A PRIL 2014, N O 6
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Microbial Contamination of the Russian Manned Spacecraft A total of 234 species of bacteria and microscopic fungi were identified in the Mir environment. The bacterial flora consisted of 108 species, whereas the fungal flora included 126 species.Opportunistic pathogens of opportunistic infections were noted among bacteria, and pathogenic saprophytes causing mycoses and mycointoxications were observed among fungi. The greatest diversity (64 species) was demonstrated by the group of technophylic fungi that destroy polymers and corrode metals. Out of 1150 samples gathered from Mir surfaces and air to study bacterial flora, the microorganisms were found in 949 samples (82.5% of the total number of samples). Table 1 contains data on occurrence of different bacterial genera in the space station environment.
Table 2 presents data on the occurrence of various fungal genera on the orbital station. Penicillium, Aspergillus, and Cladosporium dominated by occurrence in samples of the interior and equipment surfaces as well as in air samples. On the whole, they reached 76.8 - 75.8%, 39.4 - 76.6% and 27.0 - 24.2%, respectively.
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Contamination of Air Bacterial flora of air has been composed of 19 genera and 46 species. At all times, the special structure of bacteria consisted primarily of commensals of human integument, i.e. staphylococci, micrococci, and coryneform bacteria. Streptococci and enterococci were occasional. In addition, air samples were periodically analyzed for residents of natural reservoirs, first of all for spore-forming bacteria g.Bacillus and gram-negative non-fermenting bacteria. The highest occurrence was characteristic of genes Staphylococcus(53.2%), Bacillus (34.0%), and Corynebacterium (16.0%). A significant number of microbial species in air were opportunistic pathogens including Staphylococcus aureus, S. capitis, S. haemoliticus, Flavobacterium meningosepticum,Escherichia coli, Serratia marcescens, Streptococcussp., Bacillus cereus. Micromycets from air displayed large diversity. Fungi were presented by 62 species and 13 genes. The population of aeroplankton was dominated by Pencillins; Apergills were also of weight.Micromycets of Penicillium, Aspergillus and Cladosporium were the most numerous findings in air. In Mir’s operational lifetime, bacterial contamination of its air remained relatively stable and did not exceed the ISOPTWPO Today Human Space Flight c International Space Agency(ISA)
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limit (500 CFU/m3 ) in 95% of the air samples. The relatively high bacterial contamination of air, which in several instances reached 3.53 103 CFU in m3 , was observed at the site of exercise machines in the basal module, in Kvant - 2 and Kristall. The highest occurrence on structural materials was characteristic of Corynebacterium sp. and Staphylococcus epidermidis (33.2% and 30.0%, respectively). Contamination of Surfaces The interior site samples contained potential agents of opportunistic infections of the Enterobacteriaceae family (Enterobacter, Escherichia, Proteus, Serratia, and other genera), and bacilli Staphylococcus aureus. The latter were isolated in 2.3% of the total number of samples. Bacillus sp. also exhibited high occurrence; for instance, Bacillus subtilis were found in 5.2%, and Bacillus sphaericus in 5.0% of the total number of samples. Altogether, 116 fungal species of 25 genera were isolated and identified in the period under review. The microflora of the interior and equipment was dominated by representatives of Penicillium, Aspergillus, and Cladosporium sp., among which the most frequent were Penicillium expansum (28.9% of the samples),Penicillium chrysogenum and Cladosporium cladosporioides(16.8% and 17.0% of the samples, respectively),Aspergillus sp. of group A. versicolor (15.5% of the samples), Aspergillus versicolor (10.0% of the samples),and Aspergillus niger (7.9% of the samples). The majority of the fungal species on structural materials are well-known potential biodegradors of polymers and, therefore, can be considered agents that may produce biointerference, damage to structural materials, malfunctioning and failure of various space systems and equipment. Besides, a number of isolated micromycets are known to affect human organism as etiological agents of infections and allergies . Microbiological analysis of debris from Space Transportation System (STS) - 55 Spacelab D - 2 Filter debris from the Spacelab module D - 2 of STS - 55 was analyzed for microbial contamination. Debris from cabin and avionics filters was collected by Kennedy Space Center personnel on May 8, 1993, 2 days postflight. Debris weights were similar to those of previous Spacelab missions. Approximately 5.1E + 5 colony forming units per gram of debris were enumerated from the cabin and avionics filter debris, respectively. these numbers were similar in previous missions for which the entire contents were analyzed without sorting of the material. Bacterial diversity was small compared to previous missions, with no gram negative bacteria isolated. Only one bacterial species, Corynebacterium pseudodiphtheriticum, was not isolated previously by the laboratory from Spacelab debris. This organism is a normal inhabitant of the pharynx. A table listing all species of bacteria isolated by the laboratory from previous Spacelab air filters debris collection is provided. Reference: 1.Review of the Knowledge of Microbial Contamination of the Russian Manned Spacecraft,N.D. Novikova 2.Microbiological analysis of debris from Space Transportation System (STS)-55 Spacelab D-2,JAXA
Changes in the Fungal Autoflora of Apollo Astronauts
A summary of the genera of yeasts and filamentous fungi isolated from crew specimens throughout the Apollo 14 and 15 missions is presented in Table 3. In all, 112 species, belonging to 57 genera, were identified from these specimens.Fifty-three percent of these genera were recovered during no more than one period per mission and are marked as rarely isolated genera in Table 3.Of the remaining 27 genera, 13 were isolated frequently (during four or more sample periods). However, in the case of the genera Aspergillus and Penicillium, there were many different representatives within the genus, so that most species of these genera were actually rarely isolated.
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Only those species which were isolated frequently (recovered at least once during four or more sample periods) are considered in detail. A complete listing of these 10 species is presented in Table 4.
These include one species each from the genera Aureobasidium, Pityrosporum, and Scolescobasidium, with all others belonging to the genera Candida and Cladosporium. Candida albicans was the most ubiquitous microorganism, being recovered during each of the nine sample periods a total of 42 times. Other species of Candida were also frequently isolated, with the four species listed in Table 4 accounting for 60% of all the isolates outlined on this table. Similarly, three species of Cladosporium account for 26% of all isolates from the frequently occurring microorganisms presented in Table 4. Scolescobasidium verruculosm was the least frequently isolated species of this group, being recovered from the Apollo 14 crew only. A tabulation of the mycological recovery of all crewmembers for both missions is presented in Table 5. Because the frequently isolated species recovered throughout the Apollo 14 and 15 missions are restricted to five genera, only these genera were evaluated for specific variations throughout the two missions. All of the members of these five genera, isolated during the Apollo 14 and 15 missions, are presented in Tables 6 and 7 respectively.
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Reference:GERALD R. TAYLOR, MARY R. HENNEY, AND WALTER L. ELLIS: Changes in the Fungal Autoflora of Apollo Astronauts
Autoflora in the Upper Respiratory Tract of Apollo Astronauts
Recovery of anaerobic bacteria from nasal passages of Apollo 13 and 14 crew members An average of 1.2 species of anaerobic bacteria with a mean quantitation of 2,520 cells/cm2 of surface area was recovered from the nasal passages of each astronaut at each sample time before flight. The average recovery was 1.0 species, with a mean quantitation of 41,700 cells/cm2 the day of return from space flight. Although representatives of the genera Peptococcus and Peptostreptococcus were infrequently recovered from the nasal passages, only the genus Propionibacterium was isolated frequently enough to be considered a component of the nasal autoflora. Accordingly, only that genus is evaluated here. The preflight recovery pattern of this genus, outlined in Table 2, demonstrates no statistically significant (P < 0.05) difference between the five crew members, indicating that propionibacteria regularly inhabited each crew member at about the same quantitative level. Such regularity provided an opportunity to evaluate the effect of short-duration (Apollo 13, 143 h; Apollo 14, 217 h) spaceflight on the stable portion of the nasal autoflora. The data shown in Table 2 indicate that the recovery of viable cells, belonging to the genus Propionibacterium, from crew members immediately upon their return from spaceflight was not significantly (P < 0.05) different than for any preflight sample collection period, except in the case of astronaut 4. This value was higher than the values obtained from the other astronauts postflight and was also higher than the preflight values for this crew member. Because this postflight elevation was not duplicated by the other crew members, it canno be considered a typical response to space flight.
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Recovery of aerobic bacteria from nasal passages of Apollo 13 and 14 crew members The qualitative and quantitative recoveries of aerobic bacteria from the nasal passages of the five subject crew members at various times before and after space flight are shown in Table 3. When compared to preflight recovery, there is a marked decrease in the number of times the bacilli, corynebacteria, gram-negative rods, and streptococci were isolated immediately after flight. Simultaneous with this decrease,the incidence of the potentially pathogenic Staphylococcus aureus increased from a maximum of two isolates during any preflight period to a maximum of five postflight, being transferred to all of the crew members. Quantitatively, the total nasal population increased significantly (P < 0.05) postflight (Table 3), due to the accession of S. aureus mentioned above as well as an increase in quantitation of corynebacteria and Micrococcaceae.
Recovery of anaerobic bacteria from the oral cavity of crew members from Apollo 13 through 16 The quantitative and qualitative recovery of anaerobic bacterial genera from the oral cavity of the 11 subject astronauts is presented in Table 4 in decreasing frequency of isolation. Analysis of the generic recovery frequency data shows no major difference between the occurrence of any particular genus before and after flight. Similarly, quantitative analysis of the mean number of viable cells present belonging to any particular genus indicates that, in general, the postflight load was within the preflight range.
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Recovery of aerobic bacteria from the oral cavity of crew members from Apollo 13 through 16 crew members A summary of the recovery of aerobic bacteria from the gargle of 11 astronauts is outlined in Table 5. The family Micrococcaceae and the genera Streptococcus, Neisseria, and Haemophilus were generally recovered from each of the 44 gargle samples obtained. Only with the genus Haemophilus was the postflight recovery found to be significantly different from the preflight base line (P < 0.05). Members of the Enterobacteriaceae and other gram-negative rods were infrequently observed, and then only in relatively low numbers.
Reference:J. GLEE DECELLE AND GERALD R. TAYLOR,Autoflora in the Upper Respiratory Tract of Apollo Astronauts
Skylab Environmental and Crew Microbiology Studies Changes in the Habitability of the Skylab Environment Microbial Content of In-flight Skylab Air The concentration of bacteria recovered from air samples obtained 2 days before return from each Skylab visit are displayed in figure 1. Low levels of in-flight bacterial contamination were observed on the first two missions, whereas the recovery from Skylab 4 was considerably higher. These higher counts were due entirely to an influx of Serratia marcescens, a micro-organism which has been shown to produce various infections in man . Whereas this species was not recovered from any preflight crew sample analysis, it was recovered from multiple from all three Skylab 4 astronauts immediately upon recovery. Further, this species persisted in the nasal cavity of the Pilot throughout the postflight quarantine period. Subsequent investigation demonstrated several potential sources of this micro-organism in the Skylab environment.
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Bacterial Recovery from Sample Sites within the Skylab Orbital Workshop The total concentrations of viable bacterial cells recovered from the Skylab spacecraft surface sites at various sampling periods are presented in figure 2.
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These in-flight samples were collected to evaluate the level of microbial contamination occurring in the Orbital Workshop. The results of analyses of samples collected prior to launch are typical of a clean(although obviously not sterile) environment. The reduction of aerobic bacteria recovered from the Skylab 2 in-flight samples is probably a reflection of the thermal problems experienced in the Orbital Workshop after launch. Although there was a simultaneous tenfold increase in the presence of anaerobic bacteria, the Skylab 2 crew apparently entered a very clean environment, which remained relatively clean during the mission.The recovery of both aerobic and anaerobic bacteria from the Skylab 3 mission increased another 1 to 2 loglo units, with no apparent reason except for increased length of habitation by the crewmembers. During the 84 - day Skylab 4 mission the total concentration of aerobic bacteria remained nearly constant although anaerobe recovery decreased significantly. This drop was due to the loss of Propionibacterium acnes which contributed strongly to the anaerobe population of the other two Skylab missions. This loss of P. aches reflects a similar loss of anaerobic bacteria from the skin surfaces of the astronauts these data will be presented later in this paper). Therefore, this decrease in anaerobic bacterial contamination of the Skylab, was shown to directly reflect a decrease in these same microbes in the contaminating reservoir, the skin of the astronauts. The recovery of aerobic bacteria from 15 sites within the Apollo Command Modules, sampled immediately before and after each mission to the Skylab, are summarized in figure 3. Whereas there was some variation in the contamination level of the different Command Modules, there were no major differences between preflight and postflight values for a particular Command Module.
Fungal Recovery from Sample Sites within the Skylab Orbital Workshop Figure 4 shows the number of fungal isolations from the Skylab vehicle before launch and during each mission. These numbers were low until the Skylab 4 mission. Although overall humidity was low on the Skylab 4 mission, local areas of high humidity cannot be entirely eliminated. The reasons for the large increase in fungal isolations on Skylab 4 have been well established. Early in the Skylab 4 mission, it was discovered that mildew was present on the liquid-cooled garments which had been previously stowed aboard. A sample was taken off this growth, and one liquid cooled garment was returned for additional sampling. In general, the species of fungi isolated from surface samples and air samples were the same species isolated from the liquid cooled garment. These same microorganisms also contaminated the Petri dishes of the ED31 experiment flown on Skylab 4. It is apparent that the liquid - cooled garments were the source of spore contamination since some of these garments had not previously been removed from their original containers, but were subsequently found to harbor these microorganisms.
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This contamination was also reflected in the recovery of fungi from the crew samples collected 16 days before return from Skylab. For Skylab 2 and Skylab 3, a total of two and zero filamentous fungi, respectively, was isolated from the crew inflight. On Skylab 4 a total of 11 fungi were isolated, including a significant contamination to the astronauts. It is important to note that this contamination to the crew was demonstrated 62 days after the first exposure to the liquid-cooled garments, indicating either continued contamination from inanimate sources, abnormally slow return to normal levels, or both. The number of fungal species isolated from the 15 Command Module sites before and after each Skylab mission is shown in figure 5.These data level of fungal contamination of the crew postflight.
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Postflight Variation in the Major Components of the Autoflora Aerobic Bacteria Prior to the Skylab missions, several authors had theorized that major microflora changes might occur during space flight and that these changes might not be compatible with manâ&#x20AC;&#x2122;s health and welfare on extended missions . The theoretical change which was most often proposed called for a microbial simplification which may be defined as a major decrease in the number of different types of microorganisms in the autoflora. To evaluate this hypothesis, the variations of the aerobic bacterial portion of the total autoflora within sample collection sites were analyzed as shown in figure 6. This analysis shows that the frequency with which recovery day values lie outside the preflight range is similar for the 10 - day Apollo 14 mission and the three Skylab missions. More specifically, the total number of viable cells recovered was frequently higher postflight whereas the number of genera and species decreased in all missions except Skylab 4. Therefore, it is possible to make the following observations concerning recovery of aerobic bacteria following these space flights. Values obtained from immediate postflight sample analyses are frequently outside of the established preflight range. When different, these values most often reflect an increase in total number of viable cells and a decrease in the number of different genera and species recovered.
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Anaerobic Bacteria The anaerobic portion of the autoflora behaves quite differently than the aerobic portion. The frequency and direction of postflight change is different from each Skylab mission, but apparently is not related to mission duration (as the 10-day Apollo 14 and the 84-day Skylab 4 results are most similar). Following the Apollo 14, Skylab 2 and Skylab 4 missions, fewer viable anaerobic cells and fewer genera and species were recovered from up to 70 percent of the sites sampled. However, this is not a universal event as all of these values increased in some sample areas following Skylab 3 mission. These postflight increases were due to an unusually high level of contamination with Propionibacterium acnes on the skin of the Skylab 3 astronauts which matched exactly the increased contamination of Skylab surfaces mentioned earlier.
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Yeasts and Filamentous Fungi For the Apollo missions there was, typically, a significant reduction in the number of isolated fungal species up to the launch day. This was taken to be indicative of severely restricting opportunities of contamination to the crew for 3 weeks before flight. Analysis of postflight Apollo data indicated that exposure to the space flight environment for up to 2 weeks resulted in an even greater reduction with a relative increase in incidence of the potential pathogen, Candida albicans. Essentially the same pattern may be demonstrated from the Skylab 2 and Skylab 3 data, as shown in figure 9. However, fungal recovery was not depressed following the 59-day mission of Skylab 3, indicating increased exposure to fungi within the Skylab. Results of the same analyses for Skylab 4 are also shown in figure 9 where essentially the same pattern is again demonstrated. This is an important observation in light of the previously mentioned in-flight contamination of the Orbital Workshop and Skylab 4 crew and the fact that the Skylab 4 Pilot sustained a "rash" inflight which was presumed to be a mycotic infection and responded to treatment with Tinactin. In spite of the gross contamination, the probable mycotic infection, and the epic length of the spaceflight, approximately the same number of fungal isolates were recovered from the Skylab 4 crewmembers throughout the 17-day postflight quarantine period. This indicates that with adequate preparation, monitoring, and treatment (if necessary) it is possible to control mycological problems in space for missions of this length where the humidity is generally low.
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Intercrew Transfer of Potentially Pathogenic Microorganisms Transfer of pathogenic microorganisms between crewmembers during space flight has previously been reported for missions up to 18 days. During the Skylab series it was possible to demonstrate in-flight cross-contamination, colonization, and in-flight infection with Staphylococcus aureus. Most strains of this species, which is one of the most infectious of the common inhabitants of manâ&#x20AC;&#x2122;s autoflora, may be distinguished by their reaction with specific bacteriophages. This allows us to monitor the exchange of these microbes with greater resolution. The phage-type pattern of S. aureus recovery for Skylab 2 is shown on table 1.
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These data show that the same S. aureus phage type was repeatedly recovered from the nasal passages of the Pilot, indicating that this crewmember was a carrier of this microorganism. Although spread to the Orbital Workshop was demonstrated, there was apparently no transfer to the other crewmembers in-flight. Therefore, being restricted to a confined space for 28 days with an S. aureus carrier does not necessarily result in cross infection. A more complex situation is outlined in table 2. The data summarized in this table indicate that the Skylab 3 Commander and Pilot were both nasal carriers of S. aureus, carrying phage type 3A and 29/79, respectively. Prior to the flight, S. aureus was not recovered from any of the Scientist Pilot samples. Analyses of in-flight collected samples show that the workshop became contamined with both phage types and that type 29/79 was temporarily transferred to the Scientist Pilot. Postflight analyses show that type 3A had spread to the Pilot but, as could be expected, did not colonize this subject who was already a carrier of another phage type. Phage type 3A was repeatedly isolated from the postflight specimens of the Scientist Pilot, indicating actual colonization. This is a clear demonstration of in-flight intercrew transfer of a pathogenic species where the contaminant could be shown to have established itself as a member of the autoflora of the new host. The Pilot, a 29/79 carrier, developed a hordeolum (sty) which was successfully treated with Neosporin. The Commander, a 3A carrier, developed axillary swellings of a furuncle (boil) type which were treated with warm compresses. As neither of these infections were draining, in-flight contingency samples were not taken, so we do not know for sure the identity of the causative agent. However, we do know that the causative agent of both of these maladies is usually S. aureus, and both of these individuals were carriers of this micro-organism. Therefore, it is accurate to say that we have traced the development of a pathogenic micro-organism from its preflight carrier state in two crewmembers through in-flight contamination of the Orbital Workshop, and colonization on the third crewmember. Also, it is highly probable that this species was responsible for the active in-flight infections of the two S. aureus carriers.
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Reference:Biomedical Results from Skylab,NASA SP-377
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