, 2019) is currently under review but not finalized. A mutual concern was that these guidance documents do not comprehensively address methodological issues or the performance of the test, which might be partially due to the years needed to finalize and update OECD documents with new skin research evidence. Here, we summarize the numerous factors that can influence skin permeation and its measurement, and where guidance on several of these are omitted and often not discussed in published articles. We propose several improvements of these guidelines, which would contribute in harmonizing future in vitro skin permeation experiments.

Assessing a portable, real-time display handheld meter with UV-A and UV-B sensors for potential application in personal sun exposure studies.

BACKGROUND: Observing accurate real-time measurements of solar ultraviolet radiation (UVR) levels is important since personal excess sun exposure is associated with skin cancers. Handheld measurement devices may be helpful but their accuracy is unknown. We compare a portable, science-grade solar UVR monitoring device against two fixed, science-grade solar UVR instruments. METHODS: Instruments were (1) a fixed Solar Light 501 UV-B biometer to measure UV-B; (2) a fixed Kipp and Zonen radiometer used to measure UV-A and UV-B; and (3) Goldilux ultraviolet probes which are commercially available portable devices. Two different probes were used, one measured UV-A and the other UV-B radiation. The Goldilux probes were levelled and secured next to the UV-B biometer. Between 10:00 and 14:40 UTC+2, the UV-B biometer was set to record at 10-minute intervals and measurements by the Goldilux probes were manually taken simultaneously. Results were compared for all data and by solar zenith angle (SZA) ranges. RESULTS: The Goldilux UV-B probe measured UV-B relatively well in its diurnal pattern, however, its readings were ~77% higher than those made by the UV-B biometer. While UV-A measurements from the Goldilux UV-A probe and those from the radiometer were in relatively good agreement in pattern, the radiometer read ~47% higher than the Goldilux UV-A probe. UV-B data from Goldilux UV-B probe had a moderately strong correlation with UV-B biometer data for small SZAs; conversely, for UV-A, the Goldilux UV-A probe had a strong correlation with the UV-A radiometer data for large SZAs. CONCLUSION: Handheld devices may be useful to provide real-time readings of solar UVR patterns, however, to achieve synchronicity in the magnitude of readings to those made by science-grade fixed instruments, devices may need to be used during certain times of the day and in clear-sky conditions which may not be practical in personal exposure studies.

In multiple situational light settings, visual observation for skin colour assessment is comparable with colorimeter measurement.

BACKGROUND: Finding inexpensive and reliable techniques for assessing skin colour is important, given that it is related to several adverse human health outcomes. Visual observation is considered a subjective approach assessment and, even when made by trained assessor, concern has been raised about the need for controlled lighting in the study venue. The aim of this study is to determine whether visual skin colour assessments correlate with objective skin colour measurements in study venues with different lighting types and configurations. METHODS: Two trained investigators, with confirmed visual acuity, visually classified the inner, upper arm skin colour of 556 adults using Munsell(®) colour classifications converted to Individual Typology Angle (°ITA) values based on published data. Skin colour at the same anatomic site was also measured using a colorimeter. Each participant was assessed in one of 10 different buildings, each with a different study day. Munsell(®) -derived °ITA values were compared to colorimeter °ITA values for the full sample and by building/day. RESULTS: We found a strong positive, monotonic correlation between Munsell(®) derived °ITA values and colorimeter °ITA values for all participants (Spearman ρ = 0.8585, P < 0.001). Similar relationships were found when Munsell(®) and colorimeter °ITA values were compared for participants assessed in the same building for all 10 buildings (Spearman ρ values ranged from 0.797 to 0.934, all correlations were statistically significant at P < 0.001). CONCLUSION: It is possible to visually assess individual skin colour in multiple situational lighting settings and retrieve results that are comparable with objective measurements of skin colour. This was true for individuals of varying population groups and skin pigmentation.

Insights Into Emissions and Exposures From Use of Industrial-Scale Additive Manufacturing Machines.

BACKGROUND: Emerging reports suggest the potential for adverse health effects from exposure to emissions from some additive manufacturing (AM) processes. There is a paucity of real-world data on emissions from AM machines in industrial workplaces and personal exposures among AM operators. METHODS: Airborne particle and organic chemical emissions and personal exposures were characterized using real-time and time-integrated sampling techniques in four manufacturing facilities using industrial-scale material extrusion and material jetting AM processes. RESULTS: Using a condensation nuclei counter, number-based particle emission rates (ERs) (number/min) from material extrusion AM machines ranged from 4.1 × 1010 (Ultem filament) to 2.2 × 1011 [acrylonitrile butadiene styrene and polycarbonate filaments). For these same machines, total volatile organic compound ERs (µg/min) ranged from 1.9 × 104 (acrylonitrile butadiene styrene and polycarbonate) to 9.4 × 104 (Ultem). For the material jetting machines, the number-based particle ER was higher when the lid was open (2.3 × 1010 number/min) than when the lid was closed (1.5-5.5 × 109 number/min); total volatile organic compound ERs were similar regardless of the lid position. Low levels of acetone, benzene, toluene, and m,p-xylene were common to both AM processes. Carbonyl compounds were detected; however, none were specifically attributed to the AM processes. Personal exposures to metals (aluminum and iron) and eight volatile organic compounds were all below National Institute for Occupational Safety and Health (NIOSH)-recommended exposure levels. CONCLUSION: Industrial-scale AM machines using thermoplastics and resins released particles and organic vapors into workplace air. More research is needed to understand factors influencing real-world industrial-scale AM process emissions and exposures.

Evaluation of emissions and exposures at workplaces using desktop 3-dimensional printer.

There is a paucity of data on additive manufacturing process emissions and personal exposures in real-world workplaces. Hence, we evaluated atmospheres in four workplaces utilizing desktop "3-dimensional" (3-d) printers [fused filament fabrication (FFF) and sheer] for production, prototyping, or research. Airborne particle diameter and number concentration and total volatile organic compound concentrations were measured using real-time instruments. Airborne particles and volatile organic compounds were collected using time-integrated sampling techniques for off-line analysis. Personal exposures for metals and volatile organic compounds were measured in the breathing zone of operators. All 3-d printers that were monitored released ultrafine and fine particles and organic vapors into workplace air. Particle number-based emission rates (#/min) ranged from 9.4 × 109 to 4.4 × 1011 (n = 9samples) for FFF3-d printers and from 1.9 to 3.8 × 109 (n = 2 samples) for a sheer 3-d printer. The large variability in emission rate values reflected variability from the printers as well as differences in printer design, operating conditions, and feedstock materials among printers. A custom-built ventilated enclosure evaluated at one facility was capable of reducing particle number and total organic chemical concentrations by 99.7% and 53.2%, respectively. Carbonyl compounds were detected in room air; however, none were specifically attributed to the 3-d printing process. Personal exposure to metals (aluminum, iron) and 12 different organic chemicals were all below applicable NIOSH Recommended Exposure Limit values, but results are not reflective of all possible exposure scenarios. More research is needed to understand 3-d printer emissions, exposures, and efficacy of engineering controls in occupational settings.

Southern African scorpion toxins: an overview.

In southern Africa there are 130 species of scorpions and only a few species' venom have been investigated to date. This review gives an overview of the research done on the venom of southern African scorpions and the toxins and peptides identified up to date. It also aims to highlight the enormous potential that lies in this field of research. Southern African scorpion toxins include four long-chain Na(+)-channel toxins, four short-chain alpha-K(+)-channel toxins (alpha-KTx), three Ca(2+)-channel toxins and also an insect-selective peptide active on K(+) and Cl(-) channels. Three antimicrobial peptides have also been isolated and characterized. All of these peptides are diverse not only in function and target but also in the species they are isolated from.

DNA damage and repair detected by the comet assay in lymphocytes of african petrol attendants: a pilot study.

Petrol attendants are exposed to petrol volatile organic compounds (VOCs) which may have genotoxic and carcinogenic effects. The single-cell gel electrophoresis assay (comet assay) is a method highly sensitive to DNA damage induced by environmental and occupational exposure to carcinogenic and mutagenic agents. The aim of this study was to evaluate the level of exposure of petrol attendants to petrol VOCs and also to determine their effect on DNA damage and repair in lymphocytes of African petrol attendants. The exposed group consisted of 20 subjects, randomly selected from three petrol stations. A control group of 20 unexposed subjects was also chosen and matched for age and smoking habits with the exposed group. Sorbent tubes were used to assess personal exposure of petrol attendants. The comet assay was used to investigate the basal DNA damage and repair capacity in isolated lymphocytes of petrol attendants and unexposed subjects. Blood samples were taken from the petrol attendants at the end of their 8-h working shift and also from the unexposed subjects. The petrol attendants were found to be exposed to levels of petrol VOCs lower than the South African occupational exposure limit for constituent chemicals. A significant relationship was found between the volume of petrol sold during the shift and the average concentrations of benzene, toluene and the total VOCs measured. However, relative humidity had a negative correlation with the average concentrations of benzene, toluene, xylene and the total VOCs. Significantly higher basal DNA damage was observed with the exposed group compared to the unexposed group. The period of exposure influenced the level of DNA damage and the calculated repair capacity. Smoking and age had a significant influence on the level of DNA damage. DNA repair capacity was delayed in smokers of both exposed and unexposed group.

In vitro permeation of platinum through African and Caucasian skin.

The majority of the South African workforce are Africans, therefore potential racial differences should be considered in risk and exposure assessments in the workplace. Literature suggests African skin to be a superior barrier against permeation and irritants. Previous in vitro studies on metals only included skin from Caucasian donors, whereas this study compared the permeation of platinum through African and Caucasian skin. A donor solution of 0.3 mg/ml of potassium tetrachloroplatinate (K2PtCl4) dissolved in synthetic sweat was applied to the vertical Franz diffusion cells with full thickness abdominal skin. Skin from three female African and three female Caucasian donors were included (n=21). The receptor solution was removed at various intervals during the 24 h experiment, and analysed with high resolution inductively coupled plasma-mass spectrometry (ICP-MS). Skin was digested and analysed by inductively coupled plasma-optical emission spectrometry (ICP-OES). Significantly higher permeation of platinum through intact African skin (p=0.044), as well as a significantly higher mass of platinum retention in African skin in comparison with Caucasian skin (p=0.002) occurred. Significant inter-donor variation was found in both racial groups (p<0.02). Results indicate that African workers have increased risk of dermal permeation and therefore possible sensitisation caused by dermal exposure to platinum salts. These results are contradictory to limited literature suggesting a superior barrier in African skin and further investigation is necessary to explain the higher permeation through African skin.

Different organisms associated with heartwater as shown by analysis of 16S ribosomal RNA gene sequences.

Cowdria ruminantium is a rickettsial parasite which causes heartwater, a economically important disease of domestic and wild ruminants in tropical and subtropical Africa and parts of the Caribbean. Because existing diagnostic methods are unreliable, we investigated the small-subunit ribosomal RNA (srRNA) gene from heartwater-infected material to characterise the organisms present and to develop specific oligonucleotide probes for polymerase chain reaction (PCR) based diagnosis. DNA was obtained from ticks and ruminants from heartwater-free and heartwater-endemic areas from Cowdria in tissue culture. PCR was carried out using primers designed to amplify only rickettsial srRNA genes, the target region being the highly variable V1 loop. Amplicons were cloned and sequenced; 51% were C. ruminantium sequences corresponding to four genotypes, two of which were identical to previously reported C. ruminantium sequences while the other two were new. The four different Cowdria genotypes can be correlated with different phenotypes. Tissue-culture samples yielded only Cowdria genotype sequences, but an extraordinary heterogeneity of 16S sequences was obtained from field samples. In addition to Cowdria genotypes we found sequences from previously unknown Ehrlichia spp., sequences showing homology to other Rickettsiales and a variety of Pseudomonadaceae. One Ehrlichia sequence was phylogenetically closely related to Ehrlichia platys (Group II Ehrlichia) and one to Ehrlichia canis (Group III Ehrlichia). This latter sequence was from an isolate (Germishuys) made from a naturally infected sheep which, from brain smear examination and pathology, appeared to be suffering from heartwater; nevertheless no Cowdria genotype sequences were found in this isolate. In addition no Cowdria sequences were obtained from uninfected ticks. Complete 16S rRNA gene sequences were determined for two C. ruminantium genotypes and for two previously uncharacterised heartwater-associated Ehrlichia spp. Sequenced difference within the V1 loop were sufficient for the derivation of four Cowdria genotype-specific oligonucleotide probes. Four further probes were designed; one for the detection of any Cowdria genotype, one for the detection of any Group II Ehrlichia sp., one for any Group III Ehrlichia sp. and one for all Pseudomonadaceae. All the probes were specific except that for the Cowdria (Ball 3) genotype. The high prevalence (96%) in field samples of pseudomonad-like 16S sequences was the result of environmental contamination. The probes were used to screen DNA from goats in an area free of both Amblyomma ticks and clinical heartwater. A substantial proportion (42%) gave positive reactions for the apparently apathogenic Cowdria (Omatjenne), indicating that this genotype is relatively common.

A method for determining the Cowdria ruminantium infection rate of Amblyomma hebraeum: effects in mice injected with tick homogenates.

Amblyomma hebraeum ticks, collected in the field and individually homogenized, were injected into mice. Thirteen out of 240 ticks were shown to be infected with the heartwater agent. Antibodies against Cowdria ruminantium were detected in the sera of the mice by means of the indirect fluorescent antibody test. Giemsa-stained smears, prepared from the haemocytes of the ticks, revealed morphologically different forms of the heartwater agent. A strain of C. ruminantium, designated the Welgevonden strain, was isolated in mice from one of the infected ticks and passaged in mice for 8 generations. When inoculated intravenously, it was highly infective to mice, sheep and cattle. The murinotropism of the Welgevonden strain is compared with that of other strains previously described.

The natural resistance of cattle to artificial infection with Cowdria ruminantium: the role played by conglutinin.

The conglutinin titres of year-old Bonsmara-cross cattle infected with Cowdria ruminantium were inversely proportional to the severity of the reactions elicited by the infection. There was no correlation, however, between conglutinin levels of 8-month-old calves of the same breed, sex and origin and their susceptibility to heartwater. The role possibly played by conglutinin in the non-specific resistance of cattle to heartwater and in the epidemiology of the disease is discussed.

An in vitro test to demonstrate the inhibitory effect of homologous immune serum on the infectivity of Cowdria ruminantium.

Homologous immune sheep serum inhibits the infectivity of mice for the Kümm stock of Cowdria ruminantium in an in vitro test. The presence of complement and an optimal ratio between the reagents are of cardinal importance. With the exception of the Ball 3 stock, there is no cross-inhibition between the immune sera of sheep infected with several other heterologous stocks and the Kümm stock. The role played by the humoral component of the immune response in naturally acquired heartwater is unknown, but probably small compared to the cellular component.

Increased pathogenicity of an Ehrlichia-like agent after passage through Amblyomma hebraeum: a preliminary report.

After being passaged through 3 generations of Amblyomma hebraeum, an Ehrlichia-like agent isolated from an adult Hyalomma truncatum female became more pathogenic and elicited a disease in sheep indistinguishable from heartwater. Cross-immunity between this agent and several stocks of Cowdria ruminantium and high levels of antibody elicited by the agent against 2 stocks of C. ruminantium in the indirect fluorescent antibody test, confirmed its close relationship to Cowdria.

The influence of dithiosemicarbazone on the immunity of sheep to heartwater.

Four out of 9 sheep, immune to heartwater and subsequently treated with gloxazone, showed a febrile reaction when they were challenged 6 months later, but the same number of untreated controls also developed this reaction. In a second group of treated immune animals challenged after 9 months, 8 out of 8 showed a febrile reaction, whereas only 3 out of 8 controls reacted. Furthermore, the blood of one of the 4 sheep that reacted to challenge at 6 months was infective to a susceptible sheep inoculated with it, whereas that of 4 out of the 8 challenged at 9 months was infective.

Colostrum-derived antibodies to Cowdria ruminantium in the serum of calves and lambs.

Antibodies to Cowdria ruminantium were detected in the serum of calves born from artificially immunized heifers, by means of the indirect fluorescent antibody test, only for as long as 4 weeks after birth. Lambs born from artificially immunized ewes, however, were still serologically positive at 8-12 weeks of age. Much higher antibody titres were recorded in the sera of ewes and their lambs than in that of heifers and their calves.

Electron microscopy of Cowdria-infected macrophages suggests that in the absence of binary fission a mosaic of organisms develops from an amorphous electron dense matrix.

Electron microscopy of mouse peritoneal macrophages infected with the Kümm stock of Cowdria ruminantium suggests that in the final stage of intracellular growth, a mosaic of organisms develops from an amorphous matrix of varying electron density by a process in which double unit membranes portion off the Cowdria particles. This stage is preceded by inclusions consisting of a network of aggregated electron dense granules and these in turn by homogeneous dense bodies. The study failed to show how these dense bodies develop from internalized Cowdria particles introduced in the infective inoculum. The replication of the heartwater agent in macrophages differs from that in vascular endothelial cells in two important respects. First, at no stage during the course of development in macrophages is binary fission in evidence and second, in the absence of a limiting membrane the inclusions and colonies of organisms throughout the cycle of development in macrophages are in intimate contact with the host cell cytoplasm.

Susceptibility to heartwater of calves born to non-immune cows.

The resistance to artificial infection with Cowdria ruminantium of calves born to cows fully susceptible to heartwater is no different from that of calves bred in heartwater endemic areas where the tick challenge is negligible to considerable. The sub-inoculation into mice of blood collected 14-26 days after infection proved the presence of the heartwater agent in the blood of 8 out of 10 calves with no other clinical signs than mild to moderate fever. The combined use of a mouse model and the indirect fluorescent antibody test revealed considerable variation in the degrees to which calves become infected and react to artificial infection.

Heartwater in Angora goats. II. A pathological study of artificially infected, treated and untreated goats.

Pathological lesions in untreated Angora goats infected with the Ball3 strain of Cowdria ruminantium corresponded with those previously reported. A severe nephrosis was the most prominent pathological lesion in the animals treated after the 1st day of the febrile reaction. Renal ischaemia appears to be central to the pathogenesis of the kidney lesions.